JP5479105B2 - Novel ubiquitin-binding small molecule - Google Patents

Description

  The present invention relates to a novel compound having ubiquitin-binding activity, a medicament such as an anticancer agent containing the compound as an active ingredient, a novel ubiquitin-binding small molecule, and the like.

Ubiquitin is a protein with a molecular weight of 8,500 consisting of 76 amino acids that is universally found in eukaryotic cells. The cell cycle is controlled by modifying the functions of these proteins by modifying the functions of the three enzymes, ubiquitin activating enzyme (E1), ubiquitin-binding enzyme (E2), and ubiquitin ligase (E3). Involved in various life activities such as DNA repair and signal transduction. In many cases, ubiquitin is added to ubiquitin modified with other proteins to form a polyubiquitin chain, and the polyubiquitin chain functions as a proteasome recognition signal, and the protein to which ubiquitin is added is degraded.
The proteasome plays an essential role in life activity by selectively degrading proteins in cooperation with ubiquitin which serves as a landmark for degrading target proteins. In recent years, various intractable diseases such as cancer, neurological diseases and immune diseases are considered to be caused by the breakdown of a proteolytic apparatus composed of proteasome and ubiquitin. In particular, the usefulness of proteasome function inhibition for cancer treatment has recently been highlighted, and is expected to become a target for new anticancer agents.

  From such a point of view, development of an anticancer agent that kills cancer cells by accumulating abnormal proteins by inhibiting the action of proteasomes in cancer cells has been performed. For example, 1) proteasome inhibitors such as bortezomib, lactacystin, epoxomicin, etc. 2) ubiquitin activating enzyme E1 inhibitors such as panepophenanthrin, 3) chlorofusine C ), Ubiquitin ligase E3 inhibitors such as Calcon C, and 4) ubiquitinase inhibitors such as Prostaglandin J2 are known. Bortezomib has already been approved by the FDA as a pharmaceutical product, but its side effects are relatively strong. Therefore, development of a compound that acts on the ubiquitin-proteasome system and has a different side of action from bortezomib or the like and has few side effects is desired.

  Ubiquitin 1 is known to have a function of binding to a protein modified with ubiquitin, that is, ubiquitinated and transporting the ubiquitinated protein to the proteasome. However, no compound having an ubiquitin inhibitory activity has been found. First, there are many unclear parts about the function of ubiquitin in the ubiquitin-proteasome system.

On the other hand, a tetracyclic coumarin compound having anticancer activity has been reported from Brazilian Hypericum grass (Patent Document 1, Non-Patent Document 1), but there is no disclosure or suggestion of the relationship with the ubiquitin-proteasome system. . In addition, antibacterial action, antifungal action, antiviral action, immunopotentiation action, immunomodulation action, etc. of compounds similar to such coumarin compounds are known. (Patent Document 2).
Republished WO 2004/066994 JP 2005-508283 A Kimura et al., Int J Cancer. 2005 Jan 1; 113 (1): 158-65

  An object of the present invention is to provide a compound that acts on the ubiquitin-proteasome system, in particular, a compound having a ubiquitin-binding action, preferably further having an anticancer action.

  The present inventors have discovered a tetracyclic coumarin compound (GUT-70) having an anticancer activity from Brazilian Hypericum grass (Non-patent Document 1). Chemical modification was performed using GUT-70 as a lead compound, various derivatives were synthesized, and their pharmacological action, particularly cancer cell growth inhibitory activity, was analyzed. The compound represented by the following general formula (I) was ubiquitin-binding activity. And ubiquitin-proteasome inhibitory activity. Based on this finding, the present inventors have conducted intensive studies and have completed the present invention.

That is, the present invention is as follows.
[1] The following general formula (I):

{In formula (I),
R ₁ represents hydrogen, an alkyl group, a cycloalkyl group, an alkenyl group which may have a substituent, an alkynyl group which may have a substituent, an aryl group which may have a substituent, a substituent A heterocyclic group which may have a group, an amino group which may have a substituent, an alkoxy group, an alkylcarbonyl group, an arylcarbonyl group which may have a substituent, and a substituent. May be a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, an alkoxycarbonyl group, an optionally substituted carboxyl group, an optionally substituted aminocarbonyl group or a general formula ( 1):

[In Formula (1),
R ₁₆ , R ₁₇ and R ₁₈ are the same or different and each represents hydrogen, an alkyl group, a cycloalkyl group, an aryl group which may have a substituent, a heterocyclic group which may have a substituent, or a dialkylamino group. Represents an alkenyl group which may have a substituent]
Means a structure represented by
R ₂ is hydrogen, an alkyl group, an aryl group which may have a substituent, a halogen atom, a cyano group, a cycloalkyl group, an alkylcarbonyl group, an arylcarbonyl group which may have a substituent, a carboxyl group , Alkoxycarbonyl group, thiol group, alkylthio group, arylthio group which may have a substituent, alkylsulfonyl group, arylsulfonyl group which may have a substituent, amino which may have a substituent Carbonyl group, optionally substituted aminosulfonyl group, optionally substituted aryloxycarbonyl group, amino group, alkylamino group, dialkylamino group, acylamino group, alkylsulfonylamino group or substituted An arylsulfonylamino group optionally having a group, or OR _2a (wherein R _2a is , Hydrogen, an alkyl group, an alkenyl group, an alkynyl group or an aryl group which may have a substituent, and OR _2a together with R ₁ is represented by the general formula (2):

(Wherein R ₁₉ and R ₂₀ are the same or different and each represents hydrogen, an alkyl group, a cycloalkyl group, or an aryl group which may have a substituent, or C—R ₁₉ R ₂₀ represents C ═O, R ₂₁ and R ₂₂ are the same or different and are each a hydrogen atom, a halogen atom, a cyano group, a hydroxy group, an alkyl group, a cycloalkyl group, an aryl group which may have a substituent, or an alkoxy group. , An alkylcarbonyl group, an arylcarbonyl group optionally having a substituent, a carboxyl group, an alkoxycarbonyl group, an amino group, an alkylamino group, a dialkylamino group or an acylamino group, and a broken line is condensed with a benzene ring Shows the part)
A ring represented by:
R ₃ represents hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, or an aryl group;
R ₄ represents hydrogen or OR _4a (wherein R _4a represents hydrogen, an alkyl group, an alkenyl group, an alkynyl group or an aryl group), and OR _4a is represented by the general formula (2) together with R _3. Can form a ring
formula:

The group represented by
Formula (a):

(Wherein X ₁ represents O, NH or NR ₁₅ (wherein R ₁₅ represents an alkyl group, a cycloalkyl group or an aryl group which may have a substituent), R _5a and R _5b is the same or different and is hydrogen, an alkyl group which may have a substituent, an aryl group which may have a substituent, a halogen atom, a cyano group, a hydroxy group, a cycloalkyl group, an alkoxy group, Alkylcarbonyl group, arylcarbonyl group optionally having substituent, carboxyl group, alkoxycarbonyl group, aryloxy group optionally having substituent, thiol group, alkylthio group, having substituent Arylthio group, alkylsulfonyl group, arylsulfonyl group which may have a substituent, aminocarbonyl group which may have a substituent, substituent An optionally substituted aminosulfonyl group, an optionally substituted aryloxycarbonyl group, an amino group, an alkylamino group, a dialkylamino group, an acylamino group, an alkylsulfonylamino group, or a substituent. Meaning a good arylsulfonylamino group), or
Formula (b):

(Wherein R ₆ and R ₇ are the same or different and each represents hydrogen or an alkyl group which may have a substituent, or C—R ₆ R ₇ represents C═O, and R ₈ represents Means an oxo group or an alkyl group;

Or a bond represented by a single bond or a double bond)
Formula (c):

(Wherein R ₉ represents hydrogen or an alkyl group, R ₁₀ represents hydrogen or an alkyl group, and R ₁₁ represents an arylcarbonyl group which may have a substituent). Or a group
Formula (d):

(Wherein X ₂ represents N or CH, and R ₁₂ represents hydrogen or an optionally substituted amino group) or a group represented by formula (e) :

(In the formula, R ₁₃ represents hydrogen or an aryl group which may have a substituent, and R ₁₄ represents hydrogen or an alkyl group which may have a substituent). A ubiquitin-binding small molecule comprising a compound represented by the group} or a pharmaceutically acceptable salt thereof as an active ingredient.
[2] The ubiquitin-binding small molecule according to [1] above, wherein the compound represented by the general formula (I) is a compound represented by the following general formula (II):

{In formula (II)
R _{1 ′} is hydrogen, a heterocyclic group, an optionally substituted amino group, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or the general formula (1 ′):

[In the formula (1 ′),
R _{16 ′} , R _{17 ′} and R _{18 ′} are the same or different and each represents hydrogen, an alkyl group or an aryl group which may have a substituent.
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents hydrogen or OR _{4a ′} (wherein R _{4a ′} represents an alkyl group), and OR _{4a ′} together with R _{3 ′ represents the} general formula (2 ′):

[Wherein R _{19 ′} and R _{20 ′} are the same or different and each represents an alkyl group, and the broken line represents a condensed portion with a benzene ring]
A ring represented by:
formula:

The group represented by
Formula (a ′):

Wherein R _{5a ′} and R _{5b ′} are the same or different and each represents hydrogen or an alkyl group which may have a substituent, or a group represented by formula (b ′) :

(Wherein R _{8 ′} represents an alkyl group).
[3] The compound represented by the general formula (I) is:
5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (GUT- 70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), or 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The ubiquitin-binding small molecule according to [1] above.
[4] The compound represented by the general formula (I) is:
5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (GUT- 70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), or Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The ubiquitin-binding small molecule according to [1] above.
[5] 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-, wherein the compound represented by the general formula (I) has the following structural formula The ubiquitin-binding small molecule according to the above [1], which is 2H-chromen-2-one.

[6] The ubiquitin-binding small molecule according to the above [1], which is used for prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases.
[6-1] Prevention or prevention of at least one disease selected from the group consisting of cancer, neurodegenerative disease, autoimmune disease and infectious disease, which contains the ubiquitin-binding small molecule according to [1] as an active ingredient Therapeutic agent.
[6-2] A cancer, a neurodegenerative disease, an autoimmune disease, comprising administering a therapeutically effective amount of the ubiquitin-binding small molecule according to the above [1] to a patient in need of the small molecule. A method for preventing or treating at least one disease selected from the group consisting of infectious diseases.
[7] A topoisomerase II inhibitor comprising as an active ingredient a compound represented by the general formula (I) (as defined above) or a pharmaceutically acceptable salt thereof.
[8] The topoisomerase II inhibitor according to [7] above, wherein the compound represented by the general formula (I) is a compound represented by the general formula (II) (as defined above).
[9] The compound represented by the general formula (I) is:
5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (GUT- 70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), or 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The topoisomerase II inhibitor according to [7] above.
[10] The compound represented by the general formula (I) is:
5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (GUT- 70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), or Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The topoisomerase II inhibitor according to [7] above.
[11] 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-, wherein the compound represented by the general formula (I) has the following structural formula The topoisomerase II inhibitor according to [7] above, which is 2H-chromen-2-one.

[12] The topoisomerase II inhibitor according to the above [7], which is used for the prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative disease, autoimmune disease and infectious disease.
[12-1] Prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases, containing the topoisomerase II inhibitor according to [7] above as an active ingredient Agent.
[12-2] Cancer, neurodegenerative disease, autoimmune disease and infection comprising administering to a patient in need of administration of a therapeutically effective amount of the topoisomerase II inhibitor according to [7] above A method for preventing or treating at least one disease selected from the group consisting of diseases.
[13] A compound represented by the general formula (I),
However, the following general formula (Ia):

{In formula (Ia),
R _{1 ″} is an alkyl group, a cycloalkyl group, an aryl group which may have a substituent, an alkoxy group, an alkylcarbonyl group, an arylcarbonyl group which may have a substituent, an alkoxycarbonyl group, a carboxyl Group or general formula (1a):

[In Formula (1a),
R _{16 ″} , R _{17 ″} and R _{18 ″} are the same or different and each represents hydrogen, an alkyl group, a cycloalkyl group or an aryl group which may have a substituent.
Means a structure represented by
R _{2 ″} is hydrogen, a halogen atom, a cyano group, an alkyl group, a cycloalkyl group, an aryl group that may have a substituent, an alkylcarbonyl group, an arylcarbonyl group that may have a substituent, Carboxyl group, alkoxycarbonyl group, thiol group, alkylthio group, arylthio group which may have a substituent, alkylsulfonyl group, arylsulfonyl group which may have a substituent, may have a substituent Good aminocarbonyl group, optionally substituted aminosulfonyl group, optionally substituted aryloxycarbonyl group, amino group, alkylamino group, dialkylamino group, acylamino group, alkylsulfonylamino group or optionally substituted aryl-sulfonylamino group, or oR 2a _'' (wherein, R _{2 ''} Is hydrogen, it means an aryl group which may have an alkyl group or a substituted group) means;
R ₁₉ and R ₂₀ are the same or different and each represents hydrogen, an alkyl group, a cycloalkyl group or an aryl group which may have a substituent, or C—R ₁₉ R ₂₀ represents C═O. R ₂₁ and R ₂₂ are the same or different and are each a hydrogen atom, a halogen atom, a cyano group, a hydroxy group, an alkyl group, a cycloalkyl group, an aryl group which may have a substituent, an alkoxy group, or an alkylcarbonyl group. Represents an optionally substituted arylcarbonyl group, carboxyl group, alkoxycarbonyl group, amino group, alkylamino group, dialkylamino group or acylamino group;
X ₁ represents O, NH or NR ₁₅ (wherein R ₁₅ represents an alkyl group, a cycloalkyl group or an aryl group which may have a substituent);
R _{5a ″} and R _{5b ″} are the same or different and each represents hydrogen, an alkyl group, an aryl group which may have a substituent, a halogen atom, a cyano group, a hydroxy group, a cycloalkyl group, an alkoxy group, an alkylcarbonyl Group, arylcarbonyl group optionally having substituent, carboxyl group, alkoxycarbonyl group, aryloxy group optionally having substituent, thiol group, alkylthio group, optionally having substituent An arylthio group, an alkylsulfonyl group, an arylsulfonyl group which may have a substituent, an aminocarbonyl group which may have a substituent, an aminosulfonyl group which may have a substituent, and a substituent; Aryloxycarbonyl group, amino group, alkylamino group, dialkylamino group, acylamino group, Have a Niruamino groups or substituents means aryl sulfonylamino group}
Or a compound represented by
An anticancer agent comprising a pharmaceutically acceptable salt as an active ingredient.
[13-1] A therapeutically effective amount of the compound represented by the general formula (I) (excluding the compound represented by the general formula (Ia)) or a pharmaceutically acceptable salt thereof, A method for preventing or treating cancer comprising administering to a patient in need thereof.
[14] The anticancer agent according to [13] above, wherein the compound represented by the general formula (I) is a compound represented by the general formula (II).
[14-1] The method according to [13-1] above, wherein the compound represented by the general formula (I) is a compound represented by the general formula (II).
[15] A compound represented by the general formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), or 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The anticancer agent according to [13] above.
[15-1] The compound represented by the general formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), or 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The method described in [13-1] above.
[16] The compound represented by the general formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), or Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The anticancer agent according to [13] above.
[16-1] The compound represented by formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), or Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The method described in [13-1] above.
[17] 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-, wherein the compound represented by the general formula (I) has the following structural formula: The anticancer agent according to [13] above, which is 2H-chromen-2-one.

[17-1] The compound represented by the general formula (I) has the following structural formula, and 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4- The method according to [13-1] above, which is propyl-2H-chromen-2-one.

[18] The anticancer agent according to any one of [13], [14], [15], [16] and [17], which has a binding activity to ubiquitin and exhibits an anticancer activity.
[18-1] The above [13-1], [14-1], [15-1], wherein the compound represented by the general formula (I) has a binding activity to ubiquitin and exhibits an anticancer activity. [16-1] The method according to any one of [17-1].
[19] Cancer is acute myeloid leukemia, acute lymphoblastic leukemia, malignant lymphoma, choriocarcinoma, multiple myeloma, soft tissue tumor, small cell lung cancer, chronic myelogenous leukemia, medullary thyroid cancer, osteosarcoma, head and neck [13], [14], [13] selected from the group consisting of cancer, esophageal cancer, non-small cell lung cancer, colon cancer, gastric cancer, biliary tract cancer, brain tumor, malignant melanoma, kidney cancer, pancreatic cancer, liver cancer [15] The anticancer agent according to any one of [16], [17] and [18].
[19-1] Cancer is acute myeloid leukemia, acute lymphocytic leukemia, malignant lymphoma, choriocarcinoma, multiple myeloma, soft tissue tumor, small cell lung cancer, chronic myelogenous leukemia, thyroid medullary cancer, osteosarcoma, [13-1], [13] selected from the group consisting of head and neck cancer, esophageal cancer, non-small cell lung cancer, colon cancer, stomach cancer, biliary tract cancer, brain tumor, malignant melanoma, kidney cancer, pancreatic cancer, liver cancer 14-1], [15-1], [16-1], [17-1], The method of any one of [18-1].
[20] A compound represented by the general formula (I),
However, the compound represented by the general formula (Ia) and the following general formula (Ib):

{In formula (Ib),
R _{1 ′ ″} means an alkyl group or an aryl group;
R _{2a ′ ″} means hydrogen, an alkyl group, an alkenyl group, an alkynyl group or an aryl group;
R _{3 ′ ″} means hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group or an aryl group;
R _{4a ′ ″} means hydrogen, an alkyl group, an alkenyl group, an alkynyl group or an aryl group [wherein OR _{4a ′ ″} represents a ring represented by the general formula (2) together with R _{3 ′ ″.} Can be formed];
R _{5a ′ ″} means an alkyl group, an aryl group or a hydroxy group}
Or a pharmaceutically acceptable salt thereof.
[21] The compound of the above-mentioned [20], wherein the compound represented by the general formula (I) is a compound represented by the general formula (II) or a pharmaceutically acceptable salt thereof (provided that R _{1 ′} Represents a structure represented by the general formula (1 ′), OR _{4a ′} does not form a ring represented by the general formula (2 ′) together with R _{3 ′} .
[22] The compound represented by the general formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), or 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The compound of the above-mentioned [20] or a pharmaceutically acceptable salt thereof.
[23] The compound represented by the general formula (I) is:
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), or Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
The compound of the above-mentioned [20] or a pharmaceutically acceptable salt thereof.
[24] 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-, wherein the compound represented by the general formula (I) has the following structural formula: The compound of the above-mentioned [20] or a pharmaceutically acceptable salt thereof which is 2H-chromen-2-one.

[25] A medicament comprising the compound according to any one of [20] to [24] above or a pharmaceutically acceptable salt thereof as an active ingredient.
[26] The medicament described in [25] above, which is an anticancer agent.
[27] A ubiquitin-binding small molecule or topoisomerase II inhibitor for the manufacture of a preventive or therapeutic agent for at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases use.
[28] Prevention or prevention of at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases, wherein the ubiquitin-binding small molecule or topoisomerase II inhibitor is used. A commercial package describing what can or should be used for treatment.
[29] Use of a compound represented by the general formula (I) for the manufacture of a prophylactic or therapeutic agent for at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases .
[30] Can the compound represented by the general formula (I) and the compound be used for prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative disease, autoimmune disease and infectious disease? Or a commercial package stating that it should be used.

  The present invention provides a novel compound having ubiquitin-binding activity and a medicament containing the compound as an active ingredient. Furthermore, the compound has a topoisomerase II inhibitory action. Due to these actions, the compound of the present invention is useful as a prophylactic / therapeutic agent for various diseases related to the ubiquitin-proteosome system and various diseases related to topoisomerase II activity, and particularly useful as an anticancer agent.

It is the figure (fluorescence microscope image) which showed the influence which acts on mitosis of BNS-22. It is the figure which showed the result of having carried out the DNA dyeing | staining of the cell which processed BNS-22, and analyzing the cell cycle by FACS. 2N represents a diploid cell and 4N represents a tetraploid cell. It is the figure which showed the cell growth inhibitory effect of BNS-22 (WST-8 method). The vertical axis represents the proportion of living cells as cell growth (%), and the horizontal axis represents the concentration (μM) of BNS-22. It is a figure (electrophoresis image) which shows the result of having measured the inhibitory activity of BNS-22 topoisomerase II in vitro. FIG. 6 shows the effect of BNS-22 on the decoupling of continuous circular kinetoplast DNA by topoisomerase II. It is a figure (electrophoresis image) which shows the result of having analyzed the protein couple | bonded with BNS-22 specifically. Ubiquitin 1 was confirmed as a protein that specifically binds to and co-precipitates with BNS-22-bound beads. It is a figure (electrophoresis-immunoblot image) which shows accumulation | storage induction | guidance | derivation of the intracellular ubiquitination protein by BNS-22 treatment. (A) It is the figure which showed typically the procedure of the pull-down assay using a GUT-70 binding bead. (B) It is a figure (electrophoresis image) which shows the result of having analyzed the protein couple | bonded specifically with GUT-70. (A) It is the figure which represented typically the interaction | combination effect | action of the free GUT-70 which acts on the interaction of the immobilized GUT-70 and ubiquitin 1, and the said interaction. (B) Coomassie brilliant blue (CBB) stained image of purified recombinant ubiquitin 1. (C) It is a figure (electrophoresis-immunoblot image) which shows the result of having analyzed the protein couple | bonded with GUT-70 specifically. Ubiquitin 1 was confirmed as a protein that specifically binds to and coprecipitates with GUT-70-bound beads. In the presence of free GUT-70, the interaction between GUT-bound beads and ubiquitin 1 was inhibited. (A) It is the schematic diagram which considered the influence which GUT-70 and MG132 exert on the ubiquitin proteasome system. (B) It is a figure (electrophoresis-immunoblot image) which shows the accumulation | storage induction | guidance | derivation of intracellular ubiquitinated protein by GUT-70 process.

Hereinafter, the definition of each symbol used in this specification will be described in detail.
“Halogen” in the present specification includes fluorine, chlorine, bromine and iodine.

The “alkyl group” in the present specification means a linear or branched alkyl group, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert- pentyl, hexyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl, heptyl, octyl and the like, among _C 1 -C ₆ alkyl group is preferable.
The term “alkyl group optionally having substituent (s)” in the present specification refers to an alkyl group optionally having 1 to 5, preferably 1 to 3 substituents at substitutable positions. means. Examples of such a substituent include a carboxyl group, a dialkylaminocarbonyl group (eg, dimethylaminocarbonyl), an optionally substituted heterocyclic carbonyl group (described later), and an optionally substituted group. An aryl group (described later), an alkyloxycarbonyl group (eg, methoxycarbonyl, ethoxycarbonyl), an alkenylcarbonyloxy group (eg, isobutenylcarbonyloxy) and the like can be mentioned.

The “alkenyl group” in the present specification is a linear or branched alkenyl group such as vinyl, allyl, propenyl, isopropenyl, butenyl, isobutenyl, but-3-en-1-yl, penta 4-en-1-yl, hexa-5-en-1-yl and the like to, among others _C 2 -C ₆ alkenyl groups are preferred.
In the present specification, the “optionally substituted alkenyl group” refers to an alkenyl group optionally having 1 to 5, preferably 1 to 3 substituents at substitutable positions. means. Examples of such a substituent include a hydroxy group, a dialkylamino group (described later), and an aryl group (described later) which may have a substituent.

The “alkynyl group” in the present specification is a linear or branched alkynyl group, for example, ethynyl, prop-2-yn-1-yl, but-3-yn-1-yl, penta-4 - yn-1-yl, hexa-5-yl and the like to, among others _C 2 -C ₆ alkynyl groups are preferred.
In the present specification, “optionally substituted alkynyl group” means an alkynyl group optionally having 1 to 5, preferably 1 to 3 substituents at substitutable positions. means. Examples of such a substituent include a hydroxy group.

Examples of the “cycloalkyl group” in the present specification include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like, and among them, a C ₃ -C ₆ cycloalkyl group is preferable.

Examples of the “aryl group” in the present specification include phenyl, naphthyl (eg, 1-naphthyl, 2-naphthyl), biphenylyl, anthryl, phenanthryl and the like, and among them, a C ₆ -C ₁₄ aryl group is preferable. , C _6-10 aryl groups are more preferable, for example, phenyl groups and the like are generally used.
In the present specification, the “aryl group optionally having substituent (s)” refers to an aryl group optionally having 1 to 5, preferably 1 to 3 substituents at substitutable positions. means. Examples of such a substituent include a halogen (eg, bromine, chlorine), a cyano group, a nitro group, a hydroxy group, an aminocarbonyl group, an acetyl group, a carboxyl group, an alkoxycarbonyl group (described later), and a substituent. Examples thereof include an amino group (described later), an alkoxy group (described later), and an alkyl group (described above) which may have a substituent.

Examples of the “alkoxy group” in the present specification include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, and tert-pentyloxy. Hexyloxy, 2-ethylbutoxy, etc., among which a C ₁ -C ₆ alkoxy group is preferred.

  Unless otherwise specified, the “heterocyclic group” in the present specification includes an aromatic heterocyclic group and a non-aromatic heterocyclic group.

Here, examples of the aromatic heterocyclic group include 4 to 7 members (preferably 5 or 5) containing 1 to 4 heteroatoms selected from an oxygen atom, a sulfur atom and a nitrogen atom in addition to a carbon atom as a ring constituent atom. 6-membered) monocyclic aromatic heterocyclic group and condensed aromatic heterocyclic group. Examples of the condensed aromatic heterocyclic group include a ring corresponding to the 4- to 7-membered monocyclic aromatic heterocyclic group and a 5- or 6-membered aromatic heterocyclic ring containing 1 or 2 nitrogen atoms. (Eg, pyrrole, imidazole, pyrazole, pyrazine, pyridine, pyrimidine) from a ring in which 1 or 2 selected from a 5-membered aromatic heterocycle containing one sulfur atom (eg, thiophene) and a benzene ring are condensed Examples include groups to be derived.
As the “aromatic heterocyclic group”, for example,
Furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, furazanyl, 1,2,3- Monocyclic such as thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl An aromatic heterocyclic group; and benzofuranyl, isobenzofuranyl, benzo [b] thienyl, indolyl, isoindolyl, 1H-indazolyl, benzimidazolyl, benzoxazolyl, benzo [d] isoxazolyl, benzothiazolyl, benzo [d] Sothiazolyl, 1H-benzotriazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, naphthyridinyl, purinyl, pteridinyl, carbazolyl, α-carbolinyl, β-carbolinyl, γ-carbolinyl, acridinyl, phenoxazinyl, phenothiazinyl, phenothiazinyl Phenoxathiinyl, thiantenyl, phenathidinyl, phenathidinyl, phenanthrolinyl, indolizinyl, pyrrolo [1,2-b] pyridazinyl, pyrazolo [1,5-a] pyridyl, imidazo [1,2-a] pyridyl, imidazo [ 1,5-a] pyridyl, imidazo [1,2-a] pyridazinyl, imidazo [1,2-a] pyrimidinyl, 1,2,4-triazolo [4,3-a] pyridyl, 1,2,4- bird Zorro [4,3-b] fused aromatic heterocyclic group pyridazinyl and the like;
Is mentioned.

Examples of the non-aromatic heterocyclic group include 4 to 7 members (preferably 5 or 6 members) containing 1 to 4 heteroatoms selected from oxygen atoms, sulfur atoms and nitrogen atoms in addition to carbon atoms as ring constituent atoms. ) Monocyclic non-aromatic heterocyclic group and condensed non-aromatic heterocyclic group. Examples of the condensed non-aromatic heterocyclic group include a ring corresponding to the 4- to 7-membered monocyclic non-aromatic heterocyclic group, and a 5- or 6-membered aromatic containing 1 or 2 nitrogen atoms. 1 or 2 rings selected from a heterocyclic ring (eg, pyrrole, imidazole, pyrazole, pyrazine, pyridine, pyrimidine), a 5-membered aromatic heterocyclic ring containing 1 sulfur atom (eg, thiophene) and a benzene ring Examples thereof include a group derived from a condensed ring and a group obtained by partial saturation of the group.
As the “non-aromatic heterocyclic group”, for example,
Monocyclic non-aromatic heterocyclic groups such as azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, tetrahydrofuryl, thiolanyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, piperidyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, piperazinyl, etc .; , Dihydroquinolyl, isochromenyl, chromenyl (2H-chromenyl, 4H-chromenyl), 1,2,3,4-tetrahydroisoquinolyl, 1,2,3,4-tetrahydroquinolyl, 2,3-dihydrobenzo Condensed non-aromatic heterocyclic groups such as furanyl, benzo [1,3] dioxolyl;
Is mentioned.

  The “heterocyclic group optionally having substituent (s)” in the present specification means a heterocyclic ring optionally having 1 to 5, preferably 1 to 3 substituents at substitutable positions. Means group. Examples of such a substituent include halogen (described above), alkyl group (described above), aralkyl group (eg, benzyl), heterocyclic group (described above), and alkoxy group (described later).

In the present specification, the “optionally substituted amino group” is an amino group (monoalkylamino group, simply alkylamino) monosubstituted by an alkyl group (preferably a C ₁ -C ₆ alkyl group). Or a di-substituted amino group (dialkylamino group) or an aryl group (preferably a C ₆ -C ₁₄ aryl group) which may have a substituent (eg, a halogenated alkyl group). A substituted amino group (monoarylamino group) or a disubstituted amino group (diarylamino group), an amino group substituted with an optionally substituted aminocarbonyl group (described later), an alkenylcarbonyl group ( Examples include an amino group substituted with a later-described amino group and an amino group substituted with a heterocyclic carbonyl group (described later). Examples of the halogenated alkyl group include a trifluoromethyl group.
Specific examples of the “optionally substituted amino group” include methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, sec-butylamino, tert-butylamino, pentylamino, isopentyl amino, neopentyl amino, tert- pentylamino, _C 1 -C ₆ alkylamino group such hexylamino, dimethylamino, diethylamino, etc. N- ethyl -N- methylamino di _(C 1 -C ₆₎ alkyl amino group, phenylamino, 1-naphthylamino, 2-naphthylamino, _C 6 _{-C 14} arylamino group such as trifluoroacetic phenylamino, diphenylamino, di _(C 6 _{-C 14)} aryl amino groups such as dinaphthylamino , Phenylaminocarbonyl, butenylcarbonyl Benzodioxolyl carbonyl, and the like.

  The “alkylamino group” and “dialkylamino group” in the present specification are the same as those exemplified as the “amino group optionally having substituent (s)” described above.

  In the present specification, “alkylcarbonyl group”, “alkenylcarbonyl group”, “alkoxycarbonyl group”, “arylcarbonyl group optionally having substituent”, “cycloalkylcarbonyl group” and “having substituent” The aminocarbonyl group which may optionally be represented by the above-mentioned “alkyl group”, “alkenyl group”, “alkoxy group”, “aryl group optionally having substituent”, “cycloalkyl group” and “ It means a carbonyl group substituted with an “amino group optionally having substituent (s)”.

The “alkylcarbonyl group” is preferably a C ₁ -C ₆ alkyl-carbonyl group, specifically, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 3-methylbutanoyl, 2,2 -Dimethylpropanoyl etc. are mentioned.
The “alkenylcarbonyl group” is preferably a C ₂ -C ₆ alkenyl-carbonyl group, specifically, vinylcarbonyl, 1-propenylcarbonyl, 2-propenylcarbonyl, 2-butenylcarbonyl, 3-butenylcarbonyl. Etc.
The “alkoxycarbonyl group” is preferably a C ₁ -C ₆ alkoxy-carbonyl group, specifically, methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl, isobutoxycarbonyl, and sec-butoxycarbonyl, tert-butoxycarbonyl and the like.
The “cycloalkylcarbonyl group” is preferably a C ₃ -C ₆ cycloalkyl-carbonyl group, specifically, cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, cycloheptylcarbonyl, cyclooctylcarbonyl and the like. Is mentioned.
The “arylcarbonyl group optionally having substituent (s)” is preferably a C ₆ -C ₁₄ aryl-carbonyl group, and specific examples include benzoyl, 1-naphthoyl, 2-naphthoyl and the like.
In the present specification, the “optionally substituted aminocarbonyl group” means, for example, an alkylamino group (preferably an amino group mono- or di-substituted with C ₁ -C ₆ alkyl) or an arylamino group (preferably an amino group mono- or di-substituted by C ₆ -C ₁₄ aryl group) refers to a carbonyl group substituted with, in particular, methylaminocarbonyl, ethylaminocarbonyl, n- propyl aminocarbonyl, isopropyl Aminocarbonyl, n-butylaminocarbonyl, isobutylaminocarbonyl, tert-butylaminocarbonyl, n-pentylaminocarbonyl, isopentylaminocarbonyl, hexylaminocarbonyl, dimethylaminocarbonyl, diethylaminocarbonyl, di-n-propylaminocarbonyl, di Sopropylaminocarbonyl, di-n-butylaminocarbonyl, diisobutylaminocarbonyl, ditert-butylaminocarbonyl, di-n-pentylaminocarbonyl, diisopentylaminocarbonyl, dihexylaminocarbonyl, phenylaminocarbonyl, naphthylaminocarbonyl, etc. It is done.

In the present specification, the “cycloalkenylcarbonyl group” means a carbonyl group substituted with a C ₃ -C ₆ cycloalkenyl group such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl and the like.

The “heterocyclic carbonyl group” in the present specification means a carbonyl group substituted with the above-mentioned “heterocyclic group”. Examples of the “heterocyclic group” of the “heterocyclic carbonyl group” include a ring corresponding to the aromatic or non-aromatic heterocyclic group exemplified as the heterocyclic group. Specific examples of the “heterocyclic carbonyl group” include benzofuranylcarbonyl, thienylcarbonyl, benzimidazolylcarbonyl, pyrimidinylcarbonyl, 1-pyrrolidinylcarbonyl, piperidinocarbonyl, 1-piperazinylcarbonyl, morpholinocarbonyl, thiomorpholino Examples include carbonyl and the like.
The “heterocyclic carbonyl group optionally having substituent (s)” in this specification means a heterocyclic ring optionally having 1 to 3 substituents at substitutable positions in addition to the carbonyl group. Means a carbonyl group. Examples of such a substituent include halogen (described above), alkyl group (described above), aralkyl group (eg, benzyl), heterocyclic group (described above), and alkoxy group (described later).

  Examples of the “substituent” in the “carboxyl group optionally having a substituent” in the present specification include an alkyl group (described above), an aryl group (described above), an aralkyl group (eg, benzyl, phenethyl) and the like. Can be mentioned.

In the present specification, the “alkylthio group” and the “arylthio group optionally having substituent (s)” are each substituted with the above-mentioned “alkyl group” and “aryl group optionally having substituent (s)”. Means a thio group.
The “alkylthio group” is preferably a C ₁ -C ₆ alkylthio group, and specific examples include methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, tert-butylthio and the like.
The “arylthio group optionally having substituent (s)” is preferably a C ₆ -C ₁₄ arylthio group optionally having substituent (s), and specific examples include phenylthio, naphthylthio and the like.

In the present specification, the “alkylsulfonyl group” and the “arylsulfonyl group optionally having substituent (s)” are substituted with the “alkyl group” and “aryl group optionally having substituent (s)”, respectively. Means a substituted sulfonyl group.
The “alkylsulfonyl group” is preferably a C ₁ -C ₆ alkylsulfonyl group, specifically, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, n-butylsulfonyl, isobutylsulfonyl, tert- And butylsulfonyl.
The “arylsulfonyl group optionally having substituent (s)” is preferably a C ₆ -C ₁₄ arylsulfonyl group optionally having substituent (s) such as phenylsulfonyl, naphthylsulfonyl and the like. Is mentioned.

Examples of the “optionally substituted aminosulfonyl group” include (C ₁ -C ₆ ) alkylamino-sulfonyl group, di (C ₁ -C ₆ ) alkylamino-sulfonyl group, and (C ₆ -C _14). ) Arylamino-sulfonyl group, di (C ₆ -C ₁₄ ) arylamino-sulfonyl group and the like, specifically, methylaminosulfonyl, ethylaminosulfonyl, n-propylaminosulfonyl, isopropylaminosulfonyl, n- Butylaminosulfonyl, isobutylaminosulfonyl, tert-butylaminosulfonyl, n-pentylaminosulfonyl, isopentylaminosulfonyl, hexylaminosulfonyl, dimethylaminosulfonyl, diethylaminosulfonyl, di-n-propylaminosulfonyl, diisopropylaminosulfur Examples include phonyl, di-n-butylaminosulfonyl, diisobutylaminosulfonyl, ditert-butylaminosulfonyl, di-n-pentylaminosulfonyl, diisopentylaminosulfonyl, dihexylaminosulfonyl, phenylaminosulfonyl, naphthylaminosulfonyl, and the like.

  Examples of the “aryloxy group which may have a substituent” in the present specification include phenoxy, 1-naphthyloxy, 2-naphthyloxy and the like.

  Examples of the “aryloxycarbonyl group which may have a substituent” in the present specification include phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxycarbonyl and the like.

  In the present specification, the “acylamino group” refers to —NHCOR ′ ″ (wherein R ′ ″ represents the “alkyl group” or the “aryl group optionally having substituent (s)”). Group), for example, acetylamino, propionylamino, butyrylamino, benzoylamino and the like.

  The “alkylsulfonylamino group” in the present specification means a sulfonylamino group substituted with the above “alkyl group”, specifically, for example, methylsulfonylamino, ethylsulfonylamino, n-propylsulfonylamino, isopropyl Examples include sulfonylamino, n-butylsulfonylamino, isobutylsulfonylamino, tert-butylsulfonylamino, n-pentylsulfonylamino, isopentylsulfonylamino, hexylsulfonylamino and the like.

  The “arylsulfonylamino group optionally having substituent (s)” in the present specification means a sulfonylamino group substituted with the above “aryl group optionally having substituent (s)”, specifically Includes phenylsulfonylamino, naphthylsulfonylamino and the like.

In formula (I), R ₁ is preferably (i) hydrogen, (ii) an alkenyl group which may have a substituent, (iii) an alkynyl group which may have a substituent, (iv) A heterocyclic group which may have a substituent, (v) an amino group which may have a substituent, (vi) an alkoxy group, (vii) an alkylcarbonyl group, and (viii) a substituent. An optionally substituted heterocyclic carbonyl group, (ix) a cycloalkylcarbonyl group, (x) a cycloalkenylcarbonyl group, (xi) an optionally substituted aminocarbonyl group, and (xii) a general formula (1) The structure represented is illustrated.

(Wherein the definitions of the symbols are as defined above)
In the formula (1), R ₁₆ is preferably hydrogen or an alkyl group, and R ₁₇ and R ₁₈ are preferably the same or different and each is an aryl group optionally having hydrogen, an alkyl group or a substituent. , A heterocyclic group which may have a substituent, a dialkylamino group or an alkenyl group which may have a substituent.

In formula (I), R ₂ is preferably hydrogen, a hydroxy group or an alkoxy group. Further _(wherein, the R _2a above synonymous) OR 2a as R ₂ are also preferred when they form a ring represented by which may have a substituent group together with R ₁ formula (2).

(Wherein the definitions of the symbols are as defined above)
In formula (2), R ₁₉ and R ₂₀ are preferably alkyl groups, and R ₂₁ and R ₂₂ are preferably hydrogen.
When OR _2a forms a ring represented by the general formula (2) which may have a substituent together with R ₁ , the compound represented by the formula (I) is a compound represented by the following general formula. is there.

(Wherein the definitions of the symbols are as defined above)
In formula (I), R ₃ is preferably hydrogen, an alkyl group or an alkoxy group.
In formula (I), R ₄ is preferably hydrogen, a hydroxy group or an alkoxy group.
Furthermore, OR _4a as R ₄ (wherein R _4a has the same _meaning as described above) is also preferred when forming a ring represented by the general formula (2) which may have a substituent together with R ₃ .

(Wherein the definitions of the symbols are as defined above)
In formula (2), R ₁₉ and R ₂₀ are preferably alkyl groups, and R ₂₁ and R ₂₂ are preferably hydrogen.
When OR _4a forms a ring represented by the general formula (2) which may have a substituent together with R ₃ , the compound represented by the formula (I) is a compound represented by the following general formula: is there.

(Wherein the definitions of the symbols are as defined above)

  In formula (I),

The group represented by these means groups represented by the following general formulas (a) to (e).

(Wherein the definitions of the symbols are as defined above)
In the formula (a), X ₁ is preferably O or NH, and R _5a and R _5b are preferably the same or different, each having hydrogen, an alkyl group which may have a substituent, or a substituent. It may be an aryl group.
In the formula (b), R ₆ and R ₇ are preferably the same or different and each represents an alkyl group. It is also preferred that C—R ₆ R ₇ forms C═O. R ₈ is preferably an oxo group or an alkyl group,

The bond represented by is a single bond or a double bond.
In formula (c), R ₉ is preferably hydrogen, R ₁₀ is preferably hydrogen, and R ₁₁ is preferably an arylcarbonyl group which may have a substituent.
In the formula (d), X ₂ is preferably N, and R ₁₂ is preferably an amino group which may have a substituent.
In the formula (e), R ₁₃ is preferably an aryl group, and R ₁₄ is preferably an alkyl group.

  Among the compounds represented by the formula (I), a compound represented by the following general formula (II) is particularly preferable.

{In formula (II)
R _{1 ′} is hydrogen, a heterocyclic group, an optionally substituted amino group, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or the general formula (1 ′):

[In the formula (1 ′),
R _{16 ′} , R _{17 ′} and R _{18 ′} are the same or different and each represents hydrogen, an alkyl group or an aryl group which may have a substituent.
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents hydrogen or OR _{4a ′} (wherein R _{4a ′} represents an alkyl group), and OR _{4a ′} together with R _{3 ′ represents the} general formula (2 ′):

[Wherein R _{19 ′} and R _{20 ′} are the same or different and each represents an alkyl group, and the broken line represents a condensed portion with a benzene ring]
A ring represented by:
formula:

The group represented by
Formula (a ′):

Wherein R _{5a ′} and R _{5b ′} are the same or different and each represents hydrogen or an alkyl group which may have a substituent, or a group represented by formula (b ′) :

(Wherein R _{8 ′} represents an alkyl group). }

As such a compound, for example,
5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (GUT- 70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38),
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44),
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate (BNS-45),
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64), and 3 Methyl [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
Preferably, 5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbute-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromene -8-on (GUT-70),
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-25),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61), and Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65)
And particularly preferably a compound having the following structural formula (8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one) It is.

  Among the compounds represented by the general formula (I), compounds not disclosed in Patent Document 1 and Patent Document 2 are novel compounds. That is, among the compounds represented by the general formula (I), the general formula (Ia):

(Wherein the definitions of the symbols are as defined above)
And a compound represented by the general formula (Ib):

(The definition of each symbol in the formula is as defined above.)
Compounds other than the compound represented by are novel compounds.

  In the present specification, a compound represented by the general formula (I) including a novel compound and a known compound may be referred to as a compound (I) or a compound of the present invention. Moreover, the compound (compound (II)) represented by general formula (II) which is a preferable compound is also within the range of the compound represented by general formula (I).

  Examples of the salt of the compound represented by the formula (I) include pharmaceutically acceptable salts, such as trifluoroacetic acid, acetic acid, lactic acid, succinic acid, maleic acid, tartaric acid, citric acid, gluconic acid, Acids with acids such as ascorbic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, cinnamic acid, fumaric acid, phosphonic acid, hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, sulfamic acid, sulfuric acid Addition salts; for example, metal salts such as sodium, potassium, magnesium, calcium; for example, salts with organic bases such as trimethylamine, triethylamine, pyridine, picoline, N-methylpyrrolidine, N-methylpiperidine, N-methylmorpholine, etc. Can be mentioned.

  When compound (I) has an isomer such as an optical isomer, a stereoisomer, a positional isomer, or a rotational isomer, any one isomer or a mixture of isomers is included in compound (I). Is done. For example, when compound (I) has an optical isomer, the optical isomer resolved from the racemate is also encompassed in compound (I). Each of these isomers can be obtained as a single product by a known synthesis method or separation method (concentration, solvent extraction, column chromatography, recrystallization, etc.).

  Compound (I) may be crystalline or amorphous. When the compound (I) is a crystal, it is included in the compound (I) regardless of whether it is a single crystal form or a mixture of crystal forms. The crystal can be produced by crystallization by applying a crystallization method known per se.

  Compound (I) may be a solvate (such as a hydrate) or a non-solvate, and both are encompassed in compound (I).

Compound (I) may be labeled with an isotope (eg, ³ H, ¹⁴ C, ³⁵ S, ¹²⁵ I, etc.) and the like.

The compound of the present invention has a ubiquitin binding activity (inhibits the function of the ubiquitin / proteasome system) and a topoisomerase II inhibitory action. Based on these actions, the compound of the present invention can be used in mammals (eg, humans, monkeys, cats, pigs, horses, cows, mice, rats, guinea pigs, dogs, rabbits, etc.) in the topoisomerase II and ubiquitin / proteasome systems. It is useful as a preventive or therapeutic agent for diseases related to onset and progression (diseases whose onset or progression is accelerated).
Examples of such diseases include cancer (eg, acute myeloid leukemia, acute lymphocytic leukemia, malignant lymphoma, choriocarcinoma, multiple myeloma, soft tissue tumor, small cell lung cancer, chronic myelogenous leukemia, medullary thyroid Cancer, osteosarcoma, head and neck cancer, esophageal cancer, non-small cell lung cancer, colon cancer, stomach cancer, biliary tract cancer, brain tumor, malignant melanoma, kidney cancer, pancreatic cancer, liver cancer), neurodegenerative diseases (eg, Alzheimer, Parkinson), autoimmune diseases (eg, rheumatoid arthritis, systemic lupus erythematosus), infectious diseases (eg, bacterial infections, viral infections such as AIDS) and the like.

  The content of the compound of the present invention in a medicament containing the compound of the present invention as an active ingredient (for example, an anticancer agent, etc.) is usually about 0.01 to about 99.9% by weight, preferably about 0.1 to about 99.9% by weight based on the whole preparation. About 50% by weight.

The dose of the compound of the present invention depends on age, body weight, general health condition, sex, meal, administration time, administration method, excretion rate, drug combination, and the degree of the medical condition being treated at the time of the patient, It is decided in consideration of these and other factors.
The dose varies depending on the target disease, symptom, administration subject, administration method, etc. For example, the daily dose of the compound of the present invention is about 0.1-100 mg / kg (body weight), preferably about 1-10 mg / kg (body weight) ), More preferably about 1 to 3 mg / kg (body weight) is preferably administered once or in 2 to 3 divided doses.

The compound of the present invention can be administered alone to obtain a desired effect, or can be used in appropriate combination with other anticancer agents, anti-inflammatory agents or immunotherapeutic agents.
Examples of other anticancer agents include antimetabolites (eg, methotrexate, 5-fluorouracil, etc.), alkylating agents (eg, cyclophosphamide, ifosfamide, etc.), platinum anticancer agents (eg, cisplatin, carboplatin, etc.), topoisomerase, etc. Inhibitors (eg, etoposide, etc.), anticancer antibiotics (eg, mitomycin, adriamycin, etc.), plant-derived anticancer agents (eg, vincristine, vindesine, taxol, etc.), tyrosine kinase inhibitors (eg, gefitinib, imatinib, etc.), Examples include humanized antibodies (eg, Herceptin).

  Anti-inflammatory agents include acetaminophen, phenacetin, ethenamide, sulpyrine, antipyrine, migrenin, aspirin, mefenamic acid, flufenamic acid, diclofenac sodium, loxoprofen sodium, phenylbutazone, indomethacin, ibuprofen, ketoprofen, naproxen, oxaprozin, flurbi Profen, fenbufen, pranoprofen, fructophenine, epilysole, tiaramid hydrochloride, zaltoprofen, gabexate mesilate, camostat mesilate, urinastatin, colchicine, probenade, sulfinpyrazone, benzbromarone, allopurinol, sodium sodium thiomalate, sodium hyaluronate , Sodium salicylate, morphine hydrochloride, salicylic acid, atropine Scopolamine, morphine, pethidine, levorphanol, ketoprofen, naproxen, non-steroidal anti-inflammatory agents such as oxymorphone or a salt thereof.

  Examples of immunotherapeutic agents include microorganisms or bacterial components (eg, muramyl dipeptide derivatives, picibanil, etc.), polysaccharides having immunopotentiating activity (eg, lentinan, schizophyllan, krestin, etc.), cytokines obtained by genetic engineering techniques (Eg, interferon, interleukin (IL), etc.), colony stimulating factor (eg, granulocyte colony stimulating factor, erythropoietin, etc.) and the like can be mentioned, among which IL-1, IL-2, IL-12 and the like are preferable.

The compound of the general formula (I) or a salt thereof is blended with a pharmaceutically acceptable carrier, solid preparations such as tablets, capsules, granules, powders; liquid preparations such as syrups and injections; It can be appropriately formulated as a transdermal absorption agent such as an ointment or a plaster; an inhalant; a suppository.
The medicament containing the compound of the present invention is administered orally or parenterally, and the above-mentioned compounds may be used alone or in combination of two or more.

  As the pharmaceutically acceptable carrier, various organic or inorganic carrier substances commonly used as pharmaceutical materials can be used. Specifically, excipients, lubricants, binders, disintegrants in solid preparations, solvents, dissolution aids, suspending agents, isotonic agents, buffering agents, soothing agents, etc. in liquid preparations can do. Moreover, formulation additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used as necessary.

  Examples of excipients include lactose, sucrose, glucose, starch, sucrose, microcrystalline cellulose, licorice powder, mannitol, sodium bicarbonate, calcium phosphate, calcium sulfate and the like.

  Examples of lubricants include magnesium stearate, stearic acid, calcium stearate, purified talc, colloidal silica and the like.

  Examples of the binder include crystalline cellulose, sucrose, mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone and the like.

  Examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium, carboxymethyl starch sodium and the like.

  Preferable examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like.

  Preferable examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

  Examples of suspending agents include, for example, surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate; polyvinyl alcohol, polyvinyl pyrrolidone, Examples include sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and the like.

  Preferable examples of the isotonic agent include sodium chloride, glycerin, D-mannitol and the like.

Preferable examples of the buffer include buffers such as phosphate, acetate, carbonate and citrate.
Preferable examples of the soothing agent include benzyl alcohol.

Preferable examples of the preservative include paraoxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.
Preferable examples of the antioxidant include sulfite and ascorbic acid.
Preferable examples of the colorant include tar pigment, caramel, iron sesquioxide, titanium oxide, riboflavin and the like.
Preferable examples of the sweetening agent include glucose, fructose, invert sugar, sorbitol, xylitol, glycerin, simple syrup and the like.

Production Method Hereinafter, the production method of the compound of the present invention will be described.
The compound of the present invention can be appropriately prepared by various methods, chemically synthesized, or obtained by separating it from an appropriate plant by extraction or the like and then appropriately modifying it as necessary. .
The compounds of the present invention include those which can be obtained by extraction from a suitable plant or can be obtained by derivatization therefrom. Examples of the plant include a plant of Gutiferae, from which 5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H , 8H-pyrano [2,3-f] chromen-8-one (GUT-70) can be obtained. A detailed extraction method is disclosed in Patent Document 1. Among the compounds represented by formula (I), the compound represented by formula (Ia) is disclosed in Patent Document 1 and the compound represented by Formula (Ib) is disclosed in Patent Document 2, respectively. Can be synthesized according to the literature.
Moreover, this invention compound can be manufactured by the method shown below or the method according to this.
In the following synthesis method, the starting compound may be used as a salt. As such a salt, those exemplified as the salt of the compound represented by the aforementioned formula (I) are used.
Unless otherwise stated, commercially available compounds can be easily obtained, or can be produced according to a method known per se or a method analogous thereto.
A protecting group may be used in each reaction, and as a method for removing the protecting group, a method known per se or a method analogous thereto is used. Specifically, a method of treating with acid, base, reduction, ultraviolet light, hydrazine, phenylhydrazine, sodium N-methyldithiocarbamate, tetrabutylammonium fluoride, palladium acetate or the like is used. The chemical properties of protecting groups, the method of their introduction, and their removal are described in detail, for example, in T. Greene and P. Wuts “Protective Groups in Organic Synthesis” (3 ^rd ed.), John Wiley & Sons NY (1999). ing.

Further, in each reaction and each reaction of starting compound synthesis, a generally known solvent may be used during the reaction.
Commonly known solvents include, for example, ethers such as tetrahydrofuran, diethyl ether, 1,2-dimethoxyethane, 1,4-dioxane, esters such as ethyl acetate and butyl acetate, and aromatic carbonization such as benzene and toluene. Aromatic heterocyclic compounds such as hydrogen, pyridine and lutidine, amides such as N, N-dimethylformamide and N-methylpyrrolidone, halides such as chloroform and methylene chloride, methanol, ethanol, 2-propanol, 2,2 -Alcohols such as dimethylethanol, hydrocarbon compounds such as hexane, heptane and petroleum ether, carboxylic acids such as formic acid and acetic acid, or water.
The solvent used in the reaction may be a single solvent or a mixture of two to six solvents.
Further, the reaction may be carried out in the presence of an amine such as triethylamine, N, N-diisopropylamine, pyridine, N-methylmorpholine, or a base such as sodium hydroxide or potassium carbonate.
The reaction may be carried out in the presence of an acid such as hydrochloric acid, sulfuric acid, or acetic acid.

(1) Synthesis of chroman and chromene derivatives In formula (I),

The group represented by
Formula (a ′)

(Wherein the definitions of the symbols are as defined above) or formula (b)

(Wherein the definitions of the symbols are as defined above)
A synthesis scheme of the compound is shown below (detailed reactions are according to the examples). In the scheme, specific groups may be described, but it will be apparent to those skilled in the art that alternative groups can be used.
Steps 1-3

In the formula, P is a hydroxy protecting group, and R, R ₁₀₀ and R ₅₀₀ are groups appropriately set according to the desired compound.
Examples of the hydroxy protecting group include a C ₁ -C ₆ alkyl group (eg, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl), a phenyl group, a trityl group, and a C ₇ -C ₁₀ aralkyl group (eg, Benzyl), formyl group, C ₁ -C ₆ alkyl-carbonyl group (eg, acetyl, propionyl), benzoyl group, C ₇ -C ₁₀ aralkyl-carbonyl group (eg, benzylcarbonyl), 2-tetrahydropyranyl group, 2 -Tetrahydrofuranyl group, silyl group (eg, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, tert-butyldimethylsilyl, tert-butyldiethylsilyl), C ₂ -C ₆ alkenyl group (eg, 1-allyl), etc. . These groups include halogen atoms (eg, fluorine, chlorine, bromine, iodine), C ₁ -C ₆ alkyl groups (eg, methyl, ethyl, propyl), C ₁ -C ₆ alkoxy groups (eg, methoxy, ethoxy, It may be substituted with 1 to 3 substituents selected from propoxy), nitro group and the like.
Step 1 is a step of introducing substituents (R and R ₁₀₀ ) and deprotecting a hydroxy protecting group. The reaction for introducing the substituent is appropriately selected according to the type of the substituent to be introduced.
Step 2 is a step of forming a chromene skeleton through a cyclization reaction.
Step 3 is a step of introducing a substituent (R ₁₀₀ ). The reaction for introducing the substituent is appropriately selected according to the type of the substituent to be introduced.
Any of these methods is carried out by a method commonly used in the art and a combination thereof.
Process 4-5

In the formula, P is a hydroxy protecting group (as defined above), and R ₁₀₀ , R ₃₀₀ , R ₄₀₀ and R ₅₀₀ are groups appropriately set according to the desired compound.
Step 4 is a step of introducing a hydroxy protecting group and introducing a substituent (R ₃₀₀ ), and step 5 is a step of deprotecting the hydroxy protecting group and removing the substituent (R ₃₀₀ ). Any of these methods is carried out by a method commonly used in the art and a combination thereof.
Step 6

In the formula, Me represents a methyl group, and R, R ₁₀₀ , R ₁₀₁ and R ₅₀₀ are groups appropriately set according to the desired compound.
Step 6 is a functional group substitution reaction. A reaction capable of substitution to a desired functional group is appropriately selected.
Step 7

R and R ′ are groups appropriately set according to the desired compound. Step 7 is a step of converting a carboxyl structure into a carbonyl structure, and is carried out by a method commonly used in the art and a combination thereof as appropriate.
Process 8-11

Me represents a methyl group, and R ₁₀₀ and R ₁₀₁ , and R ″ are groups appropriately set according to the desired compound. Steps 8 to 10 are all cyclization reactions, and a chromene or chroman derivative into which a desired substituent has been introduced is obtained. Step 11 is a step in which the compound obtained in Steps 8 to 10 is further subjected to a functional group substitution reaction or a reaction for introducing a substituent. Any reaction is appropriately set according to the desired compound, and is carried out by a method commonly used in the art and a combination thereof as appropriate.

(2) Synthesis of quinoline, isoquinoline, quinazoline derivative In the formula (I),

A group represented by formula (a ''):

(Wherein X _{1 ″} represents NH or NR ₁₅ (wherein R ₁₅ represents an alkyl group, a cycloalkyl group or an aryl group which may have a substituent), and other symbols. Is the same as defined above, or represents a group represented by formula (d):

(Wherein each symbol has the same meaning as above) or a group represented by formula (e):

(Wherein each symbol has the same meaning as described above), a synthesis scheme of a compound represented by the group shown below is shown (detailed reactions are according to the examples). In the scheme, there are cases where specific groups are described, but it is obvious to those skilled in the art that alternative groups may be used.
Step 1-4

In the formula, P and P ′ are the same or different and are a protecting group for a hydroxy group, and R ₁ has the same meaning as described above. Examples of the hydroxy-protecting group used in this step include the same as those described above.
Step 1 is a step of forming a quinoline skeleton by a cyclization reaction. Step 2 is a protecting group elimination reaction (deprotection). Step 3 is a reaction for introducing a substituent (R ₁ ). Step 4 is a step of forming a tricyclic structure having a quinoline skeleton by a cyclization reaction. Any of these reactions can be carried out by a reaction usually carried out in the art or by appropriately combining them.
Process 5

In the formula, the definition of each symbol is as defined above.
Step 5 is a reaction for converting a carboxyl group on the quinoline structure into an amide (amidation reaction). This reaction is also performed by a method commonly used in this field and a combination thereof as appropriate.
Step 6

In the formula, the definition of each symbol is as defined above.
Step 6 is a step of forming an isoquinoline skeleton by a cyclization reaction. This reaction is also performed by a method commonly used in this field and a combination thereof as appropriate.

  Hereinafter, the present invention will be described in detail with reference to examples and test examples. However, the present invention is not limited to these examples, and may be changed without departing from the scope of the present invention.

In addition, each symbol in an Example and a test example represents the following meaning.
AcOEt: ethyl acetate DMF: dimethylformamide DMAP: dimethylaminopyridine EtOH: ethanol Hex: hexane THF: tetrahydrofuran Me: methyl group WSCD: water-soluble carbodiimide (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide)
MQ: Milli-Q water (ultra pure water)
In this example, the physical property values were measured using the following apparatus.
Melting point measurement: B-545 from BUCHI Labortechnik AG
Elemental analysis: Yanagimoto mill MT-5 type (Duma method)
Mass spectrometry: JEOL SX-102 type (FAB: Fast Atom Bombardment method)

Example 1

The definition of each symbol in the formula is as defined above.
(Step 1-1)
223 mg of pyridin-4-yl- (2,4,6-trimethoxyphenyl) methanone and 436 mg of aluminum chloride were dissolved in 2 mL of nitrobenzene, 0.17 mL of butyroyl chloride was added, and the mixture was stirred at room temperature. After 11 hours, ice was added, 2N hydrochloric acid and saturated brine were added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate, and then the desiccant was filtered off and concentrated. The residue was purified by column chromatography (50 g of Wakogel C-200, developed with normal hexane: ethyl acetate = 4: 1 to 1: 1) to give 1- [2-hydroxy-4,6-dimethoxy-3- (pyridine-4) 100 mg of (carbonyl) phenyl] -butan-1-one was obtained as a pale yellow powder.

(Step 1-2)
1.00 g of 2,4,6-trimethoxynitrobenzene was dissolved in 10 mL of dichloromethane, and 4.7 mL of 1M boron tribromide / dichloromethane solution was added dropwise under argon atmosphere while cooling on a dry ice acetone bath. After stirring at the same temperature for 30 minutes, 2N hydrochloric acid was added, the temperature was raised to room temperature, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and the desiccant was filtered off and concentrated to obtain 931 mg of 3,5-dimethoxy-6-nitrophenol as a yellow powder. Similarly, 2-butyroyl-6-methoxyphenol was obtained.

(Step 1-3)
500 mg of 4,6-dimethoxy-salicylic acid, 503 mg of 1,2,3,4-tetrahydroisoquinoline and 970 mg of WSCD hydrochloride were dissolved in 10 mL of THF, 1 drop of DMF was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into ice, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off and concentrated to give (3,4-dihydro-2H-quinoline-1- Yl)-(2-hydroxy-4,6-dimethoxyphenyl) methanone was obtained as a pale yellow amorphous substance (541 mg).
The target compound 3,4-dihydro-2H-quinolin-1-yl)-(2,4,6-trimethoxyphenyl) methanone (BNS-58) was obtained in the same manner.
In the same manner, (2,3-dihydro-1H-indo-1-yl)-(2-hydroxy-4,6-dimethoxyphenyl) methanone, (3,4-dihydroisoquinolin-2 (1H) -yl )-(2-hydroxy-4,6-dimethoxyphenyl) methanone, (4-benzylpiperidin-1-yl)-(2-hydroxy-4,6-dimethoxyphenyl) methanone, 8-[(1H-1,2, , 3-Benzotriazol-1-yloxy) carbonyl] -7-hydroxy-4-propyl-2H-chromen-2-one, (1-piperidyl)-(2-hydroxy-4,6-dimethoxyphenyl) methanone It was. These were used in step 2-2 without further purification.

(Step 2-1)
100 mL of methanesulfonic acid (manufactured by Nacalai Tesque) was added to 20.0 g of phloroglucinol (manufactured by Tokyo Chemical Industry) and 25.3 g of ethyl 3-ketohexanoate (manufactured by Tokyo Chemical Industry), and the mixture was stirred at room temperature for 12 hours. The reaction mixture was poured into 100 mL of ice water and extracted with 150 mL of ethyl acetate. The organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate for 10 minutes. After the desiccant was filtered off, the solvent was distilled off under reduced pressure using a rotary evaporator. Under ice-cooling, normal hexane was added to the residue and stirred, and then the precipitate was collected by filtration and dried to obtain 35.3 g of 5,7-dihydro-4-propyl-2H-chromen-2-one as a white powder. . In a similar manner, ethyl 4- (5,7-dihydroxy-2-oxo-2H-chromen-4-yl) butanoate, 4- (3-bromophenyl) -5,7-dihydroxychromen-2-one, Ethyl 3- (5,7-dihydroxy-2-oxo-2H-chromen-4-yl) propanoate, 7-hydroxy-2-oxo-2H-chromene-8-carboxylic acid, and 7-hydroxy-2-oxo -2H-chromene-8-carboxylate methyl was obtained.

(Process 2-2)
15 mL of methanesulfonic acid was added to 2.00 g of 2-hydroxy-4,6-dimethoxybenzoic acid and 1.68 g of ethyl 3-ketohexanoate and stirred at room temperature. After 1 hour, the mixture was heated and stirred on an oil bath, and then allowed to cool to room temperature after 16 hours. After pouring into ice and extracting with ethyl acetate, the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated to obtain 1.45 g of 5,7-dimethoxy-4-propylcoumarin. Obtained.
In the same manner, 5,7-dimethoxy-4-propyl-2H-chromen-2-one, 4- (4-chlorophenyl) -5,7-dimethoxy-2H-chromen-2-one, 5,7-dimethoxy- 8-nitro-4-propyl-2H-chromen-2-one, 7-hydroxy-8-nitro-4-propyl-2H-chromen-2-one, and the target compound,
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-22),
5,7-dimethoxy-8- (1-methylpiperidin-4-yl) -4-propyl-2H-chromen-2-one (BNS-32),
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-51),
8- (3,4-dihydroisoquinolin-2 (1H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-52),
8-[(4-benzylpiperidin-1-yl) carbonyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-53),
5,7-dimethoxy-8- (piperidin-1-ylcarbonyl) -4-propyl-2H-chromen-2-one (BNS-54),
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-61),
Methyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate (BNS-65) is obtained. It was.

(Step 2-3)
640 mg of 2-butyroyl-6-methoxyphenol, 1.49 g of [(ethoxycarbonyl) methylene] triphenylphosphorane, and 5 mL of N, N-diethylaniline were heated and stirred on an 170 ° C. oil bath in an argon atmosphere. After 2 hours, the mixture was allowed to cool at room temperature, ethyl acetate was added, and the organic layer was washed with 2N hydrochloric acid, water and saturated brine in this order. After adding anhydrous sodium sulfate and drying, the desiccant was filtered and concentrated, and the resulting residue was purified by column chromatography (Wakogel C-200: 30 g, developed in normal hexane: ethyl acetate = 4: 1 to 1: 1), 595 mg of 8-methoxy-4-propyl-2H-chromen-2-one (BNS-17) as the target compound was obtained as a pale yellow crude powder. A part was recrystallized from benzene-normal hexane (melting point: 101 ° C.).
The target compound, 8-isonicotinoyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-23), was obtained in the same manner.

(Step 3-1)
122 mL of nitrobenzene was added to 20 g of 5,7-dihydroxy-4-propylcoumarin and 48.4 g of aluminum chloride, and the mixture was stirred at room temperature for 10 minutes until it became a homogeneous solution, followed by ice cooling. A solution prepared by dissolving 16.4 g of 2-methylbutanoic acid chloride (manufactured by Kanto Chemical Co., Ltd.) in 40 mL of carbon disulfide was added dropwise with care so that the internal temperature did not exceed 10 ° C. After completion of the dropwise addition, the mixture was heated and stirred on an oil bath preheated to 75 ° C. After 1.5 hours, the reaction solution was poured onto 500 g of ice, and 1 L of 2N hydrochloric acid was added while stirring. The mixture was extracted with ethyl acetate (1 L × 3 times), and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated on a rotary evaporator. Concentration was stopped when a precipitate began to appear, and normal hexane was added to the residue and stirred. Insoluble matter was collected by filtration, washed with normal hexane and dried to obtain 13.2 g of 5,7-dihydroxy-8- (2-methylbutyryl) -4-propyl-2H-chromen-2-one as a pale yellow powder. It was. In a similar manner, 8-acetyl-5,7-dihydroxy-4-propyl-2H-chromen-2-one, 5,7-dihydroxy-8-pentanoyl-4-propyl-2H-chromen-2-one, 5, 7-dihydroxy-8-propionyl-4-propyl-2H-chromen-2-one, 5,7-dihydroxy-8-isobutyryl-4-propyl-2H-chromen-2-one, 8-cyclobutylcarbonyl-5 7-dihydroxy-4-propyl-2H-chromen-2-one, 8-cyclohexylcarbonyl-5,7-dihydroxy-4-propyl-2H-chromen-2-one, 8-cyclopentylcarbonyl-5,7-dihydroxy- 4-propyl-2H-chromen-2-one, 5,7-dihydroxy-[(2E) -2-methylbut-2-noyl]- - give propyl -2H- chromen-2-one.

The definition of each symbol in the formula is as defined above.
(Step 4-1)
Mix 8.7 g of 5,7-dihydroxy-8- (2-methylbutyryl) -4-propyl-2H-chromen-2-one, 12.8 g of 4,4-dimethoxy-2-methyl-2-butanol and 17 mL of pyridine. The mixture was heated and stirred for 14 hours on an oil bath at 160 ° C. in an argon atmosphere. After allowing to cool at room temperature, Sm-2 (4,4-dimethoxy-2-methyl-2-butanol) and pyridine were distilled off on a rotary evaporator and dissolved in 100 mL of ethyl acetate. The organic layer was dissolved in 2N hydrochloric acid (50 mL × twice), The extract was washed with saturated brine (50 mL × 1) and then dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated, and the residue was purified by column chromatography (350 g of Wakogel C-200 (manufactured by Wako Pure Chemical Industries), developed with normal hexane: ethyl acetate = 10: 1 solution) to obtain 7.74 g as a pale yellow solid It was. 300 mg is recrystallized from petroleum ether (manufactured by Nacalai Tesque, containing 60-70% normal hexane) and dried under reduced pressure on diphosphorus pentachloride to give 5-hydroxy-2,2-dimethyl-6-6 which is the target compound. 249 mg of (2-methylbutanoyl) -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-2) was obtained as pale yellow crystals (melting point: 97-98 ° C.). .
In the same manner, the target compound is
5-hydroxy-2,2-dimethyl-6-propionyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-4),
6-acetyl-5-hydroxy-2,2-dimethyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-12),
5-hydroxy-2,2-dimethyl-6-pentanoyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-13),
5-hydroxy-2,2-dimethyl-6-isobutyryl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-14),
10- (3-bromophenyl) -5-hydroxy-2,2-dimethyl-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-15),
3- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-10-yl) propionic acid ethyl ester (BNS-16),
5-hydroxy-8,8-dimethyl-4-propyl-2H, 8H-pyrano [2,3-f] chromen-2-one (BNS-19),
5-Hydroxy-2,2-dimethyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-20) was obtained.
Similarly, 6-cyclobutylcarbonyl-5-hydroxy-2,2-dimethyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one, 6-cyclohexylcarbonyl-5 -Hydroxy-2,2-dimethyl-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one, 6-cyclopentylcarbonyl-5-hydroxy-2,2-dimethyl-10-propyl- 2H, 8H-pyrano [2,3-f] chromen-8-one was obtained.

(Step 4-2)
5-hydroxy-2,2-dimethyl-6- (2-methylbutanoyl) -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one 7.74 g, potassium carbonate 17.3 g Was suspended in 100 mL of acetone, 4.60 g of iodomethane was added, and the mixture was refluxed with heating on an oil bath for 2 hours. After cooling to room temperature, the mixture was filtered using Celite (No. 545, manufactured by Wako Pure Chemical Industries), and the organic layer was concentrated to obtain 7.50 g as a pale yellow solid. 300 mg was purified by column chromatography (Wakogel 200 45 g, developed in normal hexane: ethyl acetate = 4: 1 to 1: 1), and the resulting crude crystals were recrystallized from petroleum ether and dried under reduced pressure on diphosphorus pentachloride. 5-methoxy-2,2-dimethyl-6- (2-methylbutanoyl) -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-1) which is the target compound Was obtained as pale yellow crystals (melting point: 89-90 ° C.).
In the same manner, 5,7-dimethoxy-4-propyl-2H-chromen-2-one, 8-acetyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one, 8-acetyl-5, 7-diisopropoxy-4-propyl-2H-chromen-2-one, 7-methoxy-8-nitro-4-propyl-2H-chromen-2-one, and the target compound,
5,7-dimethoxy-8-propionyl-4-propyl-2H-chromen-2-one (BNS-5),
5-Methoxy-2,2-dimethyl-10- (3-oxo-3-pyrrolidin-1-ylpropyl) -6-propionyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS -24),
6-cyclobutylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-26),
6-cyclohexylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-27),
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-28),
6-cyclopentylcarbonyl-2,2-dimethyl-5-methoxy-10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-29),
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-35),
7-methoxy-8-[(2E) -2-methylbute-2-noyl] -4-propyl-2H-chromen-2-one (BNS-36),
8-acetyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-59),
8-[(1H-1,2,3-benzotriazol-1-yloxy) carbonyl] -7-methoxy-4-propyl-2H-chromen-2-one (BNS-60) was obtained.

(Process 5)
0.40 g of magnesium, 2 g of iodine, 5 mL of dehydrated diethyl ether (manufactured by Kanto Chemical) and 10 mL of dehydrated benzene (manufactured by Kanto Chemical) were mixed and heated to reflux for 45 minutes under an argon atmosphere. The obtained magnesium iodide solution was added 5-methoxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f. ] 309 mg of chromen-8-one (GUT-70) was dropped into a solution of 10 mL of benzene and heated to reflux. After 30 minutes, the mixture was allowed to cool to room temperature, 2N hydrochloric acid was added to dissolve the precipitate, and the mixture was extracted with diethyl ether (× 2 times). The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the desiccant was filtered off and concentrated. The obtained residue is purified by column chromatography (50 g of Wakogel C-200, developed with normal hexane: ethyl acetate = 10: 1 to 4: 1), and the resulting crude crystals are recrystallized from petroleum ether to obtain the target compound. 5-Hydroxy-2,2-dimethyl-6-[(2E) -2-methylbut-2-noyl] -10-propyl-2H, 8H-pyrano [2,3-f] chromen-8-one (BNS- 125 mg of 3) was obtained as pale yellow crystals (melting point: 143-145 ° C.).

The definition of each symbol in the formula is as defined above.
(Step 6-1)
7-Methoxy-8-nitro-4-propyl-2H-chromen-2-one 0.95 g was suspended in a mixed solution of 3 mL of acetic acid and 3.5 mL of concentrated hydrochloric acid, and tin (II) chloride dihydrate was stirred. 2.50 g was added. After stirring overnight at room temperature, the reaction mixture was poured into 2N aqueous sodium hydroxide solution, extracted with ethyl acetate, the organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated, and the residue was recrystallized from diisopropyl ether to obtain 592 mg of 8-amino-7-methoxy-4-propyl-2H-chromen-2-one as pale yellow crystals. In the same manner, 8-amino-5,7-dimethoxy-4-propyl-2H-chromen-2-one was obtained.

(Step 6-2)
200 mg of 8-amino-7-methoxy-4-propyl-2H-chromen-2-one was dissolved in 20 mL of ethyl orthoformate, one drop of concentrated sulfuric acid was added, and the mixture was heated to reflux on an oil bath. After confirming disappearance of the raw materials, the solvent was distilled off, 5 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for 1 hour and then carefully neutralized. Weak weak basicity (around pH 8) was confirmed, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated to obtain 185 mg of 8- (N-methylamino) -7-methoxy-4-propyl-2H-chromen-2-one as a light brown powder.

(Step 6-3)
8- (N-methylamino) -7-methoxy-4-propyl-2H-chromen-2-one (100 mg) and benzoyl chloride (63 mg) were dissolved in dichloromethane. Under ice-cooling, DMAP and then triethylamine were added and stirred for 30 minutes. After stirring for another 30 minutes at room temperature, water was added. Extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated after drying, the residue was purified by column chromatography (RediSep column 12 g, using combiflash, normal hexane only to 50 94 mg of white amorphous was obtained by applying a gradient to% ethyl acetate-normal hexane). By recrystallization from diisopropyl ether, the target compound, N- (7-methoxy-2-oxo-4-propyl-2H-chromen-8-yl) -N-methylbenzamide (BNS-41), was obtained as white crystals. As a product (melting point: 129.5 ° C.).
In a similar manner, the target compound is
(2E) -N- (7-methoxy-2-oxo-4-propyl-2H-chromen-8-yl) -N, 2-dimethylbut-2-amide (BNS-42),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide (BNS-55),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide (BNS-56),
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -N-methyl-1,3-benzodioxole-5-carboxamide (BNS-57) was obtained. .

(Step 6-4)
Tetrahydrofuran suspension solution of 100 mg of 8-amino-7-methoxy-4-propyl-2H-chromen-2-one under argon atmosphere, sodium hydride (21 mg of oil suspension was washed three times with normal hexane and dried under reduced pressure) The mixture was added with ice cooling and stirred for 1 hour. After adding 56 mg of phenyl isocyanate and stirring for 30 minutes, the mixture was warmed to room temperature and stirred for 1 hour. 1N Hydrochloric acid was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine. The extract was dried over anhydrous sodium sulfate, and the desiccant was filtered off and concentrated. The residue was purified by column chromatography (RediSep column 12 g, combined flash was used, and ethyl acetate-normal hexane was gradient from 25% to 50%), and the resulting crude powder was recrystallized from ethanol. N- (7-methoxy-2-oxo-4-propyl-2H-chromen-8-yl) -N′-phenylurea, which is the target compound, is obtained by recrystallizing the obtained crude crystals from benzene-petroleum ether. 24 mg of (BNS-43) was obtained as a pale yellow powder (melting point: 124 ° C.).

(Step 6-5)
0.18 g of 8-acetyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one was dissolved in 5 mL of EtOH, 0.066 g of benzaldehyde and 0.40 g of potassium hydroxide were added, and the mixture was stirred at room temperature for 2.5 days. Later, the solution was concentrated. Ethyl acetate was added and washed with saturated aqueous sodium hydrogen carbonate. HCl was added to the aqueous layer to make it acidic, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate and concentrated to obtain 0.22 g of a bright yellow powder. The obtained powder, 0.92 g of acetic anhydride and 0.43 g of sodium acetate were dissolved in benzene (20 mL) and heated under reflux for 1.5 hours. The reaction mixture was poured into water, stirred at room temperature for 3 hours, and then ethyl acetate. Extracted with. The extract was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue is purified by column chromatography (AcOEt / Hex = 1/1), which is the target compound, 5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl- 0.19 g of 2H-chromen-2-one (BNS-25) was obtained as a pale yellow powder.
In the same manner, the target compound is
5,7-dimethoxy-8-[(2E) -2-methyl-3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-30),
5,7-Dimethoxy-8-[(2E) -3- (1-methyl-1H-pyrrol-2-yl) prop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-31) ),
5,7-dimethoxy-8-[(2E, 4E) -5- (2-methoxyphenyl) penta-2,4-dienoyl] -4-propyl-2H-chromen-2-one (BNS-39),
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one (BNS-62),
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-64),
8-[(2E) -3- (2,3-dihydro-1,4-benzodioxin-6-yl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromene-2 -One (BNS-66), and 8-[(2E) -3- (9-ethyl-9H-carbazol-3-yl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H -Chromen-2-one (BNS-67) was obtained.

(Step 6-6)
To 0.30 g of 8-acetyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one was added 0.74 g of N, N-dimethylformamide dimethylacetal, and the mixture was heated and stirred at 85 ° C. overnight. The solvent was distilled off under reduced pressure, followed by column chromatography purification (NH silica, AcOEt), and the target compound, 8-[(2E) -3- (dimethylamino) prop-2-noyl] -5,7-dimethoxy-4. 0.17 g of propyl-2H-chromen-2-one (BNS-37) was obtained as a yellow powder (melting point: 207 ° C.).

(Step 6-7)
8-[(2E) -3- (dimethylamino) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one 200 mg, acetamidine hydrochloride 110 mg, potassium carbonate 160 mg The mixture was added to 3 mL of DMF and heated and stirred at 120 ° C. for 1 hour. After allowing to cool to room temperature, water was added and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. By purifying the residue by column chromatography (NH silica, AcOEt), the target compound, 5,7-dimethoxy-8- (2-methylpyrimidin-4-yl) -4-propyl-2H-chromen-2-one 50 mg of (BNS-40) was obtained as a pale yellow powder (melting point: 216 ° C.). In the same manner, 8- [2- (diethylamino) pyrimidin-4-yl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one was obtained.

(Step 6-8)
0.58 g of 8-acetyl-5,7-dimethoxy-4-propyl-2H-chromen-2-one was dissolved in 15 mL of ethyl acetate, 0.89 g of copper (II) bromide was added, and the mixture was heated to reflux for 2 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated and washed with diisopropyl ether. The resulting powder was suspended in chloroform and filtered. The filtrate was concentrated to obtain 0.58 g of white powder. The obtained powder was added to 2 mL of formamide, heated and stirred on an oil bath at 185 ° C. for 45 minutes, allowed to cool to room temperature, and saturated aqueous sodium hydrogen carbonate was added. The mixture was extracted with ethyl acetate, washed with water and then saturated brine. The organic layer is dried over anhydrous magnesium sulfate and concentrated, and the resulting residue is purified by column chromatography (development with NH silica, 5% methanol-chloroform) to give the target compound, 8- (1H-imidazole-4 -Yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-38) was obtained as an ocher powder.

(Step 6-9)
4 g of 5,7-dimethoxy-4-propyl-2H-chromen-2-one was dissolved in 40 mL of dichloromethane, and 3.93 g of iodine monochloride / dichloromethane solution was added dropwise thereto, followed by stirring at room temperature for 2 hours. After adding an aqueous sodium thiosulfate solution, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by column chromatography (AcOEt / Hex = 1/2) to obtain 2.68 g of a pale yellow powder. 0.29 g of the obtained powder, 0.50 g of 2-methyl-3-buten-2-ol, 0.85 mL of tetra-normal butylammonium bromide, 0.28 g of sodium carbonate and 18 mg of palladium acetate were dissolved in 4 mL of DMF, and the atmosphere was argon. The mixture was heated and stirred at 90 ° C. for 40 hours. The reaction mixture was filtered through celite, water was added to the filtrate, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, concentrated, and the residue was purified by column chromatography (AcOEt / Hex = 3/2). ) To obtain the target compound 8-[(1E) -3-hydroxy-3-methyl-1-buten-1-yl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-). 49) was obtained as a brown powder (melting point: 168 ° C.).

(Step 6-10)
Dissolve 0.5 g of 8-iodo-5,7-dimethoxy-4-propyl-2H-chromen-2-one and 0.14 g of 2-methyl-3-butyn-2-ol in 4 mL of DMF to obtain dichlorobistriphenylphosphine. 38 mg of palladium (II), 5 mg of copper iodide and 1 mL of triethylamine were added, and the mixture was stirred overnight on an oil bath at 55 ° C. under an argon atmosphere. The mixture was allowed to cool to room temperature, 1N hydrochloric acid was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and dried over anhydrous magnesium sulfate. The desiccant was filtered off, concentrated, and purified by column chromatography (Wakogel C-200, ethyl acetate: normal hexane = 1: 1), and the target compound, 8- (3-hydroxy-3-methyl-1-butyne- 0.20 g of 1-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one (BNS-50) was obtained as a brown powder (melting point: 179 ° C.).

(Step 6-11)
0.074 g of 8- [2- (diethylamino) pyrimidin-4-yl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one was suspended in 5 mL of ethanol, and 196 μL of 1N ethanol ethanol was added with stirring. It was. After stirring at room temperature for 1 hour and concentrating, the residue was dissolved in methanol and reprecipitated with diisopropyl ether under ice cooling. 8- [2- (Diethylamino) pyrimidin-4-yl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one hydrochloride, which is the target compound, is collected by filtration and dried under reduced pressure. 0.054 g of (BNS-63) was obtained as pale yellow crystals.

The definition of each symbol is as defined above.
(Step 7-1)
4- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-10-yl) butanoic acid methyl ester 260 mg was dissolved in methanol 20 mL. 2 mL of 10% aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2.5 hours. Ice water was added to the reaction solution, acidified with 10% aqueous hydrochloric acid solution, and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO ₄ ) and concentrated under reduced pressure. The obtained crude crystals were recrystallized from ethyl acetate to give 4- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3- f] 131 mg of chromen-10-yl) butanoic acid (BNS-6) was obtained as pale yellow crystals (melting point: 190-192 ° C.). 3- (5-Hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-10-yl) propionic acid, which is the target compound in the same manner (BNS-18) was obtained.

(Step 7-2)
4- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-10-yl) butanoic acid 300 mg was dissolved in dimethylformamide 10 mL, WSCD hydrochloride 165 mg and 1-hydroxybenzotriazole 143 mg were added and stirred for 15 minutes. 76 mg of dimethylamine hydrochloride and 236 mg of triethylamine were added and stirred at room temperature overnight. Ice water was added to the reaction solution, acidified with 10% aqueous hydrochloric acid solution, and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO ₄ ), and concentrated to dryness under reduced pressure. The obtained crude crystals were washed with ether and dried. The target compound, 4- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [2,3-f] chromen-10-yl) -N, N-dimethyl 187 mg of butanamide (BNS-8) was obtained as pale yellow crystals (melting point: 162-163 ° C.). In the same manner, the target compound, 5-hydroxy-2,2-dimethyl-10- [4-oxo-4- (4-pyridin-2-ylpiperazin-1-yl) butyl] -6-propionyl- 2H, 8H-pyrano [2,3-f] chromen-8-one (BNS-9), 3- (5-hydroxy-2,2-dimethyl-8-oxo-6-propionyl-2H, 8H-pyrano [ 2,3-f] chromen-10-yl) -N, N-dimethylpropanamide (BNS-21) was obtained.
In the same manner, 5-hydroxy-2,2-dimethyl-10- (3-oxo-3-pyrrolidin-1-ylpropyl) -6-propionyl-2H-pyrano [2,3-f] chromene-8 -Obtained ON.

The definition of each symbol is as defined above.
(Process 8)
To a solution of 1.43 g of 2-acetyl-3,5-dimethoxyphenol in 4.77 mL of methyl butanoate, 0.92 g of sodium hydride (70%, oil suspension) was carefully added and stirred at room temperature. After confirming disappearance of the raw material, ice water was added to crush excess sodium hydride, the aqueous layer was washed with ethyl acetate, and then neutralized with acetic acid. The aqueous layer was extracted with dichloromethane and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the dichloromethane layer was concentrated, the resulting crude powder was washed with normal hexane, and the insoluble matter was dried to give 1- (2-hydroxy-4,6-dimethoxyphenyl) -hexane-1, 0.71 g of 3-dione was obtained as a yellow powder. 650 mg of the obtained powder was suspended in 5 mL of concentrated hydrochloric acid, heated at 70 ° C. for 5 minutes, allowed to cool to room temperature, poured into 30 mL of ice water, and stirred for a while. The mixture was neutralized with 2N aqueous sodium hydroxide, extracted with dichloromethane, and the organic layer was washed with water and dried over anhydrous magnesium sulfate. The desiccant was filtered off and concentrated, and the resulting yellow crude powder was washed with normal hexane, and the insoluble matter was dried to obtain 550 mg of 5,7-dimethoxy-2-propylchromen-4-one as a yellow powder. It was.

(Step 9)
8.00 g of 3,5-dimethoxyphenol and 7.40 g of methyl 3-methylcrotonate were added to 60 mL of methanesulfonic acid preheated to 70 ° C. and stirred for 2.5 hours. The reaction solution was allowed to cool to room temperature, poured into ice water, stirred for a while, and extracted four times with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by column chromatography (Wakogel C-200, 150 g, developed with normal hexane: ethyl acetate = 1: 1) to give 5,7-dimethoxy-4,4-dimethyl-chroman-2-one as a yellow oil. As a result, 4.55 g was obtained.

(Process 10)
3.50 g of 3,5-dimethoxyphenol and 3.75 g of methyl 3-cyclopropyl-3-oxopropionate were added to methanesulfonic acid preheated to 70 ° C. and stirred for 2.5 hours. The reaction solution was allowed to cool to room temperature, poured into ice water, stirred for a while, and extracted three times with dichloromethane. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. The obtained crude crystals were washed with normal hexane to obtain 2.30 g of 5,7-dimethoxy-4- (3-hydroxypropyl) -2H-chromen-2-one as pale yellow crystals.

(Step 11)
394 mg of 5,7-dimethoxy-4- (3-hydroxypropyl) -2H-chromen-2-one was suspended in 8 mL of dichloromethane, 0.26 mL of tigloyl chloride and 1.25 mL of tin tetrachloride were added in this order under ice cooling. The solution was added dropwise and stirred at 5 ° C. for 15 hours. The reaction solution was poured into ice water, stirred for a while, extracted with ethyl acetate, the organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and saturated brine in that order, and dried over magnesium sulfate. The desiccant is removed by filtration, and the residue obtained by concentrating the organic layer is purified by column chromatography (20 g of Wakogel C-200, developed in normal hexane: ethyl acetate = 4: 1 to 1: 1), whereby the target compound is obtained. 0.25 g of 3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate (BNS-44) as a colorless powder Melting point: 87 ° C.) and 3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl which is the target compound -0.50 g (melting point: 88 ° C) of (2E) -2-methylbut-2-noate (BNS-45) was obtained as a colorless powder. Using a similar method, the target compound is
5,7-dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2-propyl-4H-chromen-4-one (BNS-46),
5,7-dimethoxy-4,4-dimethyl-8-[(2E) -2-methylbut-2-noyl] chroman-2-one (BNS-47), and 4- (4-chlorophenyl) -5,7 -Dimethoxy-8-[(2E) -2-methylbute-2-noyl] -2H-chromen-2-one (BNS-48) was obtained.

Example 2

(Process 1)
Preparation of 5,7-dimethoxy-4-propylquinolin-2 (1H) -one 1.00 g of 3,5-dimethoxyaniline and 1.03 g of ethyl butyryl acetate were dissolved in 20 mL of toluene, and 10 drops of mesylic acid were added. Heated to reflux. Ice water was added to the reaction solution, and the precipitated crystals were collected by filtration, washed with water and methanol, and then dried under reduced pressure. 1.05 g of the target product was obtained as white crystals.
(Process 2)
Preparation of 5,7-dihydroxy-4-propylquinolin-2 (1H) -one 0.5 g of 5,7-dimethoxy-4-propylquinolin-2 (1H) -one was dissolved in 10 mL of methylene chloride and cooled with ice. Then, 0.45 mL of boron tribromide was added dropwise. The resulting mixture was returned to room temperature for 1 hour at 0 ° C. and stirred for 30 minutes. The reaction solution was poured into ice water, and the precipitated crystals were collected by filtration, washed with water and dried. The crude crystals were washed with ethyl acetate to obtain 347 mg of the desired product.
(Process 3)
Preparation of 5,7-dihydroxy-8-propionyl-4-propylquinolin-2 (1H) -one 0.52 g of 5,7-dihydroxy-4-propylquinolin-2 (1H) -one and 1.28 g of aluminum chloride A suspension of 1,2-dichloroethane (20 mL) was heated and stirred at 60 ° C. for 30 minutes. A 1,2-dichloroethane solution (8 mL) of 0.37 g of propionic anhydride and 0.64 g of aluminum chloride was added dropwise to this suspension. The mixture was stirred with heating at 60 ° C. for 1 hour and at 75 ° C. for 2 hours. Ice water was added to the reaction solution, acidified with 10% hydrochloric acid, and left overnight. The precipitated crystals were collected by filtration, washed with water and dried. The obtained crude crystals were washed with methanol to obtain 266 g of the desired product.
(Process 4)
Preparation of 5-hydroxy-2,2-dimethyl-6-propionyl-10-propyl-2,7-dihydro-8H-pyrano [2,3-f] quinolin-8-one 5,7-dihydroxy-8-propionyl To 254 mg of -4-propylquinolin-2 (1H) -one and 413 mg of 4,4-dimethoxy-2-methyl-2-butanol, 0.5 mL of pyridine was added, and the mixture was heated and stirred at 160 ° C. overnight. Ice water was added to the reaction mixture, acidified with 10% hydrochloric acid, and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO ₄ ), and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 2/1 → 1/2). The obtained crude crystals were washed with ethyl acetate and dried to obtain the target compound, 5-hydroxy-2,2-dimethyl-6-propionyl-10-propyl-2,7-dihydro-2H, 8H-pyrano [2, 189 mg of 3-f] quinolin-8-one (BNS-10) was obtained as yellow crystals (melting point: 240-242 ° C.).

The definition of each symbol is as defined above.
(Process 5)
Dissolve 200 mg of 8-quinolinecarboxylic acid, 242 mg of 4-trifluoromethylaniline, and 441 mg of 2-chloro-N-methylpyridinium iodide in 3 mL of DMI (N, N′-dimethylimidazolidone), add 297 mg of ethyldiisopropylamine at room temperature. After stirring for 16 hours, water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and then the desiccant was filtered off and concentrated. The residue was purified by column chromatography (50 g of Wakogel C-200, developed in normal hexane: ethyl acetate = 4: 1 to 1: 1), and the resulting crude crystals were dissolved in ethyl acetate / petroleum ether and allowed to stand at room temperature for re-treatment. Crystallized. The precipitate was collected by filtration, washed with cold petroleum ether, and then dried under reduced pressure to obtain 202 mg of the target compound, N- [4- (trifluoromethyl) phenyl] quinoline-8-carboxamide (BNS-7). (Melting point: 114.7 ° C). In the same manner, 25 mg of 4- (ethylamino) -6-methyl-N- [4- (trifluoromethyl) phenyl] quinazoline-8-carboxamide (BNS-11), the target compound, was obtained (melting point 216 .5-217.5 ° C).

The definition of each symbol is as defined above.
(Step 6)
10 mL of phosphorus oxychloride was added to 1.90 g of 3,5-dimethoxyphenylacetic acid and 1.31 g of N-methylbenzamide, and the mixture was heated and stirred on a 110 ° C. oil bath for 5 hours. After cooling at room temperature, the reaction mixture was poured into ice water, stirred for a while, and extracted with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium hydrogen carbonate solution, dried over magnesium sulfate, and the desiccant was filtered off and concentrated. The residue was purified by column chromatography (80 g of Wakogel C-200, eluted with 0.1 to 1% methanol-containing dichloromethane), and the resulting crude powder was washed with normal hexane to obtain the target compound, 6,8-dimethoxy. 0.90 g of 2-methyl-1-phenylisoquinolin-3 (2H) -one (BNS-33) was obtained as a yellow powder (melting point: 195-197 ° C.).
Using the same method, 6,7,8-trimethoxy-2-methyl-1-phenylisoquinolin-3 (2H) -one which is the corresponding target compound from 4.50 g of 3,4,5-trimethoxyphenylacetic acid 0.78g of (BNS-34) was obtained as a yellow powder (melting point: 156-159 ° C).

  The physical properties of the target compounds are shown in Tables 1 to 9 below.

Test Example 1 Cell growth inhibitory action (1)
(Method)
The human leukemia cell line BV173 (obtained from ATCC) was used to evaluate the cell growth inhibitory action of the compound of the present invention. BV173 was maintained in an RPMI-1640 culture medium (GibcoBRL) supplemented with inactivated 10% FCS (Hyclone) at 37 ° C. in a humidified atmosphere of 5% CO ₂ .
Cell proliferation was determined using a modified MTT assay with cell counts using the trypan blue exclusion method and SF reagent (Nacalai Tesque).
BV173 was cultured as 2 × 10 ⁴ cells per well in a 100 μl medium in a flat bottom 96 well plate (Greiner laboratory) and cultured with various concentrations of compound for 72 hours. The modified MTT assay was applied to the cultured cells, and the cell growth inhibitory action of each compound was measured.
(result)
The results are shown in Table 10.

BNS-22 inhibited the growth of many cancer cell lines with IC ₅₀ = 1-5 μM.
Furthermore, about the BNS-22 by which the outstanding cancer cell growth inhibitory effect was recognized, the growth inhibitory effect was evaluated using the human cancer cell panel. As a human cancer cell panel, 39 types of cancer cells derived from various human organs were used [breast cancer (HBC-4, BSY-1, HBC-5, MCF-7, MDA-MB-23), central nervous cancer. (U251, SF-268, SF-295, SF-539, SNB-75, SNB-78), colon cancer (HCC2998, KM-12, HT-29, HCT-15, HCT-116), lung cancer (NCI- H23, NCI-H226, NCI-H522, NCI-H460, A549, DMS273, DMS114), melanoma (LOX-IMVI), ovarian cancer (OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, SK-OV) -3), renal cancer (RXF-631L, ACHN), gastric cancer (St-4, MKN1, MKN7, MKN28, MKN45, MKN7) ), Prostate cancer (DU-145, PC-3)]. All of them are collected as a human cultured cancer cell panel and are known and can be obtained commercially or from a cell bank or the like.
The cancer-killing cell activity possessed by BNS-22 can be confirmed by the following method. BNS-22 is added to all cell lines, and an in vitro drug sensitivity test is performed to test the cell growth inhibitory effect and the cell killing effect. For example, after adding BNS-22, the growth rate is measured colorimetrically by multiplying the number of living cells 48 hours later by staining with the protein-binding dye sulforhodamine B. When BNS-22 sensitivity measurement data of all human cultured cells are input to a computer, the concentration at which cell growth is inhibited by 50% (GI ₅₀ ), the concentration at which cell growth is apparently completely suppressed (TGI), and the number of cells are inoculated The concentration (LC ₅₀ ) is reduced to 50%, and the sensitivity to the drug is evaluated by performing information processing. The cancer-killing cell activity of the compound of the present invention can be examined using the above method. The original method is a known method described in ["Cancer and Chemotherapy" 24 (2): 129-135, 1997].
BNS-22 inhibited the proliferation of all 39 types of human cancer cells, and its inhibition profile was low in homology with existing anticancer agents, suggesting the possibility of having a novel mechanism of action. On the other hand, the inhibitory action of BNS-22 on normal fibroblast NHF was IC ₅₀ = 51.5 μM.

Test Example 2 Effect on Cell Cycle In HeLa cells, the effect of BNS-22 on the cell cycle was examined. HeLa cells (RIKEN Cell Bank) are DMEM (dulbeco's modified eagle's medium) containing 1% penicillin / streptomycin solution (manufactured by SIGMA), 10% fetal bovine serum (manufactured by JRH Biosciences), 1% sodium pyruvate (manufactured by Gibco); D5796 manufactured by SIGMA) and maintained in a humidity culture at 37 ° C. and 5% CO ₂ .
(Method)
(1) Immunostaining HeLa cells were treated with BNS-22 (10 μM) for 12 hours. BNS-22 was dissolved in DMSO and used. After the treatment, the cells were fixed with methanol and immunostained with DAPI (manufactured by SIGMA), anti-β tubulin antibody (manufactured by SIGMA), and anti-γ tubulin antibody (manufactured by SIGMA). DAPI indicates the presence of the nucleus. The anti-β tubulin antibody is an antibody against tubulin, a constituent protein of microtubules, and the anti-γ tubulin antibody is an antibody against γ tubulin, a constituent protein of the centrosome. The state of microtubules during mitosis can be known by immunostaining with these antibodies.
(2) FACS analysis HeLa cells were treated with BNS-22, and the cells were collected and rinsed with PBS, followed by DNA staining with 2 μg / mL prodium iodide (PI) for 30 minutes. After staining, analysis was performed by flow cytometry (FACScan; Becton Dickinson, NJ). The measurement was performed when the concentration of BNS-22 was changed for 24 hours (FIG. 2a) and when the treatment time was changed with BNS-22 (3 μM) (FIG. 2b).
(result)
BNS-22 was shown to cause mitotic abnormalities specifically in M phase cells in HeLa cells (FIG. 1).
BNS-22 was also shown to induce G2 / M phase arrest and polyploidy (FIG. 2).

Test Example 3 Cell growth inhibitory action (2)
About BNS-22, the cell growth inhibitory effect was investigated using the HeLa cell.
(Method)
Cell proliferation was measured using the WST-8 method. The WST-8 method is a method for measuring the number of living cells using the fact that tetrazolium salt WST-8 is reduced and yellow formazan is produced in the presence of an electron carrier (1-methoxy-PMS) in living cells. It is.
HeLa cells were cultured as 1 × 10 ⁴ cells per well in a 200 μl medium in flat bottom 96 well plates (Greiner laboratory) and cultured with various concentrations of BNS-22 for 24 hours.
The cell growth inhibitory action of BNS-22 was examined by applying the WST-8 method to the cultured cells.
(result)
The results are shown in FIG. A cell growth inhibitory effect of IC ₅₀ = 2 μM was observed by BNS-22.

Test Example 4 Topoisomerase II Activity Inhibitory Action Topoisomerase II is an enzyme that eliminates knots and knots of linked DNA that requires cleavage and passage of double-stranded DNA. As typical inhibitors, etoposide (VP-16) and amrubicin are known. The mechanism of action of topoisomerase inhibitors is to cleave DNA, then form a complex with topoisomerase II, inhibit DNA recombination, inhibit DNA replication, and inhibit G2 / M phase progression. Are known.
The influence of the compound of the present invention on topoisomerase II activity was examined. BNS-22 was used as the compound of the present invention.
(Method)
In vitro topoisomerase II inhibitory activity was measured using Topoisomerase II assay kit (TopoGEN, Inc.) and human topoisomerase II (TopoGEN Inc.). The reaction was performed at 37 ° C. for 30 minutes using a predetermined concentration of BNS-22. Other detailed procedures were performed according to the instructions attached to the kit.
(result)
The results are shown in FIG. Compared to etoposide, which is known as a representative inhibitor of topoisomerase II, stronger topoisomerase II inhibitory activity was observed in BNS-22.

Test Example 5 Search for binding protein using BNS-22 affinity beads (method)
Preparation of BNS-22 affinity beads NHS-activated sepharose ^™ 4 Fast Flow (manufactured by GE-Healthcare) was used. First, 200 μl of beads were measured in two 1 ml eppenes, and the beads were washed 5 times with 1 mM hydrochloric acid aqueous solution. Next, it was washed with a reaction buffer (0.1 M NaHCO ₃ aq-dioxane (1: 1 v / v). 11.1 mg of photoaffinity linker; N- (2- (2- (2- (2- 200 μl of reaction buffer 400 μl containing (2- (2- (2-aminoethoxy) ethoxy) ethoxy) ethoxy) ethoxy) ethyl) -4- (3- (trifluoromethyl) -3H-diazirin-3-yl) benzamide) Added to the beads, mixed with beads using a rotator for 2 hours at room temperature under light-shielded conditions, and introduced a photoaffinity linker into the beads. 1M ethanolamine solution was added to the mixture, and mixed with a rotator for 1 hour at room temperature under light-shielded conditions to block the beads.The beads were washed several times with MQ, suspended in MQ, and spin column (AM) Moved to Scan & Mightly Columns for DNA GEL Filtration After removing MQ by centrifugal operation, methanol was added to the spin column, and methanol was removed by centrifugal operation.This operation was repeated several times, and the beads were washed with methanol.10 ml sample bottle (manufactured by Nippon Denka Glass) One side was added with a methanol solution containing 1.3 mg of BNS-22, one side was added with methanol alone, and the dried beads were added and mixed with each. Propanol was added, azeotroped, and dried under reduced pressure for 0.5 hours.The dried beads were irradiated with UV at 365 nm 4 Jcm ^-2 using CL-1000L ultraviolet cross-linker (UVP-Inc., CA, USA). Transfer the beads to a spin column with methanol, MQ, and 50% methanol aqueous solution, and wash several times in the order of methanol, dimethyl sulfoxide, methanol, and PBS. Was added a PBS solution containing 1 mM NaN ₃ in 200 [mu] l, and stored at 4 ° C..
Preparation of cell extract A cell extract was prepared using HL-60 cells (RIKEN Cell Bank).
HL-60 cells were maintained in a RPMI1640 medium supplemented with 10% fetal calf serum in a 37 ° C., 5% CO ₂ humidified culture. The cultured HL-60 cells were collected and then precipitated by centrifugation, and the supernatant was removed. A binding buffer (10 mM Tris-HCl, 50 mM KCl, 5 mM MgCl ₂ , 1 mM EDTA, PH 7.6) to which a protease inhibitor (PI cocktail (Roches), PMSF (SIGMA)) was added was added, and a syringe with an injection needle (PH 7.6) was added. It was suspended using Terumo). After the suspension, the cells were disrupted with ultrasonic waves, and the insoluble fraction was precipitated by centrifugation (1500 rpm, 30 minutes, 4 ° C). The protein concentration of the obtained cell extract of the supernatant was determined using a protein assay kit (BIO RAD).
Search for coprecipitated protein The HL-60 cell extract prepared above was mixed with BNS-22 affinity beads or beads not immobilized with BNS-22 as a control, and specifically bound to BNS-22 affinity beads. Proteins were coprecipitated and identified. Proteins bound to the beads were solubilized, reduced, and separated by electrophoresis. The presence of the protein was examined by staining with Coomassie Brilliant Blue.
(result)
The results are shown in FIG.
The UBA-UBL type ubiquitin receptor ubiquitin 1 was identified as a protein that specifically binds to and co-precipitates with BNS-22 affinity beads.

Test Example 6 Accumulation of intracellular ubiquitinated proteins (1)
The effect of BNS-22 on intracellular ubiquitinated proteins was examined.
(Method)
HeLa cells were cultured in the presence of BNS-22 (3 μM) for 0, 3, 6, 9, 12, 18 and 24 hours, respectively. HL-60 cells were cultured in the presence of BNS-22 (1 μM) for 0, 12, 24 and 48 hours, respectively.
In addition, each cell was cultured for 2 hours in the presence of MG-132 (10 μM, Calbiochem), which is known to induce accumulation of ubiquitinated proteins (Trends Cell Biol., 8: 397-403, 1998).
A cell extract was prepared from the cultured cells and immunoblotted with an anti-ubiquitin antibody (Santa Cruz Biotechnology).
After recovering the cells treated with the drug, RIPA buffer (25 mM Hepes, 1.5% TX-100, 1.0% deoxycholic acid) containing protease inhibitors (PI cocktail (Roches), PMSF (SIGMA)) was added. Sodium, 0.1% SDS, 0.5 M NaCl, 5 mM EDTA, pH 7.8) was added, the cells were disrupted with ultrasound, and the insoluble fraction was precipitated by centrifugation (1400 rpm, 20 min). The protein concentration of the obtained cell extract of the supernatant was prepared using a protein assay kit (manufactured by BIO RAD), and the sample buffer (280 mM Tris-HCl, 44.8% glycerol, 12% SDS) of 1/4 of the total amount was prepared. , 20% 2-mercaptoethanol, 0.4 mg / ml bromophenol blue, pH 6.8) and heated at 100 ° C. for 3 minutes in a heat block was used as a sample. Using 7.5% polyacrylamide gel, proteins were separated by electrophoresis in electrophoresis buffer (25 mM Tris-HCl, 192 mM glycine, 0.1% SDS, pH 8.3), and the proteins contained in the gel were transferred. Transferred to a PVDF membrane (Millipore) in buffer (25 mM Tris-HCl, 192 mM glycine, 0.1% SDS, 20% methanol, pH 8.3). After the transfer, the PVDF membrane was immersed in TBS-T buffer (20 mM Tris-HCl, 137 mM NaCl, 0.1% Tween-20, pH 7.6) containing 5% skim milk, and blocked at room temperature for 0.5 hour. . Wash with TBS-T buffer for 1 hour, add 1/2000 total anti-ubiquitin antibody (manufactured by Santa Cruz Biotechnology) in TBS-T buffer containing 5% skim milk, react for 1 hour at room temperature, and again with TBS-T After washing with T buffer, a total amount of 1/2000 HRP-labeled anti-mouse antibody (Vector) was added to TBS-T buffer containing 5% skim milk, and reacted at room temperature for 1 hour. The target protein was detected by exposing a PVDF membrane washed with a TBS-T buffer for 1 hour at room temperature using a chemiluminescence reagent Supersignal west pico Chemiluminescence substrate (Pierce).
(result)
The results are shown in FIG. In any cell, BNS-22 was shown to induce the accumulation of ubiquitinated proteins.

Test Example 7 Search for binding protein using GUT-70 affinity beads (method)
GUT-70 affinity beads were produced in the same manner as in Test Example 5, except that GUT-70 was used as the compound to be bound to the beads.
The extract of HL-60 cells prepared in the same manner as in Test Example 5 was mixed with GUT-70 affinity beads or beads not immobilized with GUT-70 as a control, and specifically bound to GUT-70 affinity beads. Proteins were coprecipitated and identified. Proteins bound to the beads were solubilized, reduced, and separated by electrophoresis. The presence of the protein was examined by staining with Coomassie brilliant blue (pull-down assay).
(result)
A schematic diagram and results of the pull-down assay are shown in FIG.
Ubiquilin 1 was identified as a protein that specifically binds to and co-precipitates with GUT-70 affinity beads.

Test Example 8 Binding Affinity of GUT-70 to Ubiquitin 1 In Test Example 7, since ubiquitin 1 was identified as a protein specifically co-precipitated from the cell extract to GUT-70 affinity beads, GUT-70 The interaction between ubiquitin 1 and ubiquitin 1 was examined using purified recombinant ubiquitin 1.
Purified recombinant ubiquitin 1 was prepared as follows (His-ubiquitin 1).
The human ubiquitin 1 gene was cloned from a cDNA library of HL-60 cells and incorporated into a pRSET C vector (manufactured by Invitrogen). It was transformed into Escherichia coli BL21 (DE3) strain, and LB medium (5 g / l Yeast Extract, 5 g / l NaCl, 10 g / l Trypton) containing 1 mM isopropyryl-BD-thiogalactopyroside (IPTG), 2 ° C., 2 ° C. The expression of the His-ubiquitin 1 protein was induced by culturing for an hour, and purified recombinant His-ubiquitin 1 was prepared with AKTA purifier (manufactured by GE Healthcare). FIG. 8 (c) shows 1 μg of purified recombinant His-ubiquitin 1 subjected to SDS-PAGE and CBB staining.
A protein specifically bound to GUT-70 affinity beads was coprecipitated in the same manner as in Test Example 7 except that a His-ubiquitin 1-containing solution was used instead of the HL-60 cell extract. Similarly, when free GUT-70 (125 μg / ml) was added, the protein specifically bound to the GUT-70 affinity beads was coprecipitated. After solubilizing and reducing the protein bound to each bead, using 7.5% polyacrylamide gel, electrolysis in electrophoresis buffer (25 mM Tris-HCl, 192 mM glycine, 0.1% SDS, pH 8.3) Proteins were separated by electrophoresis, and proteins contained in the gel were transferred to a PVDF membrane (Milipore) in a transfer buffer (25 mM Tris-HCl, 192 mM glycine, 0.1% SDS, 20% methanol, pH 8.3). . After the transfer, the PVDF membrane was immersed in TBS-T buffer (20 mM Tris-HCl, 137 mM NaCl, 0.1% Tween-20, pH 7.6) containing 5% skim milk, and blocked at room temperature for 0.5 hour. . Wash with TBS-T buffer for 1 hour, add 1/5000 of anti-Xpress antibody (manufactured by Invitrogen) in TBS-T buffer containing 5% skim milk, react at room temperature for 1 hour, and again with TBS-T buffer. The total amount of 1/5000 HRP-labeled anti-mouse antibody (manufactured by Vector) was added to TBS-T buffer containing 5% skim milk and allowed to react at room temperature for 1 hour. The target protein was detected by exposing the PVDF membrane washed with TBS-T buffer at room temperature for 1 hour using a chemiluminescence reagent, Supersignal West Pico Chemiluminescence Substrate (manufactured by Pierce).
(result)
The results are shown in FIG. Since the purified recombinant ubiquitin 1 coprecipitates specifically on GUT-70 affinity beads and its binding is inhibited by the addition of free GUT-70 to the reaction system, GUT-70 directly binds to ubiquitin 1. It turns out that they are connected.

Test Example 9 Accumulation of intracellular ubiquitinated protein (2)
The effect of GUT-70 on intracellular ubiquitinated proteins was examined.
(Method)
HeLa cells were cultured in the presence of GUT-70 (50 μM) for 0, 0.5, 1, 3, 6, 12, 24 and 48 hours, respectively.
In addition, HeLa cells were cultured for 0, 0.5, 1, 3, 6, 12, 24, and 48 hours in the presence of MG-132 (1 μM, manufactured by Calbiochem), which is known to induce accumulation of ubiquitinated proteins.
Furthermore, as a control, HeLa cells were cultured with only solvent (DMSO) for 0, 0.5, 1, 3, 6, 12, 24 and 48 hours, respectively.
In the same manner as in Test Example 6, a cell extract was prepared from the cultured cells and immunoblotted with an anti-ubiquitin antibody.
(result)
The results are shown in FIG. GUT-70 has been shown to induce the accumulation of ubiquitinated proteins.
From the ubiquitin-binding properties shown in Test Examples 5, 7 and 8, and the accumulation-inducing action of ubiquitinated proteins shown in Test Examples 6 and 9, the compound of the present invention exhibits a ubiquitin-1 binding action and a ubiquitin-proteasome system inhibitory action. It was shown to have.

Although several specific embodiments of the present invention have been described in detail, those skilled in the art will recognize that various modifications may be made to the specific embodiments shown without departing from the teachings and advantages of the invention. It is possible to make changes. Accordingly, all such modifications and changes are intended to be included within the spirit and scope of the present invention as claimed.
This application is based on patent application Nos. 2007-287570 and 2008-120575 filed in Japan, the contents of which are incorporated in full in this application.

Claims (22)

A ubiquitin-binding small molecule, which is a compound represented by the following general formula (II):

{In formula (II)
R _{1 ′} is a heterocyclic group, or a C ₁ -C ₆ alkylamino group, a di (C ₁ -C ₆ ) alkylamino group, a C ₆ -C ₁₄ arylamino group, or a di (C ₆ -C ₁₄ ) arylamino. Group, an amino group having a substituent selected from the group consisting of phenylaminocarbonyl, butenylcarbonyl, and benzodioxolylcarbonyl, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or a general formula (1 ′) :

[In the formula (1 ′),
R _{16 ′} and R _{18 ′} are the same or different and are hydrogen, alkyl group or halogen, cyano group, nitro group, hydroxy group, aminocarbonyl group, acetyl group, carboxyl group, alkoxycarbonyl group, amino group, alkoxy group, alkyl group. R _{17 ′} represents a halogen, a cyano group, a nitro group, a hydroxy group, an aminocarbonyl group, an acetyl group, a carboxyl group, an alkoxycarbonyl, and an aryl group optionally having a substituent selected from the group consisting of Means an aryl group which may have a substituent selected from the group consisting of a group and an amino group]
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents OR _{4a ′} (wherein R _{4a ′} represents an alkyl group),
formula:


The group represented by
Formula (a ′):


Wherein R _{5a ′} and R _{5b ′} are the same or different and have a substituent selected from the group consisting of a hydrogen or carboxyl group, a dialkylaminocarbonyl group, an alkyloxycarbonyl group, and an alkenylcarbonyloxy group. Represents a good alkyl group). 8 - (3,4-dihydro-quinoline -1 (2H) - ylcarbonyl) -5,7-dimethoxy-4-propyl--2H- chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
7-methoxy-8-[(2E) -2-methylbut-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate,
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate ,
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate,
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one, or 3- [8- ( 3,4-dihydro -2H- quinolin-1-yl-carbonyl) -5,7-dimethoxy-2-oxo -2H- chromen-4-yl] propionic acid methyl, Yu Bikirin binding small molecules. The compound represented by the general formula (II) is
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate, or 3- [8- The ubiquitin-binding small molecule according to claim 1, which is methyl (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate. molecule. 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromene, wherein the compound represented by the general formula (II) has the following structural formula The ubiquitin-binding small molecule according to claim 1, which is 2-one.
  The ubiquitin-binding small molecule according to claim 1, which is used for prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases. Topoisomerase II inhibitor, which is a compound represented by the following general formula (II):


{In formula (II)
R _{1 ′} is a heterocyclic group, or a C ₁ -C ₆ alkylamino group, a di (C ₁ -C ₆ ) alkylamino group, a C ₆ -C ₁₄ arylamino group, or a di (C ₆ -C ₁₄ ) arylamino. Group, an amino group having a substituent selected from the group consisting of phenylaminocarbonyl, butenylcarbonyl, and benzodioxolylcarbonyl, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or a general formula (1 ′) :

[In the formula (1 ′),
R _{16 ′} and R _{18 ′} are the same or different and are hydrogen, alkyl group or halogen, cyano group, nitro group, hydroxy group, aminocarbonyl group, acetyl group, carboxyl group, alkoxycarbonyl group, amino group, alkoxy group, alkyl group. R _{17 ′} represents a halogen, a cyano group, a nitro group, a hydroxy group, an aminocarbonyl group, an acetyl group, a carboxyl group, an alkoxycarbonyl, and an aryl group optionally having a substituent selected from the group consisting of Means an aryl group which may have a substituent selected from the group consisting of a group and an amino group]
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents OR _{4a ′} (wherein R _{4a ′} represents an alkyl group),
formula:


The group represented by
Formula (a ′):


Wherein R _{5a ′} and R _{5b ′} are the same or different and have a substituent selected from the group consisting of a hydrogen or carboxyl group, a dialkylaminocarbonyl group, an alkyloxycarbonyl group, and an alkenylcarbonyloxy group. Represents a good alkyl group). 8 - (3,4-dihydro-quinoline -1 (2H) - ylcarbonyl) -5,7-dimethoxy-4-propyl--2H- chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
7-methoxy-8-[(2E) -2-methylbut-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate,
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate ,
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate,
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one, or 3- [8- ( 3,4-dihydro -2H- quinolin-1-yl-carbonyl) -5,7-dimethoxy-2-oxo -2H- chromen-4-yl] propionate, preparative topoisomerase II inhibitors. The compound represented by the general formula (II) is
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate, or 3- [8- The topoisomerase II inhibitor according to claim 6, which is methyl (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate. . 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromene, wherein the compound represented by the general formula (II) has the following structural formula The topoisomerase II inhibitor according to claim 6, which is 2-one.

  The topoisomerase II inhibitor according to claim 6, which is used for prevention or treatment of at least one disease selected from the group consisting of cancer, neurodegenerative diseases, autoimmune diseases and infectious diseases. An anticancer agent, which is a compound represented by the following general formula (II):

{In formula (II)
R _{1 ′} is a heterocyclic group, or a C ₁ -C ₆ alkylamino group, a di (C ₁ -C ₆ ) alkylamino group, a C ₆ -C ₁₄ arylamino group, or a di (C ₆ -C ₁₄ ) arylamino. Group, an amino group having a substituent selected from the group consisting of phenylaminocarbonyl, butenylcarbonyl, and benzodioxolylcarbonyl, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or a general formula (1 ′) :

[In the formula (1 ′),
R _{16 ′} and R _{18 ′} are the same or different and are hydrogen, alkyl group or halogen, cyano group, nitro group, hydroxy group, aminocarbonyl group, acetyl group, carboxyl group, alkoxycarbonyl group, amino group, alkoxy group, alkyl group. R _{17 ′} represents a halogen, a cyano group, a nitro group, a hydroxy group, an aminocarbonyl group, an acetyl group, a carboxyl group, an alkoxycarbonyl, and an aryl group optionally having a substituent selected from the group consisting of Means an aryl group which may have a substituent selected from the group consisting of a group and an amino group]
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents OR _{4a ′} (wherein R _{4a ′} represents an alkyl group),
formula:

The group represented by
Formula (a ′):


Wherein R _{5a ′} and R _{5b ′} are the same or different and have a substituent selected from the group consisting of a hydrogen or carboxyl group, a dialkylaminocarbonyl group, an alkyloxycarbonyl group, and an alkenylcarbonyloxy group. Represents a good alkyl group). 8 - (3,4-dihydro-quinoline -1 (2H) - ylcarbonyl) -5,7-dimethoxy-4-propyl--2H- chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
7-methoxy-8-[(2E) -2-methylbut-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate,
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate ,
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate,
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one, or 3- [8- ( 3,4-dihydro -2H- quinolin-1-yl-carbonyl) -5,7-dimethoxy-2-oxo -2H- chromen-4-yl] propionate, anticancer agents. The compound represented by the general formula (II) is
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate, or 3- [8- The anticancer agent according to claim 11, which is methyl (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate. 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromene, wherein the compound represented by the general formula (II) has the following structural formula The anticancer agent according to claim 11, which is 2-one.

  The anticancer agent according to any one of claims 11 to 14, which has binding activity with ubiquitin and exhibits anticancer activity.   Cancer is acute myeloid leukemia, acute lymphocytic leukemia, malignant lymphoma, choriocarcinoma, multiple myeloma, soft tissue tumor, small cell lung cancer, chronic myelogenous leukemia, medullary thyroid cancer, osteosarcoma, head and neck cancer, esophagus The cancer according to any one of claims 11 to 15, which is selected from the group consisting of cancer, non-small cell lung cancer, colorectal cancer, gastric cancer, biliary tract cancer, brain tumor, malignant melanoma, kidney cancer, pancreatic cancer and liver cancer. Anticancer drugs. The compound represented by the following general formula (II) or a pharmaceutically acceptable salt thereof:


{In formula (II)
R _{1 ′} is a heterocyclic group, or a C ₁ -C ₆ alkylamino group, a di (C ₁ -C ₆ ) alkylamino group, a C ₆ -C ₁₄ arylamino group, or a di (C ₆ -C ₁₄ ) arylamino. Group, an amino group having a substituent selected from the group consisting of phenylaminocarbonyl, butenylcarbonyl, and benzodioxolylcarbonyl, a heterocyclic carbonyl group, a cycloalkylcarbonyl group, a cycloalkenylcarbonyl group, or a general formula (1 ′) :

[In the formula (1 ′),
R _{16 ′} and R _{18 ′} are the same or different and are hydrogen, alkyl group or halogen, cyano group, nitro group, hydroxy group, aminocarbonyl group, acetyl group, carboxyl group, alkoxycarbonyl group, amino group, alkoxy group, alkyl group. R _{17 ′} represents a halogen, a cyano group, a nitro group, a hydroxy group, an aminocarbonyl group, an acetyl group, a carboxyl group, an alkoxycarbonyl, and an aryl group optionally having a substituent selected from the group consisting of Means an aryl group which may have a substituent selected from the group consisting of a group and an amino group]
Means a structure represented by
R _{2 ′} means an alkoxy group;
R _{3 ′} means hydrogen or an alkyl group;
R _{4 ′} represents OR _{4a ′} (wherein R _{4a ′} represents an alkyl group),
formula:

The group represented by
Formula (a ′):


Wherein R _{5a ′} and R _{5b ′} are the same or different and have a substituent selected from the group consisting of a hydrogen or carboxyl group, a dialkylaminocarbonyl group, an alkyloxycarbonyl group, and an alkenylcarbonyloxy group. Represents a good alkyl group). 8 - (3,4-dihydro-quinoline -1 (2H) - ylcarbonyl) -5,7-dimethoxy-4-propyl--2H- chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclohex-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
7-methoxy-8-[(2E) -2-methylbut-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1H-imidazol-4-yl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
3- (5,7-dimethoxy-2-oxo-2H-chromen-4-ylpropyl)-(2E) -2-methylbut-2-noate,
3- {5,7-dimethoxy-8-[(2E) -2-methylbut-2-noyl] -2-oxo-2H-chromen-4-yl} propyl- (2E) -2-methylbut-2-enoate ,
8- (2,3-dihydro-1H-indo-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,6-dimethoxybenzamide,
N- (5,7-dimethoxy-2-oxo-4-propyl-2H-chromen-8-yl) -2,2-diphenylacetamide,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate,
5,7-diisopropoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8-[(2E) -3- (2,6-dimethoxyphenyl) prop-2-noyl] -5,7-dimethoxy-4-propyl-2H-chromen-2-one, or 3- [8- ( 3,4-dihydro -2H- quinolin-1-yl-carbonyl) -5,7-dimethoxy-2-oxo -2H- chromen-4-yl] propionate, of compound or a pharmaceutically tolerated Salt. The compound represented by the general formula (II) is
8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
5,7-dimethoxy-8-[(2E) -3-phenylprop-2-noyl] -4-propyl-2H-chromen-2-one,
8- (1-cyclopent-1-ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromen-2-one,
Ethyl 3- [8- (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] propionate, or 3- [8- The compound according to claim 17, which is (3,4-dihydro-2H-quinolin-1-ylcarbonyl) -5,7-dimethoxy-2-oxo-2H-chromen-4-yl] methyl propionate, or a pharmacy thereof Acceptable salt. 8- (3,4-dihydroquinolin-1 (2H) -ylcarbonyl) -5,7-dimethoxy-4-propyl-2H-chromene, wherein the compound represented by the general formula (II) has the following structural formula The compound or pharmaceutically acceptable salt thereof according to claim 17, which is 2-one.

  The pharmaceutical which contains the compound or its pharmaceutically acceptable salt of any one of Claims 17-20 as an active ingredient. The medicament according to claim 21, which is an anticancer agent.