JPH1045594A - Scavenger for active oxygen - Google Patents

Abstract

(57) [Summary] (with correction) [PROBLEMS] To provide a preventive and therapeutic agent for various diseases caused by active oxygen. SOLUTION: The following formula (I) Wherein R is hydrogen, carbamoyl, phenyl, 4-
Methylphenyl group, alkyl group having 1 to 4 carbon atoms or hydroxyl group, carboxyl group, carbamoyl group, methylmercapto group, phenyl group, 4-hydroxyphenyl group, mercapto group, imidazolyl group, amino group, guanidino group, methoxy group or phosphorus 1 carbon atom substituted with an acid group
Alkyl groups, R ¹ , R ² , and R ³ represent hydrogen, an alkyl group, an allyl group, a propargyl group, a benzyl group, a phenyl group, and an alkoxycarbonylmethyl group]. Active oxygen scavenger contained.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an active oxygen scavenger containing an imidazopyrroloquinoline compound as an active ingredient.

[0002]

2. Description of the Related Art Aerobic organisms, including humans, inhale oxygen and oxidize biological substances.
Used for ATP synthesis. In those processes, some of the oxygen is activated, producing so-called "active oxygen". Active oxygen includes O _2- (superoxide), H ₂ O ₂ (hydrogen peroxide), .OH (hydroxyl radical), 1 O ₂ (singlet oxygen), lipid peroxy radical (ROOH, derivatives of H ₂ O ₂ ) ), Lipid alkoxy radicals (RO, .OH derivatives), and the like. It has been clarified that these active oxygens peroxidize lipids in the living body, and the generated lipid peroxide causes various obstacles to the living body. The disorders include carcinogenesis, arteriosclerosis, diabetes, ischemia-reperfusion injury, Parkinson's disease, shock, disseminated intravascular coagulation, oral disease, lung disease, gastrointestinal disease, liver disease, kidney disease, cataract and bipyridyl Herbicides, halogenated hydrocarbons, benzine, chloroform and other drug addictions, aging, adult diseases and metabolic diseases are widespread. Alloxan, streptozotocin, adriamycin, daunomycin, anticancer drugs, methyldopa, a hypotensive drug,
Hydrazine, primaquine and pamaquine as antimalarial agents, paracetamol and phenacetin as analgesics, and hydrocortisone as anti-inflammatory and anti-allergic agents can cause diabetes, hepatic metabolism disorder, hemolysis, kidney disorder or cataract as side effects. However, these are considered to be disorders caused by active oxygen caused by drug administration.

On the other hand, the living body has a defense mechanism against harmful active enzymes, and a group of antioxidant enzymes such as glutathione peroxidase, catalase and superoxide dismutase and non-enzyme small molecules such as vitamin E, ascorbic acid and glutathione. There are a group of antioxidants that are thought to function in a complex relationship with each other.

In addition, it has been revealed that pyrroloquinoline quinone (PQQ), which has recently been discovered as a new coenzyme for oxidoreductase, has a strong active oxygen scavenging effect (Japanese Pharmaceutical Society 109th Annual Meeting, Abstracts V, Page 42 (198
9)) Furthermore, it has been reported to suppress liver damage induced by carbon tetrachloride, endotoxin shock, and hydrocortisone-induced cataract, which are considered to be caused by active oxygen. (Watanabe et al., Current Therapeutic Re
search, Vol. 44, pp. 896-901, 1988,
Matsumoto et al., "Jongejan, Duine, PQQ and Quinoprot
ein ", pp. 162-164, 1989 (Kluwer Acade
mic Publushers), Nishigori et al., Life Science, No. 4.
5, 593-598, 1989), and in recent years, the physiological action of PQQ in living organisms has been vigorously elucidated.

[0005] Since PQQ has a strong active oxygen scavenging effect, it is expected to be used as a therapeutic agent for various diseases considered as active oxygen disorders, but PQQ has been shown to have nephrotoxicity. (Watanabe et al., Hiro
Shima J. Med. Sci., Vol. 38, No. 1, pp. 49-51 (1987)). Therefore, toxicity and nephrotoxicity are low,
If a drug having an active oxygen scavenging action can be found, it is expected that it can be developed as a preventive and therapeutic drug for various aging, adult diseases and metabolic diseases, which are disorders caused by active oxygen.

[0006]

Means for Solving the Problems The present inventors have conducted intensive studies on drugs having a low toxicity and an active oxygen scavenging action for the above-mentioned reasons. As a result, imidazopyrroloquinolines or their esters were found It has been found that it has low toxicity, has a strong active oxygen scavenging effect, and has a preventive and therapeutic effect on various diseases that are active oxygen disorders. Imidazopyrroloquinoline (hereinafter referred to as IPQ) is 7,10-dihydro-7-oxo-imidazo [4,
5,1-ij] -pyrrolo [2,3-f] quinoline-1,
3,9-tricarboxylic acid, which is used in the present invention.
Substituted IPQs and their esters are represented by the following general formula (I).

[0007]

Embedded image [Where R is hydrogen, carbamoyl group, phenyl group, 4
A methylphenyl group, an alkyl group having 1 to 4 carbon atoms or a hydroxyl group, a carboxyl group, a carbamoyl group, a methylmercapto group, a phenyl group, a 4-hydroxyphenyl group,
1 carbon atom substituted by a mercapto group, imidazolyl group, amino group, guanidino group, methoxy group or phosphate group
And 4 to 4 alkyl groups. Provided that R ¹ , R ² and R ³ represent hydrogen, an alkyl group, an allyl group, a propargyl group, a benzyl group, a phenyl group and an alkoxycarbonylmethyl group.

[0008]

DETAILED DESCRIPTION OF THE INVENTION IPQ used in the present invention
And the IPQ ester compound can be produced by the following method. IPQs can be produced by reacting PQQs or salts thereof (hereinafter, referred to as PQQs) with various α-amino acids, methylamines and the like in the presence of oxygen. Also, certain IPQs have the formula (II)

[0009]

Embedded image Pyrroloquinoline (M. Mure, Tetrahedron Let
t., 30 , 6875-6878, 1989) and various aldehydes (R
-CHO).

The IPQ compound wherein R is H is a compound of 7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid ( It is synthesized from PQQ and at least one of glycine, tryptophan, proline, threonine, tyrosine, serine and monomethylamine, and from PQQ and ammonia and formaldehyde. The IPQ compound wherein R is CH ₃ is 7,10-dihydro-5-methyl-7-oxo-imidazo [4,
5,1-ij] pyrrolo [2,3-f] quinoline-1,
3,9-Tricarboxylic acid (hereinafter referred to as methyl IPQ), which is synthesized from PQQ and alanine. R is CH ₂ OH
Is 7,10-dihydro-5-hydroxymethyl-7-oxo-imidazo [4,5,1-ij]
Pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as hydroxymethyl IPQ)
It is synthesized from QQ and serine. R is I of CH (CH ₃ ) ₂
The PQ compound is 7,10-dihydro-5- (1-methylethyl) -7-oxo-imidazo [4,5,1-ij].
Pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as methylethyl IPQ) and PQQ
And valine.

An IPQ compound in which R is CH ₂ CH (CH ₃ ) ₂ is 7,10-dihydro-5- (2-methylpropyl)
-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 2-methylpropyl IPQ) and PQQ
And leucine. R is CH (CH ₃ ) CH ₂ C
IPQ compounds of H ₃ is 7,10-dihydro-5- (1
-Methylpropyl) -7-oxo-imidazo [4,5
1-ij] pyrrolo [2,3-f] quinoline-1,3,9
Tricarboxylic acid (hereinafter referred to as 1-methylpropyl IPQ), which is synthesized from PQQ and isoleucine. The IPQ compound in which R is CH ₂ CH ₂ CO ₂ H is 5- (2-carboxyethyl) -7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f Quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 2-carboxyethyl IPQ), which is synthesized from PQQ and glutamic acid. The IPQ compound in which R is CH ₂ CH ₂ CONH ₂ is 5- (2-carbamoylethyl) -7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f]. Quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 2-carbamoylethyl IPQ), which is synthesized from PQQ and glutamine.

The IPQ compound in which R is CH ₂ CH ₂ SCH _{3 is} a compound of 7,10-dihydro-5- (2-methylthioethyl) -7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3 -F] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 2-methylthioethyl IPQ)
It is synthesized from QQ and methionine. R is CH ₂ -C ₆ H ₅
Is a 5-benzyl-7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter benzyl) IPQ), which is synthesized with PQQ and phenylalanine. R is I of CH ₂ —C ₆ H ₄ —OH
The PQ compound is 7,10-dihydro-5- (4-hydroxyphenylmethyl) -7-oxo-imidazo [4,
5,1-ij] pyrrolo [2,3-f] quinoline-1,
3,9-tricarboxylic acid (hereinafter referred to as 4-hydroxyphenylmethyl IPQ), which is synthesized from PQQ and tyrosine. The IPQ compound in which R is CH ₂ SH is 7,10-
Dihydro-5-mercaptomethyl-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as methylcaptomethyl IPQ). Synthesized from PQQ and cysteine. The IPQ compound in which R is CH ₂ CO ₂ H is 5-carboxymethyl-7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3. , 9-tricarboxylic acid (hereinafter referred to as carboxymethyl IPQ), which is synthesized from PQQ and aspartic acid. R is

[0013]

Embedded image Is 7,10-dihydro-5- (4-imidazolylmethyl) -7-oxo-imidazo [4,5
1-ij] pyrrolo [2,3-f] quinoline-1,3,9
-Tricarboxylic acid (hereinafter referred to as 4-imidazolylmethyl IPQ)
Which is synthesized from PQQ and histidine. R
However, the IPQ compound of (CH ₂ ) ₄ NH ₂ is 5- (4-aminobutyl) -7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f]. Quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 4-aminobutyl IPQ), which is synthesized from PQQ and lysine.
R is

[0014]

Embedded image Is 5- (3-guanidinopropyl)-
7,10-dihydro-7-oxo-imidazo [4,5
1-ij] pyrrolo [2,3-f] quinoline-1,3,9
-Tricarboxylic acid (hereinafter referred to as 3-guanidinopropyl IPQ)
And synthesized from PQQ and arginine. R
Is an IPQ compound of CH ₂ CONH ₂ , which is 5- (carbamoylmethyl) -7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3 , 9-tricarboxylic acid (hereinafter referred to as carbamoylmethyl IPQ), which is synthesized from PQQ and asparagine. The IPQ compound in which R is CH ₂ OCH ₃ is 7,10
-Dihydro-5-methoxymethyl-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter methoxymethyl I
PQ) and synthesized from PQQ and O-methylserine. The IPQ compound in which R is CH ₂ OPO ₃ H ₂ is 7,
10-dihydro-7-oxo-5-phosphoryloxymethyl-imidazo [4,5,1-ij] pyrrolo [2,3-
f] Quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as phosphoryloxymethyl IPQ), which is synthesized from PQQ and O-phosphorylserine. R is the IP of CONH ₂
The Q compound is 5-carbamoyl-7,10-dihydro-
7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as carbamoyl IPQ), which is synthesized from PQQ and serinamide.

The IPQ compound in which R is CH ₂ CH ₃ is 5-ethyl-7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline-
1,3,9-Tricarboxylic acid (hereinafter referred to as ethyl IPQ), which is synthesized from a pyrroloquinoline compound represented by the formula (II) and propionaldehyde. R is CH ₂ C
The IPQ compound of H ₂ CH ₃ is 7,10-dihydro-7-
Oxo-5-propyl-imidazo [4,5,1-ij]
Pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as propyl IPQ), which is synthesized from a pyrroloquinoline compound represented by the formula (II) and butyraldehyde. The IPQ compound in which R is C ₆ H ₅ is 7,10-
Dihydro-7-oxo-5-phenyl-imidazo [4,
5,1-ij] pyrrolo [2,3-f] quinoline-1,
3,9-tricarboxylic acid (hereinafter referred to as phenyl IPQ)
And is synthesized from a pyrroloquinoline compound represented by the formula (II) and benzaldehyde. R is C ₆ H ₄ —CH ₃ I
The PQ compound is 7,10-dihydro-5- (4-methylphenyl) -7-oxo-imidazo [4,5,1-i
j] pyrrolo [2,3-f] quinoline-1,3,9-tricarboxylic acid (hereinafter referred to as 4-methylphenyl), which is synthesized from a pyrroloquinoline compound represented by the formula (II) and P-tolualdehyde . Incidentally, alkali metal salts, alkaline earth metal salts or hydrogen- or alkyl-substituted ammonium salts of these IPQ compounds are also included in the present invention (these compounds may be collectively referred to as IPQs hereinafter). Representative examples of the salt of the IPQ compound of the present invention include:
Alkali metal salts such as sodium salt or potassium salt, alkaline earth metal salts such as magnesium salt or calcium salt and ammonium salt, ammonium salt such as trimethylammonium salt, triethylammonium salt or triethanolammonium salt or alkyl-substituted ammonium salt. is there.

Physical properties of methyl IPQ 1) Elemental analysis: C ₁₆ H ₉ N ₃ O ₇ H ₂ O (MW 373.28) Theoretical value (%): C51.48 H2.97 N11.26 Actual value (%): C51 .20 H3.24 N11.03 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2725 ^{br, s} , 2480 ^{br, s} , 1590 ^s , 1515 ^s , 1170 ^vs , 1095
^{sh, s} , 965 ^m , 750 ^m , 710 ^m 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 2.73 (s, 3 H , CH ₃ ), 7.18 ( d, 1H, pyrrole ring C-
H , J = 1.8 Hz), 7.92 (s, 1H, pyridine ring C— H ), 12.82
(Brs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 255,272,418

Physical properties of hydroxymethyl IPQ 1) Elemental analysis: C ₁₆ H ₉ N ₃ O ₈ (MW 371.25) Theoretical value (%): C51.76 H2.44 N11.32 Actual value (%): C50.58 H2.63 N11.08 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2500 ^{br, s} , 1680 ^{sh, s} , 1575 ^{sh, s} , 1510 ^s , 1150 ^vs , 1095
^{sh, s} , 700 ^m , 650 ^m 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 4.98 (s, 2H, CH ₂ —OH), 7.27 (d , 1H, pyrrole ring C- H , J = 1.98 Hz), 8.07 (s, 1H, pyridine ring C- H ), 14.
60 (d, 1H, pyrrole ring NH , J = 0.44 Hz) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 254, 269 ^sh , 416

Physical properties of 1-methylethyl IPQ 1) Elemental analysis value: C ₁₈ H ₁₃ N ₃ O ₇ H ₂ O (MW 401.33) Theoretical value (%): C53.87 H3.77 N 10.47 Actual measurement value (%) ): C53.61 H3.96 N10.22 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2850 ^{br, s} , 2530 ^{sh, s} , 1575 ^s , 1515 ^{sh, m} , 1175 ^vs , 1100
^{sh, m} , 1015 ^{sh, m} , 815 ^{sh, w} , 755 ^m , 730 ^m , 660
^{sh, w} 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 1.36 (d, 6H, CH- (CH ₃ ) ₂ , J = 6.4 Hz), 3.74 (hep, 1H,
_{C H - (CH 3) 2} , J = 6.7Hz), 7.29 (d, 1H, pyrrole ring C
−H , J = 1.8 Hz), 8.00 (s, 1H, pyridine ring C− H ), 13.01 (b
rs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 255, 272 ^sh , 418

Physical properties of 2-methylpropyl IPQ 1) Elemental analysis: C ₁₉ H ₁₅ N ₃ O ₇ H ₂ O (MW 415.36) Theoretical value (%): C54.94 H4.13 N10.12 Actual value (%) ): C54.70 H4.27 N 9.85 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2880 ^{br, s} , 2520 ^{sh, s} , 2330 ^{sh, m} , 1585 ^s , 1520 ^{sh, m} ,
1180 ^vs , 760 ^{sh, m} , 735 ^m , 670 ^{sh, w} 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 0.93 (d, 6 H, CH ₂ -CH − (CH ₃ ) ₂ , J = 6.6 Hz), 2.16
_{(m, 1H, CH 2 -C} H - (CH 3) 2), 3.08 (d, 2H, C H 2
—CH— (CH ₃ ) ₂ , J = 7.0 Hz), 7.28 (d, 1 H, pyrrole ring C— H , J = 2.2 Hz), 7.99 (s, 1 H, pyridine ring C— H ), 12.60
(brs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 255, 272 ^sh , 419

Physical properties of 1-methylpropyl IPQ 1) Elemental analysis: C ₁₉ H ₁₅ N ₃ O ₇ H ₂ O (MW 415.36) Theoretical value (%): C54.94 H4.13 N10.12 Actual value (%) ): C54.68 H4.20 N 9.88 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2900 ^{br, s} , 2530 ^{sh, s} , 1605 ^s , 1520 ^{sh, m} , 1180 ^vs , 1145
^{sh, m} , 1100 ^{sh, m} , 850 ^w , 760 ^m , 725 ^{sh, m} , 670 ^{sh, w} 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 0.83 (t, 3H, CH (CH 3) -CH 2 -C H 3, J = 7.3H
z), 1.38 (d, 3H , CH (C H 3) -CH 2 -CH 3, J = 6.8
Hz), 1.71 (m, 2H , CH (CH 3) -C H 2 -CH 3), 3.54
(hex, 1H, C H ( CH 3) -CH 2 -CH 3, J = 6.5Hz), 7.
31 (d, 1H, pyrrole ring C- H, J = 2.4Hz), 7.98 (s, 1H, pyridine ring C- H), 13.07 (brs, 1H, pyrrole ring N- H) 4) Visible and Ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 256, 272 ^sh , 418

Physical properties of 2-carboxyethyl IPQ 1) Elemental analysis: C ₁₈ H ₁₁ N ₃ O ₉ H ₂ O (MW 431.31) Theoretical value (%): C50.12 H3.04 N 9.74 Actual value (%) : C49.85 H3.33 N 9.62 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2840 ^{br, s} , 2500 ^m , 2340 ^{sh, m} , 1570 ^s , 1515 ^s , 1175 ^vs ,
1100 ^{sh, m} , 755 ^w , 730 ^w 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 2.85 (t, 2H, CH ₂ —CH ₂ —CO ₂ —CO ₂ H, J = 6.2Hz), 3.30
_{(t, 2H, C H 2} -CH 2 -CO 2 H, J = 5.9Hz), 7.23 (brs,
1H, pyrrole ring C- H), 7.98 (s, 1H, pyridine ring C- H), 1
3.67 (brs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 256, 275 ^sh , 419

Physical properties of 2-carbamoylethyl IPQ 1) Elemental analysis: C ₁₈ H ₁₂ N ₄ O ₈ H ₂ O (MW 430.33) Theoretical value (%): C50.24 H3.28 N13.02 Actual measurement value (% ): C50.01 H3.50 N12.87 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2735 ^{br, m} , 2500 ^{br, m} , 2300 ^{sh, m} , 1555 ^{br, s} , 1390 ^s , 118
0 ^vs , 1100 ^{sh, s} , 990 ^m , 765 ^m , 745 ^m , 560 ^w 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 2.76 (t, 2H, _{CH 2 -C H 2 -CO-NH} 2, J = 6.7Hz), 3.
31 (t, 2H, C H 2 -CH 2 -CO-NH 2, J = 6.9Hz), 6.79
_{(brs, 1H, CH 2 -CH} 2 -CO-N H 2), 7.25 (d, 1H,
Pyrrole ring CH, J = 2.0Hz), 7.38 (brs, 1H, CH 2 -C
H ₂ —CO—N H ₂ ), 7.97 (s, 1 H, pyridine ring C— H ), 1
2.91 (brs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 255, 272 ^sh , 418

Physical Properties of 2-Methylthioethyl IPQ 1) Elemental Analysis: C ₁₈ H ₁₃ N ₃ O ₇ S.H ₂ O (MW 433.3
9) Theoretical value (%): C49.88 H3.49 N 9.70 Actual value (%): C49.63 H3.77 N 9.49 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 2775 ^{br, s} , 2500 ^{br, m} , 1575 ^s , 1510 ^{sh, s} , 1175 ^vs , 970
^w , 850 ^w , 750 ^m , 720 ^m 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 2.13 (s, 3 H, CH ₂ —CH ₂ —SC—C H ₃ ), 2.98 (t, 2H,
_{CH 2 -C H 2 -S-CH} 3, J = 7.7Hz), 3.45 (t, 2H, C
_{H 2 -CH 2 -S-CH 3} , J = 7.9Hz), 7.23 (d, 1H, pyrrole ring CH, J = 2.0Hz), 7.99 (s, 1H, pyridine ring CH), 13.
14 (brs, 1H, pyrrole ring NH ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 256, 273 ^sh , 418

Physical properties of benzyl IPQ 1) Elemental analysis: C ₂₂ H ₁₃ N ₃ O ₇ H ₂ O (MW 449.37) Theoretical value (%): C58.80 H3.36 N 9.35 Actual value (%): C58. 52 H3.61 N 9.14 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2710 ^{br, s} , 2490 ^{br, s} , 1575 ^{br, s} , 1490 ^{sh, s} , 1170 ^vs , 980
^w , 710 ^m , 690 ^m , 660 ^{sh, w} 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 4.67 (s, 2H, CH ₂ -C ₆ H) _{5), 7.02~7.90 (m, 6H} , pyrrole ring _{CH, CH 2 -C 6 H 5} ), 7.96 (s, 1H, pyridine ring CH), 12.95 (brs, 1H , pyrrole ring N-H 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 256, 273 ^sh , 419

[0025] 4-Hydroxyphenylmethyl IPQ
Property 1) Elemental analysis: C ₂₂ H ₁₃ N ₃ O ₈ H ₂ O (MW 465.37) Theoretical value (%): C56.78 H3.25 N 9.03 Actual value (%): C56.51 H3.52 N 8.87 2) IR spectrum (ν _max value, cm ^-1 ): (KBr) 2810 ^s , 2520 ^{s, sh} , 1590 ^s , 1500 ^s , 1415 ^m , 1180 ^vs , 9
90 ^{w, sh} , 820 ^{w, sh} , 760 ^m , 730 ^m , 670 ^{w, sh} 3) ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 4.54 (s, 2H) _{, C H 2 -C 6 H 4} OH), 6.64 (d, 2H, J = 8.6Hz,
CH ₂ side benzene ring C- H ) 6.81 (d, 2H, J = 8.1 Hz, OH side benzene ring C- H ) 7.31 (d, 1 H, pyrrole ring C- H , J = 2.0 Hz), 7.94 (s, 1H, pyridine ring C— H ), 13.01 (brs, 1H, pyrrole ring N— H ) 4) Visible / ultraviolet spectrum (λ _max value, nm): (10 mM potassium phosphate buffer pH 7.0) 256, 270 ^sh , 420

Physical properties of methoxymethyl IPQ ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 3.49 (s, 3H), 4.93 (s, 2H), 7.29 (s, 1H) ), 8.09 (s, 1
H) .Physical properties of phosphoryloxymethyl IPQ ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 5.47 (d, J = 9.2 Hz, 2H), 7.27 (s, 1H) ), 8.17 (s, 1H) .Physical properties of carbamoyl IPQ ¹ H-NMR spectrum (δ value, ppm): (DMSO-d ₆ , internal standard: TMS) 7.27 (s, 3H), 7.84 (s, 2H), 8.15 (s, 1H), 8.55 (s, 1
H).

The esters of IPQs are the same as those mentioned above.
In general formula (I), R¹, R^Two, R ^ThreeBut the same but different
May be a hydrogen atom, an alkyl group or an alkenyl group
Are benzyl groups, and are monoesters, diesters,
Or triester. Esthetics of these IPQs
In the normal form, IPQs or their salts and alcohols are
It can be easily manufactured by reacting more.
And alkyl groups such as methyl and ethyl
However, examples of the alkenyl group include an allyl group.
In addition, PQQ or PQQ salt and alcohols are
To give an ester of PQQs.
Reacting with certain amino acids or methylamine
It is also possible to obtain the desired IPQ esters
It is. Further, the esters of IPQs are represented by the general formula
In (I), R¹, R^Two, R^ThreeIs hydrogen, alkyl, a
Ril, propargyl, benzyl, phenyl and alkoxy
Indicates a carbonyl group, all of which are hydrogen
Except for IPQs-1,3,9-triester,
IPQs-1,3-diester (R^Three = H), IPQs
-1,9-diester (R^Two= H), IPQs-3,9
-Diester (R¹= H), IPQs-1-ester
(R^Two= R^Three= H), IPQs-3-esters (R1 =
R3 = H), IPQs-9-ester (R¹= R^Two= H)
But the types of the esters are the same or different.
May be. Esterification of IPQs according to the present invention
The following compounds can be mentioned as compounds.

Compound 1: IPQ-1,3,9-trimethylene
Ester (IPQ-TME) 7,10-dihydro-1,3,9-trimethoxycarbonyl-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline Compound 2: IPQ- 3,9-dimethyl ester (IPQ
-3,9-DME) 1-carboxy-7,10-dihydro-3,9-dimethoxycarbonyl-7-oxo-imidazo [4,5,1-
ij] pyrrolo [2,3-f] quinoline Compound 3: IPQ-1,9-dimethyl ester (IPQ
-1,9-DME) 3-carboxy-7,10-dihydro-1,9-dimethoxycarbonyl-7-oxo-imidazo [4,5,1-
ij] Pyrrolo [2,3-f] quinoline Compound 4: IPQ-9-methyl ester (IPQ-9-
ME) 1,3-Dicarboxy-7,10-dihydro-9-methoxycarbonyl-7-oxo-imidazo [4,5,1-
ij] pyrrolo [2,3-f] quinoline Compound 5: IPQ-3-methyl ester (IPQ-3-
ME) 1,9-Dicarboxy-7,10-dihydro-3-methoxycarbonyl-7-oxo-imidazo [4,5,1-
ij] pyrrolo [2,3-f] quinoline Compound 6: IPQ-1,3,9-triallyl ester
(IPQ-TAE) 1,3,9-triallyloxycarbonyl-7,10-dihydro-7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f] quinoline Compound 7: IPQ- 9-allyl ester (IPQ-9-
AE) 9-Allyloxycarbonyl-1,3-dicarboxy-
7,10-dihydro-7-oxo-imidazo [4,5
1-ij] pyrrolo [2,3-f] quinoline Compound 8: IPQ-1,3,9-tristearyl ester
(IPQ-TSE) 7,10-dihydro - 1,3,9-trioctadecyloxycarbonyl-7-oxo-imidazo [4,5,1-i
j] Pyrrolo [2,3-f] quinoline Compound 9: IPQ-9-stearyl ester (IPQ-
9-SE) 1,3-dicarboxy-7,10-dihydro-9-octadecyloxycarbonyl-7-oxo-imidazo [4,
5,1-ij] Pyrrolo [2,3-f] quinoline Compound 10: IPQ-1,3-diisopropyl ester
(IPQ-DIPE) 9-carboxy-7,10-dihydro-1,3-bis (1-methylethyloxycarbonyl) -7-oxo-imidazo [4,5,1-ij] pyrrolo [2,3-f ] Quinoline

The method for producing the above compound is described in detail below. The IPQ esters of compounds 1 to 9 correspond to the corresponding P
The ester compound of QQ is glycine or formalin /
It can be synthesized by treating with ammonium chloride. The synthesis of these PQQ esters can be obtained by selective esterification or hydrolysis from PQQ or a triester of PQQ, and furthermore, an appropriate combination thereof. For the esterification, a reaction using an acid catalyst using an alcohol, a reaction using an alkyl halide and a base, and further using a reaction aid such as an alcohol or an alkoxide and a 2-halobiridinium salt, dicarbonylimidazole, and dicyclohexylcarbodiimide. It can be performed by a reaction or the like. For the hydrolysis reaction, a reaction reagent such as trimethylsilyl iodide, lithium iodide, aluminum / alkylthiol halide, boron bromide, potassium cyanide and the like can be used in addition to the reaction under a base or acidic condition. Compounds 1, 6, 8
Can be obtained from IPQ in the same manner as PQQ triester, and compounds 4, 7, and
IPQ esters such as 9 can be synthesized from IPQ triesters by their selective hydrolysis. Compound 10 can be synthesized by selectively hydrolyzing IPQ monomethyldiisopropyl ester derived from compound 4. The IPQ ester compound can be purified by appropriately performing operations such as extraction, recrystallization, silica gel chromatography, reverse phase chromatography, gel filtration, concentration, centrifugation, and drying. IP
Means such as elemental analysis, nuclear magnetic resonance spectrum, infrared absorption spectrum, and ultraviolet / visible absorption spectrum are used to identify the Q-type ester compound. The quantitative determination of the IPQ ester compound can be performed by high performance liquid chromatography. The physical properties of the IPQ ester compound are shown below.

Physical properties of IPQ-TME 1) Melting point: 250 ° C. or more (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3450 ^{br, m} , 3178 ^w , 2960 ^w , 1731 ^s , 1662 ^vs , 1512
^m , 1259 ^vs , 1234 ^s , 991 ^m 3) ¹ H-NMR spectrum (δ value, ppm): 3.99 (s, 3H),
4.14 (s, 3H), 4.17 (s, 3H), 7.64 (d, J = 2.2Hz, 1H), 8.49
(s, 1H), 9.37 (s, 1H), 12.50 (br, 1H) 4) Visible / UV spectrum (λ _max value, nm): (in acetonitrile) 258, 275, 413 ^sh , 430 (ε = 15,900) )

Physical properties of IPQ-3,9-DME 1) Melting point: 250 ° C. or more (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3234 ^{br, w} , 1722 ^s , 1662 ^s , 1444 ^m , 1257 ^vs , 931 ^w 3) ¹ H-NMR spectrum (δ value, ppm): 3.95 (s, 3H),
4.14 (s, 3H), 7.41 (d, J = 2Hz, 1H), 8.62 (s, 1H), 9.29 (s, 1
H), 12.75 (br, 1H) 4) Visible / ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 254, 278, 416

Physical properties of IPQ-1,9-DME 1) Melting point: 300 ° C. or more (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3431 ^{br, m} , 3271 ^{br, w} , 2980 ^w , 1714 ^s , 1645 ^vs , 125
1 ^vs , 995 ^w 3) ¹ H-NMR spectrum (δ value, ppm): 3.99 (s, 3H),
4.20 (s, 3H), 7.43 (d, J = 2Hz, 1H), 8.41 (s, 1H), 9.57 (s, 1
H), 13.53 (br, 1H) 4) Visible and ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 255, 276, 417

Physical properties of IPQ-9-ME 1) Melting point: 300 ° C. or more (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3380 ^{br, m} , 3172 ^w , 2985 ^m , 1705 ^s , 1639 ^vs , 1497 ^m ,
1255 ^vs , 998 ^m 3) ¹ H-NMR spectrum (δ value, ppm): 3.91 (s, 3H),
7.37 (d, J = 2Hz, 1H), 8.62 (s, 1H), 9.68 (s, 1H), 13.89 (b
r, 1H) 4) Visible and ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 257, 278, 417

Physical properties of IPQ-3-ME 1) Melting point: 280 ° C. or more (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3420 ^{br, m} , 3155 ^w , 1722 ^s , 1626 ^s , 1591 ^s , 1444 ^s ,
1232 ^vs , 939 ^m 3) ¹ H-NMR spectrum (δ value, ppm): 4.06 (s, 3H),
7.28 (d, J = 2Hz, 1H), 8.33 (s, 1H), 9.23 (s, 1H), 13.19 (b
r, 1H) 4) Visible / ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 255, 277, 421

Physical properties of IPQ-TAE 1) Melting point: 177 to 179 ° C. or higher 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3160 ^m , 2920 ^s , 1700 ^s , 1650 ^s , 1410 ^m , 1360 ^m , 1
300 ^m , 1210 ^vs , 1085 ^m , 970 ^m , 920 ^m , 850 ^w , 75
0 ^w , 640 ^w 3) ¹ H-NMR spectrum (δ value, ppm): 4.8-6.3 (m, 15
H), 7.31 (d, J = 1.5 Hz, 1H), 8.39 (s, 1H), 9.25 (s, 1H), 1
2.50 (br, 1H) 4) Visible / ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 258 (ε = 20,300), 277 (16,400), 414 ^sh (10,700), 432
(ε = 12,900)

Physical properties of IPQ-9-AE 1) Melting point: 250 ° C. or higher 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3140 ^w , 1705 ^s , 1620 ^m , 1590 ^m , 1230 ^vs , 1180 ^m , 99
0 ^m 3) ¹ H-NMR spectrum (δ value, ppm): 4.86 (m, 2H),
5.3-5.6 (m, 2H), 5.9-6.3 (m, 1H), 7.25 (s, 1H), 8.25 (s, 1
H), 9.24 (s, 1H), 13.17 (br, 1H) 4) Visible and ultraviolet spectrum (λ _max value, nm): (in ethanol) 253, 277, 406

Physical properties of IPQ-TSE 1) Melting point: 105-110 ° C. 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 2920 ^vs , 2850 ^s , 1714 ^s , 1657 ^s , 1254 ^s 3) ¹ H-NMR spectrum (δ value, ppm): 0.83 (t-like, 9
H), 1.24 (m, 90H), 1.88 (m, 6H), 4.52 (m, 6H), 7.57 (s, 1
H), 8.42 (s, 1H), 12.73 (br, 1H) 4) Visible / ultraviolet spectrum (λ _max value, nm): (in ethanol) 258, 278, 415 ^sh , 434

Physical properties of IPQ-9-SE 1) Melting point: 250 ° C. 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3150 ^w , 2920 ^s , 2850 ^s , 1705 ^s , 1620 ^m , 1590 ^s , 12
40 ^vs 990 ^w 3) ¹ H-NMR spectrum (δ value, ppm): 0.87 (t-like, 3
H), 1.26 (m, 32H), 4.41 (m, 2H), 7.36 (s, 1H), 8.55 (s, 1
H), 9.42 (s, 1H), 13.20 (br, 1H) 4) Visible and ultraviolet spectrum (λ _max value, nm): (in ethanol) 253, 277, 412

Physical properties of IPQ-1,3-DIPE 1) Melting point: 300 ° C. (decomposition) 2) IR spectrum (ν _max value, cm ⁻¹ ): (KBr) 3500 ^{br, m} , 3263 ^w , 2985 ^m , 1705 ^s , 1653 ^s , 1308 ^m ,
1255 ^s , 1230 ^vs , 1101 ^s , 831 ^w , 764 ^w , 648 ^w 3) ¹ H-NMR spectrum (δ value, ppm): 1.46 (d, J = 6 Hz,
6H), 1.47 (d, J = 6 Hz, 6H), 5.36 (m, 2H), 6.79 (s, 1H), 8.2
9 (s, 1H), 9.18 (s, 1H), 4) Visible / ultraviolet spectrum (λ _max value, nm): (in acetonitrile) 251,274 ^sh , 428

The IPQs and esters thereof of the present invention can be in any of oral and parenteral dosage forms. In the case of oral administration, it can be administered in usual dosage forms such as capsules, tablets, powders and the like. In the case of parenteral administration, it is administered as an injection, an infusion or the like. In addition, sustained release agents are also effective. To formulate the active ingredient of the present invention, additives such as a surfactant, an excipient, a coloring agent, a preservative, and a coating aid are appropriately used. Also, it can be used in combination with other drugs. The dose and the number of administrations are
Depending on symptoms, age, body weight, dosage form, etc., when administered orally, usually 1 to 1 per day for adults
500mg, 0.1-100m for parenteral administration
g can be administered once or in several divided doses.

[Acute toxicity and nephrotoxicity tests of PQQ and IPQs] (1) Acute toxicity test 1) SPF-ICR mice Male, 5 weeks old (Charles River Japan Co., Ltd.) were given PQQ.Na2 and IPQ.
Were administered intraperitoneally at 20, 40, 80, 160 and 200 mg / kg mouse for 14 days at 25 ° C.
Reared. As IPQs, IPQ, 1-methylpropyl IPQ, 2-methylthioethyl IPQ and benzyl IPQ were used. In addition, one group consisted of 8 animals. As a result, administration of PQQ.Na2 20 mg / kg and 40 mg
No mice died with the / kg administration, but all 5 mice died at the 80 mg administration and all 8 mice died at the 160 mg and 200 mg administrations. Of PQQ · Na ₂ LD ₅₀ is about 70mg /
kg mice. On the other hand, no mice died in IPQs. 2) SPF-ICR mouse Male, 5 weeks old (Charles River Japan Co., Ltd.) was intraperitoneally injected with 0.1 g, 0.2 g, 0.4 g, 0.8 g, or 1.2 g of IPQ per 1 kg of mouse. They were administered and kept at 25 ° C. for 14 days. In addition, one group consisted of 8 animals. IPQ0.1 ~
All mice did not die at 0.4 g and 0.8 g
Then, two died at 1.0 g, and six died at 1.2 g. The LD ₅₀ was about 1.0 g / kg mouse. 3) SPF-ICR mouse Male, 5 weeks old (Charles River Japan Co., Ltd.) was orally administered with 1.0 g, 1.5 g or 2.0 g of IPQ per 1 kg of mouse, and bred at 25 ° C. for 14 days. did. One group consisted of 8 animals. All mice did not die. From the results of 1) to 3), IPQs had significantly lower toxicity than PQQ.

(2) Nephrotoxicity 1) Nephrotoxicity by urinalysis PQQ.Na ₂ and IP
Qs were intraperitoneally administered and mice were bred. Every day, mouse urine is collected and used as a clinical test reagent (trade name: Ulistex I)
I, manufactured by Miles Sankyo Co., Ltd.). As shown in Table 1, sugar was detected in urine of mice to which PQQ.Na ₂ was administered, but no sugar was detected in urine of mice to which IPQs were administered. That is, PQQ showed nephrotoxicity, but IPQs did not.

[0043]

[Table 1]

2) Nephrotoxicity by blood test (a) PQQ.Na ₂ and IPQs were intraperitoneally administered in the same manner as in the acute toxicity test, and mice were bred. One day after administration, the animals were fasted (water was given), and blood was collected 18 hours later to obtain serum. Serum glucose, urea nitrogen and creatinine are used as clinical test reagents (trade name)
Fuji Dry Chem Slide, manufactured by Fuji Photo Film Co., Ltd.). In addition, each value was shown by the average value of eight animals. Table 2 shows the results. In PQQ ・ Na ₂ administration,
A significant decrease in glucose, a significant increase in urea nitrogen and creatinine were observed, and nephrotoxicity was observed. On the other hand, in the administration of IPQs, the respective contents of glucose, urea nitrogen and creatinine were not much different from those in the case of "no administration".

[0045]

[Table 2]

(B) In the same manner as in the acute toxicity test,
150mg, 300mg, 400mg or 60
The mice were intraperitoneally administered at 0 mg / kg, and the mice were bred for one day. Thereafter, fasting (with water) was given, and blood was collected 18 hours later to obtain serum. Serum glucose, urea nitrogen and creatinine are used as clinical test reagents (trade name)
Fuji Dry Chem Slide, manufactured by Fuji Photo Film). In addition, each value was shown by the average value of eight animals. Table 3 shows the results. At any dose of IPQ, the contents of glucose, urea nitrogen and creatinine were not much different from those of "no administration".

[0047]

[Table 3] (B) From the results of (B), nephrotoxicity of PQQ was observed, but nephrotoxicity of IPQs was not observed.

[0048]

The following are examples showing the active oxygen scavenging action of the IPQs and their esters according to the present invention and the preventive and therapeutic actions against various disorders caused by active oxygen. The present invention is not limited to these examples. However, the present invention is not limited to this. Scavenging activity hypoxanthine superoxide Example 1 IPQ class - xanthine O was produced in the system of oxidase ₂ ^- spin trap method using electron spin resonance (hereinafter abbreviated to ESR) the scavenging activity of IPQ acids for (Spin 5,5-dimetyl-1-pyrroline-N-oxide as a trapping agent
(Hereinafter referred to as DMPO)). The following reagents were sequentially placed in a test tube, and hypoxanthine in 7) was added and stirred to start the reaction, which was transferred to a quartz cell for ESR measurement. One minute after the start of the reaction, the ESR was measured and the O ₂ produced
^The drug concentrations (IC ₅₀ values) that reduced-by １ ／ were compared. Incidentally, the drug as, using ascorbic acid as IPQ acids and control, respectively, O ₂ at 5 different concentrations ^- IC ₅₀ values were calculated investigated scavenging activity.

Reaction solution 1) 50 mM sodium phosphate buffer (pH 7.3) 50 μl 2) 10 mM diethylenetriamine pentaacetic acid (hereinafter abbreviated as DTPA) 10 μl 3) xanthine oxidase (2 u / ml) 20 μl 4) distilled water 50 μl 5) Drug 50 μl 6) 900 mM DMPO 10 μl 7) 7.3 mM hypoxanthine 10 μl ESR measurement conditions Magnetic field 236.1 ± 5 mT Output 10 mW Modulation 100 kHz, 0.079 mT Response time 0.3 sec Sweep time 2 min (10 mT interval) Amplification ratio 500 Table 4 shows the results.

[0050]

[Table 4] The IC ₅₀ values of IPQs were 1/5 to 1/20 as compared to the IC ₅₀ value of ascorbic acid. O ₂ of IPQ class - the scavenging activity 5-20 times a strong O ₂ ascorbic acid - had scavenging activity.

Example 2 Hydroxy radical scavenging activity of IPQs The scavenging activity of IPQs against .OH produced in a system of H ₂ O ₂ ─── (Fe ²⁺ -DTPA) was determined in the same manner as in Example 1. Was considered. ^{_{H 2 O 2 + Fe 2+ -DTPAcomplex}} → · OH + OH - + Fe 3+
Was added stirring of H ₂ O ₂ -DTPAcomplex 6 reagent placed in sequential test tube below), the reaction was started and was transferred to ESR measurement quartz cell. One minute after the start of the reaction, the ESR was measured,
The drug concentrations (IC ₅₀ values) for the two concentrations were compared. In addition,
Using IPQs as a drug and ascorbic acid as a control, the OH scavenging activity was examined at five concentrations, and the IC ₅₀ value was calculated. Reaction solution 1) 50 mM sodium phosphate buffer (pH 7.3) 50 μl 2) 1 mM DTPA-Fe ²⁺ complex 10 μl 3) distilled water 70 μl 4) drug 50 μl 5) 900 mM DMPO 10 μl 6) 10 mM H ₂ O ₂ 10 μl ESR measurement conditions field 336 ± 5 mT output 10mW modulation 100kHz, (distance 10 mT) 0.1 mT response time 0.1 seconds sweep time 1 minute amplification ratio 125 Table 5 shows the results.

[0052]

[Table 5] The IC ₅₀ values of IPQ class were almost the same as an IC ₅₀ value of ascorbic acid. IPQs had strong .OH erasing activity like ascorbic acid.

Example 3 A collagenase reflux method (Seglen PO, Methods Cell Biol., 13, 29-8) was obtained from Wistar rats (male, 7 weeks old, weighing about 220 g, purchased from Charles River Japan).
3 (1976)). The cells were 1.4 × 1
0 ⁶ Co / ml so that, Krebs-Ringer solution suspended in the cell fluid 100 [mu] l, 200 mM with a solution in DMSO
Of CCl ₄ solution in 50 μl and 10 μM of lucigenin (Ald
rich, USA), 100 μl, drug solution 50 μl and Kre
Add 700 μl of bs-Ringer solution and set the total amount to 1 to 3
Reaction was performed at 7 ° C for 20 minutes, and chemiluminescence was performed.
luminometer (Laboratorium Berthold, Model LB 950
5, Germany), and the amount of chemiluminescence during the reaction for 20 minutes was measured. As a drug solution, IPQ or 2-methylthioethyl IPQ was added to DMSO at 500 μg /
ml, and the control was DMSO
Was used. IPQ or 2-methylthioethyl IPQ
The light emission amount was reduced to 56.3 ± 0.6% and 57.9 ± 2.1%, respectively, as compared with no addition by adding. The experiment was performed three times. IPQ and 2-methylthioethyl IPQ was erased radicals derived from CCl ₄ strongly. Krebs-Ringer solution 120 mM NaCl, 4.8 mM KCl, 11 mM
Glucose, 0.54mM CaCl ₂ · 4H ₂ O and 1.2mM MgSO ₄ · 7H ₂ O containing 15.
6 mM phosphate buffer.

Example 4 ICR mice (purchased from Charles River Japan Co., Ltd.), males, 70 mice of 6 weeks old were prepared, and 7 groups of 10 mice (A to
G). All mice were fasted for 16 hours and C
The group received 1.75 mg / PQ dissolved in saline.
1.75m dissolved in olive oil for group Q, Na2, D
g / ml of coenzyme Q10, group E: 1.75 mg / ml α-tocopherol dissolved in olive oil, group F: 1.75 mg / ml IPQ, G dissolved in saline.
Groups contained 1.75 mg / ml IP dissolved in olive oil
Q-9-ME was intraperitoneally administered in an amount of 0.2 ml each. The dose is 10 mg / kg mouse each.
45 minutes after administration, Lipopolysaccharide W (E. coli 0111:
B4, manufactured by Sigma) was intraperitoneally administered to all the mice in groups BG in 0.2 ml increments. The dose is 20 mg as LPS.
/ Kg mouse (2.1 mg / kg mouse as lipid A). Thereafter, all mice were fed a commercial solid feed and kept at 25 ° C. for 3 days. Table 6 shows the death process and the survival rate of the mice. IPQs are PQQ ・ N
It can be seen that there is a remarkable preventive effect on endotoxin shock as compared with a ₂ , coenzyme Q ₁₀ and α-tocopherol.

[0055]

[Table 6]

Example 5 IPQs against carbon tetrachloride (CCl ₄ ) -induced liver injury
35 pharmacologically effective SD rats (male, 7 weeks old, weighing about 240 g, purchased from Charles River Japan Co., Ltd.) were prepared and divided into 7 groups (A to G) of 5 rats. All rats were fasted for 16 hours, PQQ.Na2 in group C, IPQ in group D, 1 in group E.
-Methylpropyl IPQ, 2-methylthioethyl IPQ for group F, benzyl IPQ for group G, 5 mg / kg each
Rat (1.2mg of 1.0mg / ml solution administered)
The test compound was administered intraperitoneally in such a way that In addition, 1.1 ml of physiological saline was intraperitoneally administered to the groups A and B instead of the test compound. In addition, 20% after 10 minutes,
CCl4 solution (dissolved in olive oil) was administered 2.4ml by one into the stomach (1 ml / kg rat as CCl _4). A
The rats in the group did not receive the test compound and CCl ₄ . Twenty-four hours after administration of CCl ₄ , all rats were bled from the abdominal aorta and centrifuged to obtain serum. GPT, GOT and total bilirubin levels in serum were measured using Fuji Dry Chem slide. Table 7 shows the results. In addition, each value is an average value of five rats. GPT increased by CCl ₄ administration,
Both GOT and total bilirubin levels were significantly reduced by the administration of the IPQ compound, similarly to the PQQ administration.
It is understood that the PQ compound has an effect of suppressing liver damage.

[0057]

[Table 7]

EXAMPLE 6 IPQ Compound against D-Galactosamine-Induced Liver Injury
35 pharmacologically effective SD rats (male, 7 weeks old, weighing about 240 g, purchased from Charles River Japan Co., Ltd.) were prepared and divided into 7 groups (A to G) of 5 rats. All rats were fasted for 16 hours, PQQ.Na _{2 in} group C, IPQ in group D, 1 in group E.
-Methylpropyl IPQ, 2-methylthioethyl IPQ for group F, benzyl IPQ for group G, 5 mg / kg each
Rat (1.2mg of 1.0mg / ml solution administered)
The test compound was administered intraperitoneally in such a way that In addition, 1.1 ml of physiological saline was intraperitoneally administered to the groups A and B instead of the test compound. Further, 20 minutes later, rats containing the D-galactosamine (0.2 ml) were added to the rats of groups B to G in an amount of 1.2 ml (D
-1 g / kg rat as galactosamine) was administered subcutaneously. Group A rats received no IPQ compound and no D-galactosamine. Twenty-four hours after administration of D-galactosamine, blood was collected from the abdominal aorta of all rats, and serum was obtained by centrifugation. GPT, GOT and total bilirubin levels in serum were measured using Fuji Dry Chem slide. Table 8 shows the results.
In addition, each value is an average value of five rats. D
-GPT, GOT increased by administration of galactosamine
Is decreased by the administration of the IPQ compound in the same manner as the administration of the PQQ, indicating that the IPQ compound has a liver damage suppressing effect.

[0059]

[Table 8]

Example 7 IPQ-9-M against carbon tetrachloride (CCl ₄ ) -induced liver injury
Pharmacological effects of class E (1) SD rats (male, 7 weeks old, weight about 220 g, Japanese charcoal)
25 animals (purchased from Luz River Co., Ltd.) were prepared and divided into five groups (A to E). All rats were fasted for 16 hours, 1.1 ml of a solution containing 0.4 mg / ml of IPQ-9-ME in group C, and 1.0 m of IPQ-9-ME in group D.
g / ml of the solution containing 1.1 ml and IPQ-9-
1.1 ml of a solution containing 2.0 mg / ml of ME was orally administered to each rat, and 40 minutes later, the same amount of IPQ-9-ME was orally administered to each rat. Note that B
The group received 1.1 ml of physiological saline intraperitoneally instead of the IPQ-9-ME solution. Twenty minutes later, rats in groups B to E were intraperitoneally administered with 10% CCl ₄ solution in 2.2 ml portions. Group A rats have IPQ-9-ME and
No CCl ₄ was administered. IPQ-9-ME and C
Cl ₄ was used after being dissolved in olive oil. Twenty-four hours after administration of CCl ₄ , all rats were bled from the abdominal aorta and centrifuged to obtain serum. GPT, GOT and total bilirubin levels in serum were measured using Fuji Dry Chem slide. Table 9 shows the results. In addition, each value is an average value of five rats. CC
The l ₄ administration, increased GPT, both GOT and total bilirubin amount by IPQ-9-ME administration have decreased significantly, IPQ-9-ME is seen that there is a hepatopathy suppressing effect.

[0061]

[Table 9]

Example 8 IPQ-9-M against D-galactosamine-induced liver injury
Pharmacological effects of class E SD rats (male, 7 weeks old, weight about 240 g, Japanese char
30 animals (purchased from Luzliver Co., Ltd.) were prepared and divided into 6 groups (A to F) of 5 animals each. All rats were fasted for 18 hours, Group C contained 1.2 ml of a solution containing 0.4 mg / ml of IPQ-9-ME, and Group D contained 1.0 mg of IPQ-9-ME.
/ Ml, 1.2 ml of a solution containing 2.0 mg / ml of IPQ-9-ME in group E and IP in a group F.
1.2 ml of a solution containing 3.0 mg / ml of Q-9-ME,
Each rat was intraperitoneally administered, and 40 minutes later, the same amount of IPQ-9-ME was intraperitoneally administered.
For group B, physiological saline (1.2 ml) was treated with IPQ-9-M.
Oral administration was performed instead of solution E. After another 20 minutes, B ~
0.2 mg / ml of D-galactosamine was added to rats in group F.
1.2 ml of the solution was subcutaneously administered. Group A rats did not receive IPQ-9-ME and D-galactosamine. Twenty-four hours after the administration of D-galactosamine, all rats were bled from the abdominal aorta and centrifuged to obtain serum. GPT, GO in serum
T and total bilirubin levels were measured using Fuji Dry Chem slide. Table 10 shows the results. In addition, each value is an average value of five rats. Both GPT and GOT increased by administration of D-galactosamine were decreased by administration of IPQ-9-ME,
It can be seen that 9-ME has a liver injury suppressing effect.

[0063]

[Table 10]

Example 9 Drugs of IPQs for hydrocortisone-induced hen egg cataract
Physical effect White leghorn fertilized egg temperature 37 ° C., and an incubator in humidity of 70% of the incubator. The start date of the eggs was 1 day old, and 25 eggs of 15 days old were prepared and divided into 5 groups (A to E). For groups B to E, a small hole was made in the eggshell upper shell of the chicken eggs, and 0.25 μmol / 0.2 ml of sodium hydrocortisone succinate was administered into the air chamber, and then sealed with cellophane tape. To the groups C to E, the following drugs were administered at 1 μmol / 0.2 ml at 3, 10 and 20 hours after the administration of hydrocortisone sodium succinate. Group C was prepared by dissolving PQQ-Na ₂ in distilled water, Group D was prepared by dissolving IPQ in a 10 mM NaHCO ₃ solution, and Group E was prepared by dissolving 10% of methyl IPQ.
Each of them dissolved in a mM NaHCO ₃ solution was administered. 4 hours after administration of hydrocortisone sodium succinate
Eight hours later, the lens is removed and the method of Nishigori et al. (Nishi
gori et al., Inbestigative Ophthalmology & Visual Sci
ence, Vol. 25, p. 1051, 1984). Criteria I: No turbidity in lens. II: There is a faint opaque ring. III: There is a clear cloudy ring. IV: Pinho size opacity is in the nucleus. V: The whole nucleus is cloudy. Table 11 shows the results.

[0065]

[Table 11] IPQs clearly showed a cataract-improving effect similarly to PQQ. It is considered that IPQs have eliminated active oxygen derived from hydrocortisone.

[0066]

Industrial Applicability Since IPQ and its ester compound have a strong active oxygen scavenging effect, they are used as preventive and therapeutic agents for various diseases caused by active oxygen.

Claims (1)

[Claims] 1. A compound of the general formula (I) [Where R is hydrogen, carbamoyl group, phenyl group, 4
A methylphenyl group, an alkyl group having 1 to 4 carbon atoms or a hydroxyl group, a carboxyl group, a carbamoyl group, a methylmercapto group, a phenyl group, a 4-hydroxyphenyl group,
1 carbon atom substituted by a mercapto group, imidazolyl group, amino group, guanidino group, methoxy group or phosphate group
And 4 to 4 alkyl groups. Provided that R ¹ , R ² and R ³ represent hydrogen, an alkyl group, an allyl group, a propargyl group, a benzyl group, a phenyl group or an alkoxycarbonylmethyl group] as an active oxygen-containing compound. Eraser.