WO2021251756A1 - N-acylhydrazone-derivative-marker conjugate and pharmaceutical composition containing same - Google Patents

N-acylhydrazone-derivative-marker conjugate and pharmaceutical composition containing same Download PDF

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WO2021251756A1
WO2021251756A1 PCT/KR2021/007223 KR2021007223W WO2021251756A1 WO 2021251756 A1 WO2021251756 A1 WO 2021251756A1 KR 2021007223 W KR2021007223 W KR 2021007223W WO 2021251756 A1 WO2021251756 A1 WO 2021251756A1
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indol
chloro
methylene
carbohydrazide
ethoxyethyl
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French (fr)
Korean (ko)
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성낙균
김보연
황준성
한호진
이상구
한준열
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한국생명공학연구원
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/041—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0052—Small organic molecules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/041—Heterocyclic compounds
    • A61K51/0412—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K51/0419—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/041—Heterocyclic compounds
    • A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K51/0446—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • 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
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2123/00—Preparations for testing in vivo

Definitions

  • the present invention relates to a conjugate of an N-acylhydrazone derivative and a marker, and a pharmaceutical composition comprising the same.
  • the present invention relates to a conjugate of an N-acylhydrazone derivative excellent in targeting to cancer tissues and a ligand or fluorescent substance of a radioisotope, and a pharmaceutical composition for diagnosis of cell proliferative disease comprising the same.
  • the present invention relates to a novel N-acylhydrazone derivative and a pharmaceutical composition for preventing or treating a cell proliferative disease comprising the same.
  • a compound having an N-acylhydrazone skeleton is a promising drug from a pharmaceutical point of view, and research has been ongoing to improve it into a structure that specifically binds to cancer cells.
  • Korean Patent Application Laid-Open No. 2017-0098170 discloses that an N-acylhydrazone derivative having a specific structure having a benzofuran residue shows improved stability and solubility, and has an effect of inhibiting cell mitosis and cancer cell proliferation. is starting However, this document does not disclose the use of an N-acylhydrazone derivative as a diagnostic and therapeutic agent for cancer by conjugating it with a radioactive isotope ligand or a marker such as a fluorescent substance.
  • the present inventors observed a phenomenon in which N-acylhydrazone derivatives of a specific structure were specifically accumulated in cancer tissues in animals while researching the diagnosis and treatment of cancer in which a drug and a marker were conjugated. It was possible to develop a substance having an efficacy for diagnosing and treating an eating disorder.
  • an object of the present invention is to provide a novel N-acylhydrazone derivative-marker conjugate with excellent targeting to cancer, and a pharmaceutical composition useful for diagnosis and treatment of cancer, including the same.
  • Another object of the present invention is to provide an N-acylhydrazone derivative having a novel structure introduced into the conjugate, and a pharmaceutical composition containing the same with excellent anticancer effect.
  • the present invention provides a conjugate of an N-acylhydrazone derivative of Formula 1 and a marker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
  • R is , , or ego; A is -O- or -S-; R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl; R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl; R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy; R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino, and the marker is a ligand or a fluorescent substance of a radioisotope.
  • the present invention provides an N-acylhydrazone derivative represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  • the present invention provides a pharmaceutical composition for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising the conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof.
  • the present invention provides a pharmaceutical composition for preventing or treating a cell proliferative disease, comprising the conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof.
  • the present invention provides a pharmaceutical composition for preventing or treating cell proliferative diseases, comprising the N-acylhydrazone derivative, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer or a pharmaceutically acceptable salt thereof acts specifically on cancer cells and accumulates, and is prepared based on a compound having anticancer efficacy Therefore, both treatment and diagnosis are possible, and it has excellent stability and solubility.
  • N-acylhydrazone derivative of Formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same applied to the conjugate inhibit microtubule polymerization of tubulin to induce apoptosis. Therefore, it is useful for the prevention or treatment of cell proliferative diseases, including cancer.
  • 1 is a photograph confirming the effect of the concentration of an N-acylhydrazone derivative on the spindle and chromosomes of cells in mitosis by immunostaining.
  • Figures 2a to 2e show the results of testing the anticancer effect of the N-acylhydrazone derivative in a human cervical cancer cell (HeLa CCL2) transplantation model.
  • 3A to 3E are results of a stability test in plasma of an N-acylhydrazone derivative.
  • 4A to 4E are results of a metabolic stability test in liver microtubules of N-acylhydrazone derivatives.
  • 5a to 5c are pharmacokinetic (PK) test results of N-acylhydrazone derivatives.
  • 6A and 6B are results of a wound healing assay of N-acylhydrazone derivatives.
  • 7a and 7b are results of an in vitro invasion assay of N-acylhydrazone derivatives.
  • 11 and 12 show the results of measuring the targeting of the N-acylhydrazone derivative-phosphor conjugate to each tissue.
  • 15 is a result of measuring cancer tissue targeting of the N-acylhydrazone derivative-ligand conjugate over time.
  • 16 is a result of measuring the selective permeability of an N-acylhydrazone derivative-ligand conjugate to cancer cells.
  • halogen means F, Cl, Br or I, unless otherwise stated.
  • alkyl refers to a linear or branched saturated hydrocarbon moiety.
  • C 1-6 alkyl refers to alkyl having a backbone of 1 to 6 carbons. Specifically, C 1-6 alkyl is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, sec-pentyl, neopentyl , hexyl, and the like.
  • alkoxy refers to a linear or branched alkyl-oxy moiety.
  • C 1-6 alkoxy means alkyl-oxy with a backbone of 1 to 6 carbons. Specifically, C 1-6 alkoxy is methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, t-butoxy, n-pentoxy, i-pentoxy, t -pentoxy, sec-pentoxy, neopentoxy, hexyloxy, and the like may be included.
  • haloalkyl or haloalkoxy means an alkyl or alkoxy substituted with one or more halogens. Specifically, haloalkyl or haloalkoxy may be alkyl or alkoxy in which one or more homogeneous or heterogeneous halogens are substituted.
  • substitution refers to the replacement of a hydrogen atom in a molecular structure with a substituent such that the compound is chemically stable from such substitution without exceeding the valence on the designated atom.
  • group A is substituted with substituent B means that a hydrogen atom bonded to an atom such as carbon constituting the backbone of group A is replaced with substituent B, so that group A and substituent B form a covalent bond can do.
  • a novel N-acylhydrazone derivative-marker conjugate excellent in targeting to cancer, and an N-acylhydrazone derivative having a novel structure introduced into the conjugate are provided.
  • the N-acylhydrazone derivative has the general structure of Formula 1 below.
  • R is , , or ego; A is -O- or -S-; R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl; R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl; R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy; R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino.
  • the C 1-6 alkyl may include C 1-3 alkyl, C 3-6 alkyl, C 2-4 alkyl, C 2-6 alkyl, and the like.
  • the C 1-6 alkoxy may include C 1-3 alkoxy, C 3-6 alkoxy, C 2-4 alkoxy, C 2-6 alkoxy, and the like.
  • the haloC 1-6 alkyl and haloC 1-6 alkoxy may each have 1 to 10, or 1 to 3, identical or different halogens.
  • R is can be any organic radical
  • R is , , or can be
  • R 1 is H, -CH 3 , -CH 2 CO 2 CH 3 , -CH 2 CO 2 CH 2 CH 3 , -CH 2 CH 2 OCH 2 CH 3 or - CH 2 CH 2 OCH 3 .
  • R 2 and R 3 are each independently H, F, Cl, Br, -CH 3 -CF 3, -OCH 3 or -OCF 3.
  • R 4 and R 5 are each independently H, Cl, -CH 3, -OCH 3, or -NHCOCH 3.
  • R 1 is H, -CH 3 , -CH 2 CO 2 CH 3 , -CH 2 CO 2 CH 2 CH 3 , -CH 2 CH 2 OCH 2 CH 3 or - CH 2 CH 2 OCH 3 ;
  • R 2 is Cl, Br, —CH 3 or —CF 3 ;
  • R 3 is H, F, Cl, -CH 3 , -OCH 3, or -OCF 3, and
  • R 4 and R 5 are each independently H, Cl, -CH 3, -OCH 3, or -NHCOCH 3.
  • R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ;
  • R 2 is Cl, Br or —CH 3 ;
  • R 3 is H or —OCH 3 ;
  • R 4 and R 5 are each independently H, Cl, -CH 3 or -OCH 3 .
  • R is , , or ego;
  • R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ;
  • R 2 is Cl or —CH 3 ;
  • R 3 may be H or —OCH 3 .
  • N-acylhydrazone derivative of Formula 1 may include structures of Formulas 1a to 1e below.
  • R 1 , R 2 , R 3 , R 4 and R 5 are as exemplified in Formula 1 or an embodiment thereof.
  • R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl;
  • R 2 is halogen, C 1-6 alkyl or haloC 1-6 alkyl;
  • R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy;
  • R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino.
  • R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl; R 2 is halogen; R 3 is C 1-3 alkoxy.
  • R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl; R 2 is halogen; R 3 is H or C 1-3 alkoxy.
  • R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl.
  • R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl
  • R 2 is halogen or C 1-3 alkyl.
  • N-acylhydrazone derivative of Formula 1 may be selected from the group consisting of:
  • N-acylhydrazone derivative of Formula 1 may be selected from the group consisting of:
  • the N-acylhydrazone derivative of Formula 1 may be conjugated to a marker for diagnosis and effective treatment of cell proliferative diseases.
  • the marker may be a ligand of a radioisotope or a fluorescent substance.
  • Ligands of the radioactive isotopes include deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA).
  • DFO deferoxamine
  • DTPA diethylenetriaminepentaacetic acid
  • DAA 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid
  • the radioactive isotope may be at least one selected from the group consisting of 89 Zr, 60 Cu, 64 Cu, 223 Ra, 131 I, 89 Sr, 32 P, and 188 Re.
  • the phosphor may be at least one selected from the group consisting of cyanine 3, cyanine 5, cyanine 5.5, and cyanine 7, but is not limited thereto.
  • the ligand and the fluorescent substance are not limited thereto, and other ligands or fluorescent substances used for diagnosis and effective treatment of diseases are possible.
  • the N-acylhydrazone derivative is terminally conjugated with the marker.
  • it may be conjugated to the marker at the end of R 1 in Formula 1 above.
  • the conjugate may include a terminally aminated N-acylhydrazone derivative.
  • the terminal of R 1 of the N-acylhydrazone derivative may be aminated to be conjugated to the marker.
  • the N-acylhydrazone derivative may be conjugated to the marker by a linker.
  • the linker may include p-NCS-benzene.
  • the linker is not limited thereto, and other linkers used for conjugation are also possible.
  • the conjugate of the N-acylhydrazone derivative and the marker may be represented by the following formula (2):
  • R, R 2 , R 3 are as defined in Formula 1 above;
  • R 0 is a group in which one H is removed from R 1 in Formula 1;
  • L is a single bond or a group derived from a linker;
  • A is a group derived from a marker.
  • R 0 is a group derived from R 1 in Formula 1 above. That is, R 0 is a group (ie, a divalent radical group) in which an additional radical is formed by removing one H from R 1 (ie, a monovalent radical group) of Formula 1 above.
  • R 0 may be -(CH 2 ) m -, -(CH 2 ) n -COO-(CH 2 ) m -, or -(CH 2 ) n -O-(CH 2 ) m -, wherein m is an integer from 1 to 6, and n is an integer from 1 to 3. More specifically, -CH 2 , -CH 2 CO 2 CH 2 , -CH 2 CO 2 CH 2 CH 2 , -CH 2 CH 2 OCH 2 CH 2 or -CH 2 CH 2 OCH 2
  • L in Formula 2 may be a single bond. That is, the single bond of L may mean that it is directly bonded without a linker.
  • L in Formula 2 may be a group derived from a linker.
  • R, R 2 , and R 3 are as defined in Formula 1;
  • R 0 is a group in which one H is removed from R 1 in Formula 1;
  • L is a single bond or a group derived from a linker;
  • A is a group derived from a ligand or fluorescent substance of a radioisotope.
  • A may be a group derived from a ligand or a phosphor of a radioisotope.
  • A is a group derived from a ligand of a radioisotope, and may be a group in which one H or a terminal group is removed from the ligand to form a radical.
  • A is a group derived from deferoxamine (DFO)
  • DFO deferoxamine
  • A is a group derived from diethylenetriaminepentaacetic acid (DTPA)
  • DTPA diethylenetriaminepentaacetic acid
  • A is a group derived from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7,10-tetraazacyclododecane- One OH is removed from 1,4,7,10-tetraacetic acid, so that A may have the following structure.
  • DOTA 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid
  • A may have the following structure.
  • A is a group derived from a phosphor, and may be a group in which one H or a terminal group is removed from the phosphor to form a radical.
  • A is a group derived from cyanine 3 (Cy3)
  • Cy3 cyanine 3
  • A is a group derived from cyanine 5 (Cy5)
  • Cy5 the terminal group is removed from cyanine 5 so that A may have the following structure.
  • A is a group derived from cyanine 5.5 (Cy5.5)
  • Cy5.5 the terminal group is removed from cyanine 5.5, so that A may have the following structure.
  • A is a group derived from cyanine 7 (Cy7)
  • Cy7 a terminal group is removed from cyanine 7, so that A may have the following structure.
  • A in relation to the conjugate of the N-acylhydrazone derivative and the marker, in Formula 2, A may be any one selected from the groups listed above.
  • N-acylhydrazone derivative the N-acylhydrazone derivative defined in Formula 1 above can be easily applied to Formula 2, and a person skilled in the art can easily prepare a compound with reference to the following Examples.
  • N-acylhydrazone derivative As a specific example with respect to the conjugate of an N-acylhydrazone derivative and a marker, the following formulas 2a to 2g are described, and the N-acylhydrazone derivative can be easily substituted by replacing the compounds of the examples below. is not limited to
  • a conjugate represented by the following formula (2a), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
  • the conjugate represented by Formula 2a, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can be provided by conjugation via p-NCS-benzene with a linker. If necessary, a combined form without a linker may be provided.
  • a conjugate represented by Formula 2b, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a conjugate represented by Formula 2c, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a conjugate represented by Formula 2d, a stereoisomer or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a conjugate represented by Formula 2e, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a conjugate represented by Formula 2f, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a conjugate represented by Formula 2g, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
  • a method for preparing the conjugate of the N-acylhydrazone derivative and the marker is not particularly limited and may be carried out by a method suitable for each structure.
  • a conjugate may be prepared by obtaining an N-acylhydrazone derivative having an end amination in Chemical Formula 1 and reacting it with a marker.
  • a method for preparing a conjugate comprises the steps of (1) preparing an N-acylhydrazone derivative or a salt thereof having an terminal aminated in Formula 1; and (2) conjugating the aminated N-acylhydrazone derivative or salt thereof with a marker.
  • the terminal group in step (1), in order to prepare the terminally aminated N-acylhydrazone derivative or a salt thereof, the terminal group may be aminated after the synthesis procedure of the N-acylhydrazone derivative of Formula 1 above.
  • a substituent to be aminated in Formula 1 may be selected in advance, and a terminal of a reactant for introducing the substituent may be aminationed in advance, and then added to the synthesis reaction of the N-acylhydrazone derivative of Formula 1 above.
  • R, R 2 , R 3 are as defined in Formula 1 above;
  • R 0 is a group in which one H is removed from R 1 in Formula 1;
  • P is a protecting group;
  • X is halogen.
  • R 1 is reacted with an amine-protected halogenated amine compound, or (ii) R 1 is amination and After preparing a protected indole compound in advance, it is reacted with a hydrazone compound and then the protecting group is removed to obtain an N-acylhydrazone derivative in which the terminal of R 1 is aminated.
  • aminated N-acylhydrazone derivative may be further reacted with hydrochloric acid to form a salt, and then used in the next reaction to increase the final yield.
  • step (2) the conjugation of the aminated N-acylhydrazone derivative or salt thereof with the marker can be carried out in a conventional manner used in this field.
  • the conjugation may be performed through an amidation reaction or a thioamidation reaction, but is not limited thereto.
  • the conjugation can be carried out through a conventional linker, for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label.
  • a conventional linker for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label.
  • DFO deferoxamine
  • p-NCS-Bz-DFO a compound having p-NCS-benzene attached as a linker
  • the conjugation can be carried out through a conventional linker, for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label.
  • a conventional linker for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label.
  • DFO deferoxamine
  • p-NCS-Bz-DFO a compound to which p-NCS-benzene is attached as a linker may be added to the reaction.
  • the marker may use a modified terminus in order to increase the efficiency of the reaction.
  • cyanine 5 Cy5
  • NHS N-hydroxysuccinimide
  • the present invention includes an N-acylhydrazone derivative of Formula 1 or a pharmaceutically acceptable salt of the conjugate.
  • the pharmaceutically acceptable salt should have low toxicity to humans and should not adversely affect the biological activity and physicochemical properties of the parent compound.
  • the pharmaceutically acceptable salt may be an acid addition salt formed with a pharmaceutically acceptable free acid.
  • the free acid may be an inorganic acid or an organic acid, wherein the inorganic acid may be hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, hydrobromic acid, etc., and the organic acid is acetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid phonic acid, fumaric acid, maleic acid, malonic acid, phthalic acid, succinic acid, lactic acid, citric acid, gluconic acid, tartaric acid, salicylic acid, malic acid, oxalic acid, benzoic acid, embonic acid, aspartic acid, glutamic acid, and the like.
  • the inorganic acid may be hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, hydrobromic acid, etc.
  • the organic acid is acetic acid, methanesulfonic acid, ethane
  • the acid addition salt can be prepared by a conventional method, for example, by dissolving the parent compound in an excess aqueous acid solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. .
  • the pharmaceutically acceptable salt may be an alkali metal salt (sodium salt, etc.) or alkaline earth metal salt (potassium salt, etc.).
  • the alkali metal salt or alkaline earth metal salt can be obtained, for example, by dissolving the parent compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate.
  • N-acylhydrazone derivatives of formula (1) and the conjugates may have a chiral carbon center, and thus, in the form of R or S isomers, racemic compounds, individual enantiomers or mixtures, individual diastereomers or mixtures. may exist, and all such stereoisomers and mixtures thereof may fall within the scope of the present invention.
  • the present invention may include hydrates and solvates of the N-acylhydrazone derivative of Formula 1 or a conjugate thereof with a marker.
  • the hydrates and solvates can be prepared using a known method, and preferably nontoxic and water-soluble.
  • the hydrate and the solvate may be one in which 1 to 5 molecules of water and an alcoholic solvent (especially, ethanol, etc.) are bound, respectively.
  • the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer or a pharmaceutically acceptable salt thereof acts specifically on cancer cells and accumulates, and is prepared based on a compound having anticancer efficacy Therefore, both treatment and diagnosis are possible, and it has excellent stability and solubility.
  • N-acylhydrazone derivative of Formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same applied to the conjugate inhibit microtubule polymerization of tubulin to induce apoptosis. Therefore, it is useful for the prevention or treatment of cell proliferative diseases, including cancer.
  • the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof inhibits tubulin polymerization of microtubules to induce apoptosis and is also used in cancer cells exhibiting multidrug resistance. It works effectively and has excellent cancer metastasis inhibitory activity.
  • the compound according to the present invention exhibits high stability and solubility in the body, it has excellent bioavailability.
  • the compound according to the present invention or a pharmaceutical composition comprising the same can be used for diagnosis and prevention and treatment of cell proliferative diseases.
  • the compounds according to the invention can be used to depolymerize microtubules.
  • the compound according to the present invention can be used to inhibit tubulin, more specifically, to inhibit polymerization of tubulin.
  • the compound according to the present invention may act on the colchicine binding site of tubulin, and may induce apoptosis by stopping the cell cycle in the G2 or M phase.
  • the compound according to the present invention can act on cancer cells exhibiting multidrug resistance.
  • the compound according to the present invention or a pharmaceutical composition comprising the same can be used as a diagnostic agent for the occurrence, progression or metastasis of a cell proliferative disease.
  • the compound according to the present invention or a pharmaceutical composition comprising the same may be used as a contrast agent for the diagnosis of cell proliferative diseases.
  • the compound according to the present invention or a pharmaceutical composition comprising the same may be used as a tubulin inhibitor or an anticancer agent. Accordingly, the present invention provides the use of a compound according to the present invention for diagnosing the occurrence, progression or metastasis of a cell proliferative disease.
  • the present invention also provides the use of the compound according to the present invention for the prevention or treatment of a cell proliferative disease.
  • the invention also provides the use of a compound according to the invention for inhibiting polymerization of tubulin.
  • the present invention also provides the use of a compound according to the present invention for the manufacture of a medicament for diagnosis of the occurrence, progression or metastasis of a cell proliferative disease.
  • the present invention also provides a composition comprising a compound according to the present invention for use in the diagnosis of the development, progression or metastasis of a cell proliferative disease.
  • the present invention also provides the use of the compound according to the present invention for the manufacture of a contrast agent for diagnosis of a cell proliferative disease.
  • the present invention also provides the use of a compound according to the present invention for the manufacture of a medicament for diagnosis of the occurrence, progression or metastasis of a cell proliferative disease.
  • the present invention also provides the use of the compound according to the present invention for the preparation of a medicament for preventing or treating a cell proliferative disease.
  • the present invention also provides the use of the compound according to the present invention for the preparation of a medicament for inhibiting tubulin polymerization.
  • the present invention also provides a method for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof.
  • the present invention also provides a method of imaging a site having a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof.
  • the present invention also provides a method for inhibiting polymerization of tubulin comprising administering to a subject in need thereof a compound according to the present invention.
  • the present invention also provides a method for preventing or treating a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof.
  • diagnosis refers to any act of discovering and detecting the disease by administration of the compound or imaging using imaging equipment
  • prevention refers to any activity that inhibits the occurrence, spread and recurrence of the disease by administration of the compound. It means any action that delays or improves
  • treatment means any action in which the symptoms of the disease are ameliorated or beneficially changed by administration of the compound.
  • the term "subject in need” refers to monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, including humans (patients) who have or may develop the cell proliferative disease. It refers to all animals such as rabbits and guinea pigs, and may specifically refer to mammals. In addition, the required object may refer to a biological sample.
  • administration means providing a predetermined substance to a subject in need thereof by any suitable method, and the administration route of the compound according to the present invention is administered through any general route as long as it can reach the target tissue.
  • the present invention also provides a pharmaceutical composition comprising the compound according to the present invention as an active ingredient.
  • the present invention also provides a pharmaceutical composition for inhibiting polymerization of tubulin comprising the compound according to the present invention as an active ingredient.
  • the present invention also provides a pharmaceutical composition, specifically, a contrast agent for diagnosing the occurrence, progression or metastasis of a cell proliferative disease comprising the compound according to the present invention as an active ingredient.
  • the pharmaceutical composition may also exhibit a therapeutic effect on cell proliferative diseases.
  • the present invention also provides a pharmaceutical composition for the prevention or treatment of a cell proliferative disease comprising the compound according to the present invention as an active ingredient.
  • the cell proliferative disease may be cancer.
  • cancer refers to the abnormal growth of cells that tend to proliferate or metastasize in an uncontrolled manner.
  • the cancer may be a solid cancer, a blood cancer, or a metastatic cancer. More specifically, the cancer may be rectal cancer, breast cancer, lung cancer, stomach cancer, liver cancer, leukemia, glioma, skin cancer, cervical cancer, or a metastasis derived therefrom.
  • the pharmaceutical composition comprising the compound according to the present invention can be used as a diagnostic agent, contrast agent, therapeutic agent, or cancer metastasis inhibitor for various cancers exemplified above.
  • the present invention also provides a pharmaceutical composition comprising the compound according to the present invention and a pharmaceutically acceptable additive.
  • the pharmaceutical composition of the present invention contains the compound according to the present invention as an active ingredient, 0.1 wt% to 90 wt%, specifically 0.1 wt% to 75 wt%, more specifically 1 wt%, based on the total weight of the pharmaceutical composition to 50% by weight.
  • the pharmaceutical composition of the present invention may include conventional and non-toxic pharmaceutically acceptable excipients according to conventional methods.
  • the pharmaceutical composition may further include a pharmaceutically acceptable carrier, diluent or excipient.
  • additives used in the pharmaceutical composition of the present invention may include sweeteners, binders, solvents, solubilizing agents, wetting agents, emulsifiers, isotonic agents, absorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, flavoring agents, and the like. have.
  • the additive may be lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, stearin, magnesium stearate, magnesium aluminosilicate, starch, gelatin, gum tragacanth, alginic acid, sodium alginate, methylcellulose, sodium carboxymethylcellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavor, and the like.
  • composition of the present invention may be formulated in various formulations for oral administration (eg, tablets, pills, powders, capsules, syrups or emulsions) or parenteral administration (eg, intramuscular, intravenous or subcutaneous injection).
  • oral administration eg, tablets, pills, powders, capsules, syrups or emulsions
  • parenteral administration eg, intramuscular, intravenous or subcutaneous injection.
  • the pharmaceutical composition of the present invention may be formulated as a formulation for oral administration, and the additives used in this case include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, and magnesium stearate. lactate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifying agents, diluents, and the like may be included.
  • solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the pharmaceutical composition, for example, starch, calcium carbonate, sucrose, It may be formulated by mixing lactose, gelatin, and the like.
  • excipients for example, starch, calcium carbonate, sucrose, It may be formulated by mixing lactose, gelatin, and the like.
  • lubricants such as magnesium stearate and talc may be used.
  • liquid formulations for oral administration may be exemplified by suspensions, emulsions, syrups, etc., and various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are commonly used simple diluents. may be included.
  • preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories.
  • Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.
  • the suppositories are Witepsol, Macrogol, and Twin61. Cacao butter, laurin fat, glycerogelatin, etc. may be used.
  • the injection may contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, and the like.
  • the pharmaceutical composition when used for the diagnosis of cell proliferative diseases, for example, as a fluorescent contrast agent, the compound according to the present invention may be suspended or dissolved in a solvent such as distilled water for injection, pharmacological saline, Ringer's solution, etc. can If necessary, pharmaceutically acceptable additives such as carriers, excipients and the like may be added.
  • pharmaceutically acceptable additives such as carriers, excipients and the like may be added.
  • Such additives include pharmacologically acceptable electrolytes, buffers, detergents and substances for regulating osmotic pressure and substances for improving stability and solubility (eg, cyclodextrins, liposomes, etc.).
  • various additives commonly used in the related field may be used.
  • the pharmaceutical composition when used as a radioactive contrast agent for radiodiagnosis, for example, proton emission tomography (PET), it may be formulated as an injection, and more specifically, it may be formulated as an intravenous injection.
  • the pharmaceutical composition may include a non-toxic buffer solution isotonic with blood as a diluent, and may include, for example, a phosphate buffer solution of pH 7.4, and other diluents or additives in addition to the buffer solution. may include more. Excipients and additives that can be added to such injections are well known to those of ordinary skill in the art.
  • the compound or pharmaceutical composition according to the present invention may be administered to a patient in a therapeutically effective amount or in a pharmaceutically effective amount.
  • terapéuticaally effective amount refers to an amount of a compound or pharmaceutical composition effective to prevent or treat a target disease, and is used to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment. It is sufficient and refers to an amount sufficient to not cause side effects.
  • the level of the effective amount may be determined by the patient's health condition, disease type, severity, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment period, factors including the combination or concurrently used drugs; It may be determined according to factors well known in the medical field.
  • the "pharmaceutically effective amount” may refer to an amount of a compound or pharmaceutical composition effective for diagnosing a target disease.
  • the compound or pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.
  • the effective amount of the compound in the pharmaceutical composition of the present invention may vary depending on the age, sex, and weight of the patient, and in general, 0.1 mg to 100 mg, or 0.5 mg to 10 mg per kg of body weight, daily or every other day It can be administered or divided into 2 or 3 times a day.
  • the scope of the present invention is not limited thereto since it may increase or decrease depending on the route of administration, disease severity, sex, weight, age, etc.
  • the pharmaceutical composition of the present invention when used as a fluorescent contrast agent, it can be administered to a living body by injection, spraying or application, intravenous (venous, arterial), oral, intraperitoneal, transdermal, subcutaneous, intracystic or intrabronchial.
  • the dosage of the pharmaceutical composition is 0.1 mg to 100 mg, preferably 0.5 mg to 20 mg per 1 kg of body weight, in the amount of the compound according to the present invention, the type of compound emitting fluorescence used, the age of the subject, It can be appropriately adjusted according to body weight and target organs.
  • the pharmaceutical composition of the present invention when used for radiodiagnosis, for example, as a PET contrast agent, an amount of about 1 mCi to 3 mCi is administered based on a human male adult based on the compound according to the present invention as an active ingredient.
  • a professional doctor can appropriately increase or decrease it.
  • the compound or pharmaceutical composition according to the present invention may be administered for tumor therapy in combination with chemotherapy, radiation therapy, immunotherapy, hormone therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapy, surgical intervention or a combination thereof.
  • the compound or pharmaceutical composition according to the present invention can be used as an adjuvant therapy in combination with other long-term treatment strategies, or to maintain the patient's condition after tumor regression or chemopreventive therapy in seriously ill patients.
  • the pharmaceutical composition of the present invention may further include one or more active ingredients, wherein the additional active ingredient is an anti-proliferative compound, such as an aromatase inhibitor, anti-estrogens, topoisomerase I Inhibitors, topoisomerase II inhibitors, microtubule active compounds, alkylation compounds, histone deacetylase inhibitors, compounds inducing cellular differentiation processes, cyclooxygenase inhibitors, MMP inhibitors, mTOR inhibitors, anti-neoplastic anti-metabolism Substances, platin compounds, compounds targeting/reducing protein or lipid kinase activity, anti-angiogenic compounds, compounds targeting, reducing or inhibiting the activity of protein or lipid phosphatase, gonadorelin agonists, anti-androgens, Methionine aminopeptidase inhibitors, bisphosphonates, biological response modifiers, anti-proliferative antibodies, heparanase inhibitors, inhibitors of Ras tumorigenic isoforms,
  • PrOH propanol
  • DMSO dimethyl sulfoxide
  • Ethyl 5-aminobenzofuran-2-carboxylate (205.2 mg, 1.0 mmol) was dissolved in DMF (3 mL), and then (2-(1H-benzotriazol-1-yl)-1,1,3 ,3-Tetramethyluronium hexafluorophosphate (HBTU, 417.2 mg, 1.1 mmol), hydroxybenzotriazole (HOBt, 148.6 mg, 1.1 mmol), N,N-diisopropylethylamine (DIEA, 209.0 ⁇ L) , 1.2 mmol), and acetic acid (63.0 ⁇ L, insert the 1.1 mmol) was 15 hours at room temperature.
  • HBTU (2-(1H-benzotriazol-1-yl)-1,1,3 ,3-Tetramethyluronium hexafluorophosphate
  • HOBt hydroxybenzotriazole
  • DIEA N,N-diisopropy
  • Benzo[b]thiophene-2-carboxylic acid (100 mg, 0.56 mmol, Combi-blocks) was dissolved in ethanol (2 mL), concentrated sulfuric acid (1-2 drops) was added, and the mixture was heated and refluxed for 10 hours. Ethanol was concentrated under reduced pressure, and the residue was dissolved in EA , washed with saturated NaHCO 3 aqueous solution, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was dissolved in ethanol (2 mL), hydrazine monohydrate (84 mg, 1.68 mmol) was added, and refluxed for 20 hours. The reaction solution was cooled and the resulting solid was filtered, washed with ethyl ether and dried to obtain the title compound (100 mg, 93%).
  • the title compound (yield 90%, yellow solid) was obtained by the synthesis method disclosed in Korean Patent Registration No. 1790193.
  • the title compound (yield 75%, light yellow solid) was obtained by the synthesis method disclosed in Korean Patent Application Laid-Open No. 2014-0128238.
  • N-acylhydrazone derivative-labeler conjugate was prepared based on the compounds of Examples above.
  • the preparation methods of Examples 29 and 30 below show an example of the preparation of the N-acylhydrazone derivative-labeler conjugate in the case of conjugation via p-NCS-benzene and the case of direct conjugation without a linker.
  • a person skilled in the art can proceed with synthesis according to appropriate synthesis conditions using the above N-acylhydrazone derivatives together with commercially available compounds (radioactive isotope ligands or fluorescent substances, etc.).
  • Step 1 Preparation of t-butyl (2-(2-hydroxyethoxy)ethyl)carbamate
  • Step 2 Preparation of t-butyl (2-(2-bromoethoxy)ethyl)carbamate
  • Step 3 Preparation of t-butyl (2-(2-(2-chloro-3-formyl-1H-indol-1-yl)ethoxy)ethyl)carbamate
  • Step 6 t-Butyl (E)-(2-(2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl)-1H-indole- Preparation of 1-yl) ethoxy) ethyl) carbamate
  • Step 7 (E)-N'-((1-(2-(2-aminoethoxy)ethyl)-2-chloro-1H-indol-3-yl)methylene)-5-methylbenzofuran-2- Preparation of Carbohydrazide Hydrochloride
  • Step 8 (E)-N 1 -(5-(3-(4-(3-(2-(2-(2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl) )hydrazinylidene)methyl)-1H-indol-1-yl)ethoxy)ethyl)thioureido)phenyl)thioureido)pentyl)-N 1 -hydroxy-N 4 -(5-(N) Preparation of -hydroxy-4-((5-(N-hydroxyacetamido)pentyl)amino)-4-oxobutanamido)pentyl)succinamide
  • Example 4 - MRM the compound of Example 4 - m/z 424>159 (Ret. Time: 3.8 min), carbamazepine (IS) - m/z 237>194 (Ret. Time: 3.2 min)
  • HeLa cells ATCC, USA
  • a human cervical cancer cell line were seeded in a 96-well plate at 3 x 10 3 cells/well, and then the cells were treated with the compounds of Examples according to the present invention and grown for 2 days.
  • MTT 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
  • absorbance was measured at "OD 450" and statistical values were obtained using Prism TM 6 program.
  • Table 3 are average values of the results of repeating the analysis twice.
  • Example compound according to the present invention exhibits anti-proliferative activity not only in the HeLa cell line (cervical cancer cell line) but also in other cancer cells.
  • Example compound according to the present invention has an effect on cancer cell lines showing multi-drug resistance.
  • K562 and MCF7 Bio Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Korea
  • K562/ADR and MCF7/ADR Bio Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Korea
  • IC 50 was obtained by a log-dose response curve (unit: nM).
  • the data shown in the table below are the average values of the results obtained by repeating the analysis three times.
  • Resistance factors (resistance factor) of a cell line exhibit multidrug resistance means the ratio of the IC 50 of the multi-drug-resistant cell line to the IC 50 of the primary (parent) cells with no resistance.
  • the multi-drug-resistant cell line exhibited great resistance, tens to hundreds of times the resistance factor to the existing anticancer drugs.
  • the resistance factor was 0.40 to 0.57, indicating that the example compound according to the present invention exhibits a stronger cytotoxic effect on cancer cell lines showing multidrug resistance compared to the existing anticancer drugs.
  • the HeLa CCL2 cell line was cultured in a 12-well plate (3x10 4 cells/well), treated with DMSO or the compound of Example 4 for 17 hours, and then propidium iodine dye was added to stain the cell DNA and FACS. was used.
  • the cell concentration and the number of cells in the dividing phase are expressed as a percentage.
  • the compound of Example 4 showed an IC 50 at 0.2 ⁇ M against HeLa CCL2, a representative cancer cell line, and in particular, more than 80% of the cells stopped in the mitotic phase after 16 hours after treatment with the cells. It was expected to inhibit the polymerization reaction of tubulin by observing that there is, and this was confirmed in Experimental Example 3.
  • HeLa cells were treated with DMSO or the compound of Example 4 (50 nM, 100 nM, 200 nM) for 16 hours.
  • the cells After fixing the cells, they were stained with an anti-tubulin antibody and Alexa Fluor TM 488, and nuclei of the cells were immunostained using Hoechst 33342 to test ⁇ -tubulin and DNA. As shown in FIG. 1 , the shape of tubulin became coarser and shorter as the concentration increased when treated with the example compound according to the present invention compared to the DMSO control group. In addition, the form of DNA deviating from the central alignment also increased.
  • Example compound according to the present invention is an agent for depolymerizing microtubules.
  • Example compounds according to the present invention were used as test substances, and solvents (carriers) were used as negative controls.
  • the compounds were dissolved in a mixture of DMAC (dimethylacetamide) 20% + Tween 80 5% + 20% HPbCD (2-hydroxypropyl- ⁇ -cyclodextrin) 75% to an appropriate concentration before use.
  • DMAC dimethylacetamide
  • HPbCD 2-hydroxypropyl- ⁇ -cyclodextrin
  • the tumor size for each animal was measured 11 times in 3 directions using a vernier caliper, and then length x width x height / 2 It was calculated by the formula of
  • Example compound In order to test the degree of toxicity upon repeated intraperitoneal administration of the Example compound to HeLa CCL2 cancer cell transplantation nude mice, general symptoms and body weight changes of the animals were observed during the administration period.
  • tumor weight reductions of 57.1%, 40.5%, and 26.2% were observed in the groups administered with the compounds of Examples 4, 8 and 15, respectively (see FIGS. 2D and 2E ).
  • the compounds of Examples 2, 3, 4, 8 and 15 were administered intravenously, and plasma concentrations over time were observed. Also, Procaine was used as a positive control. Plasma stability was measured three times by administering the test compound at a concentration of 5 mM to human, rat or mouse plasma (90 ⁇ L) at 37°C. The results are shown in Table 8 and FIGS. 3A to 3E below.
  • test compound was administered at a concentration of 1 mM in the presence or absence of NADPH (1 mM) to liver microtubules (0.5 mg protein/mL) of humans, rats and mice, and metabolic stability was tested at 37° C. for 30 minutes. Also, Buspirone was used as a positive control. The results are shown in Tables 9 and 10 below; and FIGS. 4A to 4E .
  • Example 3 human +NADPH 8.69 ⁇ 0.44 12.34 ⁇ 0.69 15.76 ⁇ 0.89 -NADPH 36.40 ⁇ 1.90 22.26 ⁇ 0.72 77.60 ⁇ 1.39 rat +NADPH 2.99 ⁇ 0.64 1.39 ⁇ 0.12 3.23 ⁇ 0.55 -NADPH 41.45 ⁇ 0.93 8.53 ⁇ 0.46 69.63 ⁇ 1.94 mouse +NADPH 2.84 ⁇ 0.34 2.50 ⁇ 0.11 5.48 ⁇ 0.04 -NADPH 11.59 ⁇ 1.06 6.81 ⁇ 0.17 47.95 ⁇ 2.93 buffer 77.94 ⁇ 7.04 89.02 ⁇ 4.39 100.93 ⁇ 11.79
  • Example 15 buspirone human +NADPH 24.11 ⁇ 0.46 7.94 ⁇ 0.74 2.21 ⁇ 0.03 -NADPH 99.18 ⁇ 1.92 88.63 ⁇ 1.61 91.06 ⁇ 0.40 rat +NADPH 4.55 ⁇ 0.94 3.47 ⁇ 0.25 0.41 ⁇ 0.02 -NADPH 85.58 ⁇ 3.34 94.95 ⁇ 1.93 93.08 ⁇ 1.21 mouse +NADPH 11.41 ⁇ 2.47 4.82 ⁇ 0.52 0.32 ⁇ 0.05 -NADPH 89.93 ⁇ 6.72 86.31 ⁇ 1.72 94.74 ⁇ 0.74 buffer 98.98 ⁇ 2.07 91.02 ⁇ 2.89 -
  • Solvent B Cremophor EL, SigmaAldrich
  • Solvent C 20% HPbCD ((2-hydroxypropyl)- ⁇ -cyclodextrin, SigmaAldrich) in deionized water
  • PK pharmacokinetic
  • PK Pharmacokinetic
  • Example 8 parameter Intravenous, 5 mg/kg Oral, 20 mg/kg average Standard Deviation average Standard Deviation t max hr N/A 0.58 0.14 C max ng/mL N/A 93.87 5.19 AUC last ng*hr/mL 2377.94 112.06 96.27 9.58 AUC inf ng*hr/mL 2386.74 112.22 102.54 8.2 CL L/hr/kg 2.10 0.10 195.92 16.1 V ss L/kg 0.50 0.03 N/A V z L/kg 0.85 0.05 N/A t 1/2 hr 0.28 0.01 0.4 0.0 MRT inf hr 0.24 0.00 0.9 0.1 F % N/A 1.0 0.10
  • Example 15 parameter IV 5 mg/kg PO, 20 mg/kg average Standard Deviation average Standard Deviation t max hr N/A 0.25 0.00 C max ng/mL N/A 34.43 27.34 AUC last ng*hr/mL 1717.55 41.34 18.00 18.40 AUC inf ng*hr/mL 1729.14 41.24 NC NC CL L/hr/kg 2.89 0.07 NC NC V ss L/kg 0.71 0.02 N/A V z L/kg 1.37 0.09 N/A t 1/2 hr 0.33 0.02 NC NC MRT inf hr 0.24 0.01 NC NC NC F % N/A 0.3 0.27
  • Example compounds had stability that could be used as drugs.
  • HeLa CCL2 was cultured in 6 wells at 100%, and after applying a wound with a yellow tip, the compound of Example 4 was treated with 50 nM and 100 nM.
  • the state of the cells before treatment was recorded as a photograph, and compared with the state of the cells at 24, 36, and 48 hours after the drug treatment, the degree of cell migration was comparatively observed.
  • the concentration of the compound of Example 4 treated with the cells was determined by performing MTT.
  • the concentration of the compound of Example 4 did not have a significant effect on cell growth up to 100 nM.
  • an invasion assay was performed. Prepare a trans well chamber for a 24-well plate , put 5 X 10 4 of HeLa CCL2 in the upper chamber, and process the compound of Example 4 10 nM, 50 nM, 100 nM, 250 nM After incubation, after 16 hours, the migrated cells were identified and the number was measured through cell staining.
  • Example 4 As a result, as shown in FIGS. 7a and 7b, it was confirmed that the compound of Example 4 according to the present invention had an excellent cancer metastasis inhibitory effect.
  • N-acylhydrazone derivative-phosphor conjugate of Example 30 Targeting of the N-acylhydrazone derivative-phosphor conjugate of Example 30 to cancer tissue was evaluated as a drug-phosphor conjugate.
  • the solution from the 4th to 5th tube had the highest purity, and as it went further back, the tube contained more 89 Zr 4+ that could not be combined with the conjugate.
  • the tube with the highest radioactivity value it was intravenously injected into the tail of a xenograft mouse model at a maximum of 200 ⁇ Ci and a minimum of 50 ⁇ Ci (as the radioactivity value varies for each junction, all mice were treated when the injection amount was determined. were injected with the same radioactive dose).
  • mice were sacrificed and organs were removed, followed by in vitro experiments. Mice were sacrificed at 1, 24, 48, 72 and 96 hours at the same time as PET imaging, and quantitative values of each organ were measured with a gamma counter (Wizard2, PerkinElmer). As shown in FIG. 15 , the conjugate of Example 29 was measured at a high level in the tumor. Through these results, it was confirmed that the drug-conjugate ligand accurately targets the tumor, and the same results as the previous experiment, the drug-fluorescent conjugate, were shown, confirming once more that the drug tumor targeting experiment was significant. That is, the excellent effect on tumor targeting of the conjugate of the N-acylhydrazone derivative and the marker according to the present invention was confirmed.
  • the N-acylhydrazone-ligand conjugate of Example 29 conjugate of the compound of Example 4 with DFO
  • 1x10 5 HeLa CCL2 was cultured in a 12-well plate for 24 hours, and the medium was removed 1, 2, and 4 hours after the drug-ligand conjugate was administered alone or simultaneously with a competing drug, and then the cancer cells were washed with PBS. After dissolving cancer cells with 1 N NaOH, it was placed in a transparent test tube and quantitative values were measured with a gamma meter.

Abstract

According to the present invention, a conjugate of an N-acylhydrazone derivative of chemical formula 1 and a marker, or a stereoisomer or pharmaceutically acceptable salt thereof specifically acts on cancer cells and accumulates, and is prepared on the basis of a compound having anticancer efficacy so that both treatment and diagnosis are possible and has excellent stability and solubility.

Description

N-아실히드라존 유도체-표지자 접합체 및 이를 포함하는 약학적 조성물N-acylhydrazone derivative-marker conjugate and pharmaceutical composition comprising same
본 발명은 N-아실히드라존 유도체와 표지자의 접합체 및 이를 포함하는 약학적 조성물에 관한 것이다. 구체적으로, 본 발명은 암조직에 타겟팅이 우수한 N-아실히드라존 유도체와 방사성 동위원소의 리간드 또는 형광체의 접합체, 및 이를 포함하는 세포증식성 질환의 진단용 약학적 조성물에 관한 것이다. 또한 본 발명은 신규의 N-아실히드라존 유도체 및 이를 포함하는 세포증식성 질환의 예방 또는 치료용 약학적 조성물에 관한 것이다. The present invention relates to a conjugate of an N-acylhydrazone derivative and a marker, and a pharmaceutical composition comprising the same. Specifically, the present invention relates to a conjugate of an N-acylhydrazone derivative excellent in targeting to cancer tissues and a ligand or fluorescent substance of a radioisotope, and a pharmaceutical composition for diagnosis of cell proliferative disease comprising the same. In addition, the present invention relates to a novel N-acylhydrazone derivative and a pharmaceutical composition for preventing or treating a cell proliferative disease comprising the same.
국내 암환자의 24%가 방사선 치료를 받고 있으며, 방사선 치료를 받는 환자수는 6.2%씩 증가하고 있다. 특히, 암의 조기 진단을 통해 환자의 예후가 좋아지고 생존 및 치료의 가능성이 높아짐에 따라 암 진단에 대한 관심이 증가하고 있다. 방사선을 이용한 의료영상 장비는 인체 내부를 비침습적으로 영상화할 수 있기 때문에 많은 병원 및 의료기관에서 질병의 진단과 치료 후 경과 관찰 등을 위해 폭넓게 사용되고 있다. 특히 암 진단 시에는 핵의학 방사성 의약품을 투여하여 인체 내부에서 투과되어 나오는 감마선을 검출해 종양의 위치정보 및 그 외의 생리학적 정보를 얻어내는 방식이 많이 활용되고 있다. 이러한 진단용 방사성 의약품은 크게 단일광자방사형 컴퓨터단층촬영(SPECT)용과 양전자방출 단층촬용(PET)용으로 나눌 수 있는데, SPECT용 방사성 의약품은 주로 Tc-99m를 표지하여 만들고, 2018년 기준으로 전세계 SPECT용 방사성의약품 시장의 88% 가량을 차지하고 있다.24% of cancer patients in Korea are receiving radiation therapy, and the number of patients receiving radiation therapy is increasing by 6.2%. In particular, interest in cancer diagnosis is increasing as the prognosis of patients is improved through early diagnosis of cancer and the possibility of survival and treatment increases. Since medical imaging equipment using radiation can image the inside of the human body non-invasively, it is widely used in many hospitals and medical institutions to diagnose diseases and observe progress after treatment. In particular, in cancer diagnosis, nuclear medicine radiopharmaceuticals are administered to detect gamma rays penetrating from the inside of the human body to obtain tumor location information and other physiological information. These diagnostic radiopharmaceuticals can be largely divided into single photon emission computed tomography (SPECT) and positron emission tomography (PET). It accounts for about 88% of the radiopharmaceutical market.
기존의 암 진단용 물질은 주로 항체를 중심으로 한 단백질류로서 체내에 적용이 용이하지 못하고 생산단가가 높은 단점이 있었다. 이에 암세포에 특이적인 약물을 방사성 동위원소의 리간드 또는 형광체와 같은 표지자와 접합하여 암의 진단 및 치료제로 개발하기 위한 노력이 이어지고 있다. 이러한 암의 진단 및 치료에 사용되는 약물은 안정성 및 용해도가 높고 암세포에 특이적으로 축적되며 나아가 암세포 증식 억제 효과를 나타내는 것이 필요하다. Existing cancer diagnostic materials are mainly proteins centered on antibodies, and have disadvantages in that they cannot be easily applied to the body and the production cost is high. Accordingly, efforts are being made to develop cancer diagnosis and therapeutic agents by conjugating a cancer cell-specific drug with a radioisotope ligand or a marker such as a fluorescent substance. Drugs used for diagnosis and treatment of these cancers need to have high stability and solubility, to accumulate specifically in cancer cells, and to further exhibit an inhibitory effect on cancer cell proliferation.
N-아실히드라존 골격을 갖는 화합물은 약제학적 관점에서 유망한 약물로서, 암세포에 특이적으로 결합하는 구조로 개량하기 위한 연구가 계속되어 왔다. 예를 들어 한국 공개특허공보 제 2017-0098170 호는 벤조퓨란계 잔기를 갖는 특정 구조의 N-아실히드라존 유도체가 개선된 안정성 및 용해도를 나타내고 세포 유사분열과 암세포의 증식을 억제하는 효과를 가짐을 개시하고 있다. 그러나 상기 문헌은 N-아실히드라존 유도체를 방사성 동위원소의 리간드 또는 형광체와 같은 표지자와 접합하여 암의 진단 및 치료제로 사용하는 것에 대해서는 개시하지 않고 있다.A compound having an N-acylhydrazone skeleton is a promising drug from a pharmaceutical point of view, and research has been ongoing to improve it into a structure that specifically binds to cancer cells. For example, Korean Patent Application Laid-Open No. 2017-0098170 discloses that an N-acylhydrazone derivative having a specific structure having a benzofuran residue shows improved stability and solubility, and has an effect of inhibiting cell mitosis and cancer cell proliferation. is starting However, this document does not disclose the use of an N-acylhydrazone derivative as a diagnostic and therapeutic agent for cancer by conjugating it with a radioactive isotope ligand or a marker such as a fluorescent substance.
본 발명자들은 약물과 표지자를 접합시킨 암의 진단 및 치료제를 연구하던 중, 특정 구조의 N-아실히드라존 유도체가 동물에서 암조직에 특이적으로 축적되는 현상을 관찰하였고, 이를 토대로 암과 같은 세포증식성 질환을 진단 및 치료할 수 있는 효능을 갖는 물질을 개발할 수 있었다.The present inventors observed a phenomenon in which N-acylhydrazone derivatives of a specific structure were specifically accumulated in cancer tissues in animals while researching the diagnosis and treatment of cancer in which a drug and a marker were conjugated. It was possible to develop a substance having an efficacy for diagnosing and treating an eating disorder.
따라서 본 발명은 암에 대한 타겟팅이 뛰어난 새로운 N-아실히드라존 유도체-표지자 접합체, 및 이를 포함하여 암의 진단 및 치료에 유용한 약학적 조성물을 제공하고자 한다. 또한 본 발명은 상기 접합체에 도입되는 새로운 구조의 N-아실히드라존 유도체, 및 이를 포함하는 우수한 항암 효과의 약학적 조성물을 제공하고자 한다.Accordingly, an object of the present invention is to provide a novel N-acylhydrazone derivative-marker conjugate with excellent targeting to cancer, and a pharmaceutical composition useful for diagnosis and treatment of cancer, including the same. Another object of the present invention is to provide an N-acylhydrazone derivative having a novel structure introduced into the conjugate, and a pharmaceutical composition containing the same with excellent anticancer effect.
상기 과제에 따라, 본 발명은 한 측면에서 하기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공한다:According to the above object, in one aspect, the present invention provides a conjugate of an N-acylhydrazone derivative of Formula 1 and a marker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[화학식 1][Formula 1]
Figure PCTKR2021007223-appb-img-000001
Figure PCTKR2021007223-appb-img-000001
상기 식에서 R은
Figure PCTKR2021007223-appb-img-000002
,
Figure PCTKR2021007223-appb-img-000003
,
Figure PCTKR2021007223-appb-img-000004
또는
Figure PCTKR2021007223-appb-img-000005
이고; A는 -O- 또는 -S-이고; R1은 H, C1-6알킬, C1-6알콕시카보닐C1-3알킬, 또는 C1-6알콕시C1-3알킬이고; R2는 할로겐, C1-6알킬 또는 할로C1-6알킬이고; R3은 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 할로C1-6알콕시이고; R4 및 R5는 각각 독립적으로 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 C1-6알킬카보닐아미노이고, 상기 표지자는 방사성 동위원소의 리간드 또는 형광체이다.where R is
Figure PCTKR2021007223-appb-img-000002
,
Figure PCTKR2021007223-appb-img-000003
,
Figure PCTKR2021007223-appb-img-000004
or
Figure PCTKR2021007223-appb-img-000005
ego; A is -O- or -S-; R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl; R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl; R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy; R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino, and the marker is a ligand or a fluorescent substance of a radioisotope.
본 발명은 다른 측면에서 상기 화학식 1로 표시되는 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공한다.In another aspect, the present invention provides an N-acylhydrazone derivative represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
본 발명은 또 다른 측면에서 상기 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 포함하는, 세포증식성 질환의 발생, 진행 또는 전이를 진단하기 위한 약학적 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising the conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof.
본 발명은 또 다른 측면에서 상기 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 포함하는, 세포증식성 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition for preventing or treating a cell proliferative disease, comprising the conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof.
본 발명은 또 다른 측면에서 상기 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는, 세포증식성 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition for preventing or treating cell proliferative diseases, comprising the N-acylhydrazone derivative, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명에 따른 상기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염은 암세포에 특이적으로 작용하여 축적되며, 항암 효능을 갖는 화합물을 기반으로 제조되어 치료와 진단이 모두 가능하고 안정성과 용해도가 우수하다.According to the present invention, the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer or a pharmaceutically acceptable salt thereof, acts specifically on cancer cells and accumulates, and is prepared based on a compound having anticancer efficacy Therefore, both treatment and diagnosis are possible, and it has excellent stability and solubility.
또한 상기 접합체에 적용되는 상기 화학식 1의 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염, 및 이를 포함하는 약학적 조성물은 미세소관의 튜불린 중합을 저해하여 세포자살을 유발하므로 암을 비롯한 세포증식성 질환의 예방 또는 치료에 유용하다.In addition, the N-acylhydrazone derivative of Formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same applied to the conjugate inhibit microtubule polymerization of tubulin to induce apoptosis. Therefore, it is useful for the prevention or treatment of cell proliferative diseases, including cancer.
도 1은 N-아실히드라존 유도체의 농도에 따라 유사분열 중인 세포의 방추사와 염색체에 미치는 영향을 면역 염색으로 확인한 사진이다.1 is a photograph confirming the effect of the concentration of an N-acylhydrazone derivative on the spindle and chromosomes of cells in mitosis by immunostaining.
도 2a 내지 2e는 N-아실히드라존 유도체의 인체 유래 자궁경부암 세포(HeLa CCL2) 이식 모델에서의 항암 효과 시험 결과를 나타낸 것이다.Figures 2a to 2e show the results of testing the anticancer effect of the N-acylhydrazone derivative in a human cervical cancer cell (HeLa CCL2) transplantation model.
도 3a 내지 3e은 N-아실히드라존 유도체의 혈장 내 안정성 시험 결과이다.3A to 3E are results of a stability test in plasma of an N-acylhydrazone derivative.
도 4a 내지 4e는 N-아실히드라존 유도체의 간 미세소관 내 대사안정성 시험 결과이다.4A to 4E are results of a metabolic stability test in liver microtubules of N-acylhydrazone derivatives.
도 5a 내지 5c는 N-아실히드라존 유도체의 약물동태(PK) 시험 결과이다.5a to 5c are pharmacokinetic (PK) test results of N-acylhydrazone derivatives.
도 6a 및 6b는 N-아실히드라존 유도체의 상처회복시험(wound healing assay) 결과이다.6A and 6B are results of a wound healing assay of N-acylhydrazone derivatives.
도 7a 및 7b는 N-아실히드라존 유도체의 침투시험(in vitro invasion assay) 결과이다.7a and 7b are results of an in vitro invasion assay of N-acylhydrazone derivatives.
도 8은 N-아실히드라존 유도체의 시간에 따른 각 장기별 타겟팅 변화를 나타낸 것이다.8 shows the targeting change for each organ according to time of the N-acylhydrazone derivative.
도 9는 N-아실히드라존 유도체-형광체 접합체의 세포내 투과를 관찰한 결과이다.9 is a result of observing the intracellular permeation of the N-acylhydrazone derivative-fluorescent conjugate.
도 10은 N-아실히드라존 유도체-형광체 접합체의 암조직에 대한 타겟팅을 측정한 결과이다.10 is a result of measuring the targeting of the N-acylhydrazone derivative-phosphor conjugate to cancer tissue.
도 11 및 12는 N-아실히드라존 유도체-형광체 접합체의 각 조직별 타겟팅을 측정한 결과이다.11 and 12 show the results of measuring the targeting of the N-acylhydrazone derivative-phosphor conjugate to each tissue.
도 13은 N-아실히드라존 유도체-형광체 접합체의 암조직과 근육조직에 대한 타겟팅을 측정한 결과이다.13 is a result of measuring the targeting of the N-acylhydrazone derivative-fluorescent conjugate to cancer tissue and muscle tissue.
도 14은 N-아실히드라존 유도체-리간드 접합체의 암조직 축적량을 방사성 동위원소를 이용하여 관찰한 결과이다.14 is a result of observing the amount of cancer tissue accumulation of an N-acylhydrazone derivative-ligand conjugate using a radioactive isotope.
도 15는 N-아실히드라존 유도체-리간드 접합체의 시간에 따른 암조직 타겟팅을 측정한 결과이다.15 is a result of measuring cancer tissue targeting of the N-acylhydrazone derivative-ligand conjugate over time.
도 16은 N-아실히드라존 유도체-리간드 접합체의 암세포에 대한 선택적 투과율을 측정한 결과이다.16 is a result of measuring the selective permeability of an N-acylhydrazone derivative-ligand conjugate to cancer cells.
도 17은 N-아실히드라존 유도체-리간드 접합체 및 다른 N-아실히드라존 유도체의 암세포에 대한 경쟁적 투과율을 측정한 결과이다.17 shows the results of measuring the competitive permeability of N-acylhydrazone derivative-ligand conjugates and other N-acylhydrazone derivatives against cancer cells.
본 명세서에서, 용어 "할로겐"은 다른 언급이 없으면 F, Cl, Br 또는 I를 의미한다.As used herein, the term "halogen" means F, Cl, Br or I, unless otherwise stated.
용어 "알킬"은, 달리 명시되지 않는 한, 선형 또는 분지형의 포화된 탄화수소 잔기를 의미한다. 예를 들어, "C1-6알킬"은 1 내지 6개 탄소로 골격이 이루어진 알킬을 의미한다. 구체적으로 C1-6알킬은 메틸, 에틸, n-프로필, i-프로필, n-부틸, i-부틸, t-부틸, n-펜틸, i-펜틸, t-펜틸, sec-펜틸, 네오펜틸, 헥실 등을 포함할 수 있다.The term “alkyl,” unless otherwise specified, refers to a linear or branched saturated hydrocarbon moiety. For example, "C 1-6 alkyl" refers to alkyl having a backbone of 1 to 6 carbons. Specifically, C 1-6 alkyl is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, sec-pentyl, neopentyl , hexyl, and the like.
용어 "알콕시"는, 달리 명시되지 않는 한, 선형 또는 분지형의 알킬-옥시 잔기를 의미한다. 예를 들어, "C1-6알콕시"는 1 내지 6개 탄소로 골격이 이루어진 알킬-옥시를 의미한다. 구체적으로 C1-6알콕시는 메톡시, 에톡시, n-프로폭시, i-프로폭시, n-부톡시, i-부톡시, t-부톡시, n-펜톡시, i-펜톡시, t-펜톡시, sec-펜톡시, 네오펜톡시, 헥실옥시 등을 포함할 수 있다.The term "alkoxy", unless otherwise specified, refers to a linear or branched alkyl-oxy moiety. For example, "C 1-6 alkoxy" means alkyl-oxy with a backbone of 1 to 6 carbons. Specifically, C 1-6 alkoxy is methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, t-butoxy, n-pentoxy, i-pentoxy, t -pentoxy, sec-pentoxy, neopentoxy, hexyloxy, and the like may be included.
용어 "할로알킬" 또는 "할로알콕시"는 하나 이상의 할로겐으로 치환된 알킬 또는 알콕시를 의미한다. 구체적으로, 할로알킬 또는 할로알콕시는 동종 또는 이종의 할로겐이 1개 이상 치환된 알킬 또는 알콕시일 수 있다.The term “haloalkyl” or “haloalkoxy” means an alkyl or alkoxy substituted with one or more halogens. Specifically, haloalkyl or haloalkoxy may be alkyl or alkoxy in which one or more homogeneous or heterogeneous halogens are substituted.
용어 "치환"은, 지정된 원자 상의 원자가(valence)를 초과하지 않으면서 이러한 치환으로부터 화학적으로 안정한 화합물이 되도록, 분자 구조체 내의 수소 원자를 치환기로 대체하는 것을 지칭한다. 예를 들어 "그룹 A가 치환기 B로 치환"된다는 것은, 그룹 A의 골격을 구성하는 탄소 등의 원자에 결합된 수소 원자가 치환기 B로 대체되어, 그룹 A와 치환기 B가 공유 결합을 형성함을 의미할 수 있다.The term “substitution” refers to the replacement of a hydrogen atom in a molecular structure with a substituent such that the compound is chemically stable from such substitution without exceeding the valence on the designated atom. For example, "group A is substituted with substituent B" means that a hydrogen atom bonded to an atom such as carbon constituting the backbone of group A is replaced with substituent B, so that group A and substituent B form a covalent bond can do.
본 발명에 따르면, 암에 대한 타겟팅이 뛰어난 새로운 N-아실히드라존 유도체-표지자 접합체, 및 상기 접합체에 도입되는 새로운 구조의 N-아실히드라존 유도체가 제공된다.According to the present invention, a novel N-acylhydrazone derivative-marker conjugate excellent in targeting to cancer, and an N-acylhydrazone derivative having a novel structure introduced into the conjugate are provided.
N-아실히드라존 유도체N-acylhydrazone derivatives
상기 N-아실히드라존 유도체는 하기 화학식 1의 일반식 구조를 갖는다.The N-acylhydrazone derivative has the general structure of Formula 1 below.
[화학식 1][Formula 1]
Figure PCTKR2021007223-appb-img-000006
Figure PCTKR2021007223-appb-img-000006
상기 화학식 1에서, R은
Figure PCTKR2021007223-appb-img-000007
,
Figure PCTKR2021007223-appb-img-000008
,
Figure PCTKR2021007223-appb-img-000009
또는
Figure PCTKR2021007223-appb-img-000010
이고; A는 -O- 또는 -S-이고; R1은 H, C1-6알킬, C1-6알콕시카보닐C1-3알킬, 또는 C1-6알콕시C1-3알킬이고; R2는 할로겐, C1-6알킬 또는 할로C1-6알킬이고; R3은 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 할로C1-6알콕시이고; R4 및 R5는 각각 독립적으로 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 C1-6알킬카보닐아미노이다.In Formula 1, R is
Figure PCTKR2021007223-appb-img-000007
,
Figure PCTKR2021007223-appb-img-000008
,
Figure PCTKR2021007223-appb-img-000009
or
Figure PCTKR2021007223-appb-img-000010
ego; A is -O- or -S-; R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl; R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl; R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy; R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino.
상기 C1-6알킬은 C1-3알킬, C3-6알킬, C2-4알킬, C2-6알킬 등을 포함할 수 있다. 또한, 상기 C1-6알콕시는 C1-3알콕시, C3-6알콕시, C2-4알콕시, C2-6알콕시 등을 포함할 수 있다. 또한, 상기 할로C1-6알킬 및 할로C1-6알콕시는 각각 1개 내지 10개, 또는 1개 내지 3개의 동일하거나 서로 다른 할로겐을 가질 수 있다.The C 1-6 alkyl may include C 1-3 alkyl, C 3-6 alkyl, C 2-4 alkyl, C 2-6 alkyl, and the like. In addition, the C 1-6 alkoxy may include C 1-3 alkoxy, C 3-6 alkoxy, C 2-4 alkoxy, C 2-6 alkoxy, and the like. In addition, the haloC 1-6 alkyl and haloC 1-6 alkoxy may each have 1 to 10, or 1 to 3, identical or different halogens.
일 구현예에 따르면, 상기 화학식 1에서, R은
Figure PCTKR2021007223-appb-img-000011
일 수 있다.According to one embodiment, in Formula 1, R is
Figure PCTKR2021007223-appb-img-000011
can be
다른 구현예에 따르면, 상기 화학식 1에서, R은
Figure PCTKR2021007223-appb-img-000012
,
Figure PCTKR2021007223-appb-img-000013
,
Figure PCTKR2021007223-appb-img-000014
또는
Figure PCTKR2021007223-appb-img-000015
일 수 있다.According to another embodiment, in Formula 1, R is
Figure PCTKR2021007223-appb-img-000012
,
Figure PCTKR2021007223-appb-img-000013
,
Figure PCTKR2021007223-appb-img-000014
or
Figure PCTKR2021007223-appb-img-000015
can be
또 다른 구현예에 따르면, 상기 화학식 1에서, R1은 H, -CH3, -CH2CO2CH3, -CH2CO2CH2CH3, -CH2CH2OCH2CH3 또는 -CH2CH2OCH3이다.According to another embodiment, in Formula 1, R 1 is H, -CH 3 , -CH 2 CO 2 CH 3 , -CH 2 CO 2 CH 2 CH 3 , -CH 2 CH 2 OCH 2 CH 3 or - CH 2 CH 2 OCH 3 .
또 다른 구현예에 따르면, 상기 화학식 1에서, R2 및 R3은 각각 독립적으로 H, F, Cl, Br, -CH3 -CF3, -OCH3 또는 -OCF3이다.According to another embodiment, in Formula 1, R 2 and R 3 are each independently H, F, Cl, Br, -CH 3 -CF 3, -OCH 3 or -OCF 3.
또 다른 구현예에 따르면, 상기 화학식 1에서, R4 및 R5는 각각 독립적으로 H, Cl, -CH3, -OCH3, 또는 -NHCOCH3이다.According to another embodiment, in Formula 1, R 4 and R 5 are each independently H, Cl, -CH 3, -OCH 3, or -NHCOCH 3.
또 다른 구현예에 따르면, 상기 화학식 1에서, R1은 H, -CH3, -CH2CO2CH3, -CH2CO2CH2CH3, -CH2CH2OCH2CH3 또는 -CH2CH2OCH3이고; R2는 Cl, Br, -CH3 또는 -CF3이고; R3은 H, F, Cl, -CH3, -OCH3, 또는 -OCF3이고; R4 및 R5는 각각 독립적으로 H, Cl, -CH3, -OCH3, 또는 -NHCOCH3이다.According to another embodiment, in Formula 1, R 1 is H, -CH 3 , -CH 2 CO 2 CH 3 , -CH 2 CO 2 CH 2 CH 3 , -CH 2 CH 2 OCH 2 CH 3 or - CH 2 CH 2 OCH 3 ; R 2 is Cl, Br, —CH 3 or —CF 3 ; R 3 is H, F, Cl, -CH 3 , -OCH 3, or -OCF 3, and; R 4 and R 5 are each independently H, Cl, -CH 3, -OCH 3, or -NHCOCH 3.
또 다른 구현예에 따르면, 상기 화학식 1에서, R1은 -CH2CO2CH3, -CH2CO2CH2CH3 또는 -CH2CH2OCH2CH3이고; R2는 Cl, Br 또는 -CH3이고; R3은 H 또는 -OCH3이고; R4 및 R5는 각각 독립적으로 H, Cl, -CH3 또는 -OCH3이다.According to another embodiment, in Formula 1, R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ; R 2 is Cl, Br or —CH 3 ; R 3 is H or —OCH 3 ; R 4 and R 5 are each independently H, Cl, -CH 3 or -OCH 3 .
또 다른 구현예에 따르면, 상기 화학식 1에서, R은
Figure PCTKR2021007223-appb-img-000016
,
Figure PCTKR2021007223-appb-img-000017
,
Figure PCTKR2021007223-appb-img-000018
또는
Figure PCTKR2021007223-appb-img-000019
이고; R1은 -CH2CO2CH3, -CH2CO2CH2CH3 또는 -CH2CH2OCH2CH3이고; R2는 Cl 또는 -CH3이고; R3은 H 또는 -OCH3일 수 있다.According to another embodiment, in Formula 1, R is
Figure PCTKR2021007223-appb-img-000016
,
Figure PCTKR2021007223-appb-img-000017
,
Figure PCTKR2021007223-appb-img-000018
or
Figure PCTKR2021007223-appb-img-000019
ego; R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ; R 2 is Cl or —CH 3 ; R 3 may be H or —OCH 3 .
상기 화학식 1의 N-아실히드라존 유도체의 구체적인 예시들은 아래 화학식 1a 내지 1e의 구조를 포함할 수 있다.Specific examples of the N-acylhydrazone derivative of Formula 1 may include structures of Formulas 1a to 1e below.
[화학식 1a][Formula 1a]
Figure PCTKR2021007223-appb-img-000020
Figure PCTKR2021007223-appb-img-000020
[화학식 1b][Formula 1b]
Figure PCTKR2021007223-appb-img-000021
Figure PCTKR2021007223-appb-img-000021
[화학식 1c][Formula 1c]
Figure PCTKR2021007223-appb-img-000022
Figure PCTKR2021007223-appb-img-000022
[화학식 1d][Formula 1d]
Figure PCTKR2021007223-appb-img-000023
Figure PCTKR2021007223-appb-img-000023
[화학식 1e][Formula 1e]
Figure PCTKR2021007223-appb-img-000024
Figure PCTKR2021007223-appb-img-000024
상기 식에서 R1, R2, R3, R4 및 R5는 앞서의 화학식 1 또는 이의 구현예에서 예시한 바와 같다.In the above formula, R 1 , R 2 , R 3 , R 4 and R 5 are as exemplified in Formula 1 or an embodiment thereof.
상기 화학식 1a의 일 구현예에서, R1은 H, C1-6알킬, C1-6알콕시카보닐C1-3알킬, 또는 C1-6알콕시C1-3알킬이고; R2는 할로겐, C1-6알킬 또는 할로C1-6알킬이고; R3은 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 할로C1-6알콕시이고; R4 및 R5는 각각 독립적으로 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 C1-6알킬카보닐아미노이다.In one embodiment of Formula 1a, R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl; R 2 is halogen, C 1-6 alkyl or haloC 1-6 alkyl; R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy; R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino.
상기 화학식 1b의 일 구현예에서, R1은 C1-3알콕시카보닐C1-3알킬이고; R2는 할로겐이고; R3은 C1-3알콕시이다.In one embodiment of Formula 1b, R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl; R 2 is halogen; R 3 is C 1-3 alkoxy.
상기 화학식 1c의 일 구현예에서, R1은 C1-3알콕시카보닐C1-3알킬이고; R2는 할로겐이고; R3은 H 또는 C1-3알콕시이다.In one embodiment of Formula 1c, R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl; R 2 is halogen; R 3 is H or C 1-3 alkoxy.
상기 화학식 1d의 일 구현예에서, R1은 C1-3알콕시카보닐C1-3알킬이다.In one embodiment of Formula 1d, R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl.
상기 화학식 1e의 일 구현예에서, R1은 C1-3알콕시카보닐C1-3알킬이고; R2는 할로겐 또는 C1-3알킬이다.In one embodiment of Formula 1e, R 1 is C 1-3 alkoxycarbonylC 1-3 alkyl; R 2 is halogen or C 1-3 alkyl.
구체적인 일례로서, 상기 화학식 1의 N-아실히드라존 유도체는 하기로 이루어진 군에서 선택될 수 있다:As a specific example, the N-acylhydrazone derivative of Formula 1 may be selected from the group consisting of:
1. (E)-N'-[(2-클로로-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;1. (E)—N′-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
2. (E)-N'-[(2-클로로-1-메틸-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;2. (E)—N′-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
3. 에틸 (E)-2-{2-클로로-3-[(2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸]-1H-인돌-1-일}아세테이트;3. ethyl (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl]-1H-indol-1-yl}acetate;
4. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;4. (E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
5. (E)-N'-{[2-브로모-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;5. (E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
6. (E)-N'-{[1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;6. (E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbo hydrazide;
7. (E)-N'-{[2-클로로-1-(2-메톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;7. (E)—N′-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
8. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;8. (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbo hydrazide;
9. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;9. (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbo hydrazide;
10. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;10. (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbo hydrazide;
11. (E)-N'-{[2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;11. (E)—N′-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
12. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;12. (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran -2-carbohydrazide;
13. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;13. (E)-N'-{[2-Chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydra Gide;
14. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;14. (E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydrazide;
15. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;15. (E)-N'-{[2-Chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2- carbohydrazide;
16. (E)-5-클로로-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}벤조퓨란-2-카보히드라지드;16. (E)-5-chloro-N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbohydrazide;
17. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,7-디메틸벤조퓨란-2-카보히드라지드;17. (E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbohydrazide;
18. (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,6-디메톡시벤조퓨란-2-카보히드라지드; 18. (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbohydrazide ;
19. (E)-N'-{2-[2-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌]히드라진-1-카보닐}벤조퓨란-5-일)아세트아미드;19. (E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl}benzofuran -5-yl)acetamide;
20. (E)-에틸-2-(3-((2-(4,6-디메톡시벤조퓨란-2-카보닐)히드라진일리덴)메틸)-2-메틸-1H-인돌-1-일)아세테이트;20. (E)-ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazinylidene)methyl)-2-methyl-1H-indol-1-yl )acetate;
21. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드;21. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2-carbo hydrazide;
22. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드;22. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2-carbohydra Gide;
23. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;23. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbo hydrazide;
24. (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;24. (E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide;
25. (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드;25. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbohydrazide ;
26. (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트; 및26. (E)-Ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indole-1- work) acetate; and
27. (E)-메틸 2-(2-메틸-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트.27. (E)-Methyl 2-(2-methyl-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indole-1- 1) Acetate.
구체적인 다른 예로서, 상기 화학식 1의 N-아실히드라존 유도체는 하기로 이루어진 군에서 선택될 수 있다:As another specific example, the N-acylhydrazone derivative of Formula 1 may be selected from the group consisting of:
21. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드;21. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2-carbo hydrazide;
22. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드;22. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2-carbohydra Gide;
23. (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;23. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbo hydrazide;
24. (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;24. (E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide;
25. (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드; 및25. (E)-N'-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbohydrazide ; and
26. (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트.26. (E)-Ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indole-1- 1) Acetate.
약물-표지자 접합체drug-marker conjugate
상기 화학식 1의 N-아실히드라존 유도체는 세포증식성 질환의 진단 및 효과적인 치료를 위해 표지자와 접합될 수 있다.The N-acylhydrazone derivative of Formula 1 may be conjugated to a marker for diagnosis and effective treatment of cell proliferative diseases.
상기 표지자는 방사성 동위원소의 리간드, 또는 형광체일 수 있다.The marker may be a ligand of a radioisotope or a fluorescent substance.
상기 방사성 동위원소의 리간드는 디페록사민(DFO), 디에틸렌트리아민펜타아세트산(DTPA) 및 1,4,7,10-테트라아자시클로도데칸-1,4,7,10-테트라아세트산(DOTA)로 이루어진 군에서 선택되는 1종 이상일 수 있으나, 이에 한정되지 않는다. 상기 방사성 동위원소는 89Zr, 60Cu, 64Cu, 223Ra, 131I, 89Sr, 32P 및 188Re로 이루어진 군에서 선택되는 1종 이상일 수 있다.Ligands of the radioactive isotopes include deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). ) may be one or more selected from the group consisting of, but is not limited thereto. The radioactive isotope may be at least one selected from the group consisting of 89 Zr, 60 Cu, 64 Cu, 223 Ra, 131 I, 89 Sr, 32 P, and 188 Re.
상기 형광체는 시아닌 3, 시아닌 5, 시아닌 5.5 및 시아닌 7로 이루어진 군에서 선택되는 1종 이상일 수 있으나, 이에 한정되지 않는다. The phosphor may be at least one selected from the group consisting of cyanine 3, cyanine 5, cyanine 5.5, and cyanine 7, but is not limited thereto.
그러나 상기 리간드 및 형광체는 이에 한정되지 않고, 질병의 진단 및 효과적인 치료를 위해 사용되는 다른 리간드 또는 형광체도 가능하다.However, the ligand and the fluorescent substance are not limited thereto, and other ligands or fluorescent substances used for diagnosis and effective treatment of diseases are possible.
상기 N-아실히드라존 유도체는 말단에서 상기 표지자와 접합된다. 예를 들어 상기 화학식 1의 R1의 말단에서 상기 표지자와 접합될 수 있다. The N-acylhydrazone derivative is terminally conjugated with the marker. For example, it may be conjugated to the marker at the end of R 1 in Formula 1 above.
상기 N-아실히드라존 유도체와 상기 표지자의 접합은 약물과 표지자의 일반적인 접합 방식을 채용할 수 있으며, 예를 들어 아미드 결합(-NH-C(=O)-) 또는 티오아미드 결합(-NH-C(=S)-)에 의해 접합될 수 있다.The conjugation of the N-acylhydrazone derivative and the marker may employ a general conjugation method between the drug and the marker, for example, an amide bond (-NH-C(=O)-) or a thioamide bond (-NH- C(=S)-).
이를 위해, 상기 접합체는 말단이 아민화된 N-아실히드라존 유도체를 포함할 수 있다. 구체적으로, 상기 N-아실히드라존 유도체의 R1의 말단이 아민화되어 상기 표지자와 접합될 수 있다.To this end, the conjugate may include a terminally aminated N-acylhydrazone derivative. Specifically, the terminal of R 1 of the N-acylhydrazone derivative may be aminated to be conjugated to the marker.
또한, 상기 N-아실히드라존 유도체는 링커에 의해 상기 표지자와 접합될 수 있다. 상기 링커는 p-NCS-벤젠을 포함할 수 있다. 예를 들어, -C(=S)-NH-페닐렌-NH-C(=S)-일 수 있다. 또한, -C(=O)- 일 수 있다. 그러나 상기 링커는 이에 한정되지 않고, 그 외 접합에 사용되는 다른 링커도 가능하다.In addition, the N-acylhydrazone derivative may be conjugated to the marker by a linker. The linker may include p-NCS-benzene. For example, it may be -C(=S)-NH-phenylene-NH-C(=S)-. Also, it may be -C(=O)-. However, the linker is not limited thereto, and other linkers used for conjugation are also possible.
일 구현예에 따르면, 상기 N-아실히드라존 유도체와 표지자의 접합체는 하기 화학식 2로 표시될 수 있다:According to one embodiment, the conjugate of the N-acylhydrazone derivative and the marker may be represented by the following formula (2):
[화학식 2][Formula 2]
Figure PCTKR2021007223-appb-img-000025
Figure PCTKR2021007223-appb-img-000025
상기 식에서 R, R2, R3은 상기 화학식 1에서 정의된 바와 같고; R0은 상기 화학식 1의 R1에서 하나의 H가 제거된 그룹이고; L은 단일 결합, 또는 링커로부터 유래된 그룹이고; A는 표지자로부터 유래된 그룹이다.In the above formula, R, R 2 , R 3 are as defined in Formula 1 above; R 0 is a group in which one H is removed from R 1 in Formula 1; L is a single bond or a group derived from a linker; A is a group derived from a marker.
상기 화학식 2에서 R0은 상기 화학식 1의 R1에서 유래된 그룹이다. 즉 R0은 상기 화학식 1의 R1(즉 1가 라디칼 그룹)에서 하나의 H가 제거되어 추가적인 라디칼이 형성된 그룹(즉 2가 라디칼 그룹)이다. 구체적으로 R0은 -(CH2)m-, -(CH2)n-COO-(CH2)m-, 또는 -(CH2)n-O-(CH2)m-일 수 있고, 여기서 m은 1 내지 6의 정수이고, n은 1 내지 3의 정수이다. 보다 구체적으로, -CH2, -CH2CO2CH2, -CH2CO2CH2CH2, -CH2CH2OCH2CH2 또는 -CH2CH2OCH2이다 In Formula 2, R 0 is a group derived from R 1 in Formula 1 above. That is, R 0 is a group (ie, a divalent radical group) in which an additional radical is formed by removing one H from R 1 (ie, a monovalent radical group) of Formula 1 above. Specifically, R 0 may be -(CH 2 ) m -, -(CH 2 ) n -COO-(CH 2 ) m -, or -(CH 2 ) n -O-(CH 2 ) m -, wherein m is an integer from 1 to 6, and n is an integer from 1 to 3. More specifically, -CH 2 , -CH 2 CO 2 CH 2 , -CH 2 CO 2 CH 2 CH 2 , -CH 2 CH 2 OCH 2 CH 2 or -CH 2 CH 2 OCH 2
상기 N-아실히드라존 유도체와 표지자가 별도의 링커 없이 직접 접합되는 경우, 상기 화학식 2에서 L은 단일 결합일 수 있다. 즉, L의 단일 결합은 링커가 없이 직접 결합되는 것을 의미할 수 있다. When the N-acylhydrazone derivative and the label are directly conjugated without a separate linker, L in Formula 2 may be a single bond. That is, the single bond of L may mean that it is directly bonded without a linker.
또는 상기 N-아실히드라존 유도체와 표지자가 링커를 매개로 접합되는 경우에는, 상기 화학식 2에서 L은 링커로부터 유래된 그룹일 수 있다. 상기 링커의 종류는 앞서 예시한 바와 같으며, 예를 들어 p-NCS-벤젠을 매개로 접합되는 경우에는, 상기 화학식 2에서 L은 -C(=S)-NH-페닐렌-NH-C(=S)- 또는 -C(=O)-을 포함할 수 있다.Alternatively, when the N-acylhydrazone derivative and the marker are conjugated via a linker, L in Formula 2 may be a group derived from a linker. The type of the linker is as exemplified above, and, for example, when conjugated via p-NCS-benzene, L in Formula 2 is -C(=S)-NH-phenylene-NH-C( =S)- or -C(=O)-.
구체적인 예로서, 상기 화학식 2에서 R, R2, R3은 상기 화학식 1에서 정의된 바와 같고; R0은 상기 화학식 1의 R1에서 하나의 H가 제거된 그룹이고; L은 단일 결합, 또는 링커로부터 유래된 그룹이고; A는 방사성 동위원소의 리간드 또는 형광체로부터 유래된 그룹이다.As a specific example, in Formula 2, R, R 2 , and R 3 are as defined in Formula 1; R 0 is a group in which one H is removed from R 1 in Formula 1; L is a single bond or a group derived from a linker; A is a group derived from a ligand or fluorescent substance of a radioisotope.
상기 화학식 2에서 A는 방사성 동위원소의 리간드 또는 형광체로부터 유래된 그룹일 수 있다.In Formula 2, A may be a group derived from a ligand or a phosphor of a radioisotope.
구체적으로, 상기 화학식 2에서 A는 방사성 동위원소의 리간드로부터 유래된 그룹으로서, 상기 리간드에서 하나의 H 또는 말단기가 제거되어 라디칼이 형성된 그룹일 수 있다. Specifically, in Formula 2, A is a group derived from a ligand of a radioisotope, and may be a group in which one H or a terminal group is removed from the ligand to form a radical.
예를 들어 A가 디페록사민(DFO)으로부터 유래된 그룹일 경우, 디페록사민에서 하나의 H가 제거되어 A는 아래와 같은 구조를 가질 수 있다.For example, when A is a group derived from deferoxamine (DFO), one H is removed from deferoxamine so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000026
Figure PCTKR2021007223-appb-img-000026
A가 디에틸렌트리아민펜타아세트산(DTPA)으로부터 유래된 그룹일 경우, 디에틸렌트리아민펜타아세트산에서 하나의 OH가 제거되어 A는 아래와 같은 구조를 가질 수 있다.When A is a group derived from diethylenetriaminepentaacetic acid (DTPA), one OH is removed from diethylenetriaminepentaacetic acid, so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000027
Figure PCTKR2021007223-appb-img-000027
A가 1,4,7,10-테트라아자시클로도데칸-1,4,7,10-테트라아세트산(DOTA)으로부터 유래된 그룹일 경우, 1,4,7,10-테트라아자시클로도데칸-1,4,7,10-테트라아세트산에서 하나의 OH가 제거되어 A는 아래와 같은 구조를 가질 수 있다.When A is a group derived from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7,10-tetraazacyclododecane- One OH is removed from 1,4,7,10-tetraacetic acid, so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000028
Figure PCTKR2021007223-appb-img-000028
또한, 상기 화학식 2에서 A는 형광체로부터 유래된 그룹으로서, 상기 형광체에서 하나의 H 또는 말단기가 제거되어 라디칼이 형성된 그룹일 수 있다. In addition, in Formula 2, A is a group derived from a phosphor, and may be a group in which one H or a terminal group is removed from the phosphor to form a radical.
예를 들어, A가 시아닌 3(Cy3)으로부터 유래된 그룹일 경우, 시아닌 3에서 말단기가 제거되어 A는 아래와 같은 구조를 가질 수 있다.For example, when A is a group derived from cyanine 3 (Cy3), a terminal group is removed from cyanine 3, so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000029
Figure PCTKR2021007223-appb-img-000029
A가 시아닌 5(Cy5)으로부터 유래된 그룹일 경우, 시아닌 5에서 말단기가 제거되어 A는 아래와 같은 구조를 가질 수 있다.When A is a group derived from cyanine 5 (Cy5), the terminal group is removed from cyanine 5 so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000030
Figure PCTKR2021007223-appb-img-000030
A가 시아닌 5.5(Cy5.5)으로부터 유래된 그룹일 경우, 시아닌 5.5에서 말단기가 제거되어 A는 아래와 같은 구조를 가질 수 있다.When A is a group derived from cyanine 5.5 (Cy5.5), the terminal group is removed from cyanine 5.5, so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000031
Figure PCTKR2021007223-appb-img-000031
A가 시아닌 7(Cy7)으로부터 유래된 그룹일 경우, 시아닌 7에서 말단기가 제거되어 A는 아래와 같은 구조를 가질 수 있다.When A is a group derived from cyanine 7 (Cy7), a terminal group is removed from cyanine 7, so that A may have the following structure.
Figure PCTKR2021007223-appb-img-000032
Figure PCTKR2021007223-appb-img-000032
즉, N-아실히드라존 유도체와 표지자의 접합체와 관련하여 상기 화학식 2에서 A는 위 나열된 그룹으로부터 선택되는 어느 하나일 수 있다. That is, in relation to the conjugate of the N-acylhydrazone derivative and the marker, in Formula 2, A may be any one selected from the groups listed above.
N-아실히드라존 유도체의 경우 앞서 화학식 1에 정의된 N-아실히드라존 유도체가 화학식 2에 쉽게 적용될 수 있고, 이는 하기 실시예들을 참고하여 통상의 기술자가 용이하게 화합물을 제조할 수 있다.In the case of the N-acylhydrazone derivative, the N-acylhydrazone derivative defined in Formula 1 above can be easily applied to Formula 2, and a person skilled in the art can easily prepare a compound with reference to the following Examples.
N-아실히드라존 유도체와 표지자의 접합체와 관련하여 구체적인 일례로서 아래 화학식 2a 내지 2g를 기재하며, N-아실히드라존 유도체는 아래 일예시의 화합물을 대체하여 쉽게 치환 가능한 바 본 발명은 아래 실시양태로만 국한되는 것은 아니다. As a specific example with respect to the conjugate of an N-acylhydrazone derivative and a marker, the following formulas 2a to 2g are described, and the N-acylhydrazone derivative can be easily substituted by replacing the compounds of the examples below. is not limited to
하기 화학식 2a로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.A conjugate represented by the following formula (2a), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2a][Formula 2a]
Figure PCTKR2021007223-appb-img-000033
Figure PCTKR2021007223-appb-img-000033
본 발명의 일실시양태에 따르면, 링커로 p-NCS-벤젠을 매개하여 접합함으로서 상기 화학식 2a로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커 없이 결합한 형태를 제공할 수도 있다. According to an embodiment of the present invention, the conjugate represented by Formula 2a, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof can be provided by conjugation via p-NCS-benzene with a linker. If necessary, a combined form without a linker may be provided.
구체적인 다른 예로서, 하기 화학식 2b로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following Chemical Formula 2b, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2b][Formula 2b]
Figure PCTKR2021007223-appb-img-000034
Figure PCTKR2021007223-appb-img-000034
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2b로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다. According to an embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2b, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
구체적인 다른 예로서, 하기 화학식 2c로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following Chemical Formula 2c, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2c][Formula 2c]
Figure PCTKR2021007223-appb-img-000035
Figure PCTKR2021007223-appb-img-000035
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2c로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다.According to one embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2c, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
구체적인 다른 예로서, 하기 화학식 2d로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following Chemical Formula 2d, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2d][Formula 2d]
Figure PCTKR2021007223-appb-img-000036
Figure PCTKR2021007223-appb-img-000036
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2d로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다.According to one embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2d, a stereoisomer or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
구체적인 다른 예로서, 하기 화학식 2e로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following Chemical Formula 2e, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2e][Formula 2e]
Figure PCTKR2021007223-appb-img-000037
Figure PCTKR2021007223-appb-img-000037
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2e로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다.According to one embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2e, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
구체적인 다른 예로서, 하기 화학식 2f로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following formula (2f), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2f][Formula 2f]
Figure PCTKR2021007223-appb-img-000038
Figure PCTKR2021007223-appb-img-000038
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2f로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다.According to one embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2f, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
구체적인 다른 예로서, 하기 화학식 2g로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염이 제공된다.As another specific example, a conjugate represented by the following Chemical Formula 2g, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 2g][Formula 2g]
Figure PCTKR2021007223-appb-img-000039
Figure PCTKR2021007223-appb-img-000039
본 발명의 일실시양태에 따르면, 링커 없이 직접 접합하여 상기 화학식 2g로 표시되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 제공할 수 있다. 필요에 따라 링커로 매개된 구조로 제공할 수도 있다.According to an embodiment of the present invention, it is possible to provide a conjugate represented by Formula 2g, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof by direct conjugation without a linker. If necessary, it may be provided in a structure mediated by a linker.
상기 N-아실히드라존 유도체와 표지자의 접합체를 제조하는 방법은 특별히 한정되지 않으며, 각각의 구조에 적합한 방법으로 수행될 수 있다. 예를 들어 상기 화학식 1에서 말단이 아민화된 N-아실히드라존 유도체를 얻은 뒤 이를 표지자와 반응시켜 접합체를 제조할 수 있다. A method for preparing the conjugate of the N-acylhydrazone derivative and the marker is not particularly limited and may be carried out by a method suitable for each structure. For example, a conjugate may be prepared by obtaining an N-acylhydrazone derivative having an end amination in Chemical Formula 1 and reacting it with a marker.
일 구현예에 따른 접합체의 제조하는 방법은 (1) 상기 화학식 1에서 말단이 아민화된 N-아실히드라존 유도체 또는 이의 염을 제조하는 단계; 및 (2) 상기 아민화된 N-아실히드라존 유도체 또는 이의 염을 표지자와 접합하는 단계를 포함할 수 있다.A method for preparing a conjugate according to an embodiment comprises the steps of (1) preparing an N-acylhydrazone derivative or a salt thereof having an terminal aminated in Formula 1; and (2) conjugating the aminated N-acylhydrazone derivative or salt thereof with a marker.
단계 (1)에서, 말단이 아민화된 N-아실히드라존 유도체 또는 이의 염을 제조하기 위해, 상기 화학식 1의 N-아실히드라존 유도체의 합성 절차 이후 말단 그룹을 아민화할 수 있다. 또는 상기 화학식 1에서 아민화시킬 치환기를 미리 선정하고, 상기 치환기를 도입하기 위한 반응물질의 말단을 미리 아민화한 뒤, 상기 화학식 1의 N-아실히드라존 유도체의 합성 반응에 투입할 수 있다.In step (1), in order to prepare the terminally aminated N-acylhydrazone derivative or a salt thereof, the terminal group may be aminated after the synthesis procedure of the N-acylhydrazone derivative of Formula 1 above. Alternatively, a substituent to be aminated in Formula 1 may be selected in advance, and a terminal of a reactant for introducing the substituent may be aminationed in advance, and then added to the synthesis reaction of the N-acylhydrazone derivative of Formula 1 above.
Figure PCTKR2021007223-appb-img-000040
Figure PCTKR2021007223-appb-img-000040
상기 식에서, R, R2, R3은 상기 화학식 1에서 정의된 바와 같고; R0은 상기 화학식 1의 R1에서 하나의 H가 제거된 그룹이고; P는 보호기이고; X는 할로겐이다.In the above formula, R, R 2 , R 3 are as defined in Formula 1 above; R 0 is a group in which one H is removed from R 1 in Formula 1; P is a protecting group; X is halogen.
예를 들어 상기 반응식에서 보듯이 (i) 상기 화학식 1의 N-아실히드라존 유도체의 합성 절차 이후 R1을 아민-보호된 할로겐화아민 화합물과 반응시키거나, 또는 (ii) R1이 아민화 및 보호된 인돌 화합물을 미리 제조한 뒤 이를 히드라존 화합물과 반응시키고 이후 보호기를 제거하여, R1의 말단이 아민화된 N-아실히드라존 유도체를 얻을 수 있다.For example, as shown in the reaction scheme, (i) after the synthesis of the N-acylhydrazone derivative of Formula 1, R 1 is reacted with an amine-protected halogenated amine compound, or (ii) R 1 is amination and After preparing a protected indole compound in advance, it is reacted with a hydrazone compound and then the protecting group is removed to obtain an N-acylhydrazone derivative in which the terminal of R 1 is aminated.
또한 상기 아민화된 N-아실히드라존 유도체는 염산 등과 더 반응시켜 염으로 형성한 뒤 다음 반응에 사용함으로써 최종 수율을 높일 수 있다. In addition, the aminated N-acylhydrazone derivative may be further reacted with hydrochloric acid to form a salt, and then used in the next reaction to increase the final yield.
단계 (2)에서, 아민화된 N-아실히드라존 유도체 또는 이의 염과 표지자의 접합은 이 분야에 사용되는 통상적인 방식으로 수행될 수 있다. 예를 들어 상기 접합은 아미드화 반응 또는 티오아미드화 반응을 통해 수행될 수 있으나, 이에 한정되지 않는다.In step (2), the conjugation of the aminated N-acylhydrazone derivative or salt thereof with the marker can be carried out in a conventional manner used in this field. For example, the conjugation may be performed through an amidation reaction or a thioamidation reaction, but is not limited thereto.
또한 상기 접합은 통상적인 링커를 매개로 수행될 수 있으며, 예를 들어 상기 표지자에 링커를 미리 부착한 뒤, 상기 N-아실히드라존 유도체의 말단 아민기를 상기 표지자에 부착된 링커와 반응시킬 수 있다. 구체적으로, 상기 표지자로서 디페록사민(DFO)을 사용할 경우, 링커로서 p-NCS-벤젠이 부착된 화합물(p-NCS-Bz-DFO)을 반응에 투입할 수 있다.In addition, the conjugation can be carried out through a conventional linker, for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label. . Specifically, when deferoxamine (DFO) is used as the marker, a compound having p-NCS-benzene attached as a linker (p-NCS-Bz-DFO) may be added to the reaction.
또한 상기 접합은 통상적인 링커를 매개로 수행될 수 있으며, 예를 들어 상기 표지자에 링커를 미리 부착한 뒤, 상기 N-아실히드라존 유도체의 말단 아민기를 상기 표지자에 부착된 링커와 반응시킬 수 있다. 구체적으로, 표지자로서 디페록사민(DFO)을 사용할 경우, 링커로서 p-NCS-벤젠이 부착된 화합물(p-NCS-Bz-DFO)을 반응에 투입할 수 있다.In addition, the conjugation can be carried out through a conventional linker, for example, after the linker is attached to the label in advance, the terminal amine group of the N-acylhydrazone derivative can be reacted with the linker attached to the label. . Specifically, when deferoxamine (DFO) is used as a marker, a compound (p-NCS-Bz-DFO) to which p-NCS-benzene is attached as a linker may be added to the reaction.
또한 상기 표지자는 반응의 효율을 높이기 위해 말단이 개질된 것을 사용할 수 있는데, 예를 들어 표지자로서 시아닌 5(Cy5)를 사용할 경우 이를 N-히드록시석신이미드(NHS)와 에스터 결합하여 말단 개질한 뒤 반응에 투입할 수 있다.In addition, the marker may use a modified terminus in order to increase the efficiency of the reaction. For example, when cyanine 5 (Cy5) is used as a marker, it is terminally modified by ester bonding with N-hydroxysuccinimide (NHS). It can then be used in the reaction.
염 및 이성질체salts and isomers
본 발명은 상기 화학식 1의 N-아실히드라존 유도체 또는 상기 접합체의 약학적으로 허용가능한 염을 포함한다.The present invention includes an N-acylhydrazone derivative of Formula 1 or a pharmaceutically acceptable salt of the conjugate.
상기 약학적으로 허용가능한 염은 인간에 대한 독성이 낮아야 하며, 모 화합물의 생물학적 활성 및 물리화학적 특성에 임의의 부정적인 영향을 주지 않아야 한다. The pharmaceutically acceptable salt should have low toxicity to humans and should not adversely affect the biological activity and physicochemical properties of the parent compound.
예를 들어, 상기 약학적으로 허용가능한 염은 약학적으로 허용가능한 유리 산(free acid)에 의해 형성된 산부가염일 수 있다.For example, the pharmaceutically acceptable salt may be an acid addition salt formed with a pharmaceutically acceptable free acid.
상기 유리 산으로는 무기 산 또는 유기 산을 사용할 수 있으며, 이때 무기 산은 염산, 황산, 질산, 인산, 과염소산, 브롬산 등일 수 있고, 유기 산은 아세트산, 메탄설폰산, 에탄설폰산, p-톨루엔설폰산, 푸마르산, 말레산, 말론산, 프탈산, 석신산, 락트산, 시트르산, 글루콘산, 타르타르산, 살리실산, 말산, 옥살산, 벤조산, 엠본산, 아스파트산, 글루탐산 등일 수 있다. The free acid may be an inorganic acid or an organic acid, wherein the inorganic acid may be hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, hydrobromic acid, etc., and the organic acid is acetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid phonic acid, fumaric acid, maleic acid, malonic acid, phthalic acid, succinic acid, lactic acid, citric acid, gluconic acid, tartaric acid, salicylic acid, malic acid, oxalic acid, benzoic acid, embonic acid, aspartic acid, glutamic acid, and the like.
상기 산부가염은 통상의 방법, 예를 들어 모 화합물을 과량의 산 수용액에 용해시키고, 이 염을 수혼화성 유기 용매, 예를 들어 메탄올, 에탄올, 아세톤 또는 아세토니트릴을 사용하여 침전시켜서 제조될 수 있다. The acid addition salt can be prepared by a conventional method, for example, by dissolving the parent compound in an excess aqueous acid solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. .
또한, 상기 약학적으로 허용가능한 염은 알칼리금속염(나트륨염 등) 또는 알칼리토금속염(칼륨염 등)일 수 있다.In addition, the pharmaceutically acceptable salt may be an alkali metal salt (sodium salt, etc.) or alkaline earth metal salt (potassium salt, etc.).
상기 알칼리금속염 또는 알칼리토금속염은, 예를 들어 모 화합물을 과량의 알칼리금속 수산화물 또는 알칼리토금속 수산화물 용액 중에 용해시키고, 미용해된 화합물 염을 여과한 후 여액을 증발 및 건조시켜 얻을 수 있다. The alkali metal salt or alkaline earth metal salt can be obtained, for example, by dissolving the parent compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate.
또한, 상기 화학식 1의 N-아실히드라존 유도체 및 상기 접합체는 키랄 탄소 중심을 가질 수 있으며, 이에 따라 R 또는 S 이성질체, 라세미 화합물, 개개의 거울상 이성질체 또는 혼합물, 개개의 부분입체 이성질체 또는 혼합물 형태로 존재할 수 있으며, 이러한 모든 입체 이성질체 및 이들의 혼합물이 본 발명의 범주에 속할 수 있다.In addition, the N-acylhydrazone derivatives of formula (1) and the conjugates may have a chiral carbon center, and thus, in the form of R or S isomers, racemic compounds, individual enantiomers or mixtures, individual diastereomers or mixtures. may exist, and all such stereoisomers and mixtures thereof may fall within the scope of the present invention.
또한, 본 발명은 상기 화학식 1의 N-아실히드라존 유도체 또는 이와 표지자의 접합체의 수화물 및 용매화물을 포함할 수 있다. 상기 수화물 및 용매화물은 공지된 방법을 사용하여 제조될 수 있으며, 무독성 및 수용성인 것이 바람직하다. 특히, 바람직하게는 상기 수화물 및 용매화물은 각각 물 및 알코올성 용매(특히, 에탄올 등)의 1 내지 5개의 분자가 결합된 것일 수 있다.In addition, the present invention may include hydrates and solvates of the N-acylhydrazone derivative of Formula 1 or a conjugate thereof with a marker. The hydrates and solvates can be prepared using a known method, and preferably nontoxic and water-soluble. In particular, preferably, the hydrate and the solvate may be one in which 1 to 5 molecules of water and an alcoholic solvent (especially, ethanol, etc.) are bound, respectively.
따라서 이하에서 "본 발명에 따른 화합물"이라 하면, 상기 화학식 1의 N-아실히드라존 유도체 및 상기 접합체와 함께, 이들의 약학적으로 허용가능한 염, 입체이성질체, 수화물 및 용매화물을 모두 포함하는 범위로 해석될 수 있다.Therefore, in the following, when "a compound according to the present invention" is referred to as a "compound according to the present invention", the N-acylhydrazone derivative of Formula 1 and the conjugate, together with pharmaceutically acceptable salts, stereoisomers, hydrates and solvates thereof are included. can be interpreted as
본 발명에 따른 화합물의 용도Use of the compounds according to the invention
본 발명에 따른 상기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염은 암세포에 특이적으로 작용하여 축적되며, 항암 효능을 갖는 화합물을 기반으로 제조되어 치료와 진단이 모두 가능하고 안정성과 용해도가 우수하다.According to the present invention, the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer or a pharmaceutically acceptable salt thereof, acts specifically on cancer cells and accumulates, and is prepared based on a compound having anticancer efficacy Therefore, both treatment and diagnosis are possible, and it has excellent stability and solubility.
또한 상기 접합체에 적용되는 상기 화학식 1의 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염, 및 이를 포함하는 약학적 조성물은 미세소관의 튜불린 중합을 저해하여 세포자살을 유발하므로 암을 비롯한 세포증식성 질환의 예방 또는 치료에 유용하다.In addition, the N-acylhydrazone derivative of Formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same applied to the conjugate inhibit microtubule polymerization of tubulin to induce apoptosis. Therefore, it is useful for the prevention or treatment of cell proliferative diseases, including cancer.
구체적으로 상기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염은 미세소관의 튜불린 중합을 저해하여 세포자살을 유발하고 다중약물 내성을 보이는 암세포에도 효과적으로 작용하며, 암 전이 억제 활성도 우수하다. 또한 본 발명에 따른 화합물은 높은 체내 안정성 및 용해도를 나타내므로 생체이용률이 우수하다.Specifically, the conjugate of the N-acylhydrazone derivative of Formula 1 and the marker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof inhibits tubulin polymerization of microtubules to induce apoptosis and is also used in cancer cells exhibiting multidrug resistance. It works effectively and has excellent cancer metastasis inhibitory activity. In addition, since the compound according to the present invention exhibits high stability and solubility in the body, it has excellent bioavailability.
따라서 본 발명에 따른 화합물 또는 이를 포함하는 약학적 조성물은 세포증식성 질환의 진단과 함께 예방 및 치료에 사용될 수 있다. 또한, 본 발명에 따른 화합물은 미세소관을 탈중합시키는데 사용될 수 있다. 구체적으로, 본 발명에 따른 화합물은 튜불린을 저해, 보다 구체적으로 튜불린의 중합을 저해하는데 사용될 수 있다. 또한 본 발명에 따른 화합물은 튜불린의 콜히친 결합부위에 작용될 수 있고, 세포주기를 G2기 또는 M기에 정지시켜 세포자살을 유발할 수 있다. 또한 본 발명에 따른 화합물은 다중약물 내성을 보이는 암세포에 작용할 수 있다.Therefore, the compound according to the present invention or a pharmaceutical composition comprising the same can be used for diagnosis and prevention and treatment of cell proliferative diseases. In addition, the compounds according to the invention can be used to depolymerize microtubules. Specifically, the compound according to the present invention can be used to inhibit tubulin, more specifically, to inhibit polymerization of tubulin. In addition, the compound according to the present invention may act on the colchicine binding site of tubulin, and may induce apoptosis by stopping the cell cycle in the G2 or M phase. In addition, the compound according to the present invention can act on cancer cells exhibiting multidrug resistance.
예를 들어, 본 발명에 따른 화합물 또는 이를 포함하는 약학적 조성물은 세포증식성 질환의 발생, 진행 또는 전이의 진단제로 사용될 수 있다. 구체적으로, 본 발명에 따른 화합물 또는 이를 포함하는 약학적 조성물은 세포증식성 질환의 진단을 위한 조영제로 사용될 수 있다. 또한, 본 발명에 따른 화합물 또는 이를 포함하는 약학적 조성물은 튜불린 저해제 또는 항암제로 사용될 수 있다. 이에 따라 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 발생, 진행 또는 전이를 진단하기 위한 용도를 제공한다.For example, the compound according to the present invention or a pharmaceutical composition comprising the same can be used as a diagnostic agent for the occurrence, progression or metastasis of a cell proliferative disease. Specifically, the compound according to the present invention or a pharmaceutical composition comprising the same may be used as a contrast agent for the diagnosis of cell proliferative diseases. In addition, the compound according to the present invention or a pharmaceutical composition comprising the same may be used as a tubulin inhibitor or an anticancer agent. Accordingly, the present invention provides the use of a compound according to the present invention for diagnosing the occurrence, progression or metastasis of a cell proliferative disease.
또한 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 예방 또는 치료를 위한 용도를 제공한다. 또한 본 발명은, 본 발명에 따른 화합물의 튜불린의 중합을 저해하기 위한 용도를 제공한다. 또한 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 발생, 진행 또는 전이의 진단용 약제의 제조를 위한 용도를 제공한다. 또한 본 발명은, 세포증식성 질환의 발생, 진행 또는 전이의 진단에 사용하기 위한 본 발명에 따른 화합물을 포함하는 조성물을 제공한다. The present invention also provides the use of the compound according to the present invention for the prevention or treatment of a cell proliferative disease. The invention also provides the use of a compound according to the invention for inhibiting polymerization of tubulin. The present invention also provides the use of a compound according to the present invention for the manufacture of a medicament for diagnosis of the occurrence, progression or metastasis of a cell proliferative disease. The present invention also provides a composition comprising a compound according to the present invention for use in the diagnosis of the development, progression or metastasis of a cell proliferative disease.
또한 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 진단용 조영제의 제조를 위한 용도를 제공한다. 또한 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 발생, 진행 또는 전이의 진단용 약제의 제조를 위한 용도를 제공한다. 또한 본 발명은, 본 발명에 따른 화합물의 세포증식성 질환의 예방 또는 치료용 약제의 제조를 위한 용도를 제공한다. 또한 본 발명은, 본 발명에 따른 화합물의 튜불린 중합 저해용 약제의 제조를 위한 용도를 제공한다.The present invention also provides the use of the compound according to the present invention for the manufacture of a contrast agent for diagnosis of a cell proliferative disease. The present invention also provides the use of a compound according to the present invention for the manufacture of a medicament for diagnosis of the occurrence, progression or metastasis of a cell proliferative disease. The present invention also provides the use of the compound according to the present invention for the preparation of a medicament for preventing or treating a cell proliferative disease. The present invention also provides the use of the compound according to the present invention for the preparation of a medicament for inhibiting tubulin polymerization.
또한 본 발명은, 본 발명에 따른 화합물을 필요로 하는 대상에 투여하는 것을 포함하는, 세포증식성 질환의 발생, 진행 또는 전이를 진단하는 방법을 제공한다. 또한 본 발명은, 본 발명에 따른 화합물을 필요로 하는 대상에 투여하는 것을 포함하는, 세포증식성 질환을 갖는 부위를 이미지화하는 방법을 제공한다. 또한 본 발명은, 본 발명에 따른 화합물을 필요로 하는 대상에 투여하는 것을 포함하는, 튜불린의 중합을 저해하는 방법을 제공한다. 또한 본 발명은, 본 발명에 따른 화합물을 필요로 하는 대상에 투여하는 것을 포함하는, 세포증식성 질환을 예방 또는 치료하는 방법을 제공한다. The present invention also provides a method for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof. The present invention also provides a method of imaging a site having a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof. The present invention also provides a method for inhibiting polymerization of tubulin comprising administering to a subject in need thereof a compound according to the present invention. The present invention also provides a method for preventing or treating a cell proliferative disease, comprising administering a compound according to the present invention to a subject in need thereof.
여기서 "진단"이란 상기 화합물의 투여로 상기 질환의 발견 및 검출하거나 영상 장비를 이용하여 이미지화하는 모든 행위를 의미하고, "예방"이란 상기 화합물의 투여로 상기 질환의 발생, 확산 및 재발을 저해시키거나 지연시키는 모든 행위를 의미하고, "치료"는 상기 화합물의 투여로 상기 질환의 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.Here, "diagnosis" refers to any act of discovering and detecting the disease by administration of the compound or imaging using imaging equipment, and "prevention" refers to any activity that inhibits the occurrence, spread and recurrence of the disease by administration of the compound. It means any action that delays or improves, and "treatment" means any action in which the symptoms of the disease are ameliorated or beneficially changed by administration of the compound.
여기서 "필요로 하는 대상"이란, 상기 세포증식성 질환이 발병하였거나 발병할 수 있는 인간(환자)을 포함한 원숭이, 소, 말, 양, 돼지, 닭, 칠면조, 메추라기, 고양이, 개, 마우스, 쥐, 토끼, 기니아 피그 등의 모든 동물을 의미하며, 구체적으로 포유류를 의미할 수 있다. 또한, 상기 필요로 하는 대상은 생체 시료(biological sample)를 의미할 수도 있다.Herein, the term "subject in need" refers to monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, including humans (patients) who have or may develop the cell proliferative disease. It refers to all animals such as rabbits and guinea pigs, and may specifically refer to mammals. In addition, the required object may refer to a biological sample.
또한 "투여"란, 임의의 적절한 방법으로 이를 필요로 하는 대상에게 소정의 물질을 제공하는 것을 의미하며, 본 발명에 따른 화합물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. Also, "administration" means providing a predetermined substance to a subject in need thereof by any suitable method, and the administration route of the compound according to the present invention is administered through any general route as long as it can reach the target tissue. can be
약학적 조성물pharmaceutical composition
또한 본 발명은, 본 발명에 따른 화합물을 유효성분으로 포함하는 약학적 조성물을 제공한다. 또한 본 발명은, 본 발명에 따른 화합물을 유효성분으로 포함하는 튜불린 중합 저해를 위한 약학적 조성물을 제공한다. 또한 본 발명은, 본 발명에 따른 화합물을 유효성분으로 포함하는 세포증식성 질환의 발생, 진행 또는 전이를 진단하기 위한 약학적 조성물, 구체적으로 조영제를 제공한다. 상기 약학적 조성물은 세포증식성 질환의 치료 효과를 또한 나타낼 수 있다. 또한 본 발명은, 본 발명에 따른 화합물을 유효성분으로 포함하는 세포증식성 질환의 예방 또는 치료를 위한 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition comprising the compound according to the present invention as an active ingredient. The present invention also provides a pharmaceutical composition for inhibiting polymerization of tubulin comprising the compound according to the present invention as an active ingredient. The present invention also provides a pharmaceutical composition, specifically, a contrast agent for diagnosing the occurrence, progression or metastasis of a cell proliferative disease comprising the compound according to the present invention as an active ingredient. The pharmaceutical composition may also exhibit a therapeutic effect on cell proliferative diseases. The present invention also provides a pharmaceutical composition for the prevention or treatment of a cell proliferative disease comprising the compound according to the present invention as an active ingredient.
상기 세포증식성 질환은 암일 수 있다. 본 명세서에서 "암"은, 비제어된 방식으로 증식되거나 전이되는 경향이 있는 세포의 비정상적 성장을 지칭한다. 구체적으로 상기 암은 고형암, 혈액암, 또는 전이성암일 수 있다. 보다 구체적으로, 상기 암은 직장암, 유방암, 폐암, 위암, 간암, 백혈병, 신경교종, 피부암, 자궁경부암, 또는 이로부터 유래된 전이물일 수 있다. 따라서 본 발명에 따른 화합물을 포함하는 약학적 조성물은 상기 예시한 다양한 암의 진단제, 조영제, 치료제 또는 암 전이 억제제로 사용될 수 있다.The cell proliferative disease may be cancer. As used herein, “cancer” refers to the abnormal growth of cells that tend to proliferate or metastasize in an uncontrolled manner. Specifically, the cancer may be a solid cancer, a blood cancer, or a metastatic cancer. More specifically, the cancer may be rectal cancer, breast cancer, lung cancer, stomach cancer, liver cancer, leukemia, glioma, skin cancer, cervical cancer, or a metastasis derived therefrom. Accordingly, the pharmaceutical composition comprising the compound according to the present invention can be used as a diagnostic agent, contrast agent, therapeutic agent, or cancer metastasis inhibitor for various cancers exemplified above.
또한 본 발명은, 본 발명에 따른 화합물 및 약학적으로 허용가능한 첨가제를 포함하는 약학적 조성물을 제공한다. 본 발명의 약학적 조성물은 유효성분으로서 본 발명에 따른 화합물을, 약학적 조성물의 총 중량을 기준으로 0.1 중량% 내지 90 중량%, 구체적으로 0.1 중량% 내지 75 중량%, 보다 구체적으로 1 중량% 내지 50 중량%로 함유할 수 있다.The present invention also provides a pharmaceutical composition comprising the compound according to the present invention and a pharmaceutically acceptable additive. The pharmaceutical composition of the present invention contains the compound according to the present invention as an active ingredient, 0.1 wt% to 90 wt%, specifically 0.1 wt% to 75 wt%, more specifically 1 wt%, based on the total weight of the pharmaceutical composition to 50% by weight.
본 발명의 약학적 조성물은, 통상적인 방법에 따라 통상적이고 무독성인 약학적으로 허용가능한 첨가제를 포함할 수 있다. 예를 들어, 상기 약학적 조성물은 약학적으로 허용되는 담체, 희석제 또는 부형제를 추가로 포함할 수 있다.The pharmaceutical composition of the present invention may include conventional and non-toxic pharmaceutically acceptable excipients according to conventional methods. For example, the pharmaceutical composition may further include a pharmaceutically acceptable carrier, diluent or excipient.
본 발명의 약학적 조성물에 사용되는 첨가제의 예는 감미제, 결합제, 용매, 용해 보조제, 습윤제, 유화제, 등장화제, 흡수제, 붕해제, 산화방지제, 보존제, 윤활제, 충전제, 향미제 등을 포함할 수 있다. 예를 들어, 상기 첨가제는 락토오스, 덱스트로스, 수크로스, 만니톨, 소르비톨, 셀룰로스, 글리신, 실리카, 활석, 스테아르산, 스테아린, 마그네슘 스테아레이트, 마그네슘 알루미노실리케이트, 전분, 젤라틴, 트라가칸트 검, 알긴산, 나트륨 알기네이트, 메틸셀룰로스, 나트륨 카복시메틸셀룰로스, 한천, 물, 에탄올, 폴리에틸렌 글리콜, 폴리비닐피롤리돈, 염화나트륨, 염화칼슘, 오렌지 에센스, 딸기 에센스, 바닐라 향 등을 포함할 수 있다.Examples of additives used in the pharmaceutical composition of the present invention may include sweeteners, binders, solvents, solubilizing agents, wetting agents, emulsifiers, isotonic agents, absorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, flavoring agents, and the like. have. For example, the additive may be lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, stearin, magnesium stearate, magnesium aluminosilicate, starch, gelatin, gum tragacanth, alginic acid, sodium alginate, methylcellulose, sodium carboxymethylcellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavor, and the like.
본 발명의 약학적 조성물은 경구 투여(예컨대, 정제, 환제, 산제, 캡슐제, 시럽 또는 에멀젼) 또는 비경구 투여(예컨대, 근육내, 정맥내 또는 피하 주사)를 위한 다양한 제제 형태로 배합될 수 있다. The pharmaceutical composition of the present invention may be formulated in various formulations for oral administration (eg, tablets, pills, powders, capsules, syrups or emulsions) or parenteral administration (eg, intramuscular, intravenous or subcutaneous injection). have.
바람직하게는 본 발명의 약학적 조성물은 경구 투여용 제제로 배합될 수 있으며, 이때 사용되는 첨가제로는 셀룰로스, 칼슘 실리케이트, 옥수수 전분, 락토오스, 수크로스, 덱스트로스, 칼슘 포스페이트, 스테아르산, 마그네슘 스테아레이트, 칼슘 스테아레이트, 젤라틴, 활석, 계면활성제, 현탁제, 유화제, 희석제 등이 포함될 수 있다.Preferably, the pharmaceutical composition of the present invention may be formulated as a formulation for oral administration, and the additives used in this case include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, and magnesium stearate. lactate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifying agents, diluents, and the like may be included.
구체적으로, 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 약학적 조성물에 적어도 하나 이상의 부형제, 예를 들면 전분, 탄산칼슘, 수크로스, 락토즈, 젤라틴 등을 혼합하여 제제화될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크와 같은 윤활제가 사용될 수 있다.Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the pharmaceutical composition, for example, starch, calcium carbonate, sucrose, It may be formulated by mixing lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may be used.
또한, 경구 투여를 위한 액상 제제로는 현탁제, 유제, 시럽제 등이 예시될 수 있으며, 흔히 사용되는 단순 희석제인 물, 액체 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.In addition, liquid formulations for oral administration may be exemplified by suspensions, emulsions, syrups, etc., and various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are commonly used simple diluents. may be included.
또한, 비경구 투여를 위한 제제에는 멸균된 수용액제, 비수성용제, 현탁제, 유제, 동결건조 제제 및 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로골, 트윈61. 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. 한편, 주사제에는 용해제, 등장화제, 현탁화제, 유화제, 안정화제, 방부제 등과 같은 종래의 첨가제가 포함될 수 있다.In addition, preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. The suppositories are Witepsol, Macrogol, and Twin61. Cacao butter, laurin fat, glycerogelatin, etc. may be used. On the other hand, the injection may contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, and the like.
또한, 상기 약학적 조성물이 세포증식성 질환의 진단을 위한 용도, 예를 들어 형광 조영제로 사용될 경우, 본 발명에 따른 화합물을 주사용 증류수, 약리 식염수, 링거 용액 등과 같이 용매 중에 현탁되거나 용해하여 포함할 수 있다. 필요한 경우, 담체, 부형제 등과 같은 제약상 허용가능한 첨가제가 첨가될 수 있다. 이러한 첨가제는 약리학적으로 허용가능한 전해질, 완충액, 세정제 및 삼투압 조절용 물질 및 안정성과 용해도를 개선시키는 물질(예를 들어, 시클로덱스트린, 리포솜 등)을 포함한다. 또한 관련 분야에서 일반적으로 사용되는 다양한 첨가제가 사용될 수 있다. In addition, when the pharmaceutical composition is used for the diagnosis of cell proliferative diseases, for example, as a fluorescent contrast agent, the compound according to the present invention may be suspended or dissolved in a solvent such as distilled water for injection, pharmacological saline, Ringer's solution, etc. can If necessary, pharmaceutically acceptable additives such as carriers, excipients and the like may be added. Such additives include pharmacologically acceptable electrolytes, buffers, detergents and substances for regulating osmotic pressure and substances for improving stability and solubility (eg, cyclodextrins, liposomes, etc.). In addition, various additives commonly used in the related field may be used.
또한 상기 약학적 조성물이 방사성 진단을 위한 용도, 예를 들어 양성자방출 단층촬영(PET) 등을 위한 방사성 조영제로 사용될 경우, 주사제로서 제제화될 수 있으며, 보다 구체적으로 정맥주사제로 제제화될 수 있다. 주사제로 제제화될 경우 상기 약학적 조성물은 혈액과 등장인 무독성 완충용액을 희석제로서 포함할 수 있으며, 예를 들어 pH 7.4의 인산완충용액 등을 포함할 수 있고, 완충용액 이외에 기타 다른 희석제 또는 첨가제도 더 포함할 수 있다. 이러한 주사제에 부가될 수 있는 부형제 및 첨가제는 당해 기술분야에서 통상의 지식을 가진 자에게 널리 공지되어 있다.In addition, when the pharmaceutical composition is used as a radioactive contrast agent for radiodiagnosis, for example, proton emission tomography (PET), it may be formulated as an injection, and more specifically, it may be formulated as an intravenous injection. When formulated as an injection, the pharmaceutical composition may include a non-toxic buffer solution isotonic with blood as a diluent, and may include, for example, a phosphate buffer solution of pH 7.4, and other diluents or additives in addition to the buffer solution. may include more. Excipients and additives that can be added to such injections are well known to those of ordinary skill in the art.
본 발명에 따른 화합물 또는 약학적 조성물은 치료학적으로 유효한 양 또는 약학적으로 유효한 양으로 환자에 투여될 수 있다. The compound or pharmaceutical composition according to the present invention may be administered to a patient in a therapeutically effective amount or in a pharmaceutically effective amount.
여기서 "치료학적으로 유효한 양" 또는 "약학적으로 유효한 양"이란 대상 질환을 예방 또는 치료하는데 유효한 화합물 또는 약학적 조성물의 양으로서, 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분하며 부작용을 일으키지 않을 정도의 양을 의미한다. 상기 유효량의 수준은 환자의 건강상태, 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 방법, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 배합 또는 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 또한 상기 "약학적으로 유효한 양"은 대상 질환을 진단하는데 유효한 화합물 또는 약학적 조성물의 양을 의미할 수 있다.Herein, "therapeutically effective amount" or "pharmaceutically effective amount" refers to an amount of a compound or pharmaceutical composition effective to prevent or treat a target disease, and is used to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment. It is sufficient and refers to an amount sufficient to not cause side effects. The level of the effective amount may be determined by the patient's health condition, disease type, severity, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment period, factors including the combination or concurrently used drugs; It may be determined according to factors well known in the medical field. In addition, the "pharmaceutically effective amount" may refer to an amount of a compound or pharmaceutical composition effective for diagnosing a target disease.
본 발명에 따른 화합물 또는 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고, 종래의 치료제와 순차적으로 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The compound or pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.
구체적으로, 본 발명의 약학적 조성물에서 화합물의 유효량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으며, 일반적으로는 체중 1 kg 당 0.1 mg 내지 100 mg, 또는 0.5 mg 내지 10 mg을 매일 또는 격일 투여하거나 1일 2회 또는 3회로 나누어 투여할 수 있다. 그러나, 투여 경로, 질병의 중증도, 성별, 체중, 연령 등에 따라서 증감될 수 있으므로, 본 발명의 범위는 이에 한정되지 않는다.Specifically, the effective amount of the compound in the pharmaceutical composition of the present invention may vary depending on the age, sex, and weight of the patient, and in general, 0.1 mg to 100 mg, or 0.5 mg to 10 mg per kg of body weight, daily or every other day It can be administered or divided into 2 or 3 times a day. However, the scope of the present invention is not limited thereto since it may increase or decrease depending on the route of administration, disease severity, sex, weight, age, etc.
또한 본 발명의 약학적 조성물을 형광 조영제로서 사용할 경우, 주사, 분무 또는 도포, 정맥내(정맥, 동맥), 경구, 복강내, 경피, 피하내, 낭내 또는 기관지내로 생체에 투여될 수 있다. 상기 약학적 조성물의 투여량은 본 발명에 따른 화합물의 양으로 체중 1 kg 당 0.1 mg 내지 100 mg, 바람직하게는 0.5 mg 내지 20 mg이며, 사용되는 형광을 방출하는 화합물의 종류, 피검자의 나이, 체중 및 표적 기관 등에 따라 적절하게 조절될 수 있다. 또한 본 발명의 약학적 조성물을 방사성 진단을 위한 용도, 예를 들어 PET 조영제로 사용할 경우, 유효성분인 본 발명에 따른 화합물을 기준으로 인간 남성 성인을 기준으로 약 1 mCi 내지 3 mCi의 양을 투여할 수 있으며, 인종, 성별, 나이, 체중, 염증의 종류 등에 따라 전문 의사가 적절히 증감할 수 있다.In addition, when the pharmaceutical composition of the present invention is used as a fluorescent contrast agent, it can be administered to a living body by injection, spraying or application, intravenous (venous, arterial), oral, intraperitoneal, transdermal, subcutaneous, intracystic or intrabronchial. The dosage of the pharmaceutical composition is 0.1 mg to 100 mg, preferably 0.5 mg to 20 mg per 1 kg of body weight, in the amount of the compound according to the present invention, the type of compound emitting fluorescence used, the age of the subject, It can be appropriately adjusted according to body weight and target organs. In addition, when the pharmaceutical composition of the present invention is used for radiodiagnosis, for example, as a PET contrast agent, an amount of about 1 mCi to 3 mCi is administered based on a human male adult based on the compound according to the present invention as an active ingredient. Depending on race, gender, age, weight, type of inflammation, etc., a professional doctor can appropriately increase or decrease it.
본 발명에 따른 화합물 또는 약학적 조성물은 화학요법, 방사선 요법, 면역요법, 호르몬 치료, 골수 이식, 줄기세포 대체치료, 다른 생물학적 치료, 수술적 개입 또는 이들의 조합과 병용하여 종양 요법을 위해 투여될 수 있다. 예컨대, 본 발명에 따른 화합물 또는 약학적 조성물은 장기적으로 진행되는 다른 치료 전략과 함께 보조 요법으로 사용되거나, 중증 환자에서 종양 퇴행 또는 화학 예방 요법 후 환자의 상태를 유지하기 위해 사용될 수 있다.The compound or pharmaceutical composition according to the present invention may be administered for tumor therapy in combination with chemotherapy, radiation therapy, immunotherapy, hormone therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapy, surgical intervention or a combination thereof. can For example, the compound or pharmaceutical composition according to the present invention can be used as an adjuvant therapy in combination with other long-term treatment strategies, or to maintain the patient's condition after tumor regression or chemopreventive therapy in seriously ill patients.
바람직하게는, 본 발명의 약학적 조성물은 1종 이상의 유효성분을 추가로 포함할 수 있으며, 상기 추가의 유효성분은 항-증식 화합물, 예컨대, 아로마타제 저해제, 항-에스트로겐, 토포이소머라제 I 저해제, 토포이소머라제 II 저해제, 미세소관 활성 화합물, 알킬화 화합물, 히스톤 데아세틸라제 저해제, 세포 분화 과정을 유도하는 화합물, 시클로옥시게나제 저해제, MMP 저해제, mTOR 저해제, 항-신생물 항-대사물질, 플라틴 화합물, 단백질 또는 지질 키나제 활성을 표적화/감소시키는 화합물, 항-혈관신생 화합물, 단백질 또는 지질 포스파타제의 활성을 표적화하거나 감소시키거나 저해하는 화합물, 고나도렐린 효능제, 항-안드로겐, 메티오닌 아미노펩티다제 저해제, 비스포스포네이트, 생물학적 반응 개질제, 항-증식성 항체, 헤파라나제 저해제, Ras 종양원성 이소형의 저해제, 텔로머라제 저해제, 프로테아솜 저해제, 혈액계 악성종양의 치료에 사용되는 화합물, Flt-3의 활성을 표적화하거나 감소시키거나 저해하는 화합물, Hsp90 저해제, 키네신 스핀들 단백질 저해제, MEK 저해제, 류코보린, EDG 결합제, 항-백혈병 화합물, 리보뉴클레오티드 리덕타제 저해제, S-아데노실메티오닌 데카르복실라제 저해제, 지혈성 스테로이드, 코르티코스테로이드, 다른 화학요법 화합물, 광감작 화합물일 수 있으나, 이에 제한되지 않는다.Preferably, the pharmaceutical composition of the present invention may further include one or more active ingredients, wherein the additional active ingredient is an anti-proliferative compound, such as an aromatase inhibitor, anti-estrogens, topoisomerase I Inhibitors, topoisomerase II inhibitors, microtubule active compounds, alkylation compounds, histone deacetylase inhibitors, compounds inducing cellular differentiation processes, cyclooxygenase inhibitors, MMP inhibitors, mTOR inhibitors, anti-neoplastic anti-metabolism Substances, platin compounds, compounds targeting/reducing protein or lipid kinase activity, anti-angiogenic compounds, compounds targeting, reducing or inhibiting the activity of protein or lipid phosphatase, gonadorelin agonists, anti-androgens, Methionine aminopeptidase inhibitors, bisphosphonates, biological response modifiers, anti-proliferative antibodies, heparanase inhibitors, inhibitors of Ras tumorigenic isoforms, telomerase inhibitors, proteasome inhibitors, used in the treatment of hematologic malignancies compounds targeting, reducing or inhibiting the activity of Flt-3, Hsp90 inhibitors, kinesin spindle protein inhibitors, MEK inhibitors, leucovorin, EDG binding agents, anti-leukemia compounds, ribonucleotide reductase inhibitors, S-adenosyl It may be, but is not limited to, a methionine decarboxylase inhibitor, a hemostatic steroid, a corticosteroid, another chemotherapeutic compound, or a photosensitizer compound.
이하, 본 발명을 하기 실시예에 의하여 더욱 구체적으로 설명한다. 그러나 하기의 실시예는 본 발명을 더욱 쉽게 이해하기 위하여 제공되는 것일 뿐, 실시예에 의하여 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only provided for easier understanding of the present invention, and the content of the present invention is not limited by the examples.
하기 실시예에서 사용된 약어의 정의는 다음과 같다:Definitions of abbreviations used in the examples below are as follows:
EA: 에틸 아세테이트, EtOH: 에탄올, EA: ethyl acetate, EtOH: ethanol,
DMF: 디메틸폼아미드, n-Hex: n-헥산, DMF: dimethylformamide, n-Hex: n-hexane,
PrOH: 프로판올, DMSO: 디메틸설폭시드.PrOH: propanol, DMSO: dimethyl sulfoxide.
<방향족 치환된 벤조퓨란-2-카보히드라지드 유도체의 제조><Preparation of aromatic substituted benzofuran-2-carbohydrazide derivative>
Figure PCTKR2021007223-appb-img-000041
Figure PCTKR2021007223-appb-img-000041
제조예 1: 에틸 5-메틸벤조퓨란-2-카복실레이트Preparation Example 1: Ethyl 5-methylbenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000042
Figure PCTKR2021007223-appb-img-000042
5-메틸살리실알데히드(1.40 g, 10.3 mmol)를 DMF(30 mL)에 녹인 후, 여기에 Ce2SO4(10.07 g, 30.9 mmol)와 에틸 브로모아세테이트(1.38 mL, 12.4 mmol)를 넣고 70℃에서 15 시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=10:1)로 분리하여 표제 화합물 1.28 g(수율 61%, 무색 액체)을 얻었다. After 5-methylsalicylaldehyde (1.40 g, 10.3 mmol) was dissolved in DMF (30 mL), Ce 2 SO 4 (10.07 g, 30.9 mmol) and ethyl bromoacetate (1.38 mL, 12.4 mmol) were added thereto. The reaction was carried out at 70°C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=10:1) to obtain 1.28 g (yield 61%, colorless liquid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 7.49-7.46 (m, 3H), 7.26 (m, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.46 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 159.85, 154.37, 145.92, 133.51, 129.27, 127.22, 122.45, 113.74, 112.02, 61.61, 21.45, 14.51; mp 38-39℃ 1 H NMR (400 MHz, CDCl 3 ): δ 7.49-7.46 (m, 3H), 7.26 (m, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.46 (s, 3H), 1.44 (t) , J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 159.85, 154.37, 145.92, 133.51, 129.27, 127.22, 122.45, 113.74, 112.02, 61.61, 21.45, 14.51; mp 38-39℃
제조예 2: 5-메틸벤조퓨란-2-카보히드라지드Preparation Example 2: 5-methylbenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000043
Figure PCTKR2021007223-appb-img-000043
상기 제조예 1에서 얻은 에틸 5-메틸벤조퓨란-2-카복실레이트(2.04 g, 10.0 mmol)를 EtOH(30 mL)에 녹이고 여기에 히드라진 일수화물(1.50 g, 30.0 mmol)을 넣어 24시간 환류시켰다. 반응용액을 감압증류하고 얻어진 고체를 물로 세척한 다음, 건조하여 표제 화합물 1.72 g(수율 90%, 흰색 고체)을 얻었다. Ethyl 5-methylbenzofuran-2-carboxylate (2.04 g, 10.0 mmol) obtained in Preparation Example 1 was dissolved in EtOH (30 mL), hydrazine monohydrate (1.50 g, 30.0 mmol) was added thereto, and refluxed for 24 hours. . The reaction solution was distilled under reduced pressure, and the resulting solid was washed with water and dried to obtain 1.72 g (yield 90%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 9.97 (s, 1H), 7.52-7.49 (m, 2H), 7.43 (s, 1H), 7.25 (dd, J = 8.4, 1.6 Hz, 1H), 4.55 (br s, 2H), 2.40 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 157.92, 152.64, 148.48, 132.68, 127.84, 127.11, 122.04, 111.27, 108.47, 20.81; MS (MALDI-TOF): m/z 213 [M+Na]+; mp 159℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.97 (s, 1H), 7.52-7.49 (m, 2H), 7.43 (s, 1H), 7.25 (dd, J = 8.4, 1.6 Hz, 1H) , 4.55 (br s, 2H), 2.40 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 157.92, 152.64, 148.48, 132.68, 127.84, 127.11, 122.04, 111.27, 108.47, 20.81; MS (MALDI-TOF): m/z 213 [M+Na] + ; mp 159℃
제조예 3: 에틸 5-메톡시벤조퓨란-2-카복실레이트Preparation Example 3: Ethyl 5-methoxybenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000044
Figure PCTKR2021007223-appb-img-000044
2-히드록시-5-메톡시벤즈알데히드(1.25 mL, 10.0 mmol)를 DMF(30 mL)에 녹인 후, 여기에 K2CO3(6.91 g, 50.0 mmol)와 에틸 브로모아세테이트(1.33 mL, 12.0 mmol)를 넣고 70℃에서 15시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=3:1)로 분리하여 표제 화합물 1.1 g(수율 50%, 흰색 고체)을 얻었다. After dissolving 2-hydroxy-5-methoxybenzaldehyde (1.25 mL, 10.0 mmol) in DMF (30 mL), K 2 CO 3 (6.91 g, 50.0 mmol) and ethyl bromoacetate (1.33 mL, 12.0) mmol) and reacted at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=3:1) to obtain 1.1 g of the title compound (yield 50%, white solid).
1H NMR (400 MHz, CDCl3): δ 7.50-7.47 (m, 2H), 7.08-7.05 (m, 2H), 4.45 (q, 2H, J = 7.2 Hz), 3.86 (s, 3H), 1.44 (t, 3H, J = 7.2 Hz); mp 53℃ 1 H NMR (400 MHz, CDCl 3 ): δ 7.50-7.47 (m, 2H), 7.08-7.05 (m, 2H), 4.45 (q, 2H, J = 7.2 Hz), 3.86 (s, 3H), 1.44 (t, 3H, J = 7.2 Hz); mp 53℃
제조예 4: 5-메톡시벤조퓨란-2-카보히드라지드Preparation Example 4: 5-Methoxybenzofuran-2-Carbohydrazide
Figure PCTKR2021007223-appb-img-000045
Figure PCTKR2021007223-appb-img-000045
상기 제조예 3에서 얻은 에틸 5-메톡시벤조퓨란-2-카복실레이트(1.03 g, 4.68 mmol)를 EtOH(30 mL)에 녹이고 여기에 히드라진 일수화물(702.8 mg, 14.04 mmol)을 넣어 24시간 환류시켰다. 반응용액을 감압증류하고 얻어진 고체를 CH2Cl2로 세척한 다음, 건조하여 표제 화합물 918 mg(수율 95%, 흰색 고체)을 얻었다. Ethyl 5-methoxybenzofuran-2-carboxylate (1.03 g, 4.68 mmol) obtained in Preparation Example 3 was dissolved in EtOH (30 mL), hydrazine monohydrate (702.8 mg, 14.04 mmol) was added thereto, and refluxed for 24 hours. made it The reaction solution was distilled under reduced pressure, and the resulting solid was washed with CH 2 Cl 2 and dried to obtain 918 mg (yield 95%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 9.98 (s, 1H), 7.53 (d, 1H, J = 9.0 Hz), 7.44 (s, 1H), 7.25 (d, 1H, J = 2.7 Hz), 7.02 (dd, 1H, J = 9.0, 2.7 Hz), 4.55 (br s, 2H), 3.79 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 157.85, 155.97, 149.10, 149.03, 127.65, 115.80, 112.34, 108.34, 104.09, 55.58; MS (MALDI-TOF): m/z 229.0 [M+Na]+, 245 [M+K]+; mp 163-164℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.98 (s, 1H), 7.53 (d, 1H, J = 9.0 Hz), 7.44 (s, 1H), 7.25 (d, 1H, J = 2.7 Hz) ), 7.02 (dd, 1H, J = 9.0, 2.7 Hz), 4.55 (br s, 2H), 3.79 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 157.85, 155.97, 149.10, 149.03, 127.65, 115.80, 112.34, 108.34, 104.09, 55.58; MS (MALDI-TOF): m/z 229.0 [M+Na] + , 245 [M+K] + ; mp 163-164℃
제조예 5: 5-클로로벤조퓨란-2-카보히드라지드Preparation 5: 5-chlorobenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000046
Figure PCTKR2021007223-appb-img-000046
0℃에서 CH2Cl2(5 mL)에 옥살릭클로라이드(257.2 μL, 3.0 mmol)와 DMF(50 μL)를 넣고 5분 교반한 다음, 5-클로로벤조퓨란-2-카복실산(393.2 mg, 2.0 mmol)의 CH2Cl2/DMF(5 mL, 4:1)의 혼합용액을 넣고 실온에서 2시간 교반하였다. 반응용액을 감압 증류하고 다시 CH2Cl2(5 mL)를 넣은 다음, 0℃에서 히드라진 일수화물(120.1 mg, 2.4 mmol)의 CH2Cl2(5 mL) 용액과 N,N-다이아이소프로필에틸아민(DIEA, 1.74 mL, 10 mmol)를 넣고 실온에서 15시간 교반하였다. 반응용액을 물로 희석시킨 후, EA로 추출하였다. 유기층을 Na2SO4로 건조, 여과하고 감압 증류하였다. 잔류물을 CH2Cl2로 세척하고 건조하여 표제 화합물 295 mg(수율 70%, 연노란색 고체)을 얻었다. At 0 °C, CH 2 Cl 2 (5 mL) was added with oxalic chloride (257.2 μL, 3.0 mmol) and DMF (50 μL), stirred for 5 minutes, and then 5-chlorobenzofuran-2-carboxylic acid (393.2 mg, 2.0 mmol) of CH 2 Cl 2 /DMF (5 mL, 4:1) was added and stirred at room temperature for 2 hours. The reaction solution was distilled under reduced pressure, and CH 2 Cl 2 (5 mL) was added thereto, and then a CH 2 Cl 2 (5 mL) solution of hydrazine monohydrate (120.1 mg, 2.4 mmol) and N,N-diisopropyl at 0° C. Ethylamine (DIEA, 1.74 mL, 10 mmol) was added and stirred at room temperature for 15 hours. The reaction solution was diluted with water and then extracted with EA. The organic layer was dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was washed with CH 2 Cl 2 and dried to obtain 295 mg (yield 70%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 10.10 (s, 1H), 7.86 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 9.2 Hz, 1H), 7.48 (s, 1H), 7.46 (dd, J = 9.2, 2.4 Hz, 1H), 4.59 (br s, 2H); 13C NMR (100 MHz, DMSO-d6): δ 157.41, 152.66, 149.79, 128.65, 127.99, 126.56, 122.02, 113.44, 108.25; MS (MALDI-TOF): m/z 233 [M+Na]+; mp 174-175℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.10 (s, 1H), 7.86 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 9.2 Hz, 1H), 7.48 (s, 1H) ), 7.46 (dd, J = 9.2, 2.4 Hz, 1H), 4.59 (br s, 2H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 157.41, 152.66, 149.79, 128.65, 127.99, 126.56, 122.02, 113.44, 108.25; MS (MALDI-TOF): m/z 233 [M+Na] + ; mp 174-175℃
제조예 6: 에틸 4,7-디메틸벤조퓨란-2-카복실레이트Preparation 6: Ethyl 4,7-dimethylbenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000047
Figure PCTKR2021007223-appb-img-000047
3,6-디메틸살리실알데히드(450.5 mg, 3.0 mmol)를 DMF(30 mL)에 녹인 후, 여기에 Ce2SO4(2.93 g, 9.0 mmol)와 에틸 브로모아세테이트(399.2 μL, 3.60 mmol)를 넣고 70℃에서 15시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=20:1)로 분리하여 표제 화합물 360 mg(수율 55%, 무색 액체)을 얻었다. 3,6-dimethylsalicylaldehyde (450.5 mg, 3.0 mmol) was dissolved in DMF (30 mL), and then Ce 2 SO 4 (2.93 g, 9.0 mmol) and ethyl bromoacetate (399.2 μL, 3.60 mmol) was added and reacted at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=20:1) to obtain 360 mg (yield 55%, colorless liquid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 7.56 (s, 1H), 7.14 (d, J = 7.2 Hz, 1H), 6.99 (d, J = 7.2 Hz, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.55 (s, 3H), 2.52 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 159.91, 154.90, 145.08, 130.19, 128.30, 126.63, 123.90, 119.85, 113.01, 61.43, 18.29, 15.00, 14.46 1 H NMR (400 MHz, CDCl 3 ): δ 7.56 (s, 1H), 7.14 (d, J = 7.2 Hz, 1H), 6.99 (d, J = 7.2 Hz, 1H), 4.45 (q, J = 7.2) Hz, 2H), 2.55 (s, 3H), 2.52 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 159.91, 154.90, 145.08, 130.19, 128.30, 126.63, 123.90, 119.85, 113.01, 61.43, 18.29, 15.00, 14.46
제조예 7: 4,7-디메틸벤조퓨란-2-카보히드라지드Preparation Example 7: 4,7-dimethylbenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000048
Figure PCTKR2021007223-appb-img-000048
상기 제조예 6에서 얻은 에틸 4,7-디메틸벤조퓨란-2-카복실레이트(290.3 mg, 1.33 mmol)를 EtOH(20 mL)에 녹이고 여기에 히드라진 일수화물(199.7 mg, 3.99 mmol)을 넣어 24시간 환류시켰다. 반응용액을 감압증류하고 얻어진 고체를 물로 세척한 다음, 건조하여 표제 화합물 236 mg(수율 87%, 흰색 고체)을 얻었다. Ethyl 4,7-dimethylbenzofuran-2-carboxylate (290.3 mg, 1.33 mmol) obtained in Preparation Example 6 was dissolved in EtOH (20 mL), and hydrazine monohydrate (199.7 mg, 3.99 mmol) was added thereto for 24 hours. refluxed. The reaction solution was distilled under reduced pressure, and the obtained solid was washed with water and dried to obtain 236 mg (yield 87%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 9.97 (s, 1H), 7.55 (s, 1H), 7.13 (d, J = 7.4 Hz, 1H), 7.00 (d, J = 7.4 Hz, 1H), 4.59 (br s, 2H), 2.46 (s, 6H, CH3x2); 13C NMR (100 MHz, DMSO-d6): δ 158.05, 153.07, 147.65, 129.34, 127.07, 126.47, 123.64, 118.65, 107.94, 17.86, 14.38; MS (MALDI-TOF): m/z 227 [M+Na]+; mp 187℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.97 (s, 1H), 7.55 (s, 1H), 7.13 (d, J = 7.4 Hz, 1H), 7.00 (d, J = 7.4 Hz, 1H) ), 4.59 (br s, 2H), 2.46 (s, 6H, CH 3 x2); 13 C NMR (100 MHz, DMSO-d 6 ): δ 158.05, 153.07, 147.65, 129.34, 127.07, 126.47, 123.64, 118.65, 107.94, 17.86, 14.38; MS (MALDI-TOF): m/z 227 [M+Na] + ; mp 187℃
제조예 8: 에틸 4,6-디메톡시벤조퓨란-2-카복실레이트Preparation 8: Ethyl 4,6-dimethoxybenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000049
Figure PCTKR2021007223-appb-img-000049
4,6-디메톡시살리실알데히드(499.2 mg, 2.74 mmol)를 DMF(10 mL)에 녹인 후, 여기에 K2CO3(1.89 g, 13.7 mmol)와 에틸 브로모아세테이트(364.8 μL, 3.29 mmol)를 넣고 70℃에서 15시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=20:1)로 분리하여 표제 화합물 460 mg(수율 67%, 흰색 고체)을 얻었다. 4,6-dimethoxysalicylaldehyde (499.2 mg, 2.74 mmol) was dissolved in DMF (10 mL), and then K 2 CO 3 (1.89 g, 13.7 mmol) and ethyl bromoacetate (364.8 μL, 3.29 mmol) were dissolved therein. ) and reacted at 70 °C for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=20:1) to obtain 460 mg (yield 67%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 7.54 (m, 1H), 6.67 (m, 1H), 6.35 (d, J = 1.9 Hz, 1H), 4.41 (q, J = 7.2 Hz, 2H), 3.90 (s, 3H), 3.857 (s, 3H), 1.40 (t, J = 7.2 Hz, 3H); (MALDI-TOF): m/z 251 [M+H]+; mp 98℃ 1 H NMR (400 MHz, CDCl 3 ): δ 7.54 (m, 1H), 6.67 (m, 1H), 6.35 (d, J = 1.9 Hz, 1H), 4.41 (q, J = 7.2 Hz, 2H), 3.90 (s, 3H), 3.857 (s, 3H), 1.40 (t, J = 7.2 Hz, 3H); (MALDI-TOF): m/z 251 [M+H] + ; mp 98℃
제조예 9: 4,6-디메톡시벤조퓨란-2-카보히드라지드Preparation Example 9: 4,6-dimethoxybenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000050
Figure PCTKR2021007223-appb-img-000050
상기 제조예 8에서 얻은 에틸 4,6-디메톡시벤조퓨란-2-카복실레이트(450.5 mg, 1.80 mmol)를 EtOH(20 mL)에 녹이고 여기에 히드라진 일수화물(450.5 mg, 9.0 mmol)을 넣어 6시간 환류시켰다. 반응용액을 차가운 물에 넣고 생성된 고체를 여과한 다음, 에틸 에테르로 세척하고 건조하여 표제 화합물 360 mg(수율 85%, 흰색 고체)을 얻었다. Ethyl 4,6-dimethoxybenzofuran-2-carboxylate (450.5 mg, 1.80 mmol) obtained in Preparation Example 8 was dissolved in EtOH (20 mL), and hydrazine monohydrate (450.5 mg, 9.0 mmol) was added thereto. time was refluxed. The reaction solution was poured into cold water, the resulting solid was filtered, washed with ethyl ether, and dried to obtain 360 mg of the title compound (yield 85%, white solid).
1H NMR (400 MHz, DMSO-d6): δ 9.78 (s, 1H), 7.40 (d, J = 0.8 Hz, 1H), 6.79 (m, 1H), 6.45 (d, J = 2.0 Hz, 1H), 4.49 (br s, 2H), 3.88 (s, 3H), 3.82 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 160.64, 158.10, 156.16, 153.94, 146.15, 110.83, 105.97, 94.98, 88.36, 55.79, 55.71; MS (MALDI-TOF): m/z 259 [M+Na]+; mp 194℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.78 (s, 1H), 7.40 (d, J = 0.8 Hz, 1H), 6.79 (m, 1H), 6.45 (d, J = 2.0 Hz, 1H) ), 4.49 (br s, 2H), 3.88 (s, 3H), 3.82 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 160.64, 158.10, 156.16, 153.94, 146.15, 110.83, 105.97, 94.98, 88.36, 55.79, 55.71; MS (MALDI-TOF): m/z 259 [M+Na] + ; mp 194℃
제조예 10: 에틸 5-아세트아미도벤조퓨란-2-카복실레이트Preparation 10: Ethyl 5-acetamidobenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000051
Figure PCTKR2021007223-appb-img-000051
에틸 5-아미노벤조퓨란-2-카복실레이트(205.2 mg, 1.0 mmol)를 DMF(3 mL)에 녹인 후, 여기에 (2-(1H-벤조트리아졸-1-일)-1,1,3,3-테트라메틸우로늄 헥사플루오로포스페이트(HBTU, 417.2 mg, 1.1 mmol), 히드록시벤조트리아졸(HOBt, 148.6 mg, 1.1 mmol), N,N-다이아이소프로필에틸아민(DIEA, 209.0 μL, 1.2 mmol), 및 아세트산(63.0 μL, 1.1 mmol)를 넣고 실온에서 15시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피 (CH2Cl2:EA=3:1)로 분리하여 표제 화합물 162 mg(수율 66%, 흰색 고체)을 얻었다. Ethyl 5-aminobenzofuran-2-carboxylate (205.2 mg, 1.0 mmol) was dissolved in DMF (3 mL), and then (2-(1H-benzotriazol-1-yl)-1,1,3 ,3-Tetramethyluronium hexafluorophosphate (HBTU, 417.2 mg, 1.1 mmol), hydroxybenzotriazole (HOBt, 148.6 mg, 1.1 mmol), N,N-diisopropylethylamine (DIEA, 209.0 μL) , 1.2 mmol), and acetic acid (63.0 μL, insert the 1.1 mmol) was 15 hours at room temperature. after diluting the reaction solution with water and extracted with EA., and dried the organic layer with MgSO 4, filtered, and then distilled under reduced pressure. residual Water was separated by column chromatography (CH 2 Cl 2 :EA=3:1) to obtain 162 mg (yield 66%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 10.06 (s, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 0.8 Hz, 1H), 7.65 (d, J = 8.8 Hz, 1H), 7.53 (dd, J = 8.8, 2.0 Hz, 1H), 4.35 (q, J = 7.2 Hz, 2H), 2.07 (s, 3H), 1.33 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 168.76, 159.66, 152.77, 146.68, 134.10, 127.54, 121.15, 114.18, 114.07, 112.66, 61.79, 24.64, 14.49; mp 181-182℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.06 (s, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 0.8 Hz, 1H), 7.65 (d, J) = 8.8 Hz, 1H), 7.53 (dd, J = 8.8, 2.0 Hz, 1H), 4.35 (q, J = 7.2 Hz, 2H), 2.07 (s, 3H), 1.33 (t, J = 7.2 Hz, 3H) ); 13 C NMR (100 MHz, DMSO-d 6 ): δ 168.76, 159.66, 152.77, 146.68, 134.10, 127.54, 121.15, 114.18, 114.07, 112.66, 61.79, 24.64, 14.49; mp 181-182℃
제조예 11: N-(2-(히드라진카보닐)벤조퓨란-5-일)아세트아미드Preparation 11: N-(2-(hydrazinecarbonyl)benzofuran-5-yl)acetamide
Figure PCTKR2021007223-appb-img-000052
Figure PCTKR2021007223-appb-img-000052
상기 제조예 10에서 얻은 에틸 5-아세트아미도벤조퓨란-2-카복실레이트(514.3 mg, 2.08 mmol)를 1-PrOH (20 mL)에 녹이고 여기에 히드라진 일수화물 (312.4 mg, 6.24 mmol)를 넣어 24시간 환류시켰다. 반응용액을 감압증류하고 얻어진 고체를 n-Hex과 CH2Cl2 (1:1) 혼합용매로 세척한 다음, 건조하여 표제 화합물 392 mg (81%, 흰색 고체)을 얻었다. Ethyl 5-acetamidobenzofuran-2-carboxylate (514.3 mg, 2.08 mmol) obtained in Preparation Example 10 was dissolved in 1-PrOH (20 mL), and hydrazine monohydrate (312.4 mg, 6.24 mmol) was added thereto. It was refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the resulting solid was washed with a mixed solvent of n-Hex and CH 2 Cl 2 (1:1), and then dried to obtain 392 mg (81%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 10.02 (s, 1H), 9.97 (s, 1H), 8.10 (d, J = 1.6 Hz, 1H), 7.55 (d, J = 9.2 Hz, 1H), 7.48-7.45 (m, 2H), 4.55 (br s, 2H), 2.06 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 168.23, 157.84, 150.42, 148.93, 135.37, 127.10, 119.12, 111.98, 111.72, 109.06, 23.96; MS (MALDI-TOF) m/z 256 [M+Na]+, 272 [M+K]+; mp 219-220℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.02 (s, 1H), 9.97 (s, 1H), 8.10 (d, J = 1.6 Hz, 1H), 7.55 (d, J = 9.2 Hz, 1H) ), 7.48-7.45 (m, 2H), 4.55 (br s, 2H), 2.06 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 168.23, 157.84, 150.42, 148.93, 135.37, 127.10, 119.12, 111.98, 111.72, 109.06, 23.96; MS (MALDI-TOF) m/z 256 [M+Na] + , 272 [M+K] + ; mp 219-220℃
제조예 12: 벤조[b]티오펜-2-카보히드라지드Preparation 12: benzo [b] thiophene-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000053
Figure PCTKR2021007223-appb-img-000053
벤조[b]티오펜-2-카복실산(100 mg, 0.56 mmol, Combi-blocks사)을 에탄올(2 mL)에 녹이고 진한 황산(1-2 방울)을 넣고 10시간 가열 환류하였다. 에탄올을 감압 농축시킨 다음 잔류물을 EA로 녹인후 포화 NaHCO3 수용액으로 세척하고, 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 에탄올(2 mL)에 녹이고 히드라진 일수화물(84 mg, 1.68 mmol)을 넣어 20시간 환류하였다. 반응용액을 냉각시키고 생성된 고체를 여과한 다음, 에틸 에테르로 세척하고 건조하여 표제 화합물(100 mg, 93%)을 얻었다.Benzo[b]thiophene-2-carboxylic acid (100 mg, 0.56 mmol, Combi-blocks) was dissolved in ethanol (2 mL), concentrated sulfuric acid (1-2 drops) was added, and the mixture was heated and refluxed for 10 hours. Ethanol was concentrated under reduced pressure, and the residue was dissolved in EA , washed with saturated NaHCO 3 aqueous solution, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was dissolved in ethanol (2 mL), hydrazine monohydrate (84 mg, 1.68 mmol) was added, and refluxed for 20 hours. The reaction solution was cooled and the resulting solid was filtered, washed with ethyl ether and dried to obtain the title compound (100 mg, 93%).
HRMS (TOF MS ES-): m/z calcd for C9H7N2OS(M-H)- 191.0279, found 191.0276.HRMS (TOF MS ES - ): m/z calcd for C 9 H 7 N 2 OS(MH) - 191.0279, found 191.0276.
제조예 13: 5-페닐퓨란-2-카보히드라지드Preparation 13: 5-phenylfuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000054
Figure PCTKR2021007223-appb-img-000054
5-페닐퓨란-2-카복실산(100 mg, 0.53 mmol, Combi-blocks사)을 에탄올(2 mL)에 녹이고 진한 황산(1-2 방울)을 넣고 10시간 가열 환류하였다. 에탄올을 감압 농축시킨 다음 잔류물을 EA로 녹인후 포화 NaHCO3 수용액으로 세척하고, 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 에탄올(2 mL)에 녹이고 히드라진 일수화물(84 mg, 1.68 mmol)을 넣어 20시간 환류하였다. 반응용액을 냉각시키고 생성된 고체를 여과한 다음, 에틸 에테르로 세척하고 건조하여 표제 화합물(88 mg, 82%)을 얻었다.5-phenylfuran-2-carboxylic acid (100 mg, 0.53 mmol, Combi-blocks) was dissolved in ethanol (2 mL), concentrated sulfuric acid (1-2 drops) was added, and the mixture was heated and refluxed for 10 hours. Ethanol was concentrated under reduced pressure, and the residue was dissolved in EA , washed with saturated NaHCO 3 aqueous solution, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was dissolved in ethanol (2 mL), hydrazine monohydrate (84 mg, 1.68 mmol) was added, and refluxed for 20 hours. The reaction solution was cooled and the resulting solid was filtered, washed with ethyl ether and dried to obtain the title compound (88 mg, 82%).
HRMS (TOF MS ES-): m/z calcd for C11H9N2O2(M-H)- 201.0664, found 201.0662.HRMS (TOF MS ES - ): m/z calcd for C 11 H 9 N 2 O 2 (MH) - 201.0664, found 201.0662.
제조예 14: 5-페닐티오펜-2-카보히드라지드Preparation 14: 5-phenylthiophene-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000055
Figure PCTKR2021007223-appb-img-000055
5-페닐티오펜-2-카복실산(100 mg, 0.49 mmol, Combi-blocks사)을 에탄올(2 mL)에 녹이고 진한 황산(1-2 방울)을 넣고 10시간 가열 환류하였다. 에탄올을 감압 농축시킨 다음 잔류물을 EA로 녹인후 포화 NaHCO3 수용액으로 세척하고, 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 에탄올(2 mL)에 녹이고 히드라진 일수화물(84 mg, 1.68 mmol)을 넣어 20시간 환류하였다. 반응용액을 냉각시키고 생성된 고체를 여과한 다음, 에틸 에테르로 세척하고 건조하여 표제 화합물(92 mg, 86%)을 얻었다.5-phenylthiophene-2-carboxylic acid (100 mg, 0.49 mmol, Combi-blocks) was dissolved in ethanol (2 mL), concentrated sulfuric acid (1-2 drops) was added, and the mixture was heated and refluxed for 10 hours. Ethanol was concentrated under reduced pressure, and the residue was dissolved in EA , washed with saturated NaHCO 3 aqueous solution, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was dissolved in ethanol (2 mL), hydrazine monohydrate (84 mg, 1.68 mmol) was added, and refluxed for 20 hours. The reaction solution was cooled and the resulting solid was filtered, washed with ethyl ether and dried to obtain the title compound (92 mg, 86%).
HRMS (TOF MS ES-): m/z calcd for C11H9N2OS(M-H)- 217.0436, found 217.0431.HRMS (TOF MS ES - ): m/z calcd for C 11 H 9 N 2 OS(MH) - 217.0436, found 217.0431.
제조예 15: 1-페닐-1H-피라졸-5-카보히드라지드Preparation 15: 1-phenyl-1H-pyrazole-5-carbohydrazide
Figure PCTKR2021007223-appb-img-000056
Figure PCTKR2021007223-appb-img-000056
1-페닐-1H-피라졸-5-카복실산(100 mg, 0.53 mmol, Combi-blocks사)을 에탄올(2 mL)에 녹이고 진한 황산(1-2 방울)을 넣고 10시간 가열 환류하였다. 에탄올을 감압 농축시킨 다음 잔류물을 EA로 녹인후 포화 NaHCO3 수용액으로 세척하고, 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 에탄올(2 mL)에 녹이고 히드라진 일수화물(84 mg, 1.68 mmol)을 넣어 20시간 환류하였다. 반응용액을 냉각시키고 생성된 고체를 여과한 다음, 에틸 에테르로 세척하고 건조하여 표제 화합물(91 mg, 85%)을 얻었다.1-phenyl-1H-pyrazole-5-carboxylic acid (100 mg, 0.53 mmol, Combi-blocks) was dissolved in ethanol (2 mL), concentrated sulfuric acid (1-2 drops) was added, and the mixture was heated and refluxed for 10 hours. Ethanol was concentrated under reduced pressure, and the residue was dissolved in EA , washed with saturated NaHCO 3 aqueous solution, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was dissolved in ethanol (2 mL), hydrazine monohydrate (84 mg, 1.68 mmol) was added, and refluxed for 20 hours. The reaction solution was cooled and the resulting solid was filtered, washed with ethyl ether and dried to obtain the title compound (91 mg, 85%).
HRMS (TOF MS ES-): m/z calcd for C10H9N4O(M-H)- 201.0776, found 201.0775.HRMS (TOF MS ES - ): m/z calcd for C 10 H 9 N 4 O(MH) - 201.0776, found 201.0775.
제조예 16: 에틸 나프토[2,1-b]퓨란-2-카복실레이트Preparation 16: ethyl naphtho [2,1-b] furan-2-carboxylate
Figure PCTKR2021007223-appb-img-000057
Figure PCTKR2021007223-appb-img-000057
2-히드록시-1-나프트알데히드(5.17 g, 30.0 mmol)를 DMF(80 mL)에 녹인 후, 여기에 K2CO3(20.7 g, 0.15 mol)와 에틸 브로모아세테이트(3.99 mL, 36.0 mmol)를 넣고 70℃에서 15시간 반응시켰다. 반응용액을 물로 희석한 후, EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=10:1)로 분리하여 표제 화합물 5.5 g(76%, 흰색 고체)을 얻었다. After dissolving 2-hydroxy-1-naphthaldehyde (5.17 g, 30.0 mmol) in DMF (80 mL), K 2 CO 3 (20.7 g, 0.15 mol) and ethyl bromoacetate (3.99 mL, 36.0) mmol) and reacted at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography ( n -Hex:EA=10:1) to obtain 5.5 g (76%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3) δ 8.17 (d, J = 8.0 Hz, 1H), 8.03 (s, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.88 (d, J = 9.2 Hz, 1H), 7.72 (d, J = 9.2 Hz, 1H), 7.66 (m, 1H), 7.56 (m, 1H), 4.50 (q, J = 7.2 Hz, 2H), 1.48 (t, J = 7.2 Hz, 3H); mp 94℃ 1 H NMR (400 MHz, CDCl 3 ) δ 8.17 (d, J = 8.0 Hz, 1H), 8.03 (s, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.88 (d, J = 9.2 Hz) , 1H), 7.72 (d, J = 9.2 Hz, 1H), 7.66 (m, 1H), 7.56 (m, 1H), 4.50 (q, J = 7.2 Hz, 2H), 1.48 (t, J = 7.2 Hz) , 3H); mp 94℃
제조예 17: 나프토[2,1-b]퓨란-2-카보히드라지드Preparation 17: naphtho [2,1-b] furan-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000058
Figure PCTKR2021007223-appb-img-000058
에틸 나프토[2,1-b]퓨란-2-카복실레이트(3.80 g, 15.8 mmol)를 EtOH(50 mL)에 녹이고 여기에 히드라진 일수화물(2.37 g, 47.4 mmol)를 넣어 8시간 환류시켰다. 반응용액을 냉각하고 고체를 여과하였다. 얻어진 고체를 EtOH로 세척하고 건조하여 표제 화합물 3.4 g(95%, 연노란색 고체)을 얻었다. Ethyl naphtho [2,1-b] furan-2-carboxylate (3.80 g, 15.8 mmol) was dissolved in EtOH (50 mL), hydrazine monohydrate (2.37 g, 47.4 mmol) was added thereto, and refluxed for 8 hours. The reaction solution was cooled and the solid was filtered. The obtained solid was washed with EtOH and dried to obtain 3.4 g (95%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 10.06 (s, 1H), 8.33 (d, J = 8.4 Hz, 1H), 8.18 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.96 (d, J = 9.2 Hz, 1H), 7.81 (d, J = 9.2 Hz, 1H), 7.68 (m, 1H), 7.57 (m, 1H), 4.62 (br s, 2H); 13C NMR (100 MHz, DMSO-d 6) δ 157.90, 152.03, 147.96, 130.04, 128.81, 127.77, 127.45, 127.15, 125.28, 123.59, 122.55, 112.51, 108.37; MS (MALDI-TOF) m/z 249 [M+Na]+; mp 248℃ 1 H NMR (400 MHz, DMSO- d 6 ) δ 10.06 (s, 1H), 8.33 (d, J = 8.4 Hz, 1H), 8.18 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H) , 7.96 (d, J = 9.2 Hz, 1H), 7.81 (d, J = 9.2 Hz, 1H), 7.68 (m, 1H), 7.57 (m, 1H), 4.62 (br s, 2H); 13 C NMR (100 MHz, DMSO- d 6 ) δ 157.90, 152.03, 147.96, 130.04, 128.81, 127.77, 127.45, 127.15, 125.28, 123.59, 122.55, 112.51, 108.37; MS (MALDI-TOF) m/z 249 [M+Na] + ; mp 248℃
<방향족/N-치환된 인돌-3-카복스알데히드 유도체의 제조><Preparation of aromatic/N-substituted indole-3-carboxaldehyde derivatives>
Figure PCTKR2021007223-appb-img-000059
Figure PCTKR2021007223-appb-img-000059
제조예 18: 2-클로로-1-메틸-1H-인돌-3-카복스알데히드Preparation 18: 2-chloro-1-methyl-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000060
Figure PCTKR2021007223-appb-img-000060
0℃에서 2-클로로-1H-인돌-3-카복스알데히드(359.2 mg, 2.0 mmol)와 NaH(120.0 mg, 3.0 mmol, 오일 중 60%)에 THF(20 mL)를 넣고 5분 교반한 후, 아이오도메탄(149.4 μL, 2.4 mmol)를 넣고 실온에서 5시간 교반하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=15:1)로 분리하여 표제 화합물 330 mg(수율 85%, 흰색 고체)을 얻었다. THF (20 mL) was added to 2-chloro-1H-indole-3-carboxaldehyde (359.2 mg, 2.0 mmol) and NaH (120.0 mg, 3.0 mmol, 60% in oil) at 0° C. and stirred for 5 minutes. , iodomethane (149.4 μL, 2.4 mmol) was added and stirred at room temperature for 5 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH 2 Cl 2 :EA=15:1) to obtain 330 mg (yield 85%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.13 (s, 1H), 8.30 (m, 1H), 7.36-7.32 (m, 3H), 3.82 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 184.07, 136.99, 136.13, 124.44, 124.20, 123.69, 121.43, 113.02, 109.64, 30.28; MS (MALDI-TOF): m/z 194 [M+H]+; mp 97-98℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.13 (s, 1H), 8.30 (m, 1H), 7.36-7.32 (m, 3H), 3.82 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 184.07, 136.99, 136.13, 124.44, 124.20, 123.69, 121.43, 113.02, 109.64, 30.28; MS (MALDI-TOF): m/z 194 [M+H] + ; mp 97-98℃
제조예 19: 에틸 2-(2-클로로-3-포밀-1H-인돌-1-일)아세테이트Preparation 19: ethyl 2- (2-chloro-3-formyl-1H-indol-1-yl) acetate
Figure PCTKR2021007223-appb-img-000061
Figure PCTKR2021007223-appb-img-000061
0℃에서 2-클로로-1H-인돌-3-카복스알데히드(200.0 mg, 1.11 mmol)와 NaH(66.8 mg, 1.67 mmol, 오일 중 60%)에 THF (10 mL)를 넣고 5분 교반한 후, 에틸 브로모아세테이트(147.5 μL, 1.33 mmol)를 넣고 실온에서 7시간 교반하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=20:1)로 분리하여 표제 화합물 244 mg(수율 83%, 흰색 고체)을 얻었다. THF (10 mL) was added to 2-chloro-1H-indole-3-carboxaldehyde (200.0 mg, 1.11 mmol) and NaH (66.8 mg, 1.67 mmol, 60% in oil) at 0° C. and stirred for 5 minutes. , ethyl bromoacetate (147.5 μL, 1.33 mmol) was added, and the mixture was stirred at room temperature for 7 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH 2 Cl 2 :EA=20:1) to obtain 244 mg (yield 83%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.16 (s, 1H), 8.32 (m, 1H), 7.36-7.32 (m, 2H), 7.23 (m, 1H), 4.95 (s, 2H), 4.26 (q, J = 7.2 Hz, 2H), 1.28 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 184.09, 166.47, 136.48, 135.70, 124.45, 124.34, 123.78, 121.55, 113.62, 109.03, 62.36, 44.81, 14.05; MS (MALDI-TOF): m/z 266 [M+H]+; mp 110℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.16 (s, 1H), 8.32 (m, 1H), 7.36-7.32 (m, 2H), 7.23 (m, 1H), 4.95 (s, 2H), 4.26 (q, J = 7.2 Hz, 2H), 1.28 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 184.09, 166.47, 136.48, 135.70, 124.45, 124.34, 123.78, 121.55, 113.62, 109.03, 62.36, 44.81, 14.05; MS (MALDI-TOF): m/z 266 [M+H] + ; mp 110℃
제조예 20: 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드Preparation 20: 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000062
Figure PCTKR2021007223-appb-img-000062
2-클로로-1H-인돌-3-카복스알데히드(2.69 g, 15.0 mmol)를 DMF(50 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(2.01 mL, 18.0 mmol)와 Cs2CO3(14.7 g, 45.0 mmol)를 넣고 70℃에서 15시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:CH2Cl2:EA=4:2:1)로 분리하여 표제 화합물 2.95 g(수율 78%, 연노란색 고체)을 얻었다. After dissolving 2-chloro-1H-indole-3-carboxaldehyde (2.69 g, 15.0 mmol) in DMF (50 mL), 2-bromoethyl ethyl ether (2.01 mL, 18.0 mmol) and Cs 2 CO 3 ( 14.7 g, 45.0 mmol) was added and heated at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:CH 2 Cl 2 :EA=4:2:1) to obtain 2.95 g (yield 78%, pale yellow solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.14 (s, 1H), 8.30 (m, 1H), 7.41 (m, 1H), 7.34-7.30 (m, 2H), 4.42 (t, J = 5.6 Hz, 2H), 3.76 (t, J = 5.6 Hz, 2H), 3.44 (q, J = 7.2 Hz, 2H), 1.12 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 184.09, 136.58, 135.80, 124.41, 123.95, 123.43, 121.24, 113.08, 110.04, 68.13, 66.91, 43.97, 14.97; MS (MALDI-TOF): m/z 252 [M+H]+; mp 52℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.14 (s, 1H), 8.30 (m, 1H), 7.41 (m, 1H), 7.34-7.30 (m, 2H), 4.42 (t, J = 5.6 Hz) , 2H), 3.76 (t, J = 5.6 Hz, 2H), 3.44 (q, J = 7.2 Hz, 2H), 1.12 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 184.09, 136.58, 135.80, 124.41, 123.95, 123.43, 121.24, 113.08, 110.04, 68.13, 66.91, 43.97, 14.97; MS (MALDI-TOF): m/z 252 [M+H] + ; mp 52℃
제조예 21: 2-클로로-1-(2-메톡시에틸)-1H-인돌-3-카복스알데히드Preparation 21: 2-chloro-1-(2-methoxyethyl)-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000063
Figure PCTKR2021007223-appb-img-000063
2-클로로-1H-인돌-3-카복스알데히드(538.8 mg, 3.0 mmol)를 DMF(10 mL)에 녹인 후, 2-브로모에틸 메틸 에테르(422.6 μL, 4.5 mmol)와 Cs2CO3(2.93 g, 9.0 mmol)를 넣고 70℃에서 15시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=2:1)로 분리하여 표제 화합물 468 mg(수율 66%, 연노란색 고체)을 얻었다. After dissolving 2-chloro-1H-indole-3-carboxaldehyde (538.8 mg, 3.0 mmol) in DMF (10 mL), 2-bromoethyl methyl ether (422.6 μL, 4.5 mmol) and Cs 2 CO 3 ( 2.93 g, 9.0 mmol) and heated at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=2:1) to obtain 468 mg (yield 66%, pale yellow solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.14 (s, 1H), 8.30 (m, 1H), 7.42-7.31 (m, 3H), 4.43 (t, J = 5.7 Hz, 2H), 3.74 (t, J = 5.7 Hz, 2H), 3.32 (s, 3H); 13C NMR (100 MHz, CDCl3): δ 184.27, 136.79, 135.97, 124.56, 124.21, 123.64, 121.44, 113.25, 110.17, 70.49, 59.35, 44.00; MS (MALDI-TOF): m/z 238 [M+H]+, 260 [M+Na]+; mp 66-67℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.14 (s, 1H), 8.30 (m, 1H), 7.42-7.31 (m, 3H), 4.43 (t, J = 5.7 Hz, 2H), 3.74 (t) , J = 5.7 Hz, 2H), 3.32 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 184.27, 136.79, 135.97, 124.56, 124.21, 123.64, 121.44, 113.25, 110.17, 70.49, 59.35, 44.00; MS (MALDI-TOF): m/z 238 [M+H] + , 260 [M+Na] + ; mp 66-67℃
제조예 22: 5-메틸옥시인돌Preparation 22: 5-methyloxyindole
Figure PCTKR2021007223-appb-img-000064
Figure PCTKR2021007223-appb-img-000064
5-메틸이사틴(1.50 g, 9.31 mmol)에 에틸렌 글리콜(10 mL)을 넣은 후, KOH(522.4 mg, 9.31 mmol)와 히드라진 일수화물(1.40 g, 27.9 mmol)를 넣고 140℃에서 4시간 가열하였다. 반응용액을 냉각하고 1 N HCl로 산성화한 후, EA로 추출하였다. 유기층을 감압 증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=3:2)로 분리하여 표제 화합물 1.0 g(수율 73%, 연갈색 고체)을 얻었다. After adding ethylene glycol (10 mL) to 5-methylisatin (1.50 g, 9.31 mmol), KOH (522.4 mg, 9.31 mmol) and hydrazine monohydrate (1.40 g, 27.9 mmol) were added and heated at 140°C for 4 hours. did The reaction solution was cooled, acidified with 1 N HCl, and extracted with EA. The organic layer was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=3:2) to obtain 1.0 g of the title compound (yield 73%, light brown solid).
1H NMR (400 MHz, DMSO-d6): δ 10.23 (br s, 1H), 7.01 (s, 1H), 6.96 (d, J = 7.9 Hz, 1H), 6.69 (d, J = 7.9 Hz, 1H), 3.41 (s, 2H), 2.23 (s, 3H) 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.23 (br s, 1H), 7.01 (s, 1H), 6.96 (d, J = 7.9 Hz, 1H), 6.69 (d, J = 7.9 Hz, 1H), 3.41 (s, 2H), 2.23 (s, 3H)
제조예 23: 2-클로로-5-메틸-1H-인돌-3-카복스알데히드Preparation 23: 2-chloro-5-methyl-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000065
Figure PCTKR2021007223-appb-img-000065
0℃에서 DMF(10 mL)에 POCl3(2.65 mL, 28.9 mmol)를 넣고 10분 교반한 후, 5-메틸옥시인돌(850.0 mg, 5.78 mmol)의 DMF(10 mL) 용액을 넣고 80℃에서 3시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, EA로 추출하였다. 유기층을 물로 세척하고 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA= 3:2)로 분리하여 표제 화합물 600 mg(수율 54%, 연노란색 고체)을 얻었다. POCl 3 (2.65 mL, 28.9 mmol) was added to DMF (10 mL) at 0° C., and stirred for 10 minutes. Then, a DMF (10 mL) solution of 5-methyloxyindole (850.0 mg, 5.78 mmol) was added and at 80° C. Heated for 3 hours. The reaction solution was alkalized by adding 1 N NaOH, followed by extraction with EA. The organic layer was washed with water, dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA = 3:2) to obtain 600 mg of the title compound (yield 54%, pale yellow solid).
1H NMR (400 MHz, DMSO-d6): δ 12.96 (s, 1H), 9.96 (s, 1H), 7.87(s, 1H), 7.31 (d, J = 8.3 Hz), 7.10 (dd, J = 8.3, 1.5 Hz, 1H), 2.39 (s, 3H); mp 215℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.96 (s, 1H), 9.96 (s, 1H), 7.87(s, 1H), 7.31 (d, J = 8.3 Hz), 7.10 (dd, J) = 8.3, 1.5 Hz, 1H), 2.39 (s, 3H); mp 215℃
제조예 24: 2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-카복스알데히드Preparation 24: 2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000066
Figure PCTKR2021007223-appb-img-000066
2-클로로-5-메틸-1H-인돌-3-카복스알데히드(440.0 mg, 2.27 mmol)의 CH3CN(20 mL) 용액에 2-브로모에틸 에틸 에테르(303.8 μL, 2.72 mmol)와 Cs2CO3(3.71 g, 11.4 mmol)를 넣고 15시간 환류하였다. 반응용액을 감압 증류하고 잔류물에 물을 넣어 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=9:1)로 분리하여 표제 화합물 570 mg (94%, 흰색 고체)을 얻었다.In a solution of 2-chloro-5-methyl-1H-indole-3-carboxaldehyde (440.0 mg, 2.27 mmol) in CH 3 CN (20 mL), 2-bromoethyl ethyl ether (303.8 μL, 2.72 mmol) and Cs 2 CO 3 (3.71 g, 11.4 mmol) was added and refluxed for 15 hours. The reaction solution was distilled under reduced pressure, and water was added to the residue, followed by extraction with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=9:1) to give 570 mg (94%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.09 (s, 1H), 8.10 (s, 1H), 7.28 (d, J = 8.5 Hz, 1H), 7.14 (d, J = 8.5, 1.5 Hz, 1H), 4.39 (t, J = 5.5 Hz, 2H), 3.74 (t, J = 5.5 Hz, 2H), 3.43 (q, J = 7.0 Hz, 2H), 2.46 (s, 3H), 1.11 (t, J = 7.0 Hz, 3H); 13C NMR (400 MHz, DMSO-d6): δ 184.35, 136.69, 134.28, 133.48, 125.59, 124.73, 121.19, 112.92, 109.89, 68.30, 67.09, 44.19, 21.61, 15.17; MS (MALDI-TOF): m/z 266 [M+H]+; mp 54℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.09 (s, 1H), 8.10 (s, 1H), 7.28 (d, J = 8.5 Hz, 1H), 7.14 (d, J = 8.5, 1.5 Hz, 1H) ), 4.39 (t, J = 5.5 Hz, 2H), 3.74 (t, J = 5.5 Hz, 2H), 3.43 (q, J = 7.0 Hz, 2H), 2.46 (s, 3H), 1.11 (t, J = 7.0 Hz, 3H); 13 C NMR (400 MHz, DMSO-d 6 ): δ 184.35, 136.69, 134.28, 133.48, 125.59, 124.73, 121.19, 112.92, 109.89, 68.30, 67.09, 44.19, 21.61, 15.17; MS (MALDI-TOF): m/z 266 [M+H] + ; mp 54℃
제조예 25: 2-클로로-5-메톡시-1H-인돌-3-카복스알데히드Preparation 25: 2-chloro-5-methoxy-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000067
Figure PCTKR2021007223-appb-img-000067
0℃에서 DMF(10 mL)에 POCl3(2.24 mL g, 24.5 mmol)를 넣고 10분 교반한 후, 5-메톡시옥시인돌(1.60 g, 9.81 mml)의 DMF(10 mL) 용액을 넣고 80℃에서 2시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, 생성된 고체는 물로 세척하고 건조하여 표제 화합물을 얻었다. 여액은 다시 EA로 추출하고 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:2)로 분리하여 표제 화합물 1.27 g(수율 62%, 연갈색 고체)을 얻었다. POCl 3 (2.24 mL g, 24.5 mmol) was added to DMF (10 mL) at 0° C. and stirred for 10 minutes. Then, a DMF (10 mL) solution of 5-methoxyoxyindole (1.60 g, 9.81 mml) was added and 80 It was heated at ℃ for 2 hours. After alkalizing the reaction solution by adding 1 N NaOH, the resulting solid was washed with water and dried to obtain the title compound. The filtrate was again extracted with EA, dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=1:2) to obtain 1.27 g (yield 62%, light brown solid) of the title compound.
1H NMR (300 MHz, DMSO-d6): δ 12.96 (br s, 1H), 9.96 (s, 1H), 7.57 (d, J = 2.7 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.90 (dd, J = 8.7, 2.7 Hz, 1H), 3.79 (s, 3H); mp 225℃ 1 H NMR (300 MHz, DMSO-d 6 ): δ 12.96 (br s, 1H), 9.96 (s, 1H), 7.57 (d, J = 2.7 Hz, 1H), 7.33 (d, J = 8.7 Hz, 1H), 6.90 (dd, J = 8.7, 2.7 Hz, 1H), 3.79 (s, 3H); mp 225℃
제조예 26: 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드Preparation 26: 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000068
Figure PCTKR2021007223-appb-img-000068
5-메톡시-2-클로로-1H-인돌-3-카복스알데히드(1.97 g, 9.40 mmol)를 DMF(70 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(1.26 mL, 11.3 mmol)와 Cs2CO3(15.3 g, 47.0 mmol)를 넣고 70℃에서 11시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=3:1)로 분리하여 표제 화합물 2.18 g(수율 82%, 연노란색 고체)을 얻었다.5-Methoxy-2-chloro-1H-indole-3-carboxaldehyde (1.97 g, 9.40 mmol) was dissolved in DMF (70 mL), followed by 2-bromoethyl ethyl ether (1.26 mL, 11.3 mmol) and Cs 2 CO 3 (15.3 g, 47.0 mmol) was added and heated at 70° C. for 11 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=3:1) to obtain 2.18 g (yield 82%, pale yellow solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.09 (s, 1H), 7.79 (d, J = 2.7 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H), 6.94 (dd, J = 9.0, 2.7 Hz, 1H), 4.37 (t, J = 5.7 Hz, 2H), 3.89 (s, 3H), 3.74 (t, J = 5.7 Hz, 2H), 3.43 (q, J = 7.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 184.10, 156.90, 135.99, 130.59, 125.12, 114.17, 112.95, 111.06, 102.64, 68.21, 66.90, 55.77, 44.16, 14.96; MS (MALDI-TOF): m/z 282 [M+H]+; mp 48℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.09 (s, 1H), 7.79 (d, J = 2.7 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H), 6.94 (dd, J = 9.0) , 2.7 Hz, 1H), 4.37 (t, J = 5.7 Hz, 2H), 3.89 (s, 3H), 3.74 (t, J = 5.7 Hz, 2H), 3.43 (q, J = 7.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 184.10, 156.90, 135.99, 130.59, 125.12, 114.17, 112.95, 111.06, 102.64, 68.21, 66.90, 55.77, 44.16, 14.96; MS (MALDI-TOF): m/z 282 [M+H] + ; mp 48℃
제조예 27: 6-메톡시옥시인돌Preparation 27: 6-methoxyoxyindole
Figure PCTKR2021007223-appb-img-000069
Figure PCTKR2021007223-appb-img-000069
6-메톡시이사틴(500 mg, 2.82 mmol)에 에틸렌 글리콜(10 mL)을 넣은 후, KOH(158.2 mg, 2.82 mmol)와 히드라진 일수화물(282.3 mg, 5.64 mmol)를 넣고 140℃에서 4시간 가열하였다. 반응용액을 냉각하고 1 N HCl로 산성화한 후, EA로 추출하였다. 유기층을 감압 증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 278 mg(수율 60%, 연노란색 고체)을 얻었다. After adding ethylene glycol (10 mL) to 6-methoxyisatin (500 mg, 2.82 mmol), KOH (158.2 mg, 2.82 mmol) and hydrazine monohydrate (282.3 mg, 5.64 mmol) were added and heated at 140°C for 4 hours. did The reaction solution was cooled, acidified with 1 N HCl, and extracted with EA. The organic layer was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=1:1) to obtain 278 mg (yield: 60%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 10.30 (br s, 1H), 7.08 (d, J = 8.0 Hz, 1H), 6.48 (d, J = 8.0, 2.4 Hz, 1H), 6.38 (d, J = 2.4 Hz, 1H), 3.71 (s, 3H), 3.37 (s, 2H) 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.30 (br s, 1H), 7.08 (d, J = 8.0 Hz, 1H), 6.48 (d, J = 8.0, 2.4 Hz, 1H), 6.38 ( d, J = 2.4 Hz, 1H), 3.71 (s, 3H), 3.37 (s, 2H)
제조예 28: 2-클로로-6-메톡시-1H-인돌-3-카복스알데히드Preparation 28: 2-chloro-6-methoxy-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000070
Figure PCTKR2021007223-appb-img-000070
0℃에서 DMF(5 mL)에 POCl3(1.79 mL g, 19.5 mmol)를 넣고 10분 교반한 후, 6-메톡시옥시인돌(1.27 g, 7.78 mmol)의 DMF(15 mL) 용액을 넣고 80℃에서 2시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, 생성된 고체는 여과하여 물로 세척하고 건조하여 표제 화합물을 얻었다. 여액은 다시 EA로 추출하고 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=10:1)로 분리하여 표제 화합물 945 mg(수율 흰색 고체)을 얻었다. POCl 3 (1.79 mL g, 19.5 mmol) was added to DMF (5 mL) at 0° C., stirred for 10 minutes, and a DMF (15 mL) solution of 6-methoxyoxyindole (1.27 g, 7.78 mmol) was added, and 80 It was heated at ℃ for 2 hours. After alkalizing the reaction solution by adding 1 N NaOH, the resulting solid was filtered, washed with water, and dried to obtain the title compound. The filtrate was again extracted with EA, dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH 2 Cl 2 :EA=10:1) to obtain 945 mg (yield white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.87 (br s, 1H), 9.94 (s, 1H), 7.91 (m, 1H), 6.89-6.87 (m, 2H), 3.79 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 183.09, 156.90, 135.57, 133.26, 120.73, 118.09, 112.23, 112.10, 95.09, 55.31; mp 230℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.87 (br s, 1H), 9.94 (s, 1H), 7.91 (m, 1H), 6.89-6.87 (m, 2H), 3.79 (s, 3H) ); 13 C NMR (100 MHz, DMSO-d 6 ): δ 183.09, 156.90, 135.57, 133.26, 120.73, 118.09, 112.23, 112.10, 95.09, 55.31; mp 230℃
제조예 29: 2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-카복스알데히드Preparation 29: 2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000071
Figure PCTKR2021007223-appb-img-000071
6-메톡시-2-클로로-1H-인돌-3-카복스알데히드(765.1 mg, 3.65 mmol)를 DMF(15 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(489.2 μL, 4.38 mmol)와 Cs2CO3(3.58 g, 11.0 mmol)를 넣고 70℃에서 5시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:CH2Cl2:EA=3:1:0.5)로 분리하여 표제 화합물 420 mg(수율 41%, 흰색 고체)을 얻었다. 6-Methoxy-2-chloro-1H-indole-3-carboxaldehyde (765.1 mg, 3.65 mmol) was dissolved in DMF (15 mL), followed by 2-bromoethyl ethyl ether (489.2 μL, 4.38 mmol) and Cs 2 CO 3 (3.58 g, 11.0 mmol) was added and heated at 70° C. for 5 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:CH 2 Cl 2 :EA=3:1:0.5) to obtain 420 mg (yield 41%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.08 (s, 1H), 8.15 (d, J = 8.8 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H), 6.89 (d, J = 2.4 Hz, 1H), 4.36 (t, J = 5.6 Hz, 2H), 3.87 (s, 3H), 3.75 (t, J = 5.6 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.13 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 184.02, 157.51, 136.85, 135.25, 122.00, 118.29, 113.20, 112.31, 94.46, 68.25, 66.92, 55.68, 44.00, 15.04; MS (MALDI-TOF): m/z 281 [M]+; mp 82℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.08 (s, 1H), 8.15 (d, J = 8.8 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H), 6.89 (d, J) = 2.4 Hz, 1H), 4.36 (t, J = 5.6 Hz, 2H), 3.87 (s, 3H), 3.75 (t, J = 5.6 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.13 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 184.02, 157.51, 136.85, 135.25, 122.00, 118.29, 113.20, 112.31, 94.46, 68.25, 66.92, 55.68, 44.00, 15.04; MS (MALDI-TOF): m/z 281 [M] + ; mp 82℃
제조예 30: 2-클로로-5-플루오로-1H-인돌-3-카복스알데히드Preparation 30: 2-chloro-5-fluoro-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000072
Figure PCTKR2021007223-appb-img-000072
0℃에서 DMF(5 mL)에 POCl3(1.36 mL g, 14.9 mmol)를 넣고 10분 교반한 후, 5-플루오로옥시인돌(900.0 mg, 5.95 mmol)의 DMF(15 mL) 용액을 넣고 80℃에서 4시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, EA로 추출하였다. 유기층을 Na2SO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 235 mg(수율 20%, 흰색 고체)을 얻었다. POCl 3 (1.36 mL g, 14.9 mmol) was added to DMF (5 mL) at 0° C., and stirred for 10 minutes. Then, a DMF (15 mL) solution of 5-fluorooxyindole (900.0 mg, 5.95 mmol) was added and 80 It was heated at ℃ for 4 hours. The reaction solution was alkalized by adding 1 N NaOH, followed by extraction with EA. The organic layer was dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=1:1) to obtain 235 mg (yield 20%, white solid) of the title compound.
1H NMR (300 MHz, DMSO-d6): δ 13.22 (br s, 1H), 9.96 (s, 1H), 7.73 (dd, J = 9.0, 2.7 Hz, 1H), 7.45 (dd, J = 9.0, 4.5 Hz, 1H), 7.13 (m, 1H); 13C NMR (100 MHz, DMSO-d6): δ 183.27, 158.93 (d, J = 235.2 Hz), 135.60, 131.21, 124.89 (d, J = 11.3 Hz), 113.27 (d, J = 9.9 Hz), 112.11 (d, J = 4.4 Hz), 111.92 (d, J = 25.9 Hz), 105.09 (d, J = 25.1 Hz); mp 208-210℃ 1 H NMR (300 MHz, DMSO-d 6 ): δ 13.22 (br s, 1H), 9.96 (s, 1H), 7.73 (dd, J = 9.0, 2.7 Hz, 1H), 7.45 (dd, J = 9.0) , 4.5 Hz, 1H), 7.13 (m, 1H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 183.27, 158.93 (d, J = 235.2 Hz), 135.60, 131.21, 124.89 (d, J = 11.3 Hz), 113.27 (d, J = 9.9 Hz), 112.11 (d, J = 4.4 Hz), 111.92 (d, J = 25.9 Hz), 105.09 (d, J = 25.1 Hz); mp 208-210℃
제조예 31: 2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-카복스알데히드Preparation 31: 2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000073
Figure PCTKR2021007223-appb-img-000073
2-클로로-5-플루오로-1H-인돌-3-카복스알데히드(197.6 mg, 1.0 mmol)를 DMF(5 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(134.0 μL, 1.2 mmol)와 Cs2CO3(977.5 mg, 3.0 mmol)를 넣고 70℃에서 15시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:CH2Cl2:EA=3:1:0.5)로 분리하여 표제 화합물 125 mg(수율 46%, 흰색 고체)을 얻었다.2-Chloro-5-fluoro-1H-indole-3-carboxaldehyde (197.6 mg, 1.0 mmol) was dissolved in DMF (5 mL), followed by 2-bromoethyl ethyl ether (134.0 μL, 1.2 mmol) and Cs 2 CO 3 (977.5 mg, 3.0 mmol) was added and heated at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:CH 2 Cl 2 :EA=3:1:0.5) to obtain 125 mg (yield 46%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.09 (s, 1H), 7.98 (d, J = 9.2, 2.4 Hz, 1H), 7.36 (d, J = 9.2, 4.4 Hz, 1H), 7.06 (m, 1H), 4.41 (t, J = 5.6 Hz, 2H), 3.76 (t, J = 5.6 Hz, 2H), 3.43 (q, J = 7.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H); 13C NMR (400 MHz, DMSO-d6): δ 183.79, 159.96 (d, J = 238.6 Hz), 137.13, 132.38, 124.95 (d, J = 11.3 Hz), 113.07 (d, J = 4.4 Hz), 112.19 (d, J = 26.2 Hz), 111.30 (d, J = 9.4 Hz), 106.80 (d, J = 25.1 Hz), 68.23, 66.91, 44.37, 14.93; MS (MALDI-TOF): m/z 270 [M+H]+; mp 83-84℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.09 (s, 1H), 7.98 (d, J = 9.2, 2.4 Hz, 1H), 7.36 (d, J = 9.2, 4.4 Hz, 1H), 7.06 (m , 1H), 4.41 (t, J = 5.6 Hz, 2H), 3.76 (t, J = 5.6 Hz, 2H), 3.43 (q, J = 7.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H) ); 13 C NMR (400 MHz, DMSO-d 6 ): δ 183.79, 159.96 (d, J = 238.6 Hz), 137.13, 132.38, 124.95 (d, J = 11.3 Hz), 113.07 (d, J = 4.4 Hz), 112.19 (d, J = 26.2 Hz), 111.30 (d, J = 9.4 Hz), 106.80 (d, J = 25.1 Hz), 68.23, 66.91, 44.37, 14.93; MS (MALDI-TOF): m/z 270 [M+H] + ; mp 83-84℃
제조예 32: 2,5-디클로로-1H-인돌-3-카복스알데히드Preparation 32: 2,5-dichloro-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000074
Figure PCTKR2021007223-appb-img-000074
0℃에서 DMF(5 mL)에 POCl3(2.74 mL, 29.9 mmol)를 넣고 10분 교반한 후, 5-클로로옥시인돌(1.0 g, 5.97 mmol)의 DMF(5 mL) 용액을 넣고 80℃에서 3시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, EA로 추출하였다. 유기층을 물로 세척하고 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=2:1)로 분리하여 표제 화합물 530 mg(수율 41%, 연갈색 고체)을 얻었다. POCl 3 (2.74 mL, 29.9 mmol) was added to DMF (5 mL) at 0° C. and stirred for 10 minutes. Then, a DMF (5 mL) solution of 5-chlorooxyindole (1.0 g, 5.97 mmol) was added and at 80° C. Heated for 3 hours. The reaction solution was alkalized by adding 1 N NaOH, followed by extraction with EA. The organic layer was washed with water, dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=2:1) to obtain 530 mg (yield 41%, light brown solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 13.31 (br s, 1H), 9.97 (s, 1H), 8.04 (d, J = 2.0 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.31 (dd, J = 8.5, 2.0 Hz, 1H); mp 245-248℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.31 (br s, 1H), 9.97 (s, 1H), 8.04 (d, J = 2.0 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.31 (dd, J = 8.5, 2.0 Hz, 1H); mp 245-248℃
제조예 33: 2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드Preparation 33: 2,5-dichloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000075
Figure PCTKR2021007223-appb-img-000075
상기 제조예 32에서 얻은 2,5-디클로로-1H-인돌-3-카복스알데히드(350 mg, 1.64 mmol)의 CH3CN(20 mL) 용액에 2-브로모에틸 에틸 에테르(274.8 μL, 2.46 mmol)와 Cs2CO3(2.67 g, 8.20 mmol)를 넣고 15시간 환류하였다. 반응용액을 감압 증류하고 잔류물에 물을 넣어 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=9:1)로 분리하여 표제 화합물 360 mg(수율 77%, 연노란색 고체)을 얻었다.2-bromoethyl ethyl ether (274.8 μL, 2.46 ) in CH 3 CN (20 mL) solution of 2,5-dichloro-1H-indole-3-carboxaldehyde (350 mg, 1.64 mmol) obtained in Preparation 32 above mmol) and Cs 2 CO 3 (2.67 g, 8.20 mmol) were added and refluxed for 15 hours. The reaction solution was distilled under reduced pressure, and water was added to the residue, followed by extraction with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=9:1) to obtain 360 mg (yield 77%, pale yellow solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.06 (s, 1H), 8.26 (d, J = 2.0 Hz, 1H), 7.32-7.23 (m, 2H), 4.38 (t, J = 5.6 Hz, 2H), 3.73 (t, J = 5.6 Hz, 2H), 3.41 (q, J = 7.2 Hz, 2H), 1.08 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 184.01, 137.39, 134.50, 129.64, 125.40, 124.51, 120.95, 112.84, 111.60, 68.41, 67.13, 44.55, 15.15; MS (MALDI-TOF): m/z 286 [M+H]+; mp 104℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.06 (s, 1H), 8.26 (d, J = 2.0 Hz, 1H), 7.32-7.23 (m, 2H), 4.38 (t, J = 5.6 Hz, 2H) ), 3.73 (t, J = 5.6 Hz, 2H), 3.41 (q, J = 7.2 Hz, 2H), 1.08 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 184.01, 137.39, 134.50, 129.64, 125.40, 124.51, 120.95, 112.84, 111.60, 68.41, 67.13, 44.55, 15.15; MS (MALDI-TOF): m/z 286 [M+H] + ; mp 104℃
제조예 34: 5-(트리플루오로메톡시)옥시인돌Preparation 34: 5- (trifluoromethoxy) oxyindole
Figure PCTKR2021007223-appb-img-000076
Figure PCTKR2021007223-appb-img-000076
5-(트리플루오로메톡시)이사틴(1.75 g, 7.57 mmol)에 에틸렌 글리콜(10 mL)을 넣은 후, KOH(424.8 mg, 7.57 mmol)와 히드라진 일수화물(1.14 g, 22.7 mmol)를 넣고 140℃에서 4시간 가열하였다. 반응용액을 냉각하고 1 N HCl로 산성화한 후, EA로 추출하였다. 유기층을 감압 증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:2)로 분리하여 표제 화합물 855 mg(52%, 연갈색 고체)을 얻었다. After adding ethylene glycol (10 mL) to 5-(trifluoromethoxy) isatin (1.75 g, 7.57 mmol), KOH (424.8 mg, 7.57 mmol) and hydrazine monohydrate (1.14 g, 22.7 mmol) were added 140 It was heated at ℃ for 4 hours. The reaction solution was cooled, acidified with 1 N HCl, and extracted with EA. The organic layer was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=1:2) to obtain 855 mg (52%, light brown solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 10.52 (br s, 1H), 7.24 (s, 1H), 7.16 (m, 1H), 6.86 (d, J = 8.0 Hz, 1H), 3.54 (s, 2H) 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.52 (br s, 1H), 7.24 (s, 1H), 7.16 (m, 1H), 6.86 (d, J = 8.0 Hz, 1H), 3.54 ( s, 2H)
제조예 35: 2-클로로-5-(트리플루오로메톡시)-1H-인돌-3-카복스알데히드Preparation 35: 2-chloro-5- (trifluoromethoxy) -1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000077
Figure PCTKR2021007223-appb-img-000077
0℃에서 DMF(1 mL)에 POCl3(727.7 μL g, 7.95 mmol)를 넣고 10분 교반한 후, 5-(트리플루오로메톡시)옥시인돌(575.0 mg, 2.65 mmol)의 DMF(5 mL) 용액을 넣고 80℃에서 3시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, EA로 추출하였다. 유기층을 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 120 mg(수율 17%, 흰색 고체)을 얻었다. POCl 3 (727.7 μL g, 7.95 mmol) was added to DMF (1 mL) at 0° C. and stirred for 10 minutes, followed by 5-(trifluoromethoxy) oxyindole (575.0 mg, 2.65 mmol) in DMF (5 mL) The solution was added and heated at 80° C. for 3 hours. The reaction solution was alkalized by adding 1 N NaOH, followed by extraction with EA. The organic layer was dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=1:1) to obtain 120 mg (yield 17%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 13.40 (br s, 1H), 9.99 (s, 1H), 7.95 (m, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.29 (m, 1H); mp 191-192℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.40 (br s, 1H), 9.99 (s, 1H), 7.95 (m, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.29 ( m, 1H); mp 191-192℃
제조예 36: 2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-카복스알데히드Preparation 36: 2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000078
Figure PCTKR2021007223-appb-img-000078
2-클로로-5-(트리플루오로메톡시)-1H-인돌-3-카복스알데히드(100 mg, 0.38 mmol)를 DMF (3 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(51.4 μL, 0.46 mmol)와 Cs2CO3(371.4 mg, 1.14 mmol)를 넣고 70℃에서 8시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:CH2Cl2:EA =4:4:1)로 분리하여 표제 화합물 43 mg(수율 34%, 연노란색 고체)을 얻었다.After dissolving 2-chloro-5- (trifluoromethoxy) -1H-indole-3-carboxaldehyde (100 mg, 0.38 mmol) in DMF (3 mL), 2-bromoethyl ethyl ether (51.4 μL, 0.46 mmol) and Cs 2 CO 3 (371.4 mg, 1.14 mmol) were added and heated at 70° C. for 8 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:CH 2 Cl 2 :EA =4:4:1) to obtain 43 mg of the title compound (yield 34%, pale yellow solid).
1H NMR (400 MHz, CDCl3): δ 10.11 (s, 1H), 8.18 (s, 1H), 7.42 (d, J = 8.8 Hz, 1H), 7.20 (dd, J = 8.8, 1.6 Hz, 1H), 4.42 (t, J = 5.6 Hz, 2H), 3.77 (t, J = 5.6 Hz, 2H), 3.44 (q, J = 5.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 183.78, 145.66 (q, J = 1.9 Hz), 137.58, 134.20, 124.73, 120.62 (q, J = 255.0 Hz), 117.81, 113.86 (q, J = 0.8 Hz), 113.23, 111.29, 68.25, 66.96, 44.46, 14.94; MS (MALDI-TOF): m/z 335 [M]+; mp 79℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.11 (s, 1H), 8.18 (s, 1H), 7.42 (d, J = 8.8 Hz, 1H), 7.20 (dd, J = 8.8, 1.6 Hz, 1H) ), 4.42 (t, J = 5.6 Hz, 2H), 3.77 (t, J = 5.6 Hz, 2H), 3.44 (q, J = 5.2 Hz, 2H), 1.11 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 183.78, 145.66 (q, J = 1.9 Hz), 137.58, 134.20, 124.73, 120.62 (q, J = 255.0 Hz), 117.81, 113.86 (q, J = 0.8 Hz) ), 113.23, 111.29, 68.25, 66.96, 44.46, 14.94; MS (MALDI-TOF): m/z 335 [M] + ; mp 79℃
제조예 37: 2-브로모-1H-인돌-3-카복스알데히드Preparation 37: 2-bromo-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000079
Figure PCTKR2021007223-appb-img-000079
0℃에서 CH2Cl2(6 mL)에 DMF(1.8 mL)을 넣은 다음, POBr3(5.33 g, 18.6 mmol)의 CH2Cl2(10 mL) 용액을 천천히 넣고 15분 환류하였다. 이후, 옥시인돌(1.03 g, 7.74 mmol)을 조금씩 넣어 주고 1시간 더 환류하였다. 반응용액을 차가운 물에 넣고 20분 교반해서 물층을 분리하였다. 물층을 고체 K2CO3로 중화시킨 다음, 생성된 고체를 여과하였다. 얻어진 고체를 컬럼 크로마토그래피(n-Hex:EA=2:1)로 정제하여 표제 화합물 1.2 g(70%, 연갈색 고체)을 얻었다. DMF (1.8 mL) was added to CH 2 Cl 2 (6 mL) at 0° C., and then a CH 2 Cl 2 (10 mL) solution of POBr 3 (5.33 g, 18.6 mmol) was slowly added and refluxed for 15 minutes. Thereafter, oxyindole (1.03 g, 7.74 mmol) was added little by little, and the mixture was refluxed for 1 hour. The reaction solution was put in cold water and stirred for 20 minutes to separate the water layer. The aqueous layer was neutralized with solid K 2 CO 3 , and then the resulting solid was filtered. The obtained solid was purified by column chromatography (n-Hex:EA=2:1) to obtain 1.2 g (70%, light brown solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 13.04 (br s, 1H), 9.90 (s, 1H), 8.08 (m, 1H), 7.43 (m, 1H), 7.29-7.21 (m, 2H) 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.04 (br s, 1H), 9.90 (s, 1H), 8.08 (m, 1H), 7.43 (m, 1H), 7.29-7.21 (m, 2H) )
제조예 38: 에틸 2-(2-브로모-3-포밀-1H-인돌-1-일)아세테이트Preparation 38: ethyl 2- (2-bromo-3-formyl-1H-indol-1-yl) acetate
Figure PCTKR2021007223-appb-img-000080
Figure PCTKR2021007223-appb-img-000080
0℃에서 상기 제조예 37에서 얻은 2-브로모-1H-인돌-3-카복스알데히드(120.0 mg, 0.54 mmol)와 NaH(32.4 mg, 0.81 mmol, 오일 중 60%)에 THF(5 mL)와 DMF(2 mL)를 넣고 5분 교반한 후, 에틸 브로모아세테이트(72.1 μL, 0.65 mmol)를 넣고 실온에서 15시간 교반하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=20:1)로 분리하여 표제 화합물 91 mg(수율 54%, 흰색 고체)을 얻었다. THF (5 mL) in 2-bromo-1H-indole-3-carboxaldehyde (120.0 mg, 0.54 mmol) and NaH (32.4 mg, 0.81 mmol, 60% in oil) obtained in Preparation 37 at 0 ° C. and DMF (2 mL) were added and stirred for 5 minutes, ethyl bromoacetate (72.1 μL, 0.65 mmol) was added, and the mixture was stirred at room temperature for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH 2 Cl 2 :EA=20:1) to obtain 91 mg (yield 54%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.07 (s, 1H), 8.33 (m, 1H), 7.34-7.31 (m, 2H), 7.25 (m, 1H), 4.99 (s, 2H), 4.26 (q, J = 7.2 Hz, 2H), 1.28 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 185.45, 166.55, 137.11, 126.09, 125.18, 124.49, 123.62, 121.40, 116.07, 109.21, 62.35, 46.27, 14.06; MS (MALDI-TOF): m/z 309 [M]+; mp 94℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.07 (s, 1H), 8.33 (m, 1H), 7.34-7.31 (m, 2H), 7.25 (m, 1H), 4.99 (s, 2H), 4.26 (q, J = 7.2 Hz, 2H), 1.28 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 185.45, 166.55, 137.11, 126.09, 125.18, 124.49, 123.62, 121.40, 116.07, 109.21, 62.35, 46.27, 14.06; MS (MALDI-TOF): m/z 309 [M] + ; mp 94℃
제조예 39: 2-브로모-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드Preparation 39: 2-bromo-1- (2-ethoxyethyl) -1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000081
Figure PCTKR2021007223-appb-img-000081
상기 제조예 37에서 얻은 2-브로모-1H-인돌-3-카복스알데히드(400.0 mg, 1.79 mmol)를 DMF(10 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(240.1 μL, 2.15 mmol)와 Cs2CO3(1.75 g, 5.37 mmol)를 넣고 70℃에서 6시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=20:1)로 분리하여 표제 화합물 436 mg(수율 82%, 흰색 고체)을 얻었다. After dissolving 2-bromo-1H-indole-3-carboxaldehyde (400.0 mg, 1.79 mmol) obtained in Preparation Example 37 in DMF (10 mL), 2-bromoethyl ethyl ether (240.1 μL, 2.15 mmol) ) and Cs 2 CO 3 (1.75 g, 5.37 mmol) were added and heated at 70° C. for 6 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (CH 2 Cl 2 :EA=20:1) to obtain 436 mg (yield 82%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.06 (s, 1H), 8.32 (m, 1H), 7.43 (m, 1H), 7.34-7.30 (m, 2H), 4.46 (t, J = 5.6 Hz, 2H), 3.77 (t, J = 5.6 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 185.61, 137.43, 126.59, 125.43, 124.25, 123.57, 121.30, 115.60, 110.47, 68.40, 67.14, 45.59, 15.17; MS (MALDI-TOF): m/z 295 [M]+; mp 56-57℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.06 (s, 1H), 8.32 (m, 1H), 7.43 (m, 1H), 7.34-7.30 (m, 2H), 4.46 (t, J = 5.6 Hz) , 2H), 3.77 (t, J = 5.6 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 185.61, 137.43, 126.59, 125.43, 124.25, 123.57, 121.30, 115.60, 110.47, 68.40, 67.14, 45.59, 15.17; MS (MALDI-TOF): m/z 295 [M] + ; mp 56-57℃
제조예 40: 2-(트리플루오로메틸)-1H-인돌-3-카복스알데히드Preparation 40: 2- (trifluoromethyl) -1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000082
Figure PCTKR2021007223-appb-img-000082
0℃에서 DMF(10 mL)에 POCl3(1.83 mL g, 20.0 mmol)를 넣고 10분 교반한 후, 2-트리플루오로메틸인돌(740.6 mg, 4.0 mml)의 DMF(10 mL) 용액을 넣고 80℃에서 5시간 가열하였다. 반응용액에 1 N NaOH를 넣어 알칼리화한 후, EA로 추출하였다. 유기층을 브라인(brine) 용액으로 세척하고 Na2SO4로 건조, 여과한 후, 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=7:1)로 분리하여 표제 화합물 324 mg(수율 38%, 흰색 고체)을 얻었다. POCl 3 (1.83 mL g, 20.0 mmol) was added to DMF (10 mL) at 0° C. and stirred for 10 minutes, and a DMF (10 mL) solution of 2-trifluoromethylindole (740.6 mg, 4.0 mml) was added. It was heated at 80 °C for 5 hours. The reaction solution was alkalized by adding 1 N NaOH, followed by extraction with EA. The organic layer was washed with a brine solution, dried over Na 2 SO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=7:1) to obtain 324 mg (yield 38%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 13.42 (br s, 1H), 10.24 (s, 1H), 8.25 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.44 (m, 1H), 7.36 (m, 1H); mp 171-173℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.42 (br s, 1H), 10.24 (s, 1H), 8.25 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.44 (m, 1H), 7.36 (m, 1H); mp 171-173℃
제조예 41: 1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-카복스알데히드Preparation 41: 1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indole-3-carboxaldehyde
Figure PCTKR2021007223-appb-img-000083
Figure PCTKR2021007223-appb-img-000083
2-(트리플루오로메틸)-1H-인돌-3-카복스알데히드(350.0 mg, 1.64 mmol)를 DMF(10 mL)에 녹인 후, 2-브로모에틸 에틸 에테르(220.0 μL, 1.97 mmol)와 Cs2CO3(1.60 g, 4.92 mmol)를 넣고 70℃에서 15시간 가열하였다. 반응용액을 물로 희석하고 EA로 추출하였다. 유기층을 MgSO4로 건조, 여과하고 감압 증류하였다. 잔류물을 컬럼 크로마토그래피(n-Hex:EA=5:1)로 분리하여 표제 화합물 184 mg(수율 39%, 흰색 고체)을 얻었다. 2-(trifluoromethyl)-1H-indole-3-carboxaldehyde (350.0 mg, 1.64 mmol) was dissolved in DMF (10 mL), followed by 2-bromoethyl ethyl ether (220.0 μL, 1.97 mmol) and Cs 2 CO 3 (1.60 g, 4.92 mmol) was added and heated at 70° C. for 15 hours. The reaction solution was diluted with water and extracted with EA. The organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=5:1) to obtain 184 mg (yield 39%, white solid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 10.41 (s, 1H), 8.52 (d, J = 8.0 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.46 (m, 1H), 7.39 (m, 1H), 4.52 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.13 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 185.81, 137.63, 131.27 (q, J = 37.9 Hz), 126.02, 124.30, 124.21, 123.66, 121.29 (q, J = 270.3 Hz), 117.84 (q, J = 1.5 Hz), 111.18, 68.82, 66.93, 45.58 (q, J = 2.5 Hz), 14.94; MS (MALDI-TOF): m/z 286 [M+H]+; mp 46℃ 1 H NMR (400 MHz, CDCl 3 ): δ 10.41 (s, 1H), 8.52 (d, J = 8.0 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.46 (m, 1H), 7.39 (m, 1H), 4.52 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.45 (q, J = 7.2 Hz, 2H), 1.13 (t, J = 7.2) Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 185.81, 137.63, 131.27 (q, J = 37.9 Hz), 126.02, 124.30, 124.21, 123.66, 121.29 (q, J = 270.3 Hz), 117.84 (q, J = 270.3 Hz) 1.5 Hz), 111.18, 68.82, 66.93, 45.58 (q, J = 2.5 Hz), 14.94; MS (MALDI-TOF): m/z 286 [M+H] + ; mp 46℃
<N-아실히드라존 유도체의 제조><Preparation of N-acylhydrazone derivative>
Figure PCTKR2021007223-appb-img-000084
Figure PCTKR2021007223-appb-img-000084
실시예 1: (E)-N'-[(2-클로로-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 1: Preparation of (E)-N'-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000085
Figure PCTKR2021007223-appb-img-000085
5-메틸벤조퓨란-2-카보히드라지드(95.1 mg, 0.50 mmol)와 2-클로로-1H-인돌-3-카복스알데히드(89.8 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 8시간 환류하였다. 반응용액을 감압 증류하고 잔류물을 n-Hex/CH2Cl2(1:1) 혼합용매로 세척하여 표제 화합물 153 mg(수율 87%, 연노란색 고체)을 얻었다.5-Methylbenzofuran-2-carbohydrazide (95.1 mg, 0.50 mmol) and 2-chloro-1H-indole-3-carboxaldehyde (89.8 mg, 0.50 mmol) in 1-PrOH (15 mL) and acetic acid ( 1-2 drops) and refluxed for 8 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with n-Hex/CH 2 Cl 2 (1:1) mixed solvent to obtain 153 mg (yield 87%, pale yellow solid) of the title compound.
1H NMR (300 MHz, DMSO-d6) δ 12.48 (s, 1H), 11.98 (s, 1H), 8.73 (s, 1H), 8.28 (d, J = 7.2 Hz, 1H), 7.63-7.58 (m, 3H), 7.39 (d, J = 7.2 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.27-7.19 (m, 2H), 2.44 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ 154.20, 152.86, 148.41, 142.86, 135.02, 132.91, 128.33, 127.26, 127.16, 124.06, 123.20, 122.24, 121.33, 121.28, 111.37, 111.21, 110.03, 107.39, 20.83; HRMS (TOF MS ES-): m/z calcd for C19H13ClN3O2 (M-H)- 350.0696, found 350.0700; mp 242℃ dec. 1 H NMR (300 MHz, DMSO-d 6 ) δ 12.48 (s, 1H), 11.98 (s, 1H), 8.73 (s, 1H), 8.28 (d, J = 7.2 Hz, 1H), 7.63-7.58 ( m, 3H), 7.39 (d, J = 7.2 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.27-7.19 (m, 2H), 2.44 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ) δ 154.20, 152.86, 148.41, 142.86, 135.02, 132.91, 128.33, 127.26, 127.16, 124.06, 123.20, 122.24, 121.33, 121.28, 111.83, 111.21, 110.83, 111.21, 110.03 ; HRMS (TOF MS ES - ): m/z calcd for C 19 H 13 ClN 3 O 2 (MH) - 350.0696, found 350.0700; mp 242°C dec.
실시예 2: (E)-N'-[(2-클로로-1-메틸-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 2: Preparation of (E)-N'-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000086
Figure PCTKR2021007223-appb-img-000086
5-메틸벤조퓨란-2-카보히드라지드(95.1 mg, 0.50 mmol)와 2-클로로-1-메틸-1H-인돌-3-카복스알데히드(96.8 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, EtOH로 세척하여 표제 화합물 100 mg(수율 55%, 연갈색 고체)을 얻었다.5-Methylbenzofuran-2-carbohydrazide (95.1 mg, 0.50 mmol) and 2-chloro-1-methyl-1H-indole-3-carboxaldehyde (96.8 mg, 0.50 mmol) in 1-PrOH (15 mL) ) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was cooled and the solid was filtered and washed with EtOH to obtain 100 mg (yield 55%, light brown solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.57 (m, 4H), 7.35-7.25 (m, 3H), 3.81 (s, 3H), 2.43 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.22, 152.87, 148.40, 142.93, 136.08, 132.92, 129.29, 128.34, 127.16, 123.24, 123.21, 122.25, 121.67, 121.42, 111.38, 110.34, 110.06, 107.28, 30.19, 20.84; HRMS (TOF MS ES-): m/z calcd for C20H15ClN3O2 (M-H)- 364.0853, found 364.0847; mp 246-247℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.00 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.57 (m, 4H), 7.35 -7.25 (m, 3H), 3.81 (s, 3H), 2.43 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.22, 152.87, 148.40, 142.93, 136.08, 132.92, 129.29, 128.34, 127.16, 123.24, 123.21, 122.25, 121.67, 121.42, 111.38, 110.34, 110.28 30.19, 20.84; HRMS (TOF MS ES - ): m/z calcd for C 20 H 15 ClN 3 O 2 (MH) - 364.0853, found 364.0847; mp 246-247℃
실시예 3: 에틸 (E)-2-{2-클로로-3-[(2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸]-1H-인돌-1-일}아세테이트의 제조Example 3: Ethyl (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl]-1H-indol-1-yl}acetate manufacture of
Figure PCTKR2021007223-appb-img-000087
Figure PCTKR2021007223-appb-img-000087
5-메틸벤조퓨란-2-카보히드라지드(60.9 mg, 0.32 mmol)와 에틸 2-(2-클로로-3-포밀-1H-인돌-1-일)아세테이트(85.0 mg, 0.32 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 30시간 환류하였다. 반응용액을 감압증류하고 잔류물을 Et2O로 세척하여 표제 화합물 85 mg(수율 61%, 흰색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (60.9 mg, 0.32 mmol) in ethyl 2- (2-chloro-3-formyl-1H-indol-1-yl) acetate (85.0 mg, 0.32 mmol) 1- PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 30 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with Et 2 O to obtain 85 mg (yield 61%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.06 (s, 1H), 8.76 (s, 1H), 8.34 (d, J = 6.8 Hz, 1H), 7.64-7.58 (m, 4H), 7.34-7.27 (m, 3H), 5.26 (s, 2H), 4.19 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.22 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 167.84, 154.28, 152.88, 148.33, 142.65, 136.14, 132.94, 129.17, 128.39, 127.15, 123.57, 123.29, 122.27, 121.97, 121.49, 111.39, 110.30, 110.19, 108.22, 61.45, 44.78, 20.84, 14.00; HRMS (TOF MS ES-): m/z calcd for C23H19ClN3O4 (M-H)- 436.1064, found 436.1068; mp 198℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.06 (s, 1H), 8.76 (s, 1H), 8.34 (d, J = 6.8 Hz, 1H), 7.64-7.58 (m, 4H), 7.34 -7.27 (m, 3H), 5.26 (s, 2H), 4.19 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.22 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 167.84, 154.28, 152.88, 148.33, 142.65, 136.14, 132.94, 129.17, 128.39, 127.15, 123.57, 123.29, 122.27, 121.97, 121.49, 111.39, 110.30, 110.39 108.22, 61.45, 44.78, 20.84, 14.00; HRMS (TOF MS ES - ): m/z calcd for C 23 H 19 ClN 3 O 4 (MH) - 436.1064, found 436.1068; mp 198℃
실시예 4: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 4: (E) of -N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide Produce
Figure PCTKR2021007223-appb-img-000088
Figure PCTKR2021007223-appb-img-000088
5-메틸벤조퓨란-2-카보히드라지드(98.9 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(130.9 mg, 0.52 mmol)에 1-PrOH (15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 179 mg(수율 81%, 연노란색 고체)을 얻었다.To 5-methylbenzofuran-2-carbohydrazide (98.9 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (130.9 mg, 0.52 mmol) 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=1:1) to obtain 179 mg (yield 81%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.58 (m, 4H), 7.33-7.24 (m, 3H), 4.46 (t, J = 5.6 Hz, 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 6.8 Hz, 2H), 2.43 (s, 3H), 1.00 (t, J = 6.8 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.23, 152.87, 148.40, 142.99, 135.76, 132.92, 129.26, 128.34, 127,16, 123.35, 123.21, 122.25, 121.67, 121.43, 111.37, 110.67, 110.08, 107.53, 67.85, 65.63, 43.52, 20.83, 14.90; HRMS (TOF MS ES-): m/z calcd for C23H21ClN3O3 (M-H)- 422.1271, found 422.1261; mp 172℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.58 (m, 4H), 7.33 -7.24 (m, 3H), 4.46 (t, J = 5.6 Hz, 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 6.8 Hz, 2H), 2.43 (s, 3H) , 1.00 (t, J = 6.8 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.23, 152.87, 148.40, 142.99, 135.76, 132.92, 129.26, 128.34, 127,16, 123.35, 123.21, 122.25, 121.67, 121.43, 111.08, 110.67, 110.08, 110.67, 107.53, 67.85, 65.63, 43.52, 20.83, 14.90; HRMS (TOF MS ES - ): m/z calcd for C 23 H 21 ClN 3 O 3 (MH) - 422.1271, found 422.1261; mp 172℃
실시예 5: (E)-N'-{[2-브로모-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 5: (E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide manufacture of
Figure PCTKR2021007223-appb-img-000089
Figure PCTKR2021007223-appb-img-000089
5-메틸벤조퓨란-2-카보히드라지드(108.4 mg, 0.57 mmol)와 2-브로모-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(168.8 mg, 0.57 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압 증류하고 소량의 CH2Cl2에 녹인 후, n-Hex 용액에 적하하였다. 생성된 고체를 여과하여 표제 화합물 209 mg(수율 78%, 연노란색 고체)을 얻었다. 5-methylbenzofuran-2-carbohydrazide (108.4 mg, 0.57 mmol) and 2-bromo-1- (2-ethoxyethyl) -1H-indole-3-carboxaldehyde (168.8 mg, 0.57 mmol) 1-PrOH (15 mL) and acetic acid (1-2 drops) were added thereto and refluxed for 24 hours. The reaction solution was distilled under reduced pressure , dissolved in a small amount of CH 2 Cl 2 , and then added dropwise to the n-Hex solution. The resulting solid was filtered to obtain 209 mg of the title compound (yield 78%, pale yellow solid).
1H NMR (400 MHz, DMSO-d6): δ 12.03 (s, 1H), 8.72 (s, 1H), 8.35 (d, J = 7.2 Hz, 1H), 7.63-7.58 (m, 4H), 7.33-7.22 (m, 3H), 4.48 (t, J = 5.6 Hz, 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 H, 2H), 2.44 (s, 3H), 1.01 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.21, 152.86, 148.41, 144.36, 137.00, 132.90, 128.32, 127.16, 124.14, 123.13, 122.23, 121.46, 121.29, 119.80, 111.36, 110.77, 110.43, 110.03, 67.96, 65.69, 44.81, 20.83, 14.91; HRMS (TOF MS ES-): m/z calcd for C23H21BrN3O3 (M-H)- 466.0766, found 466.0776; mp 190℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.03 (s, 1H), 8.72 (s, 1H), 8.35 (d, J = 7.2 Hz, 1H), 7.63-7.58 (m, 4H), 7.33 -7.22 (m, 3H), 4.48 (t, J = 5.6 Hz, 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 H, 2H), 2.44 (s, 3H) , 1.01 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.21, 152.86, 148.41, 144.36, 137.00, 132.90, 128.32, 127.16, 124.14, 123.13, 122.23, 121.46, 121.29, 119.80, 111.36, 110.77, 110.43, 110.77 67.96, 65.69, 44.81, 20.83, 14.91; HRMS (TOF MS ES - ): m/z calcd for C 23 H 21 BrN 3 O 3 (MH) - 466.0766, found 466.0776; mp 190℃
실시예 6: (E)-N'-{[1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 6: (E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000090
Figure PCTKR2021007223-appb-img-000090
5-메틸벤조퓨란-2-카보히드라지드(93.2 mg, 0.49 mmol)와 1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-카복스알데히드(139.8 mg, 0.49 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=2:1)로 분리하여 표제 화합물 176 mg (수율 79%, 흰색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (93.2 mg, 0.49 mmol) and 1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indole-3-carboxaldehyde (139.8 mg , 0.49 mmol), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=2:1) to obtain 176 mg (yield 79%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.24 (s, 1H), 9.00 (s, 1H), 8.63 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.67 (s, 1H), 7.61-7.59 (m, 2H), 7.47 (m, 1H), 7.37-7.32 (m, 2H), 4.54 (t, J = 5.4 Hz, 2H), 3.72 (t, J = 5.4 Hz, 2H), 3.37 (q, J = 7.0 Hz, 2H), 2.44 (s, 3H), 10.00 (t, J = 7.0 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.52, 152.93, 148.17, 142.47, 137.83, 133.01, 128.53, 127.12, 125.66, 125.10 (q, J = 35.8 Hz), 124.20, 123.14, 122.48, 122.32, 121.75 (q, J = 269.0 Hz), 113.45, 111.85, 111.42, 110.56, 68.36, 65.79, 45.01, 20.84, 14.85; 19F NMR (376 MHz, DMSO-d6): δ -52.9 (s, 3F); HRMS (TOF MS ES-): m/z calcd for C24H21F3N3O3 (M-H)- 456.1535, found 456.1531; mp 183℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.24 (s, 1H), 9.00 (s, 1H), 8.63 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H) ), 7.67 (s, 1H), 7.61-7.59 (m, 2H), 7.47 (m, 1H), 7.37-7.32 (m, 2H), 4.54 (t, J = 5.4 Hz, 2H), 3.72 (t, J = 5.4 Hz, 2H), 3.37 (q, J = 7.0 Hz, 2H), 2.44 (s, 3H), 10.00 (t, J = 7.0 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.52, 152.93, 148.17, 142.47, 137.83, 133.01, 128.53, 127.12, 125.66, 125.10 (q, J = 35.8 Hz), 124.20, 123.14, 122.48, 122.32, 121.75 (q, J = 269.0 Hz), 113.45, 111.85, 111.42, 110.56, 68.36, 65.79, 45.01, 20.84, 14.85; 19 F NMR (376 MHz, DMSO-d 6 ): δ -52.9 (s, 3F); HRMS (TOF MS ES -): m / z calcd for C 24 H 21 F 3 N 3 O 3 (MH) - 456.1535, found 456.1531; mp 183℃
실시예 7: (E)-N'-{[2-클로로-1-(2-메톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 7: (E) of -N'-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide Produce
Figure PCTKR2021007223-appb-img-000091
Figure PCTKR2021007223-appb-img-000091
5-메틸벤조퓨란-2-카보히드라지드(319.5 mg, 1.68 mmol)와 2-클로로-1-(2-메톡시에틸)-1H-인돌-3-카복스알데히드(399.3 mg, 1.68 mmol)에 1-PrOH(20 mL)과 아세트산(1-2 방울)을 넣고 15시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, n-Hex로 세척하여 표제 화합물 630 mg(수율 91%, 연노란색 고체)을 얻었다.To 5-methylbenzofuran-2-carbohydrazide (319.5 mg, 1.68 mmol) and 2-chloro-1-(2-methoxyethyl)-1H-indole-3-carboxaldehyde (399.3 mg, 1.68 mmol) 1-PrOH (20 mL) and acetic acid (1-2 drops) were added and refluxed for 15 hours. The reaction solution was cooled, the solid was filtered, and then washed with n-Hex to obtain 630 mg (yield 91%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.33 (d, J = 7.5 Hz, 1H), 7.63-7.58 (m, 4H), 7.33-7.24 (m, 3H), 4.48 (t, J = 5.4 Hz, 2H), 3.68 (t, J = 5.4 Hz, 2H), 3.21 (s, 3H), 2.43 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.23, 152.88, 148.40, 142.97, 135.76, 132.93, 129.20, 128.36, 127.16, 123.34, 123.25, 122.25, 121.68, 121.44, 111.38, 110.68, 110.10, 107.55, 70.11, 58.29, 43.34, 20.84; HRMS (TOF MS ES-): m/z calcd for C22H19ClN3O3 (M-H)- 408.1115, found 408.1099; mp 243℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.33 (d, J = 7.5 Hz, 1H), 7.63-7.58 (m, 4H), 7.33 -7.24 (m, 3H), 4.48 (t, J = 5.4 Hz, 2H), 3.68 (t, J = 5.4 Hz, 2H), 3.21 (s, 3H), 2.43 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.23, 152.88, 148.40, 142.97, 135.76, 132.93, 129.20, 128.36, 127.16, 123.34, 123.25, 122.25, 121.68, 121.44, 111.38, 110.68, 110.10, 110.68 70.11, 58.29, 43.34, 20.84; HRMS (TOF MS ES - ): m/z calcd for C 22 H 19 ClN 3 O 3 (MH) - 408.1115, found 408.1099; mp 243℃
실시예 8: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 8: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000092
Figure PCTKR2021007223-appb-img-000092
5-메틸벤조퓨란-2-카보히드라지드(127.4 mg, 0.67 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드(188.8 mg, 0.67 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:2)로 분리하여 표제 화합물 213 mg(수율 70%, 연노란색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (127.4 mg, 0.67 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (188.8 mg , 0.67 mmol), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=1:2) to obtain 213 mg (yield 70%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.61-7.57 (m, 3H), 7.52 (d, J = 9.2 Hz, 1H), 7.32 (dd, J = 8.8, 1.6 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H), 4.42 (t, J = 5.2 Hz, 2H), 3.82 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (400 MHz, DMSO-d6): δ 155.20, 154.17. 152.85, 148.48, 142.94, 132.91, 130.74, 128.95, 128.31, 127.17, 124.01, 122.23, 112.24, 111.53, 111.35, 110.04, 107.24, 104.11, 67.91, 65.62, 55.33, 43.65, 20.83, 14.90; HRMS (TOF MS ES-): m/z calcd for C24H23ClN3O4 (M-H)- 452.1377, found 452.1355; mp 208℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.61-7.57 (m, 3H), 7.52 (d, J = 9.2 Hz, 1H), 7.32 (dd, J = 8.8, 1.6 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H), 4.42 (t, J = 5.2 Hz, 2H) , 3.82 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.00 (t, J = 7.2 Hz, 3H) ; 13 C NMR (400 MHz, DMSO-d 6 ): δ 155.20, 154.17. 152.85, 148.48, 142.94, 132.91, 130.74, 128.95, 128.31, 127.17, 124.01, 122.23, 112.24, 111.53, 111.35, 110.04, 107.24, 104.11, 67.91, 65.62, 55.33, 43.90, 20.83; HRMS (TOF MS ES - ): m/z calcd for C 24 H 23 ClN 3 O 4 (MH) - 452.1377, found 452.1355; mp 208℃
실시예 9: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 9: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000093
Figure PCTKR2021007223-appb-img-000093
5-메틸벤조퓨란-2-카보히드라지드(95.1 mg, 0.50 mmol)와 2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-카복스알데히드(140.9 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 16시간 환류하였다. 반응용액을 감압 증류하고 소량의 CH2Cl2에 녹인 후, n-Hex 용액에 적하하였다. 생성된 고체를 여과하여 표제 화합물 205 mg(수율 90%, 연노란색 고체)을 얻었다. 5-methylbenzofuran-2-carbohydrazide (95.1 mg, 0.50 mmol) with 2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indole-3-carboxaldehyde (140.9 mg , 0.50 mmol), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 16 hours. The reaction solution was distilled under reduced pressure , dissolved in a small amount of CH 2 Cl 2 , and then added dropwise to the n-Hex solution. The resulting solid was filtered to obtain 205 mg (yield 90%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 11.98 (s, 1H), 8.70 (s, 1H), 8.16 (d, J = 8.8 Hz, 1H), 7.62-7.58 (m, 3H), 7.32 (dd, J = 8.4, 1.2 Hz, 1H), 7.17 (d, J = 2.0 Hz, 1H), 6.90 (dd, J = 8.8, 2.4 Hz, 1H), 4.42 (t, J = 5.6 Hz, 2H), 3.83 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.40 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.02 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 156.75, 154.17. 152.85, 148.39, 142.91, 136.73, 132.91, 128.33, 127.54, 127.15, 122.24, 122.16, 117.27, 111.37, 111.24, 110.03, 107.62, 94.53, 67.88, 65.62, 55.44, 43.41, 20.83, 14.97; HRMS (TOF MS ES-): m/z calcd for C24H23ClN3O4 (M-H)- 452.1377, found 452.1372; mp 210℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.98 (s, 1H), 8.70 (s, 1H), 8.16 (d, J = 8.8 Hz, 1H), 7.62-7.58 (m, 3H), 7.32 (dd, J = 8.4, 1.2 Hz, 1H), 7.17 (d, J = 2.0 Hz, 1H), 6.90 (dd, J = 8.8, 2.4 Hz, 1H), 4.42 (t, J = 5.6 Hz, 2H) , 3.83 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.40 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 1.02 (t, J = 7.2 Hz, 3H) ; 13 C NMR (100 MHz, DMSO-d 6 ): δ 156.75, 154.17. 152.85, 148.39, 142.91, 136.73, 132.91, 128.33, 127.54, 127.15, 122.24, 122.16, 117.27, 111.37, 111.24, 110.03, 107.62, 94.53, 67.88, 65.62, 55.44, 43.41, 20.83; HRMS (TOF MS ES - ): m/z calcd for C 24 H 23 ClN 3 O 4 (MH) - 452.1377, found 452.1372; mp 210℃
실시예 10: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 10: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000094
Figure PCTKR2021007223-appb-img-000094
5-메틸벤조퓨란-2-카보히드라지드(66.6 mg, 0.35 mmol)와 2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-카복스알데히드(94.4 mg, 0.35 mmol)에 1-PrOH(10 mL)과 아세트산(1-2 방울)을 넣고 8시간 환류하였다. 반응용액을 감압 증류하고 소량의 CH2Cl2에 녹인 후, n-Hex 용액에 적하하였다. 생성된 고체를 여과하여 표제 화합물 137 mg (수율 89%, 흰색 고체)을 얻었다. 5-methylbenzofuran-2-carbohydrazide (66.6 mg, 0.35 mmol) with 2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indole-3-carboxaldehyde (94.4 mg , 0.35 mmol), 1-PrOH (10 mL) and acetic acid (1-2 drops) were added and refluxed for 8 hours. The reaction solution was distilled under reduced pressure , dissolved in a small amount of CH 2 Cl 2 , and then added dropwise to the n-Hex solution. The resulting solid was filtered to obtain 137 mg of the title compound (yield 89%, white solid).
1H NMR (400 MHz, DMSO-d6): δ 12.06 (s, 1H), 8.73 (s, 1H), 8.02 (dd, J = 9.6, 2.8 Hz, 1H), 7.67 (dd, J = 9.2, 4.8 Hz, 1H), 7.63-7.58 (m, 3H), 7.32 (d, J = 9.6 Hz, 1H), 7.18 (m, 1H), 4.47 (t, J = 5.6 Hz, 2H), 3.69 (t, J = 5.6 Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 2.43 (s, 3H), 0.99 (t, J = 6.8 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 158.31 (d, J = 233.8 Hz), 154.27, 152.88, 148.31, 142.49, 132.93, 132.47, 130.35, 128.37, 127.14, 123.65 (d, J = 11.3 Hz), 122.24, 112.30 (d, J = 9.4 Hz), 111.36, 111.21 (d, J = 25.9 Hz), 110.19, 107.65 (d, J = 4.4 Hz), 106.37 (d, J = 25.1 Hz), 67.88, 65.64, 43.85, 20.82, 14.88; 19F NMR (376 MHz, DMSO-d6): δ -121.3 (s, 1F); HRMS (TOF MS ES-): m/z calcd for C23H20ClFN3O3 (M-H)- 440.1177, found 440.1185; mp 187℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.06 (s, 1H), 8.73 (s, 1H), 8.02 (dd, J = 9.6, 2.8 Hz, 1H), 7.67 (dd, J = 9.2, 4.8 Hz, 1H), 7.63-7.58 (m, 3H), 7.32 (d, J = 9.6 Hz, 1H), 7.18 (m, 1H), 4.47 (t, J = 5.6 Hz, 2H), 3.69 (t, J = 5.6 Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 2.43 (s, 3H), 0.99 (t, J = 6.8 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 158.31 (d, J = 233.8 Hz), 154.27, 152.88, 148.31, 142.49, 132.93, 132.47, 130.35, 128.37, 127.14, 123.65 (d, J = 11.3 Hz) ), 122.24, 112.30 (d, J = 9.4 Hz), 111.36, 111.21 (d, J = 25.9 Hz), 110.19, 107.65 (d, J = 4.4 Hz), 106.37 (d, J = 25.1 Hz), 67.88, 65.64, 43.85, 20.82, 14.88; 19 F NMR (376 MHz, DMSO-d 6 ): δ -121.3 (s, 1F); HRMS (TOF MS ES - ): m/z calcd for C 23 H 20 ClFN 3 O 3 (MH) - 440.1177, found 440.1185; mp 187℃
실시예 11: (E)-N'-{[2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 11: (E)-N'-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydra manufacture of zeed
Figure PCTKR2021007223-appb-img-000095
Figure PCTKR2021007223-appb-img-000095
5-메틸벤조퓨란-2-카보히드라지드(165.5 mg, 0.87 mmol)와 2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(249.0 mg, 0.87 mmol)에 1-PrOH (30 mL)과 아세트산(1-2 방울)을 넣고 3시간 환류하였다. 반응용액을 감압 증류하고 EtOH로 재결정하였다. 생성된 고체를 여과하고 차가운 EtOH로 세척하여 표제 화합물 270 mg (수율 68%, 흰색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (165.5 mg, 0.87 mmol) and 2,5-dichloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (249.0 mg, 0.87 mmol) ), 1-PrOH (30 mL) and acetic acid (1-2 drops) were added and refluxed for 3 hours. The reaction solution was distilled under reduced pressure and recrystallized from EtOH. The resulting solid was filtered and washed with cold EtOH to obtain 270 mg (yield 68%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.07 (s, 1H), 8.73 (s, 1H), 8.32 (s, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.63-7.58 (m, 3H), 7.35-7.31 (m, 2H), 4.46 (t, J = 5.0 Hz, 2H), 3.69 (t, J = 5.0 Hz, 2H), 3.38 (q, J = 7.0 Hz, 2H), 2.43 (s, 3H), 0.98 (t, J = 7.0 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.31, 152.92, 148.30, 142.52, 134.40, 132.98, 130.52, 128.44, 127.17, 126.42, 124.25, 123.16, 122.30, 120.44, 112.65, 111.42, 110.30, 107.33, 67.89, 65.68, 43.88, 20.88, 14.93; HRMS (ESI): m/z calcd for C23H22Cl2N3O3 (M+H)+ 458.1038, found 458.1037; mp 98℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.07 (s, 1H), 8.73 (s, 1H), 8.32 (s, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.63-7.58 (m, 3H), 7.35-7.31 (m, 2H), 4.46 (t, J = 5.0 Hz, 2H), 3.69 (t, J = 5.0 Hz, 2H), 3.38 (q, J = 7.0 Hz, 2H) , 2.43 (s, 3H), 0.98 (t, J = 7.0 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.31, 152.92, 148.30, 142.52, 134.40, 132.98, 130.52, 128.44, 127.17, 126.42, 124.25, 123.16, 122.30, 120.44, 112.65, 111.42, 110.33 67.89, 65.68, 43.88, 20.88, 14.93; HRMS (ESI): m/z calcd for C 23 H 22 Cl 2 N 3 O 3 (M+H) + 458.1038, found 458.1037; mp 98℃
실시예 12: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 12: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methyl Preparation of benzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000096
Figure PCTKR2021007223-appb-img-000096
5-메틸벤조퓨란-2-카보히드라지드(22.8 mg, 0.12 mmol)와 2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-카복스알데히드(40.3 mg, 0.12 mmol)에 1-PrOH(10 mL)과 아세트산(1-2 방울)을 넣고 18시간 환류하였다. 반응용액을 감압증류하고 잔류물을 컬럼 크로마토그래피(n-Hex:EA=1:2)로 분리하여 표제 화합물 52 mg(수율 85%, 연노란색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (22.8 mg, 0.12 mmol) with 2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indole-3-carbox 1-PrOH (10 mL) and acetic acid (1-2 drops) were added to the aldehyde (40.3 mg, 0.12 mmol) and refluxed for 18 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex:EA=1:2) to obtain 52 mg (yield 85%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.10 (s, 1H), 8.75 (s, 1H), 8.26 (s, 1H), 7.45 (d, J = 9.2 Hz, 1H), 7.63-7.58 (m, 3H), 7.33-7.29 (m, 2H), 4.49 (t, J = 5.2 Hz, 2H), 3.71 (t, J = 5.2 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 2.43 (s, 3H), 0.99 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.27, 152.88, 148.29, 143.57, 142.30, 134.32, 132.95, 130.87, 128.41, 127.13, 123.45, 122.27, 120.34 (q, J = 253.9 Hz), 116.68, 113.47, 112.43, 111.37, 110.27, 107.92, 67.84, 65.63, 43.93, 20.83, 14.87; 19F NMR (376 MHz, DMSO-d6): δ -56.8 (s, 3F); HRMS (TOF MS ES-): m/z calcd for C24H20ClF3N3O4 (M-H)- 506.1094, found 506.1087; mp 84-85℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.10 (s, 1H), 8.75 (s, 1H), 8.26 (s, 1H), 7.45 (d, J = 9.2 Hz, 1H), 7.63-7.58 (m, 3H), 7.33-7.29 (m, 2H), 4.49 (t, J = 5.2 Hz, 2H), 3.71 (t, J = 5.2 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H) , 2.43 (s, 3H), 0.99 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.27, 152.88, 148.29, 143.57, 142.30, 134.32, 132.95, 130.87, 128.41, 127.13, 123.45, 122.27, 120.34 (q, J = 253.9 Hz), 116.68, 113.47, 112.43, 111.37, 110.27, 107.92, 67.84, 65.63, 43.93, 20.83, 14.87; 19 F NMR (376 MHz, DMSO-d 6 ): δ -56.8 (s, 3F); HRMS (TOF MS ES - ): m/z calcd for C 24 H 20 ClF 3 N 3 O 4 (MH) - 506.1094, found 506.1087; mp 84-85℃
실시예 13: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드의 제조Example 13: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2- Preparation of Carbohydrazide
Figure PCTKR2021007223-appb-img-000097
Figure PCTKR2021007223-appb-img-000097
5-메틸벤조퓨란-2-카보히드라지드(374.7 mg, 1.97 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-카복스알데히드(523.5 mg, 1.97 mmol)에 1-PrOH(30 mL)과 아세트산(1-2 방울)을 넣고 3시간 가열 환류하였다. 반응용액을 감압 증류하고 EtOH로 재결정하였다. 생성된 고체를 여과하고 차가운 EtOH로 세척하여 표제 화합물 690 mg(수율 80%, 연노란색 고체)을 얻었다.5-methylbenzofuran-2-carbohydrazide (374.7 mg, 1.97 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indole-3-carboxaldehyde (523.5 mg, 1.97 mmol), 1-PrOH (30 mL) and acetic acid (1-2 drops) were added, and the mixture was heated and refluxed for 3 hours. The reaction solution was distilled under reduced pressure and recrystallized from EtOH. The resulting solid was filtered and washed with cold EtOH to give 690 mg (yield 80%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 11.99 (s, 1H), 8.75 (s, 1H), 8.11 (s, 1H), 7.63-7.59 (m, 3H), 7.50 (d, J = 8.4 Hz, 1H), 7.33 (dd, J = 8.4, 1.4 Hz, 1H), 7.14 (dd, J = 8.4, 1.4 Hz, 1H), 4.43 (t, J = 5.4 Hz, 2H), 3.69 (t, J = 5.4 Hz, 2H), 3.39 (q, J = 7.0 Hz, 2H), 2.45 (s, 3H), 2.44 (s, 3H), 1.01 (t, J = 7.0 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.21, 152.90, 148.46, 143.38, 134.16, 132.95, 130.58, 129.22, 128.37, 127.20, 124.64, 123.54, 122.28, 121.08, 111.41, 110.42, 110.10, 107.09, 67.90, 65.66, 43.57, 21.34, 20.88, 14.95; HRMS (TOF MS ES-): m/z calcd for C24H23ClN3O3 (M-H)- 436.1428, found 436.1442; mp 197℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.99 (s, 1H), 8.75 (s, 1H), 8.11 (s, 1H), 7.63-7.59 (m, 3H), 7.50 (d, J = 8.4 Hz, 1H), 7.33 (dd, J = 8.4, 1.4 Hz, 1H), 7.14 (dd, J = 8.4, 1.4 Hz, 1H), 4.43 (t, J = 5.4 Hz, 2H), 3.69 (t, J = 5.4 Hz, 2H), 3.39 (q, J = 7.0 Hz, 2H), 2.45 (s, 3H), 2.44 (s, 3H), 1.01 (t, J = 7.0 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.21, 152.90, 148.46, 143.38, 134.16, 132.95, 130.58, 129.22, 128.37, 127.20, 124.64, 123.54, 122.28, 121.08, 111.41, 110.42, 110.10, 107.09 67.90, 65.66, 43.57, 21.34, 20.88, 14.95; HRMS (TOF MS ES - ): m/z calcd for C 24 H 23 ClN 3 O 3 (MH) - 436.1428, found 436.1442; mp 197℃
실시예 14: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드의 제조Example 14: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydrazide manufacture of
Figure PCTKR2021007223-appb-img-000098
Figure PCTKR2021007223-appb-img-000098
5-메톡시벤조퓨란-2-카보히드라지드(103.1 mg, 0.50 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(125.9 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 7시간 환류하였다. 반응용액을 감압 증류하고 잔류물을 n-Hex/CH2Cl2 혼합용매(1:1)로 세척하여 표제 화합물 204 mg(수율 93%, 연노란색 고체)을 얻었다. 5-Methoxybenzofuran-2-carbohydrazide (103.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (125.9 mg, 0.50 mmol) 1-PrOH (15 mL) and acetic acid (1-2 drops) were added thereto, and the mixture was refluxed for 7 hours. The reaction solution was distilled under reduced pressure, and the residue was washed with n-Hex/CH 2 Cl 2 mixed solvent (1:1) to obtain 204 mg of the title compound (yield 93%, pale yellow solid).
1H NMR (400 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.60 (m, 3H), 7.33-7.24 (m, 3H), 7.10 (dd, J = 8.8, 2.4 Hz, 1H), 4.46 (t, J = 5.6 Hz, 2H), 3.82 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 156.11, 154.16, 149.34, 148.94, 143.01, 135.76, 129.27, 127.72, 123.35, 123.21, 121.68, 121.43, 116.39, 112.47, 110.68, 110.43, 107.53, 104.20, 67.85, 65.63, 55.63, 43.53, 14.90; HRMS (TOF MS ES-): m/z calcd for C23H21ClN3O4 (M-H)- 438.1221, found 438.1218; mp 185℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63-7.60 (m, 3H), 7.33 -7.24 (m, 3H), 7.10 (dd, J = 8.8, 2.4 Hz, 1H), 4.46 (t, J = 5.6 Hz, 2H), 3.82 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 156.11, 154.16, 149.34, 148.94, 143.01, 135.76, 129.27, 127.72, 123.35, 123.21, 121.68, 121.43, 116.39, 112.47, 110.68, 104.20, 107.53, 110.43, 67.85, 65.63, 55.63, 43.53, 14.90; HRMS (TOF MS ES - ): m/z calcd for C 23 H 21 ClN 3 O 4 (MH) - 438.1221, found 438.1218; mp 185℃
실시예 15: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드의 제조Example 15: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran- Preparation of 2-Carbohydrazide
Figure PCTKR2021007223-appb-img-000099
Figure PCTKR2021007223-appb-img-000099
5-메톡시벤조퓨란-2-카보히드라지드(103.1 mg, 0.50 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드(140.9 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 8시간 환류하였다. 반응용액을 감압증류하고 잔류물을 컬럼 크로마토그래피(n-Hex-EA=2:3)로 분리하여 표제 화합물 222 mg(수율 94%, 연노란색 고체)을 얻었다.5-Methoxybenzofuran-2-carbohydrazide (103.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (140.9 mg, 0.50 mmol) was added with 1-PrOH (15 mL) and acetic acid (1-2 drops) and refluxed for 8 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (n-Hex-EA=2:3) to obtain 222 mg of the title compound (yield 94%, pale yellow solid).
1H NMR (400 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.62 -7.59 (m, 2H), 7.52 (d, J = 8.8 Hz, 1H), 7.32 (d, J = 2.4 Hz, 1H), 7.09 (dd, J = 8.8, 2.4 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H), 4.42 (t, J = 5.2 Hz, 2H), 3.82 (s, 3H), 3.817 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 156.11, 155.21, 154.12, 149.33, 149.03, 142.97, 130.75, 128.96, 127.74, 124.02, 116.37, 112.45, 112.27, 111.54, 110.42, 107.25, 104.20, 104.09, 67.92, 65.63, 55.63, 55.33, 43.67, 14.91; HRMS (TOF MS ES-): m/z calcd for C24H23ClN3O5 (M-H)- 468.1326, found 468.1322; mp 128-129℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.73 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.62 -7.59 (m, 2H), 7.52 (d, J = 8.8 Hz, 1H), 7.32 (d, J = 2.4 Hz, 1H), 7.09 (dd, J = 8.8, 2.4 Hz, 1H), 6.95 (dd, J = 8.8, 2.4 Hz, 1H) , 4.42 (t, J = 5.2 Hz, 2H), 3.82 (s, 3H), 3.817 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H) , 1.00 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 156.11, 155.21, 154.12, 149.33, 149.03, 142.97, 130.75, 128.96, 127.74, 124.02, 116.37, 112.45, 112.27, 111.54, 110.42, 107.25, 104.20, 104.09 67.92, 65.63, 55.63, 55.33, 43.67, 14.91; HRMS (TOF MS ES - ): m/z calcd for C 24 H 23 ClN 3 O 5 (MH) - 468.1326, found 468.1322; mp 128-129℃
실시예 16: (E)-5-클로로-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}벤조퓨란-2-카보히드라지드의 제조Example 16: of (E)-5-chloro-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbohydrazide Produce
Figure PCTKR2021007223-appb-img-000100
Figure PCTKR2021007223-appb-img-000100
5-클로로벤조퓨란-2-카보히드라지드(103.2 mg, 0.49 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(123.3 mg, 0.49 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압 증류하고 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA= 5:1)로 분리하여 표제 화합물 188 mg(수율 86%, 연노란색 고체)을 얻었다.To 5-chlorobenzofuran-2-carbohydrazide (103.2 mg, 0.49 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (123.3 mg, 0.49 mmol) 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (CH 2 Cl 2 :EA=5:1) to obtain 188 mg (yield 86%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.10 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.94 (d, J = 2.0 Hz, 1H), 7.76 (d, J = 9.2 Hz, 1H), 7.69 (s, 1H), 7.61 (d, J = 7.6 Hz, 1H), 7.53 (dd, J = 9.2, 2.0 Hz, 1H), 7.33-7.24 (m, 2H), 4.46 (t, J = 5.2 Hz, 2H), 3.70 (t, J = 5.2 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 153.79, 152.86, 149.68, 143.29, 135.76, 129.43, 128.66, 128.15, 126.97, 123.33, 123.23, 122.21, 121.72, 121.39, 113.52, 110.71, 109.75, 107.44, 67.84, 65.63, 43.55, 14.90; HRMS (ESI): m/z calcd for C22H20Cl2N3O3 (M+H)+ 444.0882, found 444.0884; mp 117℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.10 (s, 1H), 8.75 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.94 (d, J = 2.0 Hz, 1H) ), 7.76 (d, J = 9.2 Hz, 1H), 7.69 (s, 1H), 7.61 (d, J = 7.6 Hz, 1H), 7.53 (dd, J = 9.2, 2.0 Hz, 1H), 7.33-7.24 (m, 2H), 4.46 (t, J = 5.2 Hz, 2H), 3.70 (t, J = 5.2 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz) , 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 153.79, 152.86, 149.68, 143.29, 135.76, 129.43, 128.66, 128.15, 126.97, 123.33, 123.23, 122.21, 121.72, 121.39, 113.52, 110.71, 109.44 67.84, 65.63, 43.55, 14.90; HRMS (ESI): m/z calcd for C 22 H 20 Cl 2 N 3 O 3 (M+H) + 444.0882, found 444.0884; mp 117℃
실시예 17: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,7-디메틸벤조퓨란-2-카보히드라지드의 제조Example 17: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbohydra manufacture of zeed
Figure PCTKR2021007223-appb-img-000101
Figure PCTKR2021007223-appb-img-000101
4,7-디메틸벤조퓨란-2-카보히드라지드(81.7 mg, 0.40 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(100.7 mg, 0.40 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 24시간 환류하였다. 반응용액을 감압 증류하고 잔류물을 컬럼 크로마토그래피(CH2Cl2:EA=5:1)로 분리하여 표제 화합물 158 mg(수율 90%, 흰색 고체)을 얻었다.4,7-dimethylbenzofuran-2-carbohydrazide (81.7 mg, 0.40 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (100.7 mg, 0.40 mmol) ), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the residue was separated by column chromatography (CH 2 Cl 2 :EA=5:1) to obtain 158 mg (yield 90%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 11.87 (s, 1H), 8.76 (s, 1H), 8.34 (d, J = 7.2 Hz, 1H), 7.76 (s, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.19 (d, J = 8.0 Hz, 1H), 7.05 (d, J = 7.2 Hz, 1H), 4.46 (t, J = 5.6 Hz, 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 2.54 (s, 3H), 2.51 (s, 3H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 154.34, 153.35, 147.46, 142.86, 135.76, 129.60, 129.25, 127.53, 126.50, 123.81, 123.35, 123.22, 121.67, 121.42, 118.71, 110.69, 109.48, 107.50, 67.85, 65.62, 43.53, 17.88, 14.90, 14.52; HRMS (ESI): m/z calcd for C24H25ClN3O3 (M+H)+ 438.1584, found 438.1584; mp 202℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.87 (s, 1H), 8.76 (s, 1H), 8.34 (d, J = 7.2 Hz, 1H), 7.76 (s, 1H), 7.61 (d) , J = 8.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.19 (d, J = 8.0 Hz, 1H), 7.05 (d, J = 7.2 Hz, 1H), 4.46 (t, J = 5.6 Hz) , 2H), 3.71 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 2.54 (s, 3H), 2.51 (s, 3H), 1.00 (t, J = 7.2 Hz) , 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.34, 153.35, 147.46, 142.86, 135.76, 129.60, 129.25, 127.53, 126.50, 123.81, 123.35, 123.22, 121.67, 121.42, 118.71, 110.69, 118.71, 110.69 67.85, 65.62, 43.53, 17.88, 14.90, 14.52; HRMS (ESI): m/z calcd for C 24 H 25 ClN 3 O 3 (M+H) + 438.1584, found 438.1584; mp 202℃
실시예 18: (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,6-디메톡시벤조퓨란-2-카보히드라지드의 제조Example 18: (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbo Preparation of hydrazide
Figure PCTKR2021007223-appb-img-000102
Figure PCTKR2021007223-appb-img-000102
4,6-디메톡시벤조퓨란-2-카보히드라지드(118.1 mg, 0.50 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(125.9 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 4시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, EtOH로 세척하여 표제 화합물 210 mg(수율 89%, 흰색 고체)을 얻었다.4,6-dimethoxybenzofuran-2-carbohydrazide (118.1 mg, 0.50 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (125.9 mg, 0.50) mmol), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 4 hours. The reaction solution was cooled and the solid was filtered and washed with EtOH to obtain 210 mg (yield 89%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 11.82 (s, 1H), 8.69 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63 (s, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.33-7.23 (m, 2H), 6.87 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 4.45 (t, J = 5.6 Hz, 2H), 3.92 (s, 3H), 3.85 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 161.07, 156.44, 154.11, 146.03, 142.31, 135.74, 129.04, 123.33, 123.18, 121.60, 121.41, 110.98, 110.65, 107.58, 107.53, 95.13, 88.44, 67.85, 65.62, 55.84, 43.50, 14.90; HRMS (ESI): m/z calcd for C24H25ClN3O5 (M+H)+ 470.1483, found 470.1482; mp 215℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.82 (s, 1H), 8.69 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.63 (s, 1H), 7.60 (d , J = 7.6 Hz, 1H), 7.33-7.23 (m, 2H), 6.87 (s, 1H), 6.50 (d, J = 2.0 Hz, 1H), 4.45 (t, J = 5.6 Hz, 2H), 3.92 (s, 3H), 3.85 (s, 3H), 3.70 (t, J = 5.6 Hz, 2H), 3.38 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 161.07, 156.44, 154.11, 146.03, 142.31, 135.74, 129.04, 123.33, 123.18, 121.60, 121.41, 110.98, 110.65, 107.58, 107.53, 95.13, 88.85 65.62, 55.84, 43.50, 14.90; HRMS (ESI): m/z calcd for C 24 H 25 ClN 3 O 5 (M+H) + 470.1483, found 470.1482; mp 215℃
실시예 19: (E)-N'-{2-[2-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌]히드라진-1-카보닐}벤조퓨란-5-일)아세트아미드의 제조Example 19: (E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl} Preparation of benzofuran-5-yl)acetamide
Figure PCTKR2021007223-appb-img-000103
Figure PCTKR2021007223-appb-img-000103
N-(2-(히드라진카보닐)벤조퓨란-5-일)아세트아미드(116.6 mg, 0.50 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(125.9 mg, 0.50 mmol)에 1-PrOH(20 mL)과 아세트산(1-2 방울)을 넣고 12시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, EtOH로 세척하여 표제 화합물 166 mg (71%, 연노란색 고체)을 얻었다.N-(2-(hydrazinecarbonyl)benzofuran-5-yl)acetamide (116.6 mg, 0.50 mmol) with 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde 1-PrOH (20 mL) and acetic acid (1-2 drops) were added to (125.9 mg, 0.50 mmol) and refluxed for 12 hours. The reaction solution was cooled and the solid was filtered and washed with EtOH to obtain 166 mg (71%, pale yellow solid) of the title compound.
1H NMR (400 MHz, DMSO-d6): δ 12.02 (s, 1H), 10.07 (s, 1H), 8.76 (s, 1H), 8.32 (d, J = 7.2 Hz, 1H), 8.17 (d, J = 2.4 Hz, 1H), 7.68 (s, 1H), 7.65-7.60 (m, 2H), 7.53 (dd, J = 9.2, 2.4 Hz, 1H), 7.33-7.24 (m, 2H), 4.46 (t, J = 5.6 Hz, 2H), 3.70 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 2.08 (s, 3H), 1.00 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d6): δ 168.24, 154.15, 150.62, 148.84, 143.08, 135.77, 135.54, 129.32, 127.18, 123.36, 123.24, 121.70, 121.44, 119.61, 112.10, 111.83, 110.72, 110.66, 107.53, 67.88, 65.65, 43.55, 23.97, 14.94; HRMS (TOF MS ES-): m/z calcd for C24H22ClN4O4 (M-H)- 465.1330, found 465.1323; mp 226℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 12.02 (s, 1H), 10.07 (s, 1H), 8.76 (s, 1H), 8.32 (d, J = 7.2 Hz, 1H), 8.17 (d) , J = 2.4 Hz, 1H), 7.68 (s, 1H), 7.65-7.60 (m, 2H), 7.53 (dd, J = 9.2, 2.4 Hz, 1H), 7.33-7.24 (m, 2H), 4.46 ( t, J = 5.6 Hz, 2H), 3.70 (t, J = 5.6 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 2.08 (s, 3H), 1.00 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 168.24, 154.15, 150.62, 148.84, 143.08, 135.77, 135.54, 129.32, 127.18, 123.36, 123.24, 121.70, 121.44, 119.661, 112.10, 111.83, 110.72, 110.83 107.53, 67.88, 65.65, 43.55, 23.97, 14.94; HRMS (TOF MS ES - ): m/z calcd for C 24 H 22 ClN 4 O 4 (MH) - 465.1330, found 465.1323; mp 226℃
실시예 20: (E)-에틸-2-(3-((2-(4,6-디메톡시벤조퓨란-2-카보닐)히드라진일리덴)메틸)-2-메틸-1H-인돌-1-일)아세테이트의 제조Example 20: (E)-ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazinylidene)methyl)-2-methyl-1H-indole-1 -Il) Preparation of acetate
Figure PCTKR2021007223-appb-img-000104
Figure PCTKR2021007223-appb-img-000104
4,6-디메톡시벤조퓨란-2-카보히드라지드(118.1 mg, 0.50 mmol)와 에틸 2-(3-포밀-2-메틸-1H-인돌-1-일)아세테이트(122.6 mg, 0.50 mmol)에 1-PrOH(15 mL)과 아세트산(1-2 방울)을 넣고 4시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, EtOH로 세척하여 표제 화합물 197 mg(수율 85%, 흰색 고체)을 얻었다.4,6-dimethoxybenzofuran-2-carbohydrazide (118.1 mg, 0.50 mmol) and ethyl 2-(3-formyl-2-methyl-1H-indol-1-yl)acetate (122.6 mg, 0.50 mmol) 1-PrOH (15 mL) and acetic acid (1-2 drops) were added thereto, and the mixture was refluxed for 4 hours. The reaction solution was cooled and the solid was filtered and washed with EtOH to obtain 197 mg (yield 85%, white solid) of the title compound.
HRMS (ESI): m/z calcd for C25H25N3O6Na(M+Na) 486.1641, found 486.1642 HRMS (ESI): m/z calcd for C 25 H 25 N 3 O 6 Na(M+Na) 486.1641, found 486.1642
실시예 21: (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드의 제조Example 21: (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000105
Figure PCTKR2021007223-appb-img-000105
벤조[b]티오펜-2-카보히드라지드(100 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드(146.6 mg, 0.52 mmol)에 1-PrOH (10 mL)과 아세트산 (1-2 방울)를 넣고 20시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, CH2Cl2로 세척하여 목적화합물 166 mg (70%)을 얻었다. Benzo[b]thiophene-2-carbohydrazide (100 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (146.6 mg, 0.52 mmol) was added with 1-PrOH (10 mL) and acetic acid (1-2 drops) and refluxed for 20 hours. The reaction solution was cooled and the solid was filtered and washed with CH 2 Cl 2 to obtain 166 mg (70%) of the target compound.
1H NMR (400 MHz, DMSO-d 6) δ 12.02 (s, 1H), 8.66 (s, 1H), 8.42 (d, J = 3.6 Hz, 1H), 8.23 (s, 1H), 8.07 (d, J = 7.2 Hz, 1H), 8.01 (d, J = 7.2 Hz, 1H), 7.98-7.89 (m, 1H), 7.56-7.47 (m, 2H), 6.95-6.93 (m, 1H), 4.43 (t, J = 5.2 Hz, 2H), 3.81 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 457 [M+H]+ 1 H NMR (400 MHz, DMSO- d 6 ) δ 12.02 (s, 1H), 8.66 (s, 1H), 8.42 (d, J = 3.6 Hz, 1H), 8.23 (s, 1H), 8.07 (d, J = 7.2 Hz, 1H), 8.01 (d, J = 7.2 Hz, 1H), 7.98-7.89 (m, 1H), 7.56-7.47 (m, 2H), 6.95-6.93 (m, 1H), 4.43 (t) , J = 5.2 Hz, 2H), 3.81 (s, 3H), 3.68 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H) ); MS (MALDI-TOF): m/z 457 [M+H] +
실시예 22: (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드의 제조Example 22: (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2- Preparation of Carbohydrazide
Figure PCTKR2021007223-appb-img-000106
Figure PCTKR2021007223-appb-img-000106
5-페닐퓨란-2-카보히드라지드(105 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드(146.6 mg, 0.52 mmol)에 1-PrOH (10 mL)과 아세트산 (1-2 방울)를 넣고 20시간 환류하였다. 반응용액을 감압농축시키고 잔류물을 컬럼 크로마토그래피(n-Hex-EA=3:1)로 분리하여 표제 화합물 213 mg(수율 88%)을 얻었다.5-phenylfuran-2-carbohydrazide (105 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (146.6 mg, 0.52 mmol), 1-PrOH (10 mL) and acetic acid (1-2 drops) were added and refluxed for 20 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by column chromatography (n-Hex-EA=3:1) to obtain 213 mg (yield 88%) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 11.77 (s, 1H), 8.62 (s, 1H), 7.91-7.83 (m, 2H), 7.74 (m, 2H), 7.62-7.34 (m, 5H), 6.94 (t, J = 8.4 Hz, 1H), 4.42 (t, J = 5.2 Hz, 2H), 3.80 (s, 3H), 3.66 (t, J = 5.2 Hz, 2H), 3.36 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 467 [M+H]+ 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.77 (s, 1H), 8.62 (s, 1H), 7.91-7.83 (m, 2H), 7.74 (m, 2H), 7.62-7.34 (m, 5H) ), 6.94 (t, J = 8.4 Hz, 1H), 4.42 (t, J = 5.2 Hz, 2H), 3.80 (s, 3H), 3.66 (t, J = 5.2 Hz, 2H), 3.36 (q, J) = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 467 [M+H] +
실시예 23: (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드의 제조Example 23: (E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2 -Preparation of carbohydrazide
Figure PCTKR2021007223-appb-img-000107
Figure PCTKR2021007223-appb-img-000107
5-페닐티오펜-2-카보히드라지드(114 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-카복스알데히드(146.6 mg, 0.52 mmol)에 1-PrOH (10 mL)과 아세트산 (1-2 방울)를 넣고 20시간 환류하였다. 반응용액을 감압 농축시키고 잔류물을 컬럼 크로마토그래피(n-Hex-EA=2:1)로 분리하여 표제 화합물 213 mg(수율 85%)을 얻었다.5-phenylthiophene-2-carbohydrazide (114 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indole-3-carboxaldehyde (146.6 mg , 0.52 mmol), 1-PrOH (10 mL) and acetic acid (1-2 drops) were added and refluxed for 20 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by column chromatography (n-Hex-EA=2:1) to obtain 213 mg (yield 85%) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 11.78 (s, 1H), 8.63 (s, 1H), 7.93-7.89 (m, 2H), 7.75 (m, 2H), 7.63-7.39 (m, 5H), 6.95 (t, J = 8.4 Hz, 1H), 4.43 (t, J = 5.2 Hz, 2H), 3.81 (s, 3H), 3.67 (t, J = 5.2 Hz, 2H), 3.37 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 483 [M+H]+ 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.78 (s, 1H), 8.63 (s, 1H), 7.93-7.89 (m, 2H), 7.75 (m, 2H), 7.63-7.39 (m, 5H) ), 6.95 (t, J = 8.4 Hz, 1H), 4.43 (t, J = 5.2 Hz, 2H), 3.81 (s, 3H), 3.67 (t, J = 5.2 Hz, 2H), 3.37 (q, J) = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 483 [M+H] +
실시예 24: (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드의 제조Example 24: (E) of -N'-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide Produce
Figure PCTKR2021007223-appb-img-000108
Figure PCTKR2021007223-appb-img-000108
5-페닐티오펜-2-카보히드라지드(113.4 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(130.8 mg, 0.52 mmol)에 1-PrOH (10 mL)과 아세트산(1-2 방울)를 넣고 20시간 환류하였다. 반응용액을 감압 농축시키고 잔유물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 192 mg (82%)을 얻었다.To 5-phenylthiophene-2-carbohydrazide (113.4 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (130.8 mg, 0.52 mmol) 1-PrOH (10 mL) and acetic acid (1-2 drops) were added and refluxed for 20 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by column chromatography (n -Hex:EA=1:1) to obtain 192 mg (82%) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 11.79 (s, 1H), 8.65 (s, 1H), 8.31 (d, J = 7.6 Hz, 1H), 7.91 (d, J = 3.2 Hz, 1H), 7.76 (m, 2H), 7.62 (m, 2H), 7.48 (m, 2H), 7.41-7.23 (m, 3H), 4.46 (t, J = 5.2 Hz, 2H), 3.70 (t, J = 5.2 Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 453 [M+H]+ 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.79 (s, 1H), 8.65 (s, 1H), 8.31 (d, J = 7.6 Hz, 1H), 7.91 (d, J = 3.2 Hz, 1H) , 7.76 (m, 2H), 7.62 (m, 2H), 7.48 (m, 2H), 7.41-7.23 (m, 3H), 4.46 (t, J = 5.2 Hz, 2H), 3.70 (t, J = 5.2) Hz, 2H), 3.38 (q, J = 6.8 Hz, 2H), 1.00 (t, J = 6.8 Hz, 3H); MS (MALDI-TOF): m/z 453 [M+H] +
실시예 25: (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드의 제조Example 25: (E)-N'-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbo Preparation of hydrazide
Figure PCTKR2021007223-appb-img-000109
Figure PCTKR2021007223-appb-img-000109
1-페닐-1H-피라졸-5-카보히드라지드(105 mg, 0.52 mmol)와 2-클로로-1-(2-에톡시에틸)-1H-인돌-3-카복스알데히드(130.8 mg, 0.52 mmol)에 1-PrOH (10 mL)과 아세트산(1-2 방울)를 넣고 20시간 환류하였다. 반응용액을 감압 농축시키고 잔유물을 컬럼 크로마토그래피(n-Hex:EA=1:1)로 분리하여 표제 화합물 165 mg (73%)을 얻었다.1-phenyl-1H-pyrazole-5-carbohydrazide (105 mg, 0.52 mmol) and 2-chloro-1-(2-ethoxyethyl)-1H-indole-3-carboxaldehyde (130.8 mg, 0.52) mmol), 1-PrOH (10 mL) and acetic acid (1-2 drops) were added and refluxed for 20 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by column chromatography (n -Hex:EA=1:1) to obtain 165 mg (73%) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 11.79 (s, 1H), 8.65 (s, 1H), 8.37 (d, J = 7.6 Hz, 1H), 8.30 (d, J = 7.6 Hz, 1H), 7.91 (d, J = 3.2 Hz, 1H), 7.77-7.60 (m, 4H), 7.49-7.23 (m, 4H), 4.48 (t, J = 5.2 Hz, 2H), 3.71 (t, J = 5.2 Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); MS (MALDI-TOF): m/z 437 [M+H]+ 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.79 (s, 1H), 8.65 (s, 1H), 8.37 (d, J = 7.6 Hz, 1H), 8.30 (d, J = 7.6 Hz, 1H) , 7.91 (d, J = 3.2 Hz, 1H), 7.77-7.60 (m, 4H), 7.49-7.23 (m, 4H), 4.48 (t, J = 5.2 Hz, 2H), 3.71 (t, J = 5.2) Hz, 2H), 3.39 (q, J = 7.2 Hz, 2H), 1.00 (t, J = 7.2 Hz, 3H); MS (MALDI-TOF): m/z 437 [M+H] +
실시예 26: (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트의 제조Example 26: (E)-ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indole- 1-yl) Preparation of acetate
Figure PCTKR2021007223-appb-img-000110
Figure PCTKR2021007223-appb-img-000110
나프토[2,1-b]퓨란-2-카보히드라지드(113.1 mg, 0.50 mmol)와 에틸 2-(2-클로로-3-포밀-1H-인돌-1-일)아세테이트(132.8 mg, 0.50 mmol)에 1-PrOH (15 mL)과 아세트산(1-2 방울)를 넣고 24시간 환류하였다. 반응용액을 냉각시키고 고체를 여과한 후, EtOH로 세척하여 표제 화합물 186 mg (78%, 흰색 고체)을 얻었다. Naphtho [2,1-b] furan-2-carbohydrazide (113.1 mg, 0.50 mmol) and ethyl 2- (2-chloro-3-formyl- 1H -indol-1-yl) acetate (132.8 mg, 0.50 mmol), 1-PrOH (15 mL) and acetic acid (1-2 drops) were added and refluxed for 24 hours. The reaction solution was cooled and the solid was filtered and washed with EtOH to obtain 186 mg (78%, white solid) of the title compound.
1H NMR (400 MHz, DMSO-d 6) δ 12.16 (s, 1H), 8.80 (s, 1H), 8.42 (d, J = 8.0 Hz, 1H), 8.39-8.36 (m, 2H), 8.11 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 9.2 Hz, 1H), 7.89 (d, J = 9.2 Hz, 1H), 7.72 (m, 1H), 7.63-7.59 (m, 2H), 7.35-7.28 (m, 2H), 5.27 (s, 2H), 4.20 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, DMSO-d 6) δ 167.85, 154.19, 152.34, 147.78, 142.58, 136.15, 130.11, 129.15, 128.86, 128.36, 127.49, 127.32, 125.43, 123.70, 123.58, 123.29, 122.69, 121.96, 121.51, 112.51, 110.31, 110.02, 108.24, 61.45, 44.79, 14.00; HRMS (TOF MS ES-): m/z calcd for C26H19ClN3O4 (M-H)- 472.1064, found 472.1068; mp 231-232℃ 1 H NMR (400 MHz, DMSO- d 6 ) δ 12.16 (s, 1H), 8.80 (s, 1H), 8.42 (d, J = 8.0 Hz, 1H), 8.39-8.36 (m, 2H), 8.11 ( d, J = 8.0 Hz, 1H), 8.04 (d, J = 9.2 Hz, 1H), 7.89 (d, J = 9.2 Hz, 1H), 7.72 (m, 1H), 7.63-7.59 (m, 2H), 7.35-7.28 (m, 2H), 5.27 (s, 2H), 4.20 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H); 13 C NMR (100 MHz, DMSO- d 6 ) δ 167.85, 154.19, 152.34, 147.78, 142.58, 136.15, 130.11, 129.15, 128.86, 128.36, 127.49, 127.32, 125.43, 123.70, 123.58, 123.29, 122.69, 121.96 , 112.51, 110.31, 110.02, 108.24, 61.45, 44.79, 14.00; HRMS (TOF MS ES -): m / z calcd for C 26 H 19 ClN 3 O 4 (MH) - 472.1064, found 472.1068; mp 231-232℃
실시예 27: (E)-메틸 2-(2-메틸-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트의 제조Example 27: (E)-methyl 2-(2-methyl-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indole- 1-yl) Preparation of acetate
Figure PCTKR2021007223-appb-img-000111
Figure PCTKR2021007223-appb-img-000111
한국 등록특허 제1790193호에 개시된 합성 방법에 의해 표제 화합물(수율 90 %, 황색 고체)을 얻었다.The title compound (yield 90%, yellow solid) was obtained by the synthesis method disclosed in Korean Patent Registration No. 1790193.
ESIMS found: m/z 440.36[M+H]+, 879.16[2M+H]+.ESIMS found: m/z 440.36[M+H]+, 879.16[2M+H]+.
실시예 28: (E)-에틸 2-(2-메틸-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라조노)메틸)-1H-인돌-1-일)아세테이트의 제조Example 28: (E)-ethyl 2-(2-methyl-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazono)methyl)-1H-indole-1 -Il) Preparation of acetate
Figure PCTKR2021007223-appb-img-000112
Figure PCTKR2021007223-appb-img-000112
한국 공개특허공보 제2014-0128238호에 개시된 합성 방법에 의해 표제 화합물(수율 75%, 연황색 고체)을 얻었다.The title compound (yield 75%, light yellow solid) was obtained by the synthesis method disclosed in Korean Patent Application Laid-Open No. 2014-0128238.
1H NMR (400 MHz, DMSO-d6) δppm:11.838 (1H, s, -NHCO-), 8.862 (1H, s, -N=CH-Ar), 8.438-8.418 (1H, d, J = 8, Ar-H), 8.355-8.307 (2H, dd, J = 8.8, 3.6, Ar-H), 8.114-8.094 (1H, d, J = 8.0, Ar-H), 8.041-8.018 (1H, d, J = 8.0, Ar-H), 7.891-7.869 (1H, d, J = 8.8, Ar-H), 7.731-7.694 (1H, t, J = 7.2, Ar-H), 7.622-7.585 (1H, t, J = 8.0, Ar-H), 7.483-7.461 (1H, t, J = 8.8, Ar-H), 7.217-7.198 (2H, m, Ar-H), 5.192 (2H, s, -N-CH2), 4.211-4.158 (2H, q, J = 7.2, -O-CH2), 2.505 (3H, s, -CH3), 1.249-1.214 (3H, t, J = 7.2, -CH3); 13C NMR (100 MHz, DMSO-d6): δppm:168.602, 153.88, 152.217, 148.131, 145.138, 141.430, 137.031, 130.101, 128.833, 128.178, 127.482, 127.271, 125.388, 124.674, 123.720, 122.755, 122.242, 121.368, 120.985, 112.485, 109.656, 109.539, 108.235, 61.201, 44.431, 14.026, 9.961. ESIMS found: m/z 452.6 [M-H]-, 454.6 [M+H]+, 476.5 [M+Na]+; Rf = 0.50 (n-Hex:EA=1:2) 1 H NMR (400 MHz, DMSO-d6) δ ppm:11.838 (1H, s, -NHCO-), 8.862 (1H, s, -N=CH-Ar), 8.438-8.418 (1H, d, J = 8, Ar-H), 8.355-8.307 (2H, dd, J = 8.8, 3.6, Ar-H), 8.114-8.094 (1H, d, J = 8.0, Ar-H), 8.041-8.018 (1H, d, J = 8.0, Ar-H), 7.891-7.869 (1H, d, J = 8.8, Ar-H), 7.731-7.694 (1H, t, J = 7.2, Ar-H), 7.622-7.585 (1H, t, J = 8.0, Ar-H), 7.483-7.461 (1H, t, J = 8.8, Ar-H), 7.217-7.198 (2H, m, Ar-H), 5.192 (2H, s, -N-CH 2 ), 4.211-4.158 (2H, q, J = 7.2, -O-CH 2 ), 2.505 (3H, s, -CH 3 ), 1.249-1.214 (3H, t, J = 7.2, -CH 3 ); 13 C NMR (100 MHz, DMSO-d 6 ): δ ppm: 168.602, 153.88, 152.217, 148.131, 145.138, 141.430, 137.031, 130.101, 128.833, 128.178, 127.482, 127.271, 125.388, 124.674, 123.720, 122.755, 122.242 , 120.985, 112.485, 109.656, 109.539, 108.235, 61.201, 44.431, 14.026, 9.961. ESIMS found: m/z 452.6 [MH] - , 454.6 [M+H] + , 476.5 [M+Na] + ; Rf = 0.50 (n-Hex:EA=1:2)
<N-아실히드라존 유도체-표지자 접합체의 제조><Preparation of N-acylhydrazone derivative-labeler conjugate>
위 실시예 화합물들을 기초로 N-아실히드라존 유도체-표지자 접합체를 제조하였다. 하기 실시예 29 및 실시예 30의 제조 방법은 p-NCS-벤젠을 매개로 접합되는 경우와 링커 없이 직접 접합되는 경우에 관한 N-아실히드라존 유도체-표지자 접합체의 제조의 일예시를 나타낸다. 아래 예시를 참고하여 위 N-아실히드라존 유도체를 이용하여 통상의 기술자가 상업적으로 이용가능한 화합물(방사성 동위원소의 리간드 또는 형광체 등)과 함께 적절한 합성 조건에 따라 합성을 진행할 수 있다.An N-acylhydrazone derivative-labeler conjugate was prepared based on the compounds of Examples above. The preparation methods of Examples 29 and 30 below show an example of the preparation of the N-acylhydrazone derivative-labeler conjugate in the case of conjugation via p-NCS-benzene and the case of direct conjugation without a linker. Referring to the examples below, a person skilled in the art can proceed with synthesis according to appropriate synthesis conditions using the above N-acylhydrazone derivatives together with commercially available compounds (radioactive isotope ligands or fluorescent substances, etc.).
실시예 29: N-아실히드라존 유도체-리간드(DFO) 접합체의 제조Example 29: Preparation of N-acylhydrazone derivative-ligand (DFO) conjugate
Figure PCTKR2021007223-appb-img-000113
Figure PCTKR2021007223-appb-img-000113
단계 1: t-부틸 (2-(2-히드록시에톡시)에틸)카바메이트의 제조Step 1: Preparation of t-butyl (2-(2-hydroxyethoxy)ethyl)carbamate
Figure PCTKR2021007223-appb-img-000114
Figure PCTKR2021007223-appb-img-000114
2-(2-아미노에톡시)에탄올(1 g, 9.5 mmol, 1 eq)의 수용액(10 mL)에 KOH(0.58 g, 10.4 mmol, 1.1 eq)를 가하였다. 수득한 용액을 0℃에서 교반하고 디-t-부틸 디카보네이트(2.4 mL, 10.4 mmol, 1.1 eq)의 1,4-디옥산 용액(5 mL)을 적가하였다. 1시간 후, 수득한 용액을 실온으로 승온하고 4시간 더 교반하였다. 반응 혼합물을 CH2Cl2 (50 mL)로 추출하고, 유기층을 Na2SO4로 건조하고 증발시켰다. 잔사를 크로마토그래피(EA:n-Hex=8:2)로 정제하여 표제 화합물 1.84 g(수율 94%, 무색 액체)을 얻었다.To an aqueous solution (10 mL) of 2-(2-aminoethoxy)ethanol (1 g, 9.5 mmol, 1 eq) was added KOH (0.58 g, 10.4 mmol, 1.1 eq). The obtained solution was stirred at 0° C. and a 1,4-dioxane solution (5 mL) of di-t-butyl dicarbonate (2.4 mL, 10.4 mmol, 1.1 eq) was added dropwise. After 1 hour, the obtained solution was heated to room temperature and stirred for another 4 hours. The reaction mixture was extracted with CH 2 Cl 2 (50 mL), the organic layer was dried over Na 2 SO 4 and evaporated. The residue was purified by chromatography (EA:n-Hex=8:2) to obtain 1.84 g (yield 94%, colorless liquid) of the title compound.
1H NMR (500 MHz, CDCl3): δ 3.76 - 3.71 (m, 2H), 3.59 - 3.51 (m, 4H), 3.32 (t, J = 5.1 Hz, 2 H), 1.44 (s, 9H); 13C NMR (100 MHz, CDCl3): δ 156.1, 72.1, 70.1, 61.4, 28.2. MS (MALDI-TOF): m/z 228 [M+Na]+. 1 H NMR (500 MHz, CDCl 3 ): δ 3.76 - 3.71 (m, 2H), 3.59 - 3.51 (m, 4H), 3.32 (t, J = 5.1 Hz, 2 H), 1.44 (s, 9H); 13 C NMR (100 MHz, CDCl 3 ): δ 156.1, 72.1, 70.1, 61.4, 28.2. MS (MALDI-TOF): m/z 228 [M+Na] + .
단계 2: t-부틸 (2-(2-브로모에톡시)에틸)카바메이트의 제조Step 2: Preparation of t-butyl (2-(2-bromoethoxy)ethyl)carbamate
Figure PCTKR2021007223-appb-img-000115
Figure PCTKR2021007223-appb-img-000115
t-부틸 (2-(2-히드록시에톡시)에틸)카바메이트(1 g, 4.8 mmol, 1 eq)의 THF 용액(20 mL)에 트리페닐포스핀(1.28 g, 4.8 mmol, 1 eq) 및 CBr4 (1.62 g, 4.8 mmol, 1 eq)를 차례로 가하고 실온에서 2시간 교반하였다. 반응이 종료된 후, 백색 고체를 여과하고 차가운 THF로 세척하였다. 여액을 농축하고 실리카겔 컬럼 크로마토그래피(EA:n-Hex=2:8)로 정제하여 표제 화합물 0.96 g(수율 74%, 무색 액체)을 얻었다.Triphenylphosphine (1.28 g, 4.8 mmol, 1 eq) in THF solution (20 mL) of t-butyl (2- (2-hydroxyethoxy) ethyl) carbamate (1 g, 4.8 mmol, 1 eq) and CBr 4 (1.62 g, 4.8 mmol, 1 eq) were sequentially added, followed by stirring at room temperature for 2 hours. After the reaction was completed, the white solid was filtered and washed with cold THF. The filtrate was concentrated and purified by silica gel column chromatography (EA:n-Hex=2:8) to obtain 0.96 g (yield 74%, colorless liquid) of the title compound.
1H NMR (500 MHz, CDCl3): δ 4.92 (brs, 1H), 3.77 (t, J = 6.0 Hz, 2 H), 3.56 (t, J = 5.1 Hz, 2H), 3.46 (t, J = 6.0 Hz, 2H), 3.37 - 3.29 (m, 2H), 1.44 (s, 9H). 13C NMR (100 MHz, CDCl3): δ 155.9, 79.2, 70.7, 70.0, 40.2, 30.3, 28.3. MS (MALDI-TOF): m/z 307 [M+K]+. 1 H NMR (500 MHz, CDCl 3 ): δ 4.92 (brs, 1H), 3.77 (t, J = 6.0 Hz, 2 H), 3.56 (t, J = 5.1 Hz, 2H), 3.46 (t, J = 6.0 Hz, 2H), 3.37 - 3.29 (m, 2H), 1.44 (s, 9H). 13 C NMR (100 MHz, CDCl 3 ): δ 155.9, 79.2, 70.7, 70.0, 40.2, 30.3, 28.3. MS (MALDI-TOF): m/z 307 [M+K] + .
단계 3: t-부틸 (2-(2-(2-클로로-3-포밀-1H-인돌-1-일)에톡시)에틸)카바메이트의 제조Step 3: Preparation of t-butyl (2-(2-(2-chloro-3-formyl-1H-indol-1-yl)ethoxy)ethyl)carbamate
Figure PCTKR2021007223-appb-img-000116
Figure PCTKR2021007223-appb-img-000116
2-클로로-1H-인돌-3-카복스알데히드(0.134 g, 0.74 mmol, 1 eq)를 아세토니트릴(10 mL)에 넣고 식힌 뒤, 질소 분위기 하에서 Cs2CO3(1.21 g, 3.73 mmol, 5 eq)를 가하였다. 반응 혼합물을 10분간 교반한 뒤 t-부틸 (2-(2-브로모에톡시)에틸)카바메이트(0.2 g, 0.74 mmol, 1 eq)을 적가하고, 15시간 환류하였다. 반응이 완료되면 반응 혼합물을 농축하여 아세토니트릴을 제거하고, 물(15 mL)을 가하여 반응을 종료한 뒤, EA(15 mL)로 2회 추출하였다. 유기 추출물을 취합하고 Na2SO4,로 건조한 뒤 진공 하에서 농축하였다. 조(crude) 생성물을 실리카겔 컬럼 크로마토그래피(EA:Hex=3:7)에 의해 정제하여 표제 화합물 210 mg(수율 77%, 밝은 갈색 액체)를 얻었다.2-Chloro-1H-indole-3-carboxaldehyde (0.134 g, 0.74 mmol, 1 eq) was added to acetonitrile (10 mL), cooled, and Cs 2 CO 3 (1.21 g, 3.73 mmol, 5 eq) was added. After stirring the reaction mixture for 10 minutes, t-butyl (2-(2-bromoethoxy)ethyl)carbamate (0.2 g, 0.74 mmol, 1 eq) was added dropwise, and refluxed for 15 hours. When the reaction was completed, the reaction mixture was concentrated to remove acetonitrile, water (15 mL) was added to terminate the reaction, and then extracted twice with EA (15 mL). The organic extracts were combined , dried over Na 2 SO 4 , and concentrated under vacuum. The crude product was purified by silica gel column chromatography (EA:Hex=3:7) to give 210 mg (yield 77%, light brown liquid) of the title compound.
1H NMR (500 MHz, CDCl3): δ 10.13 (s, 1H), 8.33 - 8.25 (m, 1H), 7.40 - 7.36 (m, 1H), 7.35 - 7.29 (m, 2H), 4.43 (t, J = 5.4 Hz, 2H), 3.79 (t, J = 5.4 Hz, 2H), 3.42 (t, J = 5.1 Hz, 2H), 3.24 - 3.14 (m, 2H), 1.40 (s, 9H). 13C NMR (100 MHz, CDCl3): δ 184.0, 155.7, 136.3, 135.8, 124.3, 124.0, 123.4, 121.3, 113.1, 113.1, 109.9, 79.2, 70.3, 68.8, 43.8, 40.1, 28.3. MS (MALDI-TOF): m/z 389 [M+Na]+. 1 H NMR (500 MHz, CDCl 3 ): δ 10.13 (s, 1H), 8.33 - 8.25 (m, 1H), 7.40 - 7.36 (m, 1H), 7.35 - 7.29 (m, 2H), 4.43 (t, J = 5.4 Hz, 2H), 3.79 (t, J = 5.4 Hz, 2H), 3.42 (t, J = 5.1 Hz, 2H), 3.24 - 3.14 (m, 2H), 1.40 (s, 9H). 13 C NMR (100 MHz, CDCl 3 ): δ 184.0, 155.7, 136.3, 135.8, 124.3, 124.0, 123.4, 121.3, 113.1, 113.1, 109.9, 79.2, 70.3, 68.8, 43.8, 40.1, 28.3. MS (MALDI-TOF): m/z 389 [M+Na] + .
단계 4: 에틸 5-메틸벤조퓨란-2-카복실레이트Step 4: Ethyl 5-methylbenzofuran-2-carboxylate
Figure PCTKR2021007223-appb-img-000117
Figure PCTKR2021007223-appb-img-000117
5-메틸살리실알데히드(1.40 g, 10.3 mmol)의 DMF(30 mL) 용액에 Cs2CO3 (10.07 g, 30.9 mmol) 및 에틸 브로모아세테이트(1.38 mL, 12.4 mmol)를 넣고 70 ℃에서 15시간 반응시켰다. 반응 용액을 물로 희석한 후, EA로 추출하고, 유기층을 MgSO4로 건조 및 여과하고 감압 증류하였다. 잔사를 컬럼 크로마토그래피(n-Hex:EA=10:1)로 분리하여 표제 화합물 1.28 g (수율 61%, 무색 액체)을 얻었다. Cs 2 CO 3 (10.07 g, 30.9 mmol) and ethyl bromoacetate (1.38 mL, 12.4 mmol) were added to a DMF (30 mL) solution of 5-methylsalicylaldehyde (1.40 g, 10.3 mmol), and 15 at 70 ° C. time was reacted. The reaction solution was diluted with water, extracted with EA, and the organic layer was dried over MgSO 4 , filtered, and distilled under reduced pressure. The residue was separated by column chromatography (n-Hex:EA=10:1) to obtain 1.28 g (yield 61%, colorless liquid) of the title compound.
1H NMR (400 MHz, CDCl3): δ 7.49-7.46 (m, 3H), 7.26 (m, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.46 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H); 13C NMR (100 MHz, CDCl3): δ 159.85, 154.37, 145.92, 133.51, 129.27, 127.22, 122.45, 113.74, 112.02, 61.61, 21.45, 14.51; mp 38-39℃ 1 H NMR (400 MHz, CDCl 3 ): δ 7.49-7.46 (m, 3H), 7.26 (m, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.46 (s, 3H), 1.44 (t) , J = 7.2 Hz, 3H); 13 C NMR (100 MHz, CDCl 3 ): δ 159.85, 154.37, 145.92, 133.51, 129.27, 127.22, 122.45, 113.74, 112.02, 61.61, 21.45, 14.51; mp 38-39℃
단계 5: 5-메틸벤조퓨란-2-카보히드라지드Step 5: 5-Methylbenzofuran-2-carbohydrazide
Figure PCTKR2021007223-appb-img-000118
Figure PCTKR2021007223-appb-img-000118
에틸 5-메틸벤조퓨란-2-카복실레이트(2.04 g, 10.0 mmol)를 EtOH(30 mL)에 녹이고 여기에 히드라진 모노히드레이트(1.50 g, 30.0 mmol)를 넣어 24시간 환류시켰다. 반응 용액을 감압증류하고 얻어진 고체를 물로 세척한 다음, 건조하여 표제 화합물 1.72 g (수율 90%, 흰색 고체)을 얻었다. Ethyl 5-methylbenzofuran-2-carboxylate (2.04 g, 10.0 mmol) was dissolved in EtOH (30 mL), hydrazine monohydrate (1.50 g, 30.0 mmol) was added thereto, and refluxed for 24 hours. The reaction solution was distilled under reduced pressure, and the resulting solid was washed with water and dried to obtain 1.72 g of the title compound (yield 90%, white solid).
1H NMR (400 MHz, DMSO-d6): δ 9.97 (s, 1H), 7.52-7.49 (m, 2H), 7.43 (s, 1H), 7.25 (dd, J = 8.4, 1.6 Hz, 1H), 4.55 (br s, 2H), 2.40 (s, 3H); 13C NMR (100 MHz, DMSO-d6): δ 157.92, 152.64, 148.48, 132.68, 127.84, 127.11, 122.04, 111.27, 108.47, 20.81; MS (MALDI-TOF): m/z 213 [M+Na]+; mp 159℃ 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.97 (s, 1H), 7.52-7.49 (m, 2H), 7.43 (s, 1H), 7.25 (dd, J = 8.4, 1.6 Hz, 1H) , 4.55 (br s, 2H), 2.40 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ): δ 157.92, 152.64, 148.48, 132.68, 127.84, 127.11, 122.04, 111.27, 108.47, 20.81; MS (MALDI-TOF): m/z 213 [M+Na] + ; mp 159℃
단계 6: t-부틸 (E)-(2-(2-(2-클로로-3-((2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸)-1H-인돌-1-일)에톡시)에틸)카바메이트의 제조Step 6: t-Butyl (E)-(2-(2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl)-1H-indole- Preparation of 1-yl) ethoxy) ethyl) carbamate
Figure PCTKR2021007223-appb-img-000119
Figure PCTKR2021007223-appb-img-000119
t-부틸 (2-(2-(2-클로로-3-포밀-1H-인돌-1-일)에톡시)에틸)카바메이트(0.14 g, 0.38 mmol, 1 eq) 및 5-메틸벤조퓨란-2-카보히드라지드(74 mg, 0.38 mmol, 1 eq)를 n-프로판올(10 mL)에 넣고 아세트산을 촉매량(2 방울) 가한 뒤, 가열하여 6시간 환류시켰다. 반응이 종료된 후(TLC 관찰), 반응 혼합물을 실온으로 식히고 회전증발기에서 용매를 제거하였다. 수득한 고상의 조 생성물에 에탄올 15 mL를 0℃에서 가하고 10분간 교반한 뒤, 여과하고 차가운 에탄올(5 mL)로 세척하였다. 마지막으로 여액을 진공 하에 건조하여 표제 화합물 180 mg(수율 87%, 밝은 황색 고체)을 얻었다. t-Butyl (2-(2-(2-chloro-3-formyl-1H-indol-1-yl)ethoxy)ethyl)carbamate (0.14 g, 0.38 mmol, 1 eq) and 5-methylbenzofuran- 2-Carbohydrazide (74 mg, 0.38 mmol, 1 eq) was added to n-propanol (10 mL), acetic acid was added in a catalytic amount (2 drops), and then heated to reflux for 6 hours. After completion of the reaction (TLC observation), the reaction mixture was cooled to room temperature and the solvent was removed on a rotary evaporator. To the obtained solid crude product, 15 mL of ethanol was added at 0° C., stirred for 10 minutes, filtered, and washed with cold ethanol (5 mL). Finally, the filtrate was dried under vacuum to give 180 mg (yield 87%, light yellow solid) of the title compound.
1H NMR (500 MHz, DMSO-d6): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.31 (d, J = 7.7 Hz, 1H), 7.68 - 7.51 (m, 4H), 7.33 - 7.23 (m, 3H), 6.71 - 6.58 (m, 1H), 4.45 (t, J = 5.3 Hz, 2H), 3.72 (t, J = 5.3, 2H), 3.35 (t, J = 5.4 Hz, 2H), 2.99 (q, J = 5.6 Hz, 2H), 2.43 (s, 3H), 1.35 (s, 9H). 13C NMR (100 MHz, DMSO-d6): δ 155.5, 154.2, 152.8, 148.3, 135.7, 132.9, 129.1, 127.1, 123.3, 111.3, 107.5, 77.5, 69.1, 68.1, 43.4, 28.1, 20.8, 13.5. MS (MALDI-TOF): m/z 561 [M+Na]+. mp 93-95℃ 1 H NMR (500 MHz, DMSO-d 6 ): δ 12.01 (s, 1H), 8.75 (s, 1H), 8.31 (d, J = 7.7 Hz, 1H), 7.68 - 7.51 (m, 4H), 7.33 - 7.23 (m, 3H), 6.71 - 6.58 (m, 1H), 4.45 (t, J = 5.3 Hz, 2H), 3.72 (t, J = 5.3, 2H), 3.35 (t, J = 5.4 Hz, 2H) ), 2.99 (q, J = 5.6 Hz, 2H), 2.43 (s, 3H), 1.35 (s, 9H). 13 C NMR (100 MHz, DMSO-d 6 ): δ 155.5, 154.2, 152.8, 148.3, 135.7, 132.9, 129.1, 127.1, 123.3, 111.3, 107.5, 77.5, 69.1, 68.1, 43.4, 28.1, 20.8, 13.5. MS (MALDI-TOF): m/z 561 [M+Na] + . mp 93-95℃
단계 7: (E)-N'-((1-(2-(2-아미노에톡시)에틸)-2-클로로-1H-인돌-3-일)메틸렌)-5-메틸벤조퓨란-2-카보히드라지드 히드로클로라이드의 제조Step 7: (E)-N'-((1-(2-(2-aminoethoxy)ethyl)-2-chloro-1H-indol-3-yl)methylene)-5-methylbenzofuran-2- Preparation of Carbohydrazide Hydrochloride
Figure PCTKR2021007223-appb-img-000120
Figure PCTKR2021007223-appb-img-000120
t-부틸 (E)-(2-(2-(2-클로로-3-((2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸)-1H-인돌-1-일)에톡시)에틸)카바메이트(90 mg)을 건조된 아세토니트릴(5 mL)에 용해하고 2 M HCl의 디에틸에테르 용액(2 mL)을 가한 뒤 실온에서 2시간 교반하였다. 출발 물질이 사라진 것을 TLC로 확인한 후, 반응 혼합물을 농축하여 아세토니트릴을 제거하고, 20% CH2Cl2의 디에틸에테르 용액(10 mL)으로 2회 세척하였다. 수득물을 높은 진공으로 건조하여 표제 화합물 80 mg(수율 95%, 밝은 황색 고체)를 얻었다. t-Butyl (E)-(2-(2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl )Ethoxy)ethyl)carbamate (90 mg) was dissolved in dried acetonitrile (5 mL), 2 M HCl in diethyl ether solution (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After TLC confirmed that the starting material had disappeared, the reaction mixture was concentrated to remove acetonitrile, and washed twice with 20% CH 2 Cl 2 diethyl ether solution (10 mL). The obtained product was dried under high vacuum to give 80 mg (yield 95%, light yellow solid) of the title compound.
1H NMR (500 MHz, DMSO-d6): δ 12.11 (s, 1H), 8.79 (s, 1H), 8.32 (d, J = 7.7 Hz, 1H), 7.97 (brs, 3H), 7.72 - 7.55 (m, 4H), 7.36 - 7.23 (m, 3H), 4.50 (t, J = 5.4 Hz, 2H), 3.79 (t, J = 5.3 Hz, 2H), 3.59 (t, J = 5.3 Hz, 2 H), 2.89 (q, J = 5.4 Hz, 2H), 2.43 (s, 3H). 13C NMR (100 MHz, DMSO-d6): δ 154.2, 152.8, 148.3, 135.7, 132.9, 129.0, 127.1, 123.3, 122.2, 111.3, 110.0, 107.6, 68.4, 66.5, 43.2, 38.2, 20.8. MS (MALDI-TOF): m/z 439 [M+H]+. mp 214-216℃ 1 H NMR (500 MHz, DMSO-d 6 ): δ 12.11 (s, 1H), 8.79 (s, 1H), 8.32 (d, J = 7.7 Hz, 1H), 7.97 (brs, 3H), 7.72 - 7.55 (m, 4H), 7.36 - 7.23 (m, 3H), 4.50 (t, J = 5.4 Hz, 2H), 3.79 (t, J = 5.3 Hz, 2H), 3.59 (t, J = 5.3 Hz, 2H) ), 2.89 (q, J = 5.4 Hz, 2H), 2.43 (s, 3H). 13 C NMR (100 MHz, DMSO-d 6 ): δ 154.2, 152.8, 148.3, 135.7, 132.9, 129.0, 127.1, 123.3, 122.2, 111.3, 110.0, 107.6, 68.4, 66.5, 43.2, 38.2, 20.8. MS (MALDI-TOF): m/z 439 [M+H] + . mp 214-216℃
단계 8: (E)-N1-(5-(3-(4-(3-(2-(2-(2-클로로-3-((2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸)-1H-인돌-1-일)에톡시)에틸)티오우레이도)페닐)티오우레이도)펜틸)-N1-히드록시-N4-(5-(N-히드록시-4-((5-(N-히드록시아세트아미도)펜틸)아미노)-4-옥소부탄아미도)펜틸)석신아미드의 제조 Step 8: (E)-N 1 -(5-(3-(4-(3-(2-(2-(2-chloro-3-((2-(5-methylbenzofuran-2-carbonyl) )hydrazinylidene)methyl)-1H-indol-1-yl)ethoxy)ethyl)thioureido)phenyl)thioureido)pentyl)-N 1 -hydroxy-N 4 -(5-(N) Preparation of -hydroxy-4-((5-(N-hydroxyacetamido)pentyl)amino)-4-oxobutanamido)pentyl)succinamide
Figure PCTKR2021007223-appb-img-000121
Figure PCTKR2021007223-appb-img-000121
(E)-N'-((1-(2-(2-아미노에톡시)에틸)-2-클로로-1H-인돌-3-일)메틸렌)-5-메틸벤조퓨란-2-카보히드라지드 히드로클로라이드(1.3 mg, 0.0027 mmol, 1 eq)를 건조된 DMSO에 넣고 디이소프로필에틸아민(0.7 mg, 0.0053 mmol, 2 eq)을 가한 뒤 5분간 교반하였다. 반응 혼합물에 디페록사민-p-SCN(2 mg, 0.0027 mmol, 1 eq)을 가하고 실온에서 2시간 교반하였다. 조 생성물을 역상 고성능 액체 크로마토그래피(RP-HPLC)에 의해 정제하여 표제 화합물 2.3 mg(수율: 72%, 흰색 고체)을 얻었다.(E)-N'-((1-(2-(2-aminoethoxy)ethyl)-2-chloro-1H-indol-3-yl)methylene)-5-methylbenzofuran-2-carbohydrazide Hydrochloride (1.3 mg, 0.0027 mmol, 1 eq) was placed in dried DMSO, diisopropylethylamine (0.7 mg, 0.0053 mmol, 2 eq) was added thereto, followed by stirring for 5 minutes. To the reaction mixture was added deferoxamine-p-SCN (2 mg, 0.0027 mmol, 1 eq) and stirred at room temperature for 2 hours. The crude product was purified by reverse phase high performance liquid chromatography (RP-HPLC) to obtain 2.3 mg (yield: 72%, white solid) of the title compound.
MS (MALDI-TOF): m/z 1193 [M+2H]+ and 1214[M+Na] + MS (MALDI-TOF): m/z 1193 [M+2H] + and 1214[M+Na] +
또한 상기 실시예 29의 화합물의 HPLC 결과를 하기 표 1에 정리하였다.In addition, the HPLC results of the compound of Example 29 are summarized in Table 1 below.
머무름 시간
[min]dwell time
[min] 		면적
[mVs]area
[mVs] 		높이
[mV]Height
[mV] 		면적
[%]area
[%] 		높이
[%]Height
[%] 		W05
[min]W05
[min]
1One 6.5676.567 2833.7202833.720 10.08510.085 1.61.6 0.20.2 7.307.30
22 8.5178.517 466.223466.223 14.50314.503 0.30.3 0.30.3 0.600.60
33 9.1839.183 1089.8891089.889 12.97412.974 0.60.6 0.30.3 0.600.60
44 17.05017.050 3.6333.633 0.1200.120 0.00.0 0.00.0 0.300.30
55 17.6017.60 2.5082.508 0.1570.157 0.00.0 0.00.0 0.280.28
66 18.63318.633 12.09012.090 0.2810.281 0.00.0 0.00.0 0.750.75
77 19.76719.767 567.587567.587 8.9398.939 0.30.3 0.20.2 0.830.83
88 23.43323.433 3239.0403239.040 98.43198.431 1.81.8 2.02.0 0.500.50
99 27.41727.417 172316.799172316.799 4655.8564655.856 94.894.8 96.696.6 0.530.53
1010 29.13329.133 1111.1271111.127 17.65317.653 0.60.6 0.40.4 0.980.98
1111 35.38335.383 39.20039.200 0.8910.891 0.00.0 0.00.0 0.720.72
TotalTotal 181681.816181681.816 4817.8904817.890 100.0100.0 100.0100.0
실시예 30: N-아실히드라존 유도체-형광체(Cy5) 접합체의 제조Example 30: Preparation of N-acylhydrazone derivative-phosphor (Cy5) conjugate
Figure PCTKR2021007223-appb-img-000122
Figure PCTKR2021007223-appb-img-000122
실시예 29의 단계 7에서 얻은 (E)-N'-((1-(2-(2-아미노에톡시)에틸)-2-클로로-1H-인돌-3-일)메틸렌)-5-메틸벤조퓨란-2-카보히드라지드 히드로클로라이드(3.0 mg, 0.0065 mmol, 1 eq)를 건조된 DMF에 넣고 디이소프로필에틸아민(1.7 mg, 0.013 mmol, 2 eq)을 가한 뒤 5분간 교반하였다. 반응 혼합물에 시아닌 5 NHS 에스테르(4 mg, 0.0065 mmol, 1 eq)를 가하고 실온에서 2시간 교반하였다. 조 생성물을 역상 고성능 액체 크로마토그래피(RP-HPLC)에 의해 정제하여 표제 화합물 5.8 mg(수율: 99%, 자주색 고체)을 얻었다.(E)-N'-((1-(2-(2-aminoethoxy)ethyl)-2-chloro-1H-indol-3-yl)methylene)-5-methyl obtained in step 7 of Example 29 Benzofuran-2-carbohydrazide hydrochloride (3.0 mg, 0.0065 mmol, 1 eq) was placed in dried DMF, diisopropylethylamine (1.7 mg, 0.013 mmol, 2 eq) was added thereto, followed by stirring for 5 minutes. Cyanine 5 NHS ester (4 mg, 0.0065 mmol, 1 eq) was added to the reaction mixture and stirred at room temperature for 2 hours. The crude product was purified by reverse phase high performance liquid chromatography (RP-HPLC) to give 5.8 mg (yield: 99%, purple solid) of the title compound.
MS (MALDI-TOF): m/z 905 [M+H]. MS (MALDI-TOF): m/z 905 [M+H].
또한 상기 실시예 30의 화합물의 HPLC 결과를 하기 표 2에 정리하였다.In addition, the HPLC results of the compound of Example 30 are summarized in Table 2 below.
머무름 시간
[min]dwell time
[min] 		면적
[mVs]area
[mVs] 		높이
[mV]Height
[mV] 		면적
[%]area
[%] 		높이
[%]Height
[%] 		W05
[min]W05
[min]
1One 8.3678.367 822.763822.763 32.43432.434 0.40.4 0.70.7 0.470.47
22 9.0009.000 2032.6952032.695 29.07429.074 1.01.0 0.60.6 1.081.08
33 13.78313.783 13.87813.878 0.6600.660 0.00.0 0.00.0 0.350.35
44 15.85015.850 38.86638.866 0.8430.843 0.00.0 0.00.0 0.270.27
55 17.35017.350 22.21722.217 0.8380.838 0.00.0 0.00.0 0.450.45
66 18.15018.150 30.22230.222 1.1901.190 0.00.0 0.00.0 0.350.35
77 19.58319.583 210.402210.402 8.6738.673 0.10.1 0.20.2 0.450.45
88 20.18320.183 274.940274.940 4.5434.543 0.10.1 0.10.1 0.880.88
99 22.65022.650 54.65754.657 1.2491.249 0.00.0 0.00.0 0.570.57
1010 23.88323.883 49.01849.018 1.9061.906 0.00.0 0.00.0 0.480.48
1111 25.03325.033 236.965236.965 5.2275.227 0.10.1 0.10.1 0.730.73
1212 25.48325.483 127.794127.794 4.0764.076 0.10.1 0.10.1 0.450.45
1313 27.25027.250 157.269157.269 2.8202.820 0.10.1 0.10.1 1.151.15
1414 28.41728.417 61.84661.846 1.6851.685 0.00.0 0.00.0 0.680.68
1515 29.28329.283 11.07111.071 0.4430.443 0.00.0 0.00.0 0.450.45
1616 30.88330.883 195.864195.864 3.7443.744 0.10.1 0.10.1 0.900.90
1717 32.08332.083 2978.2182978.218 69.06369.063 1.51.5 1.41.4 0.650.65
1818 34.60034.600 194972.897194972.897 4750.9964750.996 96.396.3 96.696.6 0.520.52
1919 39.23339.233 113.099113.099 1.0911.091 0.10.1 0.00.0 1.901.90
TotalTotal 202404.681202404.681 4920.5524920.552 100.0100.0 100.0100.0
상기 실시예의 화합물들에 대해 아래의 실험을 수행하였다.The following experiments were performed on the compounds of the above Examples.
실험 기기laboratory instrument
1. 액상 크로마토그래피(LC)1. Liquid Chromatography (LC)
- HPLC 시스템: Agilent 1260 시리즈(on-line degasser, binary pump, thermostatted well-plate autosampler 및 column compartment 장착)- HPLC system: Agilent 1260 series with on-line degasser, binary pump, thermostatted well-plate autosampler and column compartment
- 가드 컬럼: henomenex Security Guard C18 (2.0 x 4 mm)- Guard column: henomenex Security Guard C18 (2.0 x 4 mm)
- 컬럼: Waters Xterra MS C18 (2.1 x 50 mm, 5 μm)- Column: Waters Xterra MS C18 (2.1 x 50 mm, 5 μm)
- 이동상: 95% A (0.1% 포름산 수용액) / 5% B (0.1% 포름산 아세토니트릴 용액)에서 5% A/ 95% B까지의 선형 기울기- Mobile phase: linear gradient from 95% A (0.1% aqueous formic acid solution) / 5% B (0.1% acetonitrile formic acid solution) to 5% A / 95% B
- 유속: 0.4 mL/min- Flow rate: 0.4 mL/min
2. 질량 분석(MS)2. Mass Spectrometry (MS)
- MS 시스템: 3200 Q TRAP LC/MS/MS 시스템- MS system: 3200 Q TRAP LC/MS/MS system
- 이온 소스: Turbo VTM 이온 스프레이- Ion Source: Turbo V TM Ion Spray
- 극성: 양성- Polarity: positive
- MRM: 실시예 4의 화합물 - m/z 424>159 (Ret. Time: 3.8 min), 카르바마제핀 (IS) - m/z 237>194 (Ret. Time: 3.2 min)- MRM: the compound of Example 4 - m/z 424>159 (Ret. Time: 3.8 min), carbamazepine (IS) - m/z 237>194 (Ret. Time: 3.2 min)
실험예 1: 항증식 활성 시험Experimental Example 1: Anti-proliferative activity test
1.1. HeLa 세포주에서의 항증식 활성 시험1.1. Anti-proliferative activity test in HeLa cell line
본 발명에 따른 실시예 화합물의 항증식 활성에 대해 시험하기 위하여 하기의 실험을 수행하였다.The following experiment was performed to test the antiproliferative activity of the example compounds according to the present invention.
인간 자궁경부암 세포주인 HeLa 세포(ATCC, 미국)를 96-웰 플레이트에 3 x 103 세포/웰로 뿌린 후, 본 발명에 따른 실시예의 화합물로 세포를 처리하고 2일 동안 성장시켰다. 이후 MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) 시약을 10 μL/웰로 첨가하였다. 2시간 후, "OD 450"에서 흡광도를 측정하고 PrismTM 6 프로그램을 사용하여 통계적 수치를 얻었다. 아래 표 3에 나타낸 데이터는 분석을 2번 반복하여 수행한 결과의 평균값이다.HeLa cells (ATCC, USA), a human cervical cancer cell line, were seeded in a 96-well plate at 3 x 10 3 cells/well, and then the cells were treated with the compounds of Examples according to the present invention and grown for 2 days. Then, MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reagent was added at 10 μL/well. After 2 hours, absorbance was measured at "OD 450" and statistical values were obtained using Prism TM 6 program. The data shown in Table 3 below are average values of the results of repeating the analysis twice.
처리물질material to be treated IC50 (nM)IC 50 (nM) 화합물계열compound series
대조군control -- DMSODMSO
실시예 1Example 1 3878.83878.8 벤조퓨란benzofuran
실시예 2Example 2 892.3892.3 벤조퓨란benzofuran
실시예 3Example 3 243.4243.4 벤조퓨란benzofuran
실시예 4Example 4 180.0180.0 벤조퓨란benzofuran
실시예 5Example 5 262.8262.8 벤조퓨란benzofuran
실시예 6Example 6 1216.51216.5 벤조퓨란benzofuran
실시예 7Example 7 232.8232.8 벤조퓨란benzofuran
실시예 8Example 8 82.082.0 벤조퓨란benzofuran
실시예 9Example 9 232.1232.1 벤조퓨란benzofuran
실시예 10Example 10 221.3221.3 벤조퓨란benzofuran
실시예 11Example 11 242.6242.6 벤조퓨란benzofuran
실시예 12Example 12 618.9618.9 벤조퓨란benzofuran
실시예 13Example 13 337.0337.0 벤조퓨란benzofuran
실시예 14Example 14 261.7261.7 벤조퓨란benzofuran
실시예 15Example 15 123.2123.2 벤조퓨란benzofuran
실시예 16Example 16 455.2455.2 벤조퓨란benzofuran
실시예 18Example 18 563 (±37)563 (±37) 벤조퓨란benzofuran
실시예 19Example 19 4524.34524.3 벤조퓨란benzofuran
실시예 20Example 20 500 (±45)500 (±45) 벤조퓨란benzofuran
실시예 21Example 21 199 (±66)199 (±66) 벤조티오펜benzothiophene
실시예 22Example 22 1001 (±171)1001 (±171) 페닐퓨란Phenylfuran
실시예 23Example 23 200 (±113)200 (±113) 페닐티오펜phenylthiophene
실시예 24Example 24 400.5 (±198.5)400.5 (±198.5) 페닐티오펜phenylthiophene
실시예 25Example 25 432 (±241.68)432 (±241.68) 페닐피라졸Phenylpyrazole
실시예 26Example 26 32.8(±0.5)32.8 (±0.5) 나프토퓨란Naphthofuran
실시예 27Example 27 332 (±7.82)332 (±7.82) 나프토퓨란Naphthofuran
실시예 28Example 28 562.3562.3 나프토퓨란Naphthofuran
상기 표 3에 나타난 바와 같이, 본 발명에 따른 실시예의 화합물은 HeLa 세포주(자궁경부암 세포주)에서의 항증식 활성이 우수함을 알 수 있다.As shown in Table 3, it can be seen that the compounds of Examples according to the present invention have excellent anti-proliferative activity in the HeLa cell line (cervical cancer cell line).
1.2. 다른 암세포주에서의 항증식 활성 시험1.2. Anti-proliferative activity test in different cancer cell lines
본 발명에 따른 실시예 화합물이 HeLa 세포주(자궁경부암 세포주)뿐 아니라 다른 암세포에도 항증식 활성을 보이는지 여부를 시험하기 위해 하기의 실험을 수행하였다.The following experiment was performed to test whether the Example compound according to the present invention exhibits anti-proliferative activity not only in the HeLa cell line (cervical cancer cell line) but also in other cancer cells.
구체적으로, 미량정량판(microtiter plate, 1-3x103 세포/웰)에 다양한 암세포주를 배양하고 본 발명에 따른 실시예 4의 화합물로 처리하여 4일간 배양하였다. 세포독성은 상기 실험예 1-1과 동일한 방식의 MTT 분석으로 시험하였고, IC50은 로그 용량 반응 곡선(log-dose response curve)에 의해 얻었다. 하기 표 4에 나타낸 데이터는 분석을 3번 반복하여 수행한 결과의 평균값이다.Specifically, various cancer cell lines were cultured in a microtiter plate (1-3x10 3 cells/well), treated with the compound of Example 4 according to the present invention, and cultured for 4 days. Cytotoxicity was tested by MTT analysis in the same manner as in Experimental Example 1-1, and IC 50 was obtained by a log-dose response curve. The data shown in Table 4 below are average values of the results obtained by repeating the analysis three times.
세포주 유래 조직cell line-derived tissue 세포주cell line 실시예 4 IC50(μM) Example 4 IC 50 (μM)
뼈bone U-2-OSU-2-OS 0.42±0.060.42±0.06
뇌brain A172A172 0.79±0.090.79±0.09
SK-N-MCSK-N-MC 0.98±0.110.98±0.11
U373MGU373MG 0.36±0.010.36±0.01
유방breast HCC1954HCC1954 0.41±0.050.41±0.05
MDA-MB-468MDA-MB-468 0.69±0.030.69±0.03
CNSCNS SNB75SNB75 0.39±0.030.39±0.03
결장
(colon)colon
(colon) 		Colo 205Colo 205 0.36±0.490.36±0.49
HCT 116HCT 116 0.45±0.220.45±0.22
HCT-15HCT-15 0.28±0.070.28±0.07
신장kidney 786-O786-O 0.13±0.050.13±0.05
A498A498 0.17±0.0080.17±0.008
골수
(leukemia)marrow
(leukemia) 		K562K562 0.47±0.0120.47±0.012
MOLT4MOLT4 0.30±0.010.30±0.01
RPMI-8226RPMI-8226 0.52±0.130.52±0.13
간liver Hep3BHep3B 0.19±0.050.19±0.05
A549A549 0.63±0.120.63±0.12
폐lung NCI-H125NCI-H125 0.37±0.20.37±0.2
NCI-H1299NCI-H1299 0.56±0.150.56±0.15
NCI-H226NCI-H226 0.42±0.060.42±0.06
NCI-H460NCI-H460 0.5±0.090.5±0.09
NCI-H522NCI-H522 0.37±0.130.37±0.13
말초 혈액
(peripheral blood)peripheral blood
(peripheral blood) 		CCRF-CEMCCRF-CEM 0.34±0.020.34±0.02
전립선prostate DU145DU145 0.49±0.090.49±0.09
PC3PC3 0.71±0.180.71±0.18
피부skin A431A431 0.22±0.0170.22±0.017
SK-MEL 5SK-MEL 5 0.46±0.140.46±0.14
위stomach SNU 484SNU 484 0.35±0.070.35±0.07
자궁경부cervix HeLa CCL2HeLa CCL2 0.18±0.0040.18±0.004
정상 세포normal cells MEF(마우스)MEF (mouse) 4.01±4.584.01±4.58
HDF(인간)HDF (human) 27.0±7.8927.0±7.89
상기 표 4에 나타난 바와 같이, 본 발명에 따른 실시예 화합물은 다양한 암세포주에서도 항증식 활성이 우수함을 알 수 있다.As shown in Table 4, it can be seen that the example compounds according to the present invention have excellent antiproliferative activity even in various cancer cell lines.
1.3. 다중약물 내성을 보이는 암세포에서의 항증식 활성 시험1.3. Anti-proliferative activity test in cancer cells showing multidrug resistance
본 발명에 따른 실시예 화합물이 다중약물 내성을 보이는 암세포주에도 효과를 보이는지 시험하기 위하여 하기의 실험을 수행하였다.The following experiment was performed to test whether the Example compound according to the present invention has an effect on cancer cell lines showing multi-drug resistance.
구체적으로, K562 및 MCF7(바이오평가센터, 한국생명공학연구원, 한국) 및 이들 각 세포주의 다중약물 내성을 가진 세포주인 K562/ADR과 MCF7/ADR(바이오평가센터, 한국생명공학연구원, 한국)를 미량정량판(microtiter plate, 1-3x103 세포/웰)에 배양하였다. 이후 세포들을 본 발명에 따른 실시예 4의 화합물, 탁솔(taxol), 독소루비신(doxorubicin), 빈블라스틴(vinblastine) 또는 콜히친(colchicine)으로 처리하여 4일간 배양하였다. 세포독성은 상기 실험예 1-1과 동일한 방식의 MTT 분석으로 시험하였고, IC50은 로그 용량 반응 곡선(log-dose response curve)에 의해 얻었다(단위: nM). 하기 표에 나타낸 데이터는 분석을 3번 반복하여 수행한 결과의 평균값이다.Specifically, K562 and MCF7 (Bio Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Korea) and K562/ADR and MCF7/ADR (Bio Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Korea), which are cell lines with multi-drug resistance of each of these cell lines, were tested. It was cultured in a microtiter plate (1-3x10 3 cells/well). Thereafter, the cells were treated with the compound of Example 4 according to the present invention, taxol, doxorubicin, vinblastine or colchicine, and cultured for 4 days. Cytotoxicity was tested by MTT analysis in the same manner as in Experimental Example 1-1, and IC 50 was obtained by a log-dose response curve (unit: nM). The data shown in the table below are the average values of the results obtained by repeating the analysis three times.
다중약물 내성을 보이는 세포주의 저항인자(resistance factor)는 내성이 없는 모세포주(parent)의 IC50에 대한 다중약물 내성 세포주의 IC50의 비율을 의미한다. Resistance factors (resistance factor) of a cell line exhibit multidrug resistance; means the ratio of the IC 50 of the multi-drug-resistant cell line to the IC 50 of the primary (parent) cells with no resistance.
IC50(nM) IC 50 (nM) 실시예 4Example 4 탁솔Taxol 독소루비신doxorubicin 빈블라스틴 vinblastine 콜히친colchicine
K562K562 350350 1.3471.347 2.8482.848 17.1317.13 15.8615.86
K562/ADRK562/ADR 200.8200.8 699.9699.9 21872187 267.8267.8 363.4363.4
저항인자resistance factor 0.570.57 519.6519.6 767.91767.91 15.6315.63 22.9122.91
IC50(nM) IC 50 (nM) 실시예 4Example 4 탁솔Taxol 독소루비신doxorubicin 빈블라스틴vinblastine 콜히친colchicine
MCF7MCF7 822822 2.3452.345 15.3815.38 5.2755.275 2.712.71
MCF7/ADRMCF7/ADR 331.5331.5 10281028 26082608 95.8795.87 92.5492.54
저항인자resistance factor 0.400.40 438.38438.38 169.57169.57 18.1718.17 34.1534.15
상기 표 5 및 6에 나타난 바와 같이, 다중약물 내성 세포주는 기존의 항암제에 대해서 저항인자가 수십 내지 수백 배에 달하는 큰 저항성을 나타내었다. 그러나, 본 발명에 따른 실시예 화합물에 대해서는 저항 인자가 0.40 내지 0.57로 나타나, 본 발명에 따른 실시예 화합물이 기존의 항암제에 비해 다중약물 내성을 보이는 암세포주에 더 강한 세포독성 효과를 나타냄을 알 수 있다. As shown in Tables 5 and 6, the multi-drug-resistant cell line exhibited great resistance, tens to hundreds of times the resistance factor to the existing anticancer drugs. However, for the example compound according to the present invention, the resistance factor was 0.40 to 0.57, indicating that the example compound according to the present invention exhibits a stronger cytotoxic effect on cancer cell lines showing multidrug resistance compared to the existing anticancer drugs. can
실험예 2: 세포주기 진행에 미치는 효과 시험Experimental Example 2: Effect test on cell cycle progression
HeLa CCL2 세포주를 12 웰 플레이트(3x104 세포/웰)에서 배양하고, DMSO 또는 실시예 4의 화합물로 17 시간 동안 처리한 후, 프로피디움 요오드(propidium iodine) 염료를 첨가하여 세포 DNA를 염색하고 FACS를 사용하여 측정하였다. 하기 표에, 분열기에 세포가 모이는 농도와 세포의 수를 퍼센트로 나타내었다The HeLa CCL2 cell line was cultured in a 12-well plate (3x10 4 cells/well), treated with DMSO or the compound of Example 4 for 17 hours, and then propidium iodine dye was added to stain the cell DNA and FACS. was used. In the table below, the cell concentration and the number of cells in the dividing phase are expressed as a percentage.
세포주cell line 효과 μM (%, G2/M기)Effect μM (%, G2/M phase)
HeLa CCL2HeLa CCL2 0.2 (82.36)0.2 (82.36)
상기 표 7에 나타난 바와 같이, 실시예 4의 화합물은 대표적인 암세포주인 HeLa CCL2에 대해 0.2μM에서 IC50을 보이며, 특히 세포에 처리한 후 16시간 이후 80% 이상의 세포가 분열기(mitotic phase)에 멈춰서 있는 것을 관찰함으로써 튜불린의 중합 반응을 억제하는 것으로 기대되어 이를 실험예 3에서 확인하였다.As shown in Table 7, the compound of Example 4 showed an IC 50 at 0.2 μM against HeLa CCL2, a representative cancer cell line, and in particular, more than 80% of the cells stopped in the mitotic phase after 16 hours after treatment with the cells. It was expected to inhibit the polymerization reaction of tubulin by observing that there is, and this was confirmed in Experimental Example 3.
실험예 3: 튜불린 중합에 미치는 효과 시험Experimental Example 3: Effect test on tubulin polymerization
본 발명에 따른 실시예 화합물이 세포내 미세소관에 미치는 효과를 확인하기 위해, HeLa 세포를 DMSO 또는 실시예 4의 화합물(50 nM, 100 nM, 200 nM)로 16시간 동안 처리하였다. In order to confirm the effect of the example compound according to the present invention on intracellular microtubules, HeLa cells were treated with DMSO or the compound of Example 4 (50 nM, 100 nM, 200 nM) for 16 hours.
세포를 고정한 후, 항튜불린 항체(anti-tubulin antibody)와 Alexa FluorTM 488로 염색시키고, Hoechst 33342를 사용하여 세포의 핵을 면역 염색하여 α-튜불린 및 DNA를 시험하였다. 도 1에 나타난 바와 같이, DMSO 대조군과 비교하여 본 발명에 따른 실시예 화합물로 처리하였을 때 농도가 높아질수록 튜불린의 모양이 엉성하고 짧아졌다. 또한 중앙 배열을 이탈한 DNA 형태도 증가하였다.After fixing the cells, they were stained with an anti-tubulin antibody and Alexa Fluor ™ 488, and nuclei of the cells were immunostained using Hoechst 33342 to test α-tubulin and DNA. As shown in FIG. 1 , the shape of tubulin became coarser and shorter as the concentration increased when treated with the example compound according to the present invention compared to the DMSO control group. In addition, the form of DNA deviating from the central alignment also increased.
따라서, 본 발명에 따른 실시예 화합물은 미세소관을 탈중합시키는 제제임을 알 수 있다.Accordingly, it can be seen that the Example compound according to the present invention is an agent for depolymerizing microtubules.
실험예 4: 인체 유래 자궁경부암 세포(HeLa CCL2) 이식 모델에서의 항암 효과 시험 Experimental Example 4: Anticancer effect test in human cervical cancer cell (HeLa CCL2) transplantation model
4.1. 암세포 배양 및 암세포 이식4.1. Cancer cell culture and cancer cell transplantation
액체 질소 속에서 냉동보관 중이던 인체 유래 피부암 세포주 HeLa CCL2를 해동한 후 세포 배양을 실시하였다. 세포의 배양은 CO2 배양기(Forma, USA)에서 온도 37℃ 및 CO2 농도 5%로 맞춰서 적절한 기간 동안 수행되었다.After thawing the human-derived skin cancer cell line HeLa CCL2 that was stored frozen in liquid nitrogen, cell culture was performed. Cell culture was carried out for an appropriate period of time in a CO 2 incubator (Forma, USA) at a temperature of 37° C. and a CO 2 concentration of 5%.
배양 최종일에 모든 암세포를 수거하여 계수하고 무혈청 배양액(serum free media)을 이용하여 세포 농도를 1 x 107 세포/mL로 조절하였다. 이렇게 조절된 세포 배양액을 BALB/C 암컷 누드 마우스(5주령, 나라바이오텍) 당 0.3 mL(3 x 106 세포/마우스)씩 어깨뼈와 흉벽 사이의 액와(axillary) 부위 피하에 주입하였다.On the last day of culture, all cancer cells were collected and counted, and the cell concentration was adjusted to 1 x 10 7 cells/mL using serum free media. 0.3 mL (3 x 10 6 cells/mouse) per BALB/C female nude mouse (5 weeks old, Nara Biotech) was injected subcutaneously into the axillary region between the shoulder blade and the chest wall.
4.2. 시료의 제조 및 투여 방법4.2. Methods of Preparation and Administration of Samples
시험물질로서 본 발명에 따른 실시예 화합물을 사용하고, 음성 대조물질로서 용매(담체)를 사용하였다. Example compounds according to the present invention were used as test substances, and solvents (carriers) were used as negative controls.
화합물들을 투여 직전에 DMAC(디메틸아세트아미드) 20% + Tween 80 5% + 20% HPbCD(2-히드록시프로필-β-시클로덱스트린) 75%의 혼합액에 적절한 농도로 용해한 후 사용하였다. 제조된 물질들은 마우스 20 g당 0.2 mL(10 mL/kg)씩 아래 투여 스케줄로 반복 복강 투여하였다.Just before administration, the compounds were dissolved in a mixture of DMAC (dimethylacetamide) 20% + Tween 80 5% + 20% HPbCD (2-hydroxypropyl-β-cyclodextrin) 75% to an appropriate concentration before use. The prepared substances were repeatedly administered intraperitoneally by 0.2 mL (10 mL/kg) per 20 g of a mouse according to the following dosing schedule.
- 담체, 실시예 4의 화합물, 실시예 8의 화합물 및 실시예 15의 화합물 (25 mg/kg), 0-28일- carrier, the compound of example 4, the compound of example 8 and the compound of example 15 (25 mg/kg), 0-28 days
4.3. 종양크기 변화 확인4.3. Change in tumor size
암세포 이식 후, 평균 종양 크기가 57.0 mm3 도달 시부터 28일째까지 동물 개체별 종양 크기를 버니어캘리퍼스(vernier caliper)를 이용하여 3 방향에 대해 총 11회 측정한 후, 길이 x 폭 x 높이 / 2의 식에 의해 계산하였다.After cancer cell transplantation, from when the average tumor size reached 57.0 mm 3 to the 28th day, the tumor size for each animal was measured 11 times in 3 directions using a vernier caliper, and then length x width x height / 2 It was calculated by the formula of
최종일(day 28) 결과를 보면, 대조군의 종양 성장 억제를 0%로 하였을 때, 실시예 4, 8 및 15의 화합물(25 mg/kg)의 투여군에서 각각 56.71%, 38.35% 및 40.13% (p<0.001)의 종양 성장 억제가 관찰되었다(도 2a 및 2b 참조).Looking at the results on the last day (day 28), when the tumor growth inhibition of the control group was 0%, in the groups administered with the compounds of Examples 4, 8, and 15 (25 mg/kg), 56.71%, 38.35%, and 40.13% (p) <0.001) of tumor growth inhibition was observed (see FIGS. 2A and 2B ).
4.4 일반증상 및 체중 변화 확인4.4 Confirmation of general symptoms and weight change
HeLa CCL2 암세포 이식 누드 마우스에 실시예 화합물을 반복 복강 투여시 독성 정도를 시험하기 위해, 투여기간 동안 동물 개체의 일반증상 및 체중 변화를 관찰하였다.In order to test the degree of toxicity upon repeated intraperitoneal administration of the Example compound to HeLa CCL2 cancer cell transplantation nude mice, general symptoms and body weight changes of the animals were observed during the administration period.
그 결과 시험기간 동안 용매 대조군과 비교하여 모든 약물투여군에서 통계적으로 유의미한 체중 감소는 관찰되지 않았으며 특이한 일반증상도 없었다(도 2c 참조).As a result, no statistically significant weight loss was observed in all drug administration groups compared to the solvent control group during the test period, and there were no specific general symptoms (see FIG. 2c).
4.5. 종양무게 변화 확인4.5. Change in tumor weight
약물투여 28일째에, 마지막 투여 2시간 후 마우스 안정맥(ophthalmic veins)에서 채혈하고 CO2 가스를 이용하여 마우스를 치사시켰다. 마우스 사진을 촬영한 후 종양을 분리하여 전자저울에서 무게를 측정하였다. 사진 촬영 후, 각 종양을 절반으로 나누고, 액체질소 및 포르말린에 각각 고정하였다. 약물투여 16일째에, HeLa CCL2 종양을 절제하고 무게를 측정하였다.On the 28th day of drug administration, 2 hours after the last administration, blood was collected from the mouse ophthalmic veins and the mice were killed using CO 2 gas. After taking a picture of the mouse, the tumor was isolated and weighed on an electronic scale. After photography, each tumor was divided in half and fixed in liquid nitrogen and formalin, respectively. On day 16 of drug administration, the HeLa CCL2 tumor was excised and weighed.
용매 대조군과 비교하여, 실시예 4, 8 및 15의 화합물의 투여군에서 각각 57.1%, 40.5%, 및 26.2% (p <0.001)의 종양 무게 감소가 관찰되었다(도 2d 및 2e 참조).Compared with the solvent control group, tumor weight reductions of 57.1%, 40.5%, and 26.2% (p <0.001) were observed in the groups administered with the compounds of Examples 4, 8 and 15, respectively (see FIGS. 2D and 2E ).
실험예 5: 혈장 내 안정성 시험Experimental Example 5: Stability test in plasma
본 발명에 따른 실시예 화합물의 안정성(in vivo)를 시험하기 위하여, 실시예 2, 3, 4, 8 및 15의 화합물을 정맥 투여하고 시간 경과에 따른 혈장 농도를 관찰하였다. 또한 프로카인(Procaine)이 양성 대조군으로 사용되었다. 혈장 내 안정성은 37℃에서 인간, 랫트 또는 마우스 혈장(90 μL)에 시험 화합물을 5 mM 농도로 투여하여 3회 측정되었다. 그 결과를 하기 표 8 및 도 3a 내지 3e에 나타내었다.In order to test the stability (in vivo) of the compound of Examples according to the present invention, the compounds of Examples 2, 3, 4, 8 and 15 were administered intravenously, and plasma concentrations over time were observed. Also, Procaine was used as a positive control. Plasma stability was measured three times by administering the test compound at a concentration of 5 mM to human, rat or mouse plasma (90 μL) at 37°C. The results are shown in Table 8 and FIGS. 3A to 3E below.
화합물compound 혈장plasma 잔여 % (180분 경과)% Remaining (180 Minutes Elapsed) t1/2 t 1/2
평균average 표준편차Standard Deviation (분)(minute)
실시예 2Example 2 인간human 90.4390.43 3.233.23 >180>180
랫트rat 75.8275.82 1.081.08 >180>180
마우스mouse 55.8255.82 2.072.07 >180>180
실시예 3Example 3 인간human 56.5256.52 5.225.22 >180>180
랫트rat 0.210.21 0.020.02 4.64.6
마우스mouse 0.120.12 0.010.01 5.65.6
실시예 4Example 4 인간human 49.4149.41 0.390.39 >180>180
랫트rat 67.1167.11 1.261.26 >180>180
마우스mouse 60.4460.44 3.433.43 >180>180
실시예 8Example 8 인간human 26.0526.05 0.810.81 >180>180
랫트rat 49.2549.25 0.520.52 >180>180
마우스mouse 74.1674.16 2.252.25 >180>180
실시예 15Example 15 인간human 4.204.20 1.691.69 159.9159.9
랫트rat 41.7441.74 3.503.50 133.6133.6
마우스mouse 57.2557.25 1.801.80 >180>180
프로카인procaine 인간human 0.450.45 0.030.03
랫트rat 1.891.89 0.280.28
마우스mouse 1.061.06 0.050.05
상기 표 8 및 도 3a 내지 3e에서 보듯이, 실시예 화합물들의 혈장 내 안정성이 우수하였다. As shown in Table 8 and FIGS. 3A to 3E, the stability in plasma of the Example compounds was excellent.
실험예 6: 간 미세소관 내 대사안정성 시험Experimental Example 6: Metabolic stability test in liver microtubules
인간, 랫트 및 마우스의 간 미세소관(0.5 mg protein/mL)에 NADPH(1 mM)의 존재 또는 부재 하에 시험 화합물을 1 mM 농도로 투여하고, 37℃에서 30분간 대사안정성을 시험하였다. 또한 부스피론(Buspirone)이 양성 대조군으로 사용되었다. 그 결과를 하기 표 9 및 10; 및 도 4a 내지 4e에 나타내었다.The test compound was administered at a concentration of 1 mM in the presence or absence of NADPH (1 mM) to liver microtubules (0.5 mg protein/mL) of humans, rats and mice, and metabolic stability was tested at 37° C. for 30 minutes. Also, Buspirone was used as a positive control. The results are shown in Tables 9 and 10 below; and FIGS. 4A to 4E .
간 미세소관liver microtubules 보조인자cofactor 30분 배양 후 잔여 %% remaining after 30 min incubation
실시예 2Example 2 실시예 3Example 3 실시예 4Example 4
인간human +NADPH+NADPH 8.69 ± 0.448.69 ± 0.44 12.34 ± 0.6912.34 ± 0.69 15.76 ± 0.8915.76 ± 0.89
-NADPH-NADPH 36.40 ± 1.9036.40 ± 1.90 22.26 ± 0.7222.26 ± 0.72 77.60 ± 1.3977.60 ± 1.39
랫트rat +NADPH+NADPH 2.99 ± 0.642.99 ± 0.64 1.39 ± 0.121.39 ± 0.12 3.23 ± 0.553.23 ± 0.55
-NADPH-NADPH 41.45 ± 0.9341.45 ± 0.93 8.53 ± 0.468.53 ± 0.46 69.63 ± 1.9469.63 ± 1.94
마우스mouse +NADPH+NADPH 2.84 ± 0.342.84 ± 0.34 2.50 ± 0.112.50 ± 0.11 5.48 ± 0.045.48 ± 0.04
-NADPH-NADPH 11.59 ± 1.0611.59 ± 1.06 6.81 ± 0.176.81 ± 0.17 47.95 ± 2.9347.95 ± 2.93
버퍼buffer 77.94 ± 7.0477.94 ± 7.04 89.02 ± 4.3989.02 ± 4.39 100.93 ± 11.79100.93 ± 11.79
간 미세소관liver microtubules 보조인자cofactor 30분 배양 후 잔여 %% remaining after 30 min incubation
실시예 8Example 8 실시예 15Example 15 부스피론buspirone
인간human +NADPH+NADPH 24.11 ± 0.4624.11 ± 0.46 7.94 ± 0.747.94 ± 0.74 2.21 ± 0.032.21 ± 0.03
-NADPH-NADPH 99.18 ± 1.9299.18 ± 1.92 88.63 ± 1.6188.63 ± 1.61 91.06 ± 0.4091.06 ± 0.40
랫트rat +NADPH+NADPH 4.55 ± 0.944.55 ± 0.94 3.47 ± 0.253.47 ± 0.25 0.41 ± 0.020.41 ± 0.02
-NADPH-NADPH 85.58 ± 3.3485.58 ± 3.34 94.95 ± 1.9394.95 ± 1.93 93.08 ± 1.2193.08 ± 1.21
마우스mouse +NADPH+NADPH 11.41 ± 2.4711.41 ± 2.47 4.82 ± 0.524.82 ± 0.52 0.32 ± 0.050.32 ± 0.05
-NADPH-NADPH 89.93 ± 6.7289.93 ± 6.72 86.31 ± 1.7286.31 ± 1.72 94.74 ± 0.7494.74 ± 0.74
버퍼buffer 98.98 ± 2.0798.98 ± 2.07 91.02 ± 2.8991.02 ± 2.89 --
상기 표 9 및 10; 및 도 4a 내지 4e에서 보듯이, 실시예 4 내지 6의 화합물의 대사안정성이 가장 우수하였다. Tables 9 and 10 above; And as shown in FIGS. 4a to 4e, the metabolic stability of the compounds of Examples 4 to 6 was the best.
실험예 7: 용해도 시험Experimental Example 7: Solubility test
본 발명에 따른 실시예 화합물의 용해도를 아래와 같이 시험하였다.The solubility of the example compounds according to the present invention was tested as follows.
시험물질을 정확히 칭량하여 유리 바이알에 넣은 후 목적하는 최종 용액 부피의 20%에 해당하는 용매 A를 가하고 볼텍스믹서(Vortexer) 및 초음파로 처리(sonication)하여 완전히 용해시킨 후 최종 용액 부피의 10% 또는 20%에 해당하는 용매 B를 가하고 잘 섞어주었다. 최종 용액 부피의 80%에 해당하는 용매 C를 상기 혼합용액에 적가하여 혼합하였다. 마지막으로 30초간 초음파로 처리하였다. 상기 혼합 용액은 실험에 투여직전에 제조하여 사용하였다. After accurately weighing the test substance and putting it in a glass vial, add solvent A corresponding to 20% of the desired final solution volume and completely dissolve it by using a vortexer and sonication, and then 10% of the final solution volume or Solvent B corresponding to 20% was added and mixed well. Solvent C corresponding to 80% of the final solution volume was added dropwise to the mixed solution and mixed. Finally, it was treated with ultrasound for 30 seconds. The mixed solution was prepared and used immediately before administration in the experiment.
용매 A: 디메틸아세트아미드, SigmaAldrichSolvent A: Dimethylacetamide, SigmaAldrich
용매 B: 크레모포어(Cremophor) EL, SigmaAldrichSolvent B: Cremophor EL, SigmaAldrich
용매 C: 탈이온수 중 20% HPbCD((2-히드록시프로필)-β-시클로덱스트린, SigmaAldrich)Solvent C: 20% HPbCD ((2-hydroxypropyl)-β-cyclodextrin, SigmaAldrich) in deionized water
용해도 시험 조건 및 용해도 평가 결과를 하기 표 11에 정리하였다.The solubility test conditions and solubility evaluation results are summarized in Table 11 below.
시험
화합물exam
compound 		동물
종류animal
Kinds 		투여경로
route of administration
 		목표
농도
(mg/kg)target
density
(mg/kg) 		투약
부피
(mL/kg)dosage
volume
(mL/kg) 		투약
농도
(mg/ml)dosage
density
(mg/ml) 		시료
양
(mg)sample
sheep
(mg) 		용매
(mL)menstruum
(mL) 		투여 용매
(v/v%)dosing solvent
(v/v%) 		용해도
평가
결과Solubility
evaluation
result
실시예 4Example 4 마우스mouse 복강 abdominal cavity 2020 1010 2.02.0 2.002.00 1.0001.000 용매 ASolvent A 20%20% 용해성
있음solubility
has exist
용매 BSolvent B 10%10%
용매 Csolvent C 70%70%
실시예 8Example 8 마우스mouse 복강 abdominal cavity 2020 1010 2.02.0 2.002.00 1.0001.000 용매 ASolvent A 20%20% 용해성
있음solubility
has exist
용매 BSolvent B 20%20%
용매 Csolvent C 60%60%
실시예 15Example 15 마우스mouse 복강 abdominal cavity 2020 1010 2.02.0 2.002.00 1.0001.000 용매 ASolvent A 20%20% 용해성
있음solubility
has exist
용매 BSolvent B 20%20%
용매 Csolvent C 60%60%
실험예 8: 약물동태(PK) 시험Experimental Example 8: Pharmacokinetic (PK) test
본 발명에 따른 실시예 화합물의 약물동태(PK) 파라미터를 수컷 ICR 마우스를 이용하여 아래와 같이 시험하였다(n=3).The pharmacokinetic (PK) parameters of the example compounds according to the present invention were tested using male ICR mice as follows (n=3).
약물동태(PK) 파라미터는 KineticaTM 4.4.1(Thermo Fisher Scientific사, 미국)를 이용한 혈장농도-시간 곡선의 비구획 분석에 의해 측정되었다. 그 결과를 하기 표 12-14 및 도 5a 내지 5c에 나타내었다. 하기 표에서, "F" 파라미터는 AUClast를 이용하여 계산되었고, "N/A"는 해당 없음을 의미하며, "NC"는 계산값 없음을 의미한다.Pharmacokinetic (PK) parameters were determined by noncompartmental analysis of plasma concentration-time curves using Kinetica TM 4.4.1 (Thermo Fisher Scientific, USA). The results are shown in Table 12-14 and FIGS. 5A to 5C below. In the table below, "F" parameter was calculated using AUC last , "N/A" means not applicable, and "NC" means no calculated value.
화합물compound 실시예 4Example 4
파라미터parameter 정맥, 5 mg/kgIntravenous, 5 mg/kg 경구, 20 mg/kgOral, 20 mg/kg
평균average 표준편차Standard Deviation 평균average 표준편차Standard Deviation
tmax t max hrhr N/AN/A 0.250.25 0.000.00
Cmax C max ng/mLng/mL N/AN/A 431.00431.00 19.2919.29
AUClast AUC last ng*hr/mLng*hr/mL 1479.171479.17 120.24120.24 284.79284.79 44.1044.10
AUCinf AUC inf ng*hr/mLng*hr/mL 1502.351502.35 112.33112.33 295.27295.27 51.851.8
CLCL L/hr/kgL/hr/kg 3.343.34 0.250.25 69.1469.14 12.112.1
Vss V ss L/kgL/kg 0.980.98 0.240.24 N/AN/A
Vz V z L/kgL/kg 1.881.88 0.580.58 N/AN/A
t1/2 t 1/2 hrhr 0.390.39 0.100.10 0.40.4 0.10.1
MRTinf MRT inf hrhr 0.290.29 0.070.07 0.70.7 0.10.1
FF %% N/AN/A 4.84.8 0.750.75
화합물compound 실시예 8Example 8
파라미터parameter 정맥, 5 mg/kgIntravenous, 5 mg/kg 경구, 20 mg/kgOral, 20 mg/kg
평균average 표준편차Standard Deviation 평균average 표준편차Standard Deviation
tmax t max hrhr N/AN/A 0.580.58 0.140.14
Cmax C max ng/mLng/mL N/AN/A 93.8793.87 5.195.19
AUClast AUC last ng*hr/mLng*hr/mL 2377.942377.94 112.06112.06 96.2796.27 9.589.58
AUCinf AUC inf ng*hr/mLng*hr/mL 2386.742386.74 112.22112.22 102.54102.54 8.28.2
CLCL L/hr/kgL/hr/kg 2.102.10 0.100.10 195.92195.92 16.116.1
Vss V ss L/kgL/kg 0.500.50 0.030.03 N/AN/A
Vz V z L/kgL/kg 0.850.85 0.050.05 N/AN/A
t1/2 t 1/2 hrhr 0.280.28 0.010.01 0.40.4 0.00.0
MRTinf MRT inf hrhr 0.240.24 0.000.00 0.90.9 0.10.1
FF %% N/AN/A 1.01.0 0.100.10
화합물compound 실시예 15Example 15
파라미터parameter IV, 5 mg/kgIV, 5 mg/kg PO, 20 mg/kgPO, 20 mg/kg
평균average 표준편차Standard Deviation 평균average 표준편차Standard Deviation
tmax t max hrhr N/AN/A 0.250.25 0.000.00
Cmax C max ng/mLng/mL N/AN/A 34.4334.43 27.3427.34
AUClast AUC last ng*hr/mLng*hr/mL 1717.551717.55 41.3441.34 18.0018.00 18.4018.40
AUCinf AUC inf ng*hr/mLng*hr/mL 1729.141729.14 41.2441.24 NCNC NCNC
CLCL L/hr/kgL/hr/kg 2.892.89 0.070.07 NCNC NCNC
Vss V ss L/kgL/kg 0.710.71 0.020.02 N/AN/A
Vz V z L/kgL/kg 1.371.37 0.090.09 N/AN/A
t1/2 t 1/2 hrhr 0.330.33 0.020.02 NCNC NCNC
MRTinf MRT inf hrhr 0.240.24 0.010.01 NCNC NCNC
FF %% N/AN/A 0.30.3 0.270.27
상기 표 12-14 및 도 5a 내지 5c에서 보듯이, 실시예 화합물들은 약물로서 사용 가능한 안정성을 가짐을 확인할 수 있었다. As shown in Table 12-14 and FIGS. 5A to 5C, it was confirmed that the Example compounds had stability that could be used as drugs.
실험예 9: 암 전이에 대한 억제 효과 확인Experimental Example 9: Confirmation of inhibitory effect on cancer metastasis
a. 상처회복시험(wound healing assay)a. wound healing assay
HeLa CCL2를 6 웰에 100%로 배양하고, 옐로우팁(yellow tip)으로 상처를 가한 후, 실시예 4의 화합물을 50 nM, 100 nM 처리하였다. 약물의 효과를 관찰하기 위하여 처리전 세포의 상태를 사진으로 기록하고, 약물 처리 후 24, 36, 48 시간에 세포의 상태와 비교하여, 세포의 이동 정도를 비교 관찰하였다. 세포에 처리한 실시예 4의 화합물의 농도는 MTT를 수행하여 정하였다.HeLa CCL2 was cultured in 6 wells at 100%, and after applying a wound with a yellow tip, the compound of Example 4 was treated with 50 nM and 100 nM. In order to observe the effect of the drug, the state of the cells before treatment was recorded as a photograph, and compared with the state of the cells at 24, 36, and 48 hours after the drug treatment, the degree of cell migration was comparatively observed. The concentration of the compound of Example 4 treated with the cells was determined by performing MTT.
그 결과 도 6a 및 6b에서 보듯이, 실시예 4의 화합물의 농도가 100 nM까지는 세포의 성장에 별다른 영향을 주지 않았다.As a result, as shown in Figures 6a and 6b, the concentration of the compound of Example 4 did not have a significant effect on cell growth up to 100 nM.
b. 침투시험(in vitro invasion assay)b. In vitro invasion assay
또한, 세포의 이동을 저해하는 효능을 다시 확인하기 위해, 침투 시험(invasion assay)을 수행하였다. 24 웰 플레이트용 트랜스 웰 챔버(trans well chamber)를 준비하고, 상부 챔버(upper chamber)에 5 X 104 의 HeLa CCL2를 넣고 실시예 4의 화합물 10 nM, 50 nM, 100 nM, 250 nM 처리하고 배양한 후, 16시간 이후 세포 염색을 통해, 이동한 세포를 확인하고 그 수를 측정하였다.In addition, in order to confirm the efficacy of inhibiting cell migration again, an invasion assay was performed. Prepare a trans well chamber for a 24-well plate , put 5 X 10 4 of HeLa CCL2 in the upper chamber, and process the compound of Example 4 10 nM, 50 nM, 100 nM, 250 nM After incubation, after 16 hours, the migrated cells were identified and the number was measured through cell staining.
그 결과 도 7a 및 7b에서 보듯이, 본 발명에 따른 실시예 4의 화합물은 암 전이 억제 효과가 우수함을 확인할 수 있었다. As a result, as shown in FIGS. 7a and 7b, it was confirmed that the compound of Example 4 according to the present invention had an excellent cancer metastasis inhibitory effect.
실험예 10: 시간에 따른 각 장기별 타겟팅 측정Experimental Example 10: Targeting measurement for each organ according to time
실시예 4의 화합물에 대해 아래 절차에 따라 시간에 따른 각 장기별 타겟팅을 측정하였다.For the compound of Example 4, targeting was measured for each organ over time according to the procedure below.
각 조직의 무게를 재고 무게의 3배 양의 증류수를 넣어주었다. 호모게나이저를 이용하여 조직을 완전히 분쇄한 후 라이브러리 튜브에 조직분쇄물 50 μL와 아세토나이트릴 150 μL를 넣어주었다. 1분 동안 볼택싱(vortexing)한 후 3000 rpm, 4℃에서 10분간 원심분리하였다. 상층액을 96 웰 플레이트에 150 μL씩 넣어준 후 LC-MS/MS로 분석하였다(LC; Agilent 1260 series, MS; 3200 QTRAP, Mass polarity; positive mode, Column; Waters Xterra MS C18 2.1 X 50mm, 5 μM). 이동상은 0.1% 포름산이 들어있는 증류수(A)와 아세토나이트릴(B)을 사용하였다. LC의 변화도는 0분에 A 95%, B 5%로 시작하여, 1분에 A 5%, B 95%, 5.5분에 A 95%, B 5%의 비율 조건으로 10분 동안 실행하였다. 분석 후 LC 3.8분에서 분석값을 얻었고 MS를 이용하여 확인하였다. 그 결과를 하기 표 15 및 도 8에 나타내었다.Each tissue was weighed and distilled water in an amount three times the weight was added. After completely pulverizing the tissue using a homogenizer, 50 µL of the tissue mass and 150 µL of acetonitrile were added to the library tube. After vortexing for 1 minute, centrifugation was performed at 3000 rpm and 4° C. for 10 minutes. 150 μL of the supernatant was added to a 96-well plate and analyzed by LC-MS/MS (LC; Agilent 1260 series, MS; 3200 QTRAP, Mass polarity; positive mode, Column; Waters Xterra MS C18 2.1 X 50mm, 5 μM). As the mobile phase, distilled water (A) and acetonitrile (B) containing 0.1% formic acid were used. The gradient of LC started with A 95% and B 5% at 0 min, and was carried out for 10 min under the condition of A 5%, B 95% at 1 min, A 95%, B 5% at 5.5 min. After analysis, an analysis value was obtained at LC 3.8 min and confirmed using MS. The results are shown in Table 15 and FIG. 8 below.
구분division 시간(hr)time (hr) 실시예 4의 화합물의 농도 (ng/mL)Concentration of the compound of Example 4 (ng/mL)
#1#One #2#2 #3#3 MeanMean SDSD
암조직
(Tumor)cancer tissue
(Tumor) 		33 33.033.0 39.939.9 44.044.0 39.0 39.0 5.5 5.5
66 17.617.6 18.718.7 29.429.4 21.9 21.9 6.6 6.6
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
간
(Liver)liver
(Liver) 		33 11.411.4 12.412.4 12.512.5 12.1 12.1 0.6 0.6
66 9.99.9 9.79.7 10.510.5 10.0 10.0 0.4 0.4
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
지라
(Spleen)jira
(Spleen) 		33 10.810.8 9.89.8 11.011.0 10.5 10.5 0.7 0.7
66 <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
혈장
(Plasma)plasma
(Plasma) 		33 <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
66 <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
심장
(Heart)heart
(Heart) 		33 <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ
66 <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ
신장
(Kidney)kidney
(Kidney) 		33 14.714.7 11.411.4 37.037.0 21.0 21.0 14.0 14.0
66 9.79.7 11.211.2 11.211.2 10.7 10.7 0.9 0.9
미처리unprocessed <LLOQ<LLOQ <LLOQ<LLOQ <LLOQ<LLOQ 　 　
* 최소검출한계(LLOQ) 3.9 ng/mL상기 표 15 및 도 8에서 보듯이, 실시예 4의 화합물은 여러 장기 중에서 암 조직을 효과적으로 타겟팅하는 것을 확인할 수 있었다.* Minimum limit of detection (LLOQ) 3.9 ng/mL As shown in Table 15 and FIG. 8, it was confirmed that the compound of Example 4 effectively targeted cancer tissues among various organs.
실험예 11: 약물-형광체 접합체의 타겟팅 평가Experimental Example 11: Targeting evaluation of drug-phosphor conjugates
약물-형광체 접합체로서 실시예 30의 N-아실히드라존 유도체-형광체 접합체의 암조직에 대한 타겟팅을 평가하였다.Targeting of the N-acylhydrazone derivative-phosphor conjugate of Example 30 to cancer tissue was evaluated as a drug-phosphor conjugate.
먼저 형광체(Cy5)만의 세포내 투과를 시험하고 실시예 30의 접합체의 세포내 투과를 시험하여 그 결과를 도 9에 나타내었다. 또한 4',6-디아미디노-2-페닐인돌(DAPI)을 이용한 시험 결과도 함께 나타내었고, 이로부터 상기 접합체의 세포내 투과를 확인할 수 있다. First, intracellular permeation of only the phosphor (Cy5) was tested, and intracellular permeation of the conjugate of Example 30 was tested, and the results are shown in FIG. 9 . In addition, the test results using 4',6-diamidino-2-phenylindole (DAPI) were also shown, from which intracellular permeation of the conjugate can be confirmed.
다음으로 고형 종양 이식(xenograft) 마우스 모델을 만들기 위해 누드 마우스의 상박 어깨 부위에 5x106의 HeLa CCL2를 접종하였다. 종양의 크기가 100~120 mm3가 된 후 실시예 30의 N-아실히드라존 유도체-형광체 접합체 5 mg/kg를 꼬리에 정맥 주사하였다. 형광 영상 장비로서 IVIS(in vivo Imaging system spectrum, PerkinElmer)를 이용하여 640 nm 여기 및 700 nm 배출 필터(680, 700, 720 nm for spectral unmixing, binnig factor = 8) 조건에서 6, 24, 30 및 48 시간에 촬영하였다. 촬영이 끝난 후 마우스 모델을 희생하여 장기를 적출하였고, 24 및 48 시간에 생체 외 촬영을 진행하였다. Next, in order to make a solid tumor transplantation (xenograft) mouse model, 5x10 6 HeLa CCL2 was inoculated into the upper arm of nude mice. After the tumor became 100-120 mm 3 in size, 5 mg/kg of the N-acylhydrazone derivative-fluorescent conjugate of Example 30 was intravenously injected into the tail. 6, 24, 30 and 48 under 640 nm excitation and 700 nm emission filters (680, 700, 720 nm for spectral unmixing, binnig factor = 8) using IVIS (in vivo Imaging system spectrum, PerkinElmer) as a fluorescence imaging device taken on time. After imaging was completed, the mouse model was sacrificed and organs were removed, and ex vivo imaging was performed at 24 and 48 hours.
도 10에서 보듯이, 생체 내 촬영 결과 시간이 경과함에 따라 종양의 형광 세기가 점점 더 강하게 보이는 것이 관찰되는 한편, 일반 배경과 다른 장기들의 형광 세기는 감소되는 것이 관찰되었다. 도 11에서 보듯이, 각 조직별(심장, 폐, 간, 지라, 신장, 암조직) 생체 외 촬영에서도 24 및 48 시간 모두에서 종양의 높은 형광을 확인할 수 있었다. 구체적으로 물질 배설을 담당하고 있는 신장을 제외하고 그 다음으로 종양에서 가장 높은 형광 세기를 보였고, 도 12의 정량값 비교에서도 비슷한 상관관계를 보였다. 또한 도 13에서 보듯이, 종양과 일반 장기인 마우스 대퇴부 부위 근육층의 형광 세기를 정량 비교한 결과 유의한 차이를 확인할 수 있었다. 이를 통해 실시예 30의 접합체가 종양을 정확히 타겟팅하는 것을 확인할 수 있었다. As shown in FIG. 10 , as a result of in vivo imaging, it was observed that the fluorescence intensity of the tumor gradually increased over time, while the fluorescence intensity of organs other than the general background decreased. As shown in FIG. 11 , high fluorescence of the tumor was confirmed at both 24 and 48 hours in the in vitro imaging of each tissue (heart, lung, liver, spleen, kidney, cancer tissue). Specifically, except for the kidney responsible for excretion of substances, the tumor showed the highest fluorescence intensity, and a similar correlation was also observed in the comparison of quantitative values in FIG. 12 . Also, as shown in FIG. 13 , a significant difference was confirmed as a result of quantitative comparison of the fluorescence intensity of the muscle layer of the thigh region of the mouse, which is a general organ, with the tumor. Through this, it was confirmed that the conjugate of Example 30 accurately targeted the tumor.
실험예 12: 약물-리간드 접합체의 타겟팅 평가Experimental Example 12: Targeting evaluation of drug-ligand conjugates
약물-리간드 접합체로서 실시예 29의 N-아실히드라존 유도체-리간드 접합체의 암조직에 대한 타겟팅을 평가하였다.As a drug-ligand conjugate, the targeting of the N-acylhydrazone derivative-ligand conjugate of Example 29 to cancer tissues was evaluated.
1. 89Zr 결합된 시약의 제조1. Preparation of 89 Zr-bound reagent
약물-리간드 접합체 1 mg을 약간의 DMSO에 녹인 후 인산완충식염수(PBS)로 희석하였다. 이때 접합체가 석출되면 DMSO 양을 늘리고, 최종 부피를 1 mL에 맞추었다. 수득한 1 mL 용액에 89Zr4+ 300 μCi를 넣고 1시간 이상 500 rpm으로 교반하였다. 이후 PBS로 활성화시킨 PD 10 컬럼으로 옮기고 PBS를 1 mL씩 흘려주며 용액을 받았다. 용액을 최대 10 mL을 흘려주며 1 mL씩 튜브로 받아 방사성 값을 측정하였다. 4~5번째 튜브에 나온 용액이 가장 순도가 좋았고, 뒤로 갈수록 튜브에는 접합체와 결합하지 못한 89Zr4+가 많았다. 가장 높은 방사성 값이 나온 튜브를 이용하여 이종이식 마우스 모델(Xenograft mouse model)의 꼬리에 최대 200 μCi 및 최소 50 μCi로 정맥주사하였다(매번 접합마다 방사성의 값이 달라지므로 주입량이 정해지면 모든 마우스를 동일 방사성 양으로 주사하였다). 1 mg of the drug-ligand conjugate was dissolved in a little DMSO and then diluted with phosphate buffered saline (PBS). At this time, when the conjugate was precipitated, the amount of DMSO was increased, and the final volume was adjusted to 1 mL. 89 Zr 4+ 300 μCi was added to the obtained 1 mL solution and stirred at 500 rpm for at least 1 hour. After that, it was transferred to a PD 10 column activated with PBS, and 1 mL of PBS was flowed to receive the solution. A maximum of 10 mL of the solution was flowed, and the radioactivity value was measured by receiving 1 mL each into a tube. The solution from the 4th to 5th tube had the highest purity, and as it went further back, the tube contained more 89 Zr 4+ that could not be combined with the conjugate. Using the tube with the highest radioactivity value, it was intravenously injected into the tail of a xenograft mouse model at a maximum of 200 μCi and a minimum of 50 μCi (as the radioactivity value varies for each junction, all mice were treated when the injection amount was determined. were injected with the same radioactive dose).
2. PET 분석2. PET Analysis
앞서 제조한 시약을 종양 크기가 100~120 mm3인 이종이식(xenograft) 마우스 모델에 주입한 후 양성자방출 단층촬영(PET) 장비로 1, 24, 48, 72 및 96 시간에 스캔하였다(1800 sec, IAW, Siemens, USA). 이후 소프트웨어(IRW, Siemens, USA)를 이용해 영상 이미징 작업을 하여 ROI(Region of interest) 정량값을 얻었다. 도 14에서 보듯이, 마우스 우측 대퇴부의 고형 종양에 실시예 29의 접합체가 축적된 것이 관찰되었고 최대 96시간까지 관찰할 수 있었다. 일반 장기와 배경들의 동위원소 신호가 시간이 지남에 따라 줄어들어 종양의 축적을 더 정확히 관찰할 수 있었다. 약물에서 떨어져 나와 자유로운 상태가 된 89Zr4+는 뼈에 잘 부착이 되기 때문에 척추나 관절 등에 많이 축적된 것이 관찰되었고 외부 물질을 흡수하여 대사작용을 하는 간 또한 높은 축적을 보였다. 정량값 또한 96 시간까지 유의한 값을 나타냈다. After injecting the previously prepared reagent into a xenograft mouse model with a tumor size of 100 to 120 mm 3 , it was scanned with proton emission tomography (PET) at 1, 24, 48, 72 and 96 hours (1800 sec). , IAW, Siemens, USA). Then, image imaging was performed using software (IRW, Siemens, USA) to obtain a quantitative value of ROI (Region of Interest). As shown in FIG. 14 , it was observed that the conjugate of Example 29 was accumulated in a solid tumor of the right thigh of the mouse, and it could be observed for up to 96 hours. The isotopic signal of common organs and backgrounds decreased over time, allowing for more accurate observation of tumor accumulation. 89 Zr 4+ , which was released from the drug and became free, was observed to accumulate a lot in the spine and joints because it adheres well to the bones. Quantitative values also showed significant values up to 96 hours.
스캔이 끝난 마우스를 희생하여 장기들을 적출한 뒤 생체 외 실험을 진행하였다. PET 영상 촬영 시간과 같은 1, 24, 48, 72 및 96 시간에 마우스를 희생하였고 감마계측기(gamma counter, Wizard2, PerkinElmer)로 각각의 장기의 정량값을 측정하였다. 도 15에서 보듯이 종양에서 실시예 29의 접합체가 높은 수치로 측정되었다. 이와 같은 결과를 통해 약물-접합체 리간드가 종양을 정확히 타겟팅 한다는 것을 확인할 수 있었고 앞선 실험인 약물-형광제 접합체와도 같은 결과를 보여 약물의 종양 타켓팅 실험이 유의하다는 것을 한 번 더 확인할 수 있었다. 즉, 본원 발명에 따른 N-아실히드라존 유도체와 표지자의 접합체의 종양 타겟팅에 관한 우수한 효과를 확인하였다.After the scan was completed, the mice were sacrificed and organs were removed, followed by in vitro experiments. Mice were sacrificed at 1, 24, 48, 72 and 96 hours at the same time as PET imaging, and quantitative values of each organ were measured with a gamma counter (Wizard2, PerkinElmer). As shown in FIG. 15 , the conjugate of Example 29 was measured at a high level in the tumor. Through these results, it was confirmed that the drug-conjugate ligand accurately targets the tumor, and the same results as the previous experiment, the drug-fluorescent conjugate, were shown, confirming once more that the drug tumor targeting experiment was significant. That is, the excellent effect on tumor targeting of the conjugate of the N-acylhydrazone derivative and the marker according to the present invention was confirmed.
실험예 13: 약물-리간드 접합체와 다른 약물 간의 타겟팅 경쟁 평가Experimental Example 13: Evaluation of targeting competition between drug-ligand conjugates and other drugs
약물-리간드 접합체와 같은 메커니즘으로 종양을 억제시키는 다른 약물들과의 경쟁 유/무를 확인하였다. 약물-리간드 접합체로서 실시예 29의 N-아실히드라존-리간드 접합체(실시예 4의 화합물과 DFO의 접합체)를 사용하였다. 1x105의 HeLa CCL2를 12 웰 플레이트에서 24 시간 동안에 배양하고, 약물-리간드 접합체를 단독으로 또는 경쟁 약물과 동시에 투약한 후 1, 2 및 4 시간에 배지를 제거한 후 PBS로 암세포를 세척하였다. 1 N NaOH로 암세포를 용해한 후 투명한 테스트 튜브에 넣고 감마계측계로 정량값을 측정하였다.The presence/absence of competition with other drugs that suppress tumors by the same mechanism as drug-ligand conjugates was confirmed. As the drug-ligand conjugate, the N-acylhydrazone-ligand conjugate of Example 29 (conjugate of the compound of Example 4 with DFO) was used. 1x10 5 HeLa CCL2 was cultured in a 12-well plate for 24 hours, and the medium was removed 1, 2, and 4 hours after the drug-ligand conjugate was administered alone or simultaneously with a competing drug, and then the cancer cells were washed with PBS. After dissolving cancer cells with 1 N NaOH, it was placed in a transparent test tube and quantitative values were measured with a gamma meter.
도 16에서 보듯이, 경쟁 약물이 없는 경우(검은 막대)에는 시간이 지남에 따라 실시예 29의 접합체가 암세포 내로 흡수(uptake)되는 양이 증가하였다. 그러나 실시예 29의 접합체와 실시예 4의 화합물을 동시에 투약하는 경우(흰색 막대)에는 시간이 지남에 따라 접합체의 암세포 내 흡수가 증가하지 않았고 오히려 줄어드는 것을 관찰할 수 있었다. 또한 도 17에서 보듯이, 실시예 29의 접합체를 단독으로 투약하는 경우(DMSO)에 대비하여, 다른 약물(실시예 3, 4, 5, 18, 20, 21, 22, 23, 24, 25, 26의 화합물)과 동시에 투약하는 경우에 경쟁에 의해 세포 내 흡수가 차단/억제되는 것을 확인할 수 있었다.As shown in FIG. 16 , in the absence of a competing drug (black bars), the amount of the conjugate of Example 29 uptake into cancer cells increased over time. However, when the conjugate of Example 29 and the compound of Example 4 were administered at the same time (white bar), it was observed that the uptake of the conjugate into cancer cells did not increase over time, but rather decreased. In addition, as shown in FIG. 17, in contrast to the case of administering the conjugate of Example 29 alone (DMSO), other drugs (Examples 3, 4, 5, 18, 20, 21, 22, 23, 24, 25, 26), it was confirmed that intracellular uptake was blocked/inhibited by competition.
이와 같이 실시예 29의 접합체가 암세포를 표적하여 세포안으로 흡수되는 한편, 같은 매커니즘을 갖는 경쟁 약물에 의해 억제되는 것이 관찰되었으므로, 실시예 29의 접합체와 경쟁 약물이 암세포 안으로 들어가는 메카니즘 면에서 같음을 확인할 수 있었다. As described above, it was observed that the conjugate of Example 29 was absorbed into the cells by targeting the cancer cells while being inhibited by the competing drug having the same mechanism. could

Claims (29)

  1. 하기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:A conjugate of an N-acylhydrazone derivative of Formula 1 and a marker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
    [화학식 1][Formula 1]
    Figure PCTKR2021007223-appb-img-000123
    Figure PCTKR2021007223-appb-img-000123
    상기 식에서in the above formula
    R은
    Figure PCTKR2021007223-appb-img-000124
    ,
    Figure PCTKR2021007223-appb-img-000125
    ,
    Figure PCTKR2021007223-appb-img-000126
    또는
    Figure PCTKR2021007223-appb-img-000127
    이고;    R is
    Figure PCTKR2021007223-appb-img-000124
    ,
    Figure PCTKR2021007223-appb-img-000125
    ,
    Figure PCTKR2021007223-appb-img-000126
    or
    Figure PCTKR2021007223-appb-img-000127
    ego;
    A는 -O- 또는 -S-이고;A is -O- or -S-;
    R1은 H, C1-6알킬, C1-6알콕시카보닐C1-3알킬, 또는 C1-6알콕시C1-3알킬이고;R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl;
    R2는 할로겐, C1-6알킬 또는 할로C1-6알킬이고;R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl;
    R3은 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 할로C1-6알콕시이고;R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy;
    R4 및 R5는 각각 독립적으로 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 C1-6알킬카보닐아미노이고,R 4 and R 5 are each independently H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or C 1-6 alkylcarbonylamino;
    상기 표지자는 방사성 동위원소의 리간드 또는 형광체이다.The marker is a ligand or fluorescent substance of a radioisotope.
  2. 제1항에 있어서,The method of claim 1,
    R1은 H, -CH3, -CH2CO2CH3, -CH2CO2CH2CH3, -CH2CH2OCH2CH3 또는 -CH2CH2OCH3이고;R 1 is H, —CH 3 , —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 , —CH 2 CH 2 OCH 2 CH 3 or —CH 2 CH 2 OCH 3 ;
    R2는 Cl, Br, -CH3 또는 -CF3이고;R 2 is Cl, Br, —CH 3 or —CF 3 ;
    R3은 H, F, Cl, -CH3, -OCH3, 또는 -OCF3이고;R 3 is H, F, Cl, -CH 3 , -OCH 3, or -OCF 3, and;
    R4 및 R5는 각각 독립적으로 H, Cl, -CH3, -OCH3, 또는 -NHCOCH3인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.R 4 and R 5 are each independently H, Cl, -CH 3, -OCH 3, -NHCOCH 3 or a conjugate thereof, stereoisomers or a pharmaceutically acceptable salt thereof.
  3. 제1항에 있어서,According to claim 1,
    R1은 -CH2CO2CH3, -CH2CO2CH2CH3 또는 -CH2CH2OCH2CH3이고;R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ;
    R2는 Cl, Br 또는 -CH3이고;R 2 is Cl, Br or —CH 3 ;
    R3은 H 또는 -OCH3이고;R 3 is H or —OCH 3 ;
    R4 및 R5는 각각 독립적으로 H, Cl, -CH3 또는 -OCH3인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.R 4 and R 5 are each independently H, Cl, -CH 3 or -OCH 3 A conjugate, a stereoisomer, or a pharmaceutically acceptable salt thereof.
  4. 제1항에 있어서,According to claim 1,
    상기 화학식 1의 N-아실히드라존 유도체가 하기 중에서 선택되는 어느 하나인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:A conjugate in which the N-acylhydrazone derivative of Formula 1 is any one selected from the following, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
    (E)-N'-[(2-클로로-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-[(2-클로로-1-메틸-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
    에틸 (E)-2-{2-클로로-3-[(2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸]-1H-인돌-1-일}아세테이트;ethyl (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl]-1H-indol-1-yl}acetate;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-브로모-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-메톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydra Gide;
    (E)-5-클로로-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}벤조퓨란-2-카보히드라지드;(E)-5-chloro-N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,7-디메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,6-디메톡시벤조퓨란-2-카보히드라지드; (E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbohydrazide;
    (E)-N'-{2-[2-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌]히드라진-1-카보닐}벤조퓨란-5-일)아세트아미드;(E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl}benzofuran-5 -yl)acetamide;
    (E)-에틸-2-(3-((2-(4,6-디메톡시벤조퓨란-2-카보닐)히드라진일리덴)메틸)-2-메틸-1H-인돌-1-일)아세테이트;(E)-ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazinylidene)methyl)-2-methyl-1H-indol-1-yl)acetate ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbohydrazide;
    (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트; 및(E)-ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl) acetate; and
    (E)-메틸 2-(2-메틸-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트.(E)-methyl 2-(2-methyl-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl) acetate.
  5. 제1항에 있어서, According to claim 1,
    상기 N-아실히드라존 유도체의 R1의 말단이 아민화되어 상기 표지자와 접합되는 것인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.A conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the terminal of R 1 of the N-acylhydrazone derivative is aminated to be conjugated with the marker.
  6. 제5항에 있어서, 6. The method of claim 5,
    상기 N-아실히드라존 유도체는 링커를 매개로 상기 표지자와 접합되거나 링커 없이 직접 결합이며,The N-acylhydrazone derivative is conjugated to the marker via a linker or is a direct bond without a linker,
    상기 링커는 p-NCS-벤젠을 포함하는 접합체 또는 -C(=O)-이며, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.The linker is a conjugate comprising p-NCS-benzene or -C(=O)-, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  7. 제1항에 있어서, According to claim 1,
    상기 방사성 동위원소의 리간드는 디페록사민(DFO), 디에틸렌트리아민펜타아세트산(DTPA) 및 1,4,7,10-테트라아자시클로도데칸-1,4,7,10-테트라아세트산(DOTA)로 이루어진 군에서 선택되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.Ligands of the radioisotope include deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). ) a conjugate selected from the group consisting of, a stereoisomer or a pharmaceutically acceptable salt thereof.
  8. 제1항에 있어서, According to claim 1,
    상기 방사성 동위원소는 89Zr, 60Cu, 64Cu, 223Ra, 131I, 89Sr, 32P 및 188Re로 이루어진 군에서 선택되는 1종 이상인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.The radioactive isotope is at least one conjugate selected from the group consisting of 89 Zr, 60 Cu, 64 Cu, 223 Ra, 131 I, 89 Sr, 32 P and 188 Re, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. .
  9. 제1항에 있어서, According to claim 1,
    상기 형광체는 시아닌 3, 시아닌 5, 시아닌 5.5 및 시아닌 7로 이루어진 군에서 선택되는 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.The phosphor is a conjugate selected from the group consisting of cyanine 3, cyanine 5, cyanine 5.5 and cyanine 7, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  10. 제1항에 있어서,According to claim 1,
    상기 화학식 1의 N-아실히드라존 유도체와 표지자의 접합체는 The conjugate of the N-acylhydrazone derivative of Formula 1 and the marker is
    하기 화학식 2로 표시되는 접합체인, 접합체 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:A conjugate represented by the following formula (2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
    [화학식 2][Formula 2]
    Figure PCTKR2021007223-appb-img-000128
    Figure PCTKR2021007223-appb-img-000128
    상기 식에서 in the above formula
    R, R2, R3은 상기 화학식 1에서 정의된 바와 같고;R, R 2 , R 3 are as defined in Formula 1 above;
    R0은 상기 화학식 1의 R1에서 하나의 H가 제거된 그룹이고;R 0 is a group in which one H is removed from R 1 in Formula 1;
    L은 단일 결합, 또는 링커로부터 유래된 그룹이고;L is a single bond or a group derived from a linker;
    A는 방사성 동위원소의 리간드 또는 형광체로부터 유래된 그룹이다.A is a group derived from a ligand or fluorescent substance of a radioisotope.
  11. 제10항에 있어서,11. The method of claim 10,
    R0는 -(CH2)m-, -(CH2)n-COO-(CH2)m-, 또는 -(CH2)n-O-(CH2)m-이고;R 0 is -(CH 2 ) m -, -(CH 2 ) n -COO-(CH 2 ) m -, or -(CH 2 ) n -O-(CH 2 ) m -;
    m은 1 내지 6의 정수이고, n은 1 내지 3의 정수이고;m is an integer from 1 to 6, n is an integer from 1 to 3;
    L은 단일 결합, -C(=O)- 또는 -C(=S)-NH-페닐렌-NH-C(=S)-이고;L is a single bond, -C(=O)- or -C(=S)-NH-phenylene-NH-C(=S)-;
    A는 하기로 이루어진 군으로부터 선택되는 어느 하나인, 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:A is any one selected from the group consisting of a conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof:
    Figure PCTKR2021007223-appb-img-000129
    ,
    Figure PCTKR2021007223-appb-img-000129
    ,
    Figure PCTKR2021007223-appb-img-000130
    ,
    Figure PCTKR2021007223-appb-img-000130
    ,
    Figure PCTKR2021007223-appb-img-000131
    ,
    Figure PCTKR2021007223-appb-img-000131
    ,
    Figure PCTKR2021007223-appb-img-000132
    ,
    Figure PCTKR2021007223-appb-img-000132
    ,
    Figure PCTKR2021007223-appb-img-000133
    ,
    Figure PCTKR2021007223-appb-img-000133
    ,
    Figure PCTKR2021007223-appb-img-000134
    및
    Figure PCTKR2021007223-appb-img-000134
    and
    ,,
    Figure PCTKR2021007223-appb-img-000135
    .
    Figure PCTKR2021007223-appb-img-000135
    .
  12. 제11항에 있어서, R0는 -CH2, -CH2CO2CH2, -CH2CO2CH2CH2, -CH2CH2OCH2CH2 또는 -CH2CH2OCH2인, 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.12. The method of claim 11 , wherein R 0 is —CH 2 , —CH 2 CO 2 CH 2 , —CH 2 CO 2 CH 2 CH 2 , —CH 2 CH 2 OCH 2 CH 2 or —CH 2 CH 2 OCH 2 , A conjugate, a stereoisomer or a pharmaceutically acceptable salt thereof.
  13. 제10항에 있어서, N-아실히드라존 유도체가 하기 중에서 선택되는 어느 하나인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:The conjugate according to claim 10, wherein the N-acylhydrazone derivative is any one selected from the following, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
    (E)-N'-[(2-클로로-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-[(2-chloro-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-[(2-클로로-1-메틸-1H-인돌-3-일)메틸렌]-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-[(2-chloro-1-methyl-1H-indol-3-yl)methylene]-5-methylbenzofuran-2-carbohydrazide;
    에틸 (E)-2-{2-클로로-3-[(2-(5-메틸벤조퓨란-2-카보닐)히드라지닐리덴)메틸]-1H-인돌-1-일}아세테이트;ethyl (E)-2-{2-chloro-3-[(2-(5-methylbenzofuran-2-carbonyl)hydrazinylidene)methyl]-1H-indol-1-yl}acetate;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-브로모-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-bromo-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[1-(2-에톡시에틸)-2-(트리플루오로메틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[1-(2-ethoxyethyl)-2-(trifluoromethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-메톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-methoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-6-메톡시-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-6-methoxy-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-플루오로-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-fluoro-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide ;
    (E)-N'-{[2,5-디클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2,5-dichloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-(트리플루오로메톡시)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-(trifluoromethoxy)-1H-indol-3-yl]methylene}-5-methylbenzofuran-2 -carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메틸-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-5-methyl-1H-indol-3-yl]methylene}-5-methylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일]메틸렌}-5-메톡시벤조퓨란-2-카보히드라지드;(E)-N'-{[2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl]methylene}-5-methoxybenzofuran-2-carbohydra Gide;
    (E)-5-클로로-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}벤조퓨란-2-카보히드라지드;(E)-5-chloro-N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,7-디메틸벤조퓨란-2-카보히드라지드;(E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,7-dimethylbenzofuran-2-carbohydrazide;
    (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-4,6-디메톡시벤조퓨란-2-카보히드라지드; (E)—N′-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-4,6-dimethoxybenzofuran-2-carbohydrazide;
    (E)-N'-{2-[2-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌]히드라진-1-카보닐}벤조퓨란-5-일)아세트아미드;(E)-N'-{2-[2-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene]hydrazine-1-carbonyl}benzofuran-5 -yl)acetamide;
    (E)-에틸-2-(3-((2-(4,6-디메톡시벤조퓨란-2-카보닐)히드라진일리덴)메틸)-2-메틸-1H-인돌-1-일)아세테이트;(E)-ethyl-2-(3-((2-(4,6-dimethoxybenzofuran-2-carbonyl)hydrazinylidene)methyl)-2-methyl-1H-indol-1-yl)acetate ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드; (E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbohydrazide;
    (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트; 및 (E)-ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl) acetate; and
    (E)-메틸 2-(2-메틸-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트. (E)-methyl 2-(2-methyl-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl) acetate.
  14. 제13항에 있어서, 상기 N-아실히드라존 유도체가 (E)-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}-5-메틸벤조퓨란-2-카보히드라지드 또는 (E)-5-클로로-N'-{[2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일]메틸렌}벤조퓨란-2-카보히드라지드인 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:14. The method of claim 13, wherein the N-acylhydrazone derivative is (E)-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}-5- Methylbenzofuran-2-carbohydrazide or (E)-5-chloro-N'-{[2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl]methylene}benzofuran- A conjugate that is 2-carbohydrazide, a stereoisomer or a pharmaceutically acceptable salt thereof:
  15. 제14항에 있어서, 접합체는 하기로 이루어진 군으로부터 선택되는 어느 하나인, 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.15. The method of claim 14, wherein the conjugate is any one selected from the group consisting of, the conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
    Figure PCTKR2021007223-appb-img-000136
    ,
    Figure PCTKR2021007223-appb-img-000136
    ,
    Figure PCTKR2021007223-appb-img-000137
    ,
    Figure PCTKR2021007223-appb-img-000137
    ,
    Figure PCTKR2021007223-appb-img-000138
    ,
    Figure PCTKR2021007223-appb-img-000138
    ,
    Figure PCTKR2021007223-appb-img-000139
    ,
    Figure PCTKR2021007223-appb-img-000139
    ,
    Figure PCTKR2021007223-appb-img-000140
    ,
    Figure PCTKR2021007223-appb-img-000140
    ,
    Figure PCTKR2021007223-appb-img-000141
    , 및
    Figure PCTKR2021007223-appb-img-000141
    , and
    Figure PCTKR2021007223-appb-img-000142
    Figure PCTKR2021007223-appb-img-000142
  16. 제15항에 있어서, 접합체는 16. The method of claim 15, wherein the conjugate is
    Figure PCTKR2021007223-appb-img-000143
    Figure PCTKR2021007223-appb-img-000143
    또는 or
    Figure PCTKR2021007223-appb-img-000144
    Figure PCTKR2021007223-appb-img-000144
    인, 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.Phosphorus, a conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  17. 제1항 내지 제16항 중 어느 한 항의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 포함하는, 세포증식성 질환의 발생, 진행 또는 전이를 진단하기 위한 약학적 조성물.A pharmaceutical composition for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising the conjugate of any one of claims 1 to 16, a stereoisomer or a pharmaceutically acceptable salt thereof.
  18. 제17항에 있어서, 18. The method of claim 17,
    상기 조성물은 조영제인, 약학적 조성물.The composition is a contrast agent, a pharmaceutical composition.
  19. 제17항에 있어서,18. The method of claim 17,
    상기 조성물은 세포증식성 질환의 치료 효과를 또한 나타내는, 약학적 조성물.The composition also exhibits a therapeutic effect on a cell proliferative disease, a pharmaceutical composition.
  20. 제17항에 있어서,18. The method of claim 17,
    상기 세포증식성 질환은 직장암, 유방암, 폐암, 위암, 간암, 백혈병, 신경교종, 피부암, 자궁경부암 및 이로부터 유래된 전이물을 포함하는, 약학적 조성물.The cell proliferative disease includes rectal cancer, breast cancer, lung cancer, gastric cancer, liver cancer, leukemia, glioma, skin cancer, cervical cancer and metastases derived therefrom, a pharmaceutical composition.
  21. 세포증식성 질환의 발생, 진행 또는 전이의 진단용 약제의 제조를 위한 제1항 내지 제16항 중 어느 한 항의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염의 용도. Use of the conjugate of any one of claims 1 to 16, a stereoisomer or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the diagnosis of the occurrence, progression or metastasis of a cell proliferative disease.
  22. 제1항 내지 제16항 중 어느 한 항의 접합체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 필요로 하는 대상에 투여하는 것을 포함하는, 세포증식성 질환의 발생, 진행 또는 전이를 진단하는 방법.A method for diagnosing the occurrence, progression or metastasis of a cell proliferative disease, comprising administering to a subject in need thereof the conjugate of any one of claims 1 to 16, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  23. 하기 화학식 1로 표시되는 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:An N-acylhydrazone derivative represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
    [화학식 1][Formula 1]
    Figure PCTKR2021007223-appb-img-000145
    Figure PCTKR2021007223-appb-img-000145
    상기 식에서in the above formula
    R은
    Figure PCTKR2021007223-appb-img-000146
    ,
    Figure PCTKR2021007223-appb-img-000147
    ,
    Figure PCTKR2021007223-appb-img-000148
    또는
    Figure PCTKR2021007223-appb-img-000149
    이고;    R is
    Figure PCTKR2021007223-appb-img-000146
    ,
    Figure PCTKR2021007223-appb-img-000147
    ,
    Figure PCTKR2021007223-appb-img-000148
    or
    Figure PCTKR2021007223-appb-img-000149
    ego;
    A는 -O- 또는 -S-이고;A is -O- or -S-;
    R1은 H, C1-6알킬, C1-6알콕시카보닐C1-3알킬, 또는 C1-6알콕시C1-3알킬이고;R 1 is H, C 1-6 alkyl, C 1-6 alkoxycarbonylC 1-3 alkyl, or C 1-6 alkoxyC 1-3 alkyl;
    R2는 할로겐, C1-6알킬 또는 할로C1-6알킬이고;R 2 is halogen , C 1-6 alkyl or haloC 1-6 alkyl;
    R3은 H, 할로겐, C1-6알킬, C1-6알콕시, 또는 할로C1-6알콕시이다.R 3 is H, halogen, C 1-6 alkyl, C 1-6 alkoxy, or haloC 1-6 alkoxy.
  24. 제23항에 있어서,24. The method of claim 23,
    R1은 -CH2CO2CH3, -CH2CO2CH2CH3 또는 -CH2CH2OCH2CH3이고;R 1 is —CH 2 CO 2 CH 3 , —CH 2 CO 2 CH 2 CH 3 or —CH 2 CH 2 OCH 2 CH 3 ;
    R2는 Cl 또는 -CH3이고;R 2 is Cl or —CH 3 ;
    R3은 H 또는 -OCH3인, N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염.R 3 is H or —OCH 3 , an N-acylhydrazone derivative, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
  25. 제23항에 있어서,24. The method of claim 23,
    하기 중에서 선택되는 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염:An N-acylhydrazone derivative selected from the following, a stereoisomer or a pharmaceutically acceptable salt thereof:
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)벤조[b]티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)benzo[b]thiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐퓨란-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylfuran-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-5-메톡시-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)-N'-((2-chloro-1-(2-ethoxyethyl)-5-methoxy-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide ;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-5-페닐티오펜-2-카보히드라지드;(E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-5-phenylthiophene-2-carbohydrazide;
    (E)-N'-((2-클로로-1-(2-에톡시에틸)-1H-인돌-3-일)메틸렌)-1-페닐-1H-피라졸-5-카보히드라지드; 및(E)—N′-((2-chloro-1-(2-ethoxyethyl)-1H-indol-3-yl)methylene)-1-phenyl-1H-pyrazole-5-carbohydrazide; and
    (E)-에틸 2-(2-클로로-3-((2-(나프토[2,1-b]퓨란-2-카보닐)히드라진일리덴)메틸)-1H-인돌-1-일)아세테이트.(E)-ethyl 2-(2-chloro-3-((2-(naphtho[2,1-b]furan-2-carbonyl)hydrazinylidene)methyl)-1H-indol-1-yl) acetate.
  26. 제23항 내지 제25항 중 어느 한 항의 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는, 세포증식성 질환의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for the prevention or treatment of cell proliferative diseases, comprising the N-acylhydrazone derivative of any one of claims 23 to 25, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  27. 제26항에 있어서,27. The method of claim 26,
    상기 세포증식성 질환은 직장암, 유방암, 폐암, 위암, 간암, 백혈병, 신경교종, 피부암, 자궁경부암 및 이로부터 유래된 전이물을 포함하는, 약학적 조성물.The cell proliferative disease includes rectal cancer, breast cancer, lung cancer, gastric cancer, liver cancer, leukemia, glioma, skin cancer, cervical cancer and metastases derived therefrom, a pharmaceutical composition.
  28. 세포증식성 질환의 예방 또는 치료에 사용하기 위한 약제의 제조에 있어 제23항 내지 제25항 중 어느 한 항의 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염의 용도. 26. Use of the N-acylhydrazone derivative of any one of claims 23 to 25, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the prevention or treatment of a cell proliferative disease.
  29. 제23항 내지 제25항 중 어느 한 항의 N-아실히드라존 유도체, 이의 입체이성질체 또는 이의 약학적으로 허용가능한 염을 필요로 하는 대상에 투여하는 것을 포함하는, 세포증식성 질환을 치료하는 방법.A method for treating a cell proliferative disease, comprising administering the N-acylhydrazone derivative of any one of claims 23 to 25, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof.
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