Classifications

• • 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
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/74—Synthetic polymeric materials
  • A61K31/80—Polymers containing hetero atoms not provided for in groups A61K31/755 - A61K31/795
• • 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
  • A61K47/68—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 antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6835—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 antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
  • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule

Definitions

• the present disclosure is generally directed to dimeric and polymeric biologically active compounds having spacing groups with or without a chromophore moiety (e.g., a fluorescent dye), compounds, and methods of treatment related to the same.
• a chromophore moiety e.g., a fluorescent dye
• 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.
• ADCs Antibody-drug conjugates
• ADCs for cancer treatment combine the targeting features of monoclonal antibodies with cancer- killing ability of cytotoxic agents to provide a therapeutic with several advantages over other chemotherapeutics.
• challenges related to the complexity of ADC constructs, specifically the chemical linker between antibody and drug has caused significant difficulties for development of new and effective therapeutics.
• Adcetris ® As of today, only Adcetris ® , Besponsa ® , Enhertu ® , Mylotarg ® , Padcev ® , Polivy ® , and Kadcyla ® are commercially available globally (Zevalin ® has been approved in China only).
• Zevalin ® has been approved in China only.
• Another embodiment provides a compound having the following structure (Ia): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , L 2 , L 3 , L 4 , L 6 , L 7 , L 9 , L 10 , L 11 , M 1 , M 2 , M 3 , l, m, n, p, and q are as defined herein.
• Another embodiment provides a compound having the following structure (Ib): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , L 3 , L 10 , M 1 , M 2 , M 3 , l, m, n, p, and q are as defined herein.
• Yet another embodiment provides a compound having the following structure (Ic): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , R 8 , R 9 , R 10 , L 3 , L 10 , M 1 , M 3 , l, m, n, p, and q are as defined herein.
• yet another embodiment provides a compound having the following structure (Id): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , R 8 , R 9 , R 10 , L 10 , M 3 , n, p, and q are as defined herein. Further, yet another embodiment provides a compound having the following structure (Ie):
• One embodiment provides a compound having the following Structure (III): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, 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 , L 8 , L 9 , L 10 , L 11 , M 1 , M 2 , M 3 , l, m, n, p, and q are as defined herein.
• Another embodiment provides a compound having the following structure (IIIa): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , L 2 , L 3 , L 4 , L 6 , L 7 , L 9 , L 10 , L 11 , M 1 , M 2 , M 3 , l, m, n, p, and q are as defined herein.
• Another embodiment provides a compound having the following structure (IIIb): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , L 3 , L 10 , M 1 , M 2 , M 3 , l, m, n, p, and q are as defined herein.
• Yet another embodiment provides a compound having the following structure (IIIc): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , R 8 , R 9 , R 10 , L 3 , L 10 , M 1 , M 3 , l, m, n, p, and q are as defined herein.
• yet another embodiment provides a compound having the following structure (IIId): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 2 , R 3 , R 6 , R 7 , R 8 , R 9 , R 10 , L 10 , M 3 , n, p, and q are as defined herein.
• FIGs.1-2 illustrate cell proliferation assay results for Her2 negative in PC3.
• DETAILED DESCRIPTION In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments of the disclosure. However, one skilled in the art will understand that embodiments of the disclosure may be practiced without these details. Unless the context requires otherwise, throughout the present specification and claims, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to”.
• Niro refers to the ⁇ NO 2 group.
• Sulfhydryl refers to the thiol 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 (C 1 -C 12 alkyl), one to eight carbon atoms (C 1 -C 8 alkyl) or one to six carbon atoms (C 1 -C 6 alkyl), and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, and the like.
• 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, n-butylene, ethenylene, propenylene, n-butenylene, propynylene, n-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.
• 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 at least one carbon-carbon double bond and having from two to twelve carbon atoms, e.g., ethenylene, propenylene, n-butenylene, and the like.
• 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., ethenylene, propenylene, n-butenylene, and the like.
• the alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond.
• the points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain.
• alkynylene is optionally substituted.
• Alkylether refers to any alkyl group as defined above, wherein at least one carbon-carbon 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. For example, polyethylene glycol (PEG) is included within the meaning of alkylether. Unless stated otherwise specifically in the specification, an alkylether group is optionally substituted.
• PEG polyethylene glycol
• Alkoxy refers to a group of the formula ⁇ OR a 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.
• Heteroalkylene refers to an alkylene group, as defined above, comprising at least one heteroatom (e.g., Si, N, O, P or S) within the alkylene chain or at a terminus of the alkylene chain.
• the heteroatom is within the alkylene chain (i.e., the heteroalkylene comprises at least one carbon-[heteroatom]x- carbon bond, where x is 1, 2 or 3).
• the heteroatom is at a terminus of the alkylene and thus serves to join the alkylene to the remainder of the molecule (e.g., M1-H-A-M2, where M1 and M2 are portions of the molecule, H is a heteroatom and A is an alkylene).
• a heteroalkylene group is optionally substituted.
• heteroalkylene groups include ethylene oxide (e.g., polyethylene oxide) and the "C” linking group and "HEG” linking groups illustrated below: Multimers of the above C linker and HEG linker are included in various embodiments of heteroalkylene linkers.
• Heteroalkenylene is a heteroalkylene, as defined above, comprising at least one carbon-carbon double bond. Unless stated otherwise specifically in the specification, a heteroalkenylene group is optionally substituted.
• 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.
• Heteroatomic in reference to a “heteroatomic linker” refers to a linker group consisting of one or more heteroatom.
• a phosphoalkyl group is optionally substituted.
• the ⁇ Oalkyl moiety in a phosphoalkyl group is optionally substituted with one or more of hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether.
• the -Oalkylether moiety in a phosphoalkylether group is optionally substituted with one or more of hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether.
• R a is O or S
• R b is OH, O-, S-, OR d or SR d
• R c is OH, SH, O-, S-, OR
• the ⁇ Oalkyl moiety in a thiophosphoalkyl group is optionally substituted with one or more of hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether.
• the -Oalkylether moiety in a thiophosphoalkyl group is optionally substituted with one or more of hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether.
• Carbocyclic refers to a stable 3- to 18-membered aromatic or non-aromatic ring comprising 3 to 18 carbon atoms.
• 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.
• 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 cyclocalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
• Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo-[2.2.1]heptanyl, and the like. Unless 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. In some embodiments, 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 quaternized; and the heterocyclic ring may be partially or fully saturated.
• heteroaryls examples include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, pyrazolopyrimidinyl, quinuclidinyl, thiazolidin
• heteroaryl refers to a 5- to 14-membered ring system comprising one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring.
• the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized.
• Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2- ⁇ ]pyridinyl, benzoxazolinonyl, benzimidazolthionyl, carbazolyl, cinnolin
• a heteroaryl group is optionally substituted.
• the suffix "-ene” refers to a particular structural feature (e.g., alkyl, aryl, heteroalkyl) attached to the rest of the molecule through a single bond and to the radical group through a single bond.
• the suffix "-ene” refers to a particular structural feature having the description given herein which is a linker between the molecule and a radical group.
• 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 alkyleneheteroalkylene refers to a linker comprising an alkylene portion and a heteroalkylene portion.
• “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. When 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.
• 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).
• a “linker” refers to a contiguous chain of at least one atom, such as carbon, oxygen, nitrogen, sulfur, phosphorous, and combinations thereof, which connects a portion of a molecule to another portion of the same molecule or to a different molecule, moiety or solid support (e.g., microparticle). Linkers may connect the molecule via a covalent bond or other means, such as ionic or hydrogen bond interactions.
• 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.
• 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 (i.e., enzymatic cleavage). Bond cleavage or scission can be homolytic or heterolytic.
• Solid support or solid resin refers to any solid substrate known in the art for solid-phase support of molecules, for example a “microparticle” refers to any of a number of small particles useful for attachment to compounds of the disclosure, including, but not limited to, glass beads, magnetic beads, polymeric beads, nonpolymeric beads, and the like.
• a microparticle comprises polystyrene beads.
• the solid support or solid resin is controlled pore glass or macroporous polystyrene.
• 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. Embodiments disclosed herein are also meant to encompass all compounds of Structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) 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, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I, respectively.
• Isotopically-labeled compounds of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) 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.
• 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, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic
• 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.
• basic ion exchange resins such
• Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, caffeine, and the like. Crystallizations may produce a solvate of the compounds described herein (e.g., a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId).
• 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.
• 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 e.g., compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) having a biomolecule linked thereto
• a biomolecule e.g., compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) having a biomolecule linked thereto
• a “reactive group” is a moiety capable of reacting with a second reactive group (e.g., a “complementary reactive group”) to form one or more covalent bonds, 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, ⁇ , ⁇ -unsaturated carbonyl, alkene, maleimide, ⁇ -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).
• 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.
• 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 -1 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.
• UV ultraviolet
• IR infrared
• other regions of the electromagnetic spectrum are also included with the scope of "visually detectable” substances.
• 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.
• 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.
• 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.
• polymer compounds 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, spatial interaction or quantity of a particular analyte (e.g., biomolecule).
• 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 the embodiments disclosed herein (e.g., compounds of structures ((I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) linked to a biomolecule; and (b) detecting the biomolecule by its visible properties.
• a biological system with a visually detectable biomolecule comprising the compound of the embodiments disclosed herein (e.g., compounds of structures ((I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) 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.
• Optional or “optionally” means that the subsequently described event or circumstances may or may not occur; such a description includes instances where the event or circumstance occurs and instances where 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.
• the charge is dependent on pH and the uncharged (e.g., protonated or salt, such as sodium or other cation) forms are also included in the scope of embodiments of the disclosure.
• Compounds As noted above, in one embodiment of the present disclosure, compounds useful as covalent linkers between biologically active moieties and targeting moieties are provided. In another embodiment, an inclusion of one or more biologically active moieties (e.g., gemcitabine) within the backbone of compounds is provided. In other embodiments, an inclusion of one or more fluorescent dyes into the compounds useful as covalent linkers between biologically active moieties and targeting moieties is provided. Further, an inclusion of one or more fluorescent dyes into the compounds allows for visualizing area that the compounds are attached.
• biologically active moieties e.g., gemcitabine
• an inclusion of one or more fluorescent dyes into the compounds useful as covalent linkers between biologically active moieties and targeting moieties is provided. Further, an inclusion of one or more fluorescent dyes into the compounds allows for visualizing area
• compounds useful as synthetic intermediates for preparation of compounds comprising one or more biologically active moieties are provided. Numerous advantages are afforded by embodiments disclosed herein, including the ability to control the number of biologically active moieties that are attached to the polymer and any subsequent targeting moiety, the ability to control the type of biologically active moieties that are attached to the polymer and any subsequent targeting moiety, the ability to control the number of fluorescent dye moieties that are attached to the polymer and any subsequent targeting moiety, and the ability to control the order of fluorescent dye moieties and biologically active moieties that are attached to the polymer and any subsequent targeting moiety.
• composition of the polymer backbone can also be selected to afford desirable solubility properties, for example, by controlling the incorporation of charged moieties (e.g., number, frequency, spacing, etc.).
• side chains can be selected to provide a source for tuning the solubility of the compounds disclosed herein.
• Monomeric units of the polymer can be selected to incorporate different anticancer therapeutics during polymer synthesis and as a post synthetic modification following polymer synthesis (e.g., coupling to an amine pendant to the polymer backbone with a therapeutic agent having an activated ester moiety).
• composition of the polymer backbone can also be selected to afford desirable fluorescent properties, for example, by controlling the incorporation of fluorescent dye moieties (e.g., number, frequency, spacing, etc.). That is, the embodiments disclosed herein also provide compounds that can advantageously include multiple therapeutic agents, for example, for complimentary or synergistic therapeutic strategies. In addition, embodiments of the present disclosure provide combinations of therapeutic agents, targeting moieties, and dye moieties (e.g., chromophores or fluorophores) that can be used for simultaneous targeting, treatment, and detection.
• fluorescent dye moieties e.g., number, frequency, spacing, etc.
• M 1 is a chromophore or fluorophore (e.g., FITC, 5-FAM, 6-FAM, and the like)
• M 2 and/or M 3 is a therapeutic agent (e.g., a drug moiety such as Auristatin F or SN 38)
• compounds disclosed herein have gemcitabine moieties embedded in the polymer backbone.
• the compounds of certain embodiments 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/tissue and be retained within the same. These features allow effective delivery of biologically active agents by increasing permeation and increasing efficacy by enhancing retention.
• At least one occurrence of L 1 , L 5 , or L 8 is alkylene. In certain embodiments, at least one occurrence of L 1 , L 5 , or L 8 is methylene. In more specific embodiments at least one occurrence of L 1 , L 5 , or L 8 is heteroalkylene. In certain embodiments, at least one occurrence of L 1 , L 5 , or L 8 comprises alkylene oxide. Further in some embodiments, the alkylene oxide is ethylene oxide. For example, the ethylene oxide is polyethylene oxide. In some embodiments, R 2 is L'. In some other embodiments, L' is a linker to a targeting moiety.
• L' is a linker to a targeting moiety, the linker comprising an alkylene oxide or phosphodiester moiety, or combinations thereof.
• L' has one of the following structures: wherein: x 1 , x 2 , x 3 , x 4 , x 5 , x 6 , x 7 and x 8 are independently an integer from 1 to 10; R b is H, an electron pair or a counter ion; L'' is the targeting moiety or a linkage to the targeting moiety.
• 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.
• EGFR epidermal growth factor receptor
• HGFR hepatocyte growth factor receptor
• IGFR insulin-like growth factor receptor
• folate a folate
• MET inhibitor a MET inhibitor
• the targeting moiety is a monoclonal antibody which includes 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, Obiltox
• R a is H or a solid support.
• R 2 has one of the following structures:
• R 3 has the following structure: .
• the compound has the following structure (Ia): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein z is, at each occurrence, independently an integer from 1 to 10.
• R 5 is, at each occurrence, independently OH, O- or OR d .
• R 4 is, at each occurrence, oxo.
• the compound has the following structure (Ib): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein x a , x b , x c , x d , x e , and x f are, at each occurrence, independently an integer from 0 to 6.
• the compound has the following structure (Ic): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 8 is, at each occurrence, independently O, NH, or NR e ; R 9 is, at each occurrence, independently H, alkyl, or optionally substituted alkyl; R 10 is, at each occurrence, independently H or F; and R e is, at each occurrence, independently alkyl or optionally substituted alkyl.
• m is an integer of zero.
• the compound has the following structure (Id): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
• the compound has the following structure (Ie):
• the compound has the following structure (IIIa): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
• the compound has the following structure (IIIb): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein x a , x b , x c , x d , x e , and x f are, at each occurrence, independently an integer from 0 to 6.
• the compound has the following structure (IIIc): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein R 8 is, at each occurrence, independently O, NH, or NR e ; R 9 is, at each occurrence, independently H, alkyl, or optionally substituted alkyl; R 10 is, at each occurrence, independently H or F; and R e is, at each occurrence, independently alkyl or optionally substituted alkyl.
• the compound has the following structure (IIId): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
• the various linkers and substituents e.g., M 1 , M 2 , M 3 , Q, R 1 , R 2 , R 3 , L 1 , L 2 , L 3 , L 4 , L 5 , L 6 , L 7 , L 8 , L 9 , L 10 , or L 11 ) in the compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) are optionally substituted with one more substituent.
• the optional substituent is selected to optimize the water solubility or other property of the compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), ((III), (IIIa), (IIIb), (IIIc), or (IIId).
• each alkyl, alkoxy, alkylether , alkoxyalkylether, phosphoalkyl, thiophosphoalkyl, phosphoalkylether and thiophosphoalkylether in the compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) are 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.
• L 3 or L 10 is, at each occurrence, independently a direct bond or an optionally substituted linker. In some embodiments, L 3 or L 10 is, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, or heteroatomic linker. In some embodiments, L 3 or L 10 is a linker comprising a functional group capable of formation by reaction of two complementary reactive groups (e.g., an azide and an alkyne).
• L 3 or L 10 is, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, alkyleneheteroarylenealkylene, alkyleneheterocyclylenealkylene, alkylenecarbocyclylenealkylene, heteroalkyleneheteroarylenealkylene, heteroalkyleneheterocyclylenealkylene, heteroalkylenecarbocyclylenealkylene, heteroalkyleneheteroaryleneheteroalkylene, heteroalkyleneheterocyclyleneheteroalkylene, heteroalkylenecarbocyclyleneheteroalkylene, alkyleneheteroaryleneheteroalkylene, alkyleneheterocyclyleneheteroalkylene, alkylenecarbocyclyleneheteroalkylene, heteroarylene, heterocyclylene, carbocyclylene, alkyleneheteroarylene, alkylene, alky
• L 3 or L 10 is optionally substituted.
• the linkers L 3 or L 10 can be used as a point of attachment of the M 1 and M 3 moieties to the remainder of the compound.
• L 3 or L 10 , or both is absent.
• L 3 or L 10 , or both is present.
• L 3 or L 10 when present, are each independently alkylene or heteroalkylene.
• at least one occurrence of L 3 or L 10 is heteroalkylene.
• at least one occurrence of L 3 or L 10 comprises oxygen.
• At least one occurrence of L 3 or L 10 has the following structure: wherein: x 9 and x 10 are each independently a integer greater than 0. In some embodiments, x 9 is 1, 2, 3, or 4. In certain embodiments, x 10 is 2, 3, 4, or 5. In some specific embodiments, x 9 is 1 or 2 and x 10 is 2, 3, or 4. In certain specific embodiments, each occurrence of L 3 or L 10 is heteroalkylene. In some more specific embodiments, each occurrence of L 3 or L 10 comprises oxygen. In certain more specific embodiments, each occurrence of L 3 or L 10 has the following structure: wherein: x 9 and x 10 are each independently a integer greater than 0. In some embodiments, x 9 is 1, 2, 3, or 4.
• x 10 is 2, 3, 4, or 5. In more specific embodiments, x 9 is 1 or 2 and x 10 is 2, 3, or 4. In certain other embodiments, at least one occurrence of L 3 or L 10 comprises the following structure: wherein: x 9 and x 10 are each independently a integer greater than 0. In certain embodiments, L 3 or L 10 further comprises a physiologically cleavable linker.
• At least one occurrence of L 3 or L 10 comprises an amide bond, an ester bond, a phosphodiester bond, a disulfide bond, a double bond, a triple bond, an ether bond, a hydrazone, an amino acid sequence comprising one or more amino acid residues, a ketone, a diol, a cyano, a nitro, or combinations thereof.
• At least one occurrence of L 3 or L 10 comprises one of the following structures:
• each occurrence of L 3 or L 10 comprises an amide bond, an ester bond, a phosphodiester bond, a disulfide bond, a double bond, a triple bond, an ether bond, a hydrazone, an amino acid sequence, a ketone, a diol, a cyano, a nitro or combinations thereof.
• each occurrence of L 3 or L 10 comprises one of the following structures:
• at least one occurrence of L 3 or L 10 has one of the following structures: .
• each occurrence of L 3 or L 10 has one of the following structures: .
• each occurrence of L 1 , L 5 , or L 9 independently comprise a phosphodiester.
• at least one occurrence of L 1 , L 5 , or L 9 comprises ethylene oxide.
• at least one occurrence of L 1 , L 5 , or L 9 comprises one of the following structures: wherein: g is an integer ranging from 1-10; and z' is an integer ranging from 1-30. In some of the foregoing embodiments, z' is 3, 6, or 11-28. In some embodiments, g ranges from 2-5. In other more specific embodiments, at least one occurrence of L 1 , L 5 , or L 9 comprises the following structure: .
• each occurrence of L 1 , L 5 , or L 9 comprises the following structure: .
• the other of R 2 or R 3 is Q or a linker comprising a covalent bond to Q.
• R c is OL′.
• L′ is a heteroalkylene linker to: Q, a targeting moiety, an analyte molecule, a solid support, a solid support residue, a nucleoside or a further compound of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), ((III), (IIIa), (IIIb), (IIIc), or (IIId).
• L′ comprises an alkylene oxide or phosphodiester moiety, or combinations thereof.
• 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 molecule, a solid support, a solid support residue, a nucleoside or a further compound of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId).
• 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 (e.g., controlled pore glass or polystyrene beads). In other embodiments, 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 (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) (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), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) and the further compound of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) results in covalently bound dimer of the compound of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId).
• Multimer compounds of structures (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) and combinations thereof can also be prepared in an analogous manner and are included within the scope of embodiments of the disclosure.
• the type of Q group and connectivity of the Q group to the remainder of the compound of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) is not particularly 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 (e.g., a biomolecule) or solid support (e.g., an amine, azide, or alkyne).
• analyte molecule e.g., a biomolecule
• solid support e.g., an amine, azide, or alkyne.
• Certain embodiments of compounds of structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), and/or (IIId) comprise Q groups commonly employed in the field of bio-conjugation.
• 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, ⁇ -haloamide, biotin, amino, or maleimide functional group.
• the activated ester is an N-succinimide ester, imidoester, or polyflourophenyl ester.
• the alkyne is an alkyl azide or acyl azide.
• Q comprises a maleimide functional group. Exemplary Q moieties are provided in Table I below.
• the other of R 2 or R 3 is a linker comprising a covalent bond to an analyte molecule or a linker comprising a covalent bond to a solid support.
• the analyte molecule is a nucleic acid or a polymer thereof or an amino acid or a polymer thereof.
• the analyte molecule is an enzyme, receptor, receptor ligand, antibody, glycoprotein, aptamer or prion.
• the solid support is a polymeric bead or non-polymeric bead.
• the targeting moiety is an antibody or cell surface receptor antagonist.
• R 2 or R 3 has one of the following structures:
• at least one occurrence of M 3 is an alkylating agent, an antimetabolite, a microtubule inhibitor, a topoisomerase inhibitor, or a cytotoxic antibiotic.
• each occurrence of M 3 is an alkylating agent, an antimetabolite, a microtubule inhibitor, a topoisomerase inhibitor, or a cytotoxic antibiotic.
• At least one occurrence of M 3 is a nitrogen mustard, a nitrosourea, a tetrazine, an aziridine, a cisplatin or cisplatin derivative, or a non-classical alkylating agent.
• At least one occurrence of M 3 is mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide, busulfan, N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, streptozotocin, dacarbazine, mitozolomide, temozolomide, thiotepa, mytomycin, diaziquone (AZQ), cisplatin, carboplatin, oxaliplatin, procarbazine, or hexamethylmelamine.
• MNU N-nitroso-N-methylurea
• BCNU carmustine
• CCNU lomustine
• Semustine MeCCNU
• fotemustine streptozotocin
• dacarbazine mitozolomide
• temozolomide temozolomide
• thiotepa myto
• At least one occurrence of M 3 is an anti-folate, a fluoropyrimidines, a deoxynucleoside analogue, or a thiopurine.
• at least one occurrence of M 3 is methotrexate, pemetrexed, fluorouracil, capecitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, pentostatin, thioguanine, and mercaptopurine.
• at least one occurrence of M 3 is an auristatin, a Vinca alkaloid, or a taxane.
• At least one occurrence of M 3 is auristatin F, auristatin E, vincristine, vinblastine, vinorelbine, vindesine, vinflunine, paclitaxel, docetaxel, etoposide, or teniposide.
• at least one occurrence of M 3 is irinotecan, SN 38, topotecan, camptothecin, doxorubicin, mitoxantrone, teniposide, novobiocin, merbarone, or aclarubicin.
• at least one occurrence of M 3 is an anthracycline or a bleomycin.
• At least one occurrence of M 3 is doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, or mitoxantrone.
• at least one occurrence of M 3 is auristatin F, monomethyl auristatin F, monomethyl auristatin E, paciltaxol, SN-38, calicheamicin, anthramycin, abbeymycin, chicamycin, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin prothracarcin, sibanomicin, sibiromycin, tomamycin, mertansine, emtansine, irinotecan, camptothecin, topotecan, silatecan, cositecan, Exatecan, Lurtotecan, gimatecan, Belotecan, and Rub
• each occurrence of M 3 is auristatin F, monomethyl auristatin F, monomethyl auristatin E, paciltaxol, SN-38, calicheamicin, anthramycin, abbeymycin, chicamycin, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin prothracarcin, sibanomicin, sibiromycin, tomamycin, mertansine, emtansine, irinotecan, camptothecin, topotecan, silatecan, cositecan, Exatecan, Lurtotecan, gimatecan, Belotecan, and Rubitecan.
• At least one occurrence of M 3 has the following structure: In some specific embodiments, each occurrence of M 3 has the following structure: . In certain embodiments, at least one occurrence of –L 10 - M 3 has the following structure: . In certain embodiments, each occurrence of –L 10 - M 3 has the following structure: . In certain embodiments, at least one occurrence of M 3 has the following structure: . In some specific embodiments, each occurrence of M 3 has the following structure: . In certain embodiments, at least one occurrence of –L 10 - M 3 has the following structure: . In certain embodiments, each occurrence of –L 10 - M 3 has the following structure: .
• 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-multicyclic 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, his-fluorophenyl-BODIPY, acridine, terrylene, sexiphenyl, porphyrin, benzopyrene, (fluorophenyl-dimethyl-difluorobora- diaza-indacene)phenyl, (bis-fluorophenyl-difluorobora-diaza-indacene)phenyl, quaterphenyl, bi-benzothiazole, ter-benzothiazole, bi-naphthyl, bi-anthracyl, squaraine, squarylium
• 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-1,8-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:
• Such an attachment may be made by reducing a disulfide bond of a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) with an appropriate reagent (e.g., TCEP) and coupling the resultant molecule to an appropriate linker reagent (e.g., 1,1'-(ethane-1,2-diyl)bis(1H-pyrrole-2,5-dione) which is commonly known as bis- maleimidoethane or "BMOE").
• an appropriate reagent e.g., TCEP
• an appropriate linker reagent e.g., 1,1'-(ethane-1,2-diyl)bis(1H-pyrrole-2,5-dione
• R 2 comprises the following structure: wherein: L a is a direct bond or C1-C6 alkylene.
• L a is a direct bond
• R 2 further comprises a covalent bond to an antibody (e.g., monoclonal antibody such as brentuximab, gemtuzumab, trastuzumab, inotuzumab, polatuzumab, enfortumab, trastuzumab, sacituzumab, belantamab, moxetumomab, etc.) or fragment thereof.
• an antibody e.g., monoclonal antibody such as brentuximab, gemtuzumab, trastuzumab, inotuzumab, polatuzumab, enfortumab, trastuzumab, sacituzumab, belantamab, moxetumomab, etc.
• R 2 comprises the following structure: wherein: A is an antibody (e.g., monoclonal antibody such as brentuximab, gemtuzumab, trastuzumab, inotuzumab, polatuzumab, enfortumab, trastuzumab, sacituzumab, belantamab, or moxetumomab).
• L a is a direct bond.
• R 2 has the following structure: wherein: x 13 is 0 or an integer greater than 0 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12).
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has m from 0 to 10. In certain embodiments, a compound of Structure (I), (Ia), (Ib), (Ic), (Id), or (Ie) has m being 0, 1, 2, 3, 4, or 5. In some embodiments, a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has n being 1, 2, 3, or 4.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has n being 1 or 2.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has p being 1, 2, 3, or 4.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has p being 1 or 2.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has m being 0, n being 1, and p being 2.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has m being 1, n being 1, and p being 2.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) has m being 5, n being 1, and p being 2.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) wherein at least each occurrence of R 6 and R 7 are F, R 8 is O, R 9 is H, and R 10 is H.
• a method of treating a disease or disorder comprising administering a therapeutically effective amount of , a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId), or the pharmaceutical composition of the compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId), to a subject in need thereof is disclosed.
• the disease or disorder is cancer which includes breast cancer, stomach cancer, lung cancer, ovarian cancer, lymphoma, and bladder cancer.
• a compound of Structure (III) has Auristatin F moiety (labeled as “AF”) and gemcitabine moiety switched compared to Structure (I), (Ia), (Ib), (Ic), or (Id).
• AF Auristatin F moiety
• gemcitabine moiety switched compared to Structure (I), (Ia), (Ib), (Ic), or (Id).
• the compound of Structure (III) with AF moiety and gemcitabine moiety switched is shown as compound I-13 on Table 2 and can be prepared according to the procedures described in the present disclosure.
• a compound of Structure (I), (Ia), (Ib), (Ic), (Id), (Ie), (III), (IIIa), (IIIb), (IIIc), or (IIId) is a compound selected from Table 2.
• Tables 2 and 3 were prepared according to the procedures set forth in the Examples. Pharmaceutical Compositions
• compositions comprising the compound according any one of the embodiments disclosed herein (e.g., a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId)) and a pharmaceutically acceptable carrier.
• the pharmaceutical composition comprises any one (or more) of the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) and a pharmaceutically acceptable carrier.
• the pharmaceutical composition is formulated for oral administration.
• the pharmaceutical composition is formulated for injection.
• the pharmaceutical compositions comprise a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) and an additional therapeutic agent (e.g., anticancer agent).
• an additional therapeutic agent e.g., anticancer agent
• Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration.
• parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injections.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is administered in a local rather than systemic manner, for example, via injection of the compound directly into an organ, often in a depot preparation or sustained release formulation.
• long acting formulations are administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection.
• the drug is delivered in a targeted drug delivery system, for example, in a liposome coated with organ-specific antibody. In such embodiments, the liposomes are targeted to and taken up selectively by the organ.
• the compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is provided in the form of a rapid release formulation, in the form of an extended release formulation, or in the form of an intermediate release formulation.
• the compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is administered topically.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) are effective over a wide dosage range.
• dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg per day are examples of dosages that are used in some embodiments.
• An exemplary dosage is 10 to 30 mg per day. The exact dosage will depend upon the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, and the preference and experience of the attending physician.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is administered in a single dose.
• administration will be by injection, e.g., intravenous injection, in order to introduce the agent quickly.
• other routes are used as appropriate.
• a single dose of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) may also be used for treatment of an acute condition.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is administered in multiple doses.
• dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, dosing is about once a month, once every two weeks, once a week, or once every other day.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) and another agent are administered together about once per day to about 6 times per day.
• administration of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) and an agent continues for less than about 7 days.
• the administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day.
• a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) is administered chronically on an ongoing basis, e.g., for the treatment of chronic effects.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) are administered in dosages. It is known in the art that due to inter-subject variability in compound pharmacokinetics, individualization of dosing regimen is necessary for optimal therapy. Dosing for a compound of the disclosure may be found by routine experimentation in light of the instant disclosure.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) are formulated into pharmaceutical compositions.
• pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
• compositions comprising a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s).
• the compounds described are administered as pharmaceutical compositions in which compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) are mixed with other active ingredients, as in combination therapy.
• the pharmaceutical compositions include one or more compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd).
• a pharmaceutical composition refers to a mixture of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients.
• the pharmaceutical composition facilitates administration of the compound to an organism.
• therapeutically effective amounts of compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) provided herein are administered in a pharmaceutical composition to a mammal having a disease, disorder or medical condition to be treated.
• the mammal is a human.
• therapeutically effective amounts vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used and other factors.
• one or more compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) is formulated in an aqueous solution.
• the aqueous solution is selected from, by way of example only, a physiologically compatible buffer, such as Hank’s solution, Ringer’s solution, or physiological saline buffer.
• one or more compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) is/are formulated for transmucosal administration.
• transmucosal formulations include penetrants that are appropriate to the barrier to be permeated.
• appropriate formulations include aqueous or non-aqueous solutions.
• such solutions include physiologically compatible buffers and/or excipients.
• compounds described herein are formulated for oral administration.
• Compounds described herein are formulated by combining the active compounds with, e.g., pharmaceutically acceptable carriers or excipients.
• the compounds described herein are formulated in oral dosage forms that include, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions and the like.
• pharmaceutical preparations for oral use are obtained by mixing one or more solid excipient with one or more of the compounds described herein, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
• Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as: for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate.
• disintegrating agents are optionally added. Disintegrating agents include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
• dosage forms such as dragee cores and tablets, are provided with one or more suitable coating.
• concentrated sugar solutions are used for coating the dosage form.
• the sugar solutions optionally contain additional components, such as by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
• Dyestuffs and/or pigments are also optionally added to the coatings for identification purposes. Additionally, the dyestuffs and/or pigments are optionally utilized to characterize different combinations of active compound doses.
• Oral dosage forms include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
• push-fit capsules contain the active ingredients in admixture with one or more filler.
• Fillers include, by way of example only, lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
• soft capsules contain one or more active compound that is dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oil, liquid paraffin, or liquid polyethylene glycol.
• stabilizers are optionally added.
• therapeutically effective amounts of at least one of the compounds described herein are formulated for buccal or sublingual administration.
• Formulations suitable for buccal or sublingual administration include, by way of example only, tablets, lozenges, or gels.
• the compounds described herein are formulated for parental injection, including formulations suitable for bolus injection or continuous infusion.
• formulations suitable for injection are presented in unit dosage form (e.g., in ampoules) or in multi-dose containers. Preservatives are, optionally, added to the injection formulations.
• the pharmaceutical compositions are formulated in a form suitable for parenteral injection as sterile suspensions, solutions or emulsions in oily or aqueous vehicles.
• Parenteral injection formulations optionally contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
• pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form.
• suspensions of the active compounds e.g., compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) are prepared as appropriate oily injection suspensions.
• Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.
• aqueous injection suspensions contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran.
• the suspension contains suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
• the active ingredient is in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
• compositions such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments.
• Such pharmaceutical compositions optionally contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.
• transdermal formulations employ transdermal delivery devices and transdermal delivery patches and can be lipophilic emulsions or buffered, aqueous solutions, dissolved and/or dispersed in a polymer or an adhesive.
• transdermal patches are constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.
• transdermal delivery of the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) is accomplished by means of iontophoretic patches and the like.
• transdermal patches provide controlled delivery of the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd).
• the rate of absorption is slowed by using rate-controlling membranes or by trapping the compound within a polymer matrix or gel.
• absorption enhancers are used to increase absorption.
• transdermal devices include absorbable pharmaceutically acceptable solvents that assist passage through the skin.
• transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound to the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) are formulated for administration by inhalation.
• Various forms suitable for administration by inhalation include, but are not limited to, aerosols, mists or powders.
• compositions of any of compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas).
• a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
• the dosage unit of a pressurized aerosol is determined by providing a valve to deliver a metered amount.
• capsules and cartridges of, such as, by way of example only, gelatin for use in an inhaler or insufflator is formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (IIId) are formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinylpyrrolidone, PEG, and the like.
• a low-melting wax such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with melted cocoa butter.
• compositions are formulated in any conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any pharmaceutically acceptable techniques, carriers, and excipients are optionally used as suitable.
• compositions comprising a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (inc), or (IIId) are manufactured in a conventional manner, such as, by way of example only, by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compression processes.
• compositions include at least one pharmaceutically acceptable carrier, diluent or excipient and at least one compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd), described herein as an active ingredient.
• the active ingredient is in free-acid or free-base form, or in a pharmaceutically acceptable salt form.
• the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), as well as active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein.
• compositions optionally include other medicinal or pharmaceutical agents, carriers, adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure, buffers, and/or other therapeutically valuable substances.
• Methods for the preparation of compositions comprising the compounds described herein include formulating the compounds with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-solid or liquid.
• Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories.
• Liquid compositions include solutions in which a compound is dissolved, emulsions comprising a compound, or a solution containing liposomes, micelles, or nanoparticles comprising a compound as disclosed herein.
• Semi-solid compositions include, but are not limited to, gels, suspensions and creams.
• the form of the pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for solution or suspension in a liquid prior to use, or as emulsions. These compositions also optionally contain minor amounts of nontoxic, auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and so forth.
• composition comprising at least one compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) illustratively takes the form of a liquid where the agents are present in solution, in suspension or both.
• a liquid composition includes a gel formulation.
• the liquid composition is aqueous.
• useful aqueous suspensions contain one or more polymers as suspending agents.
• Useful polymers include water-soluble polymers such as cellulosic polymers, e.g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers.
• Certain pharmaceutical compositions described herein comprise a mucoadhesive polymer, selected for example from carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginate and dextran.
• Useful pharmaceutical compositions also, optionally, include solubilizing agents to aid in the solubility of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd).
• solubilizing agent generally includes agents that result in formation of a micellar solution or a true solution of the agent.
• Certain acceptable nonionic surfactants for example polysorbate 80, are useful as solubilizing agents, as can ophthalmically acceptable glycols, polyglycols, e.g., polyethylene glycol 400, and glycol ethers.
• useful pharmaceutical compositions optionally include one or more pH adjusting agents or buffering agents, including acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; and buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride.
• acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids
• bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane
• buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride.
• acids, bases and buffers are included in an amount required to maintain pH of the composition in an acceptable range.
• compositions also, optionally, include one or more salts in an amount required to bring osmolality of the composition into an acceptable range.
• salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.
• compositions optionally include one or more preservatives to inhibit microbial activity.
• Suitable preservatives include mercury- containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.
• Still other useful compositions include one or more surfactants to enhance physical stability or for other purposes.
• Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.
• Still other useful compositions include one or more antioxidants to enhance chemical stability where required. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.
• aqueous suspension compositions are packaged in single-dose non-reclosable containers.
• multiple-dose reclosable containers are used, in which case it is typical to include a preservative in the composition.
• hydrophobic pharmaceutical compounds are employed. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also employed. In additional embodiments, the compounds described herein are delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization are employed.
• the formulations described herein comprise one or more antioxidants, metal chelating agents, thiol containing compounds and/or other general stabilizing agents.
• stabilizing agents include, but are not limited to: (a) about 0.5% to about 2% w/v glycerol, (b) about 0.1% to about 1% w/v methionine, (c) about 0.1% to about 2% w/v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v polysorbate 80, (g) 0.001% to about 0.05% w/v.
• polysorbate 20 (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (1) pentosan poly sulfate and other heparinoids, (m) divalent cations such as magnesium and zinc; or (n) combinations thereof.
• the concentration of one or more compounds provided in the pharmaceutical compositions is less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v.
• the concentration of one or more compounds is greater than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%, 4%,
• the concentration of one or more compounds is in the range from approximately 0.0001% to approximately 50%, approximately 0.001% to approximately 40 %, approximately 0.01% to approximately 30%, approximately 0.02% to approximately 29%, approximately 0.03% to approximately 28%, approximately 0.04% to approximately 27%, approximately 0.05% to approximately 26%, approximately 0.06% to approximately 25%, approximately 0.07% to approximately 24%, approximately 0.08% to approximately 23%, approximately 0.09% to approximately 22%, approximately 0.1% to approximately 21%, approximately 0.2% to approximately 20%, approximately 0.3% to approximately 19%, approximately 0.4% to approximately 18%, approximately 0.5% to approximately 17%, approximately 0.6% to approximately 16%, approximately 0.7% to approximately 15%, approximately 0.8% to approximately 14%, approximately 0.9% to approximately 12%, approximately 1% to approximately 10% w/w, w/v or v/v.
• the concentration of one or more compounds is in the range from approximately 0.001% to approximately 10%, approximately 0.01% to approximately 5%, approximately 0.02% to approximately 4.5%, approximately 0.03% to approximately 4%, approximately 0.04% to approximately 3.5%, approximately 0.05% to approximately 3%, approximately 0.06% to approximately 2.5%, approximately 0.07% to approximately 2%, approximately 0.08% to approximately 1.5%, approximately 0.09% to approximately 1%, approximately 0.1% to approximately 0.9% w/w, w/v or v/v.
• the amount of one or more compounds is equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g
• the amount of one or more compounds is more than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g,
• the amount of one or more compounds ranges from 0.0001 to 10 g, 0.0005 to 9 g, 0.001 to 8 g, 0.005 to 7 g, 0.01 to 6 g, 0.05 to 5 g, 0.1 to 4 g, 0.5 to 4 g, or 1 to 3 g.
• Certain compounds of the present disclosure are useful for treating disease (i.e., compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd)).
• Those compounds disclosed herein offer a targeted approach to drug delivery strategies.
• one embodiment provides a method of treating a disease (or the symptoms thereof) comprising administering to a mammal (e.g., a human) in need thereof a therapeutically effective amount of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd).
• the disclosure provides a method of treating solid tumors, multiple myeloma, gliomas, clear cell renal cell carcinoma, prostate cancer, ovarian cancer, non-small cell lung cancer, GI malignancies, acute lymphoblastic leukemia, acute myelogenous leukemia, renal cell carcinoma, colorectal carcinoma, epithelial cancers, pancreatic and gastric cancers, renal cell carcinoma, non- Hodgkin’s lymphoma, metastatic renal cell carcinoma, malignant mesothelioma, pancreatic, ovarian, and/or lung adenocarcinoma, B-cell malignancies, breast cancer, melanoma, recurrent multiple myeloma, small cell lung cancer, CD22-positive B cell malignancies, Hodgkin’s lymphoma/anaplastic large cell lymphoma, or HER2-positive breast cancer.
• GI malignancies acute lymphoblastic leukemia, acute myelogenous leukemia, renal cell carcinoma, color
• the disease is cancer.
• the cancer is breast cancer, non-Hodgkin's lymphoma, acute myeloid leukemia, multiple myeloma, gastric cancer, renal cell carcinoma, solid tumors, ovarian cancer, prostate cancer, colorectal cancer, pancreatic cancer, small cell lung cancer, diffuse large B-cell lymphoma, a neoplasm, urothelial cancer, ALL, CLL, glioblastoma, Hodgkin's lymphoma, lymphoma, mesothelioma, non-small cell lung cancer, recurrent head and neck cancer, or a combination thereof.
• Certain embodiments also relate to a method of treating a hyperproliferative disorder in a mammal (e.g., a human) that comprises administering to said mammal a therapeutically effective amount of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd), or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.
• a mammal e.g., a human
• said method relates to the treatment of cancer such as acute myeloid leukemia, cancer in adolescents, adrenocortical carcinoma childhood, AIDS-related cancers (e.g., Lymphoma and Kaposi's Sarcoma), anal cancer, appendix cancer, astrocytomas, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumors, Burkitt lymphoma, carcinoid tumor, atypical teratoid, embryonal tumors, germ cell tumor, primary lymphoma, cervical cancer, childhood cancers, chordoma, cardiac tumors, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myleoproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal
• cancer such
• said method relates to the treatment of a non-cancerous hyperproliferative disorder such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).
• a non-cancerous hyperproliferative disorder such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).
• Certain particular embodiments provide methods for treatment of lung cancers, the methods comprise administering an effective amount of any of the above described compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) (or a pharmaceutical composition comprising the same) to a subject in need thereof.
• the lung cancer is a non-small cell lung carcinoma (NSCLC), for example adenocarcinoma, squamous-cell lung carcinoma or large-cell lung carcinoma.
• the lung cancer is a small cell lung carcinoma.
• Other lung cancers treatable with the disclosed compounds include, but are not limited to, glandular tumors, carcinoid tumors and undifferentiated carcinomas.
• A is an antibody or a cell surface receptor antagonist.
• EGFR epidermal growth factor receptor
• HGFR hepatocyte growth factor receptor
• IGFR insulin-like growth factor receptor
• folate a folate
• MET a MET inhibitor
• trastuzumab antibodies such as trastuzumab.
• the method further comprises inducing apoptosis.
• the method of treatment comprises treating a tumor having tumor cells with tumor cell receptors.
• the tumor cells have receptors ranging from 1,000 to 100,000, from 1,000 to 50,000, from 1,000 to 25,000 receptors, 1,000 to 10,000 receptors per cell.
• the tumor cells have about 1,000, about 10,000, or less than 100,000 receptors per cell.
• Further therapeutic agents that can be combined with a compound of the disclosure are found in Goodman and Gilman’s "The Pharmacological Basis of Therapeutics" Tenth Edition edited by Hardman, Limbird and Gilman or the Physician’s Desk Reference, both of which are incorporated herein by reference in their entirety.
• the compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd) described herein can be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated.
• the one or more compounds of the disclosure will be co- administered with other agents as described above.
• the compounds described herein are administered with the second agent simultaneously or separately.
• This administration in combination can include simultaneous administration of the two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, a compound described herein and any of the agents described above can be formulated together in the same dosage form and administered simultaneously.
• a compound of the disclosure and any of the agents described above can be simultaneously administered, wherein both the agents are present in separate formulations.
• a compound of the present disclosure can be administered just followed by and any of the agents described above, or vice versa.
• a compound of the disclosure and any of the agents described above are administered a few minutes apart, or a few hours apart, or a few days apart.
• the method further comprises administering an additional therapeutic agent selected from the group consisting of an antineoplastic agent, an enediyne antitumor antibiotic, a maytansinoid, a topoisomerase inhibitor, a kinase inhibitor, an anthracycline, and EGFR inhibitor, an alkylating agent and combinations thereof.
• an additional therapeutic agent selected from the group consisting of an antineoplastic agent, an enediyne antitumor antibiotic, a maytansinoid, a topoisomerase inhibitor, a kinase inhibitor, an anthracycline, and EGFR inhibitor, an alkylating agent and combinations thereof.
• the method further comprises administering an additional therapeutic agent selected from the group consisting of an antineoplastic agent, an enediyne antitumor antibiotic, a maytansinoid, a topoisomerase inhibitor, a kinase inhibitor, an anthracychne, and EGFR inhibitor, an alkylating agent and combinations thereof.
• an additional therapeutic agent selected from the group consisting of an antineoplastic agent, an enediyne antitumor antibiotic, a maytansinoid, a topoisomerase inhibitor, a kinase inhibitor, an anthracychne, and EGFR inhibitor, an alkylating agent and combinations thereof.
• the additional therapeutic agent comprises auristatin F, monomethyl auristatin F, monomethyl auristatin E, paciltaxol, SN-38, calicheamicin, anthramycin, abbeymycin, chicamycin, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin prothracarcin, sibanomicin, sibiromycin, tomamycin, mertansine, emtansine, irinotecan, camptothecin, topotecan, silatecan, cositecan, Exatecan, Lurtotecan, gimatecan, Belotecan, and Rubitecan.
• auristatin F monomethyl auristatin F
• monomethyl auristatin E monomethyl auristatin E
• paciltaxol SN-38
• calicheamicin anthramycin, abbeymycin, chica
• compositions comprising any of the foregoing compounds and one or more analyte molecules (e.g., biomolecules) are provided in various other embodiments.
• use of such compositions in analytical methods for detection of the one or more analyte molecules is also provided.
• R 2 is a linker comprising a covalent linkage to an analyte molecule, such as a biomolecule.
• a linker comprising a covalent linkage to an analyte molecule, such as a biomolecule.
• a biomolecule such as a biomolecule.
• the biomolecule is an enzyme, receptor, receptor ligand, antibody, glycoprotein, aptamer, or prion.
• R 2 is a linker comprising a covalent linkage to a solid support such as a microparticle (e.g., controlled pore glass or polystyrene beads).
• a microparticle e.g., controlled pore glass or polystyrene beads
• the microparticle is a polymeric bead or non-polymeric bead.
• embodiments of the compounds of structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), and (IIId) find utility in various disciplines and methods, including but not limited to: imaging in endoscopy procedures for identification of cancerous and other tissues; single-cell and/or single molecule analytical methods, for example detection of polynucleotides with little or no amplification; cancer imaging, for example by conjugating a compound of structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), and (IIId) to an antibody or sugar or other moiety that preferentially binds cancer cells; imaging in surgical procedures; binding of histones for identification of various diseases; drug delivery in dental work and other procedures.
• Suitable protecting groups include hydroxy, amino, mercapto and carboxylic acid.
• Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (for example, t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethyl silyl), tetrahydropyranyl, benzyl, and the like.
• Suitable protecting groups for amino, amidino and guanidino include /-butoxy carbonyl, benzyloxycarbonyl, and the like.
• Suitable protecting groups for mercapto include -C(O)-R" (where R" is alkyl, aryl or arylalkyl), /?-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.
• DNA synthesis methodology can be applied to build compounds of Structure (I) and (III).
• Monomers e.g., phosphoramidite monomers
• Monomers can be purchased commercially (e.g., from ChemGenes Corporation, Wilmington Mass.) or synthesized using methods described herein (see, e.g., Examples 1-3).
• 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., G 1 of General Reaction Scheme I).
• 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.
• Step 3 OXIDATION
• Step 4 CAPPING
• 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 chemotherapeutic agent
• Reaction Scheme I illustrates a method for preparation of phosphoramidite intermediates useful for preparation of compounds of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), or (Illd).
• G 1 represents a desired moiety containing a carboxylic acid functional group (e.g., a drug moiety such as Auristatin F, gemcitabine, capecitabine, or SN 38),
• L represents a bivalent linker moiety (e.g., an alkylene, or alkylene ether),
• X represents a leaving group (e.g., halo such as Cl), and
• PG represents a protecting group (e.g., 4,4'- dimethoxy triphenylmethyl).
• Step 1 of Reaction Scheme I starts with an activation of the carboxylic acid functional group of the first compound shown using known reagents under basic conditions (e.g., HATU and DIPEA in DMF).
• the activated acid is then reacted with an amine to provide the reaction product of Step 1.
• the resulting diol is then protected under standard conditions (e.g., 4,4'-dimethoxytriphenylmethyl chloride and pyridine).
• a diol such as gemcitabine starts from step 2.
• the protected product is then reacted with 3-((chloro(diisopropylamino)phosphaneyl)oxy)propanenitrile (or other appropriate reagent) to yield a desired compound of Structure (II) as shown above.
• the resultant compound of Structure (II) can then be used to synthesize a desired compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), and (Illd) by reaction under well-known (automated) DNA synthesis conditions.
• additional repeat units may be incorporated to achieve a final compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb), (IIIc), and (Illd).
• compounds having the following structure may be used: wherein: L is a desired linker moiety (e.g., including PEG or dye-containing moiety).
• the following compound may be used in the synthesis of a compound of Structure (I), (la), (lb), (Ic), (Id), (le), (III), (Illa), (Illb),
• 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-C 18 column held at 45 °C, employing an acetonitrile / water mobile phase gradient.
• Gencitabine (0.5057 mmole theoretical) is added to a dried round bottom flask, under inert gas blanket, with magnetic stir bar, followed by addition of pyridine, anhydrous (5.06 mL). The reaction flask is then transferred to an ice water bath (0°C) and allowed to cool with mixing until thermally equalized (approximately 10 minutes). Then, 4,4'-Dimethoxytrityl chloride (0.257 g, 0.759 mmole) is added to the cooled mixture with continuous mixing under inert gas. The reaction mixture is allowed to warm to room temperature then sampled for TLC analysis.
• the separated organic phase is washed with sodium chloride (5.06 mL, saturated aq.) and separated.
• the separated organic phase is dried over sodium sulfate, anhydrous and the sodium sulfate filtered off.
• the product containing organic phase is sampled for TLC and LC-UV/MS analysis. Solvent is removed by rotary evaporation, resulting in crude DMT protected gemcitabine.
• This crude material is then combined with crude material from a small- scale test reaction.
• the combined crude material is purified by silica gel flash chromatography, dichloromethane / methanol mobile phase, product containing fractions are pooled and solvent is removed by rotary evaporation, and then placed on vacuum line for at least 24 hours to yield DMT protected gemcitabine.
• the purified DMT protected gemcitabine (0.226 mmole), dried under vacuum for at least 24 hours is dissolved in di chloromethane (2.26 mL), under inert gas blanket, with magnetic stir bar, followed by addition of DIPEA (0.117 g), and then addition of Cl-Phos. (0.107 g).
• the reaction is allowed to mix for approximately 15 minutes and then sampled for TLC analysis (TLC showed reaction completion).
• TLC showed reaction completion.
• the reaction mixture is washed by adding directly to sodium bicarbonate (2.26 mL, saturated aq.) and organic phase separated, repeated one time. The organic phases are combined and dried over sodium sulfate, anhydrous, and then the sodium sulfate filtered off.
• the product containing organic phase is sampled for TLC and LC-UV/MS analysis. Then, di chloromethane is stripped off by rotary evaporation and proceeded to purification without crude weight. This crude material is then combined with crude material from a small-scale test reaction. The combined crude material is purified by silica gel solid phase extraction, di chloromethane/ methanol / tnethylamine mobile phase, product containing fractions are pooled. The mobile phase is stripped off by rotary evaporation, and then placed on vacuum line for at least 24 hours to yield DMT protected gemcitabine phosphoramidite. Alternatively, the DMT protected gemcitabine phosphoramidite can be purchased from Glen Research and used as is with no additional purification.
• the filtrate is removed and a third 10 mL 100 mM NaOH aliquot is added to the retentate.
• the setup is centrifuged as before and the filtrate removed.
• a fourth 10 mL 100 mM NaOH aliquot is added to the retentate and centrifuged as before.
• the filtrate is removed and 10 mL of water are added to the filtration setup.
• the mixture is centrifuged as before.
• the retentate is removed, the filtration vessel is washed with water and the rinseates are added to the final volume (3.5 mL).
• the desired product is confirmed by LC-MS and absorbance is used to determine concentration.
• Analytical LC-UV 495nm chromatogram showed 62% target product I-2 by total peak area or related peak area identified by MS (expected molecular weight 10755.1 and observed molecular weight 70760.7 was found).
• Analytical LC-UV 495nm chromatogram showed 65% target product I-6 by total peak area or related peak area identified by MS (expected molecular weight 5730.7 and observed molecular weight 5734.1 was found).
• Analytical LC-UV 266nm chromatogram showed 22% target product I-10 by total peak area or related peak area identified by MS (expected molecular weight 8722.8 and observed molecular weight 8730.1 was found).
• the mal eimide functionalized Compound I-1 is prepared according to the method described in Example 1.
• a trastuzumab antibody is treated with bis-maleimidoethane ("BMOE") to reduce disulfide bonds.
• BMOE bis-maleimidoethane
• the reduced antibody is reacted with Compound I-1 in a 5:1 molar ratio of polymer to antibody.
• the reaction results in a final product having a polymer to antibody ratio of 1 : 1 as detected by size exclusion chromatography.
• anti-CD33, anti-CD70, or anti- CD123 may be used with bis-maleimidoethane ("BMOE”) to reduce disulfide bonds.
• I- 2, I-6, and I-9 ADCs were prepared according to EXAMPLE 4 procedure and shown below:
• Other compounds disclosed herein (I-1, I-3 - I-5, I-7 - I-8, and I- 10 - I- 22) can be conjugated with antibody by the same method described above to generate other ADCs.
• antibody trastuzumab is used in EXAMPLE 4 demonstrating conjugation between compounds disclosed herein and an antibody, it is used for merely an illustration and can include other antibodies such as brentuximab, gemtuzumab, trastuzumab, inotuzumab, polatuzumab, enfortumab, trastuzumab, sacituzumab, belantamab, or moxetumomab.
• Compounds I-2, I-6, and I-9 were prepared on the DNA synthesizer as disclosed in the present disclosure. Compounds I-2, I-6, and I-9 were activated and conjugated to the commercial antibody Trastuzumab. As shown in FIGs. I-2, compound I-2 ADC (labeled as “ADC: 073-128-R3a”) was more potent and cytotoxic than other ADCs (I-9 ADC labeled as “ADC: 073-128-R1” and I-6 ADC labeled as ADC: 073-128-R2”) and its component parts including Auristatin F alone (labeled as “AF”) and Trastuzumab alone (labeled as “Herceptin”).
• ADC ADC
• AF Auristatin F alone
• Trastuzumab alone labeleled as “Herceptin”.
• Compound I-2 ADC was found to be potent and selective versus cell lines expressing the Her2 antigen.
• Compound I-2 ADC (labeled as “ADC: 073-128-R3a”) and compound 1-2 ADC (labeled as “ADC: 073-128-R3b”) differ by average number of polymers attached per antibody (degree of labeling, DOL).
• ADC: 073-128-R3a has a DOL of 1.7
• ADC: 073-128-R3b has a DOL of 0.9.

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