JP2007291075A - New compound sterenin and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new improving medicine for metabolic syndrome. <P>SOLUTION: The invention relates to the compound represented by general formula (I) or its salt having 11β-HSD1 inhibiting activity and to the method for producing the compound, [in the formula, R is -CH<SB>2</SB>CH<SB>2</SB>OH, -CH(CO<SB>2</SB>H)CH<SB>2</SB>CH<SB>2</SB>CO<SB>2</SB>H, -H, or -CH<SB>2</SB>COOH]. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to 11β- effective as a therapeutic and / or prophylactic agent for at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, hypertension and metabolic syndrome in which these pathological conditions are combined. Hydroxysteroid dehydrogenase type 1 (hereinafter also referred to as “11β-HSD1”) inhibitory action, and a novel compound that can be used as a raw material for the synthesis of related derivatives derived for the purpose of enhancing the action, and using the compound as an active ingredient The present invention relates to a drug containing the same, a method for producing the compound, and a novel microorganism that produces the compound.

  In recent years, with changes in diet, lifestyle, and living environment, obesity, hyperglycemia, hyperlipidemia, hypertension, and metabolic syndrome patients with these multiple risk factors are increasing worldwide. (Non-Patent Document 1). In any treatment, dietary therapy or exercise therapy is used first, and drug therapy is used in combination when the effect is small or severe.

  Diabetes is classified as insulin-dependent (type I, IDDM) and non-insulin-dependent (type II, NIDDM), with more than 90% of diabetics being the latter. Insulin injection is used for the treatment of IDDM, and for the treatment of NIDDM, a sulfonylurea agent that promotes insulin secretion, a thiazolidinedione agent that improves insulin resistance, a glycosidase inhibitor that inhibits digestive absorption of sugar, Biguanide drugs that suppress gluconeogenesis are used (Non-patent Document 2). However, all drugs do not always have a sufficient effect, and their effectiveness decreases with long-term use, so that an increasing number of patients have serious complications.

  In the treatment of obesity, a central feeding inhibitor is used in severe patients, but due to the limited use period and rebound, a sufficient effect has not been obtained.

  Antihypertensive drugs such as calcium antagonists that dilate blood vessels, diuretics that promote salt excretion, beta-blockers that suppress sympathetic nerves and lower heart rate, alpha-blockers that diminish sympathetic nerves and dilate peripheral blood vessels And angiotensin converting enzyme inhibitors for suppressing vasoconstriction by angiotensin and angiotensin receptor antagonists. However, it is difficult to control blood pressure in the early morning, which is the most frequent stroke period, and the current state of antihypertensive therapy is still not sufficient.

  The drugs for treating hyperlipidemia include HMG-CoA reductase inhibitors that inhibit cholesterol synthesis in the liver, fibrates that inhibit synthesis of neutral fat in the liver, and anion exchange that promotes bile acid excretion. Resin or the like is used. The ultimate goal of hyperlipidemia treatment is to prevent arteriosclerotic diseases such as coronary artery disease and cerebral infarction, but arteriosclerosis is not limited to hyperlipidemia but also hypertension, diabetes, obesity, Since various risk factors such as age develop in an overlapping manner, it is important to always pay attention to these other risk factors, and a multifaceted approach is required (Non-patent Document 3).

Glucocorticoids (cortisol in humans, corticosterone in rodents) release amino acids from muscle through the expression of various proteins and promote the release of fatty acids and glycerol from adipose tissue into the blood. It promotes gluconeogenesis in the liver using a substrate, which in turn promotes an increase in blood sugar. In addition, immature adipocytes in adipose tissue are matured to lead to obesity, and the kidney has the activity of acting on mineralocorticoid receptors to increase blood pressure. In addition to the control by production and secretion in the hypothalamus-pituitary-adrenocortical pathway, the activity control mechanism by glucocorticoid includes recycling by 11β-HSD1 in target organs such as liver, adipose tissue, and lung (non-inhibition). There is a mechanism of conversion from an active form to an active form (Non-patent Document 4). 11β-HSD1 inhibitor suppresses the action of glucocorticoids in these tissues, thereby suppressing hyperglycemia, obesity, hyperlipidemia, and / or hypertension, and exerting multifaceted effects on metabolic syndrome Is expected to do. In adipose tissue-specific 11β-HSD1 highly expressing mice, visceral fat type obesity, impaired glucose tolerance, insulin resistance and increased blood pressure have been reported and support these possibilities (Non-Patent Document 5 or Non-Patent Document 5). Reference 6). As such substances, aminothiazolsulfonamide derivatives (Human Literature 7) and triazole derivatives (WO03 / 065983) are known, but have not yet been put into practical use.
“Nature”, 2001, vol. 414, p. 782-787. “The American Journal of Medicine”, 2000, 108, 238-245 “The Journal of Clinical Endocrinology & Metabolism”, 2004, Vol. 89, p. 2601-2607 “Endocrinology”, 2001, vol. 142, p. 1371-1376 “Science”, 2001, Vol. 294, p. 2166-2170 "The Journal of Clinical Investigation", 2003, Vol. 112, p. 83-90 “Journal of medicinal chemistry”, 2002, 45, p. 3813-3815

  The present invention relates to 11β effective as a therapeutic and / or prophylactic agent for at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, hypertension and metabolic syndrome in which these pathological conditions are combined. -A novel compound that has an inhibitory action on hydroxydehydrogenase type 1 (hereinafter also referred to as "11β-HSD1") and can be used as a raw material for the synthesis of related derivatives derived for the purpose of enhancing these actions, and the compound as an active ingredient It is an object of the present invention to provide a medicament containing the compound, a method for producing the compound, and a novel microorganism that produces the compound.

The present invention includes:

(1) The following general formula (I):

[Wherein, R is —CH ₂ CH ₂ OH
_{-CH (CO 2 H) CH 2} CH 2 CO 2 H
-H or
Or a salt thereof -CH ₂ COOH],
(2) A compound or salt thereof having the following physicochemical properties.
1) Substance properties: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₅ NO ₇
4) Molecular weight: 427 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 450.1526
Calculated value: 450.1529
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
212 nm (ε58600), 265 nm (ε24400), 297 nm (ε10900)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3357,3231,2973,2931,1662,1623,1602,1456,1376,1311,1255,1199,1164,1084,1064,1035,990cm ^-1
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.56 (3H, s), 2.60 (3H, s), 3.37 (2H, m), 3.75 (2H, t, J = 5.1 Hz) , 3.83 (2H, t, J = 5.1 Hz), 4.58 (2H, s), 5.03 (1H, m), 6.23 (1H, br, s), 6.31 (1H , Br, s), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 46.5 (t), 50.9 (t), 61.3 (t), 102 0.0 (d), 105.3 (s), 110.1 (d), 113.1 (d), 122.7 (d), 127.1 (s), 127.2 (s), 132. 7 (s), 133.3 (s), 145.1 (s), 151.3 (s), 152.2 (s), 164.9 (s), 167.1 (s), 170.7 (S), 171.8 (s) ppm,
(3) A compound or salt thereof having the following physicochemical properties.
1) Properties of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₆ H ₂₇ NO ₁₀
4) Molecular weight: 513 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 514.1697
Calculated value: 514.1713
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
214 nm (ε58100), 265 nm (ε26500), 296 nm (ε11100)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3348, 3264, 2973, 2928, 2858, 1718, 1662, 1624, 1603, 1450, 1413, 1377, 1310, 1254, 1198, 1162, 1087, 1063, 1035, 991 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.52 (3H, s), 1.57 (3H, s), 2.25 (1H, m), 2.40 (2H, m), 2.49 (1H, m), 2.60 (3H , S), 3.39 (2H, m), 4.46 (1 H, d, J = 16.9 Hz), 4.58 (1 H, d, J = 16.9 Hz), 4.99 (1 H, m ), 5.04 (1H, m), 6.24 (1H, br, s), 6.31 (1H, br, s), 7.09 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 26.3 (t), 31.9 (t), 46.9 (t), 55 .5 (d), 102.0 (d), 105.3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127. 5 (s), 132.0 (s), 133.3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (S), 171.3 (s), 171.8 (s), 173.9 (s), 176.3 (s) ppm,
(4) A compound or salt thereof having the following physicochemical properties.
1) Properties of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₁ H ₂₁ NO ₆
4) Molecular weight: 383 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 406.1273
Calculated value: 406.1267
6) UV absorption spectrum The UV absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε66700), 268 nm (ε23000), 297 nm (ε12000)
7) Infrared absorption spectrum The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3409, 3267, 2974, 2931, 1669, 1653, 1623, 1599, 1504, 1446, 1413, 1374, 1349, 1313, 1255, 1193, 1173, 1161, 1100, 1063, 1038, 998, 989 cm ^−1
8) 1 H- in nuclear magnetic resonance spectrum heavy methanol, the residual proton signals of (3.31 ppm) was measured as an internal standard ¹ H- nuclear magnetic resonance spectrum is as shown below.
1.50 (3H, s), 1.57 (3H, s), 2.60 (3H, s), 3.38 (2H, m), 4.40 (2H, s), 5.03 (1H , M), 6.22 (1H, br, s), 6.30 (1H, br, s), 7.08 (1H, s) ppm
⁹⁾ 13 C-nuclear magnetic resonance spectrum in deutero methanol, ¹³ C-nuclear magnetic resonance spectrum of the methanol-(49.15ppm) was measured as an internal standard is as indicated below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 45.0 (t), 102.0 (d), 105.3 (s), 110 .2 (d), 113.1 (d), 122.7 (d), 127.3 (s), 129.3 (s), 132.5 (s), 133.3 (s), 145. 1 (s), 151.4 (s), 152.6 (s), 164.9 (s), 167.1 (s), 171.8 (s), 173.4 (s) ppm,
(5) Compound or salt thereof having the following physicochemical properties 1) Property of material: colorless acidic fat-soluble material 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₃ NO ₈
4) Molecular weight: 441 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 442.1495
Calculated value: 442.1502
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε96200), 265 nm (ε41300), 297 nm (ε17500)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3366, 3245, 2973, 2929, 2733, 1662, 1624, 1601, 1458, 1448, 1401, 1377, 1311, 1254, 1201, 1165, 1103, 1081, 1035, 995, 946 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.57 (3H, s), 2.61 (3H, s), 3.39 (2H, m), 4.38 (2H, s), 4.54 (2H , S), 5.04 (1H, m), 6.23 (1H, br, s), 6.30 (1H, br, s), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 45.1 (t), 50.3 (t), 102.0 (d), 105 .3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127.5 (s), 132.1 (s), 133. 3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (s), 170.8 (s), 171.8 (S), 172.8 ppm,
(6) The bacterium producing the compound according to any one of (1) to (5) belonging to the genus Stereoum is cultured, and any one of (1) to (5) is obtained from the culture. The method for producing a compound according to any one of (1) to (5), wherein the compound according to (1) is collected.
(7) Sterium sp. (Sterum sp.) SANK21205 strain (FERM BP-10465) or Sterium sp. (Sterum sp.) SANK21105 strain (FERM BP-10464) is cultured, and from the cultures (1) to (5) The method for producing a compound according to any one of (1) to (5), wherein the compound according to any one of (1) to (5) is collected.
(8) A microorganism belonging to the genus Stereoum and producing the compound according to any one of (1) to (5),
(9) Sterum sp. SANK21205 strain (FERM BP-10465),
(10) Sterium sp. SANK21105 strain (FERM BP-10464),
(11) A pharmaceutical composition comprising as an active ingredient the compound according to any one of (1) to (5) or a pharmacologically acceptable salt thereof,
(12) The pharmaceutical composition according to (11), which is a therapeutic and / or prophylactic agent for at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, hypertension and metabolic syndrome,
(13) An 11β-hydroxysteroid dehydrogenase type1 (11β-HSD1) inhibitor comprising the compound according to any one of (1) to (5) or a salt thereof,
About.

The present invention also relates to the compounds described below.
(I) A compound having the following physicochemical properties or a salt thereof.
1) Substance properties: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₅ NO ₇
4) Molecular weight: 427 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 450.1526
Calculated value: 450.1529
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
212 nm (ε58610), 265 nm (ε24385), 297 nm (ε10909),
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3357, 3231, 2973, 2931, 1662, 1623, 1602, 1456, 1376, 1311, 1255, 1199, 1164, 1084. 1064, 1035, 990 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.56 (3H, s), 2.60 (3H, s), 3.37 (2H, m), 3.75 (2H, t, J = 5.1 Hz) , 3.83 (2H, t, J = 5.1 Hz), 4.58 (2H, s), 5.03 (1H, m), 6.23 (1H, s, br), 6.31 (1H , S, br), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 46.5 (t), 50.9 (t), 61.3 (t), 102 0.0 (d), 105.3 (s), 110.1 (d), 113.1 (d), 122.7 (d), 127.1 (s), 127.2 (s), 132. 7 (s), 133.3 (s), 145.1 (s), 151.3 (s), 152.2 (s), 164.9 (s), 167.1 (s), 170.7 (S), 171.8 (s) ppm
(Ii) A compound having the following physicochemical properties or a salt thereof.
1) Properties of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₆ H ₂₇ NO ₁₀
4) Molecular weight: 513 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 514.1697
Calculated value: 514.1713
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
214 nm (ε58133), 265 nm (ε26506), 296 nm (ε11145)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3348, 3264, 2973, 2928, 2858, 1718, 1662, 1624, 1603, 1450, 1413, 1377, 1310, 1254, 1198, 1162, 1087, 1063, 1035, 991 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.52 (3H, s), 1.57 (3H, s), 2.25 (1H, m), 2.40 (2H, m), 2.49 (1H, m), 2.60 (3H , S), 3.39 (2H, m), 4.46 (1 H, d, J = 16.9 Hz), 4.58 (1 H, d, J = 16.9 Hz), 4.99 (1 H, m ), 5.04 (1H, m), 6.24 (1H, s, br), 6.31 (1H, s, br), 7.09 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 26.3 (t), 31.9 (t), 46.9 (t), 55 .5 (d), 102.0 (d), 105.3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127. 5 (s), 132.0 (s), 133.3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (S), 171.3 (s), 171.8 (s), 173.9 (s), 176.3 (s) ppm
(Iii) a compound having the following physicochemical properties or a salt thereof;
1) Properties of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₁ H ₂₁ NO ₆
4) Molecular weight: 383 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 406.1273
Calculated value: 406.1267
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε66746), 268 nm (ε22967), 297 nm (ε11962)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3409, 3267, 2974, 2931, 1669, 1653, 1623, 1599, 1504, 1446, 1413, 1374, 1349, 1313, 1255, 1193, 1173, 1161, 1100, 1063, 1038, 998, 989 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.57 (3H, s), 2.60 (3H, s), 3.38 (2H, m), 4.40 (2H, s), 5.03 (1H , M), 6.22 (1H, s, br), 6.30 (1H, s, br), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 45.0 (t), 102.0 (d), 105.3 (s), 110 .2 (d), 113.1 (d), 122.7 (d), 127.3 (s), 129.3 (s), 132.5 (s), 133.3 (s), 145. 1 (s), 151.4 (s), 152.6 (s), 164.9 (s), 167.1 (s), 171.8 (s), 173.4 (s) ppm
(Iv) A compound having the following physicochemical properties or a salt thereof.
1) Substance properties: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₃ NO ₈
4) Molecular weight: 441 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 442.1495
Calculated value: 442.1502
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε96167), 265 nm (ε41250), 297 nm (ε17500)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3366, 3245, 2973, 2929, 2733, 1662, 1624, 1601, 1458, 1448, 1401, 1377, 1311, 1254, 1201, 1165, 1103, 1081, 1035, 995, 946 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.57 (3H, s), 2.61 (3H, s), 3.39 (2H, m), 4.38 (2H, s), 4.54 (2H , S), 5.04 (1H, m), 6.23 (1H, s, br), 6.30 (1H, s, br), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 45.1 (t), 50.3 (t), 102.0 (d), 105 .3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127.5 (s), 132.1 (s), 133. 3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (s), 170.8 (s), 171.8 (S), 172.8 ppm.

  According to the present invention, the compounds having 11β-HSD1 inhibitory activity, sterinin A, sterinin B, sterinin C and sterinin D, are provided. Furthermore, the present invention provides a microorganism for producing the compound, and a method for producing the compound using the microorganism. The compound of the present invention is useful for the treatment and / or prevention of at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, hypertension and metabolic syndrome in which these pathological conditions are combined.

1. Sterenin A, Sterinin B, Sterinin C, and Sterinin D
In this specification, Sterenin A (Sterenin A) means the following general formula (I):

In which R is —CH ₂ CH ₂ OH or a compound having the physicochemical properties described in (i) above. Similarly, the Suterenin B (Sterenin B), shows the compound R has the physicochemical properties described in _{CH (CO 2 H) CH 2} CH 2 CO 2 H , compound or above (ii), Suterenin C (Sterenin C) refers to a compound in which R is —H or a compound having the physicochemical properties described in (iii) above. Sterinin D (Sterenin D) refers to a compound in which R is —CH ₂ COOH. Or the compound which has the physicochemical property as described in said (iv) is shown.

  The compound of the present invention is a derivative having at least one action selected from the group consisting of a blood glucose level lowering action, an obesity improving action, a blood lipid lowering action, a blood pressure lowering action, and / or a metabolic syndrome improving action. It can also be used as a starting material for the synthesis.

  The compound of the present invention can be converted to a salt by a method well known to those skilled in the art using a base. The present invention also includes these salts. When the salt of the compound of the present invention is used as a medicine, it is not particularly limited as long as it is used medically and is pharmacologically acceptable, and when used for purposes other than medicine, it is used as an intermediate, for example. In that case, there is no particular limitation as long as it can be used for the purpose.

  Examples of such salts include alkali metal salts such as sodium salt, potassium salt and lithium salt; alkaline earth metal salts such as calcium salt and magnesium salt; aluminum salt, iron salt, zinc salt and copper Metal salts such as salts, nickel salts and cobalt salts; inorganic salts such as ammonium salts; t-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglu Camin salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N, N'-dibenzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, N-benzyl-phenethylamine salt, piperazine salt, tetramethylammonium salt , Tris (hydroxy Organic amine salts such as methyl) aminomethane salts; amino acid salts such as glycine salts, lysine salts, arginine salts, ornithine salts, asparagine salts, preferably pharmacologically acceptable salts, more preferably Are sodium, potassium and ammonium salts.

  In addition, the compound of the present invention and a salt thereof may be left in the air or mixed with water or an organic solvent to combine with water or a solvent to form a hydrate or a solvate. Hydrates and solvates of are also included in the present invention.

  Furthermore, since stelenin B has an asymmetric carbon atom, there are two optical isomers. In the general formula (I), these isomers are represented by a single formula, but the present invention includes all of these isomers including racemates, and mixtures of these isomers. These isomers can be produced by a stereospecific synthesis method or by synthesis using an optically active compound as a raw material compound.

2. Microorganisms used in the method for producing Sterenine A, Sterinin B, Sterenin C and Sterenin D As strains belonging to the genus Stereum used in the method for producing Sterenin A, Sterinin B, Sterinin C and Sterenin D, For example, Sterum sp. SANK21205 strain (hereinafter referred to as “SANK21205 strain”), or Sterium sp. SANK21105 strain (hereinafter referred to as “SANK21105 strain”) can be mentioned. The SANK21205 strain was isolated from basidiomycetous basidiomycetes collected in Gunma Prefecture. In addition, SANK21105 strain was isolated from basidiomycetous basidiomycetes collected in Shizuoka Prefecture.

In order to observe the bacteriological properties of the bacterium, culture was performed on the following media. The composition of each medium used is described below.

<PDA medium (Potato Dextrose Agar medium)>
Potato dextrose agar medium "Nissui" (Nissui Pharmaceutical Co., Ltd.) 39g
Agar 5g
1000ml distilled water
<MEA medium (malt extract agar medium)>
Malt extract 12.5g
Agar 20g
1000ml distilled water
The display of the color tone was in accordance with “Metune Handbook of Color” (Kornerup A. & Wanscher J. H. (1978) Method Handbook of Color (3rd. Edition) .Erye Methuen, London).

The mycological properties of SANK21205 strain under culture are as follows.

  Growth on PDA medium is 41 to 49 mm at 23 ° C. for 1 week. The fungus is fluffy and exhibits yellowish white (4A2) in the center. The back side is pale yellow (4A3) in the center. The aerial hyphae are 1.5 to 8 μm wide, thin walls, and smooth. There are 0 to 4 thin connections per partition. Conidia are not formed. Mycelia with insoluble fine oil droplets attached to the center of the fungus are observed. Growth on MEA medium is 34-40 mm at 23 ° C. for 1 week of culture. The fungus is thin and white. There is no change on the back side.

  The mycological properties of SANK21105 strain under culture are as follows.

  Growth on PDA medium is 50 to 56 mm at 23 ° C. for 1 week of culture. The fungus is fluffy and has pastel yellow (3A4) to light yellow (4A4) in the center. The back side is pale yellow (4A3) in the center. The aerial hyphae are 1.5 to 8 μm wide, thin walls, and smooth. There are 0 to 3 faint connections per partition. Mycelia with insoluble fine oil droplets attached to the center of the fungus are observed. Growth on MEA medium is 37 to 42 mm at 23 ° C. for 1 week. The fungus is thin and white. There is no change on the back side.

  In order to identify the SANK21205 strain and the SANK21105 strain, the mycological properties of the dried fruit body specimens used for separation were observed. The mycological properties of the dried specimen are as follows.

  The fruiting body is back-and-back half-back. Umbrellas are 5mm or less in width, 0.4mm or less in thickness, shell-shaped or fan-shaped, side by side, and are firm. The surface is rough and hairy and grayish white. The edge of the umbrella is thin and sharp. The grain layer surface is smooth or slightly humpy and pale orange. A lower skin exists between the coat and the meat. The lower skin has a thickness of 25 μm or less and has a brown color.

  Basidiospores are 8 to 12 × 3 to 4 μm, cylindrical and may be slightly curved, and are thin walls, smooth and amyloid. The basidiode is 40 to 50 x 5 to 6 µm, has a narrow rod shape, and has four spores. The petite is 4 μm long. Pseudo cystidia may have a width of 4.5 to 6 μm, a cylindrical shape, a thick wall (less than 2 μm) at the base, a thin wall at the tip, and a small protrusion at the tip. Mycelium is one mycelium type, width 2 to 6 μm, thin wall to thick wall, and no faint connection is observed in the partition walls. No Acantopeia or Fake Acanthifidia is observed.

The above-mentioned mycological properties are described in Chamris (1988) “Non-spotitate Stereoid Fungiy in the Northeastern United States and Adjustable Canada” J Climer, Berlin (Chamuris GP (1988) The non-stipitate. In accordance with the description of the genus Stereoid of the stereoid fungi in the northeastern united states and adjunct Canada. J. Cramer, Berlin. The SANK21205 strain and the SANK21105 strain are of the same species, and the SANK21205 strain and the SANK21105 strain were identified as Sterum sp.
In addition, SANK21205 stock is STELLIUM SP SANK21205 stock, SANK21105 stock is STELLIUM SP SANK21105 stock, and it is an independent administrative agency National Institute of Advanced Industrial Science and Technology, 1-1-1 East Tsukuba City, Ibaraki Prefecture, Japan on December 6, 2005. Deposited internationally at the Patent Biological Depositary Center and were assigned the deposit numbers FERM BP-10465 and FERM BP-10464, respectively.

  As is well known, fungi are susceptible to mutation in nature or by artificial manipulation (for example, ultraviolet irradiation, radiation irradiation, chemical treatment, etc.), and the same applies to the SANK21205 and SANK21105 strains of the present invention. is there. The SANK21205 strain and the SANK21105 strain referred to in the present invention include all mutants thereof.

  These mutant strains also include those obtained by genetic methods such as recombination, transduction, transformation and the like. That is, the SANK21205 strain and the SANK21105 strain, the mutant strains and strains that are not clearly distinguished from them are all included in the SANK21205 strain and the SANK21105 strain.

3. Method for culturing microorganisms producing Sterenine A, Sterinin B, Sterenin C and Sterenin D, and purification method for Sterenin A, Sterinin B, Sterinin C and Sterenin D (1) Culture method Sterinin A, Sterinin B, Sterinin C and The cultivation of these microorganisms to obtain stellinin D is generally carried out using a medium similar to that used to produce other fermentation products. Such a medium contains a carbon source, a nitrogen source and an inorganic salt that can be assimilated by the microorganism.

Examples of the carbon source include glucose, fructose, maltose, sucrose, mannitol, glycerin, dextrin, oats, rye, barley, brown rice, corn starch, potato, corn flour, soybean oil, cottonseed oil, molasses, citric acid, tartaric acid Etc. These carbon sources can be used alone or in combination.
The amount of the carbon source added to the medium varies depending on other components in the medium, but is usually 1 to 10% by weight of the medium amount.

  Examples of nitrogen sources include soy flour, bran, peanut powder, cottonseed powder, casein hydrolyzate, pharmamine, corn steep liquor, peptone, meat extract, yeast, yeast extract, malto extract, sodium nitrate, ammonium nitrate, Examples thereof include ammonium sulfate and sodium L-glutamate. These carbon sources can be used alone or in combination.

  The amount of nitrogen source added to the medium is usually 0.2 to 6% by weight, although it depends on the other components in the medium.

  A carbon source and a nitrogen source are generally used in combination, but need not be in a pure form, and may be less pure containing trace amounts of growth factors, vitamins and inorganic nutrients.

  Examples of inorganic salts include salts that generate ions such as sodium, potassium, magnesium, ammonium, calcium, phosphoric acid, sulfuric acid, hydrochloric acid, and carbonic acid.

  In addition, vitamin B1, which can be assimilated by bacteria, vitamins such as biotin, bacterial cell growth promoting substances such as thiamine, and the like may be added as necessary.

  Further, manganese, molybdenum, and other metal salts may be added as necessary.

  When the nutrient medium is foamed, an antifoaming agent such as silicone oil, polyalkylene glycol ether, vegetable oil, animal oil, or surfactant may be added to the medium as necessary. It is particularly preferable to add these antifoaming agents during liquid culture.

  The pH of the medium varies depending on the components of the medium, the growth temperature, and the like, but may be any pH that is usually used for producing a fermentation product.

  The growth temperature of the fungus varies depending on the components of the medium, pH, and the like, but is usually 15 ° C. to 30 ° C., and the range of 20 to 28 ° C. is good for growth. 20 to 26 ° C. is suitable for production of Sterenin A, Sterinin B, Sterinin C and Sterinin D.

  The culture method is not particularly limited, and any culture method generally used for microorganisms may be used, and a culture method using a solid medium, a stirring culture method, a shaking culture method, an aeration culture method, or the like can be used. The culture is performed using a solid medium.

  In the culture, solid culture is preferably performed at 20 to 26 ° C., more preferably at 23 ° C. for 10 days or longer. The culture is initiated in an Erlenmeyer flask by a one or two stage seed development process. The medium for the seed development stage can be used in combination with a carbon source, a nitrogen source, a trace amount of growth factors, inorganic salts and trace metals. Seed culture Erlenmeyer flasks are shaken in a rotary shaker at 20 to 26 ° C. (preferably 23 ° C.), 5 to 10 days (preferably 7 days), or shaken until fully grown. . The grown seed culture is used to inoculate the second seed culture medium or the production culture medium. In the case of using an intermediate growth process, the seed culture solution is grown essentially in the same manner as the first seed culture, and the seed culture solution is inoculated into a production culture medium in a solid medium culture vessel such as an Erlenmeyer flask. The inoculated solid medium culture vessel is allowed to stand at a constant temperature for production culture.

(2) Purification method of sterinin A, sterinin B, sterinin C and sterinin D In the case of solid culture after the completion of culturing, selenin A, sterinin B, sterinin C and sterinin D present in the solid culture are 50 to 90%. After extraction with water-containing acetone or water-containing methanol and removing the organic solvent by concentration, an organic solvent that is not miscible with water under acidic conditions, for example, ethyl acetate, chloroform, ethylene chloride, methylene chloride, butanol alone or , And can be extracted and purified by a combination thereof. Or, as an adsorbent, for example, Amberlite XAD-2, XAD-4 (manufactured by Rohm and Haas Co.) which is activated carbon or an adsorbing resin, Diaion HP-10, HP-20, CHP20P, Extraction and purification can also be performed using HP-50, Sepabeads SP-207 (manufactured by Mitsubishi Chemical Corporation), or the like. When extracting and purifying using an adsorbent, the liquid containing the target compound is removed by passing the adsorbent layer and adsorbing the impurity to the adsorbent, or the target compound is adsorbed and the impurity is washed away. Thereafter, the target compound can be extracted and purified by eluting the target compound with methanol water, acetone water, butanol water or the like.

  In the case of liquid culture, Selenin A, Sterinin B, Sterinin C and Sterenin D present in the liquid part and the bacterial body in the culture medium, or both, are all of the culture end liquid, or the bacterial body and other solid parts. Separation by filtration using diatomaceous earth as a filter aid or centrifugation, and the physicochemical properties of the obtained filtrate or supernatant and bacterial cells using 11β-HSD1 inhibitory activity or high performance liquid chromatography as an index. It can be extracted and purified.

  Strainin A, Sterinin B, Sterinin C and Sterinin D present in the filtrate or supernatant are organic solvents immiscible with water, for example, ethyl acetate, chloroform, ethylene chloride, methylene chloride, butanol, etc. It can be extracted by combination. These extracts can be purified by extracting and purifying the target compound by performing the same extraction and purification operation as in solid culture.

  Stellinin A, Stellinin B, Stellinin C and Stellinin D present in the cells are extracted with 50 to 90% water-containing acetone or water-containing methanol, the organic solvent is removed by concentration operation, and extraction and purification operations are performed in the same manner as above. It can be obtained by doing. For example, the target compound can be extracted by adding acetone or methanol in an appropriate amount (preferably a final concentration of 50%) after completion of the culture. After completion of the extraction, the target compound can be extracted and purified by performing a filtration operation using diatomaceous earth as a filter aid and performing the same extraction and purification operation as in the solid culture.

  A solution containing Sterenin A, Sterinin B, Sterinin C and Sterenin D extracted and purified by the above method is silica gel, Sephadex LH-20 (manufactured by Amersham Biosciences), Florisil, Cosmocil (Nacalai Tesque Co., Ltd.) Column chromatography using a carrier such as Diaion HP-20, CHP20P (manufactured by Mitsubishi Chemical Corporation); Sephadex G-10 (Amersham Bioscience) Manufactured by Toyopearl HW40F (manufactured by Tosoh Corporation), etc .; Dowex 1 (manufactured by Dow Chemical Co., Ltd.), Diaion PA316 (manufactured by Mitsubishi Chemical Corporation), DEAE Sepha Dex A-25 (Amersham Bioscience Ion-exchange chromatography using, for example, Ltd.)); and normal phase can be further purified by high performance liquid chromatography using a reversed phase column.

  By using the above separation and purification means alone or in combination as appropriate, and depending on the case, it is possible to separate and purify the sternin A, sterinin B, sterinin C and sterinin D of the present invention.

  As mentioned above, although the representative method of the manufacturing method of Sterenine A, Sterinin B, Sterinin C, and Sterinin D was demonstrated, a manufacturing method is not limited to these, The manufacturing method other than these already known to those skilled in the art is used. You can also.

4). A pharmaceutical comprising, as an active ingredient, at least one compound selected from the group consisting of sterinin A, sterinin B, sterinin C, and stellenin D, or a pharmacologically acceptable salt thereof. At least one compound selected from the group consisting of stellenine D or a pharmacologically acceptable salt thereof can be administered in various forms. Examples of the administration form include oral administration by tablets, capsules, granules, emulsions, pills, powders, syrups (solutions), etc., or injections (intravenous, intramuscular, subcutaneous or intraperitoneal administration), Examples include parenteral administration such as instillation and suppository (rectal administration). These various preparations are usually used in the pharmaceutical preparation technical field such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizers, suspension agents, coating agents, etc. as main ingredients in accordance with conventional methods. It can be formulated with the resulting adjuvant.

  When used as a tablet, as a carrier, for example, excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid; water, ethanol, propanol, simple syrup, glucose Solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc .; dried starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid Disintegrators such as esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose; disintegrators such as sucrose, stearin, cocoa butter, hydrogenated oil; quaternary ammonium salts, sodium lauryl sulfate Moisturizers such as glycerin and starch; Adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid; Uses a lubricant such as purified talc, stearate, boric acid powder and polyethylene glycol be able to. Moreover, it can be set as the tablet which gave the normal coating as needed, for example, a sugar-coated tablet, a gelatin-encapsulated tablet, an enteric-coated tablet, a film-coated tablet, a double tablet, and a multilayer tablet.

  When used as pills, as carriers, for example, excipients such as glucose, lactose, cocoa butter, starch, hydrogenated vegetable oil, kaolin, talc; binders such as gum arabic powder, tragacanth powder, gelatin, ethanol; Disintegrants such as can be used.

  When used as a suppository, a carrier conventionally known in this field can be widely used as a carrier, and examples thereof include polyethylene glycol, cocoa butter, higher alcohol, esters of higher alcohol, gelatin, semi-synthetic glyceride and the like.

  When used as an injection, it can be used as a solution, emulsion or suspension. These solutions, emulsions or suspensions are preferably sterilized and isotonic with blood. The solution used for the production of these solutions, emulsions or suspensions is not particularly limited as long as it can be used as a diluent for medical use. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isopropanol. Examples include stearyl alcohol and polyoxyethylene sorbitan fatty acid esters. In this case, a sufficient amount of sodium chloride, glucose or glycerin may be included in the preparation to prepare an isotonic solution, and a normal solubilizing agent, buffer, soothing agent, etc. may be included. You may go out.

  Moreover, a coloring agent, a preservative, a fragrance | flavor, a flavoring agent, a sweetening agent, etc. can also be included in said formulation as needed, and also other pharmaceuticals can be included.

  The amount of the active ingredient compound contained in the preparation is not particularly limited and is appropriately selected within a wide range, but is usually 1 to 70% by weight, preferably 1 to 30% by weight, based on the total composition.

    The amount used varies depending on symptoms, age, body weight, administration method, dosage form, etc., but is usually 2000 mg (preferably 100 mg) as an upper limit per day for adults, and 0.1 mg (preferably as the lower limit) 1 mg, more preferably 10 mg) can be administered once a day or divided into several times according to symptoms.

Next, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.
(Example 1) Culture of SANK21205 strain About 1 cm square mycelium pieces were cut from the slant of the SANK21205 strain and suspended in a glass potter filled with about 3 ml of sterile water in advance. The mycelium pieces were homogenized using a glass potter and inoculated into a 100 ml Erlenmeyer flask containing 30 ml of a seed culture medium (Table 1) having the composition described below, which was sterilized (121 ° C., 20 minutes). The same operation was performed to prepare 15 seed culture flasks. The cells were cultured for 7 days in a rotary shaker at 210 rpm at 23 ° C. The seed culture solution thus obtained was sterilized (121 ° C., 40 minutes) and a 500 ml solid medium culture vessel (Agriflex, Fujimori Industry Co., Ltd.) containing the medium composition of the production culture medium (Table 2) described later. 15) each was inoculated with 30 ml, left at 23 ° C., and cultured for 14 days.

(Table 1) Medium composition of seed culture medium ―――――――――――――――――――――――――――――――――――――
4g glucose
Malt extract (Difco) 10g
Yeast extract (Difco) 4g
Agar 3g
Defoamer * 0.05ml
1000ml distilled water
―――――――――――――――――――――――――――――――――――――
* Nissan Disform CB-442 (registered trademark, manufactured by NOF Corporation)

(Table 2) Medium composition of production culture medium (per 500ml solid medium culture container)
―――――――――――――――――――――――――――――――――――――
90g brown rice
NaNO3 0.05g
KH2PO4 0.05g
MgSO4 ・ 7H2O 0.05g
100ml distilled water
―――――――――――――――――――――――――――――――――――――

(Example 2) Isolation / purification of sterinin A, sterinin B, sterinin C and sterinin D In the following purification, the active fraction was monitored using HPLC under the following conditions.

Column: Capsule pack (CAPCELL PAK) C ₁₈ UG120:
4.6φ × 150mm (made by Shiseido Co., Ltd.)
Solvent: 35% acetonitrile water containing 0.05% formic acid Flow rate: 1.0 ml / min Detection: UV absorption 210 nm
Retention time: 8.49 minutes (Stellenin A)
9.60 minutes (Stellenin B)
9.65 minutes (Stellenin C)
10.71 minutes (Stellenin D)
Acetone (3 L) was added to the culture end product (3 L) obtained in Example 1 and allowed to stand for 30 minutes with stirring. Celite 545 (150 g) was added to the extract and filtered. The obtained filtrate was adjusted to pH 3 with 6N HCl, and extracted by adding ethyl acetate (2 L). Ethyl acetate (1.5 L) was again added to the obtained aqueous layer for extraction, and the organic layers obtained by the two extraction operations were combined. The organic layer (3.5 L) was washed with saturated brine (2 L), then anhydrous sodium sulfate (400 g) was added and dehydrated for 1 hour. After removing anhydrous sodium sulfate by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain an oily substance (13 g). This oily substance was dissolved in methanol (50 ml), and then added to Cosmosil 140C18 (60 g: manufactured by Nacalai Tesque) with stirring, and the methanol was distilled off. An aqueous solution (400 ml) containing 0.3% triethylamine-phosphoric acid and adjusted to pH 3 was added to and suspended in Cosmosil 140C18 coated with an oily substance, and then containing 0.3% triethylamine-phosphoric acid. To a Cosmosil 140C18 column (600 ml) equilibrated with 10% acetonitrile water adjusted to pH 3. After washing the column with the same solvent (1 L), the column was developed with 20% aqueous acetonitrile (1 L) containing 0.3% triethylamine-phosphoric acid and adjusted to pH 3, and the eluate was fractionated every 17 ml. (Fr. 1 to 62). Then, 25% acetonitrile water (1.6 L, Fr. 63 to 159) adjusted to pH 3 containing 0.3% triethylamine-phosphoric acid, adjusted to pH 3 containing 0.3% triethylamine-phosphoric acid. Developed in the order of 30% acetonitrile water (3 L, Fr. 160 to 335), and Fr. containing Sterinin A, Sterinin B, Sterinin C and Sterinin D as main components. Fractions from 265 to 294 (500 ml) were obtained. The obtained fraction was concentrated to distill off acetonitrile, followed by extraction with ethyl acetate (400 ml). The aqueous layer was extracted again by adding ethyl acetate (300 ml), and the organic layers obtained by the two extraction operations were combined. The organic layer (700 ml) was washed with saturated brine (600 ml), dehydrated with anhydrous sodium sulfate (80 g), and dehydrated for 1 hour. After removing anhydrous sodium sulfate by filtration, the filtrate was concentrated to dryness under reduced pressure to obtain a crude product (335 mg). A part (130 mg) of this crude product was dissolved in methanol (2 ml), and then the solution was divided into 10 times 200 μl and equilibrated with 37% acetonitrile water containing 0.05% formic acid (CAPCELL). PAK C ₁₈ UG120, 20φ × 250 mm (manufactured by Shiseido Co., Ltd.) The column was developed at a flow rate of 10.0 ml / min, ultraviolet absorption at 210 nm was detected, and a retention time of 25.0 minutes ( The active fractions eluted at Sterenine A), 26.5 min (Sterenin B), 29.0 min (Sterenin C) and 30.2 min (Sterenin D) were collected, respectively. The fractions were combined, concentrated separately under reduced pressure, and lyophilized, and then sterinin A (24 mg), sterinin B (25 mg), sterinin C (11 mg) and stellenin D (10 mg) (Test Example) 11β-HSD1 enzyme inhibition experiment (1) Enzyme source cDNA encoding the full length of human 11β-HSD1 (GenBank Accession No .: NM_005525, sequence is described in VERSION: NM_005525.2) Was introduced into HEK293 cells according to the package insert using Lipofectamine plus reagent (Invitrogen), and the cells after 48 hours were collected and frozen at -80 ° C. As a control, cells transfected with the pCIneo vector were similarly prepared by thawing the cells with a buffer containing a final concentration of 20 mM hepes, 1 mM ethylenediaminetetraacetic acid, 2 mM magnesium chloride, and a protease inhibitor cocktail (Roche). Suspended and filled with nitrogen gas The cells were crushed by applying pressure of 4.1 MPa on ice for 30 minutes, and the supernatant was recovered by centrifuging the cell lysate at 4 ° C. for 10 minutes at 1,000 × g. Was further centrifuged at 105,000 × g for 30 minutes at 4 ° C., and the resulting precipitate was suspended as a microsomal fraction in assay buffer (50 mM Tris buffer, 10% glycerol) to obtain an enzyme source.

(2) Enzyme inhibition experiment The reaction was carried out on a 96-well PCR plate (Perkin Elmer) at a reaction dose of 20 μL, and all samples were diluted in assay buffer (50 mM Tris buffer, 10% glycerol). Plate with NADPH at a final concentration of 5 mM, glucose 6-phosphate at 50 mM, glucose 6-phosphate dehydrogenase (Sigma) at 10 units / mL, magnesium chloride at 20 mM, and microsomal fraction at 5 μg protein per well And a solution containing a test compound dissolved in dimethyl sulfoxide (any one of Sterenin A, Sterinin B, Sterinin C and Sterinin D) is added to a final concentration of 0.1%, For 15 minutes. After completion of preincubation, cortisone with a final concentration of 500 nM was added to start the reaction, and the reaction was performed at room temperature for 60 minutes. The plate was heated at 99 ° C. for 10 minutes to stop the reaction, and the amount of produced cortisol was measured with a Discovery (Packard) according to the package insert using a Cortisol prototype kit (CIS Bio International). For the ββ-HSD1-expressing cell-dependent cortisol production, which is not seen in the microsome fraction derived from the control cells treated in the same way, sterenin A inhibited 86% at 10 μM and 61% inhibited at 10 μM. Sterenin C showed 93% inhibition at 10 μM and Sterenin D showed 83% inhibition at 10 μM.

According to the present invention, compounds having 11β-HSD1 inhibitory activity, sterinin A, sterinin B, sterinin C and sterinin D are provided. Furthermore, the present invention provides a microorganism for producing the compound, and a method for producing the compound using the microorganism. The compound of the present invention is useful for the prevention and / or treatment of at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, and hypertension and / or metabolic syndrome that develops these pathological conditions in combination. It is.

Claims (13)

The following general formula (I):


[Wherein, R is —CH ₂ CH ₂ OH
_{-CH (CO 2 H) CH 2} CH 2 CO 2 H
-H or
A compound represented by —CH ₂ COOH] or a salt thereof. Compound or salt thereof having the following physicochemical properties 1) Property of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₅ NO ₇
4) Molecular weight: 427 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 450.1526
Calculated value: 450.1529
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
212 nm (ε58600), 265 nm (ε24400), 297 nm (ε10900)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3357,3231,2973,2931,1662,1623,1602,1456,1376,1311,1255,1199,1164,1084,1064,1035,990cm ^-1
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.56 (3H, s), 2.60 (3H, s), 3.37 (2H, m), 3.75 (2H, t, J = 5.1 Hz) , 3.83 (2H, t, J = 5.1 Hz), 4.58 (2H, s), 5.03 (1H, m), 6.23 (1H, br, s), 6.31 (1H , Br, s), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 46.5 (t), 50.9 (t), 61.3 (t), 102 0.0 (d), 105.3 (s), 110.1 (d), 113.1 (d), 122.7 (d), 127.1 (s), 127.2 (s), 132. 7 (s), 133.3 (s), 145.1 (s), 151.3 (s), 152.2 (s), 164.9 (s), 167.1 (s), 170.7 (S), 171.8 (s) ppm. Compound or salt thereof having the following physicochemical properties 1) Property of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₆ H ₂₇ NO ₁₀
4) Molecular weight: 513 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 514.1697
Calculated value: 514.1713
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
214 nm (ε58100), 265 nm (ε26500), 296 nm (ε11100)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3348, 3264, 2973, 2928, 2858, 1718, 1662, 1624, 1603, 1450, 1413, 1377, 1310, 1254, 1198, 1162, 1087, 1063, 1035, 991 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.52 (3H, s), 1.57 (3H, s), 2.25 (1H, m), 2.40 (2H, m), 2.49 (1H, m), 2.60 (3H , S), 3.39 (2H, m), 4.46 (1 H, d, J = 16.9 Hz), 4.58 (1 H, d, J = 16.9 Hz), 4.99 (1 H, m ), 5.04 (1H, m), 6.24 (1H, br, s), 6.31 (1H, br, s), 7.09 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 26.3 (t), 31.9 (t), 46.9 (t), 55 .5 (d), 102.0 (d), 105.3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127. 5 (s), 132.0 (s), 133.3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (S), 171.3 (s), 171.8 (s), 173.9 (s), 176.3 (s) ppm. Compound or salt thereof having the following physicochemical properties 1) Property of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₁ H ₂₁ NO ₆
4) Molecular weight: 383 (measured by TOF mass spectrum method)
5) The accurate mass [M + Na] ⁺ measured by the high-resolution TOF mass spectrum method is as follows.
Actual value: 406.1273
Calculated value: 406.1267
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε66700), 268 nm (ε23000), 297 nm (ε12000)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3409, 3267, 2974, 2931, 1669, 1653, 1623, 1599, 1504, 1446, 1413, 1374, 1349, 1313, 1255, 1193, 1173, 1161, 1100, 1063, 1038, 998, 989 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.57 (3H, s), 2.60 (3H, s), 3.38 (2H, m), 4.40 (2H, s), 5.03 (1H , M), 6.22 (1H, br, s), 6.30 (1H, br, s), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.7 (t), 25.9 (q), 45.0 (t), 102.0 (d), 105.3 (s), 110 .2 (d), 113.1 (d), 122.7 (d), 127.3 (s), 129.3 (s), 132.5 (s), 133.3 (s), 145. 1 (s), 151.4 (s), 152.6 (s), 164.9 (s), 167.1 (s), 171.8 (s), 173.4 (s) ppm. Compound or salt thereof having the following physicochemical properties 1) Property of the substance: colorless acidic fat-soluble substance 2) Solubility: soluble in methanol, dimethyl sulfoxide, ethyl acetate 3) Molecular formula: C ₂₃ H ₂₃ NO ₈
4) Molecular weight: 441 (measured by TOF mass spectrum method)
5) The accurate mass [M + H] ⁺ measured by the high resolution TOF mass spectrum method is as follows.
Actual value: 442.1495
Calculated value: 442.1502
6) Ultraviolet absorption spectrum:
The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption:
213 nm (ε96200), 265 nm (ε41300), 297 nm (ε17500)
7) Infrared absorption spectrum:
The infrared absorption spectrum measured by the KBr pellet method shows the maximum absorption shown below.
3366, 3245, 2973, 2929, 2733, 1662, 1624, 1601, 1458, 1448, 1401, 1377, 1311, 1254, 1201, 1165, 1103, 1081, 1035, 995, 946 cm ⁻¹
8) ¹ H-nuclear magnetic resonance spectrum:
The ¹ H-nuclear magnetic resonance spectrum measured in deuterated methanol using the residual proton signal (3.31 ppm) as an internal standard is as follows.
1.50 (3H, s), 1.57 (3H, s), 2.61 (3H, s), 3.39 (2H, m), 4.38 (2H, s), 4.54 (2H , S), 5.04 (1H, m), 6.23 (1H, br, s), 6.30 (1H, br, s), 7.08 (1H, s) ppm
9) ¹³ C-nuclear magnetic resonance spectrum:
The ¹³ C-nuclear magnetic resonance spectrum measured in deuterated methanol using deuterated methanol (49.15 ppm) as an internal standard is as shown below.
17.9 (q), 24.7 (q), 24.8 (t), 25.9 (q), 45.1 (t), 50.3 (t), 102.0 (d), 105 .3 (s), 110.4 (d), 113.1 (d), 122.7 (d), 127.3 (s), 127.5 (s), 132.1 (s), 133. 3 (s), 145.1 (s), 151.4 (s), 152.4 (s), 164.9 (s), 167.1 (s), 170.8 (s), 171.8 (S), 172.8 ppm. The compound-producing bacterium according to any one of claims 1 to 5 belonging to the genus Stereum is cultured, and the compound according to any one of claims 1 to 5 is collected from the culture. The method for producing a compound according to any one of claims 1 to 5, wherein: Sterium sp. (Sterum sp.) SANK21205 strain (FERM BP-10465) or Sterium sp. (Sterum sp.) SANK21105 strain (FERM BP-10464) is cultured, and any one of claims 1 to 5 from the culture. The method for producing a compound according to any one of claims 1 to 5, wherein the compound according to claim 1 is collected. A microorganism belonging to the genus Stereoum and producing the compound according to any one of claims 1 to 5. Sterum sp. SANK21205 strain (FERM BP-10465). Sterium sp. SANK21105 strain (FERM BP-10464). A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 or a pharmacologically acceptable salt thereof as an active ingredient. The pharmaceutical composition according to claim 11, which is a therapeutic and / or prophylactic agent for at least one disease selected from the group consisting of diabetes, obesity, hyperlipidemia, hypertension and metabolic syndrome. An 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) inhibitor comprising the compound according to any one of claims 1 to 5 or a salt thereof.