WO2019014600A1 - Traitement d'association pour le traitement du cancer - Google Patents

Traitement d'association pour le traitement du cancer Download PDF

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WO2019014600A1
WO2019014600A1 PCT/US2018/042099 US2018042099W WO2019014600A1 WO 2019014600 A1 WO2019014600 A1 WO 2019014600A1 US 2018042099 W US2018042099 W US 2018042099W WO 2019014600 A1 WO2019014600 A1 WO 2019014600A1
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compounds
covalently
pharmaceutical composition
formula
linked conjugate
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Surya Kanta DE
Sridhar G. PRASAD
Marshall C. PETERMAN
Purnendu Kumar SHARMA
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Plex Pharmaceuticals Inc
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Plex Pharmaceuticals Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0043—Nose
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/409—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil having four such rings, e.g. porphine derivatives, bilirubin, biliverdine
    • 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
    • A61K31/4196—1,2,4-Triazoles
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0085—Brain, e.g. brain implants; Spinal cord
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers
    • A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers

Definitions

  • GBM Glioblastoma
  • TRAPl is exclusively localized in mitochondria and is crucial for the
  • Hsp90 function disrupts normal cycling of the chaperone complex, resulting in simultaneous degradation of multiple key oncogenic proteins [27]. The resulting widespread loss of these key oncogenic proteins is expected to increase the susceptibility of GBM tumors to therapeutic intervention [28].
  • inhibition of Hsp90 in animal models of GBM has shown to: (i) enhance the radiosensitization effects of PARP and PI-3 kinase inhibitor [29-31] and (ii) improve the cytotoxic effect of radiation and TMZ [32].
  • the Hsp90 inhibitor NXD30001 leads to reduction of EGFR, Akt, Cdk4 and
  • TRAPl is exclusively localized in the mitochondria and is intimately involved with cellular processes associated with apoptosis [39]. Its depletion was shown to strongly decrease GBM cell proliferation, inhibit neurosphere recovery and secondary neurosphere formation, and most importantly, enhance the therapeutic effect of TMZ in neurosphere cultures [19].
  • pharmacological targeting of TRAPl by Gamitrinibs was shown to suppress intracranial GBM growth in mice with no detectable weight loss, or organ/systemic toxicity [40].
  • Hsp90/TRAP1 inhibitor shepherdin (MW>900Da) has been shown to cause a CypD- dependent irreversible collapse of mitochondrial membrane potential in primary brain tumor cells (suggesting effective mitochondrial penetration), and to inhibit intracranial tumor growth in an animal model [20].
  • mitochondrial TRAPl inhibitor-mediated cell death and apoptosis occurs independent of antiapoptotic Bcl-2 pathways [25, 42], suggesting that TRAP1 inhibitors potentially bypasses the commonly acquired Bel -2 chemoresi stance mechanism in GBM [43].
  • formulation e.g., low bioavailability in the case of drugs that have to be localized to the brain.
  • the combination therapy requires using a chemotherapeutic agent with an Hsp90 inhibitor.
  • the chemotherapeutic agent and the Hsp90 inhibitor may be covalently linked. Therapy with chemotherapeutic agent often leads to induction of resistance caused at least in part due to overexpression of Bcl-2 family of anti-apoptotic proteins.
  • the present technology encompasses using Hsp90 inhibitor of the kind that both suppresses cancer promoting signaling pathways and inhibits TRAP1, thereby bypassing Bcl-2 mediated resistance to chemotherapeutic agents.
  • Such Hsp90 inhibitors are also referred to as dual-acting Hsp90/TRAP1 inhibitors.
  • chemotherapeutic agent such as Temozolomide
  • formulations for nasal delivery of chemotherapeutic agents and Hsp90 inhibitors make methods disclosed herein particularly suitable for treating tumors of the brain, such as Glioblastoma and related tumors.
  • formulations for nasal delivery of chemotherapeutic agents and Hsp90 inhibitors make methods disclosed herein particularly suitable for treating tumors of the brain, such as Glioblastoma and related tumors.
  • formulations for nasal delivery of chemotherapeutic agents and Hsp90 inhibitors as well as examples of dual-acting Hsp90/TRAP1 inhibitors covalently linked to a chemotherapeutic agent and their formulation for nasal delivery.
  • the present technology provides a method of treating cancer, the method comprising administering to a subject in need thereof a composition comprising an effective amount of a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor, or a pharmaceutically acceptable salt thereof.
  • TMZ covalently-linked conjugate of temolozomide
  • Hsp90 inhibitor is capable of also inhibiting TRAPl .
  • the conjugate is selected from the group consisting of (i) compounds with ganetespib as the Hsp90 inhibitor, having the formula
  • R, Ri, R 2 , R 3 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • Ri, R 2 , R 3 , R 4 and R 5 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci- Cv)cycloalkyl; halo(Ci-C7)cycloalkyl; (Ci-C7)cycloalkyl(Ci-C 4 )alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms;
  • Ri, R 2 , R 3 , and R 4 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • the method may be used to treat a variety of cancer including glioblastoma, primary malignant brain tumor, or malignant glioma.
  • the present technology provides a pharmaceutical composition for nasal administration.
  • the composition comprises a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor having any one of the formulas I to XIII; a lipid and a non-ionic surfactant; and an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the conjugate.
  • the technology includes a method of treating cancer, the method comprising administering to a subject in need thereof the composition for nasal administration.
  • composition comprising a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor having any one of the formulas I to XIII, or a pharmaceutically acceptable salt thereof, and a
  • the technology includes a method of treating cancer, the method comprising administering this composition to a subject in need thereof.
  • the present technology also includes a method of treating comprising administering to a subject in need thereof an effective amount of each of a chemotherapeutic agent and an Hsp90 inhibitor, which are not conjugated by a covalent bond.
  • the method provides a combination therapy having enhanced therapeutic effect compared to the effect of the chemotherapeutic agent or the Hsp90 inhibitor alone.
  • the Hsp90 inhibitor that are used are those capable of also inhibiting TRAP1.
  • the combination therapy produces a synergistic effect.
  • the therapy may produce an additive effect.
  • the chemotherapeutic agent may be a DNA alkylating agent, e.g., temozolomide (TMZ).
  • Hsp90 inhibitor that may be used include Genetespib, Onalespib, SNX-2112, SNX-5422, XL-888, NMS-873, NMS-E973 (or PU-H71), NVB-BEP800, Luminespib, KW-2487, Tenespimycin (17-AAG), Alvespimycin (17-DMAG), VER-49009, CH5138303, VER-50589, BIIB021, VER-155008, Geldanamycin, HSP990 (NVP-HSP990), IPI-504 (retaspimycin
  • the chemotherapeutic agent and the Hsp90 inhibitor may be administered
  • compositions simultaneously either as a single composition or as separate compositions.
  • the two may also be administered sequentially.
  • the present technology also includes a pharmaceutical composition for nasal administration.
  • the composition includes (i) a chemotherapeutic agent, (ii) an Hsp90 inhibitor, or (iii) a combination of a chemotherapeutic agent and an Hsp90 inhibitor; each combined with a lipid and a non-ionic surfactant and an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the chemotherapeutic agent and the Hsp90 inhibitor.
  • the Hsp90 inhibitors that may be used in this composition include Genetespib, Onalespib, SNX-2112, SNX-5422, XL-888, NMS-873, NMS-E973 (or PU-H71), NVB-BEP800, Luminespib, KW-2487, Tenespimycin (17-AAG), Alvespimycin (17-DMAG), VER-49009, CH5138303, VER-50589, BIIB021, VER-155008, Geldanamycin, HSP990 (NVP-HSP990), IPI-504 (retaspimycin hydrochloride), IPI-493, any of the Hsp90 inhibitors used to produce the conjugates defined by formulas I to XIII, or a compound having the formula CAP 1088 or CAP 1100 shown below
  • absorption promoting agent may a cationic polymer, e.g., chitosan.
  • the technology also includes a method of treating cancer requiring administering to a subject in need thereof a composition for nasal administration as described in this paragraph.
  • the cancer to be treated can be Glioblastoma, primary malignant brain tumor, or malignant glioma.
  • the present technology also contemplates a method of treating cancer, comprising administering to a subject in need thereof a composition comprising an effective amount of a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor, or a
  • the second Hsp90 inhibitor and the second chemotherapeutic agent may be the same as the ones used for making the covalently-linked conjugate or different.
  • the Hsp90 inhibitors that may be used in this method include
  • cancers which have developed resistance to a chemotherapeutic agent may also be treated.
  • the present technology can be further summarized by the following list of embodiments.
  • a method of treating cancer comprising administering to a subject in need thereof a composition comprising an effective amount of a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor, or a pharmaceutically acceptable salt thereof, wherein the Hsp90 inhibitor is capable of also inhibiting TRAPl, and wherein the conjugate is selected from the group consisting of
  • R, Ri, R 2 , R 3 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl; halo(Ci-Cv)cycloalkyl; (Ci-Cv)cycloalkyl(Ci-C 4 )alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms;
  • Ri, R 2 , R 3 , R 4 and R 5 are independently selected from the group consisting of hydi halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci- Cv)cycloalkyl; halo(Ci-Cv)cycloalkyl; (Ci-Cv)cycloalkyl(Ci-C4)alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms; iii) compounds with Onalespib as the Hsp90 inhibitor, having the formula
  • Ri, R 2 , R 3 , and R 4 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • halo(Ci-Cv)cycloalkyl (Ci-C7)cycloalkyl(Ci-C 4 )alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms;
  • R 6 , R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C 4 )alkoxy, (C 4 -Cv)cyclopropyl, (C2-C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • R 4 , R 5 , R5, R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C 4 )alkoxy, (C 4 -Cv)cyclopropyl, (C 2 - C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • R 4 , R 5 , R5, R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C 4 )alkoxy, (C 4 -Cv)cyclopropyl, (C 2 - C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • the compounds having formula (III) are selected from the group consisting of
  • a pharmaceutical composition for nasal administration comprising a covalently- linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor of any of embodiments 1-6; a lipid and a non-ionic surfactant; and an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the conjugate.
  • TMZ temolozomide
  • Hsp90 inhibitor of any of embodiments 1-6
  • a lipid and a non-ionic surfactant an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the conjugate.
  • composition of embodiment 9, wherein the covalently-linked conjugate is selected from one of the compounds of embodiment 6. 19. The composition of embodiment 9, wherein the absorption promoting agent is a cationic polymer.
  • composition of embodiment 19, wherein the cationic polymer is chitosan.
  • a pharmaceutical composition comprising a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor of any of embodiments 1-6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • TMZ covalently-linked conjugate of temolozomide
  • Hsp90 inhibitor of any of embodiments 1-6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • Figure 1 is a bar graph showing enhanced death of U87 human malignant glioma cells following four days of treatment with different concentrations of temozolomide (TMZ) in combination with varying concentrations of the HSP90 inhibitor Ganetespib (GTB).
  • Figure 2 is a bar graph showing enhanced death of LN229 human malignant glioma cells following four days of treatment with different concentrations of TMZ in combination with varying concentrations of the HSP90 inhibitor GTB.
  • Figure 3 is a bar graph showing enhanced death of U87 human malignant glioma cells following four days treatment of with different concentrations of TMZ in combination with varying concentrations of the HSP90 inhibitor SNX-21 12 (SNX).
  • Figure 4 is a bar graph showing enhanced death of U87 human malignant glioma cells following four days of treatment of with different concentrations of TMZ in combination with varying concentrations of the HSP90 inhibitor SNX.
  • Figure 5 is a bar graph showing enhanced death of LN229 human malignant glioma cells following six days of treatment with TMZ (50uM) in combination with the HSP90 inhibitor ⁇ (lOOnM), GTB (5 nM), or SNX (25 nM).
  • Figure 6 is a bar graph showing enhanced death of LN229 human malignant glioma cells following seven days of treatment with TMZ in combination with the HSP90 inhibitor ⁇ (lOOnM), GTB (5 nM), or SNX (25 nM).
  • Figure 7 is a bar graph demonstrating concentration dependent increase in total AKT1 expression in human malignant glioma LN229 cells upon treatment with TMZ. Inhibition of HSP90 activity by a HSP90 inhibitor (GTB or SNX) suppresses TMZ-induced AKT1 expression.
  • HSP90 inhibitor ⁇ LN229 human malignant glioma cells following seven days of treatment with TMZ in combination with the HSP90 inhibitor ⁇ (lOOnM), GTB (5 nM), or SNX (25 nM).
  • Figure 7 is a bar graph demonstrating concentration dependent increase in total AKT1 expression in human malignant glioma LN229 cells upon treatment with TMZ. Inhibition of HSP90
  • Figures 8A, 8B and 8C are bar graphs showing accumulation of dual -acting
  • FIG. 8A shows accumulation of CAP01088 in mitochondria isolated from human HEI193 neurofibromatosis type-2 (NF2) schwannoma cell lines compared with the levels in the culture medium.
  • Figure 8B shows dose-dependent decrease in tetramethylrhodamine ethyl ester (TMRE) fluorescence in HEI193 cells caused by treatment with CAPO 1088 (grey) and ganetespib (black) for 2 hours indicating
  • FIG. 8C shows mitochondrial membrane depolarization as measured by TMRE fluorescence in U251MG cells following treatment with CAPO 1100 (yellow) or CAPO 1088 (green) for 4 hours. Each bar shows mean ⁇ SEM and p value (t test).
  • Figure 9A is a bar graph showing enhanced death of LN229 human malignant glioma cells following treatment for 72 hours with various concentrations of the chemotherapeutic agent, TMZ, covalently linked to an Hsp90 inhibitor SNX-2112. Also, shown are cell death caused by the agent and the inhibitor individually at the same concentrations.
  • Figure 9B is a bar graph showing enhanced death of U87 human malignant glioma cells following treatment for 72 hours with various concentrations of the chemotherapeutic agent, TMZ, covalently linked to an Hsp90 inhibitor SNX-2112. Also, shown are cell death caused by the agent and the inhibitor individually at the same concentrations.
  • Figure 1 OA is a bar graph showing accelerated Caspase 3/7 activation mediated cell death of human U87 malignant glioma tumor cells treated with a combination of TMZ and an Hsp90 inhibitor (Ganestinib or SNX-2112).
  • Figure 10B is a bar graph showing accelerated Caspase 3/7 activation mediated cell death of LN229 malignant glioma tumor cells treated with a combination of TMZ and an Hsp90 inhibitor (Ganestinib or SNX-2112). In each case, Caspase 3/7 activity was measured after treatment with the individual agents or the combination in varying concentrations for 48 hours. DETAILED DESCRIPTION
  • the method includes administering to a patient suffering from cancer an effective amount of each of a chemotherapeutic agent and an Hsp90 inhibitor.
  • the Hsp90 inhibitor is capable of also inhibiting TRAP1.
  • the combination therapy produces an enhanced therapeutic effect compared to the effect of the chemotherapeutic agent or the Hsp90 inhibitor alone.
  • Effectiveness of current chemotherapeutic agents is often limited by the development of resistance to the agents by the cancer cells.
  • Overexpression of the Bcl-2 family of antiapoptotic proteins is one of the underlying factors for drug-induced resistance. This drug- induced resistance to driven in part by overexpression of Bcl-2 family of proteins is also an obstacle in the treatment of GBM and related cancers.
  • Hsp90 promotes cell viability with an interactome made of hundreds of client proteins, including oncogenic proteins that are the key drivers of cell proliferation, growth and survival. These client proteins are highly dependent on Hsp90 activity for folding into an active conformation. Inhibition of Hsp90 function results in simultaneous degradation of multiple key oncogenic proteins.
  • chemotherapeutic agents nearly all rely on endogenous physiological pathways for cell death in order to kill cancer cells. As such, over- activation of genes that block these pathways, e.g., genes coding for Bcl-2 family of proteins, can lead to tumor cells becoming resistant to the cytotoxic effects of the drugs. One way of overcoming this resistance is to activate cell death pathways that are not affected by the Bcl-2 family of proteins. Inhibition of TRAP 1 provides one such pathway.
  • the above-referred method of treating cancer requires use of an Hsp90 inhibitor that also inhibits TRAPl in combination with a chemotherapeutic agent.
  • the combination therapy produces a synergistic effect.
  • Chemotherapeutic agents frequently used in cancer therapy are DNA alkylating agents, and as such they may be used in the above described method.
  • Exemplary DNA alkylating agents that may be used include, but are not limited to, the following:
  • temozolomide TMZ
  • MeOS0 2 CH 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex MeOS0 2
  • Me-Lex Me-Lex
  • Cis- diamminedichloroplatinum II cis-DDP
  • mitomycin bioreductive alkylating agents cis-DDP
  • Hsp90 inhibitors that also inhibit TRAPl for use in this method include compounds having the
  • Glioblastoma is particularly suited for treatment by this method given that depletion of TRAPl leads to a marked decrease in GBM cell proliferation, inhibition of neurosphere recovery and secondary neurosphere formation, and notably, enhancement of the therapeutic effect of TMZ in neurosphere cultures [19].
  • Other brain cancers that are suitable for treatment with the above method include primary malignant brain tumor and malignant glioma.
  • the chemotherapeutic agent and the Hsp90 may be administered simultaneously in a single composition or as separate compositions. Alternatively, the chemotherapeutic agent and the Hsp90 inhibitor may be administered sequentially.
  • a pharmaceutical composition for nasal administration that includes (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, or (iii) a combination of a chemotherapeutic agent and an Hsp90 inhibitor; each with an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the chemotherapeutic agent, the Hsp90 inhibitor, or the combination.
  • Intranasal delivery is attractive as an alternative route for administration of drugs and biomolecules that are susceptible to enzymatic or acidic degradation, first-pass hepatic metabolism, and for overcoming problems of low brain bioavailability due to blood-brain barrier.
  • the composition is useful for treating cancers of many types including Glioblastoma, primary malignant brain tumor, and malignant glioma.
  • Nasally administered drugs have the advantage of access to a relatively large surface area for permeation (afforded by the large number of microvilli present in the nasal cavity), a porous endothelial membrane, and a highly-vascularized epithelium.
  • Lipid and non-ionic surfactant based elastic vesicular delivery systems which include an agent that promotes absorption to mucus membrane (e.g., a chitosan-based hybrid vesicular system), show enhanced permeability compared to conventional forms of dosing (solutions, suspensions, sprays, snuffs, emulsions, and gels).
  • These vesicular delivery systems include both stabilizing and destabilizing molecules within a single bilayer wall, which provides elasticity and also allows the system to act as a barrier modulating agent. Intranasal route has proven to be a promising means for use of these delivery systems and allows intimate and prolonged contact between the drug carrier and the mucus membrane.
  • the vesicles of the lipid and non- ionic surfactant based elastic vesicular delivery systems are highly stable and possess a high capacity for drug loading along and a controlled release profile, all of which when taken into consideration, make them a good choice as carriers for both hydrophilic and lipophilic drug molecules.
  • Absorption promoting agents contemplated to be used in the composition for nasal administration described herein include a cationic polymer, a surface active agent, a chelating agent, a mucolytic agent, a cyclodextrin, and combinations thereof.
  • cationic polymers that may be used include other polycationic carbohydrates such as but not limited to inorganic or organic salts of chitosan and modified forms of chitosan (especially more positively charged ones), polyaminoacids such as polylysine, polyquaternary compounds, protamine, polyamine, DEAE-imine, polyvinylpyridine, polythiodiethyl- aminomethylethylene (P(TDAE)), polyhistidine, DEAE-methacrylate, DEAE-acrylamide, poly-p-aminostyrene, polyox ethane, co-polymethacrylates (e.g. copolymers of HPMA, N-(2- hydroxypropyl)-methacrylamide, GAFQUAT (U
  • polyamidoamines The polycationic substances used in the technology typically have a molecular weight of 10 000 or more.
  • the chitosan (or salt thereof) preferably has an intrinsic viscosity of at least 400 ml/g, more preferably at least 500, 750 or 1000 ml/g.
  • Hsp90 inhibitor that may be used in the above described composition for nasal administration include those selected from the group consisting of Genetespib, Onalespib, SNX-2112, SNX-5422, XL-888, NMS-873, NMS-E973, or PU-H71, NVB-BEP800, Luminespib, KW-2487, Tenespimycin (17-AAG), Alvespimycin (17-DMAG), VER-49009, CH5138303, VER-50589, BIIB021, VER-155008, Geldanamycin, HSP990 (NVP-HSP990), IPI-504 (retaspimycin hydrochloride), IPI-493, and a compound having the formula
  • the Hsp90 inhibitor used in the composition is capable of also inhibiting TRAPl, i.e., they are dual-acting Hsp90/TRAP1 inhibitors.
  • exemplary Hsp90 inhibitors of this kind include Ganetespib, or a compound having the structure
  • the technology described herein further provides compounds obtained by covalently linking an Hsp90 inhibitor and a chemotherapeutic agent, e.g., TMZ, or a pharmaceutically acceptable salt thereof.
  • a chemotherapeutic agent e.g., TMZ
  • Such compounds offer certain advantages including that of a single formulation.
  • the present technology provides a method of treating cancer, the method comprising administering to a subject in need thereof a composition comprising an effective amount of a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor, or a pharmaceutically acceptable salt thereof.
  • the Hsp90 inhibitor is capable of also inhibiting TRAPl .
  • the conjugate is selected from the group consisting of
  • R, Ri, R 2 , R 3 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • Ri, R 2 , R 3 , R 4 and R 5 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci- Cv)cycloalkyl; halo(Ci-C7)cycloalkyl; (Ci-C7)cycloalkyl(Ci-C 4 )alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms;
  • Ri, R 2 , R 3 , and R 4 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • the covalently linked compounds described above may be used for preparing a pharmaceutical composition that comprises the compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
  • Such compositions, containing an effective amount of the compound may be used for treating cancer including cancers of the brain such as Glioblastoma, primary malignant brain tumor, or malignant glioma.
  • these covalently linked compounds may be included in a pharmaceutical composition for nasal administration that also includes an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the compound.
  • the composition may further include a lipid and a non-ionic surfactant.
  • absorption promoting agents including a cationic polymer such as chitosan, and such compositions may be used for treating cancer in an efficacious manner.
  • the present technology provides a method of treating cancer.
  • the method requires administering to a subject in need thereof a composition comprising an effective amount of a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor, or a pharmaceutically acceptable salt thereof.
  • TMZ covalently-linked conjugate of temolozomide
  • Hsp90 inhibitor is capable of also inhibiting TRAP1, and the conjugate is selected from the following:
  • R, Ri, R 2 , R 3 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • Ri, R 2 , R 3 , R 4 and R 5 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci- Cv)cycloalkyl; halo(Ci-C7)cycloalkyl; (Ci-C7)cycloalkyl(Ci-C 4 )alkyl; and aryl and heteroaryl optionally substituted with lower alkyl or one or more halogen atoms;
  • Ri, R 2 , R 3 , and R 4 are independently selected from the group consisting of hydrogen; halogen; (Ci-C 4 )alkyl; halo(Ci-C 4 )alkyl; (Ci-C 4 )alkoxy; halo(Ci-C 4 )alkoxy; (Ci-Cv)cycloalkyl;
  • the compounds having formula (I) are selected from the group consisting of
  • R 6 , R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C 4 )alkoxy, (C 4 -Cv)cyclopropyl, (C2-C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • the compounds having formula (II) are selected from the group consisting of
  • R 4 , R 5 , R5, R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C 4 )alkoxy, (C 4 -Cv)cyclopropyl, (C 2 - C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • the compounds having formula (III) are selected from the group consisting of
  • the compounds having formula (IV) are selected from the consisting of
  • the present technology is directed to the above six embodiments, wherein the cancer is glioblastoma, primary malignant brain tumor, or malignant glioma.
  • the present technology is directed to the above six embodiments, wherein the cancer is glioblastoma.
  • the present technology provides a pharmaceutical composition for nasal administration comprising a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor of any of the above first six embodiments; a lipid and a non-ionic surfactant; and an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the conjugate.
  • TMZ temolozomide
  • Hsp90 inhibitor of any of the above first six embodiments lipid and a non-ionic surfactant
  • an effective amount of an absorption promoting agent to allow nasal absorption of a pharmacologically effective amount of the conjugate.
  • the covalently-linked conjugate is a compound having formula (I) or (II).
  • the pharmaceutical composition of the ninth embodiment wherein the covalently-linked conjugate is a compound having formula (III).
  • the pharmaceutical composition of the ninth embodiment, wherein the covalently-linked conjugate is a compound having formula (IV) or (V).
  • the pharmaceutical composition of the ninth embodiment wherein the covalently-linked conjugate is a compound having formula selected from (VI) to (XIII).
  • the pharmaceutical composition of the ninth embodiment wherein the covalently-linked conjugate is one of the compounds of the second embodiment.
  • the pharmaceutical composition of the ninth embodiment, wherein the covalently-linked conjugate is one of the compounds of the third embodiment.
  • the pharmaceutical composition of the ninth embodiment, wherein the covalently-linked conjugate is one of the fifth embodiment.
  • the pharmaceutical composition of the ninth embodiment ⁇ wherein the absorption promoting agent is a cationic polymer.
  • the pharmaceutical composition of the ninth embodiment wherein the cationic polymer is chitosan.
  • the present technology provides a pharmaceutical composition
  • a pharmaceutical composition comprising a covalently-linked conjugate of temolozomide (TMZ) and an Hsp90 inhibitor of any of the first six embodiments, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • TMZ covalently-linked conjugate of temolozomide
  • Hsp90 inhibitor of any of the first six embodiments, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • a pharmaceutical composition of the twenty-first embodiment wherein the covalently-linked conjugate is a compound having formula (I) or (II).
  • a pharmaceutical composition of the twenty-first embodiment wherein the covalently-linked conjugate is a compound having formula (III).
  • a pharmaceutical composition of the twenty-first embodiment, wherein the covalently-linked conjugate is a compound having formula (IV) or (V).
  • a pharmaceutical composition of the twenty-first embodiment, wherein the covalently-linked conjugate is a compound having formula selected from (VI) to (XIII).
  • a twenty-second embodiment is provided a pharmaceutical composition of the twenty-first embodiment, wherein the covalently-linked conjugate is one of the third embodiment.
  • a twenty-second embodiment is provided a pharmaceutical composition of the twenty-first embodiment, wherein the covalently-linked conjugate is one of the fourth embodiment.
  • Contemplated within the scope of the present technology are all of the above described methods for treating cancer in situations where the cancer has developed resistance to a chemotherapeutic agent.
  • the terms “subject” and “patient” may be used interchangeably, and means a mammal in need of treatment, e.g., companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats and the like) and laboratory animals (e.g., rats, mice, guinea pigs and the like).
  • the subject is a human in need of treatment.
  • This technology also provides a pharmaceutical composition
  • a pharmaceutical composition comprising at least (i) an
  • Hsp90 inhibitor or (ii) a chemotherapeutic agent, or (iii) a combination of an Hsp90 inhibitor and a chemotherapeutic agent, or (iv) a compound formed by covalently linking an Hsp90 inhibitor and a chemotherapeutic agent, or a pharmaceutically-acceptable salt thereof; and a pharmaceutically-acceptable carrier.
  • pharmaceutically-acceptable carrier means a
  • composition or vehicle such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body.
  • a liquid or solid filler such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as butylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ring
  • “pharmaceuticallyacceptable salt” refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds (i.e., (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) of the present technology .
  • These salts can be prepared in situ during the final isolation and purification of the compounds of the technology , or by separately reacting a purified compound of the technology in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed.
  • Representative salts include the
  • the pharmaceutically acceptable salts of the subject compounds include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from nontoxic organic or inorganic acids.
  • such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, butionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.
  • the compounds i.e., (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) of the present technology may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically-acceptable salts with pharmaceutically-acceptable bases.
  • pharmaceutically-acceptable salts in these instances refers to the relatively non-toxic, inorganic and organic base addition salts of compounds of the present technology .
  • salts can likewise be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically- acceptable metal cation, with ammonia, or with a pharmaceutically-acceptable organic primary, secondary or tertiary amine.
  • suitable base such as the hydroxide, carbonate or bicarbonate of a pharmaceutically- acceptable metal cation, with ammonia, or with a pharmaceutically-acceptable organic primary, secondary or tertiary amine.
  • alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like.
  • Organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. See, for example, Berge et al, supra.
  • wetting agents such as sodium lauryl sulfate, magnesium stearate, and polyethylene oxide-polybutylene oxide copolymer as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.
  • Formulations of the present technology include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated and the particular mode of administration.
  • the amount of active ingredient, which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect.
  • this amount will range from about 1% to about 99% of active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.
  • Methods of preparing these formulations or compositions include the step of bringing into association a compound (i.e., (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) of the present technology with the carrier and, optionally, one or more accessory ingredients.
  • a compound i.e., (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) of the present technology with the carrier and, optionally, one or more accessory ingredients.
  • the formulations are prepared by uniformly and intimately bringing into association a compound of the present technology with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
  • Formulations of the technology suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present technology as an active ingredient.
  • a compound of the present technology may also be administered as a bolus, electuary or paste.
  • the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, and sodium starch glycolate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol,
  • compositions such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and coloring agents.
  • the pharmaceutical compositions may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
  • a tablet may be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared using binder (for example, gelatin or hydroxybutylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent.
  • Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • the tablets, and other solid dosage forms of the pharmaceutical compositions of the present technology may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxybutylmethyl cellulose in varying butortions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres.
  • compositions may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions, which can be dissolved in sterile water, or some other sterile injectable medium immediately before use.
  • These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples are embedding compositions, which can be used include polymeric substances and waxes.
  • the active ingredient can also be in micro-encapsulated form with one or more of the above-described excipients.
  • Liquid dosage forms for oral administration of the compounds of the technology include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isobutyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, butyl ene glycol, 1,3 -butyl ene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • cyclodextrins e.g., hydroxybutyl-.
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
  • Formulations of the pharmaceutical compositions of the technology for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the technology with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active pharmaceutical agents of the technology .
  • Formulations of the present technology which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be apbutriate.
  • Dosage forms for the topical or transdermal administration of a compound of this technology include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.
  • the active compound may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any preservatives, buffers, or butellants which may be required.
  • the ointments, pastes, creams and gels may contain, in addition to an active compound of this technology, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • Powders and sprays can contain, in addition to a compound of this technology, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.
  • Sprays can additionally contain customary butellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and butane.
  • Transdermal patches have the added advantage of providing controlled delivery of a compound of the present technology to the body.
  • dosage forms can be made by dissolving, or dispersing the pharmaceutical agents in the buter medium.
  • Absorption enhancers can also be used to increase the flux of the pharmaceutical agents of the technology across the skin. The rate of such flux can be controlled, by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.
  • compositions of this technology suitable for parenteral administration comprise one or more compounds of the technology in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
  • a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
  • One strategy for depot injections includes the use of polyethylene oxide copolymers wherein the vehicle is fluid at room temperature and solidifies at body temperature.
  • Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include polylactide-polyglycolide.
  • the compounds of the present technology are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably, 0.5% to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.
  • the compounds i.e., (i) a chemotherapeutic agent, (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) and pharmaceutical compositions of the present technology can be employed in combination therapies, that is, the compounds and pharmaceutical compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures.
  • the particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, the compound of the present technology may be administered concurrently with another agent for treating the same disorder), or they may achieve different effects (e.g., control of any adverse effects).
  • the compounds of the technology may be administered intravenously,
  • the compounds may be used to treat arthritic conditions in mammals (i.e., humans, livestock, and domestic animals), birds, lizards, and any other organism, which can tolerate the compounds.
  • The 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 technology.
  • a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the technology.
  • Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of
  • alkyl means a saturated straight chain or branched non- cyclic hydrocarbon having from 1 to 10 carbon atoms.
  • Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n- nonyl and n-decyl; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, iert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4- methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3- dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3
  • (Ci-C6)alkyl means a saturated straight chain or branched non-cyclic hydrocarbon having from 1 to 6 carbon atoms.
  • Representative (Ci-C 6 )alkyl groups are those shown above having from 1 to 6 carbon atoms.
  • Alkyl groups included in compounds of this invention may be optionally substituted with one or more substituents.
  • alkynyl means a saturated straight chain or branched non- cyclic hydrocarbon having from 2 to 10 carbon atoms and having at least one carbon-carbon triple bond.
  • Representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-l-butynyl, 4-pentynyl, 1- hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl, 1-octynyl, 2-octynyl, 7- octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl, 1-decynyl, 2-decynyl, 9-decynyl, 2-decy
  • alkynyl means a saturated straight chain or branched non- cyclic hydrocarbon having from 2 to 10 carbon atoms and having at least one carbon-carbon triple bond.
  • Representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-l-butynyl, 4-pentynyl, 1- hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl, 1-octynyl, 2-octynyl, 7- octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl, 1-decynyl, 2-decynyl, 9-decynyl, 2-decy
  • cycloalkyl means a saturated, mono- or polycyclic alkyl radical having from 3 to 20 carbon atoms.
  • Representative cycloalkyls include cyclopropyl, 1- methylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, -cyclodecyl, octahydro-pentalenyl, and the like.
  • Cycloalkyl groups may be optionally substituted with one or more substituents.
  • haloalkyl means and alkyl group in which one or more (including all) the hydrogen radicals are replaced by a halo group, wherein each halo group is independently selected from -F, -CI, -Br, and -I.
  • halom ethyl means a methyl in which one to three hydrogen radical(s) have been replaced by a halo group.
  • Representative haloalkyl groups include trifluorom ethyl, bromom ethyl, 1,2-dichloroethyl, 4-iodobutyl, 2- fluoropentyl, and the like.
  • alkoxy is an alkyl group which is attached to another moiety via an oxygen linker.
  • haloalkoxy is an haloalkyl group which is attached to another moiety via an oxygen linker.
  • an "aromatic ring” or “aryl” means a hydrocarbon monocyclic or polycyclic radical in which at least one ring is aromatic.
  • suitable aryl groups include, but are not limited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fused carbocyclic moieties such as 5,6,7, 8- tetrahydronaphthyl.
  • Aryl groups may be optionally substituted with one or more substituents.
  • the aryl group is a monocyclic ring, wherein the ring comprises 6 carbon atoms, referred to herein as "(C6)aryl.”
  • alkylene refers to an alkyl group that has two points of attachment.
  • (Ci-C6)alkylene refers to an alkylene group that has from one to six carbon atoms.
  • Straight chain (Ci-C 6 )alkylene groups are preferred.
  • Non-limiting examples of alkylene groups include methylene (-CH2-), ethylene (-CH2CH2-), n-propylene (-
  • Alkylene groups may be optionally substituted with one or more substituents.
  • heterocyclyl means a monocyclic (typically having 3- to 10-members) or a polycyclic (typically having 7- to 20-members) heterocyclic ring system which is either a saturated ring or a unsaturated non-aromatic ring.
  • a 3 - to 10-membered heterocycle can contain up to 5 heteroatoms; and a 7- to 20-membered heterocycle can contain up to 7 heteroatoms.
  • a heterocycle has at least on carbon atom ring member.
  • Each heteroatom is independently selected from nitrogen, which can be oxidized (e.g. , N(O)) or quaternized; oxygen; and sulfur, including sulfoxide and sulfone.
  • the heterocycle may be attached via any heteroatom or carbon atom.
  • Representative heterocycles include morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.
  • a heteroatom may be substituted with a protecting group known to those of ordinary skill in the art, for example, the hydrogen on a nitrogen may be substituted with a tert-butoxycarbonyl group.
  • the heterocyclyl may be optionally substituted with one or more substituents. Only stable isomers of such substituted heterocyclic groups are contemplated in this definition.
  • halogen or "halo" means -F, -CI, -Br or -I.
  • the term “lower” refers to a group having up to four atoms.
  • a “lower alkyl” refers to an alkyl radical having from 1 to 4 carbon atoms
  • “lower alkoxy” refers to "-0-(Ci-C4)alkyl
  • a “lower alkenyl” or “lower alkynyl” refers to an alkenyl or alkynyl radical having from 2 to 4 carbon atoms, respectively.
  • the compounds of the invention containing reactive functional groups such as (without limitation) carboxy, hydroxy, thiol, and amino moieties
  • reactive functional groups such as (without limitation) carboxy, hydroxy, thiol, and amino moieties
  • Protected derivatives are those compounds in which a reactive site or sites are blocked with one or more protecting groups.
  • suitable protecting groups for hydroxyl groups include benzyl, methoxymethyl, allyl, trimethylsilyl, tert-butyldimethylsilyl, acetate, and the like.
  • suitable amine protecting groups include benzyl oxycarbonyl, tert-butoxycarbonyl, tert-butyl, benzyl and fluorenylmethyloxy-carbonyl (Fmoc).
  • suitable thiol protecting groups include benzyl, tert-butyl, acetyl, methoxymethyl and the like.
  • Other suitable protecting groups are well known to those of ordinary skill in the art and include those found in T. W. Greene, Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein.
  • treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment.
  • treatment may be administered in the absence of symptoms.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors), i.e., prophylactic treatment. Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.
  • cancer or "tumor” are well known in the art and refer to the presence, e.g., in a subject, of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, decreased cell death/apoptosis, and certain characteristic morphological features.
  • cancer refers to all types of cancer or neoplasm or malignant tumors found in humans, including, but not limited to: leukemias, lymphomas, melanomas, carcinomas and sarcomas.
  • cancer refers to cells that have undergone a malignant transformation that makes them pathological to the host organism.
  • Primary cancer cells that is, cells obtained from near the site of malignant transformation
  • a cancer cell includes not only a primary cancer cell, but also cancer stem cells, as well as cancer progenitor cells or any cell derived from a cancer cell ancestor. This includes metastasized cancer cells, and in vitro cultures and cell lines derived from cancer cells.
  • cancers treatable according to the methods described herein include a carcinoma, sarcoma, lymphoma, melanoma, and leukemia.
  • the treatable cancers include pancreatic cancer, breast cancer, liver cancer, skin cancer, lung cancer, colon cancer, prostate cancer, thyroid cancer, bladder cancer, rectal cancer, endometrial cancer, kidney cancer, bone cancer, brain cancer (e.g. glioblastoma multiforme), cervical cancer, stomach cancer, mouth and oral cancers, neuroblastoma, testicular cancer, uterine cancer, and vulvar cancer.
  • the skin cancer is selected from the group consisting of melanoma, squamous cell carcinoma, basal cell carcinoma, and cutaneous T-cell lymphoma (CTCL).
  • CTCL cutaneous T-cell lymphoma
  • the oncological disorder is triple negative breast cancer.
  • sarcoma generally refers to a tumor which is made up of a substance like the embryonic connective tissue and is generally composed of closely packed cells embedded in a fibrillar or homogeneous substance.
  • sarcomas which can be treated with a colloidal dispersion of CoQIO in an IV formulation include, for example, a chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's
  • melanoma is taken to mean a tumor arising from the melanocytic system of the skin and other organs.
  • Melanomas which can be treated with the colloidal dispersions of CoQIO in IV formulation include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding- Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungal melanoma, and superficial spreading melanoma.
  • Carcinoma refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases.
  • Carcinomas which can be treated with compositions, as described herein, include, for example, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellul are, basaloid carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epiermoid carcinoma, carcinoma epitheliale adenoides, ex
  • any suitable anti -cancer chemotherapeutic agent can be used in either a covalent conjugates or individually in a combination in the of the methods of treating cancer described herein.
  • the chemotherapeutic agent is mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, hexamethylmelamine, busulfan, carmustine, lomustine, semustine, streptozocin, decarbazine, vincristine, vinblastine, etoposide, teniposide, paclitaxel, docetaxel, daunorubicin, idarubicin, doxorubicin, epirubicin, dactinomycin, plicamycin, mitomycin C, bleomycin, mitoxantrone, fluorouracil, floxuridine, fludarabine, mercaptopurine, thioguanine, c
  • the chemotherapeutic agent is mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, hexamethylmelamine, busulfan, carmustine, lomustine, semustine, streptozocin, decarbazine, vincristine, vinblastine, etoposide, teniposide, paclitaxel, docetaxel, daunorubicin, idarubicin,
  • doxorubicin epirubicin, dactinomycin, plicamycin, mitomycin C, bleomycin, mitoxantrone, methotrexate, fluorouracil, floxuridine, fludarabine, mercaptopurine, thioguanine, cytarabine, azacytidine, cladribine, or pentostatin.
  • the anti -cancer chemotherapeutic agent is amsacrine, azacytidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin,
  • decarbazine docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, ralitrexed, semustine, streptozocin, temozolamide, teniposide, thioguanine, thiotepa, topotecan, trimitrexate, valrubicin, vincristine, vinblastine, vindestine, or vinorelbine.
  • the anti-cancer chemotherapeutic agent is an antimetabolite.
  • it can be methotrexate, fluorouracil, floxuridine, fludarabine, mercaptopurine, thioguanine, cytarabine, azacytidine, cladribine, pentostatin, pemetrexed, raltitrexed, trimetrexate, capecitabine, or gemcitabine.
  • the technology further provides a method of treating a subject, such as a human, suffering from one of the abovementioned disorders or diseases.
  • the technology further relates to the use of provided compounds for the production of pharmaceutical compositions which are employed for the treatment and/or prophylaxis and/or amelioration of the diseases and disorders mentioned herein.
  • Glioblastoma cell lines LN229 and U87, were seeded into 96-well cell culture dishes at densities of 1,000 or 2,000 cells per well in DMEM, 10% FBS, and the antibiotics, penicillin and streptomycin.
  • the day after seeding the cells compounds (TMZ and Hsp90 inhibitors) dissolved in DMSO were added to the cell culture medium to achieve the indicated final concentrations.
  • cell viability was measured by monitoring change in absorbance upon addition of WST-8 (Dojindo) or change in fluorescence upon the addition of 20 ⁇ / ⁇ resazurin.
  • the glioblastoma cell line, LN229 was seeded into 6-well cell culture dishes at a density of 120,000 cells per well in DMEM, 10% FBS, and the antibiotics, penicillin and streptomycin. The day after seeding the cells, compounds, dissolved in DMSO, were added to the cell culture dishes to achieve the indicated final concentrations.
  • a T1 DuoSet IC ELISA kit (R&D Systems) according to the manufacturer's protocol. Total cellular protein for each treatment was measured by BCA assay (Thermo Fi sher). The proportion of AKT l to total cellular protein content in each treatment was calculated and the average and standard error from three experimental replicates were graphed. See Fig. 7
  • Glioblastoma ceil lines LN229 and U87 were seeded into 96-weli cell culture dishes at a density of 2,000 cells per well in DMEM, 10% FBS, and penicillin-streptomycin. The day after seeding the cells, compound(s) dissolved in DMSO were added to the cell culture medium to achieve the indicated final concentrations. After 48 hours of treatment with the compound(s), caspase 3/7 activity was measured by the addition of a pro-luminscent caspase 3/7 substrate, Casepase-Glo ® (Promega), according to the manufacturer's protocol. Relative fold change in caspase 3/7 activity was graphed for the average from triplicate samples. See Figs. 1 OA and 10B.
  • TRAPl inhibitor-mediated apoptosis is known to occur independent of antiapoptotic Bcl-2 pathways. Therefore, effective TRAP 1 inhibition requires penetration through mitochondrial membranes to gain access to TRAPl in the mitochondrial matrix.
  • Onalespib and temozolomide are covalently linked using Scheme 3 shown above.
  • R H, Me, Et, ⁇ - ⁇ , isopropyl, Chloro
  • Ri Me, Et, n-Pr, CHF 2 , methyl-cyclopropane, allyl, Propargyl, Cyclopropryl, cyclopentyl, cyclohexyl, etc.
  • R 2 H, Me
  • R.4, R-5, R6, R7, and R 8 are independently selected from the group consisting of hydrogen, lower alkyl, (Ci-C4)alkoxy, (C4-Cv)cyclopropyl, (C2-C 6 )alkenyl, and (C2-C 6 )alkynyl, each optionally substituted with one or more halogen atoms.
  • Temozolomide acid (compound 1) (49 mg, 0.25 mmol) was mixed with 5 mL of THF and 5 mL of DCM in a round bottom flask and was cooled to 0 °C. 4-Nitrophenol (35 mg, 0.25 mmol) was added followed by N,N-diisopropylethylamine (20 mg, 0.3 mmol) and HATU (105 mg, 0.28 mmol). The mixture was stirred at room temperature for about 5 hours. Water was added and the product was extracted with DCM (30 ml x3), the combined organic phase was washed with brine, dried over Na 2 S04, filtered and solvent was removed under reduced pressure. A pale yellow solid was obtained, which is used in the next step without further purifications.
  • the effect of covalently linked TMZ and SNX-21 12 was determined by measuring cell viability (Example 1) upon treatment of cells with the covalently linked compound.
  • Fig. 9A enhanced death of LN229 human malignant glioma cells was observed following treatment with various concentrations of the compound for 72 hours.
  • Fig. 9B enhanced death of U87 human malignant glioma cells was observed following treatment of the cells with the compound for 72 hours.
  • Example 7 Formulation of compounds for the nose to brain delivery using the chitosan- based hybrid vesicular system
  • the compounds are encapsulated in phosphatidylcholine, cholesterol, and non-ionic surfactant-based vesicles (e.g., vesicles made from Brij 76 and span 60) in varying ratios, and are hybridized with chitosan to form different suspensions. These suspensions are used to prepare various formulations using thin film hydration followed by sonication. Formulations are designed and developed using central composite design (CCD) and optimized with response surface methodology.
  • CCD central composite design
  • formulations having a combination of desirable characteristics are selected for studying transport across bovine nasal mucosa and in vitro kinetics.
  • Transport studies across bovine nasal mucosa are performed using a select few formulations as identified above. Specifically, effects of the hybrid chitosan vesicular system on drug transport across nasal epithelial layer are studied using a suitable analytical technique such as LCMS or fluorescent labeling. The ability to open cell junctions in the nasal epithelial membrane is investigated by measuring trans-epithelial electrical resistance. These studies are performed to quantify the drug permeated by the vesicles to enhance delivery across the nasal epithelium. Transport rates are compared with that obtained using active agents formulated in a buffer solution. Kinetic parameters such as passive permeation flux (Jmax), steady state permeation flux (J ss ), and lag time are evaluated in vitro across bovine nasal mucosa.
  • Jmax passive permeation flux
  • J ss steady state permeation flux
  • lag time are evaluated in vitro across bovine nasal mucosa.
  • Example 9 Investigation of in-vitro efficacy of drug formulations for delivery by intranasal route on PC-12. LN229. U87. and SHSY-5y neuronal cells PC-12, LN229, U87, and SHSY-5y neuronal cells are used as models to perform efficacy of various drug formulations comprising hybrid chitosan vesicles (see Example 7). Effects on the viability of these neuronal cells are studied to confirm potency of the formulations.
  • Example 10 In vivo evaluation for nose to brain drug delivery - microdialysis
  • vesicular system In vivo evaluation of vesicular system is be carried out in male Sprague-Dawley rats (250-300 g, Envigo, CA) under anesthesia [ketamine (80 mg/kg) and xylazine (10 mg/kg) (intraperitoneal injection)].
  • the rats are be divided into groups based on the type and/or duration of administration of the formulations: (i) solution (administered only once), (ii) solution (administered daily once for 6 days based on in vitro studies) and (iii) vesicular formulation (administered only once).
  • chitosan solution containing vesicular system of the optimized batch (at a dose of equivalent effective drug concentration) is administered via intranasal route.
  • rats chitosan solution containing vesicular system at the same dose is administered by subcutaneous injection.
  • the formulations are administered intra-nasally (subgroups 1 & 2) by placing the anesthetized rats on their back and by inserting the soft polymer capillary connected to a micro-syringe into the posterior segment of the nose. After 6 days, the rats are anesthetized and the drug concentration in the brain are sampled by micro-dialysis.
  • the micro-dialysis probes is equilibrated by perfusing KRB buffer at the rate of 2 ⁇ 7 ⁇ using a microinjection pump for a period of 1 hour with the aid of a microinjection pump. Following i.v.
  • microdialysis fractions are collected at 15, 30, 60, 90, 120, 180, 240, 300, and 360 min and quantified using LCMS/MS.
  • blood samples are collected at 15, 30, 60, 90, 120, 180, 240, 300, and 360 min from cannulated rat (jugular vein) using heparinized capillary tubes.
  • the microdialysis samples are collected and assayed for biomarkers and drug by sensitive analytical instrument like LCMS/MS and enzyme-linked immunosorbent assay.
  • Microdialysis probes (CMA 12) are calibrated in vitro by immersing the probes in dialysis media containing drug combinations
  • a syringe pump controller (BASi, West Lafayette, IN) is used to continuously perfuse the KRB buffer at a flow rate of 2.0 ⁇ / ⁇ .
  • the probes are equilibrated for a period of 1 hour and samples collected at regular intervals for a period of 6 hours. The percentage recovery is calculated as the ratio of dialysate concentration to its concentration in the dialysis medium surrounding the probe.
  • Blank CSF/plasma is obtained from untreated rats and spiked with known concentrations of drug combinations and the calibration curve plotted using data obtained from an analytical method, e.g., LCMS.
  • the rat CSF/plasma samples containing drug is extracted initially by vortexing CSF/plasma diluted with a specific solvent. Next, extraction solvent is added to the mixture and vortexed for 30 min followed by centrifugation at 4°C for a period of 10 min at lOOOg.
  • the bottom organic phase (5 mL) of the centrifuged product is collected and phosphate buffer (pH 7) added to back extract (30 min) the drug, which is then centrifuged at 4°C for a period of 10 min.
  • the separated (back extracted) phase is quantified by LCMS/MS for drug content.
  • Example I I . In vivo toxicity and efficacy of combination therapy
  • plasma and brain samples are processed at the end of the two week dosing regimens to determine the brain to plasma ratios of the administered agent.
  • one or two tolerable dosing regimens for each compound i.e., (i) a chemotherapeutic agent, or (ii) an Hsp90 inhibitor, (iii) a combination of (i) and (ii), and (iv) a compound formed by linking (i) and (ii)) with optimal client protein suppression for subsequent efficacy testing are identified.
  • mice with established orthotopic tumors are randomized into seven groups of 15 mice each and treated with placebo or one of the two treatment regimens for each combination of agents. After two weeks of treatment, five mice in each group are euthanized by paraformaldehyde perfusion and brain tumors processed for immunohistochemistry for Hsp70 and relevant client proteins (EGFR, PDGFR, Akt, and c-Met RTK) to evaluate the extent of Hsp90 suppression.
  • EGFR, PDGFR, Akt, and c-Met RTK relevant client proteins
  • mice are dosed until reaching a moribund state at which time they are euthanized, their brains processed for paraffin embedding, and then sectioned to evaluate the tumor volume.
  • combination of (i) TMZ and ganestepib and of (ii) TMZ and SNX-2112 are tested for their ability to bring about a reduction in tumor volume.
  • the doses of TMZ, ganestepib, and SNX-2112 are prepared taking into
  • Glioblastoma (cancer affecting the brain) A guide for journalists on glioblastoma and its treatment.

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Abstract

L'invention concerne une méthode de traitement du cancer comprenant l'administration d'une quantité efficace d'un agent chimiothérapeutique et d'un inhibiteur de Hsp90, soit séparément, soit sous la forme d'un conjugué covalent de ces derniers, afin de fournir un traitement d'association présentant un effet thérapeutique augmenté. L'inhibiteur de Hsp90 est également capable d'inhiber TRAP1. L'invention concerne également des compositions comprenant un agent chimiothérapeutique et un inhibiteur de Hsp90, destinées à être administrées par voie nasale.
PCT/US2018/042099 2017-07-13 2018-07-13 Traitement d'association pour le traitement du cancer Ceased WO2019014600A1 (fr)

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CN115397811A (zh) * 2020-01-20 2022-11-25 奈奥芬莱有限公司 与atp结合位点结合的异吲哚啉衍生物
CN116621812A (zh) * 2023-05-11 2023-08-22 深圳市人民医院 四氢吲唑化合物、制备方法及其在治疗食管鳞癌中的应用

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CN115397811A (zh) * 2020-01-20 2022-11-25 奈奥芬莱有限公司 与atp结合位点结合的异吲哚啉衍生物
US12517130B2 (en) 2020-01-20 2026-01-06 Neophore Limited Isoindoline derivatives which bind to an ATP binding site
WO2021245405A1 (fr) * 2020-06-01 2021-12-09 Neophore Limited Inhibiteurs de mlh1 et/ou pms2 pour le traitement du cancer
CN115996907A (zh) * 2020-06-01 2023-04-21 奈奥芬莱有限公司 用于癌症治疗的mlh1和/或pms2的抑制剂
JP2023529335A (ja) * 2020-06-01 2023-07-10 ネオフォア・リミテッド がん治療のためのｍｌｈ１および／またはｐｍｓ２の阻害剤
CN115996907B (zh) * 2020-06-01 2025-07-29 奈奥芬莱有限公司 用于癌症治疗的mlh1和/或pms2的抑制剂
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CN116621812A (zh) * 2023-05-11 2023-08-22 深圳市人民医院 四氢吲唑化合物、制备方法及其在治疗食管鳞癌中的应用
CN116621812B (zh) * 2023-05-11 2024-03-12 深圳市人民医院 四氢吲唑化合物、制备方法及其在治疗食管鳞癌中的应用

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