[go: up one dir, main page]

WO2024167952A1 - Médicaments à base d'organophosphate clivables par oxydation - Google Patents

Médicaments à base d'organophosphate clivables par oxydation Download PDF

Info

Publication number
WO2024167952A1
WO2024167952A1 PCT/US2024/014654 US2024014654W WO2024167952A1 WO 2024167952 A1 WO2024167952 A1 WO 2024167952A1 US 2024014654 W US2024014654 W US 2024014654W WO 2024167952 A1 WO2024167952 A1 WO 2024167952A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
alkyl
hydrogen
group
substituted
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2024/014654
Other languages
English (en)
Inventor
Paul E. Floreancig
Brittany KLOOTWYK
Amy E. RYAN
Alexander Deiters
Arbil LOPEZ
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Pittsburgh
Original Assignee
University of Pittsburgh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Pittsburgh filed Critical University of Pittsburgh
Publication of WO2024167952A1 publication Critical patent/WO2024167952A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C07F9/65586—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system at least one of the hetero rings does not contain nitrogen as ring hetero atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/657163—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms the ring phosphorus atom being bound to at least one carbon atom
    • C07F9/657181—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms the ring phosphorus atom being bound to at least one carbon atom the ring phosphorus atom and, at least, one ring oxygen atom being part of a (thio)phosphonic acid derivative
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/6574—Esters of oxyacids of phosphorus
    • C07F9/65744—Esters of oxyacids of phosphorus condensed with carbocyclic or heterocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H23/00—Compounds containing boron, silicon or a metal, e.g. chelates or vitamin B12

Definitions

  • H 2 O 2 Hydrogen peroxide's unique reactivity properties and importance in these conditions have resulted in its utilization to initiate a number of processes in biological and materials chemistry.
  • H 2 O 2 is an attractive agent for initiating prodrug unraveling in many cases because it is small and can access sterically hindered sites in structures that are inaccessible to enzymes, which are commonly utilized for this purpose.
  • a compound, or a pharmaceutically acceptable salt thereof having a structure of: Formula I wherein R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; R 3 is O or S; R 4 is hydrogen, alkyl, aryl, or silyl; R 5 is hydrogen, alkyl, aryl, or silyl; and Z is alkyl, aryl, or a cargo-containing group; or a compound, or a pharmaceutically acceptable salt thereof, having a structure of: OR1 R3 wherein R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; R 3 is O or S; R 4 is hydrogen, alkyl, aryl or silyl; R 5 is hydrogen, alkyl, ary
  • a compound, or a pharmaceutically acceptable salt thereof having a structure of: wherein X is O or N; when X is O, R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl, or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; when X is N, R 1 and R 2 together form a cyclic structure; R 3 is O or S; R 4 is hydrogen, alkyl, aryl, or silyl; R 5 is hydrogen, alkyl, aryl, or silyl; and is a cyclic structure that includes a cargo-containing group.
  • FIGS.1A-1B Time course of oxidative phosphate release.
  • FIG.1A Cleavage to release a phosphodiester.
  • FIG.1B Cleavage to release a phosphomonoester via a phosphodiester intermediate. The reactions were run in triplicate and error bars represent standard deviations from the mean.
  • FIG.2A ATP consumption through the phosphorylation of AZT monophosphate to form AZT diphosphate.
  • FIG.2B Workflow of assessing H2O2-dependent phosphorylation of 37 by thymidine monophosphate kinase through quantification of ATP consumption.
  • FIGS.3A-3C Timecourse analysis of ATP stability.
  • FIG.3A Validation of TMPK-HA expression in HEK293T cells and pulldown from cell lysates.
  • B bead eluants. Timecourse quantification of ATP concentration over the course of 4 hours in the absence or presence of H 2 O 2 (60 ⁇ M), boronate-AZTMP (2 ⁇ M), or boronate AZTMP preincubated with H 2 O 2 .
  • FIGS.4A-4D Oxidative control of 36 and subsequent phosphorylation of 37 to 39, analyzed by a luciferase assay. Timecourse quantification of ATP concentration over the course of 2 hours in the absence or presence of immobilized TMPK-HA and 2 ⁇ M of FIG.4A) 37 (positive control), FIG.4B) 38 (negative control), FIG.4C) the boronate-AZTMP 36 activated with H 2 O 2 and D) the boronate-AZTMP 36 activated in the absence of H 2 O 2 .
  • “Lower acyloxy” groups contain one to six carbon atoms.
  • Administration as used herein is inclusive of administration by another person to the subject or self-administration by the subject.
  • Alkenyl refers to a cyclic, branched or straight chain group containing only carbon and hydrogen, and contains one or more double bonds that may or may not be conjugated. Alkenyl groups may be unsubstituted or substituted.
  • “Lower alkenyl” groups contain one to six carbon atoms.
  • alkyl refers to a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like.
  • a “lower alkyl” group is a saturated branched or unbranched hydrocarbon having from 1 to 6 carbon atoms. Preferred alkyl groups have 1 to 4 carbon atoms.
  • Alkyl groups may be “substituted alkyls” wherein one or more hydrogen atoms are substituted with a substituent such as halogen, cycloalkyl, alkoxy, amino, hydroxyl, aryl, alkenyl, or carboxyl.
  • a lower alkyl or (C 1 -C 6 )alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl;
  • (C 3 -C 6 )cycloalkyl can be cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;
  • (C 3 -C 6 )cycloalkyl(C 1 -C 6 )alkyl can be cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2-cyclopentylethyl, or 2-cyclohexylethyl;
  • (C 1 -C 6 )alkoxy can be methoxy, ethoxy, prop
  • An “analog” is a molecule that differs in chemical structure from a parent compound, for example a homolog (differing by an increment in the chemical structure or mass, such as a difference in the length of an alkyl chain or the inclusion of one of more isotopes), a molecular fragment, a structure that differs by one or more functional groups, or a change in ionization.
  • An analog is not necessarily synthesized from the parent compound.
  • a derivative is a molecule derived from the base structure.
  • An “animal” refers to living multi-cellular vertebrate organisms, a category that includes, for example, mammals and birds.
  • mammal includes both human and non-human mammals.
  • subject includes both human and non-human subjects, including birds and non-human mammals.
  • Illustrative non-human mammals include animal models (such as mice), non-human primates, companion animals (such as dogs and cats), livestock (such as pigs, sheep, cows), as well as non-domesticated animals, such as the big cats.
  • subject applies regardless of the stage in the organism’s life-cycle.
  • the term subject applies to an organism in utero or in ovo, depending on the organism (that is, whether the organism is a mammal or a bird, such as a domesticated or wild fowl).
  • aralkyl refers to an alkyl group wherein an aryl group is substituted for a hydrogen of the alkyl group.
  • An example of an aralkyl group is a benzyl group.
  • Aryl refers to a monovalent unsaturated aromatic carbocyclic group having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl), which can optionally be unsubstituted or substituted.
  • a “heteroaryl group,” is defined as an aromatic group that has at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, and phosphorous.
  • Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and the like.
  • the aryl or heteroaryl group can be substituted with one or more groups including, but not limited to, alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ester, ketone, aldehyde, hydroxy, carboxylic acid, or alkoxy, or the aryl or heteroaryl group can be unsubstituted.
  • Inhibiting refers to inhibiting the full development of a disease or condition. Inhibiting also refers to any quantitative or qualitative reduction in biological activity or binding, relative to a control.
  • Phosphate refers to the group –O-P(O)(OR’) 2 , where each -OR’ independently is –OH; - O-aliphatic, such as –O-alkyl or –O-cycloalkyl; -O-aromatic, including both -O-aryl and -O- heteroaryl; –O-aralkyl; or -OR’ is –O-M + , where M + is a counter ion with a single positive charge.
  • Each M + may be an alkali ion, such as K + , Na + , Li + ; an ammonium ion, such as + N(R”) 4 where each R” independently is H, aliphatic, heterocyclyl or aryl; or an alkaline earth ion, such as [Ca 2+ ]0.5, [Mg 2+ ] 0.5 , or [Ba 2+ ] 0.5 .
  • Phosphonooxyalkyl refers to the group –alkyl-phosphate, such as, for example, -CH 2 OP(O)(OH) 2 , or a salt thereof, such as -CH 2 OP(O)(O-Na + ) 2 , and (((dialkoxyphosphoryl)oxy)alkyl) refers to the dialkyl ester of a phosphonooxyalkyl group, such as, for example, -CH 2 OP(O)(O-tert-butyl) 2 .
  • subject includes both human and non-human subjects, including birds and non-human mammals, such as non-human primates, companion animals (such as dogs and cats), livestock (such as pigs, sheep, cows), as well as non-domesticated animals, such as the big cats.
  • non-human mammals such as non-human primates, companion animals (such as dogs and cats), livestock (such as pigs, sheep, cows), as well as non-domesticated animals, such as the big cats.
  • livestock such as pigs, sheep, cows
  • non-domesticated animals such as the big cats.
  • subject applies regardless of the stage in the organism’s life-cycle. Thus, the term subject applies to an organism in utero or in ovo, depending on the organism (that is, whether the organism is a mammal or a bird, such as a domesticated or wild fowl).
  • “Substituted” or “substitution” refers to replacement of a hydrogen atom of a molecule or an R-group with one or more
  • the term “optionally- substituted” or “optional substituent” as used herein refers to a group which may or may not be further substituted with 1, 2, 3, 4 or more groups, preferably 1, 2 or 3, more preferably 1 or 2 groups.
  • the substituents may be selected, for example, from C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3- 8 cycloalkyl, hydroxyl, oxo, C 1-6 alkoxy, aryloxy, C 1-6 alkoxyaryl, halo, C 1-6 alkylhalo (such as CF 3 and CHF 2 ), C 1-6 alkoxyhalo (such as OCF 3 and OCHF 2 ), carboxyl, esters, cyano, nitro, amino, substituted amino, disubstituted amino, acyl, ketones, amides, aminoacyl, substituted amides, disubstituted amides, thiol, alkylthio,
  • N-heterocycles may also include but are not limited to C1-6alkyl i.e. N-C 1-3 alkyl, more preferably methyl, particularly N-methyl.
  • a "therapeutically effective amount” refers to a quantity of a specified agent sufficient to achieve a desired effect in a subject being treated with that agent. Ideally, a therapeutically effective amount of an agent is an amount sufficient to inhibit or treat the disease or condition without causing a substantial cytotoxic effect in the subject. The therapeutically effective amount of an agent will be dependent on the subject being treated, the severity of the affliction, and the manner of administration of the therapeutic composition.
  • Treatment refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop.
  • the term “ameliorating,” with reference to a disease or pathological condition refers to any observable beneficial effect of the treatment.
  • the beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art that are specific to the particular disease.
  • treating a disease refers to inhibiting the full development of a disease, for example, in a subject who is at risk for a disease.
  • a “prophylactic” treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing a pathology or condition, or diminishing the severity of a pathology or condition.
  • “Pharmaceutical compositions” are compositions that include an amount (for example, a unit dosage) of one or more of the disclosed compounds together with one or more non-toxic pharmaceutically acceptable additives, including carriers, diluents, and/or adjuvants, and optionally other biologically active ingredients.
  • compositions can be prepared by standard pharmaceutical formulation techniques such as those disclosed in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA (19th Edition).
  • pharmaceutically acceptable salt or ester refers to salts or esters prepared by conventional means that include salts, e.g., of inorganic and organic acids, including but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, malic acid, acetic acid, oxalic acid, tartaric acid, citric acid, lactic acid, fumaric acid, succinic acid, maleic acid, salicylic acid, benzoic acid, phenylacetic acid, mandelic acid and the like.
  • “Pharmaceutically acceptable salts” of the presently disclosed compounds also include those formed from cations such as sodium, potassium, aluminum, calcium, lithium, magnesium, zinc, and from bases such as ammonia, ethylenediamine, N-methyl-glutamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N- benzylphenethylamine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane, and tetramethylammonium hydroxide.
  • bases such as ammonia, ethylenediamine, N-methyl-glutamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N- benzylphenethylamine, diethylamine, piperazine
  • any chemical compound recited in this specification may alternatively be administered as a pharmaceutically acceptable salt thereof.
  • “Pharmaceutically acceptable salts” are also inclusive of the free acid, base, and zwitterionic forms. Descriptions of suitable pharmaceutically acceptable salts can be found in Handbook of Pharmaceutical Salts, Properties, Selection and Use, Wiley VCH (2002). When compounds disclosed herein include an acidic function such as a carboxy group, then suitable pharmaceutically acceptable cation pairs for the carboxy group are well known to those skilled in the art and include alkaline, alkaline earth, ammonium, quaternary ammonium cations and the like. Such salts are known to those of skill in the art. For additional examples of “pharmacologically acceptable salts,” see Berge et al., J.
  • “Pharmaceutically acceptable esters” includes those derived from compounds described herein that are modified to include a carboxyl group.
  • An in vivo hydrolysable ester is an ester, which is hydrolysed in the human or animal body to produce the parent acid or alcohol.
  • esters thus include carboxylic acid esters in which the non-carbonyl moiety of the carboxylic acid portion of the ester grouping is selected from straight or branched chain alkyl (for example, methyl, n-propyl, t-butyl, or n-butyl), cycloalkyl, alkoxyalkyl (for example, methoxymethyl), aralkyl (for example benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for example, phenyl, optionally substituted by, for example, halogen, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy) or amino); sulphonate esters, such as alkyl- or aralkylsulphonyl (for example, methanesulphonyl); or amino acid esters (for example, L-valyl or L-isoleucyl).
  • alkyl for example, methyl, n-propyl, t
  • a “pharmaceutically acceptable ester” also includes inorganic esters such as mono-, di-, or tri- phosphate esters.
  • any alkyl moiety present advantageously contains from 1 to 18 carbon atoms, particularly from 1 to 6 carbon atoms, more particularly from 1 to 4 carbon atoms.
  • Any cycloalkyl moiety present in such esters advantageously contains from 3 to 6 carbon atoms.
  • Any aryl moiety present in such esters advantageously comprises a phenyl group, optionally substituted as shown in the definition of carbocycylyl above.
  • esters thus include C 1 -C 22 fatty acid esters, such as acetyl, t-butyl or long chain straight or branched unsaturated or omega-6 monounsaturated fatty acids such as palmoyl, stearoyl and the like.
  • Alternative aryl or heteroaryl esters include benzoyl, pyridylmethyloyl and the like any of which may be substituted, as defined in carbocyclyl above.
  • Additional pharmaceutically acceptable esters include aliphatic L-amino acid esters such as leucyl, isoleucyl and especially valyl.
  • salts of the compounds are those wherein the counter-ion is pharmaceutically acceptable.
  • salts of acids and bases which are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.
  • the pharmaceutically acceptable acid and base addition salts as mentioned hereinabove are meant to comprise the therapeutically active non-toxic acid and base addition salt forms which the compounds are able to form.
  • the pharmaceutically acceptable acid addition salts can conveniently be obtained by treating the base form with such appropriate acid.
  • Appropriate acids comprise, for example, inorganic acids such as hydrohalic acids, e.g.
  • hydrochloric or hydrobromic acid sulfuric, nitric, phosphoric and the like acids; or organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic, fumaric, malic (i.e.
  • salt forms can be converted by treatment with an appropriate base into the free base form.
  • the compounds containing an acidic proton may also be converted into their non-toxic metal or amine addition salt forms by treatment with appropriate organic and inorganic bases.
  • Appropriate base salt forms comprise, for example, the ammonium salts, the alkali and earth alkaline metal salts, e.g.
  • addition salt as used hereinabove also comprises the solvates which the compounds described herein are able to form. Such solvates are for example hydrates, alcoholates and the like.
  • quaternary amine as used hereinbefore defines the quaternary ammonium salts which the compounds are able to form by reaction between a basic nitrogen of a compound and an appropriate quaternizing agent, such as, for example, an optionally substituted alkylhalide, arylhalide or arylalkylhalide, e.g. methyliodide or benzyliodide.
  • an appropriate quaternizing agent such as, for example, an optionally substituted alkylhalide, arylhalide or arylalkylhalide, e.g. methyliodide or benzyliodide.
  • Other reactants with good leaving groups may also be used, such as alkyl trifluoromethanesulfonates, alkyl methanesulfonates, and alkyl p-toluenesulfonates.
  • a quaternary amine has a positively charged nitrogen.
  • Counterions include chloro, bromo, iodo, trifluoroacetate and acetate.
  • the counterion of choice can be introduced using ion exchange resins.
  • Particular examples of the presently disclosed compounds may include one or more asymmetric centers; thus the compounds described can exist in different stereoisomeric forms. Accordingly, compounds and compositions may be provided as individual pure enantiomers or as stereoisomeric mixtures, including racemic mixtures.
  • the compounds disclosed herein may be synthesized in or may be purified to be in substantially enantiopure form, such as in a 90% enantiomeric excess, a 95% enantiomeric excess, a 97% enantiomeric excess or even in greater than a 99% enantiomeric excess, such as in enantiopure form.
  • the compounds can be isolated as a single isomer or as mixture of isomers.
  • All tautomers of the compounds are also considered part of the disclosure.
  • the presently disclosed compounds also include all isotopes of atoms present in the compounds, which can include, but are not limited to, deuterium, tritium, 18 F, etc.
  • Compounds The phosphate group is an essential component of a wide range of biomolecules. The challenges of employing phosphorylated molecules as therapeutic agents are illustrated by nucleoside/nucleotide anti-cancer and anti-viral agents. Nucleoside analogs must be converted to triphosphates through sequential phosphorylation steps to disrupt polymerase-catalyzed nucleic acid chain elongation.
  • nucleobases and/or to the sugar unit can lead to inefficient phosphorylation, thereby diminishing therapeutic efficacy.
  • Nucleotide analogs subvert the initial phosphorylation problem since the phosphate group is already present at the 5’-site of the structure. However, the charged group substantially diminishes cell permeability, thereby mitigating biological activity. This problem has been addressed through the use of phosphate prodrugs (see below), as seen in the clinical agents sofosbuvir (3) and remdesivir (4). These neutral agents enter cells and release nucleotide analogs through an esterase-initiated cascade.
  • phosphate prodrugs employ the esterase labile pivaloylmethyl (POM, 5) and S-acyl-2-thioethyl (SATE, 6) groups, while the cyclic phosphotriesters cyclo-Sal group (7) and HepDirect (8) are activated by acid and oxidation by cytochrome P4503A, respectively.
  • POM esterase labile pivaloylmethyl
  • SATE S-acyl-2-thioethyl
  • HepDirect (8) are activated by acid and oxidation by cytochrome P4503A, respectively.
  • the releasable active moiety is a bioactive moiety such as a therapeutically active moiety.
  • the compounds release the active moiety upon reaction with hydrogen peroxide.
  • the compounds are prodrugs that release the active moiety in vivo upon reaction with hydrogen peroxide.
  • the by-products of the active moiety release are non-toxic.
  • the compounds disclosed herein avoid the release of toxic quinone methides or acroleins that are generated upon the breakdown of other peroxide-sensitive prodrugs. Rather, the compounds disclosed herein release alcohols, ketones or aldehydes under oxidative conditions.
  • boronate oxidation triggers fragmentation reactions that release alcohols, aldehydes, and ketones, through the generation of boron enolates from boryl allyloxy (BAO) groups.
  • BAO boryl allyloxy
  • One embodiment is a phosphate group-release strategy based on the peroxide-mediated oxidation of borylated allylic phosphoesters.
  • Alkenylboronates can be incorporated into phosphotriesters to release phosphodiesters and phosphomonoesters upon exposure to hydrogen peroxide. The reactions are rapid and efficient, and proceed cleanly even when substrate and peroxide concentrations are low.
  • the protocol was applied to nucleotide release and demonstrated through the release of the 5’-monophosphate of AZT.
  • the nucleotide that was released upon reaction with hydrogen peroxide proved to be a suitable substrate for enzymatic phosphorylation, validating the compatibility of the process with biomolecular transformations.
  • the presence of elevated levels of hydrogen peroxide in numerous disease states indicates that this protocol will be applicable to the development of site-selective phosphate-containing prodrugs that offer advantages over systems that release phosphates through interactions with ubiquitous stimuli.
  • the compounds are substantially non-toxic to a living cell, and thus are suitable for reacting with H 2 O 2 in a living cell (in vivo, ex vivo, or in vitro), in the extracellular medium in which a living cell is cultured in vitro or ex vivo, or extracellularly in a multicellular organism.
  • the compound selectively reacts with hydrogen peroxide, compared to other ROS.
  • a subject compound reacts with hydrogen peroxide, and does not substantially react with ROS other than hydrogen peroxide, e.g., the compound does not substantially react with any of superoxide anion, nitric oxide, peroxyl radical, alkoxyl radical, hydroxyl radical, hypochlorous acid, and singlet oxygen.
  • the compound reacts (and cleaves) with endogenous hydrogen peroxide.
  • the compound reacts (and cleaves) with exogenous hydrogen peroxide (e.g., hydrogen peroxide generated by a hydrogen peroxide inducer administered to a subject).
  • the compound reacts (and cleaves) with endogenous hydrogen peroxide and exogenous hydrogen peroxide.
  • the hydrogen peroxide can be induced by administering radiation to the subject (e.g., cancer radiotherapy).
  • the compounds are self-immolative, e.g., compounds that respond to an external stimulus (e.g., a reactive oxygen species) to undergo a fragmentation or cleavage to release an active moiety.
  • R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group;
  • R 3 is O or S;
  • R 4 is hydrogen, alkyl, aryl, or silyl;
  • R 5 is hydrogen, alkyl, aryl, or silyl;
  • Z is alkyl, aryl, or a cargo-containing group.
  • R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group;
  • R 3 is O or S;
  • R 4 is hydrogen, alkyl, aryl or silyl;
  • R 5 is hydrogen, alkyl, aryl or silyl; and each Z is independently alkyl, aryl, or a cargo-containing group.
  • R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; R 3 is O or S; and each Z is independently alkyl, aryl, or a cargo-containing group.
  • R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; R 3 is O or S; and Z is alkyl, aryl, or a cargo-containing group.
  • R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group;
  • R 6 is a nucleobase;
  • X is hydrogen, hydroxyl, F, F 2 , N 3 , alkenyl, substituted alkenyl, or acyloxy;
  • Y is hydrogen, hydroxyl, F, F 2 , N 3 , alkenyl, substituted alkenyl, or acyloxy.
  • a compound, or a pharmaceutically acceptable salt thereof having a structure of: O 3 A R P Formula VI wherein X is O or N; when X is O, R 1 and R 2 are each independently hydrogen, alkyl, or substituted alkyl, or R 1 and R 2 together form a boronic ester ring or a substituted boronic ester group; when X is N, R 1 and R 2 together form a cyclic structure; R 3 is O or S; R 4 is hydrogen, alkyl, aryl, or silyl; R 5 is hydrogen, alkyl, aryl, or silyl; and is a cyclic structure that includes a cargo-containing group.
  • R 1 and R 2 can each be independently selected from hydrogen, alkyl, or substituted alkyl; or R 1 and R 2 together can form a boronic ester ring or substituted boronic ester ring.
  • both R 1 and R 2 are hydrogen.
  • both R 1 and R 2 are alkyl, such as, for example, methyl, ethyl, propyl, isopropyl, and butyl.
  • R 1 and R 2 together form a boronic ester ring or substituted boronic ester ring.
  • R 1 and R 2 together form a boronic ester ring.
  • R 1 and R 2 together form a substituted boronic ester ring.
  • the -B(OR 1 )(OR 2 ) group is selected from: .
  • the -B(OR 1 )(OR 2 ) group is: .
  • R 3 is O.
  • R 4 and R 5 are each hydrogen.
  • R 4 is methyl and R 5 is hydrogen.
  • R 4 is hydrogen and R 5 is methyl.
  • Z is a cargo-containing group.
  • Illustrative cargo-containing groups for Z, and in formula VI for are groups that are included in phosphate-containing bioactive natural products and drugs.
  • Z is an amino acid or amino acid analog.
  • Z and in formula VI, includes a nucleoside.
  • the nucleoside may be bound to the phosphate group of the above formula I-IV via the sugar moiety of the nucleoside.
  • Illustrative nucleosides and nucleotide analogs include:
  • nucleosides and nucleotides include RX-3117, T-araC, 8-amino-adenosine, 8- chloro-adenosine, ARC, valopicitabine, elvucitabine, apricitabine, festinavir, 4’-thio-2’- deoxycytidine, 4’-thio-5-aza-2’-deoxycytidine, zebularine, UA911, pradefovir, cytarabine, INX- 08189, IDX-184, IDX-19368, CP-4126, CP-4200, CMX157, D07001-F4, YNK01, CMX001, didanosine, galidesivir, remdesivir, emtricitabine, zalcitabine, abacavir, entecavir, stavudine, zidovudine.
  • R 6 is a nucleobase derived from uridine, thymidine, cytidine, adenosine, guanosine, 2’-deoxycytidine, 2’-deoxyadenosine, 2’-deoxyguanosine or an analog thereof.
  • R 6 is adenine, guanine, cytosine, thymine, uracil, or an analog thereof.
  • an additional allyl-B(OR 1 )(OR 2 ) group as described above can be attached to the Z group.
  • the compounds are prodrugs.
  • a prodrug is an active or inactive compound that is modified chemically through in vivo physiological action (reaction with H 2 O 2 ) into an active compound following administration of the prodrug to a subject.
  • Locations in the body and conditions with elevated levels of H 2 O 2 are described herein and include toxic-metabolic encephalopathy (TME) and inflamed cells.
  • TME toxic-metabolic encephalopathy
  • Prodrugs preferably have excellent aqueous solubility, increased bioavailability and are readily metabolized into the active inhibitors in vivo.
  • the compound of formula V can cleave upon reaction with H 2 O 2 as shown below: Methods of Use and Compositions
  • the compounds disclosed herein are useful anti-cancer prodrugs or anti-viral prodrugs.
  • Neoplasms treatable by the presently disclosed compounds include all solid tumors, i.e., carcinomas and sarcomas.
  • Carcinomas include those malignant neoplasms derived from epithelial cells which tend to infiltrate (invade) the surrounding tissues and give rise to metastases.
  • Adenocarcinomas are carcinomas derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • Sarcoma broadly includes tumors whose cells are embedded in a fibrillar or homogeneous substance like embryonic connective tissue.
  • a solid tumor can be malignant, e.g. tending to metastasize and being life threatening, or benign.
  • solid tumors that can be treated include sarcomas and carcinomas such as, but not limited to: fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing’s tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma,
  • Illustrative cancers also include blood-borne cancers such as leukemia, myeloma, or lymphoma.
  • tumors comprising dysproliferative changes are treated or prevented in epithelial tissues such as those in the cervix, esophagus, and lung.
  • the presently disclosed methods provide for treatment of conditions known or suspected of preceding progression to neoplasia or cancer, in particular, where non-neoplastic cell growth consisting of hyperplasia, metaplasia, or most particularly, dysplasia has occurred.
  • Hyperplasia is a form of controlled cell proliferation involving an increase in cell number in a tissue or organ, without significant alteration in structure or function.
  • Metaplasia is a form of controlled cell growth in which one type of adult or fully differentiated cell substitutes for another type of adult cell. Metaplasia can occur in epithelial or connective tissue cells. Atypical metaplasia involves a somewhat disorderly metaplastic epithelium. Dysplasia is frequently a forerunner of cancer, and is found mainly in the epithelia; it is the most disorderly form of non-neoplastic cell growth, involving a loss in individual cell uniformity and in the architectural orientation of cells. Dysplastic cells often have abnormally large, deeply stained nuclei, and exhibit pleomorphism.
  • Dysplasia characteristically occurs where there exists chronic irritation or inflammation, and is often found in the cervix, respiratory passages, oral cavity, and gall bladder.
  • the presently disclosed methods are directed to a method for inhibiting cancer growth, including processes of cellular proliferation, invasiveness, and metastasis in biological systems.
  • the method is employed to inhibit or reduce cancer cell proliferation, invasiveness, metastasis, or tumor incidence in living animals, such as mammals.
  • cytotoxicity cell killing
  • the compounds disclosed herein can be used to induce cytotoxicity in cells of carcinomas of the prostate, breast, ovary, testis, lung, colon, or pancreas.
  • the compounds may be useful for treating hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), or herpes simplex virus (HSV).
  • HBV hepatitis B virus
  • HCV hepatitis C virus
  • HSV human immunodeficiency virus
  • HSV herpes simplex virus
  • the methods disclosed herein involve administering to a subject in need of treatment a pharmaceutical composition, for example a composition that includes a pharmaceutically acceptable carrier and a therapeutically effective amount of one or more of the compounds disclosed herein.
  • the compounds may be administered orally, parenterally (including subcutaneous injections (SC or depo-SC), intravenous (IV), intramuscular (IM or depo-IM), intrasternal injection or infusion techniques), sublingually, intranasally (inhalation), intrathecally, topically, ophthalmically, or rectally.
  • the pharmaceutical composition may be administered in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and/or vehicles.
  • the compounds are preferably formulated into suitable pharmaceutical preparations such as tablets, capsules, or elixirs for oral administration or in sterile solutions or suspensions for parenteral administration.
  • the compounds described above are formulated into pharmaceutical compositions using techniques and procedures well known in the art.
  • one or more of the disclosed compounds are mixed or combined with a suitable pharmaceutically acceptable carrier to prepare a pharmaceutical composition.
  • Pharmaceutical carriers or vehicles suitable for administration of the compounds provided herein include any such carriers known to be suitable for the particular mode of administration. Remington: The Science and Practice of Pharmacy, The University of the Sciences in Philadelphia, Editor, Lippincott, Williams, & Wilkins, Philadelphia, PA, 21 st Edition (2005), describes exemplary compositions and formulations suitable for pharmaceutical delivery of the compounds disclosed herein.
  • the compounds may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.
  • the resulting mixture may be a solution, suspension, emulsion, or the like.
  • Liposomal suspensions may also be suitable as pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. Where the compounds exhibit insufficient solubility, methods for solubilizing may be used. Such methods are known and include, but are not limited to, using cosolvents such as dimethylsulfoxide (DMSO), using surfactants such as Tween®, and dissolution in aqueous sodium bicarbonate.
  • DMSO dimethylsulfoxide
  • surfactants such as Tween®
  • Derivatives of the compounds may also be used in formulating effective pharmaceutical compositions.
  • the disclosed compounds may also be prepared with carriers that protect them against rapid elimination from the body, such as time-release formulations or coatings. Such carriers include controlled release formulations, such as, but not limited to, microencapsulated delivery systems.
  • the disclosed compounds and/or compositions can be enclosed in multiple or single dose containers.
  • the compounds and/or compositions can also be provided in kits, for example, including component parts that can be assembled for use.
  • one or more of the disclosed compounds may be provided in a lyophilized form and a suitable diluent may be provided as separated components for combination prior to use.
  • kits may include a disclosed compound and a second therapeutic agent (such as an anti-retroviral agent) for co- administration.
  • the compound and second therapeutic agent may be provided as separate component parts.
  • a kit may include a plurality of containers, each container holding one or more unit dose of the compound.
  • the containers are preferably adapted for the desired mode of administration, including, but not limited to tablets, gel capsules, sustained-release capsules, and the like for oral administration; depot products, pre-filled syringes, ampoules, vials, and the like for parenteral administration; and patches, medipads, creams, and the like for topical administration.
  • the active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the subject treated.
  • a therapeutically effective concentration may be determined empirically by testing the compounds in known in vitro and in vivo model systems for the treated disorder.
  • a therapeutically effective amount of the compound is an amount that lessens or ameliorates at least one symptom of the disorder for which the compound is administered.
  • the compositions are formulated for single dosage administration. The concentration of active compound in the drug composition will depend on absorption, inactivation, and excretion rates of the active compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art.
  • 0.1 mg to 1000 mg of a disclosed compound, a mixture of such compounds, or a physiologically acceptable salt or ester thereof, is compounded with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavor, etc., in a unit dosage form.
  • a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavor, etc. is compounded with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavor, etc.
  • the amount of active substance in those compositions or preparations is such that a suitable dosage in the range indicated is obtained.
  • unit dosage form refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.
  • the compositions are formulated in a unit dosage form, each dosage containing from about 1 mg to about 1000 mg (for example, about 2 mg to about 500 mg, about 5 mg to 50 mg, about 10 mg to 100 mg, or about 25 mg to 75 mg) of the one or more compounds.
  • the unit dosage form includes about 0.1 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, or more of the disclosed compound(s).
  • the disclosed compounds or compositions may be administered as a single dose, or may be divided into a number of smaller doses to be administered at intervals of time.
  • the therapeutic compositions can be administered in a single dose delivery, by continuous delivery over an extended time period, in a repeated administration protocol (for example, by a multi-daily, daily, weekly, or monthly repeated administration protocol). It is understood that the precise dosage, timing, and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated.
  • compositions When administered orally as a suspension, these compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners/flavoring agents.
  • these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants.
  • the compound is typically provided in a composition that protects it from the acidic environment of the stomach.
  • the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine.
  • the composition may also be formulated in combination with an antacid or other such ingredient.
  • Oral compositions will generally include an inert diluent or an edible carrier and may be compressed into tablets or enclosed in gelatin capsules.
  • the active compound or compounds can be incorporated with excipients and used in the form of tablets, capsules, or troches.
  • Pharmaceutically compatible binding agents and adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches, and the like can contain any of the following ingredients or compounds of a similar nature: a binder such as, but not limited to, gum tragacanth, acacia, corn starch, or gelatin; an excipient such as microcrystalline cellulose, starch, or lactose; a disintegrating agent such as, but not limited to, alginic acid and corn starch; a lubricant such as, but not limited to, magnesium stearate; a gildant, such as, but not limited to, colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; and a flavoring agent such as peppermint, methyl salicylate, or fruit flavoring.
  • a binder such as, but not limited to, gum tragacan
  • the dosage unit form When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil.
  • dosage unit forms can contain various other materials, which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents.
  • the compounds can also be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like.
  • a syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings, and flavors.
  • the compounds When administered orally, the compounds can be administered in usual dosage forms for oral administration.
  • dosage forms include the usual solid unit dosage forms of tablets and capsules as well as liquid dosage forms such as solutions, suspensions, and elixirs.
  • solid dosage forms When the solid dosage forms are used, it is preferred that they be of the sustained release type so that the compounds need to be administered only once or twice daily.
  • an oral dosage form is administered to the subject 1, 2, 3, 4, or more times daily.
  • the compounds can be administered orally to humans in a dosage range of 1 to 1000 mg/kg body weight in single or divided doses.
  • One illustrative dosage range is 0.1 to 200 mg/kg body weight orally (such as 0.5 to 100 mg/kg body weight orally) in single or divided doses.
  • compositions may be provided in the form of tablets containing about 1 to 1000 milligrams of the active ingredient, particularly 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, or 1000 milligrams of the active ingredient.
  • the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.
  • Injectable solutions or suspensions may also be formulated, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer’s solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
  • suitable non-toxic, parenterally-acceptable diluents or solvents such as mannitol, 1,3-butanediol, water, Ringer’s solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
  • Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include any of the following components: a sterile diluent such as water for injection, saline solution, fixed oil, a naturally occurring vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil, and the like, or a synthetic fatty vehicle such as ethyl oleate, and the like, polyethylene glycol, glycerine, propylene glycol, or other synthetic solvent; antimicrobial agents such as benzyl alcohol and methyl parabens; antioxidants such as ascorbic acid and sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates, and phosphates; and agents for the adjustment of tonicity such as sodium chloride and dextrose.
  • a sterile diluent such as water for injection, saline solution, fixed oil, a naturally occurring vegetable oil
  • Parenteral preparations can be enclosed in ampoules, disposable syringes, or multiple dose vials made of glass, plastic, or other suitable material. Buffers, preservatives, antioxidants, and the like can be incorporated as required.
  • suitable carriers include physiological saline, phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol, polypropyleneglycol, and mixtures thereof.
  • Liposomal suspensions including tissue-targeted liposomes may also be suitable as pharmaceutically acceptable carriers.
  • the compounds can be administered parenterally, for example, by IV, IM, depo-IM, SC, or depo-SC.
  • a therapeutically effective amount of about 0.1 to about 500 mg/day (such as about 1 mg/day to about 100 mg/day, or about 5 mg/day to about 50 mg/day) may be delivered.
  • the dose may be about 0.1 mg/day to about 100 mg/day, or a monthly dose of from about 3 mg to about 3000 mg.
  • the compounds can also be administered sublingually. When given sublingually, the compounds should be given one to four times daily in the amounts described above for IM administration. The compounds can also be administered intranasally. When given by this route, the appropriate dosage forms are a nasal spray or dry powder.
  • the dosage of the compounds for intranasal administration is the amount described above for IM administration.
  • these compositions may be prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents.
  • the compounds can be administered intrathecally. When given by this route, the appropriate dosage form can be a parenteral dosage form.
  • the dosage of the compounds for intrathecal administration is the amount described above for IM administration.
  • the compounds can be administered topically. When given by this route, the appropriate dosage form is a cream, ointment, or patch.
  • an illustrative dosage is from about 0.5 mg/day to about 200 mg/day. Because the amount that can be delivered by a patch is limited, two or more patches may be used.
  • the compounds can be administered rectally by suppository. When administered by suppository, an illustrative therapeutically effective amount may range from about 0.5 mg to about 500 mg.
  • these compositions may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters of polyethylene glycols, which are solid at ordinary temperatures, but liquefy and/or dissolve in the rectal cavity to release the drug.
  • a method for treating a hydrogen peroxide-present condition in a subject comprising administering a therapeutically effective amount of a compound of any one of clauses 1 to 16 to the subject in need thereof. 19.
  • the method of clause 18, wherein the hydrogen peroxide-present condition is cancer.
  • the hydrogen peroxide-present condition is a viral infection.
  • 21. The method of any one of clauses 18 to 20, wherein the compound is cleaved upon reaction with hydrogen peroxide within the subject. Examples Substrates for phosphate release were prepared through classical phosphoramidite chemistry (Scheme 1 below).
  • the substrate for phosphodiester release was prepared from 15 through sequential additions of nPrOH and allylic alcohol 16 followed by oxidation with tBuOOH to yield 17.
  • the phosphomonoester-releasing substrate 18 was prepared by swapping the order of nucleophile additions.
  • the oxidative cleavage of 17 proceeded rapidly and efficiently, with 94% of the starting material being consumed within 18 min and the phosphate 19 being produced in an equal amount within the limits of NMR detection.
  • the reaction with 18 was more complex since two cleavage events are required. Spectral overlap prohibited monitoring the formation of the phosphomonoester, though acrolein production is easily detected.
  • the reaction showed a rapid consumption of 18, with the concentration of the monocleavage product 20 increasing, then ultimately becoming negligible at 24 min. Subsequent NMR analysis confirmed that the product was the phosphate 21. Acrolein was formed in an 88% yield based on the two equivalents that are expected from the cleavage of two BAO groups.
  • the second cleavage is particularly significant since it showed that the release of a phosphate dianion is faster than protonation of the boron enolate intermediate. Additionally, the absence of a significant build-up of the monocleavage product indicates that the second cleavage is not significantly slower than the first despite proceeding through a peroxide anion adding to an anionic intermediate. As expected, these processes also showed partial hydrolysis of the pinacol ester to boronic acid intermediates prior to oxidation, though this did not impact cleavage efficiency. No phosphate release was observed in the absence of peroxide. Additional substrates were synthesized and served to highlight the scope of the process (see Table 1).
  • the capacity to deliver thiophosphates, which are substantially more stable toward enzymatic cleavage than phosphates,20 is significant, regardless of the release rate.
  • Phosphodiesters that contain an aryl and an alkyl group are accessible, as shown in the conversion of 26 to 27.
  • Diesters of this general type can be cleaved enzymatically to release monoalkyl phosphates, showing that oxidative cleavage can be merged with endogenous phosphatase processes.
  • Compound 28, in which the BPin group is replaced by the recently reported and more readily handled EPin group also releases 19 efficiently, albeit somewhat more slowly. Only 68% starting material consumption occurred after 18 minutes and complete consumption did not occur until 32 min. Notably, stability studies showed that 28 is significantly more stable toward hydrolysis compared to 17. This could be useful for applications in which the boronic acid analog does not readily traverse the cell membrane.
  • Acrolein is formed endogenously in oxidatively stressed cells through the lipid and polyamine oxidation and is ingested through environmental pollution. It is a known irritant that is also postulated to be cytotoxic due to its ability to form crosslinks with biomolecules. While conclusive IC50 values for acrolein are difficult to find, an LD10 value of 30 ⁇ M was reported in the literature.
  • the a-boryl phosphate 29 reacts with peroxide to form 19 within 12 min, indicating that this group reacts even faster than the borylallyl group.
  • This rapid breakdown contrasts the slow release of carboxylic acids from a-boryl esters that arises from coordination between the carbonyl group and boron.
  • the LD 10 for isobutyraldehyde was not reported but similar aliphatic aldehydes showed values in the range of 50 mM, indicating that by-product toxicity is not a concern for these phosphates.
  • the alcohol precursor to 30 was prepared through the borylcupration of but-2-yn-1ol and the precursor to 31 arose from Cp2Zr(H)Cl-catalyzed hydroboration of but-1-yn-3-ol.
  • the phosphates were prepared by the addition of the alcohols to dimethylphosphoryl chloride since the phosphoramidite protocol led to destruction, potentially through a [2,3]-sigmatropic rearrangement from the allylic phosphite intermediate.
  • Both of these substrates release dimethyl phosphate in the presence of H 2 O 2 , though 30 showed an additional, slower mechanism for release in the absence of peroxide that appears to proceed through an initial [3,3]-sigmatropic rearrangement based on 1 H NMR studies in CD 3 CN and buffer.
  • This pathway makes structurally similar phosphates unsuitable for biological applications.
  • the LD 10 of crotonaldehyde is 240 ⁇ M while the value for methyl vinyl ketone was not reported. This study shows that the original BAO group should be sufficient for applications in which a highly potent drug is released, the secBAO should be used for the release of moderately potent drugs, and the a-boryl ether is appropriate for releasing modestly potent drugs.
  • p-Borylbenzyl derivatives are commonly used for peroxide-mediated release studies, leading us to explore the potential for their use as oxidatively labile phosphoesters. Exposing 32 to the reaction conditions led to a more complex outcome relative to the BAO group and its derivatives. The oxidation of the boronate groups proceeded rapidly, but even after 1 h significant quantities of p-hydroxybenzyl phosphoesters remained. We conclude that the BAO groups are substantially more effective than their benzylic counterparts in phosphate release applications. Table 1.
  • the BAO groups were hydrolyzed to form boronic acid analogs (HO-Bao groups) to facilitate product isolation. The process was conducted with the initial concentration of 33 being 0.2 mM, which is an order of magnitude more dilute than the previous studies, in an effort to observe the capacity for oxidative cleavage at low substrate and oxidant concentrations.
  • AZT Nucleotide precursor A solution of i Pr 2 NPCl 2 (0.27 g, 1.3 mmol) in THF (3.5 mL) was cooled to –78 °C. A solution of boronate 6 (0.480 g, 2.61 mmol) and triethylamine (0.57 g, 5.6 mmol) in THF (7.5 mL) was added to the flask dropwise over 1 h. The reaction was slowly warmed to rt and stirred overnight. The heterogeneous mixture was filtered under N2 and the salt was washed twice with THF (3.0 mL).
  • the crude phosphoramidite was concentrated in vacuo then purified via column chromatography (14% EtOAc in cyclohexane with 1% triethylamine) to yield the desired product as a clear oil (0.315 g, 49%).
  • the phosphoramidite (0.315 g, 0.634 mmol) and azidothymidine (0.085 g, 0.32 mmol,) were dissolved in a solution of 1H-tetrazole (0.45 M in CH 3 CN, 1.40 mL, 0.634 mmol) in CH 3 CN (1.8 mL). The reaction was stirred at rt for 1 h.
  • Peroxide-release was used to initiate the enzymatic phosphorylation of AZT monophosphate 37 in the presence of ATP to form AZT diphosphate 39 and ADP (FIG.2A).
  • the F105Y mutant of thymidine monophosphate kinase (TMPK) served to promote phosphorylation in this study due to its greater tolerance of 37 relative to the wild-type enzyme.
  • This procedure (FIG. 2A) required that the TMPK mutant be expressed with an HA affinity tag in HEK293T cells, followed by purification by antibody recognition of the HA tag and immobilization on Protein A agarose beads. ATP consumption was monitored by a CellTiter-Glo assay.
  • This assay employs the conversion of luciferin to oxyluciferin to provide an ATP-dependent luminescent signal in which ATP consumption through the phosphorylation of 37 diminishes the signal.
  • ATP is stable to H 2 O 2 , 36, and a mixture of 36 and H 2 O 2 for 1 h. Prolonged exposure resulted in decomposition, even with the control study, so the assays were stopped after 1 h.
  • the assays were conducted by combining the substrate (2 ⁇ M), ATP (10 ⁇ M), and TMPK. Hydrogen peroxide (30 equiv) was added for the oxidative release studies of 36.
  • the crude reaction was purified by loading directly onto boric acid-treated silica gel (50-75% ethyl acetate in dichloromethane) to obtain the product as a brown foam (16.4 mg).
  • the foam was dissolved in acetonitrile ( ⁇ 0.75 mL) and stirred overnight at room temperature in the presence of resin-bound triphenylphosphine oxide (30.1 mg) to remove trace ruthenium.
  • the suspension was filtered and concentrated to yield the final product as a white foam (13.2 mg, 28%).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des composés qui ont une stratégie de libération de groupe phosphate sur la base de l'oxydation médiée par peroxyde de phosphoesters allyliques borylés. Des alcénylboronates peuvent être incorporés dans des phosphotriesters pour libérer des phosphodiesters et des phosphomonoesters lors de l'exposition au peroxyde d'hydrogène.
PCT/US2024/014654 2023-02-08 2024-02-06 Médicaments à base d'organophosphate clivables par oxydation Ceased WO2024167952A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202363444172P 2023-02-08 2023-02-08
US63/444,172 2023-02-08

Publications (1)

Publication Number Publication Date
WO2024167952A1 true WO2024167952A1 (fr) 2024-08-15

Family

ID=92263373

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2024/014654 Ceased WO2024167952A1 (fr) 2023-02-08 2024-02-06 Médicaments à base d'organophosphate clivables par oxydation

Country Status (1)

Country Link
WO (1) WO2024167952A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100284929A1 (en) * 2009-05-11 2010-11-11 Pinchuk Anatoly Fluorescent imaging of tumors using phospholipid ether compounds
US20120258928A1 (en) * 2008-06-11 2012-10-11 Gilead Pharmasset Llc Nucleoside cyclicphosphates
US20160220595A1 (en) * 2013-09-11 2016-08-04 Emory University Nucleotide and nucleoside therapeutic compositions and uses related thereto
US20190152926A1 (en) * 2017-10-04 2019-05-23 Japan Tobacco Inc. Nitrogen-containing heteroaryl compound and pharmaceutical use thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120258928A1 (en) * 2008-06-11 2012-10-11 Gilead Pharmasset Llc Nucleoside cyclicphosphates
US20100284929A1 (en) * 2009-05-11 2010-11-11 Pinchuk Anatoly Fluorescent imaging of tumors using phospholipid ether compounds
US20160220595A1 (en) * 2013-09-11 2016-08-04 Emory University Nucleotide and nucleoside therapeutic compositions and uses related thereto
US20190152926A1 (en) * 2017-10-04 2019-05-23 Japan Tobacco Inc. Nitrogen-containing heteroaryl compound and pharmaceutical use thereof

Similar Documents

Publication Publication Date Title
EP0533833B1 (fr) Promedicaments phosphoreux
US5627165A (en) Phosphorous prodrugs and therapeutic delivery systems using same
KR20020093824A (ko) 포스포네이트 뉴클레오티드 화합물
CN101914124B (zh) 作为抗hiv剂的核苷膦酸酯缀合物
US6156739A (en) Quinazolinone compounds
TWI630201B (zh) 化合物及其調節血紅素之用途
JP5069463B2 (ja) 抗ウイルス性ホスホネート類似物
Farrow et al. Synthesis and biological properties of novel phosphotriesters: A new approach to the introduction of biologically active nucleotides into cells
JP2016520070A (ja) リン酸／ホスホン酸誘導体及びその医薬用途
EP2891658A1 (fr) Promédicament du ténofovir et utilisations pharmaceutiques associées
US10131685B2 (en) Di- and triphosphate prodrugs
WO2006091905A1 (fr) Derives de bicyclo(3.1.0) hexane utilises comme composes antiviraux
AU2013356386A1 (en) Nucleoside kinase bypass compositions and methods
JP2018513832A (ja) ４’−チオヌクレオシドの新規な化合物、並びにその調製方法、その医薬組成物及びその用途
NZ544988A (en) Nucleobase phosphonate analogs for antiviral treatment
Egron et al. S-acyl-2-thioethyl phosphoramidate diester derivatives as mononucleotide prodrugs
WO2024167952A1 (fr) Médicaments à base d'organophosphate clivables par oxydation
KR20210038562A (ko) 젬시타빈의 인-함유 프로드러그
Villard et al. An original pronucleotide strategy for the simultaneous delivery of two bioactive drugs
Zhang et al. Synthesis, biological evaluation and molecular docking studies of Combretastatin A-4 phosphoramidates as novel anticancer prodrugs
MC1182A1 (fr) Procede de preparation de derives de la guanine
JP2001527561A (ja) アシクロビルジエステル誘導体
JPS61171498A (ja) 新規な５−フルオロ−２′−デオキシウリジン−３′−ホスフエ−ト誘導体およびその塩
EP4151646A1 (fr) Dérivés de 5-fluorouracile comme promédicaments pour le traitement du cancer
WO2025235620A1 (fr) Éthers et acétals borylés utilisés en tant que promédicaments spécifiques de cellules cancéreuses

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24753933

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE