WO2023074780A1 - Composition d'agent thérapeutique - Google Patents

Composition d'agent thérapeutique Download PDF

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WO2023074780A1
WO2023074780A1 PCT/JP2022/040088 JP2022040088W WO2023074780A1 WO 2023074780 A1 WO2023074780 A1 WO 2023074780A1 JP 2022040088 W JP2022040088 W JP 2022040088W WO 2023074780 A1 WO2023074780 A1 WO 2023074780A1
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target protein
lenalidomide
general formula
degradation
inducing
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PCT/JP2022/040088
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English (en)
Japanese (ja)
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達也 澤崎
聡士 山中
哲男 柴田
了 森下
哲 尾澤
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国立大学法人愛媛大学
株式会社テクノネットワーク四国
株式会社セルフリーサイエンス
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Priority to JP2023556623A priority Critical patent/JPWO2023074780A1/ja
Publication of WO2023074780A1 publication Critical patent/WO2023074780A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • 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
    • A61K9/02—Suppositories; Bougies; Bases therefor; Ovules
    • 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
    • A61K9/12—Aerosols; Foams
    • 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
    • A61K9/20—Pills, tablets, discs, rods
    • 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
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • 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
    • A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D495/14—Ortho-condensed systems

Definitions

  • the present invention relates to a therapeutic drug composition, particularly a novel target protein degradation-inducing compound, a composition for cancer treatment comprising the compound, and more particularly a composition for cancer treatment that avoids teratogenicity.
  • Thalidomide is a drug that was used worldwide as a sleep inducer in pregnant women over half a century ago. However, since it showed severe teratogenicity in the extremities of fetuses born from pregnant women who took it, it caused a world-wide drug-related problem called the thalidomide drug disaster. However, thalidomide has been shown to be effective against leprosy and multiple myeloma, and thalidomide and thalidomide derivatives (lenalidomide and pomalidomide) are currently approved as therapeutic agents for blood cancers such as multiple myeloma under strict safety control. It is an extremely effective drug that is used on a scale of about 1 trillion yen annually.
  • Thalidomide and thalidomide derivatives induce the degradation of various proteins by binding to cereblon (CRBN), which is one of the components of E3 ubiquitin ligase, a proteolytic enzyme. It has been shown to exhibit significant pharmacological effects and side effects.
  • SALL4 Sal-like protein 4
  • thalidomide 5-hydroxylation a protein that plays an important role in fetal limb development
  • PROTAC Protein degradation-inducing compound
  • PROTAC compounds bind E3 ubiquitin ligases and target proteins to form ternary complexes. This formation results in the target being polyubiquitinated and subject to degradation by the proteasome.
  • the target proteolysis-inducing compound detaches from the complex after the polyubiquitination reaction, and acts catalytically by repeatedly binding new targets to the E3 ligase. Due to this catalytic action, the target protein is depleted in cells by proteasome digestion after ubiquitination, resulting in cell death, and is effective in various diseases (eg, cancer).
  • Patent Document 1 discloses “a pharmaceutical composition comprising 6-hydroxylenalidomide, a salt thereof, or a solvate thereof”.
  • Patent Document 2 discloses “imido-based proteolytic modulators”. However, these documents do not disclose or suggest the target protein degradation-inducing compounds of the present invention.
  • the present inventors discovered PLZF (promyelocytic leukemia zinc finger) as a protein involved in thalidomide teratogenicity, and investigated the molecular mechanism of thalidomide teratogenicity. It was revealed that it decomposes Furthermore, the present inventors next conducted PLZF gene knockdown experiments in thalidomide-sensitive chicken embryos, and confirmed that PLZF plays an important role in limb development in chicken embryos. Furthermore, we confirmed that administration of thalidomide or 5-hydroxythalidomide to chick embryos decreased PLZF but not SALL4 in the limb buds of chick embryos.
  • PLZF promyelocytic leukemia zinc finger
  • an object of the present invention is to provide a novel target protein degradation-inducing compound, a composition for cancer treatment containing the compound, and particularly a composition for cancer treatment that does not cause serious teratogenicity.
  • the present inventors discovered that both SALL4 and PLZF The present invention was completed by discovering a thalidomide derivative that reduced the protein resolution of , and further confirming that a target protein degradation-inducing compound containing the derivative had an anticancer effect. That is, the present invention is as follows.
  • a target protein degradation-inducing compound having the following composition "Cereblon binder” - "linker” - "target protein binder”,
  • the cereblon binder is a target protein degradation-inducing compound selected from any one of compounds represented by the following general formula (1), salts thereof, and solvates thereof.
  • R1 is hydrogen, F, Br, Cl, CH3 , OCH3 , I, CF3 or OCF3
  • R L1 and R L2 are hydrogen, functional groups or atoms, but any 1 is a functional group or atom.
  • the target protein degradation-inducing compound according to the preceding item 1 which has the following configuration:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (1), salts thereof, and solvates thereof, (wherein R1 is hydrogen, F, Br, Cl, CH3 , OCH3 , I, CF3 or OCF3 , and R L1 and R L2 are hydrogen, functional groups or atoms, but any 1 is a functional group or atom.) and the target protein binder is JQ-1 or Birabresib. Targeted proteolysis-inducing compounds. 3. 3.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (7), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 4.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (8), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 5.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (6), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 6.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (3), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 7. 3.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (5), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 8. 3.
  • the target protein degradation-inducing compound according to 1 or 2 above which has the following structure:
  • the cereblon binder is selected from any one of compounds represented by the following general formula (4), salts thereof, and solvates thereof, (Where R L1 and R L2 are hydrogen, functional groups or atoms, but any one is a functional group or atom.) and the target protein degradation-inducing compound, wherein the target protein binder is Birapresib or JQ-1. 9.
  • 10. 9 The target protein degradation-inducing compound according to any one of 3 to 8 above, which has reduced teratogenicity. 11.
  • a composition for treating cancer comprising the target protein degradation-inducing compound according to any one of 1 to 10 above. 12.
  • a composition for treating cancer comprising a compound represented by the following general formula (1), a salt thereof, or a solvate thereof. (wherein R1 is hydrogen, F, Br, Cl, CH3 , OCH3 , I, CF3 or OCF3 , and R L1 and R L2 are each independently H, a functional group or an atom is.) 13.
  • composition for cancer treatment according to 12 above wherein the compound represented by the following general formula (1) is a compound represented by the general formula (8), a salt thereof, or a solvate thereof. (Where R L1 and R L2 are each independently H, a functional group, or an atom.) 15. 13.
  • composition for cancer treatment according to the preceding item 12 wherein the compound represented by the following general formula (1) is a compound represented by the general formula (3), a salt thereof, or a solvate thereof.
  • R L1 and R L2 are each independently H, a functional group, or an atom.
  • the composition for cancer treatment according to item 12, wherein the compound represented by the following general formula (1) is a compound represented by the general formula (5), a salt thereof, or a solvate thereof.
  • R L1 and R L2 are each independently H, a functional group, or an atom.
  • a therapeutic agent for chromosome 5-deficient myelodysplastic syndrome comprising a compound represented by the following general formula (5), a salt thereof, or a solvate thereof.
  • a therapeutic agent for chromosome 5-deficient myelodysplastic syndrome comprising a compound represented by the following general formula (3), a salt thereof, or a solvate thereof. (Where R L1 and R L2 are each independently H, a functional group, or an atom.)
  • the present invention includes the following. 1.
  • the cereblon binder is a target protein degradation-inducing compound selected from any one of compounds represented by the following general formula (10), salts thereof, and solvates thereof.
  • the present invention can provide a novel target protein degradation-inducing compound, a composition for cancer treatment containing the compound, and a composition for cancer treatment with reduced teratogenicity.
  • Linker example Linker example. Structure of each thalidomide derivative. Results of analysis of the affinity (degradation-inducing ability) of each thalidomide derivative for IKZF1, SALL4, and PLZF (the left column shows IKZF1, the middle column shows SALL4, and the right column shows PLZF). Results of degradation analysis of the affinity (degradation-inducing ability) of each thalidomide derivative for IKZF1, SALL4, and CRBN.
  • Results of anticancer activity against cultured cells derived from multiple myeloma of lenalidomide derivatives with substituents inserted at each meta position results of anticancer activity of lenalidomide derivatives with substitution groups inserted at each meta position against cultured cells derived from chromosome 5-deficient myelodysplastic syndrome.
  • PROTACs Examples of known PROTACs. Examples of known PROTACs. Examples of known PROTACs. Examples of known PROTACs. Examples of known PROTACs. Examples of known PROTACs. Evaluation of the antitumor effect of PROTAC in a mouse xenograft model using IMR32 cells (neuroblastoma cells). Results of the ability to induce degradation of thalidomide, pomalidomide and lenalidomide. Results of degradation induction ability in 4-position modification of thalidomide derivatives. The result of degradation induction ability by substitution of 5th position, 6th position, etc. of a pomalidomide frame
  • PROTAC 7-position modified pomalidomide-linker-JQ-1 derivative
  • composition for cancer treatment containing a compound represented by the following general formula (1) or general formula (10), a salt thereof, or a solvate thereof (hereinafter sometimes referred to as the "composition for cancer treatment of the present invention” be).
  • composition for cancer treatment of the present invention a compound represented by the following general formula (1) or general formula (10), a salt thereof, or a solvate thereof (hereinafter sometimes referred to as the "composition for cancer treatment of the present invention” be).
  • Target protein degradation-inducing compounds having the following composition: "Cereblon binder” - "linker” - "target protein binder”
  • R1 is hydrogen, F, Br, Cl, CH3 , OCH3 , I, CF3 or OCF3
  • R L1 and R L2 are hydrogen, functional groups or atoms, any one of which is a functional group or an atom.
  • R10 is hydrogen, a functional group, an atom or NR L1
  • R L2 and R L1 and R L2 are hydrogen, a functional group or an atom, any one of which is a functional group or
  • R11 is hydrogen, F, Br, Cl, CH3 , OCH3 , I, CF3 or OCF3
  • R12 is hydrogen, F, Br, CF3 , I, Cl, OH, CH 3 , OCF3 or OCH3 .
  • either one or both of RL1 and RL2 are bound to or part of the linker.
  • functional groups include alcohols, ketones, carboxylic acids, amines, ethers, etc., as long as they are capable of bonding (especially covalent bonding) or coupling with the linkers described below.
  • atoms include, but are not limited to, oxygen, nitrogen, sulfur, phosphorus, halogen, heterocyclic amines (piperazine, piperidine), etc., as long as they can bond or couple with the linker described below.
  • composition for cancer treatment of the present invention contains the compound represented by the general formula (1) or general formula (10), a salt thereof, or a solvate thereof as an active ingredient.
  • the compound represented by the general formula (1) or general formula (10) can be exemplified below.
  • the active ingredient can function as a cereblon binder.
  • Salts are not particularly limited as long as they are pharmacologically acceptable, and specifically, acid addition salts, base addition salts and the like are preferable.
  • Solvates are not particularly limited as long as they are pharmacologically acceptable, and specifically, hydrates, ethanolates and the like are preferable.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • F-lenalidomide is a lenalidomide derivative in which fluorine is attached to the 6-position of the aminoisoindolinone ring of lenalidomide.
  • F-lenalidomide can be synthesized, for example, by appropriately changing the raw material compounds, reaction steps, etc. so that the 6-position of the aminoisoindolinone ring has fluorine in a known method for synthesizing lenalidomide.
  • F-lenalidomide has S- and R-enantiomers. In the present invention, S-enantiomers, R-enantiomers, or mixtures thereof (racemic mixtures, etc.) are targeted.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • Lenalidomide is commercially available and can be produced by a method known per se.
  • Lenalidomide has S- and R-enantiomers.
  • S-enantiomers, R-enantiomers, or mixtures thereof are targeted.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • Cl-Lenalidomide is a lenalidomide derivative with a chlorine attached to the 6-position of the aminoisoindolinone ring of lenalidomide.
  • Cl-lenalidomide can be synthesized, for example, by appropriately changing the raw material compounds, reaction steps, etc. so that chlorine is present at the 6-position of the aminoisoindolinone ring in a known method for synthesizing lenalidomide.
  • Cl-lenalidomide has S- and R-enantiomers. In the present invention, S-enantiomers, R-enantiomers, or mixtures thereof (racemic mixtures, etc.) are targeted.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • Br-lenalidomide is a lenalidomide derivative with Br attached to the 6-position of the aminoisoindolinone ring of lenalidomide.
  • Br 2 -lenalidomide can be synthesized, for example, by appropriately changing starting compounds, reaction steps, etc. so that Br is present at the 6-position of the aminoisoindolinone ring in a known method for synthesizing lenalidomide.
  • Br-lenalidomide has S and R optical isomers. In the present invention, S-isomer, R-isomer, or a mixture thereof (racemic mixture, etc.) is targeted.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • F 3 C-lenalidomide is a lenalidomide derivative in which CF 3 is attached to the 6-position of the aminoisoindolinone ring of lenalidomide.
  • F 3 C-lenalidomide can be synthesized, for example, by appropriately changing the raw material compounds, reaction steps, etc. so as to have CF 3 at the 6-position of the aminoisoindolinone ring in a known method for synthesizing lenalidomide.
  • F 3 C-lenalidomide has S- and R-enantiomers. In the present invention, S-enantiomers, R-enantiomers, or mixtures thereof (racemic mixtures, etc.) are targeted.
  • R L1 and R L2 are each independently H, a functional group or an atom.
  • F 3 CO-lenalidomide is a lenalidomide derivative in which F 3 CO is attached to the 6-position of the aminoisoindolinone ring of lenalidomide.
  • F 3 CO-lenalidomide can be synthesized, for example, by appropriately changing the raw material compounds, reaction steps, etc. so that F 3 CO is present at the 6-position of the aminoisoindolinone ring in a known method for synthesizing lenalidomide.
  • F 3 CO-lenalidomide has S- and R-optical isomers, and the present invention targets the S-isomer, R-isomer, or a mixture thereof (racemic mixture, etc.).
  • thalidomide derivatives In addition to the above compounds, thalidomide derivatives, pomalidomide derivatives, and lenalidomide derivatives shown in FIGS. 35 to 38 are also targeted.
  • the target protein degradation-inducing compound of the present invention includes a structure of “cereblon binder”-“linker”-“target protein binder”.
  • the target protein degradation-inducing compound also includes a salt or solvate form thereof.
  • the salt is not particularly limited as long as it is pharmacologically acceptable, and specifically, acid addition salts, base addition salts and the like are preferable.
  • Solvates are not particularly limited as long as they are pharmacologically acceptable, but specifically, hydrates, ethanolates and the like are preferred.
  • the cereblon binder is the compound represented by general formula (1) or general formula (10), the compound represented by general formula (3), or the compound represented by general formula (4). Any compound selected from a compound represented by the general formula (5), a compound represented by the general formula (6), a compound represented by the general formula (7) and a compound represented by the general formula (8) or one or more compounds.
  • linker As the linker of the present invention, a linker known per se can be used as long as both ends of the linker can be linked (in particular, chemically (covalently) linked or coupled) to the cereblon binder and the target protein binder.
  • a linker can be linked to a cereblon binder and a target protein binder by a known technique. For example, reference can be made to the document "Frontiers in Chemistry, 9, 707317 (2021)" for a known linker binding method with PROTAC. Further, for example, the linker can be represented by the formula below.
  • each R M1 or R M2 is independently linked to another L group and can be further substituted with 0 to 4 R M5 groups, cycloalkyl and/or Heterocyclyl moieties can be formed.
  • the linkers described in FIGS. 1 and 2 can be exemplified.
  • the target protein degradation-inducing compound of the present invention can be applied to various diseases (particularly cancer types) because the degradation-inducing ability of SALL4, PLZF, and the like is reduced according to the following examples. If the target protein degradation-inducing compound of the present invention has the cereblon binder of the present invention, it can be used as an active ingredient of therapeutic agents for various diseases by containing the known target protein binders shown in FIGS. can do. For example, based on the structure of No. 1 ARV-825 shown in FIG.
  • the compound is effective against multiple myeloma (MM), diffuse large B-cell lymphoma (DLBCL), neuroblastoma, T-cell acute lymphoblastic It can be used as an active ingredient of a therapeutic agent for leukemia (T-ALL), acute myeloid leukemia (AML) or Burkitt's lymphoma.
  • T-ALL multiple myeloma
  • DLBCL diffuse large B-cell lymphoma
  • AML acute myeloid leukemia
  • Burkitt's lymphoma a therapeutic agent for leukemia
  • the compound can be used as an active ingredient of a therapeutic agent for CML (chronic myelogenous leukemia).
  • CML chronic myelogenous leukemia
  • the compound can be used as an active ingredient of a therapeutic agent for diffuse large B-cell lymphoma (DLBCL).
  • a therapeutic agent for diffuse large B-cell lymphoma DLBCL
  • the compound can be used as an active ingredient of a virus therapeutic agent.
  • the compound can be an active ingredient of an anti-inflammatory agent.
  • the compound can be used as an active ingredient for treating Alzheimer's disease and frontotemporal dementia.
  • a person skilled in the art can design the target protein degradation-inducing compound of the present invention containing the cereblon binder of the present invention based on the above exemplification and referring to the structures of known target protein degradation-inducing compounds shown in FIGS. can do.
  • target protein binder As the "target protein binder" of the present invention, target protein binders used in known PROTACs can be used, and the following can be exemplified.
  • JQ-1 (CAS No.: 1268524-70-4) OTX015 (CAS No.: 202590-98-5) Darolutamide (CAS No.: 1297538-32-9) Ceritinib (CAS No.: 1032900-25-6) Brigatinib (CAS No.: 1197953-54-0) Crizotinib (CAS No.: 877399-52-5) Alectinib (CAS No.: 1256580-46-7) Venetoclax (CAS No.: 1257044-40-8) Dasatinib (CAS No.: 302962-49-8) Bosutinib (CAS No.: 380843-75-4) RX-37 (CAS No.: 1627715-60-9) BI-882370 (CAS No.: 1392429-79-6) ibru
  • target diseases include, but are not limited to, the following. multiple myeloma (MM), diffuse large B-cell lymphoma (DLBCL), neuroblastoma, T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia (AML), Burkitt's lymphoma, castration-resistant prostate cancer (CRPC), non-small cell lung cancer (NSCLC), T-cell lymphoma (TCL), chronic myelogenous leukemia (CML), triple-negative breast cancer (TNBC) breast cancer)), malignant melanoma, mantle cell lymphoma (MCL), colorectal cancer (CRC), triple negative breast cancer (TNBC), ovarian cancer, multiple myeloma ( MM: Multiple Myeloma), acute T-cell lymphoma, breast cancer, ER (estrogen receptor)-positive breast cancer cells (ER+BC), liver cancer, prostate cancer, Hodgkin lymphoma: HL),
  • target protein degradation-inducing compounds of the present invention Preferred target protein degradation-inducing compounds of the present invention are exemplified below, but are not particularly limited.
  • the target protein degradation-inducing compound of the present invention can be synthesized by appropriately modifying a known method for synthesizing a target protein degradation-inducing compound.
  • a target protein degradation-inducing compound for example, "F-lenalidomide” - "linker” - “JQ-1 or Birabresib” is a known target protein degradation-inducing compound ARV-825 (CAS No.: 1818885-28-7) during the synthesis of " can be synthesized by substituting "F-lenalidomide” for "lenalidomide”.
  • Carcinomas for which the composition for cancer treatment of the present invention is indicated Carcinomas to which the following compounds contained as active ingredients are preferably applied have reduced teratogenicity according to the results of the following examples, and include not only hematologic cancers but also the following cancer types.
  • Teratogenicity is the property or action of certain substances such as drugs that interfere with the normal development of organisms and cause malformations, and is used synonymously with teratogenicity.
  • "Reduced teratogenicity" in the present invention refers to SALL4, PLZF, CK1 ⁇ , IKZF1 or SALL4, PLZF, CK1 ⁇ , IKZF1, or SALL4, PLZF, CK1 ⁇ , IKZF1 by directly or indirectly binding to cereblon of lenalidomide, which is the E3 ligase (cereblon) binder in lenalidomide or PROTAC compounds.
  • the resolution of SALL4 of F-Len is in the range of 1-70, preferably 5-60, more preferably 10-50 compared to lenalidomide.
  • the PLZF resolving power of F-Len is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolution of SALL4 of Cl-Len is in the range of 0.5-20, preferably 1.0-15, more preferably 1.5-10 compared to lenalidomide.
  • the PLZF resolving power of Cl-Len is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolution of IKZF1 of Cl-Len is in the range of 50-90, preferably 60-85, more preferably 65-80 compared to lenalidomide.
  • the resolving power of IKZF3 of Cl-Len is in the range of 30-70, preferably 35-65, more preferably 40-60 compared to lenalidomide.
  • the resolution of SALL4 of Br-Len is in the range of 0.5-40, preferably 1.0-35, more preferably 1.5-30 compared to lenalidomide.
  • the PLZF resolving power of Br-Len is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolution of IKZF1 of Br-Len is in the range of 15-50, preferably 20-45, more preferably 25-40 compared to lenalidomide.
  • the resolving power of IKZF3 of Br-Len is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolution of SALL4 of F 3 C-Len is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolving power of CK1 ⁇ of F 3 C-Len ranges from 0 to 90 compared to lenalidomide.
  • the resolving power of IKZF1 for F 3 C-Len ranges from 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolving power of IKZF3 for F 3 C-Len ranges from 0-30, preferably 0-20, more preferably 0-10 compared to lenalidomide.
  • the resolution of SALL4 of F-Len-PROTAC is in the range of 30-90, preferably 40-80, more preferably 50-70 compared to Len-PROTAC.
  • the PLZF resolution of F-Len-PROTAC is in the range of 0-60, preferably 5-50 compared to Len-PROTAC.
  • the resolution of IKZF3 of F-Len-PROTAC is in the range of 60-90 compared to Len-PROTAC.
  • the resolution of SALL4 of Cl-Len-PROTAC is in the range of 5-50, preferably 10-40, more preferably 15-30 compared to Len-PROTAC.
  • the PLZF resolution of Cl-Len-PROTAC is in the range of 0-70, preferably 0-60 compared to Len-PROTAC.
  • the resolution of IKZF1 for Cl-Len-PROTAC ranges from 70 to 95 compared to Len-PROTAC.
  • the resolution of IKZF3 of Cl-Len-PROTAC is in the range of 20-80, preferably 30-70, more preferably 40-60 compared to Len-PROTAC.
  • the resolution of SALL4 of F 3 C-Len-PROTAC is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to Len-PROTAC.
  • the PLZF resolution of F 3 C-Len-PROTAC is in the range of 0-30, preferably 0-15 compared to Len-PROTAC.
  • the resolution of CK1 ⁇ of F 3 C-Len-PROTAC is in the range of 0-30, preferably 0-20, more preferably 0-10 compared to Len-PROTAC.
  • the resolution of IKZF1 of F 3 C-Len-PROTAC is in the range of 5-50, preferably 10-40, more preferably 15-35 compared to Len-PROTAC.
  • the resolution of IKZF3 of F 3 C-Len-PROTAC is in the range of 10-70, preferably 20-60, more preferably 30-50 compared to Len-PROTAC.
  • the present inventors have confirmed that thalidomide derivatives with reduced proteolytic degradation of SALL4 or PLZF have reduced teratogenicity (Ref: EMBO J(2021) 40:e105375).
  • composition for cancer treatment of the present invention can be administered orally or parenterally.
  • known dosage forms such as tablets, capsules, coated tablets, troches, and liquids such as solutions or suspensions can be used.
  • parenteral administration includes intravenous, intramuscular, or subcutaneous administration by injection, transmucosal administration such as nasal and oral cavity administration using sprays and aerosols, rectal administration using suppositories, and patches and liniments. and transdermal administration using gel or the like.
  • Oral administration, nasal administration, or intravenous administration by injection is preferred.
  • composition for treating cancer of the present invention may contain an appropriate pharmaceutically acceptable carrier well known to those skilled in the art, depending on the mode of administration, etc., in addition to the active ingredient.
  • Pharmaceutically acceptable carriers include antioxidants, stabilizers, preservatives, flavoring agents, coloring agents, solubilizers, solubilizers, surfactants, emulsifiers, antifoaming agents, viscosity modifiers, and gelling agents. , absorption enhancers, dispersants, excipients, and pH adjusters.
  • composition for cancer treatment of the present invention is prepared as an injection preparation, it is preferably in the form of a solution or suspension preparation. Alternatively, the form of an aerosol formulation is preferred.
  • semi-solid formulations such as creams or suppositories are preferred. Any of these formulations may be prepared by any method known to those skilled in the pharmaceutical arts, for example, as described in Remington's Pharmaceutical Sciences (Mack Publishing Company, Easton, PA, 1970). can be done.
  • Injection formulations can contain, as carriers, plasma-derived proteins such as albumin, amino acids such as glycine, and sugars such as mannitol, and can also contain buffers, solubilizers, isotonic agents, and the like. can. Further, when used as an aqueous formulation or a freeze-dried formulation, it is preferable to add a surfactant such as Tween (registered trademark) 80 or Tween (registered trademark) 20 to prevent aggregation.
  • parenteral dosage forms other than injectable formulations may contain distilled water or physiological saline, polyalkylene glycols such as polyethylene glycol, oils of plant origin, hydrogenated naphthalene, and the like. good.
  • formulations for rectal administration such as suppositories contain common excipients such as polyalkylene glycols, petroleum jelly, and cocoa oil.
  • Vaginal formulations may also contain absorption enhancers such as bile salts, ethylenediamine salts and citrates.
  • Formulations for inhalation may be solid and may contain excipients such as lactose, and nasal drops may be water or oil solutions.
  • the exact dosage and dosage regimen of the composition for treating cancer of the present invention may be adjusted depending on the individual subject's requirement, treatment method, degree of disease or need, and the like. Specifically, the dosage can be determined according to age, body weight, general health condition, sex, diet, administration time, administration method, excretion rate, drug combination, patient's medical condition, and the like. may be determined in consideration of the factors of The daily dosage of the composition for treating cancer of the present invention varies depending on the condition and body weight of the patient, the type of compound, administration route and the like. 01-1000 mg/person/day, preferably 0.1-500 mg/person/day; Dosing in mg/person/day is preferred.
  • FLAG-GST-IKZF1,-SALL4,-PLZF and N-terminally biotinylated bls-CRBN were synthesized as recombinant proteins using a wheat cell-free protein synthesis system.
  • a 10 ⁇ l mixed solution containing 0.5 ⁇ l of the synthesized bls-CRBN was prepared under AlphaScreen buffer (100 mM Tris (pH 8.0), 0.01% Tween20, 100 mM NaCl, and 1 mg/mL BSA).
  • a 5 ⁇ l mixture containing 0.8 ⁇ l FLAG-GST-protein was prepared in AlphaScreen buffer.
  • a 5 ⁇ l mixture containing DMSO (final concentration 0.5%) or a thalidomide derivative at the final concentration shown in FIG. 4 was prepared in AlphaScreen buffer. The three mixtures were then mixed in a 384-well AlphaPlate (PerkinElmer) and allowed to stand at 26°C for 1 hour. Next, 5 ⁇ l detection mixture containing 0.2 ⁇ g/ml anti-DYKDDDDK antibody (FUJIFILM WakoPure Chemical), 0.08 ⁇ l streptavidin donor beads (PerkinElmer) and 0.08 ⁇ l protein A acceptor beads (PerkinElmer). was adjusted under AlphaScreen buffer. Then, 5 ⁇ l of the detection mixture was added to each well, and after standing at 26°C for 1 hour, luminescence signals were detected using an Envision plate reader (PerkinElmer).
  • FIG. 3 shows the structure of each thalidomide derivative used in this example. As is clear from the results of FIG. 4, it was confirmed that insertion of a substituent at the meta position of lenalidomide reduces the affinity (degradation-inducing ability) for SALL4 and PLZF, which are teratogenic proteins.
  • HEK293T cultured cells were seeded in a 48-well plate for cell culture (BD falcon) and cultured overnight at 37°C under 5% CO 2 conditions.
  • AGIA-SALL4 and Myc-IKZF1 were then transfected using polyethyleneimine (PEI) Max (MW 40,000) (PolyScience, Inc.).
  • PEI polyethyleneimine
  • DMSO final concentration 0.1%) or thalidomide derivatives at the final concentrations indicated in FIG. 5 were administered.
  • the medium supernatant was removed with an aspirator, and the cultured cells were collected and lysed using 1 ⁇ SDS sample buffer (62.5 mM Tris-HCl pH 6.8, 2% SDS, 10% glycerol).
  • the recovered cell extract was boiled at 95°C for 5 minutes and electrophoresed using a polyacrylamide gel. Next, immunoblotting was performed and each protein was detected using the antibodies shown in FIG.
  • FLAG-GST-IKZF1, -SALL4, -PLZF and N-terminally biotinylated bls-CRBN were synthesized as recombinant proteins using a wheat cell-free protein synthesis system.
  • a 10 ⁇ l mixed solution containing 0.5 ⁇ l of the synthesized bls-CRBN was prepared under AlphaScreen buffer (100 mM Tris (pH 8.0), 0.01% Tween20, 100 mM NaCl, and 1 mg/mL BSA).
  • AlphaScreen buffer 100 mM Tris (pH 8.0), 0.01% Tween20, 100 mM NaCl, and 1 mg/mL BSA.
  • a 5 ⁇ l mixture containing 0.8 ⁇ l FLAG-GST-protein was prepared in AlphaScreen buffer.
  • a 5 ⁇ l mixture containing DMSO (final concentration 0.5%) or a thalidomide derivative at the final concentration shown in FIG. 6 was prepared in AlphaScreen buffer. The three mixtures were then mixed in a 384-well AlphaPlate (PerkinElmer) and allowed to stand at 26°C for 1 hour. Next, 5 ⁇ l detection mixture containing 0.2 ⁇ g/ml anti-DYKDDDDK antibody (FUJIFILM WakoPure Chemical), 0.08 ⁇ l streptavidin donor beads (PerkinElmer) and 0.08 ⁇ l protein A acceptor beads (PerkinElmer). was adjusted under AlphaScreen buffer. Then, 5 ⁇ l of the detection mixture was added to each well, and after standing at 26°C for 1 hour, luminescence signals were detected using an Envision plate reader (PerkinElmer).
  • meta-substituent insertion of lenalidomide is more effective than insertion of meta-substituents of thalidomide and meta-substituent insertion of pomalidomide, which are teratogenic proteins SALL4 and It reduced the affinity for PLZF and maintained the affinity for IKZF1, a therapeutic target protein for blood cancer. Accordingly, lenalidomide and lenalidomide derivatives with substituent insertion at each meta position were used in the following examples. Although it is necessary to maintain affinity to IKZF for blood cancers, in general, affinity to IKZF is unnecessary.
  • lenalidomide derivatives with substituents inserted at each meta position were synthesized. Details are as follows. Lenalidomide was purchased from FUJIFILM WakoPure Chemical. 5-hydroxythalidomide (5HT) was synthesized by the method disclosed in Bioorg. Med Chem Lett 19:3973-3976.
  • a 10 ⁇ l mixture containing 0.5 ⁇ l of the synthesized bls-CRBN was prepared under AlphaScreen buffer (100 mM Tris (pH 8.0), 0.01% Tween20, 100 mM NaCl, and 1 mg/mL BSA).
  • a 5 ⁇ l mixture containing 0.8 ⁇ l FLAG-GST-protein was prepared in AlphaScreen buffer.
  • a 5 ⁇ l mixture containing DMSO (final concentration 0.5%) or a thalidomide derivative at the final concentration shown in FIG. 7 was prepared in AlphaScreen buffer. The three mixtures were then mixed in a 384-well AlphaPlate (PerkinElmer) and allowed to stand at 26°C for 1 hour.
  • the affinities of F3C -lenalidomide, F3CO -lenalidomide, pomalidomide and lenalidomide to IKZF1, SALL4 and PLZF were calculated. Details are as follows. FLAG-GST-IKZF1, -SALL4, -PLZF and N-terminally biotinylated bls-CRBN were synthesized as recombinant proteins using a wheat cell-free protein synthesis system.
  • a 10 ⁇ l mixed solution containing 0.5 ⁇ l of the synthesized bls-CRBN was prepared under AlphaScreen buffer (100 mM Tris (pH 8.0), 0.01% Tween20, 100 mM NaCl, and 1 mg/mL BSA).
  • a 5 ⁇ l mixture containing 0.8 ⁇ l FLAG-GST-protein was prepared in AlphaScreen buffer.
  • a 5 ⁇ l mixture containing DMSO (final concentration 0.5%) or 10 ⁇ M thalidomide derivative was prepared in AlphaScreen buffer. The three mixtures were then mixed in a 384-well AlphaPlate (PerkinElmer) and allowed to stand at 26°C for 1 hour.
  • Thalidomide-O-COOH thalidomide immobilization
  • FG-beads magnetic beads for immobilization
  • 4 mM Thalidomide-O-COOH was immobilized on FG-beads using Tamagawa Seiki's standard protocol.
  • FLAG-GST-CRBN was also synthesized using a wheat cell-free protein synthesis system.
  • IP Lysis buffer 25 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 5% adjusted to 500 ⁇ l with glycerol. After rotating at room temperature for 2 hours, it was washed four times with 800 ⁇ l of IP Lysis buffer (Pierce).
  • IP Lysis buffer (Pierce) containing 200 ⁇ M of various lenalidomide derivatives was added, and vortexing was performed for 30 minutes at room temperature for competitive elution. After that, 20 ⁇ l of 2 ⁇ SDS sample buffer was added to each eluted sample and boiled. Next, after electrophoresis using a polyacrylamide gel, immunoblotting was performed, and each protein was detected using the antibodies shown in FIG.
  • RIPA buffer 25 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA
  • Protease inhibitor cocktail Sigma Aldrich
  • the cells were cultured with fresh medium containing DMSO or lenalidomide derivatives every 3 days for a total of 9 days of culture. After that, the cells were suspended, 40 ⁇ l each was added to a 96-well opti-plate (PerkinElmer), and 40 ⁇ l each of Cell Titer-Glo kit (Promega) was added to lyse the cells. Luminescent signals were then detected using SpectraMax iD3 (Molecular devise).
  • the results in FIG. 11 are as follows.
  • the anti-proliferative activity of F-lenalidomide in cultured multiple myeloma cells is comparable to that of pomalidomide and lenalidomide, which are known active ingredients for the treatment of multiple myeloma. confirmed.
  • Cl-lenalidomide has an anti-proliferative effect, though weaker than pomalidomide and lenalidomide.
  • MDS-L cultured cells were seeded on a 24-well plate for cell culture (BD falcon), DMSO (0.1%) or the drug was administered at the lenalidomide derivative concentration shown in FIG .
  • cultured for 96 hours under Half the amount of cells was collected by pipetting, centrifuged at 200 xg for 3 minutes, and the supernatant was removed with an aspirator. After that, the cell pellet was resuspended in medium, treated with DMSO (0.1%) or lenalidomide derivative concentrations shown in FIG. 12, and cultured at 37° C., 5% CO 2 for 96 hours.
  • the cells were cultured with fresh medium containing DMSO or lenalidomide derivatives every 4 days for a total of 20 days.
  • the results in FIG. 12 are as follows.
  • the ability of F-lenalidomide to suppress the proliferation of chromosome 5-deficient myelodysplastic syndrome cultured cells is based on the anti-proliferative effect of lenalidomide, which is a known active ingredient in the treatment of chromosome 5-deficient myelodysplastic syndrome. confirmed to be strong. It was also confirmed that Cl-lenalidomide has an anti-proliferative effect comparable to that of lenalidomide. Accordingly, the subject of the present invention is a therapeutic agent for chromosome 5-deficient myelodysplastic syndrome containing F-Len or Cl-Len.
  • HuH-7 cultured cells were seeded in a 24-well plate for cell culture (BD falcon) and cultured at 37°C, 5% CO 2 for 24 hours.
  • DMSO 0.1%), 20 ⁇ M thalidomide, 10 ⁇ M lenalidomide, 1 ⁇ M pomalidomide, 20 ⁇ M 5-hydroxythalidomide, 1 ⁇ M dBET1 or 1 ⁇ M ARV-825 were administered and kept at 37°C, 5% CO 2 cultured for 24 hours under these conditions.
  • the results in FIG. 13 are as follows. Regarding dBET1, it was confirmed that the degradation-inducing ability of the teratogenic proteins SALL4 and PLZF is also low, but the degradation-inducing ability of the therapeutic targets IKZF1 and IKZF3 is also low. That is, they confirmed the problem that the anti-cancer cell proliferation effect is low. Regarding ARV-825, it was confirmed that the ability to induce the degradation of IKZF1 and IKZF3 is high, and that the ability to induce the degradation of SALL4 and PLZF is also high. In other words, we have confirmed the problem that it cannot be applied to cancer treatment in which teratogenicity is a problem. That is, it was confirmed that known PROTACs induce decomposition of not only the target protein but also the neosubstrate of the E3 binder, or cause teratogenicity.
  • Examples 1 to 10 The results of Examples 1 to 10 confirmed the following. Lenalidomide, F-lenalidomide, and Cl-lenalidomide, which degrade both IKZF1/3/CK1 ⁇ , a neo-substrate of E3 binder, and target proteins, are thought to be cereburon binders of target proteolysis-inducing compounds that are effective against blood cancer. (See FIG. 14, left), which was confirmed in the following examples.
  • Cl-lenalidomide which only degrades the target protein and has low ability to induce decomposition of the neo-substrate
  • Br-lenalidomide, F 3 C-lenalidomide, and F 3 CO-lenalidomide which do not have the ability to induce the decomposition of the neo-substrate, are neo-substrates of the E3 binder.
  • a target protein degradation-inducing compound that has low or no ability to induce degradation of a certain IKZF1/3/CK1 ⁇ , and thus can be used for other than specific cancers. cereblon binder (see right side of FIG. 14), which was confirmed in the following examples.
  • Target protein degradation-inducing compound of the present invention (Synthesis of target protein degradation-inducing compound of the present invention)
  • the target protein degradation-inducing compounds of the present invention shown in FIG. 15 were synthesized. Details are as follows. ARV-825 and dBET1 were commercially available products.
  • Lenalidomide-Linker-JQ-1 derivative (1a) was synthesized by the following procedure.
  • flow rate 5.0 mL/min
  • measurement wavelength 220 nm
  • tR 53.6 min.
  • Fluorolenalidomide-Linker-JQ-1 derivative (1b) was synthesized by the following procedure.
  • 6-fluorolenalidomide 50 mg, 0.180 mmol, 1.0 equiv
  • 2 92.0 mg, 0.216 mmol, 1.2 equiv
  • DMF 0.1 M, 2 mL
  • N,N -Diisopropylethylamine 92 ⁇ L, 0.540 mmol, 3.0 equiv
  • azeotroping with toluene was performed under reduced pressure and the solvent was distilled off.
  • flow rate 5.0 mL/min
  • measurement wavelength 220 nm.
  • 64.0 mg of yellow solid 4b was obtained with a yield of 81% (2steps).
  • MM1.S or H929 cultured cells were seeded in a 24-well plate for cell culture (BD falcon), DMSO (0.1%) or drugs were administered at concentrations shown in FIG . cultured for 24 hours under After that, the cells were collected by pipetting, centrifuged at 900 ⁇ g for 3 minutes, and the supernatant was removed with an aspirator. After that, 500 ⁇ l of 1 ⁇ PBS was added to the cell pellet, washed, centrifuged again at 900 ⁇ g for 3 minutes, and the supernatant was removed with an aspirator. Next, 100 ⁇ l of RIPA buffer containing Protease inhibitor cocktail (Sigma Aldrich) was added to the cell pellet to lyse the cells.
  • RIPA buffer containing Protease inhibitor cocktail Sigma Aldrich
  • the target protein degradation-inducing compound of the present invention does not induce the degradation of IKZF1/3/CK1 ⁇ , so it has neosubstrate selectivity for cereblon binders and has anticancer activity ( In particular, it was confirmed to have a multiple myeloma growth inhibitory effect.
  • the target protein degradation-inducing compound of the present invention can induce degradation of the target proteins BRD2, BRD3 and BRD4.
  • the target protein degradation-inducing compound of the present invention has not only multiple myeloma growth inhibitory activity but also various cancer cell growth inhibitory activities such as prostate cancer, Burkitt's lymphoma, NUT median adenocarcinoma, and lung cancer.
  • NTERA cultured cells were seeded in a 24-well plate for cell culture (BD falcon) and cultured at 37°C, 5% CO 2 for 24 hours.
  • DMSO 0.1%) or the drug at the concentration shown in FIG. 17 was administered, and cultured for 24 hours under conditions of 37° C. and 5% CO 2 .
  • the medium supernatant was removed with an aspirator, 500 ⁇ l of 1 ⁇ PBS was added to the cell pellet, and after washing, the supernatant was removed with an aspirator.
  • 100 ⁇ l of RIPA buffer containing Protease inhibitor cocktail (Sigma Aldrich) was added to the cells to lyse the cells.
  • the cells were collected by pipetting, centrifuged at 900 ⁇ g for 3 minutes, and the supernatant was removed with an aspirator. After that, 500 ⁇ l of 1 ⁇ PBS was added to the cell pellet, washed, centrifuged again at 900 ⁇ g for 3 minutes, and the supernatant was removed with an aspirator. Next, 100 ⁇ l of RIPA buffer containing Protease inhibitor cocktail (Sigma Aldrich) was added to the cell pellet to lyse the cells. Then, after centrifugation at 16,100 ⁇ g for 15 minutes, the supernatant was obtained as a cell extract.
  • RIPA buffer containing Protease inhibitor cocktail Sigma Aldrich
  • the results are shown in FIG.
  • the target protein degradation-inducing compounds of the present invention (particularly Cl-PROTAC and F 3 C-PROTAC) do not induce the degradation of SALL4 and PLZF, unlike the known PROTAC ARV-825, and do not induce the degradation of the cereblon binder neo It was confirmed to have substrate selectivity. That is, the target protein degradation-inducing compounds of the present invention (particularly Cl-PROTAC, F 3 C-PROTAC) can be used for various carcinomas by using various target protein binders.
  • HCT116 or NTERA cultured cells were seeded in a 96-well plate for cell culture (BD falcon), DMSO (0.1%) or drugs were administered at concentrations shown in FIG. Cultured for 2 days. After that, the medium supernatant was removed with an aspirator, and 40 ⁇ l of medium was added. Next, add 40 ⁇ l of Cell Titer-Glo kit (Promega) to lyse the cells, add 40 ⁇ l to 384-well opti-plate (PerkinElmer), and use SpectraMax iD3 (Molecular devise) to signal the luminescence. detected.
  • CC-220 (Degradation evaluation of SALL4 in CC-220)
  • the resolution of SALL4 of CC-220 was measured with reference to the measurement method of the above example. It was confirmed that CC-220 weakly decomposes SALL4 at concentrations capable of decomposing IKZF1 and IKZF3 (see FIG. 19). That is, CC-220 was confirmed to be an active ingredient of a composition for treating blood cancer.
  • the following fluorinated CC-220, CC-92480, and fluorinated CC-92480 which have the same chemical structure and chemical properties as CC-220, can also be the active ingredients of the composition for treating blood cancer. .
  • CC-220, fluorinated CC-220, CC-92480 and fluorinated CC-92480 can serve as cereblon binders for the targeted proteolysis-inducing compounds of the present invention.
  • the IMR32 cultured cells were seeded on a 10 cm dish for cell culture (BD falcon) and cultured under conditions of 37°C and 5% CO 2 .
  • 5 ⁇ 10 6 IMR32 cells per nude mouse were then resuspended in 50 ⁇ l PBS and mixed with 50 ⁇ l Matrigel (Corning).
  • 100 ⁇ l of the mixed cell solution was subcutaneously implanted in the flank of nude mice.
  • DMSO or PROTAC was intraperitoneally administered once a day (weekdays only).
  • the administration solution was diluted with PBS to 10% DMSO, 5 mg/kg PROTAC and 1% Tween 80, and 200 ⁇ l was administered intraperitoneally.
  • the tumor volume was measured with an electronic caliper, and the tumor was taken out and photographed.
  • FIG. 33 The results in FIG. 33 are as follows. It was confirmed that PROTAC with lenalidomide or lenalidomide derivatives suppresses tumor growth in mice. In particular, PROTAC using F 3 C-lenalidomide showed an equal or greater tumor growth inhibitory effect than PROTAC using lenalidomide.
  • Cereblon is a component of ubiquitin ligase, which is responsible for protein degradation.
  • a thalidomide derivative compound binds to cereblon, the substrate specificity of cereblon changes. A medicinal effect or side effect occurs.
  • the inventors have discovered that the induction of degradation by the binding of SALL4 and PLZF to the cereblon-thalidomide derivative compound complex, as described above, is an important event for the development of teratogenicity.
  • thalidomide-based inducers thalidomide, pomalidomide, lenalidomide
  • target degradation inducers using the thalidomide skeleton, if they have binding activity with SALL4 and PLZF, eventually degrade those proteins and cause teratogenicity.
  • Kroenke and Lu have reported on the antitumor effect of lenalidomide (cytostatic effect on multiple myeloma). It has been clarified that the anti-tumor effect of Aeolus (IKZF3) is expressed by a series of mechanisms from substrate recognition by cereblon to ubiquitination and proteasome degradation.
  • the glutarimide ring is mainly used for binding to cereblon (Reference: Nature 2014, 512, 49-53, Nature Structural & Molecular Biology 2014; We investigated the degree of degradation of IKZF1, IKZF3, SALL4 and PLZF in the medium.
  • a function target protein binder
  • BRD was selected as the degradation target protein and JQ-1 was used as the target protein binder. Furthermore, since this linker and target protein binder portion are considered to have a high sterically excluded volume effect, similarly, the ability to bind to IKZF1, IKZF3, SALL4 and PLZF and the degree of degradation thereof in cells were examined.
  • thalidomide For thalidomide, pomalidomide, and lenalidomide, the ability to bind to each molecule and the degree of degradation in cells were evaluated.
  • the binding ability to each molecule was analyzed by biochemical interaction (AlphaScreen method), and the intracellular resolution of each molecule was analyzed by the following method.
  • H929 cells The H929 cell solution was collected in an arbitrary 1.5 mL tube. Since H929 cells are floating cells, no peeling operation is required. In order to equalize the number of cells in each collected sample, it was quantified by the BCA method and diluted according to the sample with the lowest concentration. An SDS sample buffer containing mercaptoethanol was added to the diluted H929 cell solution and heated at 98° C. for 5 minutes. 2) Huh cells HuH7 cell suspension medium was aspirated. Since HuH7 cells are adherent cells, they stick to the bottom of the plate. 100 ⁇ L each of SDS sample buffer containing mercaptoethanol was placed in a 24-well plate.
  • the HuH7 cells were detached by scraping the bottom of the 24-well plate with a tip with a wide tip and collected in an arbitrary 1.5 mL tube.
  • the cells were quantified by the BCA method, the cell numbers were adjusted, and then heated at 98° C. for 5 minutes.
  • IKZF1, IKZF3, SALL4, PLZF by Western blotting IKZF1, IKZF3, SALL4, and PLZF remaining in each cell after degradation were quantified.
  • Detection photography of chemiluminescence was performed with an Amersham ImageQuant 800/Cytiva. After detection, the membrane was stripped of antibodies with a stripping solution, and the following antibody reaction was performed. Image data taken with the Amersham ImageQuant 800 was loaded and analyzed with Quantity One/Bio-Rad software.
  • Quantify the band of the target protein with Quantity One set the residual amount of each molecule after adding DMSO for evaluation control without compound as 100%, and calculate the relative ratio of the number of remaining molecules after addition of the compound as the ratio of the number of remaining molecules after degradation. (%) was used to evaluate the degree of decomposition.
  • the antibodies used for detection are shown in Table 3 below.
  • Anti-CRBN-Ab was used to confirm whether the addition of the compound increased or decreased the expression level of CRBN in the cells. It was confirmed that it was expressed in The respective antibodies are commercially available from Thermo Fisher, SANTA Cruz, Cell Signaling, MBL.
  • the results are shown in FIG.
  • the biochemical interaction of BRD the interaction between Flag-GST-BRD4 and biotinylated CRBN was evaluated by the AlphaScreen method, similar to IKZF et al.
  • the BRD antibodies in Table 3 were used, and the average value of the degradation of BRD2, BRD3, and BRD4 was obtained.
  • a 7-position modified pomalidomide-linker-JQ-1 was added to the 7-position modified pomalidomide backbone that does not induce degradation of any of IKZF1, IKZF3, SALL4, and PLZF using the same chemical reaction as in Example 12.
  • Derivatives were synthesized and biochemical interactions (AlphaScreen method) between IKZF1, IKZF3 and SALL4, PLZF, BRD4 and CRBN and the degree of degradation of each protein in cells were evaluated (see FIG. 38).
  • Both the 7-fluoropomalidomide-linker-targeting binder (JQ-1) and the 7-trifluoropomalidomide-linker-targeting binder (JQ-1) have low binding capacity to SALL4, PLZF, resulting in cell It was confirmed that the degree of degradation of SALL4 and PLZF in the medium was greatly reduced compared to the pomalidomide linker-targeted binder (JQ-1). It was confirmed that this tendency was inherited as a property of the ability to bind to IKZF1, IKZF3, SALL4 and PLZF of the basic skeleton before substitution with "linker” and "target binder” at position 4. As a result, when these compounds are used as CRBN binders, target protein degradation-inducing agents can be created that only induce degradation of the target protein and are biologically neutral in terms of other properties.
  • the therapeutic targets are both the neosubstrates IKZF1/3, CK1 ⁇ and the target proteins BRD2/3/4, so compounds with high resolution to IKZF1/3 are considered to have high therapeutic effects.
  • IKZF1/3 and CK1 ⁇ are not targeted for drug efficacy, and rather are thought to lead to side effects in the blood system. More specifically, IKZF1/3 are involved in the growth of blood cancers (especially multiple myeloma), and are proteins expressed in blood cells (especially white blood cells). presumably not significantly involved.
  • IKZF1/3 are extremely important transcription factors in the differentiation and function of hematopoietic stem cells into leukocytes (especially lymphocytes such as T cells and B cells). Indeed, IKZF1/3 knockout mice are immunodeficient and lack T and B cells.
  • IKZF1/3 knockout mice are immunodeficient and lack T and B cells.
  • degradation of IKZF1 and IKZF3 by administration of lenalidomide in mice causes a decrease in white blood cells and platelets.
  • leukocyte decrease and the like as side effects associated with the administration of thalidomide derivatives to patients.
  • significant degradation of IKZF1/3 in cancer types other than hematologic cancers does not contribute to drug efficacy and is thought to lead to side effects.
  • SALL4 is genetically shown to cause a congenital disease called Okihiro syndrome (Duane-radial ray syndrome) by mutation even in heterozygotes.
  • Okihiro syndrome Duane-radial ray syndrome
  • the disease is teratogenic at various sites, including the hands and feet, and the phenotype is very similar to thalidomide fetal disease.
  • the ability of thalidomide and thalidomide derivatives to induce degradation of SALL4 is extremely high, and Okihiro syndrome is induced even if the SALL4 mutation is heterozygous. be done.
  • administration of thalidomide to rabbits reduces SALL4 in fetuses.
  • PLZF has been reported to cause teratogenicity by mutation in humans and mice.
  • PLZF is induced to degrade in a system using chicken embryos in which SALL4 is not induced to degrade.
  • overexpression of PLZF in chicken embryos reduced thalidomide-induced teratogenicity.
  • significant teratogenicity may be caused by significant degradation of both SALL4 and PLZF in humans.
  • double knockout of SALL4 and PLZF in mice results in a more pronounced teratogenic phenotype, resembling thalidomide fetal disease.
  • F-Len has higher activity against multiple myeloma and significantly higher activity against 5qMDS than the existing drug lenalidomide.
  • the ability to induce degradation of SALL4 and PLZF is lower than that of lenalidomide.
  • F-Len is a thalidomide derivative with lower teratogenicity and higher efficacy than the existing drug lenalidomide.
  • Cl-Len has weaker activity against multiple myeloma and 5qMDS than the existing drug lenalidomide, but it does not have the ability to induce degradation of SALL4 and PLZF.
  • Cl-Len has weaker efficacy than the existing drug lenalidomide, but significantly reduces teratogenicity.
  • Br-Len has weak efficacy against multiple myeloma compared to the existing drug lenalidomide, and does not have the ability to induce degradation of SALL4 and PLZF.
  • Br-Len has weaker efficacy than the existing drug lenalidomide, but significantly reduces teratogenicity.
  • F 3 C-Len does not induce degradation of representative neo-substrates such as IKZF1/3 and CK1 ⁇ , which are therapeutic targets for hematologic cancer, and SALL4 and PLZF involved in teratogenicity.
  • F 3 C-Len is expected to be applied to PROTACs with few side effects targeting various cancer types.
  • PROTACs using F-Len have similar effects to existing PROTACs against hematological cancers, and their ability to induce degradation of SALL4 and PLZF is reduced.
  • F-Len-PROTAC is highly effective and less teratogenic against blood cancers.
  • PROTACs using Cl-Len are weaker than existing PROTACs against hematological cancers, but do not have the ability to induce the degradation of SALL4 and PLZF.
  • Cl-Len-PROTAC is effective and significantly less teratogenic against blood cancers.
  • F 3 C-LenPROTAC is effective against a variety of cancer types and appears to be free of both teratogenic and hematologic side effects.
  • F 3 CO-Len-PROTAC is effective against a variety of cancer types, both teratogenic and hematological, due to its similar chemical structure and chemical properties to F 3 C-Len-PROTAC and the results of the examples. It is considered that there are no side effects in Based on the results of Examples, Br-Len-PROTAC, like F 3 C-Len-PROTAC, is considered to be effective against various cancer types and to have no side effects in both teratogenicity and blood system.
  • a novel target protein degradation-inducing compound, a composition for cancer treatment containing the compound, and a composition for cancer treatment with reduced teratogenicity can be provided.

Abstract

Le problème décrit par la présente invention est de fournir un nouveau composé induisant la dégradation de protéine cible, une composition pour une thérapie anticancéreuse qui comprend ledit composé, et en particulier une composition pour une thérapie anticancéreuse qui ne provoque pas de tératogénicité grave. La solution selon l'invention porte sur un dérivé de thalidomide qui démontre une dégradation réduite des protéines SALL4 et PLZF qui servira de liant de ligase E3 d'un composé induisant la dégradation de protéine cible qui est capable d'éviter une tératogénicité grave. La solution proposée par l'invention a été obtenue par la découverte d'un dérivé de thalidomide qui a une capacité de dégradation réduite vis-à-vis des protéines SALL4 et PLZF, et la confirmation qu'un composé induisant la dégradation de protéine cible qui comprend ledit dérivé a un effet anticancéreux.
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WO2017197056A1 (fr) * 2016-05-10 2017-11-16 C4 Therapeutics, Inc. Dégronimères ciblant un bromodomaine pour la dégradation de protéines cibles
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JP2019526616A (ja) * 2016-09-13 2019-09-19 ザ リージェンツ オブ ザ ユニヴァシティ オブ ミシガン Ｂｅｔタンパク質分解物質としての縮合１，４−ジアゼピン
WO2020118098A1 (fr) * 2018-12-05 2020-06-11 Vividion Therapeutics, Inc. Isoindolinones substituées utilisées en tant que modulateurs du recrutement de néo-substrat à médiation par céréblon
JP2021020873A (ja) * 2019-07-26 2021-02-18 国立大学法人 奈良先端科学技術大学院大学 レナリドマイド誘導体及び医薬組成物
JP2021147365A (ja) * 2020-03-23 2021-09-27 国立大学法人 名古屋工業大学 含フッ素サリドマイド誘導体及び医薬組成物とその製造法
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JP2017513862A (ja) * 2014-04-14 2017-06-01 アルビナス インコーポレイテッド イミド系タンパク質分解モジュレーター及び関連する使用方法
WO2017197056A1 (fr) * 2016-05-10 2017-11-16 C4 Therapeutics, Inc. Dégronimères ciblant un bromodomaine pour la dégradation de protéines cibles
JP2019526616A (ja) * 2016-09-13 2019-09-19 ザ リージェンツ オブ ザ ユニヴァシティ オブ ミシガン Ｂｅｔタンパク質分解物質としての縮合１，４−ジアゼピン
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