WO2016111957A1 - Composés triazole de chloroquinoline, composition et utilisations - Google Patents

Composés triazole de chloroquinoline, composition et utilisations Download PDF

Info

Publication number
WO2016111957A1
WO2016111957A1 PCT/US2016/012106 US2016012106W WO2016111957A1 WO 2016111957 A1 WO2016111957 A1 WO 2016111957A1 US 2016012106 W US2016012106 W US 2016012106W WO 2016111957 A1 WO2016111957 A1 WO 2016111957A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
alkyl
optionally substituted
autophagy
cancer
Prior art date
Application number
PCT/US2016/012106
Other languages
English (en)
Inventor
Edward L. Schwartz
Lars Ulrik NORDSTROEM
Peng Wu
Original Assignee
Albert Einstein College Of Medicine, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Albert Einstein College Of Medicine, Inc. filed Critical Albert Einstein College Of Medicine, Inc.
Publication of WO2016111957A1 publication Critical patent/WO2016111957A1/fr

Links

Classifications

    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/38—Nitrogen atoms
    • C07D215/42—Nitrogen atoms attached in position 4
    • C07D215/46—Nitrogen atoms attached in position 4 with hydrocarbon radicals, substituted by nitrogen atoms, attached to said nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • Autophagy is an evolutionarily conserved, regulated catabolic process that degrades cellular proteins and organelles, allowing the recycling of their biochemical components for use in energy production and biosynthetic reactions (1-3).
  • Autophagy has a role in a number of critical cell functions, including stress response, cellular quality control, tissue homeostasis, and energy production. Increased autophagic flux was found in a large majority of human tumors, compared to non-malignant tissue, and was associated with increased tumor proliferation, invasion and metastasis, and lower patient survival (4).
  • Autophagy is also upregulated in tumor cells in response to most anti-cancer therapies, including cytotoxic chemotherapeutic drugs, targeted chemotherapeutic drugs, and radiation (1, 5-7).
  • the present disclosure provides, among other things, various useful chemical compounds that, in some embodiments, are variants of chloroquine (CQ) and/or hydroxychloroquine (HCQ).
  • CQ chloroquine
  • HCQ hydroxychloroquine
  • provided compounds show activity as autophagy inhibitors.
  • the present invention encompasses the recognition that there is an unmet need for effective inhibitors of autophagy.
  • the upregulation of autophagy has been shown to be a mechanism of drug resistance, and the use of autophagy inhibitors can reverse this effect.
  • CQ and HCQ have activity as autophagy inhibitors. CQ and HCQ also have single agent antiproliferative activity against human cancer cells. There are currently over 30 HCQ trials in cancer patients involving nearly every tumor type.
  • the present invention further encompasses the recognition that, while CQ and HCQ are effective inhibitors of autophagy in vitro, their in vivo efficacy may require concentrations at the upper range of tolerability (8, 9), and, moreover, that the low potency of CQ and HCQ may limit their efficacy in vivo.
  • the present invention also encompasses the recognition that compounds showing structural relatedness to CQ and/or HCQ have various desirable attributes including, in some embodiments, but not limited to, an ability to act as autophagy inhibitors.
  • Embodiments of this invention are directed to compounds for inhibiting autophagy in biological systems.
  • EAD1 FOG. l
  • EAD1 FOG. l
  • variants or analogs thereof may provide potent and/or effective autophagy inhibition as described herein.
  • provide compounds are variants or analogs of chloroquine or triazole.
  • a pharmaceutical composition comprises a compound according to formula I or III or as otherwise described herein in combination with a pharmaceutically acceptable carrier, additive or excipient, optionally in combination with at least one additional anticancer agent.
  • a biological system is or comprises a cell, an organ, a tissue, and/or an organism.
  • an organism is or comprises a mammal, e.g., a human.
  • an organism is or comprises a patient or subject who is suffering from or susceptible to an autophagy-related disease, disorder or condition.
  • an organism is or comprises a patient or subject who is suffering from or susceptible to cancer.
  • a composition or compound as described herein is administered to a biological system.
  • one or more effects or results of such administration may be monitored or assessed.
  • one or more effects or results on autophagy in the system is monitored or assessed.
  • administration as described herein method inhibits, treats and/or reduces the likelihood of onset of one or more symptoms of a disease, disorder or condition associated with autophagy.
  • administration as described herein method inhibits, treats and/or reduces the likelihood of onset of one or more symptoms of cancer (e.g., of metastasis).
  • the present invention provides systems (e.g., methods and/or reagents, kits, etc.) useful to identify and/or characterize analogs or variants of chloroquine and/or compounds that inhibit autophagy.
  • systems e.g., methods and/or reagents, kits, etc.
  • one or a plurality of test compounds is administered to a system and its relevant effect(s) on such system are compared with that/those of a reference compound or result achieved under comparable conditions.
  • the present disclosure provides a compound having a structure as set forth in Formula I:
  • R 1; R 2 , and R 3 are each independently H, halo (F, CI, Br or I), CN, SO2CH 3 , acyl, alkyl (in some embodiments C1-C3), alkyl halo (in some embodiments CF3), optionally substituted 0-Ci-C 6 alkyl (preferably, OCH 3 ), OCF 3 , OH, NRR' ; in some particular embodiments, R 3 is H, halo (F, CI, Br or I), CN, SO2CH 3 , acyl, alkyl (in some embodiments C1-C3), alkyl halo (in some embodiments CF3), optionally substituted 0-Ci-C 6 alkyl (preferably, OCH 3 ), OCF 3 , OH, NRR' ; in some particular embodiments, R 3 is H
  • each El is independently N or CH;
  • each E2 is independently NH, NR, O, or CRR' ;
  • each E3 is independently CH, N, O, or S;
  • R and R' are each independently H or optionally substituted Ci-Ce alkyl group
  • n 0, 1, or 2;
  • R4 is substituted or optionally substituted alkyl, substituted or optionally substituted aryl, substituted or optionally substituted aromatic, substituted or optionally substituted heteroaryl, or substituted or optionally substituted heteroaromatic; in some particular embodiments, R4 is substituted or optionally substituted benzyl; or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides a composition comprising an appropriate unit dose of a compound as described herein.
  • an appropriate unit dose is described relative to a dose of CQ or HCQ.
  • a unit dose may be an amount (or may be equivalent to CQ or HCQ in an amount) selected from 1, 4, 5, 20, 30, 40, 50, 60 , 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 2000, 5000, 10000 mg or more mg of compound).
  • a unit dose may be an amount (or may be equivalent to CQ or HCQ in an amount) appropriate for administration in a regimen that delivers or utilizes a daily dose selected from 1, 4, 5, 20, 30, 40, 50, 60 , 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 2000, 5000, or 10000 mg.
  • a provided composition may include a pharmaceutically acceptable carrier, additive or excipient.
  • the present invention provides methods comprising steps of: administering to a subject suffering from or susceptible to an autophagy associated disease disorder or condition a pharmaceutical composition as described herein.
  • the present invention provides methods of providing a compound, the methods comprising steps of:
  • the present invention provides a compound having a structure as set forth in Formula II (intermediate):
  • the present invention provides a collection of compounds (e.g., at least 100, 200, 300, 400, 500, 600, 700, 800, 1000, 5000, 10000 or more compounds), each of which shares a common structural element as set forth in Formula III:
  • the present invention provides methods of identifyi characterizing a compound having a chemical structure as set forth in Formula I:
  • FIG. 1 shows chemical structures of chloroquine, hydroxychloroquine, and EAD1.
  • FIG. 2 shows growth inhibition by chloroquine (CQ), hydroxychloroquine (HCQ), and certain indicated compounds.
  • CQ chloroquine
  • HCQ hydroxychloroquine
  • SRB sulforhodamine B
  • FIG. 3 shows growth inhibition results obtained with EAD1 in a colony growth assay study.
  • Human non-small cell lung cancer (NSCLC) cell line H460 were plated at 250 cells per well in 24 well plates. After allowing for cell attachment overnight, HCQ and EAD1 were added at the indicated concentrations. After 24 hours, drug-containing medium was removed, and drug-free medium added. Cell colonies were stained with crystal violet after an additional 10-day growth in the absence of drug. A representative experiment is shown.
  • NSCLC Human non-small cell lung cancer
  • FIG. 4 depicts induction of apoptosis by EADl, CQ and HCQ.
  • Human non-small cell lung cancer (NSCLC) cell line H460 was treated for 24 hours with the indicated concentrations of CQ, HCQ, and EADl .
  • Cells were trypsinized, stained with APC-Annexin V for 15 minutes, and analyzed by flow cytometry.
  • DAPI was used to assess cell viability. Data are means ⁇ SD of 3 experiments.
  • FIG. 5 shows that EADl increases punctate LC3 expression in lung cancer cells.
  • H3122 NSCLC cells were transfected with an LC3 -expressing vector (mCherry-EGFP- LC3B).
  • Transfected cells were treated with HCQ or EADl at the indicated concentrations for 6 hours, fixed, and analyzed by fluorescent microscopy. An increase in fluorescent puncta are seen with drug treatment.
  • the size bar is 50 ⁇ .
  • FIGS. 6a-6d illustrate an observed increase in autophagosome-associated proteins with EADl treatment.
  • H460 cells were treated for 24 hours with the indicated concentrations of HCQ and EADl (FIGS. 6a and 6c), or for the indicated times with 10 ⁇ compound (FIG. 6b).
  • FIG. 6d cells were treated with the indicated compounds (10 ⁇ for 24 hours).
  • the IC5 0 values for the inhibition of proliferation for each compound are shown.
  • Cell extracts were analyzed by immunoblots with antibodies to LC3 (FIGS. 6a, 6b and 6d) or p62 (FIG. 6c). Blots were also probed for actin.
  • FIG. 7 shows SCHEME1, which depicts an exemplary synthesis of certain chloroquinoline triazole analogs by late-stage diversification of alkyne 1.
  • FIG. 8 shows SCHEME 2, which depicts an exemplary synthetic route to certain triazole-containing chloroquine analogs.
  • FIG. 9 shows SCHEME 3, which depicts an exemplary synthetic route to certain truncated analogs.
  • Scheme 3a compound 200.
  • Scheme 3b compound 300.
  • Ri, R2, and R 3 are each independently H, halogen, CN, SO2CH 3 , acyl, alkyl, alkyl halo, optionally substituted OCi-C 6 alkyl, OCF 3 , OH, or NRR';
  • each El is independently N or CH;
  • each E2 is independently NH, NR, O, or CRR';
  • each E3 is independently CH, N, O, or S;
  • R and R' are each independently H or optionally substituted Ci-Ce alkyl
  • n 0, 1, or 2;
  • each halogen is independently F, CI, Br or I.
  • each alkyl is independently C1-C3 alkyl.
  • one or more alkyl halo is CF 3 .
  • one or more OC1-C6 alkyl is OCH 3 .
  • Ri and R2 are each independently CI or OCH 3 .
  • R3 is H.
  • R4 is optionally substituted benzyl.
  • Preferred compounds include those having the structure
  • Preferred compounds include those where R4 is C6H5, C6H 4 (4-F), CeH 4 (4-Cl), C 6 H 4 (3-C1), C 6 H 4 (2-C1), C 6 H 4 (4-Br), CH 2 C 6 H 5, CH 2 C 6 H 4 (4-C1), CH 2 C 6 H 4 (4-Br),
  • a preferred compound has the structure
  • compositions comprising one or more of any of the compounds disclosed herein, and a pharmaceutically acceptable carrier, additive or excipient.
  • the pharmaceutical composition can comprise, for example, 1 to 2000 mg of the compound.
  • Also provided are methods for treating a subject suffering from or susceptible to an autophagy associated disease, disorder or condition comprising administering to the subject one or more of the compounds or pharmaceutical compositions disclosed herein.
  • the compound or pharmaceutical composition is administered in an amount effective to ameliorate a sign or symptom of the autophagy associated disease, disorder or condition in a subject.
  • the compound or pharmaceutical composition is administered in an amount effective to inhibit autophagy in a subject.
  • the autophagy associated disease, disorder or condition can be or comprise cancer, such as, for example, a carcinoma; a sarcoma; a neuroectodermal tumor; cancer of the breast, esophagus, colon, rectum, head, kidney, liver, lung, nasopharyngeal, neck, ovary, pancreas, skin, brain, CNS, prostate, or stomach; a leukemia; a malignant lymphoma; and combinations thereof.
  • cancer such as, for example, a carcinoma; a sarcoma; a neuroectodermal tumor; cancer of the breast, esophagus, colon, rectum, head, kidney, liver, lung, nasopharyngeal, neck, ovary, pancreas, skin, brain, CNS, prostate, or stomach; a leukemia; a malignant lymphoma; and combinations thereof.
  • the methods of treatment can further comprise administering to the subject at least one additional active agent, such as, for example, an anticancer agent, such as, e.g., everolimus, trabectedin, or abraxane.
  • an anticancer agent such as, e.g., everolimus, trabectedin, or abraxane.
  • the autophagy associated disease, disorder or condition can be one or more of rheumatoid arthritis, malaria, antiphospholipid antibody syndrome, lupus, chronic urticaria and Sjogren's disease.
  • the compounds or pharmaceutical compositions disclosed herein for treating a subject suffering from or susceptible to an autophagy associated disease disorder or condition, or for treating cancer, rheumatoid arthritis, malaria, antiphospholipid antibody syndrome, lupus, chronic urticaria or Sjogren's disease are also provided.
  • the method can further comprise determining one or more characteristics or activities of the compound.
  • R is H or CH 3 .
  • additional anti-cancer agent is used to describe an additional compound which may be coadministered with one or more compounds of the present invention in the treatment of cancer.
  • agents include, for example, one or more of everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HDAC inhibitor, a c-MET inhibitor, a PAR
  • administration refers to the administration of a composition to a subject or system (e.g., to a cell, organ, tissue, organism, or relevant component or set of components thereof).
  • route of administration may vary depending, for example, on the subject or system to which the composition is being administered, the nature of the composition, the purpose of the administration, etc.
  • administration to an animal subject may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and/or vitreal.
  • administration may involve intermittent dosing.
  • administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time.
  • amelioration refers to the prevention, reduction or palliation of a state, or improvement of the state of a subject. Amelioration includes, but does not require complete recovery or complete prevention of a disease, disorder or condition (e.g., radiation injury).
  • alkyl is used herein to refer to a fully saturated monovalent radical containing carbon and hydrogen (up to 10 carbon atoms or as otherwise indicated), and which may be a straight chain, branched or cyclic.
  • alkyl groups include methyl, ethyl, n-butyl, n-heptyl, isopropyl, 2-methyl propyl, tert-butyl, neopentyl, hexyl, heptyl, octyl, nonyl, and decyl, etc.
  • an analog refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an “analog” shows significant structural similarity with the reference substance, for example sharing a core or consensus structure, but also differs in certain discrete ways.
  • an analog is a substance that can be generated from the reference substance, e.g by chemical manipulation of the reference substance. In some embodiments, an analog is a substance that can be generated through performance of a synthetic process substantially similar to (e.g., sharing a plurality of steps with) one that generates the reference substance. In some embodiments, an analog is or can be generated through performance of a synthetic process different from that used to generate the reference substance.
  • Antagonist refers to an agent that i) inhibits, decreases or reduces the effects of another agent; and/or ii) inhibits, decreases, reduces, or delays one or more biological events.
  • Antagonists may be or include agents of any chemical class including, for example, small molecules, polypeptides, nucleic acids, carbohydrates, lipids, metals, and/or any other entity that shows the relevant inhibitory activity.
  • An antagonist may be direct (in which case it exerts its influence directly upon its target) or indirect (in which case it exerts its influence by other than binding to its target; e.g., by interacting with a regulator of the target, for example so that level or activity of the target is altered).
  • the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 1 1%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).
  • aryl refers to a substituted or unsubstituted monovalent aromatic radical having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl) wherein each ring in the system contains 3 to 7 ring members.
  • Other examples include heterocyclic aromatic (heteroaromatic or heteroaryl) ring groups having one or more nitrogen, oxygen, or sulfur atoms in the ring, in particular, quinoline groups, in particular, 7- substituted-amino quinoline groups, as well as other groups.
  • Two events or entities are "associated" with one another, as that term is used herein, if the presence, level and/or form of one is correlated with that of the other.
  • a particular entity e.g., polypeptide, genetic signature, metabolite, etc.
  • two or more entities are physically "associated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another.
  • two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.
  • autophagy or "autophagocytosis” is used to describe a catabolic process in cells which involves the degradation of a cell's own components through lysosomes.
  • Autophagy is a highly regulated process of biological systems that plays a normal part in cell growth development and homeostasis helping to maintain a balance between the synthesis, degradation, and subsequent recycling of cellular products. It is a major mechanism by which a cell allocates nutrients from unnecessary processes to more-essential processes.
  • a number of autophagic processes occur in nature, all of which have the degradation of intracellular components via the lysosome as a common feature.
  • a well- known mechanism of autophagy involves the formation of a membrane around a targeted region of a cell, separating the contents from the rest of the cytoplasm. The resultant vesicle then fuses with a lysosome which subsequently degrades the contents.
  • Autophagy involves the sequestration of organelles and proteins in autophagic vesicles (AV) and degradation of this cargo through lysosomal fusion (1). Autophagy allows tumor cells to survive metabolic and therapeutic stresses (2-5). Multiple publications indicate therapy -induced autophagy is a key resistance mechanism to many anti-cancer agents.
  • bioactive agent refers to any biologically active compound or drug which may be formulated for use in the present invention.
  • exemplary bioactive agents include the compounds according to the present invention which are used to inhibit autophagy and to treat cancer as well as other compounds or agents which are otherwise described herein.
  • neoplasms include, without limitation, morphological irregularities in cells in tissue of a subject or host, as well as pathologic proliferation of cells in tissue of a subject, as compared with normal proliferation in the same type of tissue. Additionally, neoplasms include benign tumors and malignant tumors (e.g., colon tumors) that are either invasive or noninvasive.
  • Malignant neoplasms are distinguished from benign neoplasms in that the former show a greater degree of dysplasia, or loss of differentiation and orientation of cells, and have the properties of invasion and metastasis.
  • the term cancer in some embodiments, includes drug resistant cancers, including multiple drug resistant cancers.
  • neoplasms or neoplasias from which the target cell of the present invention may be derived include, without limitation, carcinomas (e.g., squamous-cell carcinomas, adenocarcinomas, hepatocellular carcinomas, and renal cell carcinomas), particularly those of the bladder, bone, bowel, breast, cervix, colon (colorectal), esophagus, head, kidney, liver, lung, nasopharyngeal, neck, ovary, pancreas, prostate, and stomach; leukemias, such as acute myelogenous leukemia, acute lymphocytic leukemia, acute promyelocytic leukemia (APL), acute T-cell lymphoblastic leukemia, adult T-cell leukemia, basophilic leukemia, eosinophilic leukemia, granulocytic leukemia, hairy cell leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic le
  • epithelial tumors including ovarian, breast, colon, head and neck, medulloblastoma and B-cell lymphoma, among others are shown to exhibit increased autophagy; in some embodiments, provided compounds are used in the treatment and/or prevention of one or more such epithelial tumors. In some embodiments, provided are used in the treatment and/or prevention of breast, colorectal, pancreatic, ovarian, lung, renal, and head & neck cancers, melanomas, glioblastomas, leukemias and/or lymphomas.
  • carriers refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered.
  • carriers can include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like.
  • carriers are or include one or more solid components.
  • a characteristic portion in the broadest sense, refers to a portion of a substance whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the substance.
  • a characteristic portion of a substance is a portion that is found in the substance and in related substances that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity.
  • a characteristic portion shares at least one functional characteristic with the intact substance.
  • a "characteristic portion" of a protein or polypeptide is one that contains a continuous stretch of amino acids, or a collection of continuous stretches of amino acids, that together are characteristic of a protein or polypeptide.
  • each such continuous stretch generally contains at least 2, 5, 10, 15, 20, 50, or more amino acids.
  • a characteristic portion of a substance e.g. , of a protein, antibody, etc.
  • a characteristic portion may be biologically active.
  • the term "combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some embodiments, two or more agents or may be administered simultaneously; in some embodiments, such agents may be administered sequentially; in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, combination therapy may be referred to as “co-administration” or "adjunct therapy.”
  • the term "comparable” is used herein to describe two (or more) sets of conditions, circumstances, individuals, or populations that are sufficiently similar to one another to permit comparison of results obtained or phenomena observed.
  • comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features.
  • sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.
  • relative language used herein e.g., enhanced, activated, reduced, inhibited, etc. will typically refer to comparisons made under comparable conditions.
  • compound is used herein to describe any specific compound or bioactive agent disclosed herein, including any and all stereoisomers (including diasteromers), individual optical isomers (enantiomers) or racemic mixtures, pharmaceutically acceptable salts and prodrug forms.
  • compound herein refers to stable compounds. Within its use in context, the term compound may refer to a single compound or a mixture of compounds as otherwise described herein. It is understood that the choice of substituents or bonds within a Markush or other group of substituents or bonds is provided to form a stable compound from those choices within that Markush or other group.
  • the term "designed" refers to an agent (i) whose structure is or was selected by the hand of man; (ii) that is produced by a process requiring the hand of man; and/or (iii) that is distinct from natural substances and other known agents.
  • determining involves manipulation of a physical sample.
  • determining involves consideration and/or manipulation of data or information, for example utilizing a computer or other processing unit adapted to perform a relevant analysis.
  • determining involves receiving relevant information and/or materials from a source.
  • determining involves comparing one or more features of a sample or entity to a comparable reference.
  • the term "dosage form" refers to physically discrete unit of a therapeutic agent for a subject (e.g., a human patient) to be treated.
  • Each unit contains a predetermined quantity of active material calculated or demonstrated to produce a desired therapeutic effect when administered to a relevant population according to an appropriate dosing regimen.
  • such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen).
  • a medical professional e.g., a medical doctor
  • dosing regimen or “therapeutic regimen” is a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time.
  • a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.
  • a dosing regimen comprises a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regime comprises a plurality of doses and at least two different time periods separating individual doses.
  • the therapeutic agent is administered continuously (e.g., by infusion) over a predetermined period.
  • a therapeutic agent is administered once a day (QD) or twice a day (BID).
  • a dosing regimen comprises a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount.
  • a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount
  • a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).
  • excipient refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example to provide or contribute to a desired consistency or stabilizing effect.
  • suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.
  • inhibitor refers to the partial or complete elimination of a potential effect, while inhibitors are compounds that have the ability to inhibit.
  • compositions including a chemical entity that can exist in a variety of isomeric forms include a plurality of such forms; in some embodiments such compositions include only a single form.
  • compositions including a chemical entity that can exist as a variety of optical isomers include a racemic population of such optical isomers; in some embodiments such compositions include only a single optical isomer and/or include a plurality of optical isomers that together retain optical activity.
  • the term "patient” or “subject” refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, farm animals and/or humans). In some embodiments, a patient is a human. A human includes pre- and post-natal forms. In some embodiments, a patient is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient displays one or more symptoms of a disorder or condition. In some embodiments, a patient has been diagnosed with one or more disorders or conditions.
  • animals e.g., mammals such as mice, rats, rabbits, non-human primates, farm animals and/or humans.
  • a patient is a human.
  • a human includes pre- and post-natal forms.
  • a patient is suffering from or susceptible to one or more disorders or conditions.
  • a patient displays one or more symptoms of a
  • composition refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers.
  • active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.
  • compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; trans dermally; or nasally, pulmonary, and/or to other mucosal surfaces.
  • oral administration for example, drenches (aqueous or non-aqueous
  • pharmaceutically acceptable refers to agents that, within the scope of sound medical judgment, are suitable for use in contact with tissues of human beings and/or animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • the term "pharmaceutically acceptable salt” refers to those salts 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, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al, describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
  • Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases.
  • Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate
  • physiological conditions has its art-understood meaning referencing conditions under which cells or organisms live and/or reproduce. In some embodiments, the term refers to conditions of the external or internal mileu that may occur in nature for an organism or cell system.
  • physiological conditions are those conditions present within the body of a human or non-human animal, especially those conditions present at and/or within a surgical site.
  • Physiological conditions typically include, e.g., a temperature range of 20 - 40°C, atmospheric pressure of 1 , pH of 6- 8, glucose concentration of 1-20 mM, oxygen concentration at atmospheric levels, and gravity as it is encountered on earth.
  • conditions in a laboratory are manipulated and/or maintained at physiologic conditions.
  • physiological conditions are encountered in an organism.
  • prevention refers to a delay of onset, and/or reduction in frequency and/or severity of one or more symptoms of a particular disease, disorder or condition. In some embodiments, prevention is assessed on a population basis such that an agent is considered to "prevent” a particular disease, disorder or condition if a statistically significant decrease in the development, frequency, and/or intensity of one or more symptoms of the disease, disorder or condition is observed in a population susceptible to the disease, disorder, or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.
  • Radiotherapy or “radiation therapy” is used to describe therapy for cancer which may be used in conjunction with the present compounds.
  • Radiation therapy uses high doses of radiation, such as X-rays, or other energy sources such as radioisotopes (gamma, beta or alpha emitters), to destroy cancer cells.
  • the radiation damages the genetic material of the cells so that they cannot grow.
  • Radioisotopes gamma, beta or alpha emitters
  • Radiation therapy may be used in combination with the presently claimed compounds, alone or in combination with additional anticancer compounds as otherwise disclosed herein, depending on the cancer to be treated.
  • Radiotherapy therapy is most effective in treating cancers that have not spread outside the area of the original cancer, but it also may be used if the cancer has spread to nearby tissue. Radiotherapy is sometimes used after surgery to destroy any remaining cancer cells and to relieve pain from metastatic cancer.
  • a reference agent describes a standard or control agent, animal, individual, population, sample, sequence or value against which an agent, animal, individual, population, sample, sequence or value of interest is compared.
  • a reference agent, animal, individual, population, sample, sequence or value is tested and/or determined substantially simultaneously with the testing or determination of the agent, animal, individual, population, sample, sequence or value of interest.
  • a reference agent, animal, individual, population, sample, sequence or value is a historical reference, optionally embodied in a tangible medium.
  • a reference agent, animal, individual, population, sample, sequence or value is determined or characterized under conditions comparable to those utilized to determine or characterize the agent, animal, individual, population, sample, sequence or value of interest.
  • sample typically refers to a biological sample obtained or derived from a source of interest, as described herein.
  • a source of interest comprises an organism, such as an animal or human.
  • a biological sample is or comprises biological tissue or fluid.
  • a biological sample may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or broncheoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and/or excretions; and/or cells therefrom, etc.
  • a biological sample is or comprises cells obtained from an individual.
  • obtained cells are or include cells from an individual from whom the sample is obtained.
  • a sample is a "primary sample" obtained directly from a source of interest by any appropriate means.
  • a primary biological sample is obtained by methods selected from the group consisting of biopsy (e.g. , fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e.g., blood, lymph, feces etc.), etc.
  • sample refers to a preparation that is obtained by processing (e.g., by removing one or more components of and/or by adding one or more agents to) a primary sample. For example, filtering using a semi-permeable membrane.
  • processing e.g., by removing one or more components of and/or by adding one or more agents to
  • a primary sample For example, filtering using a semi-permeable membrane.
  • Such a “processed sample” may comprise, for example nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components, etc.
  • small molecule means a low molecular weight organic and/or inorganic compound.
  • a "small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size. In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2000 g/mol, less than about 1500 g/mol, less than about 1000 g/mol, less than about 800 g/mol, or less than about 500 g/mol. In some embodiments, a small molecule is not a polymer.
  • a small molecule does not include a polymeric moiety.
  • a small molecule is not a protein or polypeptide (e.g., is not an oligopeptide or peptide).
  • a small molecule is not a polynucleotide (e.g., is not an oligonucleotide).
  • a small molecule is not a polysaccharide.
  • a small molecule does not comprise a polysaccharide (e.g., is not a glycoprotein, proteoglycan, glycolipid, etc.).
  • a small molecule is not a lipid.
  • a small molecule is biologically active.
  • a small molecule is detectable (e.g., comprises at least one detectable moiety).
  • a small molecule is a therapeutic.
  • the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest.
  • One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result.
  • the term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.
  • substituted as that term relates to alkyl groups which are described above include one or more functional groups such as lower alkyl groups containing 1-6 carbon atoms which are optionally substituted with 1 or 2 hydroxyl groups or between 1 and 5 (preferably 3-5) fluoro groups, acyl (Ci-Ce), halogen (F, CI, Br, I, e.g., alkyl halos, e.g., CF3), amido, hydroxyl, carboxy/carboxylic acid, thioamido, cyano, nitro, alkenyl (C2-C6) alkynyl (C 2 -C 6 ), azido, alkoxy (Ci-Ce), (including alkoxy groups which are further optionally substituted with a C1-C6 alkoxy group thus producing a diether group), amino, C1-C6 alkylamino and dialkyl-amino, where the alkyl groups may be optionally substituted
  • Preferred substituents on alkyl groups include, for example, at least one hydroxyl group, an amine, monoalkyl amine or dialkyl amine (where one or both alkyl groups is itself further optionally substituted with a dialkyl amine or an amine optionally substituted with one or two (preferably one) 7-substituted-4-quinolinyl group(s) where the amine group is bonded to the 4-position of the quinolinyl group) or an alkoxy group (e.g. methoxy or ethoxy) which may be further substituted with an alkoxy group, preferably a methoxy group, thus forming a diether substituent.
  • an alkoxy group e.g. methoxy or ethoxy
  • substituted as used in the term “substituted aryl, substituted aromatic, substituted heteroaryl, or substituted heteroaromatic” herein signifies, for example, that a substitution on the 7-position of 4-aminoquinoline may be present, said substituents being selected from atoms and groups, which when present enhance the activity of the compound as an inhibitor of autophagy.
  • substituents that may be present in a substituted aromatic or heteroaromatic group include, but are not limited to, groups such as H, halo (F, CI, Br or I), CN, NO2, optionally substituted C1-C6 alkyl (when substituted, preferably substituted with 1 or 2 hydroxyl groups or 3-5 fluoro groups), optionally substituted O-C1-C6 alkyl (preferably, OCH 3 ), optionally substituted C2-C7 acyl (preferably acetyl) or optionally substituted C2-C7 ester (oxycarbonyl ester or carboxyester, preferably carboxy ester). It is noted that each of the substituents disclosed herein may themselves be substituted.
  • An individual who is "susceptible to" a disease, disorder, or condition is at risk for developing the disease, disorder, or condition.
  • an individual who is susceptible to a disease, disorder, or condition does not display any symptoms of the disease, disorder, or condition.
  • an individual who is susceptible to a disease, disorder, or condition has not been diagnosed with the disease, disorder, and/or condition.
  • an individual who is susceptible to a disease, disorder, or condition is an individual who has been exposed to conditions associated with development of the disease, disorder, or condition.
  • a risk of developing a disease, disorder, and/or condition is a population-based risk (e.g., family members of individuals suffering from the disease, disorder, or condition).
  • symptoms are reduced" when one or more symptoms of a particular disease, disorder or condition is reduced in magnitude (e.g. , intensity, severity, etc.) and/or frequency.
  • magnitude e.g. , intensity, severity, etc.
  • frequency e.g., frequency of a particular symptom.
  • the phrase "therapeutic agent” in general refers to any agent that elicits a desired pharmacological effect when administered to an organism.
  • an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across an appropriate population.
  • the appropriate population may be a population of model organisms.
  • an appropriate population may be defined by various criteria, such as a certain age group, gender, genetic background, preexisting clinical conditions, etc.
  • a therapeutic agent is a substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
  • a “therapeutic agent” is an agent that has been or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a “therapeutic agent” is an agent for which a medical prescription is required for administration to humans.
  • a "therapeutic regimen”, as that term is used herein, refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome.
  • a "therapeutically effective amount” is meant an amount that produces the desired effect for which it is administered.
  • the term refers to an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and/or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and/or condition.
  • a therapeutically effective amount is one that reduces the incidence and/or severity of, and/or delays onset of, one or more symptoms of the disease, disorder, and/or condition.
  • a therapeutically effective amount does not in fact require successful treatment be achieved in a particular individual.
  • a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment.
  • reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.).
  • tissue e.g., a tissue affected by the disease, disorder or condition
  • fluids e.g., blood, saliva, serum, sweat, tears, urine, etc.
  • a therapeutically effective amount of a particular agent or therapy may be formulated and/or administered in a single dose.
  • a therapeutically effective agent may be formulated and/or administered in a plurality of doses, for example, as part of a dosing regimen.
  • treatment refers to any administration of a substance (e.g. , provided compositions) that partially or completely alleviates, ameliorates, relives, inhibits, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms, signs, features, and/or causes of a particular disease, disorder, and/or condition .
  • a substance e.g. , provided compositions
  • such treatment may be administered to a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition.
  • treatment may be administered to a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition.
  • treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and/or condition.
  • treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and/or condition.
  • variant refers to an entity that shows significant structural identity with a reference entity but differs structurally from the reference entity in the presence or level of one or more chemical moieties as compared with the reference entity. In many embodiments, a variant also differs functionally from its reference entity. In general, whether a particular entity is properly considered to be a "variant" of a reference entity is based on its degree of structural identity with the reference entity. As will be appreciated by those skilled in the art, any biological or chemical reference entity has certain characteristic structural elements. A variant, by definition, is a distinct chemical entity that shares one or more such characteristic structural elements.
  • a small molecule may have a characteristic core structural element (e.g., a macrocycle core) and/or one or more characteristic pendent moieties so that a variant of the small molecule is one that shares the core structural element and the characteristic pendent moieties but differs in other pendent moieties and/or in types of bonds present (single vs double, E vs Z, etc.) within the core.
  • a characteristic core structural element e.g., a macrocycle core
  • characteristic pendent moieties e.g., one or more characteristic pendent moieties
  • Macroautophagy (referred to as autophagy) is an evolutionarily conserved, regulated catabolic process that degrades cellular proteins and organelles, allowing the recycling of their biochemical components for use in energy production and biosynthetic reactions (1-3).
  • Autophagy has a role in a number of critical cell functions, including stress response, cellular quality control, tissue homeostasis, and energy production. Autophagy proceeds at a low basal level in all cells, where it is used to remove damaged proteins and organelles, particularly mitochondria, whose intracellular accumulation would be toxic.
  • autophagy has both pro- and anti- survival effects, and its role in cancer is also contextual (3, 5, 6, 17).
  • autophagy can suppress the initiation and development of early tumors, and the loss or inhibition of autophagy promotes aneuploidy and the development of the transformed phenotype (18-21).
  • inhibition of autophagy causes tumor regressions, suggesting that the autophagic process provides a survival advantage to tumors, and acts as a mechanism for overcoming stress during oncogenesis (17, 21-23).
  • pancreatic ductal adenocarcinoma cells are dependent on autophagy, genetic or pharmacologic inhibition of autophagy leads to significant growth suppression in vitro and to a robust tumor regression and prolonged survival in pancreatic cancer xenografts and genetic mouse models.
  • autophagy inhibitors have single agent antiproliferative effects in vitro and in mice on a variety of neoplastic cells, including melanoma, glioma, lymphoma, colon, and breast cancer cells (11, 12, 22-26).
  • CQ and HCQ are effective inhibitors of autophagy in vitro
  • the present invention encompasses the recognition that their in vivo efficacy may require concentrations at the upper range of tolerability (8, 9).
  • concentrations at the upper range of tolerability 8, 9.
  • Cmax peak whole blood concentrations
  • IC5 0 S for growth inhibition by HCQ ranged from 16 to 42 ⁇ in a series of tumor cell lines (11-12).
  • the low potency of CQ and HCQ may limit their efficacy in vivo, and it is uncertain if levels of HCQ that cause sufficient and sustained inhibition of autophagy will be achieved in the large number of current clinical trials.
  • the present invention provides, among other things, certain substituted chloroquinoline triazoles.
  • provided compounds inhibit cancer cell growth and induce apoptosis, and are more potent than CQ and HCQ.
  • a series of compounds that retained the 4- aminoquinoline subunit has been synthesized. Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) was used to incorporate substituted triazoles into the parent structure to form a library of novel autophagy inhibitors with increased potency compared to CQ and HCQ.
  • CuAAC Copper(I)-catalyzed azide-alkyne cycloaddition
  • Alkyne 1 was readily prepared from known alcohol 2 (scheme 2) (28). Chlorination was achieved by treatment with thionyl chloride and th pargylamine.
  • Triazoles 3a-3v were prepared by CuAAC reaction with the azides corresponding to 3a -3v (FIG. 8, SCHEME 2).
  • the symmetrical analog 6 was prepared from dipropargylamine and azide corresponding to 3r (FIG. 9, SCHEME 3a).
  • Truncated analogs 6a and 6b were prepared in analogy with compounds 3 (FIG. 9, SCHEME 3b).
  • EAD1 Growth inhibition by EAD1 (3h) was also demonstrated when the cells were treated with the drugs for 24 hours, and then allowed to form colonies in drug-free medium for 10 days. As shown in FIG. 3, EAD1 was greater than 10-fold more potent than HCQ in this assay. EAD1 induced apoptosis in the H460 cells in a concentration-dependent manner, as assessed by the detection of cell surface phosphatidylserine, a marker of early apoptosis, by fluorescently-labeled annexin V (FIG. 4). Apoptosis induced by EAD1 was significantly greater than that induced by CQ and HCQ at drug concentrations of 25, 50 and 75 ⁇ .
  • Chloroquine is a diprotic weak base and in cells it is concentrated in the acidic environment of lysosomes, as predicted by its pKs (8.1 and 10.1), where it elevates lysosomal pH through its actions as a tertiary amine (29-30). Examination of their chemical structures suggests it is unlikely that differential effects on lysosomal pH can account for the potencies of the compounds as most potent compound (3h) and the weakest inhibitor (3m) have very similar basicity (chlorobenzyl vs. propanol sidechains), yet their inhibitory activities were remarkably different.
  • Autophagy is a dynamic multistep process which is initiated when a portion of the cytoplasm containing the intended cargo is sequestered in a double membrane vesicle that eventually closes to form an autophagosome (2-5).
  • the outer membrane of the autophagosome then fuses with the lysosome, and the inner material is degraded by lysosomal enzymes which are then recycled into the cell.
  • Several proteins, known as atg proteins are required for the various steps in the formation of the autophagosome.
  • the mammalian homolog of atg8 is also known as the microtubule-associated light chain 3 (LC3).
  • LC3 is a widely used marker of autophagy, as it is either in a free cytoplasmic form (LC3-I) or as a lipidated form inserted into the inner and outer membranes of the autophagosome (LC3-II).
  • HCQ and CQ inhibit the autophagic process at a late stage, leading to the accumulation of autophagosomes and an increase in LC3-II in treated cells (31).
  • LC3 levels were also evaluated by immunoblot of drug-treated H460 cells, which allows for the distinction between cytoplasmic LC3-I and autophagosome-bound LC3-II (31). While both HCQ and EADl caused concentration and time-dependent increases in LC3-II levels (FIGS. 6a and 6b), the increase was larger at each concentration for EADl than for HCQ. The effect of 25 ⁇ HCQ on LC3-II was approximately equal to that of 5 ⁇ EADl (FIG. 6a).
  • Compounds according to the present invention may be readily formulated into pharmaceutical compositions, with a variety of uses including, in some embodiments, in the inhibition of autophagy in a biological system and/or the inhibition, treatment or prevention of diseases states and/or conditions which benefit from the inhibition of autophagy including cancer (and its metastasis), rheumatoid arthritis, malaria, antiphospholipid antibody syndrome, lupus (systemic lupus erythematosus), chronic urticaria and Sjogren's disease.
  • Pharmaceutical compositions comprise an effective amount of one or more compounds according to the present invention in combination with a pharmaceutically acceptable carrier, additive or excipient, optionally in combination with at least one additional agent, in the case of cancer, preferably an anticancer agent as otherwise described herein.
  • the compounds and method of the invention may be used to inhibit autophagy as otherwise described herein, and are useful for the inhibition (including prophylaxis) and/or treatment of cancer and its metastasis, rheumatoid arthritis, malaria, antiphospholipid antibody syndrome, lupus (systemic lupus erythematosus), chronic urticaria and Sjogren's disease.
  • the treatment of cancer or malaria are important aspects of the present invention.
  • subjects or patients in need are treated with the present compounds, pharmaceutical compositions in order to inhibit, reduce the likelihood or treat a disease state, condition and/or infection as otherwise described herein.
  • the disease states, conditions and infections treated by the present compounds and compositions are readily recognized and diagnosed by those of ordinary skill in the art and treated by administering to the patient an effective amount of one or more compounds according to the present invention.
  • dosages and routes of administration of the compound are determined according to the size and condition of the subject, according to standard pharmaceutical practices. Dose levels employed can vary widely, and can readily be determined by those of skill in the art. Typically, amounts in the milligram up to gram quantities are employed.
  • composition may be administered to a subject by various routes, e.g. orally, transdermally, perineurally or parenterally, that is, by intravenous, subcutaneous, intraperitoneal, or intramuscular injection, among others, including buccal, rectal and transdermal administration.
  • routes e.g. orally, transdermally, perineurally or parenterally, that is, by intravenous, subcutaneous, intraperitoneal, or intramuscular injection, among others, including buccal, rectal and transdermal administration.
  • Subjects contemplated for treatment according to the method of the invention include humans, companion animals, laboratory animals, and the like.
  • Formulations containing the compounds according to the present invention may take the form of solid, semi-solid, lyophilized powder, or liquid dosage forms, such as, for example, tablets, capsules, powders, sustained-release formulations, solutions, suspensions, emulsions, suppositories, creams, ointments, lotions, aerosols, patches or the like, preferably in unit dosage forms suitable for simple administration of precise dosages.
  • compositions according to the present invention typically include a conventional pharmaceutical carrier or excipient and may additionally include other medicinal agents, carriers, adjuvants, additives and the like.
  • the composition is about 0.1% to about 85%, about 0.5% to about 75% by weight of a compound or compounds of the invention, with the remainder consisting essentially of suitable pharmaceutical excipients.
  • excipients include pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, glucose, gelatin, sucrose, magnesium carbonate, and the like.
  • the composition may also contain minor amounts of non-toxic auxiliary substances such as wetting agents, emulsifying agents, or buffers.
  • Liquid compositions can be prepared by dissolving or dispersing the compounds (about 0.5% to about 20% by weight or more), and optional pharmaceutical adjuvants, in a carrier, such as, for example, aqueous saline, aqueous dextrose, glycerol, or ethanol, to form a solution or suspension.
  • a carrier such as, for example, aqueous saline, aqueous dextrose, glycerol, or ethanol
  • the composition may be prepared as a solution, suspension, emulsion, or syrup, being supplied either in liquid form or a dried form suitable for hydration in water or normal saline.
  • the preparations may be tablets, granules, powders, capsules or the like.
  • the composition is typically formulated with additives, e.g. an excipient such as a saccharide or cellulose preparation, a binder such as starch paste or methyl cellulose, a filler, a disintegrator, and other additives typically used in the manufacture of medical preparations.
  • additives e.g. an excipient such as a saccharide or cellulose preparation, a binder such as starch paste or methyl cellulose, a filler, a disintegrator, and other additives typically used in the manufacture of medical preparations.
  • An injectable composition for parenteral administration will typically contain the compound in a suitable i.v. solution, such as sterile physiological salt solution.
  • a suitable i.v. solution such as sterile physiological salt solution.
  • the composition may also be formulated as a suspension in a lipid or phospholipid, in a liposomal suspension, or in an aqueous emulsion.
  • compositions for administration as described herein contain a quantity of the selected compound in a pharmaceutically effective amount for inhibiting autophagy in a biological system, including a patient or subject according to the present invention.
  • compounds described herein are administered to a subject in need thereof, e.g., to inhibit autophagy and/or to inhibit, treat, and/or prevent one or more disease states and/or conditions including cancer (including metastasis of cancer), rheumatoid arthritis, malaria, antiphospholipid antibody syndrome, lupus, chronic urticaria and Sjogren's disease.
  • cancer including metastasis of cancer
  • rheumatoid arthritis including malaria, antiphospholipid antibody syndrome, lupus, chronic urticaria and Sjogren's disease.
  • compounds according to the present invention may be used to inhibit, reduce the likelihood or treat cancer, including the metastasis of cancer in a patient or subject in need of such treatment.
  • the cancer is one for which inhibition of autophagy represents a favorable result and/or for which metastasis is a risk element.
  • the cancer is drug resistant cancer.
  • patients or subjects in need thereof are treated by administering to the patient or subject an effective amount of one or more compounds according to the present invention, optionally in combination with at least one additional bioactive agent, e.g., that is useful (and/or approved) for treating the same disease state or condition.
  • an additional bioactive agent e.g., that is useful (and/or approved) for treating the same disease state or condition.
  • an effective amount of a provided compound may be defined as an amount equivalent to (i.e., that shows comparable activity in a relevant assay and/or under a relevant set of conditions) to a known amount of CQ or HCQ.
  • CQ and HCQ are typically dosed in amounts up to 1200 mg per day (e.g., for autophagy inhibition and/or for treatment of malaria), up to 600 mg per day (e.g., for rheumatoid arthritis), and up to 400 mg per day (e.g., for systemic lupus erythematosus). Lower doses are often employed for chronic treatment.
  • provided compounds are administered in combination with one or more additional therapeutic agents, e.g., that is/are effective and/or approved for treatment of the same disease, disorder or condition (e.g., cancer).
  • additional therapeutic agents e.g., that is/are effective and/or approved for treatment of the same disease, disorder or condition (e.g., cancer).
  • provided compounds are administered in combination with one or more anticancer agents.
  • Representative such anticancer agents may include, for example, one or more of everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HDAC inhibitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3
  • the method of treatment comprises administering to the subject in need of treatment, in a pharmaceutically acceptable carrier, an effective amount of a compound.
  • the present invention provides, among other things, various technologies for identifying and/or characterizing compounds of interest.
  • the present disclosure describes a variety of assays and systems for assessing one or more characteristics, activities, and/or attributes of compounds; those of ordinary skill in the art will appreciate that these or other available systems may be utilized, for example to identify compounds of interest by analysis of one or more test compounds and/or to characterize compounds (and/or compositions or preparations that contain them).
  • Chloroquine and hydroxychloroquine sulfate were obtained from Spectrum Chemicals and Sigma, respectively. Chloroquine analogs were dissolved in H 2 0 and were stored at -20° C.
  • Anhydrous solvents (tetrahydrofurane and toluene,) were obtained using a Pure Solv AL-258 solvent purification system. Dimethyl formamide was dried over activated 4 A molecular sieves.
  • NMR spectra were recorded on Bruker DRX 300 and DRX 600 spectrometers.
  • X H and 1 C chemical shifts ( ⁇ ) are reported relative to tetramethyl silane (TMS, 0.00/0.00 ppm) as internal standard or to residual solvent (CD 3 OD: 3.31 ppm/49.00 ppm; CDCI 3 : 7.26/77.16 ppm; DMSO-d6: 2.50/39.52 ppm).
  • ESI-MS High resolution electrospray ionization mass spectra
  • IR spectra were recorded on an Agilent Cary 630 FTIR on a ZnSe crystal. Wave numbers are reported in cm -1 .
  • Human H460, HCC827, and BxPC3 cell lines were from ATCC (Manassas, VA), and were maintained in RPMI-1640 medium with 10% fetal bovine serum, penicillin and streptomycin, at 37° C in a humidified atmosphere with 5% CO2.
  • Cells were seeded in 96 well plates in triplicate (2-4 x lOVwell) and allowed to attach for 24 hours before different concentrations of the drugs were added for an additional 48 or 72 hours.
  • Cell proliferation was quantified by a sulforhodamine B (SRB) assay. Attached cells were fixed with 10% trichloroacetic acid and incubated for 1 hour at 5° C. The cells were stained with SRB (0.4% in 1% acetic acid) by incubating at room temperature for 30 minutes. The plate was rinsed 4x with 1% acetic acid and dried. The SRB was dissolved by adding 10 mM Tris-base. Absorbances were read at 510 nm on plate reader. Cell numbers were expressed relative to untreated controls, and IC5 0 S were calculated from concentration-response graphs. Data are means ⁇ SD of at least 3 experiments.
  • H460 cells were plated at 250 cells per well in 24 well plates. After allowing for cell attachment ovemight, HCQ and EADl were added at the indicated concentrations. After 24 hours, drug-containing medium was removed, and drug- free medium added. Cell colonies were stained with crystal violet after an additional 10-day growth in the absence of drug. Apoptosis assay
  • H460 cells were cultured in medium containing 5, 25, 50 or 75 ⁇ of HCQ, CQ or EAD1 for 24 hours. For quantitative determination, cells were trypsinized, stained with Annexin V for 15 minutes at room temperature, and analyzed by flow cytometry. Each experiment was performed in triplicate.
  • Cells were scraped from culture dishes, cell lysates were prepared, and immunoblot analysis was performed.
  • Cell extracts were prepared in cold lysis buffer (50 mM Tris pH 7.5, 100 mM NaCl, 50 mM NaF, 5 mM Ethylenediaminetetraacetic acid(EDTA), 1% Triton XI 00, 200 ⁇ Na orthovanadate and protease inhibitor cocktail (HALT; Thermo Scientific)). Protein concentrations were determined using the Lowry reagent (Bio-Rad), and normalized cell lysates were mixed with sample buffer (Bio-Rad) containing 2- mercaptoethanol and boiled for 5 mins.
  • the samples were run on SDS-polyacrylamide gels and transferred to nitrocellulose or PVDF membranes.
  • the membranes were incubated overnight with primary antibodies in TBS-T buffer containing 5% non-fat milk: LC3 (Cell Signaling LC3 A/B (D3U4C) and p62 (Sigma Anti-p62/SQSTMl rabbit #P0067). After washing, the membranes were incubated with HRP-conjugated secondary antibody for 1 hour. The bands were detected with enhanced chemiluminescence reagent (Pierce).
  • the present invention relates to the compound according to intermediate: Alkyne 1, an intermediate in the synthesis of the compounds shown in chemical structures (EAD1) and others.
  • An efficient synthesis of Intermediate: Alkyne 1 based on a published procedure (16) has been established.
  • Autophagy inhibitor Lys05 has single-agent antitumor activity and reproduces the phenotype of a genetic autophagy deficiency. Proc Natl Acad Sci USA. 2012, 109,8253-8258.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des composés triazole de chloroquinoline, ainsi que des compositions comprenant les composés et des procédés associés, notamment des procédés d'inhibition de l'autophagie dans les systèmes biologiques et de traitement de maladies, de troubles ou d'affections dans lesquels l'inhibition de l'autophagie peut conférer un avantage, notamment le traitement du cancer, de la polyarthrite rhumatoïde, de la malaria, du syndrome des anticorps antiphospholipides, du lupus, de l'urticaire chronique et de la maladie de Sjogren.
PCT/US2016/012106 2015-01-06 2016-01-05 Composés triazole de chloroquinoline, composition et utilisations WO2016111957A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201562100316P 2015-01-06 2015-01-06
US62/100,316 2015-01-06

Publications (1)

Publication Number Publication Date
WO2016111957A1 true WO2016111957A1 (fr) 2016-07-14

Family

ID=56356341

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/012106 WO2016111957A1 (fr) 2015-01-06 2016-01-05 Composés triazole de chloroquinoline, composition et utilisations

Country Status (1)

Country Link
WO (1) WO2016111957A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020046335A1 (fr) * 2018-08-30 2020-03-05 Albert Einstein College Of Medicine, Inc. Composés utiles en tant que modulateurs de l'autophagie à médiation par des chaperones

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013134047A2 (fr) * 2012-03-07 2013-09-12 The Mclean Hospital Corporation Dérivés d'aminoquinoléine et leurs utilisations
US20140050696A1 (en) * 2011-04-29 2014-02-20 Ravi K. Amaravadi Novel bisaminoquinoline compounds, pharmaceutical compositions prepared therefrom and their use

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140050696A1 (en) * 2011-04-29 2014-02-20 Ravi K. Amaravadi Novel bisaminoquinoline compounds, pharmaceutical compositions prepared therefrom and their use
WO2013134047A2 (fr) * 2012-03-07 2013-09-12 The Mclean Hospital Corporation Dérivés d'aminoquinoléine et leurs utilisations

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MANOHAR ET AL.: "Synthesis of 4-aminoquinoline-1,2,3-triazole and 4-aminoquinoline- -1,2,3- triazole-1,3,5-triazine Hybrids as Potential Antimalarial Agents", CHEMICAL BIOLOGY & DRUG DESIGN, vol. 78, no. 1, 2011, pages 124 - 136 *
TALELI: "Synthesis of Triazole-Linked Chloroquinoline Derivatives as Novel Antimalarial Agents", THESIS SUBMITTED TO THE FACULTY OF NATURAL SCIENCE , UNIVERSITY OF STELLENBOSCH, March 2013 (2013-03-01), pages 1 - 240 *
TUKULULA ET AL.: "Synthesis and Antiplasmodial and Antimycobacterial Evaluation of New Nitroimidazole and Nitroimidazooxazine Derivatives", ACS MED. CHEM. LETT., vol. 4, 2013, pages 128 - 131 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020046335A1 (fr) * 2018-08-30 2020-03-05 Albert Einstein College Of Medicine, Inc. Composés utiles en tant que modulateurs de l'autophagie à médiation par des chaperones
JP2022511269A (ja) * 2018-08-30 2022-01-31 アルベルト・アインシュタイン・カレッジ・オブ・メディシン シャペロン介在性オートファジー調節剤として有用な化合物
JP7262141B2 (ja) 2018-08-30 2023-04-21 アルベルト・アインシュタイン・カレッジ・オブ・メディシン シャペロン介在性オートファジー調節剤として有用な化合物
US11834424B2 (en) 2018-08-30 2023-12-05 Albert Einstein College Of Medicine Compounds useful as chaperone-mediated autophagy modulators

Similar Documents

Publication Publication Date Title
US11098040B2 (en) Compounds and methods of use
US11639335B2 (en) Asymmetric bisaminoquinolines and bisaminoquinolines with varied linkers as autophagy inhibitors for cancer and other therapy
US20210369731A1 (en) Covalent targeting of e3 ligases
CA2993605C (fr) Derives de 1,3-benzodioxole destines au traitement ou a la prevention deleucemie a lymphocytes t ou de lymphome t chez l'adulte
US10774047B2 (en) Dimeric quinacrine derivatives as autophagy inhibitors for cancer therapy
RU2694624C2 (ru) Производные пирролидинона в качестве ингибиторов метар-2
JP2017531681A (ja) Ｎｉｋ阻害剤としての新規化合物
US11084829B2 (en) Ubiquitin-specific-processing protease 7 (USP7) modulators and uses thereof
US20220144845A1 (en) SUBSTITUTED PYRROLO[1,2-a]QUINOXALIN-4(5H)-ONES AS CX3CR1 ANTAGONISTS
CN114008024A (zh) 具有铁死亡诱导活性的化合物以及使用其的方法
CN115443276A (zh) 吡啶并嘧啶酮衍生物及其作为芳烃受体调节剂的应用
WO2012135779A1 (fr) Combinaisons de composés inhibiteurs d'akt et mek, et procédés d'utilisation
WO2020263893A1 (fr) Molécules conjuguées de cannabinoïdes
US20130338372A1 (en) Substituted Imidazoline Compounds
CN109715166A (zh) 癌症的治疗
WO2017025493A1 (fr) Inhibiteurs quinoléine d'ezh2
US10485769B2 (en) Cholesteryl ester transfer protein (CETP) inhibition in the treatment of cancer
WO2016111957A1 (fr) Composés triazole de chloroquinoline, composition et utilisations
US20230346815A1 (en) Lmp7-selective inhibitors for the treatment of blood disorders and solid tumors
US20240245677A1 (en) Selected compounds for targeted degradation of brd9
NZ725361A (en) Combinations of cancer therapeutics
CA3224123A1 (fr) Inhibition de petites molecules de l'enzyme de deubiquitination josephin domain containing 1 (josd1) en tant que therapie ciblee pour des leucemies avec la janus kinase 2 (jak2) mutant
WO2020164997A1 (fr) Combinaison d'inhibiteurs de la pi3k

Legal Events

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

Ref document number: 16735274

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16735274

Country of ref document: EP

Kind code of ref document: A1