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WO2025074178A1 - Procédés de préparation de colorants polymères avec des conjugués médicamenteux - Google Patents

Procédés de préparation de colorants polymères avec des conjugués médicamenteux Download PDF

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WO2025074178A1
WO2025074178A1 PCT/IB2024/058765 IB2024058765W WO2025074178A1 WO 2025074178 A1 WO2025074178 A1 WO 2025074178A1 IB 2024058765 W IB2024058765 W IB 2024058765W WO 2025074178 A1 WO2025074178 A1 WO 2025074178A1
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compound
occurrence
independently
linker
integer
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Tracy Matray
Michael VANBRUNT
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Sony Group Corp
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Sony Group Corp
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/605—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the macromolecule containing phosphorus in the main chain, e.g. poly-phosphazene
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
    • A61K49/0043—Fluorescein, used in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers

Definitions

  • Embodiments of the present disclosure are generally directed to methods for preparing polymer-drug conjugates having both fluorescent or colored moieties and biologically active moieties.
  • Targeted drug conjugates unlike, e.g., chemotherapy, deliver drugs to target cells, with little or no off-target activity.
  • targeted drug conjugates comprise a targeting molecule that is linked to a biologically active payload or drug.
  • conjugates can deliver the drug only to the intended target and minimize potential side effects.
  • Polymer-drug conjugates (PDCs) are one class of targeted drug conjugates that are of particular interest for treatments of various diseases. Polymer conjugates are found to be capable of improving the pharmacokinetic parameters of the drugs, increasing the drug stability against degradation, providing high loading capacity, and sustained release patterns, as well as avoiding premature drug release.
  • Polymer-drug conjugates integrated with an imaging agent are attractive due to the capability of providing both therapy and diagnosis or imaging functions.
  • PDCs integrated with an imaging agent helps to monitor the drug’s biodistribution and localization, which aid in a precise activation of a therapeutic response.
  • embodiments of the present disclosure are generally directed to methods for preparing polymer-drug conjugates.
  • a method for preparing a polymer-drug conjugate that permits simultaneous diagnosis and treatment options comprises reacting a compound of structure (I): with a compound of structure (II):
  • a compound of structure (I) is provided
  • the compound of structure (I) has the following structure: wherein 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 as defined herein.
  • a compound of structure (III) that is formed by reacting a compound of structure (I) with a compound of structure (II) is provided.
  • the compound of structure (III) has the following structure: wherein 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 2 , m, n, p, and w are as defined herein.
  • a compound of structure (V) that is usable as an intermediate for preparing a compound of structure (I) is provided.
  • the compound of structure (V) has the following structure: wherein 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 as defined herein.
  • a method for preparing a compound of structure (I) comprising reacting a compound of structure (V) with a compound of M 1 -G 1 , wherein M 1 -G 1 are as defined herein.
  • Amino refers to the -NH2 group.
  • Carboxy refers to the -CO2H group.
  • Cyano refers to the -CN group.
  • Alkyl refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to twelve carbon atoms (Ci-C 12 alkyl), one to eight carbon atoms (Ci-Cs alkyl) or one to six carbon atoms (Ci-Ce alkyl), and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, //-propyl,
  • alkyl groups are optionally substituted.
  • Alkylene or "alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and having from one to twelve carbon atoms, e.g., methylene, ethylene, propylene, //-butylene, ethenylene, propenylene, //-butenylene, propynylene, //-butynylene, and the like.
  • the alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, alkylene is optionally substituted.
  • alkenylene or “alkenylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond and having from two to twelve carbon atoms, e.g., ethenylene, propenylene, //-butenylene, and the like.
  • the alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond.
  • the points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, alkenylene is optionally substituted.
  • Alkynylene or “alkynylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond and having from two to twelve carbon atoms, e.g., ethynylene, propynylene, //-butynylene, and the like.
  • the alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond.
  • the points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, alkynylene is optionally substituted.
  • Alkylether refers to any alkyl group as defined above, wherein at least one carboncarbon bond is replaced with a carbon-oxygen bond.
  • the carbon-oxygen bond may be on the terminal end (as in an alkoxy group) or the carbon oxygen bond may be internal (i.e., C-O-C).
  • Alkylethers include at least one carbon oxygen bond, but may include more than one.
  • PEG polyethylene glycol
  • an alkylether group is optionally substituted.
  • Alkoxy refers to a group of the formula -ORa where R a is an alkyl group as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkoxy group is optionally substituted.
  • Heteroalkyl refers to an alkyl group, as defined above, comprising at least one heteroatom (e.g., N, O, P or S) within the alkyl group or at a terminus of the alkyl group.
  • the heteroatom is within the alkyl group (i.e., the heteroalkyl comprises at least one carbon-[heteroatom] x -carbon bond, where x is 1, 2 or 3).
  • the heteroatom is at a terminus of the alkyl group and thus serves to join the alkyl group to the remainder of the molecule (e.g., Ml-H-A), where Ml is a portion of the molecule, H is a heteroatom and A is an alkyl group).
  • heteroalkyl group is optionally substituted.
  • exemplary heteroalkyl groups include ethylene oxide (e.g., polyethylene oxide), optionally including phosphorous-oxygen bonds, such as phosphodiester bonds.
  • Heteroalkoxy refers to a group of the formula -ORa where Ra is a heteroalkyl group as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, a heteroalkoxy group is optionally substituted.
  • heteroalkylene group is optionally substituted.
  • exemplary heteroalkylene groups include ethylene oxide (e.g., polyethylene oxide) and the "C,” “HEG,” and “PEG IK” linking groups illustrated below:
  • Multimers of the above C-linker, HEG linker and/or PEG IK linker are included in various embodiments of heteroalkylene linkers.
  • n ranges from 19-25, for example n is 19, 20, 21, 22, 23, 24, or 25.
  • Multimers may comprise, for example, the following structure: wherein x is 0 or an integer greater than 0, for example, x ranges from 0-100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10).
  • the linker is a heteroatomic linker (e.g., comprising 1-10 Si, N, O, P, or S atoms), a heteroalkylene (e.g., comprising 1-10 Si, N, O, P, or S atoms and an alkylene chain) or an alkylene linker (e.g., comprising 1-12 carbon atoms).
  • a heteroalkylene linker comprises the following structure: wherein: x 9 and x 10 are each independently a integer greater than 0.
  • the heteroalkylene linker comprises at least one S-S bond.
  • physiologically cleavable linker refers to a molecular linkage that can be split or separated a prescribed manner, resulting in two or more separate molecules while in the presence of an in vivo or in vitro environment of an organism or cell system.
  • physiological conditions that induce such a cleavage or scission event may include a temperature ranging from about 20 to 40°C, an atmospheric pressure of about 1 atm (101 kPa or 14.7 psi), a pH of about 6 to 8, a glucose concentration of about 1 to 20 mM, atmospheric oxygen concentration, and earth gravity.
  • physiological conditions include enzymatic conditions (z.e., enzymatic cleavage).
  • cathepsins are proteases cleaving proteins at a low pH.
  • Cathepsins includes cathepsin A, cathepsin B, cathepsin D, cathepsin K, and cathepsin V. Bond cleavage or scission can be homolytic or heterolytic.
  • Physiological pH refers to a pH range about 4 to 8 at a temperature ranging from about 20 to 40°C, an atmospheric pressure of about 1 atm (101 kPa or 14.7 psi).
  • Heteroalkynylene is a heteroalkylene comprising at least one carbon-carbon triple bond. Unless stated otherwise specifically in the specification, a heteroalkynylene group is optionally substituted.
  • Rd is a counter ion (e.g., Na + and the like) and provided that: i) Ra is S; ii) Rb is S- or SRd; iii)Rc is SH, S- or SRd; or iv) a combination of
  • Carbocyclic refers to a stable 3- to 18-membered aromatic or non-aromatic ring comprising 3 to 18 carbon atoms. Unless stated otherwise specifically in the specification, a carbocyclic ring may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems, and may be partially or fully saturated. Non-aromatic carbocyclyl radicals include cycloalkyl, while aromatic carbocyclyl radicals include aryl. Unless stated otherwise specifically in the specification, a carbocyclic group is optionally substituted.
  • Cycloalkyl refers to a stable non-aromatic monocyclic or polycyclic carbocyclic ring, which may include fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond.
  • Monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • Polycyclic cycloalkyls include, for example, adamantyl, norbomyl, decalinyl, 7,7-dimethyl- bicyclo-[2.2.1]heptanyl, and the like. Unless stated otherwise specifically in the specification, a cycloalkyl group is optionally substituted.
  • Aryl refers to a ring system comprising at least one carbocyclic aromatic ring.
  • an aryl comprises from 6 to 18 carbon atoms.
  • the aryl ring may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems.
  • Aryls include, but are not limited to, aryls derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless stated otherwise specifically in the specification, an aryl group is optionally substituted.
  • Heterocyclic refers to a stable 3- to 18-membered aromatic or non-aromatic ring comprising one to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur.
  • the heterocyclic ring may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclic ring may be optionally oxidized; the nitrogen atom may be optionally quatemized; and the heterocyclic ring may be partially or fully saturated.
  • heteroaryls examples include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, pyrazolopyrimidinyl, quinuclidinyl, thiazolid
  • Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, benzoxazolinonyl, benzimidazolthionyl, carbazolyl, cin
  • the suffix "-ene” refers to a particular structural feature (e.g., alkyl, aryl, heteroalkyl, heteroaryl) attached to the rest of the molecule through a single bond and attached to a radical group through a single bond.
  • the suffix "-ene” refers to a linker having the structural features of the moiety to which it is attached.
  • the points of attachment of the "-ene" chain to the rest of the molecule and to the radical group can be through one atom of or any two atoms within the chain.
  • an arylene refers to a divalent linker comprising an aryl moiety as defined herein
  • a heteroarylene refers to a divalent linker comprising a heteroaryl moiety as defined herein.
  • Fused refers to a ring system comprising at least two rings, wherein the two rings share at least one common ring atom, for example two common ring atoms.
  • the fused ring is a heterocyclyl ring or a heteroaryl ring
  • the common ring atom(s) may be carbon or nitrogen.
  • Fused rings include bicyclic, tricyclic, tertracyclic, and the like.
  • substituted means any of the above groups (e.g., alkyl, alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene, heteroalkynylene, alkoxy, alkylether, phosphoalkyl, phosphoalkylether, thiophosphoalkyl, thiophosphoalkylether, carbocyclic, cycloalkyl, aryl, heterocyclic and/or heteroaryl) wherein at least one hydrogen atom (e.g., 1, 2, 3 or all hydrogen atoms) is replaced by a bond to a non-hydrogen atoms such as, but not limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl
  • Substituted also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g, a double- or triple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles.
  • a higher-order bond e.g, a double- or triple-bond
  • nitrogen in groups such as imines, oximes, hydrazones, and nitriles.
  • R g and Rh are the same or different and independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, A-heterocyclyl, heterocyclylalkyl, heteroaryl, A-heteroaryl and/or heteroarylalkyl.
  • Substituted further means any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, //-heterocyclyl, heterocyclylalkyl, heteroaryl, /V-heteroaryl and/or heteroarylalkyl group.
  • each of the foregoing substituents may also be optionally substituted with one or more of the above substituents.
  • Conjugation refers to the overlap of one p-orbital with another p-orbital across an intervening sigma bond. Conjugation may occur in cyclic or acyclic compounds.
  • a “degree of conjugation” refers to the overlap of at least one p-orbital with another p-orbital across an intervening sigma bond. For example, 1, 3-butadine has one degree of conjugation, while benzene and other aromatic compounds typically have multiple degrees of conjugation. Fluorescent and colored compounds typically comprise at least one degree of conjugation.
  • Fluorescent refers to a molecule which is capable of absorbing light of a particular frequency and emitting light of a different frequency. Fluorescence is well-known to those of ordinary skill in the art.
  • Cold refers to a molecule which absorbs light within the colored spectrum (i.e., red, yellow, blue and the like).
  • 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, without limitation, RNA, DNA, oligonucleotides, modified or derivatized nucleotides, enzymes, receptors, prions, receptor ligands (including hormones), antibodies, antigens, and toxins, as well as bacteria, viruses, blood cells, and tissue cells.
  • the visually detectable biomolecules of the disclosure are prepared, as further described herein, by contacting a biomolecule with a compound having a reactive group that enables attachment of the biomolecule to the compound via any available atom or functional group, such as an amino, hydroxy, carboxyl, or sulfhydryl group on the biomolecule.
  • a “reactive group” is a moiety capable of reacting with a second reactive groups (e.g., a “complementary reactive group”) to form one or more covalent bonds, for example by a displacement, oxidation, reduction, addition or cycloaddition reaction.
  • Exemplary reactive groups are provided in Table 1, and include for example, nucleophiles, electrophiles, dienes, dienophiles, aldehyde, oxime, hydrazone, alkyne, amine, azide, acylazide, acylhalide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester, ketone, a,P-unsaturated carbonyl, alkene, maleimide, a-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin, thiirane and the like.
  • Bio-conjugation or “bio-conjugate” and related variations refer to a chemical reaction strategy for forming a stable covalent bond between two molecules.
  • bio-conjugation is generally used when one of the molecules is a biomolecule (e.g., an antibody), but can be used to describe forming a covalent bond with a non-biomolecule (e.g., a polymeric resin).
  • a non-biomolecule e.g., a polymeric resin.
  • the product or compound resulting from such a reaction strategy is a "conjugate,” “bio-conjugate” or a grammatical equivalent.
  • visible and “visually detectable” are used herein to refer to substances that are observable by visual inspection, without prior illumination, or chemical or enzymatic activation. Such visually detectable substances absorb and emit light in a region of the spectrum ranging from about 300 to about 900 nm. Preferably, such substances are intensely colored, preferably having a molar extinction coefficient of at least about 40,000, more preferably at least about 50,000, still more preferably at least about 60,000, yet still more preferably at least about 70,000, and most preferably at least about 80,000 M ⁇ cm’ 1 .
  • the compounds of the disclosure may be detected by observation with the naked eye, or with the aid of an optically based detection device, including, without limitation, absorption spectrophotometers, transmission light microscopes, digital cameras and scanners.
  • Visually detectable substances are not limited to those which emit and/or absorb light in the visible spectrum. Substances which emit and/or absorb light in the ultraviolet (UV) region (about 10 nm to about 400 nm), infrared (IR) region (about 700 nm to about 1 mm), and substances emitting and/or absorbing in other regions of the electromagnetic spectrum are also included with the scope of "visually detectable" substances.
  • UV ultraviolet
  • IR infrared
  • the term "photostable visible dye” refers to a chemical moiety that is visually detectable, as defined hereinabove, and is not significantly altered or decomposed upon exposure to light.
  • the photostable visible dye does not exhibit significant bleaching or decomposition after being exposed to light for at least one hour. More preferably, the visible dye is stable after exposure to light for at least 12 hours, still more preferably at least 24 hours, still yet more preferably at least one week, and most preferably at least one month.
  • Non-limiting examples of photostable visible dyes suitable for use in the compounds and methods of the disclosure include azo dyes, thioindigo dyes, quinacridone pigments, dioxazine, phthalocyanine, perinone, diketopyrrolopyrrole, quinophthalone, and truarycarbonium.
  • perylene derivative is intended to include any substituted perylene that is visually detectable. However, the term is not intended to include perylene itself.
  • anthracene derivative e.g., perylene, pyrene, anthracene or naphthalene derivative
  • a derivative is an imide, bisimide or hydrazamimide derivative of perylene, anthracene, naphthalene, or pyrene.
  • the visually detectable molecules of various embodiments of the disclosure are useful for a wide variety of analytical applications, such as biochemical and biomedical applications, in which there is a need to determine the presence, location, or quantity of a particular analyte (e.g., biomolecule).
  • the disclosure provides a method for visually detecting a biomolecule, comprising: (a) providing a biological system with a visually detectable biomolecule comprising the compound of structure (III) linked to a biomolecule; and (b) detecting the biomolecule by its visible properties.
  • the phrase "detecting the biomolecule by its visible properties” means that the biomolecule, without illumination or chemical or enzymatic activation, is observed with the naked eye, or with the aid of a optically based detection device, including, without limitation, absorption spectrophotometers, transmission light microscopes, digital cameras and scanners.
  • a densitometer may be used to quantify the amount of visually detectable biomolecule present.
  • the relative quantity of the biomolecule in two samples can be determined by measuring relative optical density. If the stoichiometry of dye molecules per biomolecule is known, and the extinction coefficient of the dye molecule is known, then the absolute concentration of the biomolecule can also be determined from a measurement of optical density.
  • biological system is used to refer to any solution or mixture comprising one or more biomolecules in addition to the visually detectable biomolecule.
  • biological systems include cells, cell extracts, tissue samples, electrophoretic gels, assay mixtures, and hybridization reaction mixtures.
  • Antineoplastic or “antineoplastic agents” refer to any molecules blocking the formation of neoplasms where the growth of the neoplasms may become cancer.
  • Antineoplastic agents may include azacitidine, capecitabine, carmofur, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, nelarabine, pentostatin, tegafur, tioguanine, methotrexate, pemetrexed, raltitrexed, hydroxycarbamide, irinotecan, topotecan, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, valrubicin, etoposide, teniposide, cabazitaxel, docetaxel, paclitaxel, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, bendamustine, busulfan, carmustine, chlorambucil, chlormethine,
  • Self- immolative group refers to a group that undergoes an electronic cascade which results in the release of the group to which it is attached.
  • the self- immolative group comprises one or more groups which 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.
  • the self-immolative group comprises valine (Vai) and citrulline (Cit) or valine (Vai) and alanine (Ala).
  • Spacer group refers to a linker covalently bonding between the self-immolative group and an antineoplastic agent.
  • the spacer group is para-aminobenzylcarbamate (PBA).
  • a “solid support residue” refers to the functional group remaining attached to a molecule when the molecule is cleaved from the solid support. Solid support residues are known in the art and can be easily derived based on the structure of the solid support and the group linking the molecule thereto.
  • a “targeting moiety” is a moiety that selectively binds or associates with a particular target, such as an analyte molecule. "Selectively" binding or associating means a targeting moiety preferentially associates or binds with the desired target relative to other targets.
  • the compounds disclosed herein include linkages to targeting moieties for the purpose of selectively binding or associating the compound with an analyte of interest (i.e., the target of the targeting moiety), thus allowing detection of the analyte.
  • exemplary targeting moieties include, but are not limited to, antibodies, antigens, nucleic acid sequences, enzymes, proteins, cell surface receptor antagonists, and the like.
  • the targeting moiety is a moiety, such as an antibody, that selectively binds or associates with a target feature on or in a cell, for example a target feature on a cell membrane or other cellular structure, thus allowing for detection of cells of interest.
  • Target moieties Small molecules that selectively bind or associate with a desired analyte are also contemplated as targeting moieties in certain embodiments.
  • targeting moieties include small molecules that selectively bind or associate with a desired analyte.
  • One of skill in the art will understand other analytes, and the corresponding targeting moiety, that will be useful in various embodiments.
  • 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 natural and unnatural bases.
  • Non-limiting examples of base pairing moieties are RNA and DNA bases such adenosine, guanosine, thymidine, cytosine and uridine and analogues thereof.
  • Embodiments of the disclosure disclosed herein are also meant to encompass all compounds being isotopically-labelled by having one or more atoms replaced by an atom having a different atomic mass or mass number.
  • isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as 2 H, 3 H, U C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 0, 31 P, 32 P, 35 S, 18 F, 35 C1, 123 I, and 125 I, respectively.
  • Isotopically-labeled compounds of structure (III) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described below and in the following Examples using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.
  • “Stable compound” and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • “Optional” or “optionally” means that the subsequently described event or circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not.
  • “optionally substituted alkyl” means that the alkyl group may or may not be substituted and that the description includes both substituted alkyl groups and alkyl groups having no substitution.
  • Salt includes both acid and base addition salts.
  • Acid addition salt refers to those salts which are formed with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4- acetamidobenzoic acid, camphoric acid, camphor- 10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid
  • Base addition salt refers to those salts which are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like.
  • Particularly preferred organic bases are isoprop
  • Crystallizations may produce a solvate of the compounds described herein.
  • Embodiments of the present disclosure include all solvates of the described compounds.
  • the term "solvate" refers to an aggregate that comprises one or more molecules of a compound of the disclosure with one or more molecules of solvent.
  • the solvent may be water, in which case the solvate may be a hydrate.
  • the solvent may be an organic solvent.
  • the compounds of the present disclosure may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms.
  • the compounds of the disclosure may be true solvates, while in other cases the compounds of the disclosure may merely retain adventitious water or another solvent or be a mixture of water plus some adventitious solvent.
  • Embodiments of the compounds of the disclosure may contain one or more stereocenters and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids.
  • Embodiments of the present disclosure are meant to include all such possible isomers, as well as their racemic and optically pure forms.
  • Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization.
  • Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC).
  • HPLC high pressure liquid chromatography
  • a “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable.
  • the present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.
  • a “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule.
  • the present disclosure includes tautomers of any said compounds. Various tautomeric forms of the compounds are easily derivable by those of ordinary skill in the art.
  • Methods for preparation of compounds useful as covalent linkers between biologically active moieties such as antineoplastic agents or enediyne antitumor antibiotics and targeting moieties are provided.
  • Methods of the present disclosure allow attaching amine-containing biologically active moieties to the polymer backbone via physiologically cleavable or non- cleavable linkers.
  • the procedures described in the present disclosure provide the ability to selectively install the physiologically cleavable and/or non-cleavable linkers. This allows for synthesizing a compound which has both physiologically cleavable and non-cleavable linkers with one or more biologically active moieties.
  • the biologically active moieties can be cleaved sequentially depending on physiological conditions.
  • a compound with both multiple biologically active moieties and fluorescent or colored moieties attached by physiologically cleavable and/or non-cleavable linkers can be synthesized.
  • Methods of the present disclosure also allow for control of the number and type of biologically active moieties that are attached to the polymer backbone and any subsequent targeting moiety, spacing between the neighboring biologically active moieties on the polymer backbone (e.g., how far or close the biologically active moieties), and spacing between the polymer backbone and the biological active moiety (e.g., the length of linkers of the polymer backbone).
  • the resulting compounds includes combinations of therapeutic agents, targeting moieties, and dye moieties (e.g., chromophores or fluorophores) that can be used for simultaneous targeting, treatment, and detection.
  • dye moieties e.g., chromophores or fluorophores
  • the ease of coupling polymer-drug constructs to targeting agents such as antibodies, antibody fragments, proteins or other clinically interesting agents provides utility to a wide variety of interesting applications (e.g., surface chemistries, assay development, etc.).
  • the compounds formed by methods of the present disclosure also provide other desirable properties, including enhanced permeability and retention effects.
  • the chemical features of embodiments of the present compounds can be adjusted to modulate the compound’s ability to permeate diseased cells/tissues and be retained within the same. These features allow effective delivery of biologically active agents by increasing permeation and increasing efficacy by enhancing retention.
  • a method for preparing polymer-drug conjugates containing both biologically active moieties and fluorescent or colored dyes comprising reacting a compound of structure (I): with a compound of structure (II):
  • M 1 is, at each occurrence, independently a moiety comprising a fluorescent or colored dye
  • M 2 is a moiety comprising a biologically active moiety or a fluorescent or colored dye
  • G is, at each occurrence, independently a moiety comprising a reactive group, a group convertible into a reactive group, or protected analogue thereof, capable of forming a covalent bond with an amine group;
  • L 1 and L 7 are at each occurrence, independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, arylene or heteroarylene linker or combinations thereof;
  • L 2 , L 3 , L 5 , and L 5 are, at each occurrence, independently absent or an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene or heteroalkynylene linker;
  • L 4 is, at each occurrence, independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker;
  • R 3 is, at each occurrence, independently H, alkyl or alkoxy
  • R 4 is, at each occurrence, independently OH, SH, O', S', ORd or SRa;
  • R 5 is, at each occurrence, independently oxo, thioxo or absent;
  • R 6 and R 7 are, at each occurrence, independently H, OH, or halo;
  • Ra is O or S
  • Rb is OH, SH, O', S', ORa or SRa;
  • Re is OH, SH, O', S', ORa, OL', SRd, alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkylether, alkoxyalkylether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether;
  • Rd is a counter ion
  • Q is, at each occurrence, independently a moiety comprising a reactive group, or protected form thereof, capable of forming a covalent bond with a complementary reactive group Q' on a targeting moiety;
  • L' is, at each occurrence, independently a linker comprising a covalent bond to Q, a targeting moiety, a linker comprising a covalent bond to a targeting moiety, a linker comprising a covalent bond to a solid support, a linker comprising a covalent bond to a solid support residue, a solid support residue, a linker comprising a covalent bond to a nucleoside, or a linker comprising a covalent bond to a further compound of structure (I); m and p are, at each occurrence, independently an integer of zero or greater; w is, at each occurrence, an integer of one or greater; and n is an integer of one or greater, thereby: i) forming a bond between G of the compound of structure (I) and -NHR 8 of the compound of structure (II).
  • the various linkers and substituents e.g., M 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 , and Q
  • the optional substituent is selected to optimize the water solubility or other property of the compound of structure (I).
  • each alkyl, alkoxy, alkylether , alkoxyalkylether, phosphoalkyl, thiophosphoalkyl, phosphoalkylether and thiophosphoalkylether in the compound of structure (I) is optionally substituted with one more substituent selected from the group consisting of hydroxyl, alkoxy, alkylether , alkoxyalkylether, sulfhydryl, amino, alkylamino, carboxyl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether and thiophosphoalkylether.
  • the compound of structure (I) has the following structure (IA) or (IB):
  • the linker L 1 can be used as a point of attachment of the G group to the reminder of the compound.
  • L 1 comprises a linker that is non- cleavable under physiological conditions. Accordingly, in some embodiments, L 1 comprises an amide bond, an ester bond, a disulfide bond, a hydrazone, a phosphotriester, a diester, P- glucuronide, a double bond, a triple bond, an ether bond, a ketone, a diol, or combinations thereof.
  • L 1 comprises tert-butyloxycarbonyl, paramethoxybenzyl, dialkyl or diaryldialkoxysilane, orthoester, acetal, P-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vivinal diol, pyrophosphate diester, or valine citrulline.
  • L 1 is, at each occurrence, independently a linker that is cleavable at a pH ranging from 6 to 8.
  • L 1 is a linker that is cleavable at pH 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0.
  • L 1 is, at each occurrence, independently a linker that is cleavable at a temperature ranging from 20°C to 40°C, from 25°C to 35°C, from 30°C to 35°C, from 30°C to 37°C, from 35°C to 37°C, from 35°C to 40°C, from 32°C to 38°C.
  • L is, at each occurrence, independently a linker that is cleavable at a temperature of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C.
  • L 1 is, at each occurrence, independently a linker that is cleavable by an enzyme.
  • the enzyme is a hydrolase, an oxidoreductase or a lyase.
  • 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, EC4.4, EC 4.5, EC 4.6, or EC 4.99 enzyme.
  • L ⁇ G has the following structure:
  • L la is, at each occurrence, unsubstituted. In some more specific embodiments, L la is, at each occurrence independently a pyrimidine. In some more specific embodiments, L la is, at each occurrence, independently cytosine or thymine. In some embodiments, L la is, at each occurrence, independently selected from cytosine and thymine such that the compound comprises a sequence of cytosine and thymine bases capable of triplex formation with a target DNA sequence.
  • L la has one of the following structures:
  • L lb is, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene, heteroalkynylene, alkyl eneheteroary 1 enealkyl ene, alky 1 eneheterocy cly 1 enealky 1 ene, alkyl enecarb ocy cly 1 enealky 1 ene, heteroalkyl eneheteroary 1 enealky 1 ene, heteroalkyleneheterocyclylenealkylene, heteroalkylenecarbocyclylenealkylene, heteroalkyleneheteroaryleneheteroalkylene, heteroalkyleneheterocyclyleneheteroalkylene, heteroalkylenecarbocyclyleneheteroaryleneheteroalkylene, heteroalkyleneheterocyclyleneheteroalkylene, heteroalkylenecarbocyclylene
  • L a and L b together (/.e., has one of the following structures:
  • at least one occurrence of L 1 comprises one or more amino acid residues.
  • the amino acid residue is valine.
  • at least one occurrence of L 1 comprises one of the following structures:
  • * indicates a bond to the adjacent phosphorous atom.
  • z is an integer from 3 to 8. In some embodiments, z is 3. In some embodiments, z is 4. In some embodiments, z is 5. In some embodiments, z is 6. In some embodiments, z is 7. In some embodiments, z is 8.
  • 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.
  • z is an integer from 3 to 8. In some other embodiments, z is an integer from 22 to 26. In some other embodiments, z ranges from 19 to 28. In some embodiments, at each occurrence, in compounds of structure (I) comprises one of the following structures:
  • p wing structure in some other specific embodiments, comprises the following structure:
  • L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently absent (i.e., a direct bond) or independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker. In some embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently alkylene. In more embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently Ci-Ce alkylene, C2-C6 alkenylene or C2-C6 alkynylene.
  • the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Cr> alkylene linker.
  • at least one occurrence of L 2 , L 3 , L 5 , and L s are a direct bond.
  • at least one occurrence of L 2 is absent.
  • L 2 is absent at each occurrence.
  • the compound of structure (I) has the following structure (IE) or (IF): wherein y 1 , y 2 , y 3 , and y 4 are, at each occurrence, independently an integer from 0 to 6.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 0 (/'. ⁇ ?., each of L 2 , L 3 , L 5 , and L 6 is a direct bond). In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 1. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 2. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 3.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 4. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 5. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 6. In some more specific embodiments, y 1 , y 2 , y 3 , and y 4 are each 0 or 1 at each occurrence.
  • the linker L 7 can be used as a point of attachment of the M 1 moieties to the remainder of the compound (e.g., heteroalkylene backbone).
  • a synthetic precursor to the compound of structure (I) is prepared, and the M 1 moieties are attached to the synthetic precursor using any number of facile methods known in the art, for example methods referred to as "click chemistry.”
  • click chemistry any reaction which is rapid and substantially irreversible can be used to attach M 1 to the synthetic precursor to form a compound of structure (I).
  • Exemplary reactions include the copper catalyzed reaction of an azide and alkyne to form a triazole (Huisgen 1, 3-dipolar cycloaddition), reaction of a diene and dienophile (Diels-Alder), strain-promoted alkyne-nitrone cycloaddition, strain-promoted cycloalkyne-azide cycloaddition (Cu-free click), reaction of a strained alkene with an azide, tetrazine or tetrazole, alkene and azide [3+2] cycloaddition, alkene and tetrazine inverse-demand Diels-Alder, alkene and tetrazole photoreaction and various displacement reactions, such as displacement of a leaving group by nucleophilic attack on an electrophilic atom.
  • a triazole Huisgen 1, 3-dipolar cycloaddition
  • Diels-Alder Die
  • L 7 is at each occurrence, independently a linker comprising a functional group capable of formation by reaction of two complementary reactive groups, for example a functional group which is the product of one of the foregoing "click" reactions.
  • the functional group can be formed by reaction of an oxime, hydrazone, alkyne, amine, azide, acylazide, acylhalide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester (e.g., N- hydroxysuccinimide ester), ketone, a,[3-unsaturated carbonyl, alkene, maleimide, a-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin or
  • the functional group can be formed by reaction of an alkyne and an azide. In other embodiments, for at least one occurrence of L 7 , the functional group can be formed by reaction of an amine (e.g., primary amine) and an N-hydroxysuccinimide ester or isothiocyanate.
  • an amine e.g., primary amine
  • the functional group comprises an alkene, ester, amide, thioester, disulfide, carbocyclic, heterocyclic or heteroaryl group. In more embodiments, for at least one occurrence of L 7 , the functional group comprises an alkene, ester, amide, thioester, thiourea, disulfide, carbocyclic, heterocyclic or heteroaryl group. In other embodiments, the functional group comprises an amide or thiourea. In some more specific embodiments, for at least one occurrence of L 7 , L 7 is a linker comprising a triazolyl functional group. In some related embodiments, L 7 , at each occurrence, independently comprises a triazolyl functional group. While in other embodiments, for at least one occurrence of L 7 is a linker comprising an amide or thiourea functional group.
  • At least one occurrence of L 7 comprises one of the following structures:
  • the linker L 7 has the following structure:
  • I7-M 1 has one of the following structures: wherein L 7a and L 7b are each independently optional linkers.
  • L 7a or L 7b , or both is absent. In other embodiments, L 7a or L 7b , or both, is present.
  • L 7a and L 7b when present, are each independently alkylene or heteroalkylene.
  • L 7a and L 7b when present, independently have one of the following structures:
  • M L -L 7 has one of the following structures: wherein a, b, c, and d are each independently an integer ranging from 1-6. In some embodiments, a, b, c, and d are each independently an integer 1. In some embodiments, a, b, c, and d are each independently an integer 2. In some embodiments, a, b, c, and d are each independently an integer 3. In some embodiments, a, b, c, and d are each independently an integer 4. In some embodiments, a, b, c, and d are each independently an integer 5. In some embodiments, a, b, c, and d are each independently an integer 6.
  • At least one occurrence of L 7 comprises one of the following structures: In some embodiments, at least one occurrence of L 7 comprises one of the following structures:
  • At least one occurrence of R 3 is H. In some embodiments, at least one occurrence of R 1 is alkyl. In some embodiments, at least one occurrence of R 1 is alkoxy. In certain embodiments, each occurrence of R 1 is H.
  • At least one occurrence of R 4 is OH. In some embodiments, at least one occurrence of R 4 is SH. In some embodiments, at least one occurrence of R 4 is O'. In some embodiments, at least one occurrence of R 4 is S'. In certain embodiments, each occurrence of R 4 is O'.
  • Q a targeting moiety
  • an analyte e.g., analyte molecule
  • solid support e.g., alyte molecule
  • solid support residue e.g., a nucleoside or a further compound of structure (I).
  • the linker L' can be any linker suitable for attaching Q, a targeting moiety, an analyte (e.g., analyte molecule), a solid support, a solid support residue, a nucleoside or a further compound of structure (I) to the compound of structure (I).
  • analyte e.g., analyte molecule
  • a solid support e.g., a solid support residue
  • nucleoside or a further compound of structure (I) e.g., analyte molecule
  • an analyte e.g., analyte molecule
  • solid support e.g., a solid support residue
  • nucleoside or a further compound of structure (I) e.g., analyte molecule
  • nucleoside e.g., a further compound of structure (I)
  • nucleoside e.g., a further compound of structure (I)
  • L' has the following structure: wherein: m" and n" are independently an integer from 1 to 10;
  • R e is H, an electron pair or a counter ion
  • L" is R e or a direct bond or linkage to: Q, a targeting moiety, an analyte (e.g., analyte molecule), a solid support, a solid support residue, a nucleoside or a further compound of structure (I).
  • analyte e.g., analyte molecule
  • solid support e.g., a solid support residue
  • nucleoside e.g., nucleoside or a further compound of structure (I).
  • n" is an integer from 4 to 10, for example 4, 6 or 10. In other embodiments n" is an integer from 3 to 6, for example 3, 4, 5 or 6. In some embodiments, n" is an integer from 18-28, for example, from 21-23.
  • L" is an alkylene, alkyleneheterocyclylene, alkyleneheterocyclylenealkylene, alkylenecyclylene, alkylenecyclylenealkylene, heteroalkylene, heteroalkyleneheterocyclylene, heteroalkyl eneheterocyclyleneheteroalkylene, heteroalkylenecyclylene, or heteroalkylenecycleneheteroalkylene moiety.
  • L" comprises an alkylene oxide, phosphodiester moiety, sulfhydryl, disulfide or maleimide moiety or combinations thereof.
  • the targeting moiety is an antibody or cell surface receptor antagonist.
  • the antibody or cell surface receptor antagonist is an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a folate, or a MET inhibitor.
  • the targeting moiety is a monoclonal antibody.
  • the monoclonal antibody is Abciximab, Adalimumab, Alemtuzumab, Alirocumab, Avibactam, Basiliximab, Benralizumab, Bezlotoxumab, Blinatumomab, Brodalumab, Burosumab, Canakinumab, Caplacizumab, Certolizumab pegol, Daclizumab, Denosumab, Dupilumab, Eculizumab, Emicizumab, Erenumab, Evolocumab, Fremanezumab, Galcanezumab, Golimumab, Guselkumab, Ibalizumab, Idarucizumab, Infliximab, Itolizumab, Ixekizumab, Lanadelumab, Lokivetmab, Mepolizumab, Natalizumab, Obiltoxaximab
  • the analyte molecule is a nucleic acid, amino acid or a polymer thereof. In other embodiments, the analyte molecule is an enzyme, receptor, receptor ligand, antibody, glycoprotein, aptamer or prion.
  • the solid support is a polymeric bead or nonpolymeric bead.
  • R 1 or R 2 has one of the following structures:
  • R 1 or R 2 has the following structure:
  • the analyte molecule is a nucleic acid, amino acid or a polymer thereof In some embodiments, the analyte molecule is an enzyme, receptor, receptor ligand, antibody, glycoprotein, aptamer or prion. In some embodiments, the targeting moiety is an antibody or cell surface receptor antagonist. In further some embodiments, the solid support is a polymeric bead or non-polymeric bead.
  • Q is, at each occurrence, independently a moiety comprising a reactive group capable of forming a covalent bond with an analyte molecule or a solid support.
  • Q is, at each occurrence, independently a moiety comprising a reactive group capable of forming a covalent bond with a complementary reactive group Q'.
  • Q' is present on a further compound of structure (I) (e.g., in the R 2 or R 3 position), and Q and Q' comprise complementary reactive groups such that reaction of the compound of structure (I) and the further compound of structure (I) results in covalently bound dimer of the compound of structure (I).
  • Multimer compounds of structure (I) can also be prepared in an analogous manner and are included within the scope of embodiments of the invention.
  • the type of Q group and connectivity of the Q group to the remainder of the compound of structure (I) is not limited, provided that Q comprises a moiety having appropriate reactivity for forming the desired bond.
  • Q is a moiety which is not susceptible to hydrolysis under aqueous conditions, but is sufficiently reactive to form a bond with a corresponding group on an analyte molecule or solid support (e.g., an amine, azide or alkyne).
  • analyte molecule or solid support e.g., an amine, azide or alkyne
  • Q comprises a nucleophilic reactive group, an electrophilic reactive group or a cycloaddition reactive group.
  • Q comprises a sulfhydryl, disulfide, activated ester, isothiocyanate, azide, alkyne, alkene, diene, dienophile, acid halide, sulfonyl halide, phosphine, oc-haloamide, biotin, amino or maleimide functional group.
  • the activated ester is an N-succinimide ester, imidoester or polyfluorophenyl ester.
  • the alkyne is an alkyl azide or acyl azide.
  • Q groups can be conveniently provided in protected form to increase storage stability or other desired properties, and then the protecting group removed at the appropriate time for conjugation with, for example, a targeting moiety or analyte.
  • Q groups include “protected forms” of a reactive group, including any of the reactive groups described above and in the Table 1 below.
  • a “protected form” of Q refers to a moiety having lower reactivity under predetermined reaction conditions relative to Q, but which can be converted to Q under conditions, which preferably do not degrade or react with other portions of the compound of structure (I) or (II).
  • One of skill in the art can derive appropriate protected forms of Q based on the particular Q and desired end use and storage conditions. For example, when Q is SH, a protected form of Q includes a disulfide, which can be reduced to reveal the SH moiety using commonly known techniques and reagents.
  • the SH moiety will tend to form disulfide bonds with another sulfhydryl group, for example on another compound of structure (I). Accordingly, some embodiments include compounds of structure (I), which are in the form of disulfide dimers, the disulfide bond being derived from SH Q groups.
  • the Q moiety is conveniently masked (e.g., protected) as a disulfide moiety, which can later be reduced to provide an activated Q moiety for binding to a desired analyte molecule or targeting moiety.
  • the Q moiety may be masked as a disulfide having the following structure: wherein R is an optionally substituted alkyl group.
  • Q is provided as a disulfide moiety having the following structure: where n’ is an integer from 1 to 10.
  • the at least one fluorescent or colored moiety M 1 is a fluorescent moiety.
  • M 1 at each occurrence, independently comprises two or more aryl or heteroaryl rings, or combinations thereof, for example 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.
  • M 1 at each occurrence, independently comprises six aryl or heteroaryl rings, or combinations thereof.
  • the rings are fused.
  • M 1 at each occurrence, independently comprises 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.
  • M 1 at each occurrence, independently comprises a fused-multi cyclic aryl moiety comprising at least two fused rings.
  • M 1 is, at each occurrence, independently selected from the group consisting of a dimethylaminostilbene, quinacridone, fluorophenyl-dimethyl-BODIPY, bis-fluorophenyl-BODIPY, acridine, terrylene, sexiphenyl, porphyrin, benzopyrene, (fluorophenyl-dimethyl-difluorobora-diaza-indacene)phenyl, (/i/.s-fluorophenyl-difluorobora- diaza-indacene)phenyl, quaterphenyl, bi-benzothiazole, ter-benzothiazole, bi-naphthyl, bi- anthracyl, squaraine, squarylium, 9,10-ethynylanthracene, and ter-naphthyl moiety.
  • M 1 is, at each occurrence, independently selected from the group consisting of p-terphenyl, perylene, azobenzene, phenazine, phenanthroline, acridine, thioxanthrene, chrysene, rubrene, coronene, cyanine, perylene imide, perylene amide, and derivatives thereof.
  • M 1 is, at each occurrence, independently selected from the group consisting of a coumarin dye, resorufin dye, dipyrrometheneboron difluoride dye, ruthenium bipyridyl dye, thiazole orange dye, polymethine, and N-aryl-l,8-naphthalimide dye.
  • M 1 , M 2 , or M 3 are, at each occurrence, independently selected from the group consisting of a coumarin dye, boron-dipyrromethene, rhodamine, cyanine, pyrene, perylene, perylene monoimide, 6-FAM, 5-FAM, 6-FITC, 5-FITC, and derivatives thereof.
  • M 1 at each occurrence, independently has one of the following structures:
  • M 1 at each occurrence, independently has one of the following structures:
  • At least one occurrence of L 7 M 1 has the following structure: In some more specific embodiments, each occurrence of-L 7 -M' has the following structure:
  • the fluorescence intensity or efficacy toward treating diseases can be turned by selected different values of m, n, p, and w.
  • m has the ability to control the spacing between neighboring M 1 or M 2 .
  • m is an integer of zero or greater. In certain 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.
  • the fluorescence intensity may be tuned by number of fluorescent dye moieties attached to the polymeric backbone.
  • the value for p has the ability to control the brightness of compounds.
  • p is, at each occurrence, an integer of zero or greater. In some more specific embodiments, p is from 0 to 10. In some embodiments, p is from 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.
  • w is an integer of one or greater. In certain 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.
  • n is an integer of one or greater. In certain 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.
  • m is 6, p is 1, w is 1, and n is 1. In some specific embodiments, m is 6, p is 1, w is 2, and n is 1.
  • M 2 is a biologically active moiety.
  • M 2 is an antineoplastic agent.
  • the antineoplastic agent M 2 is, at each occurrence, independently selected from the group consisting of paclitaxel, camptonthecin, doxorubicin, monomethyl auristatin E (MMAE), Val-Cit-PAB-MMAE, and monomethyl auristatin F.
  • M 2 has one of the following structures:
  • the antineoplastic agent M 2 is monomethyl auristatin E. In some more specific embodiments, the antineoplastic agent M 2 has the following structure:
  • M 2 is an antitumor antibiotic.
  • the antitumor antibiotic M 2 is selected from the group consisting of peplomycin sulfate, pirarubicin hydrochloride, zinostatin stimalamer, idarubicin hydrochloride, mitomycin C, bleomycin hydrochloride, doxorubicin hydrochloride, daunorubicin hydrochloride, and epirubicin hydrochloride
  • M 2 is an enediyne antitumor antibiotic.
  • the enediyne antitumor antibiotic M 2 is calicheamicin or more specifically calicheamicin yl.
  • M 2 has one of the following structures:
  • M 2 is a fluorescent or colored dye as described herein with respect to M 1 .
  • the compound of structure (I) is a compound selected from Table 2.
  • compounds of the present disclosure that are formed by reacting compounds of structure (I) with compounds of structure (II) has the following structure (III): wherein:
  • M 1 is, at each occurrence, independently a moiety comprising a fluorescent or colored dye
  • M 2 is, at each occurrence, independently a moiety comprising a biologically active moiety or a fluorescent or colored dye, provided at least one occurrence of M 2 is a biologically active moiety;
  • L 1 and L 7 are at each occurrence, independently absent or an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, arylene or heteroarylene linker or combinations thereof;
  • L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently absent or an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene or heteroalkynylene linker;
  • L 4 is, at each occurrence, independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker;
  • R 4 is, at each occurrence, independently OH, SH, O', S', ORa or SRa;
  • R 5 is, at each occurrence, independently oxo, thioxo or absent;
  • R 5 and R 7 are, at each occurrence, independently H, OH, or halo; Ra is O or S;
  • Rb is OH, SH, O', S', ORd or SRa;
  • Rc is OH, SH, O', S', ORd, OL', SRd, alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkylether, alkoxyalkylether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether;
  • Rd is a counter ion
  • Q is, at each occurrence, independently a moiety comprising a reactive group, or protected form thereof, capable of forming a covalent bond with a complementary reactive group Q' on a targeting moiety;
  • L' is, at each occurrence, independently a linker comprising a covalent bond to Q, a targeting moiety, a linker comprising a covalent bond to a targeting moiety, a linker comprising a covalent bond to a solid support, a linker comprising a covalent bond to a solid support residue, a solid support residue, a linker comprising a covalent bond to a nucleoside, or a linker comprising a covalent bond to a further compound of structure (I); m and p are, at each occurrence, independently an integer of zero or greater; w is, at each occurrence, independently an integer of one or greater; and n is an integer of one or greater.
  • 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 , L 6 , L 7 , m, n, p, and w are defined in any one of the foregoing embodiments.
  • the bioactive moiety M 2 is covalently linked to the polymer chain via L 1 .
  • M 2 is the same. In other embodiments, each M 2 is different. In still more embodiments, one or more M 2 is the same and one or more M 2 is different.
  • At least one occurrence of L 1 comprises one of the following structures:
  • At least one occurrence of L 1 comprises one of the following structures:
  • At least one occurrence of L 1 comprises one of the following structures:
  • the resulting compound of structure (III) has the following structure (IIIA) wherein R 8 and L 1 are defined in any one of the foregoing embodiments.
  • at least one occurrence of L 4 is heteroalkylene.
  • each occurrence of L 4 is heteroalkylene.
  • at least one occurrence of L 4 comprises alkylene oxide.
  • the alkylene oxide of L 4 is ethylene oxide.
  • the ethylene oxide is polyethylene oxide.
  • L 4 at each occurrence, has the following structure: wherein: z is an integer from 1 to 100; and
  • * indicates a bond to the adjacent phosphorous atom.
  • the compound of structure (III) has the following structure (IIIB):
  • L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently absent (i.e., a direct bond) or independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker. In some embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently alkylene. In more embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently Ci-Ce alkylene, C2-C6 alkenylene or C2-C6 alkynylene.
  • the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Ci alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a C2 alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a C ; alkylene linker. In some more specific embodiments, the alkylene linker of L 3 and L 5 is a C4 alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Cs alkylene linker.
  • the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Cr> alkylene linker.
  • at least one occurrence of L 2 , L 3 , L 5 , and L 6 are a direct bond.
  • at least one occurrence of L 2 is absent.
  • L 2 is absent at each occurrence.
  • the compound of structure (IIII) has the following structure (IIIC): wherein y 1 , y 2 , y 3 , and y 4 are, at each occurrence, independently an integer from 0 to 6.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 0 (z.e., each of L 2 , L 3 , L 5 , and L 6 is a direct bond). In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 1. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 2. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 3.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 4. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 5. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 6. In some more specific embodiments, y 1 , y 2 , y 3 , and y 4 are each 0 or 1 at each occurrence.
  • the compound of structure (III) is a compound selected from Table 3.
  • the compounds in Table 3 are prepared according to the procedures set forth in the Examples.
  • Reaction Schemes illustrate exemplary methods of making compounds comprising one or more biologically active moieties and one or more fluorescent or colored moi eties (e.g., compound of structure (III)) by reacting a compound of structure (I) with a compound of structure (II)).
  • Reaction Scheme I illustrates direct reaction of G with an amine-containing compound of structure (II) to form compounds of structure (III) in instances where G is a reactive group.
  • Reaction Scheme II illustrates reaction of G with an amine-containing compound of structure (II) to form compounds of structure (IIIA) in instances where G is a group convertible into a reactive group.
  • G is carboxylic acid
  • the reaction of compound of structure (IA) and the compound of structure (II) is performed in the presence of an activating agent.
  • the activating agent converts carboxylic acid into a reactive group such as an activated ester or an anhydride
  • the carboxylic acid is converted into an activated ester, and compounds of structure (I) is reacted with an activating agent to produce compounds of structure (IA’) comprising the activated ester.
  • the activating agent comprises a guanidinium reagent, a uronium reagent, preferably 2-(H-benzotriazol-l-yl)-l, 1,3,3- tetramethyluronium hexafluorophosphate (HBTU), O-(benzotriazol-l-yl)-N,N,N',N'- tetramethyluronium tetrafluoroborate (TBTU) or 2-(lH-7-azabenzotriazol-l-yl)-l, 1,3,3- tetramethyluronium hexafluorophosphate (HATU), a benzotri azole reagent, preferably 1- hydroxybenzotriazole reagent (HOBt), an immonium reagent, a carbodiimide reagent, preferably N,N'-dicyclohexyl carbodiimide (DCC) or diisopropyl carbodiimide (DIP
  • the coupling agent is HBTU.
  • the activated ester then reacts the amino group in compound of structure (II) to form a compound of structure (IIIA).
  • the bioactive moiety M 2 thus is covalently linked to the polymer chain via the reaction of the activated ester with amine.
  • R 9 a leaving group of an activated ester.
  • R 9 has one of the following structures:
  • reaction of compound of structure (I) and compound of structure (II) may be carried out in a solution comprising an organic solvent and an activating agent. In some embodiments, the reaction may be carried out under a basic condition.
  • the organic solvent may one or more selected from the group consisting of dimethylsulfoxide (DMSO), N-methylformamide (NMP), N-methylpyrrolidone, dimethylacetamide, dimethylformamide (DMF), di chloromethane, di chloroethane, and chloroform.
  • DMSO dimethylsulfoxide
  • NMP N-methylformamide
  • N-methylpyrrolidone N-methylacetamide
  • DMF dimethylformamide
  • di chloromethane di chloroethane
  • chloroethane di chloroethane
  • chloroform chloroform.
  • the solvent is DMF.
  • the base may be one or more selected from the group consisting of diisopropylethylamine (DIPEA), tri ethyl amine, piperidine, pyrrolidine, pyridine, N- methylmorpholin (NMM), and collidine.
  • DIPEA diisopropylethylamine
  • NMM N- methylmorpholin
  • the base is DIPEA.
  • the coupling of the compound of structure (I) and the compound of structure (II) is carried out in the presence of HBTU in DMF.
  • the compounds of structure (I) may be prepared by DNA synthesis methodology.
  • Monomers e.g., phosphoramidite monomers
  • Introduction of desired moieties can be accomplished during the DNA synthesis steps by including the desired moiety as a portion of the monomer (see, e.g., L and M of the Representative DNA Synthesis Cycle, below).
  • An exemplary DNA synthesis scheme is shown below.
  • Oligomerization is initiated, typically, through the removal of a protecting group (e.g. a dimethoxytrityl group, DMTr) to reveal a free -OH (hydroxyl) group (Step 1, DETRIT YLATION).
  • a phosphoramidite monomer is introduced that reacts with the free OH group making a new covalent bond to phosphorus, with concomitant loss of the diisopropyl amine group (Step 2, COUPLING).
  • the resultant, phosphite triester is oxidized (e.g.
  • the new product, phosphate oligomer contains a DMTr protected OH group that can be deprotected to reinitiate the synthetic cycle so another phosphoramidite monomer can be appended to the oligomer.
  • Customization occurs at step 2 through the choice of phosphoramidite monomer.
  • L i.e., a linker group
  • M i.e., a fluorescent or colored dye
  • L i.e., a linker group
  • M i.e., a fluorescent or colored dye
  • L i.e., a linker group
  • M i.e., a fluorescent or colored dye
  • M can be optionally absent to incorporate desired spacing between M moieties.
  • a person of ordinary skill in the art can select multiple monomer types to arrive at compounds of the disclosure containing multiple therapeutic agents and/or other moieties (e.g., fluorophores or chromophores) with concurrent variability in linker groups.
  • R 4 " is alkoxy, haloalkyl, alkyl, an optionally substituted aryl or an optionally substituted aralkyl.
  • R 1 " is H.
  • R 1 " is trityl.
  • R 1 " is 4-methoxytrityl.
  • R 1 " is 4,4'- dimethoxytrityl.
  • R 2 " is H.
  • R 2 " is an activated phosphorus moiety.
  • R 2 " comprises the following structure: wherein:
  • R 5 " is H or cyano alkyl
  • R 6 " is, at each occurrence, independently Ci-Ce alkyl.
  • R 5 " is H. In other embodiments, R 5 " is 2-cyanoethyl.
  • At least one occurrence of R 6 " is isopropyl. In some embodiments, each occurrence of R 6 " is isopropyl.
  • R 2 " has the following structure:
  • R 4 " is C1-C4 haloalkyl. In more specific embodiments, R 4 " is -CFs. In some embodiments, R 4 " is C1-C4 alkoxy. In more specific embodiments, R 4 " is methoxy.
  • a compound of structure (IV) has the following structure:
  • compounds of structure (I) may be prepared by first synthesizing compounds of structure (V) and then coupling the fluorescent or colored dyes M 1 and biologically active moieties M 2 to the compounds of structure (V). Accordingly, some embodiments provide intermediate compounds having structure (V): wherein:
  • G is, at each occurrence, independently a moiety comprising a reactive group, a group convertible into a reactive group, or protected analogue thereof, capable of forming a covalent bond with an amine group;
  • L 1 and L 7 are at each occurrence, independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, arylene or heteroarylene linker or combinations thereof;
  • L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently absent or an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene or heteroalkynylene linker;
  • L 4 is, at each occurrence, independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker;
  • R 3 is, at each occurrence, independently H, alkyl or alkoxy
  • R 4 is, at each occurrence, independently OH, SH, O', S', ORd or SRd;
  • R 5 is, at each occurrence, independently oxo, thioxo or absent;
  • R 5 and R 7 are, at each occurrence, independently H, OH, or halo;
  • Ra is O or S
  • Rb is OH, SH, O', S', ORd or SRd;
  • Re is OH, SH, O', S', ORd, OL', SRd, alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkylether, alkoxyalkylether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether;
  • Rd is a counter ion
  • Q is, at each occurrence, independently a moiety comprising a reactive group, or protected form thereof, capable of forming a covalent bond with a complementary reactive group Q' on a targeting moiety;
  • L' is, at each occurrence, independently a linker comprising a covalent bond to Q, a targeting moiety, a linker comprising a covalent bond to a targeting moiety, a linker comprising a covalent bond to a solid support, a linker comprising a covalent bond to a solid support residue, a solid support residue, a linker comprising a covalent bond to a nucleoside, or a linker comprising a covalent bond to a further compound of Structure (I); m and p are, at each occurrence, independently an integer of zero or greater; w is, at each occurrence, an integer of one or greater; and n is an integer of one or greater.
  • the G 1 moiety in the compound of structure (V) can be selected from any moiety comprising a group having the appropriate reactivity group for forming a covalent bond with a complementary group on an M 1 moiety.
  • G 1 at each occurrence independently comprises a moiety suitable for reactions including: the copper catalyzed reaction of an azide and alkyne to form a triazole (Huisgen 1, 3-dipolar cycloaddition), reaction of a diene and dienophile (Diels-Alder), strain-promoted alkyne-nitrone cycloaddition, reaction of a strained alkene with an azide, tetrazine or tetrazole, alkene and azide [3+2] cycloaddition, alkene and tetrazine inverse-demand Diels- Alder, alkene and tetrazole photoreaction and various displacement reactions, such as displacement of a leaving group by nucleophilic
  • G 1 is, at each occurrence, independently a moiety comprising an oxime, hydrazone, alkyne, amine, azide, acylazide, acylhalide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester, ketone, a,[3-unsaturated carbonyl, alkene, maleimide, a-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin or thiirane functional group.
  • G 1 at each occurrence independently comprises an alkyne or an azide group. In other embodiments, G 1 at each occurrence, independently comprises an amino, isothiocyanate or activated ester group. In different embodiments, G 1 at each occurrence, independently comprises a reactive group capable of forming a functional group comprising an alkene, ester, amide, thioester, disulfide, carbocyclic, heterocyclic or heteroaryl group, upon reaction with the complementary reactive group. For example, in some embodiment the heteroaryl is triazolyl. In other of any of the foregoing embodiments of compound (V), G 1 is, at each —
  • G 1 at each occurrence, independently have one of the following structures:
  • At least one occurrence of G 1 is .
  • G 1 is, at each occurrence, independently-
  • At least one occurrence of G 1 is a protected form of an amine.
  • G 1 is, at a plurality of occurrences, independently a protected form of an amine.
  • G 1 is, at each occurrence, independently a protected form of an amine.
  • the protected form of the amine is a trifluoroacetate protected amine.
  • the protected form of the amine is a BOC protected amine.
  • the protected form of the amine is an Fmoc protected amine.
  • at least one occurrence of G 1 has one of the following structures:
  • G 1 at each occurrence, independently has one of the following structures:
  • L 7 -G 1 has one of the following structures:
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , L 2 , L 3 , L 4 , L 5 , or L 6 are as defined in any one of the foregoing embodiments.
  • the compound of structure (V) has the following structure (VA) or (VB):
  • the linker L 1 can be used as a point of attachment of the G group to the reminder of the compound.
  • L 1 comprises a linker that is non- cleavable under physiological conditions. Accordingly, in some embodiments, L 1 comprises an amide bond, an ester bond, a disulfide bond, a hydrazone, a phosphotriester, a diester, P- glucuronide, a double bond, a triple bond, an ether bond, a ketone, a diol, or combinations thereof.
  • L 1 comprises tert-butyloxycarbonyl, paramethoxybenzyl, dialkyl or diaryldialkoxysilane, orthoester, acetal, P-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vivinal diol, pyrophosphate diester, or valine citrulline.
  • L 1 is, at each occurrence, independently a linker that is cleavable at a pH ranging from 6 to 8.
  • L 1 is a linker that is cleavable at pH 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0.
  • L 1 is, at each occurrence, independently a linker that is cleavable at a temperature ranging from 20°C to 40°C, from 25°C to 35°C, from 30°C to 35°C, from 30°C to 37°C, from 35°C to 37°C, from 35°C to 40°C, from 32°C to 38°C.
  • L is, at each occurrence, independently a linker that is cleavable at a temperature of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C.
  • L 1 is, at each occurrence, independently a linker that is cleavable by an enzyme.
  • the enzyme is a hydrolase, an oxidoreductase or a lyase.
  • 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, EC4.4, EC 4.5, EC 4.6, or EC 4.99 enzyme.
  • L ⁇ G has the following structure: -j-L 1a -L 1 b -G
  • L la is an optionally substituted 5-9 membered heteroarylene linker.
  • L la is a substituted 5-membered heteroarylene linker.
  • L la is a substituted 6-membered heteroarylene linker.
  • L la is a substituted 7-membered heteroarylene linker.
  • L la is a substituted 8-membered heteroarylene linker.
  • L la is a substituted 9-membered heteroarylene linker.
  • L la is substituted with oxo, alkyl (e.g., methyl, ethyl, etc.) or combinations thereof.
  • L la is, at each occurrence, unsubstituted. In some more specific embodiments, L la is, at each occurrence independently a pyrimidine. In some more specific embodiments, L la is, at each occurrence, independently cytosine or thymine. In some embodiments, L la is, at each occurrence, independently selected from cytosine and thymine such that the compound comprises a sequence of cytosine and thymine bases capable of triplex formation with a target DNA sequence.
  • L lb is, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenyl ene, heteroalkynylene, alkyl eneheteroary 1 enealkyl ene, alky 1 eneheterocy cly 1 enealky 1 ene, alkyl enecarb ocy cly 1 enealky 1 ene, heteroalkyl eneheteroary 1 enealky 1 ene, heteroalkyleneheterocyclylenealkylene, heteroalkylenecarbocyclylenealkylene, heteroalkyleneheteroaryleneheteroalkylene, heteroalkyleneheterocyclyleneheteroalkylene, heteroalkylenecarbocyclyleneheteroaryleneheteroalkylene, heteroalkyleneheterocyclyleneheteroalkylene, heteroalkylenecarbocyclylene
  • L a and I? together (z.e., ? ? ) has one of the following structures:
  • At least one occurrence of L 1 comprises one or more amino acid residues.
  • the amino acid residue is valine.
  • at least one occurrence of L 1 comprises one of the following structures:
  • At least one occurrence of L 1 comprises one of the following structures: In some embodiments, at least one occurrence of L 4 is heteroalkylene. In some embodiments, each occurrence of L 4 is heteroalkylene. In some embodiments, at least one occurrence of L 4 comprises alkylene oxide. In some more specific embodiments, the alkylene oxide of L 4 is ethylene oxide. In some more specific embodiments, the ethylene oxide is polyethylene oxide. In certain embodiments, L 4 , at each occurrence, has the following structure: wherein: z is an integer from 1 to 100; and
  • the compound of structure (V) has the following structure (VC) or (VD):
  • L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently absent (z'.e., a direct bond) or independently an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker. In some embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently alkylene. In more embodiments, L 2 , L 3 , L 5 , and L 6 are, at each occurrence, independently Ci-Ce alkylene, C2-C6 alkenylene or C2-C6 alkynylene.
  • the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Ci alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a C2 alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a C3 alkylene linker. In some more specific embodiments, the alkylene linker of L 3 and L 5 is a C4 alkylene linker. In some more specific embodiments, the alkylene linker of L 2 , L 3 , L 5 , and L 6 is a Cs alkylene linker.
  • the alkylene linker of L 2 , L 3 , L 5 , and L 5 is a C& alkylene linker.
  • at least one occurrence of L 2 , L 3 , L 5 , and L 6 are a direct bond.
  • at least one occurrence of L 2 is absent.
  • L 2 is absent at each occurrence.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 0 (z.e., each of L 2 , L 3 , L 5 , and L 6 is a direct bond). In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 1. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 2. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 3.
  • y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 4. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 5. In some embodiments, y 1 , y 2 , y 3 , and y 4 are, at each occurrence, an integer 6. In some more specific embodiments, y 1 , y 2 , y 3 , and y 4 are each 0 or 1 at each occurrence.
  • the compound of structure (V) is a compound selected from Table 4.
  • Reaction Scheme III illustrates an exemplary method for preparing a compound 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 as defined above.
  • Compound of structure (V) is reacted with M ⁇ G 1 to yield compound of structure (I).
  • G 1 and G 1 represent functional groups having complementary reactivity (i.e., functional groups which react to form a covalent bond).
  • G 1 may be pendant to M 1 or a part of the structural backbone of M 1 .
  • G 1 and G 1 may be any number of functional groups described herein, such as alkyne and azide, respectively, amine and activated ester, respectively or amine and isothiocyanate, respectively, and the like.
  • Suitable protecting groups include hydroxy, amino, mercapto and carboxylic acid.
  • Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (for example, t- butyldimethylsilyl, t-butyldiphenyl silyl or trimethyl silyl), tetrahydropyranyl, benzyl, and the like.
  • Suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like.
  • Suitable protecting groups for mercapto include -C(O)-R" (where R" is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl and the like.
  • Suitable protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters.
  • Protecting groups may be added or removed in accordance with standard techniques, which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley.
  • the protecting group may also be a polymer resin such as a Wang resin, Rink resin, or a 2-chlorotrityl-chloride resin.
  • Mass spectral analysis is performed on a Waters/Micromass Quattro micro MS/MS system (in MS only mode) using MassLynx 4.1 acquisition software.
  • Mobile phase used for LC/MS on dyes is 100 mM l,l,l,3,3,3-hexafluoro-2-propanol (HFIP), 8.6 mM triethylamine (TEA), pH 8.
  • HFIP mM l,l,l,3,3,3-hexafluoro-2-propanol
  • TAA triethylamine
  • Phosphoramidites and precursor molecules are also analyzed using a Waters Acquity UHPLC system with a 2.1 mm x 50 mm Acquity BEH-Cis column held at 45 °C, employing an acetonitrile / water mobile phase gradient.
  • Compound 1-1 and Compound 1-2 were prepared by automated solid-phase synthesis (SPS) using standard phosphoramidite chemistry; analogous to the synthesis chemistry used for oligonucleotide synthesis.
  • SPS solid-phase synthesis
  • the synthesis process involves: solid phase synthesis, final deprotection and cleavage from the solid support, isolation of the compound, and analysis.
  • the general solid phase synthesis process is a repetitive cycle of four steps, with solvent washes in between each step, as follows: step 1- detrityl ati on, step 2- monomer coupling, step 3-oxidation, and step 4-capping. The synthesis cycle is repeated until all of the monomers have been sequentially incorporated into the polymer.

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Abstract

L'invention concerne un procédé de préparation de conjugués polymère-médicament contenant à la fois des fractions biologiquement actives et des colorants fluorescents ou colorés. Le procédé comprend la mise en réaction d'un composé, dont la structure (I) est la suivante : Formule (I), avec un composé biologiquement actif contenant une amine, dont la structure (II) est la suivante : M2-NHR8, où G comprend un groupe apte à former une liaison covalente avec un groupe amine, et R1, R2, R3, R4, R5, R6, R7, R8, L1, L2, L3, L 4, L 5, L6, L7, M1, m, n, p et w sont tels que définis dans la description.
PCT/IB2024/058765 2023-10-03 2024-09-09 Procédés de préparation de colorants polymères avec des conjugués médicamenteux Pending WO2025074178A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019071208A1 (fr) * 2017-10-05 2019-04-11 Sony Corporation Médicaments polymères programmables
US20190136065A1 (en) 2016-05-10 2019-05-09 Sony Corporation Compositions comprising a polymeric dye and a cyclodextrin and uses thereof
CN115260261A (zh) * 2021-04-30 2022-11-01 苏州诺维康生物科技有限公司 一种荧光染料修饰的脱氧核苷固相载体的制备方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20190136065A1 (en) 2016-05-10 2019-05-09 Sony Corporation Compositions comprising a polymeric dye and a cyclodextrin and uses thereof
WO2019071208A1 (fr) * 2017-10-05 2019-04-11 Sony Corporation Médicaments polymères programmables
CN115260261A (zh) * 2021-04-30 2022-11-01 苏州诺维康生物科技有限公司 一种荧光染料修饰的脱氧核苷固相载体的制备方法

Non-Patent Citations (2)

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Title
GREEN, T.W.P.G.M. WUTZ: "Protective Groups in Organic Synthesis", 1999, WILEY
SIGMUND HARALD ET AL: "A New Type of Fluorescence Labeling of Nucleosides, Nucleotides and Oligonucleotides", NUCLEOSIDES AND NUCLEOTIDES, vol. 16, no. 5-6, 1 May 1997 (1997-05-01), US, pages 685 - 696, XP093234419, ISSN: 0732-8311, DOI: 10.1080/07328319708002935 *

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