WO2011041731A2 - Procédé destiné à inhiber des protéines de survie apparentées aux bcl-2 - Google Patents

Procédé destiné à inhiber des protéines de survie apparentées aux bcl-2 Download PDF

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WO2011041731A2
WO2011041731A2 PCT/US2010/051201 US2010051201W WO2011041731A2 WO 2011041731 A2 WO2011041731 A2 WO 2011041731A2 US 2010051201 W US2010051201 W US 2010051201W WO 2011041731 A2 WO2011041731 A2 WO 2011041731A2
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bcl
alkyl
cells
methyl
active compound
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WO2011041731A3 (fr
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David Hockenbery
Julian Simon
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Fred Hutchinson Cancer Research Center
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Priority to US13/437,795 priority Critical patent/US8865901B2/en

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    • 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
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • 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
    • A61K31/12—Ketones
    • 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
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/137—Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4245—Oxadiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to compounds for modulating apoptosis in cells over expressing Bcl-2-related survival proteins.
  • the present invention also relates to pharmaceutical compositions containing these compounds and methods of using the compounds.
  • Bcl-2, Bcl-x L , and potentially other family members, including Mcl-1 and Bcl-w confer marked resistance to the cytotoxic effects of available anticancer agents.
  • Bcl-2 survival proteins are overexpressed in comparison to normal tissue counterparts in a significant subset of common cancers.
  • Low-basal Bcl-x L expression is a strong negative predictor of cell survival with diverse classes of chemotherapeutic agents in the sixty cell lines included in the National Cancer Institute (NCI) anticancer drug screen. For these reasons, small-molecule inhibitors of Bcl-2 and Bcl-x L are being considered as a goal for molecularly targeted cancer therapy.
  • 2-methoxy antimycin A represents the first of a novel class of inhibitors that display gain-of- function cytotoxicity, defined as enhanced killing in a cell line overexpressing a Bcl-2 survival protein target compared with an isogenic control cell line.
  • BC1-XL expression shows a positive correlation with sensitivity to 2-MeAA when comparing five cancer cell lines with the highest Bcl-x L mRNA expression in the NCI anticancer drug screen to five cell lines with lowest Bcl-x L expression, i.e., 2-MeAA is most cytotoxic against cells with the highest Bcl-x L expression.
  • Bcl-x L expression levels show a negative correlation with standard therapeutic agents, i.e., standard therapeutics are less cytotoxic against cells with high Bcl-x L expression levels. Schwartz 2007.
  • Bcl-x L Overexpression of Bcl-x L in multiple cancers correlates with resistance to chemotherapy and radiation therapy, and provides a rationale for development of small-molecule BC1-XL inhibitors. Based on knockout studies, non-neoplastic cells also require BC1-XL survival functions, particularly when challenged with cytotoxic agents.
  • One Bcl-x L inhibitor, 2-MeAA was found to be cytotoxic in cells with excess exogenous Bcl-x L but had less cytotoxicity in isogenic cell line pairs having basal levels of Bcl-x L expression (Tzung et al. Nature New Biol 2001 ;3:183-91 ; Hockenbery et al. U.S. Patent No. 7,241,804, issued July 10, 2007; and
  • Figure 1 shows EC50 values for FH279 in micromolar units as measured in the following cell lines: CFPAC-1, HPAF-II MIA PaCa, PaTu-1, PaTu-8988t, ASPC-1, FA-6, and PANC-1, as described in Example 12.
  • Figure 2 shows the cytotoxic effect of FH279 (in micromolar units) on TAMH cells overexpressing Bcl-x L (Tabx2s, squares)) compared to TAMH cells transfected with a neo expression vector (Tamh.neo, circles), as described in Example 15.
  • Figure 3 shows the relative expression levels of Bcl-2 (Fig. 3 A), Bcl-x L (Fig. 3B), and MCL-1 (Fig. 3C) measured in five pancreatic cell lines, expressed as percentage of control, maximal values, as described in Example 15.
  • Figure 4 shows the average tumor growth in an animal model following treatment with FH611 versus a vehicle control, as described in Example 17.
  • the compounds, as described herein, may be substituted with any number of substituents or functional moieties.
  • substituted whether preceded by the term “optionally” or not, and substituents contained in formulas of this invention, refer to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent.
  • the substituent may be either the same or different at every position.
  • substituted is contemplated to include all permissible substituents of organic compounds.
  • the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds.
  • heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible
  • Alkyl as used herein alone or as part of another group, refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms. In some embodiments, the alkyl employed in the invention contains 1 to 6 carbon atoms.
  • alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2- dimethylpentyl, 2,3- dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, and the like.
  • Lower alkyl as used herein, is a subset of alkyl, in some embodiments preferred, and refers to a straight or branched chain hydrocarbon group containing from 1 to 4 carbon atoms.
  • Representative examples of lower alkyl include, but are not limited to, methyl, ethyl, n- propyl, iso-propyl, n-butyl, iso-butyl,- tert- butyl, and the like.
  • alkyl or “lower alkyl” is intended to include both substituted and unsubstituted alkyl or lower alkyl unless otherwise indicated and these groups may be substituted with groups selected from halo (e.g., haloalkyl), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy (thereby creating a polyalkoxy such as polyethylene glycol), alkenyloxy, alkynyloxy, haloalkoxy, cycloalkoxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocyclolalkyloxy, mercapto, alkyl-S(0) m , haloalkyl-S(0) m , alkenyl- S(0) m , hal
  • cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano where m 0, 1, 2 or 3.
  • alkenyl refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms (or in lower alkenyl 1 to 4 carbon atoms) which include 1 to 4 double bonds in the normal chain.
  • the alkenyl employed in the invention contains 1 to 6 carbon atoms.
  • Representative examples of alkenyl include, but are not limited to, vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2,4-heptadiene, and the like.
  • alkenyl or “lower alkenyl” is intended to include both substituted and unsubstituted alkenyl or lower alkenyl unless otherwise indicated and these groups may be substituted with groups as described in connection with alkyl and lower alkyl above.
  • Alkynyl refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms (or in lower alkynyl 1 to 4 carbon atoms) which include 1 to 4 triple bond in the normal chain.
  • the alkynyl employed in the invention contain 1 to 6 carbon atoms.
  • Representative examples of alkynyl include, but are not limited to, 2-propynyl, 3-butynyl, 2- butynyl, 4-pentynyl, 3- pentynyl, and the like.
  • alkynyl or “lower alkynyl” is intended to include both substituted and unsubstituted alkynyl or lower alkynyl unless otherwise indicated and these groups may be substituted with the same groups as set forth in connection with alkyl and lower alkyl above.
  • Cycloalkyl refers to groups having 3 to 10 carbon atoms. In some embodiments, the cycloalkyl employed in the invention has 3 to 8 carbon atoms. Suitable cycloalkyls include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like, which, as in the case of other aliphatic, heteroaliphatic or hetercyclic moieties, may optionally be substituted with the same groups as set forth in connection with alkyl and lower alkyl above.
  • Heterocycloalkyl or “heterocycle”, as used herein alone or as part of another group, refers to a non-aromatic 3-, 4-, 5-, 6-, 7-, or 8- membered ring or a polycyclic group, including, but not limited to a bi- or tri-cyclic group comprising fused six-membered rings having between one and four heteroatoms independently selected from oxygen, sulfur and nitrogen, wherein (i) the nitrogen and sulfur heteroatoms may be optionally oxidized, (ii) the nitrogen heteroatom may optionally be quatemized, and (iii) may form a spiro ring or be fused with a cycloalkyl, aryl, heterocyclic ring, benzene or a
  • heterocycle employed in the invention have 3 to
  • heterocycles include, but are not limited to, l,4-dioxa-8- azaspiro[4,5]decane, morpholine, azetidine, azepine, aziridine, diazepine, 1,3-dioxolane, dioxane, dithiane, furan, imidazole, imidazoline, imidazolidine, isothiazole, isothiazoline, isothiazolidine, isoxazole, isoxazoline, isoxazolidine, morpholine, oxadiazole, oxadiazoline, oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine, piperidine, pyran, pyrazine, pyrazole, pyrazolone, pyrazolidine, pyridine, pyrimidine, pyridazine, pyrrole, pyrroline,
  • Aryl as used herein alone or as part of another group, refers to a monocyclic carbocyclic ring system or a bicyclic carbocyclic fused ring system having one or more aromatic rings. In some embodiments, the aryl employed in the invention has 3 to 14 carbon atoms.
  • aryl include, azulenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like.
  • aryl is intended to include both substituted and unsubstituted aryl unless otherwise indicated and these groups may be optionally substituted with the same groups as set forth in connection with alkyl and lower alkyl above.
  • Aryl alkyl as used herein alone or as part of another groups refers to an aryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.
  • Representative examples of aryl alkyl include, but are not limited to, benzyl, 2- phenyl ethyl, 3-phenylpropyl, 2-naphthyl-2-ethyl, and the like.
  • Heteroaryl refers to a cyclic, aromatic hydrocarbon in which one or more carbon atoms have been replaced with heteroatoms such as O, N, and S. If the heteroaryl group contains more than one heteroatom, the heteroatoms may be the same or different. In some embodiments, the heteroaryl employed in the invention have 3 to 14 carbon atoms.
  • heteroaryl groups include pyridyl, pyrimidinyl, imidazolyl, thienyl, furyl, pyrazinyl, pyrrolyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, indolyl, isoindolyl, indolizinyl, triazolyl, pyridazinyl, indazolyl, purinyl, quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, isothiazolyl, and benzo[b]thienyl.
  • pyridyl pyrimidinyl, imidazolyl, thienyl, furyl, pyrazinyl, pyrrolyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, indolyl, isoin
  • heteroaryl groups are five and six membered rings and contain from one to three heteroatoms independently selected from O, N, and S.
  • the heteroaryl group, including each heteroatom can be unsubstituted or substituted with from 1 to 4 substituents, as chemically feasible.
  • Alkoxy refers to an alkyl or lower alkyl group appended to the parent molecular moiety through an oxygen or sulfur atom.
  • the alkoxy or thioalkyl group contains 1-10 carbon atoms.
  • the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-8 carbon atoms.
  • the alkyl group contains 1-6 carbon atoms.
  • the alkyl group contains 1-4 carbon atoms.
  • alkoxy include but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert- butoxy, neopentoxy and n-hexoxy and the like.
  • thioalkyl include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.
  • Halo as used herein alone or as part of another group, refers to any suitable halogen, including -F, -CI, -Br, and -I.
  • Amin or "amino group”, as used herein alone or as part of another group, refers to the radical -NH2.
  • An “optionally substituted” amine refers to -NH2 groups wherein none, one or two of the hydrogen(s) is replaced by a suitable substituent. Disubstituted amines may have substituents that are bridging, i.e., form a heterocyclic ring structure that includes the amine nitrogen.
  • Aminoalkyl group is intended to mean the radical -NHR3, where R3 is an alkyl group.
  • Haloalkyl refers to an alkyl group having one, two, or three halogen atoms attached thereto and is exemplified by such groups as chloromethyl, bromoethyl, trifluoromethyl, and the like.
  • apoptosis refers to a regulated network of biochemical events which lead to a selective form of cell suicide, and is characterized by readily observable morphological and biochemical phenomena, such as the fragmentation of the deoxyribo-nucleic acid (DNA), condensation of the chromatin, which may or may not be associated with endonuclease activity, chromosome migration, margination in cell nuclei, the formation of apoptotic bodies, mitochondrial swelling, widening of the mitochondrial cristae, opening of the mitochondrial permeability transition pores and/or dissipation of the mitochondrial proton gradient and the like.
  • DNA deoxyribo-nucleic acid
  • condensation of the chromatin which may or may not be associated with endonuclease activity
  • chromosome migration chromosome migration
  • margination in cell nuclei the formation of apoptotic bodies
  • mitochondrial swelling widening of the mitochondrial cristae
  • apoptosis refers to at least a 5-fold greater stimulation of apoptosis, at a given concentration of an agent, including a 2-methoxy antimycin derivative, in cells that over-express a Bcl-2 family member protein as compared with cells that do not over- express the Bcl-2 family member protein (e.g., a 5-fold lower LD5 0 or IC50).
  • an agent including a 2-methoxy antimycin derivative
  • substantially non-toxic refers to an agent, including 2-MeAA, that induces apoptosis in at least about 50 percent of cells that over- express a Bcl-2 family member protein, but does not induce apoptosis in more than about 5%, more preferably less than 1%, of cells that do not over-express the Bcl-2 family member protein.
  • Bcl-2 family member protein(s) refers to an evolutionarily conserved family of proteins characterized by having one or more amino acid homology domains, BHl, BH2, BH3, and/or BH4.
  • the Bcl-2 family member proteins include Bcl-2, BC1-XL, Bcl-w, Al, Mcl-l, Bax, Bak, Bad, Bcl-xs, Bid, and the like.
  • the "Bcl-2 family member proteins” further include those proteins, or their biologically active fragments, that have at least 70%, preferably at least 80%, and more preferably at least 90% amino acid sequence identity with a Bcl-2 family member protein.
  • anti-apoptotic Bcl-2 family member protein refers to Bcl-2, BC1-XL, BCI- W, Al, Mcl-l, and other proteins characterized by having one or more amino acid homology domains, BHl, BH2, BH3, and/or BH4, and that promote cell survival by attenuating or inhibiting apoptosis.
  • the "anti-apoptotic Bcl-2 family member proteins” further include those proteins, or their biologically active fragments, that have at least 70%, preferably at least 80%», and more preferably at least 90% amino acid sequence identity with an anti- apoptotic Bcl-2 family member protein.
  • identity refers to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence, as measured using either a PILEUP or BLAST sequence comparison algorithm (see, e.g., J. Mol. Evol. 35:351-360, 1987; Higgins and Sharp, CABIOS 5: 151-153, 1989; Altschul et al, J. Mol Biol 215:403-410, 1990; Zhang et al, Nucleic Acid Res. 26:3986-3990, 1998; Altschul et al, Nucleic Acid Res. 25:3389- 33402, 1997).
  • Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith and Waterman, Adv. Appl. Math. 2:482, 1981, by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443, 1970, by the search for similarity method of Pearson and Lipman, Proc. Nat. Acad. Sci. USA 85:2444, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (see, generally, Ausubel et al, supra).
  • a mutant Bcl-2 family member amino acid sequence having one or more amino acid substitutions, additions, or deletions as compared to the wild-type protein may correspond to a second Bcl-2 family member amino acid sequence (e.g., the wild-type sequence or a functionally equivalent variant thereof) according to the convention for numbering the second Bcl-2 family member sequence, whereby the mutant sequence is aligned with the second Bcl-2 family member sequence such that at least 50%, typically at least 60%, more typically, at least 70%), preferably at least 80%), more preferably at least 90%, and even more preferably at least 95% of the amino acids in a given sequence of at least 20 consecutive amino acids are identical. Because not all positions with a given "corresponding region" need be identical, non- matching positions within a corresponding region are herein regarded as "corresponding positions.”
  • a single amino acid substitution in one ("first") mutant Bcl-2 family member protein "corresponds" to a single amino acid substitution in a second mutant Bcl-2 family member protein (e.g., Bcl-x L ) where the corresponding substituted amino acid positions of the first and second mutant proteins are identical.
  • the phrase "no substantial effect on tertiary protein structure relative to the corresponding wild- type Bcl-2 family member protein” or “no substantial alteration of tertiary protein structure relative to the corresponding wild-type Bcl-2 family member protein” means that, when a Ca trace providing a position for each Ca carbon of the mutant protein is superimposed onto a Ca trace of the corresponding wild- type protein and an a carbon root mean square (RMS) difference root mean square deviation (RJVISD) is calculated; i.e., the deviation of the mutant structure from that of the wild-type structure), the RMSD value is no more than about 1.0 A when calculated using the same structural modeling method, typically no more than about 0.75 A, even more typically no more than about 0.5 A, preferably no more than about 0.35 A, and even more preferably no more than about 0.25 A.
  • RMS carbon root mean square
  • biologically active refers to the ability of a molecule to modulate apoptosis, such as by binding to a Bcl-2 family member protein.
  • a biologically active molecule can modulate apoptosis by causing a change in the mitochondrial proton motive force gradient, by causing a change in mitochondrial swelling or the morphological
  • an effective amount or “effective” can designate an amount that maintains a desired physiological state, i.e., reduces or prevents significant decline and/or promotes improvement in the condition of interest.
  • an amount of an agent that effectively modulates the apoptotic state of an individual cell such that apoptosis is induced and/or the inappropriately regulated cell death cycle in the cell returns to a normal state.
  • the dosage will vary depending on the administration routes, symptoms and body weight of the patient but also depending upon the compound being administered.
  • terapéuticaally useful and “therapeutically effective” refer to an amount of an agent that effectively modulates the apoptotic state of an individual cell such that apoptosis is induced and/or the inappropriately regulated cell death cycle in the cell returns to a normal state.
  • diagnosis refers to an agent (e.g., an antimycin derivative) for detecting the induction or inhibition of apoptosis in a subject. These terms further include molecules useful for detecting diseases associated with apoptosis, or the susceptibility to such diseases, and for detecting over-expression or under- expression of a Bcl-2 family member protein.
  • an agent e.g., an antimycin derivative
  • over-expression and “under-expression” refer to an increase or decrease, respectively, in the levels of a Bcl-2 family member protein in a cell relative to the level of such a protein found in the same cell or a closely related non-malignant cell under normal
  • apoptosis-associated disease includes diseases, disorders, and conditions that are linked to an increased or decreased state of apoptosis in at least some of the cells of a subject.
  • diseases include neoplastic disease (e.g., cancer and other
  • proliferative diseases tumor formation, arthritis, inflammation, autoimmune disease, human immunodeficiency virus (HIV) immunodeficiency syndrome, neurodegenerative diseases, myelodysplastic syndromes (such as aplastic anemia), ischaemic syndromes (such as myocardial infarction), liver diseases which are induced by toxins (such as alcohol), alopecia, damage to the skin due to UV light, lichen planus, atrophy of the skin, cataract, and graft rejections and other premalignant and noneoplastic hyperproliferative disorders.
  • Apoptosis- associated diseases further include drug resistance associated with increased or decreased levels of an anti-apoptotic
  • a "combinatorial library” is a collection of compounds in which the compounds comprising the collection are composed of one or more types of subunits.
  • the subunits can be selected from natural or unnatural moieties, including dienes, benzene compounds, cycloalkanes, lactones, dilactones, amino acids, alkanes, and the like.
  • the compounds of the combinatorial library differ in one or more ways with respect to the number, order, type or types of
  • a combinatorial library may refer to a collection of "core molecules" which vary as to the number, type or position of R groups they contain and/or the identity of molecules composing the core molecule.
  • the collection of compounds is generated in a systematic way. Any method of systematically generating a collection of compounds differing from each other in one or more of the ways set forth above is a combinatorial library.
  • a combinatorial library can be synthesized on a solid support from one or more solid phase-bound resin starting materials.
  • the library can contain five (5) or more, preferably ten (10) or more, organic molecules, which are different from each other (i.e., five (5) different molecules and not five (5) copies of the same molecule).
  • Each of the different molecules will be present in an amount such that its presence can be determined by some means (e.g., can be isolated, analyzed, detected with a binding partner or suitable probe).
  • the actual amounts of each different molecule needed so that its presence can be determined can vary due to the actual procedures used and can change as the technologies for isolation, detection and analysis advance.
  • Preferred libraries comprise substantially equal molar amounts of each desired reaction product and do not include relatively large or small amounts of any given molecules so that the presence of such molecules dominates or is completely suppressed in any assay.
  • Combinatorial libraries are generally prepared by derivatizing a starting compound onto a solid-phase support (such as a bead).
  • the solid support has a commercially available resin attached, such as a Rink or Merrifield Resin, and the like.
  • substituents are attached to the starting compound.
  • the starting compound can comprise the dilactone moiety, or a precursor thereof.
  • Substituents are added to the starting compound, and can be varied by providing a mixture of reactants comprising the substituents. Examples of suitable substituents include, but are not limited to, the following:
  • hydrocarbon substituents i.e., aliphatic (e.g., alkyl or alkenyl), alkicyclic (e.g., cycloalkyl, cycloalkenyl) substituents, aromatic, aliphatic and alkicyclic-substituted aromatic nuclei, and the like, as well as cyclic substituents;
  • substituted hydrocarbon substituents i.e., those substituents containing nonhydrocarbon radicals which do not alter the predominantly hydrocarbon substituent; those skilled in the art will be aware of such radicals (e.g., halo (especially chloro and fluoro), alkoxy, mercapto, alkylmercapto, nitro, nitroso, sulfoxy, and the like);
  • hetero substituents that is, substituents which will, while having predominantly hydrocarbyl character, contain other than carbon atoms.
  • Suitable heteroatoms will be apparent to those of ordinary skill in the art and include, e.g., sulfur, oxygen, nitrogen, and such substituents as pyridyl, furanyl, thiophenyl, imidazolyl, and the like.
  • Heteroatoms, and typically no more than one, will be present for each carbon atom in the hydrocarbon-based substituents. Alternatively, there may be no such radicals or heteroatoms in the hydrocarbon-based substituent and it will, therefore, be purely hydrocarbon.
  • the present invention resulted from a concerted effort to create scaffolds of compounds and test them by screening for selective cytotoxicity in cells having a gain-of-function phenotype (i.e., elevated expression of a Bcl-2, anti-apoptotic family member) and minimal cytotoxicity in isogenic cells that are not overexpressing Bcl-2).
  • a gain-of-function phenotype i.e., elevated expression of a Bcl-2, anti-apoptotic family member
  • One aspect of the present invention is a method of treating an apoptosis-associated disease with the molecules resulting from this screening process.
  • One embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula I, as follows:
  • A is S, CH 2 , SO, S0 2 , CO, O, NR, and COHPh;
  • Ri is selected from the group consisting of hydroxyl and N0 2 ;
  • R 2 is H, hydroxyl, halide, or alkyl
  • R 3 is H, alkyl, or aryl linked to R4;
  • ⁇ is H, halide, alkyl, or aryl linked to R 3 ;
  • R 5 is H or alkyl.
  • the method comprises administering Formula I compound, wherein:
  • Ri is hydroxyl
  • R 2 is H, halide, methyl or t-butyl
  • R 3 is H or methyl
  • Rt is halide or methyl
  • R 5 is H, methyl or t-butyl.
  • the method comprises administering Formula I, described as follows:
  • A is selected from the group consisting of S, CH 2 , SO, and S0 2 ;
  • Ri is hydroxyl
  • R 2 is H, CI, methyl or t-butyl
  • R 3 is H or methyl
  • R 4 is H, CI, Br, or methyl
  • R 5 is H, methyl, or t-butyl.
  • the method comprises administering one or more particular embodiments of Formula I, selected from the following group:
  • the active compound of Formula I has an IC50 less than about 15
  • the active compound has an IC50 less than about 10 ⁇ in cells overexpressing BC1-XL.
  • Examples of Formula I compounds are:
  • Another embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula II, a follows:
  • R 1 is hydroxyl or N0 2 ;
  • Ri is H, alkyl, methyl halide, or COMe
  • R 2 is alkyl or halide
  • R 3 is H or hydroxyl
  • R4 is H, halide, COMe, or alkyl
  • R 5 is H or alkyl.
  • the method comprises administering Formula II, described as follows:
  • a and A 2 are each independently selected from the group consisting of H, alkyl, methyl halide, methyl alcohol, and COMe; R 2 is alkyl or halide;
  • R 3 is H or hydroxyl
  • R4 is selected from the group consisting of H, halide, COMe, and alkyl; and R 5 is H or alkyl.
  • the method comprises administering Formula II, described as follows:
  • R 2 is methyl, t-butyl, or CI
  • R 3 is hydroxyl
  • R 5 is H.
  • the method comprises administering an active compound of Formula II that has an IC50 less than about 5 ⁇ in cells overexpressing BC1-XL, and at least about 4-fold less cytotoxic in isogenic cells that are not overexpressing Bcl-x L .
  • active compound of Formula II that has an IC50 less than about 5 ⁇ in cells overexpressing BC1-XL, and at least about 4-fold less cytotoxic in isogenic cells that are not overexpressing Bcl-x L . Examples of such compounds are as follows:
  • Another embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula III, a follows:
  • R ⁇ is hydroxyl, N0 2 , or an oxadiazole formed with R 2 ;
  • R 2 is H or an oxadiazole formed with Rj;
  • R 3 is H, N0 2 , or alkyl
  • R 4 is H or halogen
  • R 5 is a substituted aryl or cycloalkyl group.
  • the method comprises administering a Formula III compounds, wherein:
  • R ⁇ is selected from the group consisting of hydroxyl, N0 2 and an oxadiazole formed with R 2 ;
  • R 2 is H or an oxadiazole formed with R ⁇
  • R 3 is H, CI, or N0 2 .
  • R 5 is 2-OH-5-Cl-Phenyl-, -CH 2 phenyl, 4-isopropyl-phenyl-, 2,4-dinitrophenyl-, and 2-chloro cyclohexane-.
  • the method comprises administering at least one of the Formula III compounds.
  • the method comprises administering an active compound of Formula III has an IC50 less than about 15 ⁇ in cells overexpressing Bcl-x L , and at least about 2-fold less cytotoxic in isogenic cells that are not overexpressing BC1-XL. More preferably, the compound has at least about 3 -fold less cytotoxic in isogenic cells that are not overexpressing BC1-XL. Examples of such compounds include the following structures:
  • Another embodiment of the invention is a method for treating an apoptosis-associated disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound of Formula IV:
  • R ⁇ and R 5 are each independently H, halide, or alkyl
  • R 2 is alkyl, or phenyl with R 3 ;
  • R 3 is H, or phenyl with R 2 ;
  • R* is H or hydroxyl
  • R 6 is H, halide, or alkyl
  • R 7 is H, or alkyl.
  • the method comprises administering a Formula IV compounds, wherein:
  • R ⁇ is H, Br, or methyl
  • R 2 is methyl, t-butyl, or phenyl with R 3 ;
  • R 3 is H, or phenyl with R 2;
  • R 5 is H, Br, CI, or methyl
  • R 6 is H, CI, Br, or t-butyl
  • R 7 is H, methyl or ethyl.
  • the method comprises administering at least one of the Formula IV compounds:
  • the method comprises administering an active compound of Formula IV that has an IC50 less than about 10 micromole/liter (10 ⁇ ) in cells overexpressing BC1-XL, and at least about 8-fold less cytotoxic in isogenic cells that are not overexpressing Bcl-x L .
  • active compound of Formula IV that has an IC50 less than about 10 micromole/liter (10 ⁇ ) in cells overexpressing BC1-XL, and at least about 8-fold less cytotoxic in isogenic cells that are not overexpressing Bcl-x L .
  • Another embodiment of the invention is a method for treating an apoptosis-associated disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound of Formula V:
  • Another embodiment of the invention is a method for treating an apoptosis-associated disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound of Formula VI:
  • A is selected from the group consisting of S, CH 2 , SO, S0 2 , CO, O, NR, or COHPh;
  • R is selected from the group consisting of hydroxyl and N0 2 ;
  • R2 is selected from the group consisting of H, alkyl, and aryl
  • R 3 is selected from the group consisting of hydrogen, OH, N0 2 , alkyl, and halogen;
  • R is selected from the group consisting of halogen, and alkyl; and R 5 is selected from the group consisting of H and alkyl.
  • Another embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula VII, a follows:
  • Ri is selected from the group consisting of H and N0 2 ;
  • R 2 is selected from the group consisting of Me and H;
  • R 3 is selected from the group consisting of Me and H.
  • Another embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula VIII, a follows:
  • Ri is selected from the group consisting of Me, Et, and cyhex;
  • R 2 is selected from the group consisting of CI, Br, Me, and H; and R 3 is selected from the group consisting of CI, Br, CI, and tBu.
  • Another embodiment of the present invention is a method for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein, such as Formula IX, as follo
  • A is S or SO
  • a 2 is CH 2 ;
  • R t is selected from the group consisting of is alkyl, S-aryl, SO-aryl, S0 2 -aryl, and CH 2 -aryl;
  • R 2 and R 4 are each independently halogen or alkyl.
  • Embodiments of the present invention further provide a pharmaceutical formulation comprising, consisting of, or consisting essentially of an active compound of Formulae I through IX as described herein in combination with a pharmaceutically acceptable carrier.
  • Embodiments of the present invention provide a method of treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an active compound as described herein.
  • an apoptosis-associated disease e.g., cancer
  • Embodiments of the present invention further provide the use of an active compound as described herein for the preparation of a medicament for treating an apoptosis-associated disease (e.g., cancer) in a subject in need thereof.
  • an apoptosis-associated disease e.g., cancer
  • the compounds of the present invention are useful for treating cells in which the cell death signal is down-regulated and the affected cell has an inappropriately diminished propensity for cell death, which is referred to herein as being in a "decreased apoptotic state. "
  • the invention further provides methods for the administration to a subject, a therapeutically effective amount of an apoptosis-modulating compound of the invention to treat an apoptosis-associated disease in which it is desirable to induce apoptosis in certain types of cells, such as virus-infected or autoantibody-expressing cells.
  • a method of treating a cancer characterized by the over- expression of a Bcl-2 family member is provided.
  • the treatment of the cancer can include the treatment of solid tumors or the treatment of leukemias.
  • the cancer can be of the skin, breast, brain, cervix, testis, and the like. More particularly, cancers may include, but are not limited to, the following organs or systems: cardiac, lung, gastrointestinal, genitourinary tract, liver, bone, nervous system, gynecological, hematologic, skin, and adrenal glands.
  • the methods herein can be used for treating gliomas (Schwannoma, glioblastoma, astrocytoma), neuroblastoma, pheochromocytoma, paraganlioma, meningioma, adrenal cortical carcinoma, kidney cancer, vascular cancer of various types, osteoblastic osteocarcoma, prostate cancer, ovarian cancer, uterine leiomyomas, salivary gland cancer, choroid plexus carcinoma, mammary cancer, pancreatic cancer, colon cancer, B and T cell lymphomas, acute and chronic myeloid or lymphoid leukemias, and multiple myeloma.
  • gliomas Rosta, glioblastoma, astrocytoma
  • neuroblastoma pheochromocytoma
  • paraganlioma paraganlioma
  • meningioma adrenal cortical carcinoma
  • kidney cancer vascular cancer of various types
  • osteoblastic osteocarcoma prostate
  • treatment may include pre-malignant conditions associated with any of the above cancers (e.g., colon adenomas, myelodysplasia syndrome).
  • methods of treating a neurodegenerative disease characterized by the over-expression of a Bcl-2 family member are provided.
  • Neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis and other diseases linked to degeneration of the brain, such as Creutzfeldt- Jakob disease and expanded polyglutamine repeat diseases.
  • Expanded polyglutamine repeat diseases with which the present invention is concerned include, but are not limited to, Huntington's disease, dentatorubral pallidoluysian atrophy, spinobulbar muscular atrophy, and spinocerebellar ataxia types 1, 2, 3, 6 and 7. See, e.g., Burke et al. US Patent No. 6,632,616.
  • methods of treating arthritis, inflammation, autoimmune diseases, human immunodeficiency virus (HIV) immunodeficiency syndrome, myelodysplastic syndromes (such as aplastic anemia), ischaemic syndromes (such as myocardial infarction), liver diseases which are induced by toxins (such as alcohol), alopecia, damage to the skin due to UV light, lichen planus, atrophy of the skin, cataract, and graft rejections are provided.
  • the compounds used in embodiments of the invention will be substantially purified prior to administration.
  • the subject can be an animal, including, but not limited to, cows, pigs, horses, chickens, cats, dogs, and the like, and is typically a mammal, and in a particular embodiment human.
  • a non-human mammal is the subject.
  • Various delivery systems are known and can be used to administer a compound of the invention, such as, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of producing the derivative, receptor-mediated endocytosis (see, e.g., Wu et al. J. Biol. Chem. 262:4429-32, 1987), and the like.
  • the apoptosis-modulating compounds are administered as therapeutic or pharmaceutical compositions by any suitable route known to the skilled artisan including, for example, intravenous, subcutaneous, intramuscular, intradermal, transdermal, intrathecal, intracerebral, intraperitoneal, intransal, epidural, and oral routes. Administration can be either rapid as by injection or over a period of time as by slow infusion or administration of slow release formulations. For treating tissues in the central nervous system, administration can be by injection or infusion into the cerebrospinal fluid (CSF). When it is intended that a compound be administered to cells in the central nervous system, administration can be with one or more other components capable of promoting penetration of the derivative across the blood- brain barrier.
  • CSF cerebrospinal fluid
  • a compound in the target tissue by any suitable route, including intravenous and intrathecal injection.
  • Pulmonary administration can also be employed, such as, for example, by use of an inhaler or nebulizer, and formulation of the compound with an aerosolizing agent.
  • the compound is coadministered with an inhibitor of esterase activity to further stabilize the compound.
  • Pharmaceutical compositions can also be administered orally in any orally acceptable dosage form including, but not limited to, capsules, tablets, caplets, lozenges, aqueous suspensions or solutions. In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch.
  • Lubricating aids such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • the agent can be combined with emulsifying and suspending aids. If desired, certain sweeteners, flavorants, or colorants can also be used.
  • the compounds of the present invention can be combined with any other tumor and/or cancer therapy.
  • the therapy can include, for example and not by way of limitation, surgery, radiation, and chemotherapy either individually or in any combination.
  • Chemotherapy can include any current known or yet to be discovered chemotherapeutic agent including but are not limited to Aceglatone; Aclarubicin; Altretamine; Aminoglutethimide; 5- Aminogleavulinic
  • Immunoglobulins Antineoplaston AIO; Asparaginase; Pegaspargase; Azacitidine; Azathioprine;
  • Carboplatin Carboquone; Carmofur; Carmustine; Chlorambucil; Chlorozotocin; Chromomycin;
  • Cisplatin Cladribine; Corynebacterium parvum; Cyclophosphamide; Cyclosporin; Cytarabine;
  • Droloxifene Echinomycin; Edatrexate; Elliptinium; Elmustine; Enloplatin; Enocitabine;
  • Fazarabine Fenretinide
  • Floxuridine Fludarabine Phosphate
  • Fluorouracil Flutamide; Formestane; Fotemustine; Gallium Nitrate; Gencitabine; Gusperimus;
  • Improsulfan Tosylate Inolimomab; lnterleukin-2; Irinotecan; JM-216; Letrozole; Lithium
  • Gamolenate Lobaplatin; Lomustine; Lonidamine; Mafosfarnide; Meiphalan; Menogaril;
  • Mitobronitol Mitoguazone Dihydrochioride; Mitolactol; Mitomycin; Mitotane; Mitozanetrone
  • Pirarubicin Piritrexim Isethionate
  • Piroxantrone Hydrochloride Plicamycin
  • porfimer Sodium porfimer Sodium
  • the compounds of the present invention can be administered locally to the area in need of treatment; this administration can be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application (e.g., in conjunction with a wound dressing after surgery), by injection, by means of a catheter, by means of a suppository, or by means of an implant, the implant being of a porous, non- porous, or gelatinous material, including membranes such as silastic membranes, or fibers.
  • administration can be by direct injection at the site (or former site) of a malignant tumor or neoplastic or pre-neoplastic tissue.
  • the compounds of the invention can be delivered in a vesicle, in particular a liposome (see, e.g., Langer, Science 249: 1527-33, 1990; Treat et al, In Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-65, 1989; Lopez-Berestein, supra, pp. 317-27).
  • a liposome see, e.g., Langer, Science 249: 1527-33, 1990; Treat et al, In Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-65, 1989; Lopez-Berestein, supra, pp. 317-27).
  • the compounds of the invention can be delivered in a controlled release system.
  • a pump can be used (see, e.g., Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201, 1987; Buchwald et ah, Surgery 88:507, 1980; Saudek et al., N. Engl. J. Med. 321 :574, 1989).
  • polymeric materials can be used (see, e.g., Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida, 1974; Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York, 1984; Ranger and Peppas, J. Macro mol. Sci. Rev. Macromol. Chem. 23:61, 1983; see also Levy et al, Science 228:190, 1985;
  • a controlled release system can be placed in proximity of the therapeutic target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, Medical
  • compositions comprise a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of the invention.
  • pharmaceutically acceptable means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more typically in humans.
  • carrier refers to a diluent, adjuvant, excipient, stabilizer, vehicle, or any combination thereof, with which the agent is formulated for administration.
  • Pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a typical carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol
  • compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release
  • compositions can be formulated as a suppository, with traditional binders and carriers such as triglycerides.
  • binders and carriers such as triglycerides.
  • composition includes an inhibiter of esterase activity as a stabilizing agent.
  • Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like.
  • suitable pharmaceutical carriers are described in, for example, Remington 's Pharmaceutical Sciences, by E.W. Martin.
  • Such compositions will contain a therapeutically effective amount of a compound of the invention, typically in purified form, together with a suitable amount of carrier so as to provide a formulation proper for administration to the subject.
  • the formulation should suit the mode of administration.
  • the compound of the present invention is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings.
  • pharmaceutical compositions for intravenous administration are solutions in sterile isotonic aqueous buffer.
  • the composition can also include a solubilizing agent and a local anesthetic to ease pain at the site of the injection.
  • the ingredients are supplied either separately or mixed together in unit dosage form. For example, as a dry lyophilized powder or water-free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent.
  • composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline.
  • an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.
  • the compounds of the invention can be formulated as neutral or salt forms.
  • pharmaceutically acceptable salt refers to a salt form of a compound permitting its use or formulation as a pharmaceutical and which retains the biological effectiveness of the free acid and base of the specified compound and that is not biologically or otherwise undesirable. Examples of such salts are described in Handbook of Pharmaceutical Salts:
  • salts include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, and the like, and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine and the like.
  • salts also include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,
  • monohydrogenphosphates dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates,
  • pharmaceutically acceptable salt includes sodium, potassium, calcium, ammonium, trialkylarylammonium and tetraalkylammonium salts.
  • prodrugs of the compounds may be used in embodiments of the invention.
  • Pharmaceutically acceptable prodrugs as used herein refers to those prodrugs of the active compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable risk/benefit ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • prodrug refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V.
  • Examples include a prodrug that is metabolized in vivo by a subject to an active drug having an activity of active compounds as described herein, wherein the prodrug is an ester of an alcohol or carboxylic acid group, if such a group is present in the compound; an acetal or ketal of an alcohol group, if such a group is present in the compound; an N-Mannich base or an imine of an amine group, if such a group is present in the compound; or a Schiff base, oxime, acetal, enol ester, oxazolidine, or thiazolidine of a carbonyl group, if such a group is present in the compound, such as described in US Patent No. 6,680,324 and US Patent No. 6,680,322.
  • the amount of the compound of the invention that is combined with the carrier to produce a single dosage form will vary, depending upon the nature of that agent and the composition of the dosage form. It should be understood, however, that a specific dosage and treatment regime for any particular patient or disease state will depend upon a variety of factors, including the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the particular disease being treated. The amount of active agent will also depend upon the specific activity of the compound and whether that agent is co-administered with any other therapeutic or prophylactic ingredients.
  • Determination of therapeutically effective dosages is typically based on animal model studies and is guided by determining effective dosages and administration protocols that significantly reduce the occurrence or severity of the apoptosis- associated disease in model subjects (e.g., in the case of treatment of malignancies, a tumor xenograft model in mice can be used.
  • a non-limiting range for a therapeutically effective amount of the compounds is about 0.001 mg/kg and about 100 mg/kg body weight per day, and in more specific embodiments between about 0.001 mg/kg and about 50 mg/kg, between about 0.01 mg kg and about 20 mg/kg. between about 0.1 and about 10 mg/kg, or between about 0.1 mg/kg and about 5 mg/kg body weight per day.
  • the invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention.
  • a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention.
  • Optionally associated with such containers can be a notice, in the form prescribed by a governmental agency, regulating the manufacture, use, or sale of pharmaceuticals or biological products. The government notice should reflect approval by the agency of manufacture, use, or sale for human administration.
  • Example 2 The effects of the compounds described in Example 2 on TAMH cells, TAMH cells overexpressing BC1-XL (TAMH-Bcl-x L ), TAMH cells transfected with Neo vector (Neo), TAMH cells over expressing a variant of Bcl-x L (A142L) is shown in Table 1 TABLE 1
  • the identification number corresponds to the entry in the NCI database for published chemical compounds.
  • the 2-OH phenyl is the most active pharmacophore.
  • the S, SO and CH2 bridges showed good selectivity.
  • triphenyl compounds appear to be more selective and more potent than the diphenyl compounds.
  • the EC50 for the compound FH279 (shown below) was determined in a panel of pancreatic cell lines.
  • the panel included CFPAC-1, HPAF-II MIA PaCa, PaTu-1, PaTu-8988t, ASPC-1, FA-6, and PANC-1 cell lines.
  • Pancreatic cancer samples were obtained from patients, with appropriate patient consent and institutional approval. Primary cells were maintained in Iscove's medium supplemented with 10% bovine calf serum, 100 ng-ml "1 stem cell factor, and 50 ng-ml "1 interleukin-3.
  • Example 1 The ability of compounds of Example 1 to induce apoptosis against solid tumors in vitro was tested.
  • OVCAR-3 an ovarian tumor
  • COLO205 a colonic tumor
  • -62 and H23 both NSCLC tumors.

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Abstract

L'invention concerne un procédé de traitement d'une maladie associée à l'apoptose chez un sujet le nécessitant, qui consiste à administrer à ce sujet une quantité thérapeutiquement efficace d'un composé actif qui est particulièrement cytotoxique contre des cellules tumorales qui surexpriment les Bcl-xL, et qui sont bien moins cytotoxiques envers les cellules isogéniques qui ne surexpriment pas les Bcl-XL. L'invention concerne également plusieurs échafaudages de composés actifs.
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KR102027532B1 (ko) * 2017-11-13 2019-10-01 연세대학교 원주산학협력단 메틸렌비스 및 관련 화합물의 자가포식 조절 기전을 이용하여 dna 토포이소머라제1 억제제의 항암 효능 증진 방법

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