JPH07110830B2 - Phenanthrene derivative - Google Patents

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel phenanthrene derivative, more specifically interleukin-1 (IL-1).
The present invention relates to the above-mentioned new derivative having the inhibitory activity of and useful as an IL-1 inhibitor.

[0002]

2. Description of the Related Art The phenanthrene derivative of the present invention is a novel compound which has not been published in the literature.

At the 2nd International Lymphokine Workshop, the Lymphocyte Activator
ting Factor; LAF), mitogenic protein
(Mitogenic Protein), Helper Peak-1 (Helper
peak -1), T lymphocyte replacement factor [T-cell replacing fac
tor III (TRF-III), T-cell replacing factor M φ (TRF
M)], B cell activating factor (B-cell
activating factor), B lymphocyte differentiation factor (B-cell di
fferentiation factor) and other physiologically active substances have been reported as interleukin 1 (IL-1).
It was decided to be unified into the following name [Cellular Imm
unol., 48, 433-436 (1979)]. This determination is based on the reason that the above-mentioned physiologically active substances are indistinguishable as substances and merely represent physiological activities by grasping them from different angles.

The above-mentioned IL-1 is known as an important biological substance that induces and transmits a systemic biological reaction to, for example, infection and inflammation, and also has a strong antitumor activity itself [Hirai Y. et al., "Gann Monograph on C
ancer Research ", Japan Scientific Societies Press,
Tokyo (1988)], but at the same time, fever, increase in white blood cell count, activation of lymphocytes, induction of acute phase protein biosynthesis in the liver, etc.
It has been found that it induces a reaction in the body during inflammation [Dinarello CA: Interleukin-1; Rev. Infec
t. Dis., 6, 51-95 (1984), Kluger, MJ, Oppenheim,
JJ & Powanda, MC The Physiologic, Metabolic
and Immunologic Actions of interleukin-1, Alan R.
Liss, lnc, New York (1985)].

The biological effects of IL-1 are diverse,
It is considered to be an important biological substance for maintaining homeostasis in the living body.
When an abnormality occurs in the L-1 production regulation function, IL-1 production is enhanced, and an excessive production is produced, it is considered to cause various diseases. For example, in rheumatoid arthritis, it has been reported that there is a strong correlation between the degree of inflammation of the synovial membrane, the degree of bone destruction, and the expression level of the HLA-DR antigen in the synovial tissue [Miyasaka, N., Sato. . K.,
Goto, M., Sasano, M., Natsuyama, M., Inoue, K. and
Nishioka, K., Augmented interleukin-1 production
and HLA- DRexpression in the synovium of rheumatoi
d arthritis patient Arthritis Rheum, 31, (4), 480-
486 (1988)]. Therefore, it is considered that various physiological actions of IL-1 can be blocked by inhibiting excessive IL-1 release from cells.

[0006] Currently, glucocorticoid hormone is used as a therapeutic agent for chronic inflammatory diseases, and it is known that a part of its action is suppression of IL-1 production [Lew, W., Oppenheim. , JJ & Matsushima, K., Anal
ysis of the suppression of IL- 1 α and IL- 1 β prod
auction in human peripheral blood mononuclear adher
ent cells by a glucocorticoid hormone, J. Immunol.,
140 (6), 1895-1902 (1988)], but the glucocorticoid has a disadvantage of causing various serious side effects due to its various physiological actions. Therefore, in the present field, there is no side effect as seen in glucocorticoids, and a new substance that is highly safe in terms of other toxicity and side effects and has high selectivity is desired especially in the therapeutic field of chronic inflammatory diseases. It is in.

[0007]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a new substance useful as an IL-1 inhibitor which meets the above-mentioned demands of the art.

That is, the present invention has the general formula (1)

[0009]

[Chemical 6]

[Wherein the group -A ... B- is a group

[0011]

[Chemical 7]

(R ¹ is a hydrogen atom or a lower alkyl group),
Basis

[0013]

[Chemical 8]

(R ² is a hydrogen atom , a formyl group or a lower alkanoyl group), a group

[0015]

[Chemical 9]

(R ² is the same as above) or a group

[0017]

[Chemical 10]

Is shown. ] Invitation to phenanthrene
Involved in conductors and their salts.

[0019]

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026]

[0027]

[0028]

[0029]

[0030]

In the present specification, examples of the lower alkyl group include methyl, ethyl, propyl, isopropyl,
Examples thereof include linear or branched alkyl groups having 1 to 6 carbon atoms such as butyl, tert-butyl, pentyl and hexyl groups. As also lower alkanoyl group, an isethionate <br/> Le, propionyl, butyryl, isobutyryl, pentanoyl, straight-chain or branched-chain alkanoyl group having 1 to 6 carbon atoms such as hexanoyl group can be exemplified, for example.

The lower alkanoyloxy group is, for example, a straight chain or branched chain alkanoyloxy having 2 to 6 carbon atoms such as acetyloxy, propionyloxy, butyryloxy, isobutyryloxy, pentanoyloxy and hexanoyloxy groups. A group can be illustrated.

As the lower alkoxy group, for example, a linear or branched alkoxy group having 1 to 6 carbon atoms such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentyloxy and hexyloxy groups. It can be illustrated.

The phenanthrene derivative of the present invention can be produced by various methods depending on its type.

For example, in the compound of the above general formula (1), the group-
Based on A ... B-

[0036]

[Chemical 11]

A compound of the same group -A ... B-

[0038]

[Chemical 12]

A compound of the same group --A ... B--

[0040]

[Chemical 13]

A compound of the same group -A ... B

[0042]

[Chemical 14]

Each of the compounds of
m wilfordii Hook fil var. regelii Makino).

This extraction / isolation operation can be carried out in accordance with ordinary extraction and isolation methods for general plant components. More specifically, the above operation is carried out by first extracting clozul using a normal polar solvent such as methanol or ethanol, concentrating the extract under reduced pressure to obtain a primary extract, and then the primary extraction. The method of collecting the target compound can be carried out based on various methods utilizing the physicochemical properties of the target compound from the product. As a method used for collecting the above-mentioned target compound, a usual method, for example, (1) a method utilizing a difference in solubility with impurities, (2) activated carbon, amberlite, silica gel, ion exchange resin, suitable of Sephadex, etc. It is possible to employ a combination of various methods such as a method of utilizing the difference in adsorption affinity for various adsorbents, (3) a method of utilizing the difference in distribution ratio between two liquid phases, and the like. As a more preferable collection method, for example, the primary extract is suspended in water, extracted with ethyl acetate, the extract is subjected to silica gel column chromatography, ethyl acetate-n-hexane mixed solution, Elute with an appropriate solvent such as methanol-chloroform mixed solution, and combine the obtained eluate with various column chromatography such as silica gel column chromatography, Sephadex LH20 column chromatography, gel filtration column chromatography, and high performance liquid chromatography. Each of them may be eluted with a suitable solvent such as an ethyl acetate-n-hexane mixed solution, a methanol-chloroform mixed solution, an acetone-chloroform mixed solution. Thus, the target compound of the present invention can be purified and isolated.

The compounds of the present invention other than those described above can be produced by subjecting the compounds of the present invention obtained from the above black melon to the raw materials and subjecting them to the following chemical treatments.

That is, in the compound of the above general formula (1), the group-
Based on A ... B-

[0047]

[Chemical 15]

And a compound in which R ¹ is a lower alkyl group (hereinafter referred to as “compound (1a)”) has the corresponding R ¹
It is produced by reacting a compound in which is a hydrogen atom (hereinafter referred to as "compound (1b)") with an alcohol in the presence of an acid catalyst in a suitable inert solvent. As the inert solvent used, various solvents that do not adversely affect the reaction, for example, ethers such as diethyl ether, tetrahydrofuran, dioxane, halogenated hydrocarbons such as dichloromethane, dichloroethane, chloroform, benzene, toluene, xylene, etc. Examples thereof include aromatic hydrocarbons, alcohols such as methanol, ethanol and propanol, and mixed solvents thereof. As the acid catalyst,
For example, aluminum chloride, stannic chloride, titanium tetrachloride,
In addition to boron trichloride, boron trifluoride-ethyl ether complex, Lewis acid such as zinc chloride, inorganic acid such as hydrochloric acid, nitric acid, sulfuric acid, organic acid such as trichloroacetic acid, trifluoroacetic acid, methanesulfonic acid, acetic acid, etc. An acid type ion exchange resin or the like can be used. In addition, as the alcohol to be reacted with the compound (1b) in the above, methanol, ethanol, propanol and the like can be used, and these also function as a solvent. The ratio of the alcohol used to the compound (1b) is usually at least equimolar amount, preferably about 1 to 30 times molar amount, and this reaction is generally carried out under the temperature condition of room temperature to the reflux temperature of the solvent, 1 to 72 hours, preferably 3
It takes about 24 hours.

Further, the compound (1a) can also be produced by the following diazotization method. That is, the method is a diazo compound corresponding to the ester residue in the esterified carboxyl group of the compound (1b) in the presence of an inert solvent,
For example, diazomethane, phenyldiazomethane, diphenyldiazomethane and the like are reacted, and examples of the inert solvent include halogenated hydrocarbons such as dichloromethane, dichloroethane, chloroform and carbon tetrachloride, dioxane, tetrahydrofuran, diethyl ether, dimethyl ether and the like. Ethers, nitro compounds such as nitromethane and nitrobenzene, alcohols such as methanol and ethanol, esters such as ethyl acetate and methyl acetate, aliphatic hydrocarbons such as hexane, heptane and octane, dimethylformamide, dimethyl sulfoxide, hexamethyl Examples include aprotic polar solvents such as phosphoric acid triamide, carbon disulfide, and the like. The ratio of the diazo compound used to the compound (1b) is at least an equimolar amount, preferably about 1 to 3 times the molar amount. The reaction proceeds well at -10 ° C to room temperature, and usually 10 minutes to 6 minutes.
The reaction ends in about time.

In the compound of the above general formula (1), the group -A
... based on B-

[0051]

[Chemical 16]

Wherein R ² is a formyl group or a lower alkanoyl group and the group —A ... B— is a group

[0053]

[Chemical 17]

[0054] a it is and R ² is a formyl group or a lower alkanoyl group, to the corresponding compounds wherein R ² is hydrogen atom, the formula ^{(8) (R 2 ')} 2 O or the general formula (9 R ^{2 ′} X [In the above formulas, R ^{2 ′} represents a formyl group or a lower alkanoyl group, and X represents a halogen atom. ] It manufactures by making the compound represented by these react.

This lower alkanoylation (including formylation
The reaction ) is carried out in the presence or absence of a basic compound. Examples of the basic compound used include alkali metal hydroxides, carbonates, bicarbonates, and organic bases such as pyridine and piperidine.
The reaction proceeds either with or without a solvent. Examples of the solvent include ketones such as acetone and methyl ethyl ketone, ethers such as diethyl ether and dioxane, aromatic hydrocarbons such as benzene, toluene and xylene, water and pyridine. The compound of the general formula (8) or (9) is used in at least an equimolar amount to the raw material compound, but it is generally preferable to use an equimolar amount to a large excess amount. Although the above reaction proceeds at 0 to 200 ° C, it is generally preferable to carry out the reaction at about 0 to 150 ° C. The reaction time of the reaction is generally about 0.5 to 20 hours.

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

Among the phenanthrene derivatives of the present invention thus obtained, a compound having an acidic group, that is, a phenolic hydroxyl group and / or a carboxyl group, can easily form a salt by a reaction with a basic compound, and the present invention is such. Also includes salt. Specific examples of the basic compound used in the production of the above-mentioned salt include alkali metal or alkaline earth such as sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogencarbonate and sodium hydride. Examples thereof include hydroxides, carbonates and hydrides of group metals. Further, for example, organic amines such as methylamine, ethylamine, isopropylamine, morpholine, piperazine, piperidine, and 3,4-dimethoxyphenethylamine can be used as the basic compound for salt formation.

The salt forming reaction with the above basic compound can be carried out in an appropriate solvent according to a usual salt forming reaction. As the solvent used, for example, water, methanol, ethanol, lower alcohols such as propanol, dioxane,
Other than ethers such as tetrahydrofuran, acetone,
Examples thereof include benzene, ethyl acetate, dimethyl sulfoxide, dimethylformamide, methylene chloride and chloroform. The reaction is usually carried out in the air or under oxygen-free conditions, preferably under an atmosphere of an inert gas such as nitrogen or argon at room temperature to about 100 ° C., preferably at room temperature to about 50 ° C. for about 5 minutes to 6 minutes. It will take about time. The amount of the basic compound used is not particularly limited, but it is usually equivalent or more, preferably about 1 to 2 equivalents relative to the starting material, and the desired salt can be obtained as described above.

The phenanthrene derivative of the present invention and its salt obtained by each of the above chemical treatment means can be isolated and purified by various usual separation means after the completion of each reaction. As the means, for example, solvent distillation, solvent extraction, precipitation, recrystallization, column chromatography, preparative chromatography and the like can be arbitrarily adopted.

The compounds of the present invention naturally include stereoisomers and optical isomers.

The thus obtained phenanthrene derivative of the present invention and its salt have IL-1 inhibitory activity and
-1 It is useful as an inhibitor.

The compound of the present invention can be administered to humans and other animals as it is or in the form of a general pharmaceutical preparation composition using a commonly used pharmaceutical preparation carrier.
The above-mentioned pharmaceutical preparation carrier, dosage form (dosage unit form), adjustment thereof, administration route thereof and the like can be the same as those of ordinary pharmaceutical preparations. That is, the above pharmaceutical preparations include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, which contain an effective amount of the compound of the present invention.
Examples include injections (solutions, suspensions, etc.). Each of the above-mentioned various forms of preparations may be prepared according to a conventional method, and the carrier used at that time may be any of various commonly used carriers. For example, tablets are prepared by using the compound of the present invention as an active ingredient, gelatin,
It is shaped by mixing with excipients such as starch, lactose, magnesium stearate, talc, and gum arabic. Capsules are prepared by mixing the above-mentioned active ingredient with an inert formulation filler or diluent and filling it into hard gelatin capsules, soft capsules or the like. Parenteral administration agents such as injections are produced by dissolving or suspending the compound of the present invention as an active ingredient in a sterilized liquid carrier, and preferred liquid carriers used in this case are water and physiological saline, which are prepared. Ordinary solubilizers, buffers, soothing agents and the like can be added to the injections and the like. Furthermore, in the above-mentioned various forms of pharmaceutical preparations, colorants, preservatives, fragrances, flavors, sweeteners, etc., and other pharmaceuticals can be contained, if necessary.

The amount of the phenanthrene derivative of the present invention and its salt to be contained as an active ingredient in the above-mentioned pharmaceutical preparation is not particularly limited and can be appropriately selected from a wide range, but it is usually 1 to 70% by weight in the whole pharmaceutical preparation composition. %, Preferably about 1 to 30% by weight. The administration method of the pharmaceutical preparation prepared above is not particularly limited, and for example, tablets, pills, powders, granules, capsules, etc. are orally administered, and injections (solutions, suspensions, etc.) alone or It is intravenously administered by mixing with a normal replenishing solution such as glucose or amino acid, or if necessary, it is intramuscularly, intradermally, subcutaneously or intraperitoneally administered alone. Furthermore, the dose of the above-mentioned pharmaceutical preparation is appropriately selected depending on the usage, the age of the patient, the sex and other conditions, the degree of disease, etc., but usually the amount of the compound of the present invention as an active ingredient is 1
The amount is preferably about 0.1 to 1000 mg per 1 kg of body weight per day, and the preparation can be administered in 1 to 4 divided doses per day. In addition, the active ingredient in the dosage unit form is about 1 to 6
It is recommended to contain about 00 mg.

[0070]

EXAMPLES In order to explain the present invention in more detail, a method for producing the phenanthrene derivative of the present invention will be given as an example, and then pharmacological test examples carried out on the compound of the present invention will be given.

[0071]

Example 1 108 kg of black clover stems are shredded and extracted with 200 l of methanol at room temperature for 7 days. The extract is concentrated under reduced pressure to give a crude extract. This crude extract is added to water 20
1 l of ethyl acetate and extracted with 20 l of ethyl acetate three times. The ethyl acetate layers were combined and concentrated under reduced pressure to obtain ethyl acetate extract 1
300 g are obtained.

1200 g of the above ethyl acetate extract was subjected to silica gel column chromatography (Merck silica gel 6
0,70-230 mesh, 1500g, manufactured by Merck)
20%, 40%, 60%, 80% ethyl acetate /
Sequentially elute with 10 l each of n-hexane (v / v) and ethyl acetate, and then sequentially elute with 10 l of 10%, 20%, 30% methanol / ethyl acetate (v / v) and methanol, 500 ml each. Collect and obtain fractions (1) to (11).

Of the above fractions, fraction (4)
The solvent was distilled off under reduced pressure, and 53 out of 57 g of the resulting residue was obtained.
g was subjected to silica gel chromatography (Merck silica gel 60, 70-230 mesh, 1200 g), and chloroform 10 l, 5% methanol / chloroform (v /
v) Fraction elution with 10 l, fraction (4-1) ~
(4-7) is obtained. Next, the above fraction (4-5,
6) are combined and concentrated under reduced pressure to give 19.96 g of residue. This residue was separated from Sephadex LH-20 (2000 m
column chromatography (Pharmacia Co., Ltd.) four times, fractionated and eluted with 3000 ml of methanol for purification, and recrystallized from methanol to give 3,4,4a, 5.
8,9,10,10a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-
2.654 g of 2-phenanthrenecarboxylic acid are obtained as pale yellow needles.

Molecular formula C ₂₀ H ₂₄ O ₄ (Mw328) [α] ²⁵ _D = + 296 ° (c = 0.14, chloroform) Rf ¹ : 0.36 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.16 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 2970, 1710, 16
80, 1650, 1605, 1298, 1266, 11
00,911,735 UV λ _max (CH ₃ OH) nm: 231 (ε = 1003)
0), 260 (ε = 15040) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.12 (6H, d, J = 6.8Hz), 1.18 (3
H, s), 1.14-1.56 (2H, m), 2.12
(3H, s), 2.22-2.26 (2H, m), 2.
39 (1H, ddd, J = 20.5, 11.2, 6.8
Hz), 2.42-2.63 (2H, m), 2.76-
2.82 (1H, m), 2.79 (1H, dd, J = 2)
0.5, 6.4 Hz), 3.01 (1H, septd, J =
6.8, 1.0 Hz), 6.38 (1H, d, J = 1.
0 Hz) ¹³ C-NMR (67.5 MHz, chloroform-d ₁ ).
δppm: 18.4 (q), 18.6 (t), 19.1 (q), 2
1.3 × 2 (q), 24.5 (t), 25.1 (t),
26.3 (d), 31.8 (t), 36.5 (s), 4
7.3 (d), 124.5 (s), 131.7 (d),
142.4 (s), 148.1 (s), 148.6
(S), 153.1 (s), 174.5 (s), 18
7.5 (s), 187.8 (s) EI-MS m / z (relative intensity): 328 [M] ⁺ (100), 313 [M-CH ₃ ]
⁺ (26), 310 (68), 295 (32), 282
(23), 267 (32), 229 (41), 204
(46), 191 (29), 189 (24) HR-MS m / z: 328.1662 [M] ⁺ , C ₂₀ H ₂₄ O ₄ calculated value 328.1675

[0075]

Example 2 55 g of 60 g of the residue obtained by distilling off the solvent of the fraction (5) obtained in Example 1 under reduced pressure was subjected to silica gel column chromatography (Merck silica gel 60,
70-230 mesh, 1000 g) and elute with 8 l of chloroform and 4 l of 5% methanol / chloroform (v / v) to obtain fractions (5-1 to 5-10). Fraction (5-1) was concentrated under reduced pressure to give a residue of 3.41 g.
To get This is subjected to silica gel column chromatography (Merck silica gel 60, 230-400 mesh, 3
00g) and 25% ethyl acetate / n-hexane (v /
v) 1 liter, 30% ethyl acetate / n-hexane (v / v) 50
0 ml, 50% ethyl acetate / n-hexane (v / v) 200 ml
Fractional elution with fractions (5-1-1 to 5-1-
7) is obtained. Fraction (5-1-3) was concentrated under reduced pressure to give 5,6,8,8a, 9,10-hexahydro-8-.
1.533 g of hydroxymethyl-4b, 8-dimethyl-2- (1-methylethyl) -1,4,7 (4bH) -phenanthrenetrione are obtained as a yellow-orange powder.

Molecular formula C ₂₀ H ₂₆ O ₄ (Mw330) [α] ²⁵ _D = + 336 ° (c = 0.21, chloroform) Rf ₁ : 0.61 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.19 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 3440, 1705, 16
50, 1465, 1300, 1235, 1040 UVλ _max (CH ₃ OH) nm: 258 (ε = 1402)
1) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.11 (6H, d, J = 6.8Hz), 1.28 (3
H, s), 1.35 (3H, s) 1.45 (1H, dd
dd, J = 13.7, 13.7, 13.7, 5.4H
z), 1.83 (1H, ddd, J = 13.7, 10.
3,5.9 Hz), 1.88 (1H, ddt, J = 1
3.2, 6.8, 2.0 Hz), 2.01 (1H, d
d, J = 13.2, 2.4 Hz), 2.29 (1H, d
dd, J = 20.0, 11.7, 6.8 Hz), 2.4
8 (1H, ddd, J = 15.6, 8.8, 5.9H
z), 2.69 (1H, ddd, J = 16.1, 10.
7, 5.9 Hz), 2.79-2.89 (2H, m),
3.00 (1H, septd, J = 6.8, 1.5Hz),
3.46, 4.05 (each1H, ABq, J = 11.2H
z), 6.37 (1H, s) ¹³ C-NMR (67.5 MHz, chloroform-d ₁ ).
δppm: 17.8 (t), 20.9 (q), 21.2 × 2
(Q), 22.3 (q), 25.4 (t), 26.3
(D), 34.1 (t), 34.3 (t), 37.0
(S), 50.4 (s), 51.6 (d), 65.3
(T), 131.7 (d), 142.4 (s), 14
7.8 (s), 153.2 (s), 187.2 (s),
187.4 (s), 220.0 (s) EI-MS m / z (relative intensity): 330 [M] ⁺ (100), 315 [M-CH ₃ ].
^{+ (16), 312 [M} -H 2 O] + (24), 300
(50), 299 (34), 285 (27), 269
(29), 229 (35), 215 (26), 175
(25) HR-MS m / z: 330.1854 [M] ⁺ , C ₂₀ H ₂₆ O ₄ calculated value 330.1831.

[0077]

[Example 3] The fraction (5-2) obtained in Example 2 was concentrated under reduced pressure to obtain 38.2 g of a residue. This is subjected to silica gel column chromatography (Merck silica gel 60, 1
000 g) and eluted with 4 l of chloroform, 3 l of 2% methanol / chloroform (v / v) and 3 l of 5% methanol / chloroform (v / v), and the fraction (5-
2-1 to 5-2-6) are obtained. Fraction (5-2
2) is concentrated under reduced pressure to obtain 3.278 g of residue. This was subjected to Sephadex LH-20 column chromatography (5
(00 ml) and eluted with 1 l of methanol to obtain fractions (5-2-2-1 to 5-2-2-3). Fraction (5-2-2-1) was concentrated under reduced pressure to give a residue of 0.91.
6 g was obtained, which was subjected to silica gel column chromatography (Merck silica gel 60, 230-400 mesh, 1
00g) and 2% methanol / chloroform (v / v)
Elute with 2 liters, then subject to silica gel column chromatography (Merck silica gel 60, 230-400 mesh, 300 g), 2% methanol / chloroform.
(v / v) Fractionated with 1 liter and purified, 1,2,3,4,4
a, 5,8,9,10,10a-decahydro-2-hydroxy-1- (hydroxymethyl) -1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-phenanthrene 97 mg Obtained as a crystalline substance.

Molecular formula C ₂₀ H ₂₈ O ₄ (Mw332) [α] ²⁵ _D = + 31.3 ° (c = 0.38, chloroform) Rf ₁ : 0.37 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.10 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 3369, 2971, 17
10, 1646, 1596, 1290, 1265, 10
80,906,755 UV λ _max (CH ₃ OH) nm: 260 (ε = 1184)
0) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.09 (3H, d, J = 6.8Hz), 1.10 (3
H, d, J = 6.8 Hz), 1.19 (1H, brd,
J = 11.7 Hz), 1.23 (3H, s), 1.25
(1H, m), 1.29 (3H, s), 1.38 (1
H, dddd, J = 19.0, 11.7, 11.7,
4.9 Hz), 1.81 (1H, ddd, J = 13.
7, 8.3, 3.9 Hz), 1.91-2.01 (2
H, m), 2.31 (1H, ddd, J = 20.5, 1
1.7, 7.3 Hz), 2.73 (1H, dd, J = 2)
0.5, 4.9 Hz), 2.81 (1H, ddd, J =
13.7, 3.9, 3.9 Hz), 2.98 (1H, se
ptd, J = 6.8, 1.0 Hz), 3.35 (1H,
d, J = 11.2 Hz), 3.48 (1H, dd, J =
11.7, 4.9 Hz), 4.26 (1H, d, J = 1)
1.2 Hz), 6.32 (1 H, d, J = 1.0 Hz) ¹³ C-NMR (67.5 MHz, chloroform-d ₁ ).
δppm: 17.4 (t), 20.9 (q), 21.3 × 2
(Q), 22.8 (q), 26.3 (d), 26.4
(T), 28.1 (t), 34.2 (t), 37.8.
(S), 43.1 (s), 51.8 (d), 64.0.
(T), 80.0 (d), 131.9 (d), 142.
7 (s), 149.6 (s), 153.1 (s), 18
7.7 (s), 187.8 (s) EI-MS m / z (relative intensity): 332 [M] ⁺ (19), 314 [M-H ₂ O] ⁺ (8
0), 299 (23), 296 [M-2H ₂ O] ⁺ (2
0), 281 (58), 203 (59), 91 (6)
6), 43 (100) HR-MS m / z: 332.1939 [M] ⁺ , C ₂₀ H ₂₈ O ₄ calculated value 332.1988.

[0079]

Example 4 Fraction (5-2) in Example 3
1), (5-2-2-2) and 3, in Example 1.
4,4a, 5,8,9,10,10a-octahydro-
1,4a-Dimethyl-7- (1-methylethyl) -5,
The crystal mother liquors of 8-dioxo-2-phenanthrenecarboxylic acid are combined and concentrated under reduced pressure to obtain 3.433 g of a residue. This was sequentially applied to Toyopearl HW-40F column chromatography (300 ml, Tosoh Corporation, 50% methanol / chloroform (v / v) 500 ml elution), Toyopearl HW.
-40F column chromatography (1800 ml, 50
% Methanol / chloroform (v / v) 2 l elution) and silica gel column chromatography (Merck silica gel 6
0,100 g, 33% ethyl acetate / n-hexane (v / v)
1 l elution), fractionated and purified to give 3,4,4a, 9,
10,10a-Hexahydro-8-hydroxy-1-
(Hydroxymethyl) -1,4a-dimethyl-7- (1
-Methylethyl) -2 (1H) -phenanthrenone 97
mg and crude 3b, 4,5,9b, 10,11-hexahydro-6-hydroxy-7- (1-hydroxy-1-methylethyl) -9b-methylphenanthro [1,2-C]
70 mg of furan-1 (3H) -one are each obtained as an amorphous substance.

Coarse 3b, 4, 5, 9b, 10, 11-
Hexahydro-6-hydroxy-7- (1-hydroxy-1-methylethyl) -9b-methylphenanthro [1,2-C] furan-1 (3H) -one was recrystallized from methanol to give colorless particles. 40 mg of a crystalline purified product is obtained.

3,4,4a, 9,10,10a-hexahydro-8-hydroxy-1- (hydroxymethyl)
-1,4a-Dimethyl-7- (1-methylethyl) -2
(1H) -phenanthrenone molecular formula C ₂₀ H ₂₈ O ₃ (Mw316) [α] ²⁵ _D = + 89.3 ° (c = 1.06, chloroform) Rf ₁ : 0.44 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.13 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ⁻¹ : 3412, 2962, 17
00, 1492, 1461, 1422, 1219, 10
38,757 UV λ _max (CH ₃ OH) nm: 217 (ε = 8620), 268 (ε = 2680), 3
40 (ε = 880) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.23 (3H, d, J = 6.8Hz), 1.25 (3
H, d, J = 6.8 Hz), 1.28 (3H, s),
1.34 (3H, s), 1.70 (1H, ddd, J =
13.2, 12.7, 12.7, 6.4 Hz), 1.9
8 (1H, m), 2.01 (1H, dd, J = 18.
1, 8.8 Hz), 2.10 (1H, dd, J = 13.
2,2.4 Hz), 2.48 (1H, ddd, J = 1
8.1, 7.8, 4.4 Hz), 2.57 (1H,
m), 2.63 (1H, dd, J = 15.1, 7.8H
z), 2.69 (1H, ddd, J = 15.1, 8.
8, 4.4 Hz), 2.92 (1H, dd, J = 16.
6,6.4 Hz), 3.12 (1H, sept, J = 6.8)
Hz), 3.54 (1H, d, J = 11.2Hz),
4.08 (1H, d, J = 11.2Hz), 6.80,
7.40 (each 1H, ABq, J = 8.3 Hz) EI-MS m / z (relative intensity): 316 [M] ⁺ (100), 301 [M-CH ₃ ]
⁺ (60), 285 (31), 283 (31), 271
(90), 241 (42), 199 (51), 147
(51) HR-MS m / z: 316.2006 [M] ⁺ , C ₂₀ H ₂₈ O ₃ calculated value 316.2038 3b, 4,5,9b, 10,11-hexahydro-6
- hydroxy-7- (1-hydroxy-1-methylethyl) -9B- methyl phenanthrylethynyl Ro [1,2-C] furan -1 (3H) - on molecular formula _{C 20 H 24 O 4 (Mw328} ) Rf 1: 0.50 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.18 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 3349, 3200, 29
33, 1729, 1671, 1569, 1401, 13
80, 1267, 1077, 1033, 755UVλ
_max (CH ₃ OH) nm: 220 (ε = 15840), 2
74 (ε = 1950), 283 (ε = 2040) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.03 (3H, s), 1.63 (3H, s),
1.69 (3H, s), 1.68 (1H, m), 1.88 (1H, dddd, J = 1
3.2, 13.2, 10.7, 7.3 Hz), 1.98
(1H, ddd, J = 13.2, 8.8, 3.6H
z), 2.39 (1H, ddd, J = 14.2, 6.
8,3.9 Hz), 2.48-2.52 (2H, m),
2.69 (1H, brd, J = 13.2Hz), 2.8
4 (1H, ddd, J = 18.6, 10.7, 8.8H
z), 2.98 (1H, dd, J = 18.6, 7.3H)
z), 4.77, 4.83 (each1H, ABq, 7.1H
z), 6.85 (1H, d, J = 8.3 Hz), 6.9.
6 (1H, d, J = 8.3 Hz), 9.20 (1H,
s) ¹³ C-NMR (67.5 MHz, chloroform-d ₁ ).
δppm: 18.2 (t), 19.7 (t), 22.3 (q), 2
2.5 (t), 30.3 (q), 30.4 (q), 3
2.6 (t), 36.3 (s), 41.0 (d), 7
0.6 (d), 76.1 (s), 114.8 (d), 1
22.6 (d), 123.3 (s), 125.0
(S), 127.8 (s), 145.9 (s), 15
3.6 (s), 163.2 (s), 174.3 (s) EI-MS m / z (relative intensity): 328 [M] ⁺ (3), 310 [M-H ₂ O] ⁺ ( 10
0), 295 (73), 277 (2), 185 (1
0), 147 (18), 115 (8) HR-MS m / z: 328.1629 [M] ⁺ , C ₂₀ H ₂₄ O ₄ calculated value 328.1675.

[0082]

Example 5 Fraction (5-2) in Example 3
6) is concentrated under reduced pressure to obtain 1.782 g of residue. This was subjected to silica gel column chromatography (Merck silica gel 60, 230-400 mesh, 400 g), and 5
Fractionated and purified with 1 liter of methanol / chloroform (v / v) to give 3,4,4a, 9,10,10a-hexahydro-
5-hydroxy-8-methoxy-1,4a-dimethyl-
0.124 g of 7- (1-methylethyl) -2-phenanthrenecarboxylic acid is obtained as an amorphous substance.

Molecular formula C ₂₁ H ₂₈ O ₄ (Mw344) [α] ²⁵ _D = + 171.2 ° (c = 1.0, chloroform) Rf ₁ : 0.19 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.16 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 3400, 2963, 26
20, 1681, 1620, 1412, 1262, 12
24, 1032, 798, 758 UV λ _max (CH ₃ OH) nm: 223 (ε = 946
0), 280 (ε = 2180), 287 (ε = 220)
0) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.17 (3H, d, J = 6.8Hz), 1.18 (3
H, s), 1.19 (3H, d, J = 6.8 Hz),
1.59 (1H, m), 1.64 (1H, m), 2.1
8 (3H, brs), 2.21 (1H, dd, J = 9.
2,6.4 Hz), 2.38 (1H, brd, J = 1
2.4 Hz), 2.41 (1 H, m), 2.63 (1
H, m), 2.68 (1H, m), 3.03 (1H, d
dd, J = 13.2, 7.6, 3.6 Hz), 3.10
(1H, dd, J = 16.8, 3.2Hz), 3.25
(1H, sept, J = 6.8Hz), 3.69 (3H,
s), 6.40 (1H, s) ¹³ C-NMR (100 MHz, chloroform-d ₁ ) δ.
ppm: 18.6 (q), 18.7 (q), 19.9 (t), 2
3.8 (q), 23.9 (q), 24.8 (t), 2
6.1 (d), 26.3 (t), 32.6 (t), 3
7.3 (s), 48.9 (d), 60.7 (q), 11
1.8 (d), 124.3 (s), 131.0 (s),
131.1 (s), 139.3 (s), 148.8
(S), 150.8 (s), 150.9 (s), 17
4.2 (s) EI-MS m / z (relative intensity): 344 [M] ⁺ (100), 329 [M-CH ₃ ]
⁺ (41), 311 (60), 283 (21), 245
(30), 241 (19), 205 (42) HR-MS m / z: 344.1963 [M] ⁺ , C ₂₁ H ₂₈ O ₄ calculated 344.1988.

[0084]

Example 6 3, 4, 4a, 5, 8, obtained in Example 1,
9,10,10a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-2-
63 mg of phenanthrenecarboxylic acid was dissolved in 8 ml of ether, and a diazomethane-ether solution was added under ice cooling to give 30
Stir for minutes. The ether was distilled off, and the obtained crude product was purified by silica gel column chromatography (eluent: ether / n-hexane = 1/2) to give 3,4,4.
a, 5,8,9,10,10a-octahydro-1,4
There was obtained 8.8 mg of a-dimethyl-7- (1-methylethyl) -5,8-dioxo-2-phenanthrenecarboxylic acid methyl ester.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: 1.10 (3H, d, J = 6.8Hz), 1.11 (3
H, d, J = 6.8 Hz), 1.17 (3H, s),
1.35 to 1.56 (2H, m), 2.00 (3H, m
d, J = 1.3 Hz), 2.15 to 2.55 (5H,
m), 2.71 to 2.83 (2H, m), 3.00 (1
H, septd, J = 6.8, 1.1 Hz), 3.73 (3
H, s), 6.36 (1H, d, J = 1.1Hz)

[0086]

[Embodiment 7] 5, 6, 8, 8a, 9, 1 obtained in Embodiment 2
0-hexahydro-8- (hydroxymethyl) -4b,
8-dimethyl-2- (1-methylethyl) -1,4,7
A mixture of 60 mg of (4bH) -phenanthrenetrione, 0.1 ml of acetic anhydride and 1 ml of pyridine was stirred at room temperature for 2.5 hours. The reaction mixture was poured into ice water and extracted with ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated. The obtained crude product was purified by silica gel column chromatography (eluent: ether / n-hexane = 1/2) to give 8- (acetoxymethyl) -5,6,8,8a, 9,10-hexahydro. -4
b, 8-dimethyl-2- (1-methylethyl) -1,
41 mg of 4,7 (4bH) -phenanthrenetrione was obtained.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: 1.07 (3H, d, J = 7.0Hz), 1.08 (3
H, d, J = 7.0 Hz), 1.41 (3H, s),
1.20 (3H, s), 1.77 (1H, dd, J = 1
2.7, 1.5 Hz), 1.88 to 2.03 (1H,
m), 2.00 (3H, s), 2.31 (1H, dd
d, J = 20.3, 11.3, 7.0 Hz), 2.48
(1H, ddd, J = 16.0, 6.6, 3.7H
z), 2.70-2.85 (2H, m), 2.91-
3.08 (2H, m), 4.05 (1H, d, J = 1
1.5Hz), 4.53 (1H, d, J = 11.5H
z), 6.35 (1H, d, J = 1.1 Hz)

[0088]

[Embodiment 8] 1, 2, 3, 4, 4a obtained in Embodiment 3,
5,8,9,10,10a-decahydro-2-hydroxy-1- (hydroxymethyl) -1,4a-dimethyl-
37- (1-methylethyl) -5,8-dioxo-phenanthrene 37 mg, acetic anhydride 0.1 ml and pyridine 0.5
The ml mixture was stirred at room temperature for 15 hours. The reaction mixture was poured into ice water and extracted with ether. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated. The obtained crude product was purified by silica gel column chromatography (eluent: ether / n-hexane = 1/3) to give 2- (acetyloxy) -1- (acetoxymethyl) -1,2,3,3. 4,4a, 5,8,9,10,10
27 mg of a-decahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxophenanthrene was obtained.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: 1.25 to 1.36 (1H, m), 1.50 to 1.68
(2H, m), 1.78 to 1.88 (2H, m), 2.
01 to 2.11 (1H, m), 2.07 (3H, s),
2.05 (3H, s), 2.26 (1H, ddd, J =
20.1, 11.8, 7.0 Hz), 2.70 to 2.8
8 (2H, m), 3.00 (1H, septd, J =
7.0, 1.1 Hz), 4.26 (1H, d, J = 1)
2.0Hz), 4.33 (1H, d, J = 12.0H
z), 4.57 to 4.55 (1H, m), 6.35 (1
H, d, J = 1.1Hz)

[0090]

[Embodiment 9] 3,4,4a, 5,8,9,10,10a
-Octahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-2-phenanthrenecarboxylic acid (150 mg) was dissolved in ethyl acetate (10 ml), and 10% palladium-carbon (15 mg) as a catalyst was dissolved in this solution. 3 ml of an ethyl acetate suspension of was added, the inside of the container was replaced with hydrogen gas, and the mixture was stirred at room temperature for 3 hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to give 3,4,4a, 9,10,10a-hexahydro-
5,8-dihydroxy-1,4a-dimethyl-7- (1
-Methylethyl) -2-phenanthrenecarboxylic acid 13
5 mg was obtained.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: 1.20 (3H, s), 1.23 (6H, d, J = 6.
8Hz), 1.55-1.76 (2H, m), 2.17
(3H, d, J = 1.1 Hz), 2.21-2.51
(3H, m), 2.53-2.70 (2H, m), 2.
88 (1H, dd, J = 6.7, 4.3Hz), 3.0
3-3.11 (1H, m), 3.07 (1H, sept, J
= 6.8 Hz), 6.41 (1H, s)

[0092]

[Embodiment 10] 3,4,4a, 5,8,9,10,10
600 mg of a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-2-phenanthrenecarboxylic acid was dissolved in 25 ml of tetrahydrofuran, and 6 g of sodium hydrosulfite was dissolved at room temperature. 50 ml of an aqueous solution was added and stirred for 30 minutes. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated. To this, 6 ml of pyridine and 0.6 ml of acetic anhydride were added, and the mixture was stirred at room temperature for 17 hours. Pour the reaction mixture into ice water,
It was extracted with ethyl acetate. The organic layer was washed with saturated saline,
The extract was dried over magnesium sulfate and concentrated. The obtained crude product is fractionated and purified by silica gel column chromatography (eluent: chloroform / methanol = 50: 1),
3,4,4a, 9,10,10a-hexahydro-5,
8-diacetyloxy-1,4a-dimethyl-7- (1
-Methylethyl) -2-phenanthrenecarboxylic acid (diacetyl form) 150 mg and 3,4,4a, 9,10,1
0a-hexahydro-5-acetyloxy-8-hydroxy-1,4a-dimethyl-7- (1-methylethyl)
-2-Phenanthrenecarboxylic acid (monoacetyl compound) 1
80 mg was obtained.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: diacetyl derivative 1.10 (3H, s), 1.15 (3H, d, J = 7.
0Hz), 1.18 (3H, d, J = 7.0Hz),
1.53-1.85 (2H, m), 2.16 (3H,
s), 2.15-2.26 (1H, m), 2.33 (3
H, s), 2.35 (3H, s), 2.31-2.73.
(6H, m), 2.75-2.95 (1H, m), 6.
78 (1H, s) monoacetyl body 1.13 (3H, s), 1.16 (6H, d, J = 7.
0Hz), 1.50-1.71 (2H, m), 2.15
(3H, s), 2.13-2.21 (1H, m), 2.
33 (3H, s), 2.31-2.46 (2H, m),
2.47-2.68 (2H, m), 2.75-2.93
(2H, m), 2.98-3.11 (1H, m), 6.
46 (1H, s)

[0094]

[Embodiment 11] 3,4,4a, 5,8,9,10,10
800 mg of a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-2-phenanthrenecarboxylic acid was dissolved in 30 ml of tetrahydrofuran, and 8 g of sodium hydrosulfite was dissolved at room temperature. 60 ml of an aqueous solution was added and stirred for 30 minutes. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and concentrated. This was dissolved in 3 ml of dimethylformamide, 1.2 ml of dimethylsulfate at room temperature, and then 930 sodium hydroxide.
0.7 ml of an aqueous solution in which mg was dissolved was added and stirred for 30 minutes. The reaction mixture was acidified with 1N hydrochloric acid and then extracted with ethyl acetate. The organic layer was washed with an aqueous solution of sodium hydrogen carbonate and then with saturated saline, dried over magnesium sulfate, and then concentrated. The obtained crude product was subjected to silica gel column chromatography (eluent: diethyl ether / n-
Hexane = 1:10) and purified to give 3,4,4a, 9,
10,10a-hexahydro-5,8-dimethoxy-
1,4a-Dimethyl-7- (1-methylethyl) -2-
720 mg of phenanthrenecarboxylic acid methyl ester was obtained.

¹ H-NMR (270 MHz, chloroform-d ₁ ) δ
ppm: 1.15 (3H, s), 1.21 (3H, d, J = 6.
8Hz), 1.23 (3H, d, J = 6.8Hz),
1.48-1.65 (2H, m), 2.05 (3H, q
-Like, J = 1.3 Hz), 2.11-2.23
(1H, m), 2.27-2.41 (2H, m), 2.
45-2.73 (2H, m), 2.97 (1H, dd
d, J = 13.3, 7.1, 3.3 Hz), 3.01-
3.13 (1H, m), 3.30 (1H, sept, J
= 6.8 Hz), 3.68 (3H, s), 3.75 (3
H, s), 3.80 (3H, s), 6.60 (1H,
s)

[0096]

Example 12 The fraction (4-4) obtained in Example 1 was concentrated under reduced pressure, and 19 g of the resulting residue was subjected to silica gel column chromatography (Merck silica gel 6).
0, 70-230 mesh, 1700 g, manufactured by Merck)
And eluted with 3 l of 20% ethyl acetate / n-hexane (v / v), and successively eluted with increasing ethyl acetate content to obtain fractions (4-4-1 to 4-4-11) .

Among the above fractions, the fraction (4-
4-3) was concentrated under reduced pressure to obtain 4.0 g of a residue. This was applied to Sephadex LH-20 (1500 ml, manufactured by Pharmacia) and 10% chloroform / methanol (v /
v) Fraction elution with 4 l, fraction (4-4-3-1
~ 4-4-3-8) was obtained.

The fraction (4-4-3-4) was concentrated under reduced pressure to obtain 0.41 g of residue. This is recrystallized from methanol to give 1,2,3,4,4a, 5,8,9,1
0,10a-decahydro-1,4a-dimethyl-7-
0.360 g of (1-methylethyl) -5,8-dioxo-1-phenanthrene aldehyde was obtained as pale yellow plate crystals.

Molecular formula C ₂₀ H ₂₆ O ₃ (Mw314) [α] ²⁵ _D = + 20.3 ° (c = 1.0, methanol) Rf ₁ : 0.78 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.64 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 2970, 1710, 16
50, 1600, 1297, 1231, 980, 91
5,754 UV λ _max (CH ₃ OH) nm: 260 (ε = 1457)
0) ¹ H-NMR (400 MHz, chloroform-d ₁ ) δ
ppm: 1.09 (3H, s), 1.10 (3H, d, J = 6.
8Hz), 1.11 (3H, d, J = 6.8Hz),
1.16 (3H, s), 0.97-1.13 (1H,
m), 1.03 to 1.18 (1H, m), 1.45 (1
H, dd, J = 13.2, 1.5 Hz), 1.57 (1
H, brd, J = 14.7 Hz), 1.68-1.80
(2H, m), 2.19-2.25 (2H, m), 2.
34 (1H, ddd, J = 20.0, 11.7, 6.8)
Hz), 2.72 (1H, brd, J = 13.2H
z), 2.80 (1H, ddd, J = 19.0, 5.
4, 1.0 Hz), 2.99 (1H, septd, J =
6.8, 1.0 Hz), 6.34 (1H, d, J = 1.
0 Hz), 9.77 (1 H, d, J = 1.0 Hz) ¹³ C-NMR (100 MHz, chloroform-d ₁ ) δ
ppm: 17.3 (t), 18.8 (t), 19.3 (q), 2
1.3 (q), 21.4 (q), 24.3 (q), 2
6.2 (t), 26.4 (d), 34.0 (t), 3
5.8 (t), 38.6 (s), 48.4 (s), 5
2.8 (d), 132.0 (d), 142.6 (s),
149.1 (s), 153.0 (s), 187.6
(S), 187.7 (s), 204.7 (s)

[0100]

Example 13 The solvent of the fraction (7) obtained in Example 1 was concentrated under reduced pressure, and 30 g of 200 g of the obtained residue was subjected to silica gel column chromatography (Merck silica gel 60, 70-230 mesh, 300 mesh).
g, manufactured by Merck & Co., Inc.) and eluted with 4 L of 5% methanol / chloroform (v / v) to give fractions (7-1 to 7-1).
3) was obtained.

Of the above fractions, the fraction (7-
7) was concentrated under reduced pressure to obtain 0.72 g of residue. This was applied to Sephadex LH-20 (300 ml, manufactured by Pharmacia) and fractionally eluted with 1 l of methanol to obtain fractions (7-7-1 to 7-7-7).

The fraction (7-7-3) was concentrated under reduced pressure to give a residue of 0.184 g. High-performance liquid chromatography (YMC PACKED COLUMN 343 I-15 ODS, 20 x
250 nm, manufactured by Yamamura Co., Ltd.), fractionated and eluted with 300 ml of methanol, and fractionated (7-7-3-1 to 7-7-3).
-3) was obtained. The fraction (7-7-3-2) was concentrated under reduced pressure to give a residue 0.03 g. High-performance liquid chromatography (TSK-GEL silica-60, 20 × 250n)
m, manufactured by TOSOH), fractionated and eluted with 100 ml of 5% methanol / chloroform (v / v), and the fraction (7
-7-3-2-1 to 7-7-3-2-3) was obtained. Fraction (7-7-3-2-3) was concentrated under reduced pressure to give 1,
2,3,4,4a, 5,8,9,10,10a-decahydro-1,4a-dimethyl-3-hydroxy-7- (1
10 mg of -methylethyl) -5,8-dioxo-1-phenanthrenecarboxylic acid were obtained as an amorphous substance.

Molecular formula C ₂₀ H ₂₆ O ₅ (Mw346) [α] ²⁵ _D = + 35.0 ° (c = 0.06, chloroform) Rf ₁ : 0.15 [5% methanol / chloroform (v /
v)] Rf ₂ : 0.04 [40% ethyl acetate / n-hexane (v
/ v)] IRν _max (KBr) cm ^-1 : 3430, 2960, 17
00, 1650, 1470, 1380, 1230, 10
30,910,760 ¹ H-NMR (400 MHz, C ₅ D ₅ N) δ ppm: 1.04 (3 H, d, J = 6.8 Hz), 1.06 (3
H, d, J = 6.8 Hz), 1.20-1.60 (3
H, m), 1.50 (3H, s), 1.66 (3H,
s), 2.05-2.16 (1H, m), 2.32 (1
H, dd, J = 19.5, 6.8 Hz), 2.38 (1
H, dd, J = 10.7, 4.4 Hz), 2.87 (1
H, dd, J = 19.0, 4.9 Hz), 3.03 (1
H, sept, J = 6.8 Hz), 3.16 (1H, d
d, J = 12.4, 2.9 Hz), 3.68 (1H, d
d, J = 13.7, 4.4 Hz), 4.98 (1H, t
t, J = 11.2, 4.4 Hz), 6.47 (1H,
s) ¹³ C-NMR (67.5 MHz, chloroform-d ₁ ).
δppm: 18.9 (q), 19.0 (t), 21.4 × 2
(Q), 26.4 (d), 26.6 (t), 28.7
(Q), 39.9 (s), 44.1 (s), 44.9.
(T), 45.7 (t), 52.6 (d), 64.4.
(D), 132.1 (d), 143.2 (s), 14
8.1 (s), 153.2 (s), 181.9 (s),
187.8 × 2 (s) EI-MS m / z (relative intensity): 346 [M] ⁺ (20), 328 [M-H ₂ O] ⁺ (1
00), 313 (50), 300 (48), 282 (8
7), 267 (67), 189 (33), 128 (3
7), 91 (53) The results of the pharmacological test of the compound of the present invention are shown below.

1. Interleukin-1 (IL-1) Release Inhibition Test from Human Peripheral Mononuclear Cells Peripheral blood of a healthy subject was collected, and mononuclear cells were collected using Ficoll-Pak (Pharmacia LKB Biotechnology Inc.) to obtain 100 units of penicillin. 2 ml per ml in RPMI-1640 medium (Nissui Pharmaceutical Co., Ltd.) containing 0.1 μg / ml, streptomycin 0.1 μg / ml, and 10% fetal bovine serum.
10 were suspended ^six cell mass. After adding 1 volume of the test compound to 1 volume of this cell suspension, lipopolysaccharide (LP
S) 1 volume of RPMI medium containing 10 μg / ml was added.
The cells were cultured in a humid air chamber containing 5% CO ₂ at 37 ° C for 24 hours, and the culture supernatant was collected by centrifugation.

IL-1α and IL-1β released from cells upon stimulation with LPS were measured by enzyme immunoassay (EI).
A) method was used. That is, a mouse monoclonal antibody against IL-1α or IL-1β is immobilized on a 96-well plate for EIA, and then blocking treatment is performed.
Samples are added to this plate and washed after the reaction, and then IL-1
A rabbit polyclonal antibody against α or IL-1β is added and further reacted. After washing the plate, horseradish peroxidase [horseradish peroxidase]
(POD)] Labeled anti-rabbit globulin is added and after reaction, unbound POD labeled anti-rabbit globulin is washed away. After reacting by adding a substrate solution (orthophenylenediamine and hydrogen peroxide), the absorbance at 492 nm is measured. IL
The -1 release inhibition rate (%) was calculated by the following formula.

IL-1 release inhibition rate (%) = 100 × (1−T ₄₉₂ ÷ C ₄₉₂ ) where T ₄₉₂ is the absorbance at 492 nm when the culture supernatant containing the test compound was used as a sample, and C ₄₉₂ was The absorbance at 492 nm is shown when a culture supernatant containing a solvent is used as a sample.

In this procedure, when the compound of the present invention (3 × 10 ^-6 g / ml) obtained in Example 1 was used as the test compound, I
The inhibition rates of L-1α and IL-1β release were 25% and 49%, respectively.

When the compound of the present invention obtained in Example 2 (3 × 10 ^-6 g / ml) was used as a test compound, IL-1
α and IL-1β release inhibition rates are 78% and 94%, respectively
Met.

2. Rat adjuvant arthritis test 8.
Five-week-old female Wistar-Lewis rats were used.
0.5 mg of Mycobacterium butyricum (Mycobact
erium butyricum) killed bacteria (manufactured by Difco) was suspended in 0.1 ml of liquid paraffin as an adjuvant,
It was sensitized in the skin of the rat ridge. The sensitization day was regarded as the 0th day, and the arthritis observed in the hind limbs thereafter was measured using a foot volume meter (MK-500 manufactured by Muromachi Kiki Co., Ltd.).

The drug was suspended in a 5% gum arabic solution and orally administered to rats on a schedule of 5 times a week from the day of adjuvant sensitization.

As a result, a significant arthritis inhibitory effect was observed in the 50 mg / kg-administered group of the present invention obtained in Example 1 (Table 1).

[0112]

[Table 1]

In Table 1 above, the numerical values show the average paw volume (ml) and standard error of the left and right hind limbs on the 16th day. The numbers in parentheses indicate the number of animals used in each group. ** means significant difference (p <in Tukey)
0.01) is shown.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 31/19 9455-4C 31/215 ABG 9455-4C 31/22 ABE 9455-4C 31/34 ( 72) Inventor Toshiko Kuwahara 21 Nakase Nakanoketsu, Nakashigo, Matsushige-machi, Itano-gun, Tokushima 21 (72) Inventor Masaaki Takai 62-14 Minamikawa, Matsushige-machi, Itano-gun, Tokushima Prefecture (72) Inventor Yukihisa Ono 41-18 Kawakubo, Taihama, Kitashima-cho, Itano-gun, Tokushima Prefecture (72) Inventor Yoshihiro Taniguchi 5-2 Kanaoka, Kawauchi-cho, Tokushima-shi, Tokushima 407 La Foret Kawauchi 407 (72) Inventor Sachiko Manabe Kaga, Kawauchi-cho, Tokushima-shi, Tokushima Prefecture Suno 463-10 (72) Inventor Takahiro Asakuni 81 Nyu Doi, Ouchi Town, Okawa District, Kagawa Prefecture

Claims (1)

[Claims] 1. A general formula: [In the formula, the group -A ... B- is a group (R ¹ is a hydrogen atom or a lower alkyl group), a group: (R ² is a hydrogen atom , a formyl group or a lower alkanoyl group), a group: (R ² is the same as above) or a group Indicates. Fenansure <br/> emissions derivatives and salts thereof, characterized in that represented by].