JPH06192155A - Phenanthrene derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound exhibiting an interleukin-1 inhibitory activity and useful as an interleukin-1 inhibitor. CONSTITUTION:A compound of formula I (R<1> is a lower alkyl), formula II (R<2> is hydroxymethyl or carboxyl), formula III (R<3> is a lower alkoxycarbonyl or hydroxymethyl; R<4> and R<5> are each a lower alkoxy) or formula IV, e.g. 1,2,3,4,4a,5,8,9,10,10a-decahydro-1,4a-dimethyl-7-(1-methyl ester)-5,8-dioxo-1- phenanthrenecarboxylic acid. The compound of formula I can be synthesized by reacting a compound of formula V with diethyl phosphorocyanidate, triethyleneamine and proline methyl ester in a solvent (e.g. DMF) at 0 to 30 deg.C. Interleukin-1 is an important biological substance inducing and transferring systemic biological reactions against infection or inflammation and the compounds of formula I to IV are useful especially for improvement of chronic inflammation.

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 fa
ctor III (TRF-III), T-cell replacing factor M φ (TR
FM)], B cell activating factor (B-cel
l activating factor), B lymphocyte differentiation factor (B-cell d
The biologically active substances that have been reported under the names such as ifferentiation factor) are all interleukin-1 (IL-
1) It was decided to be unified into the name [Cellular
Immunol., 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 has a strong antitumor activity itself [Hirai Y. et al., "Gann Monograph on C
ancer Research ", Japan Scientific Societies Press,
Tokyo (1988)], at the same time, fever, increase in white blood cell count, activation of lymphocytes, induction of biosynthesis of acute phase protein in liver, etc.
It has been confirmed that it induces a reaction that occurs 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 joint synovium, 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- DR expression in the synovium of rheumato
id 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] At present, 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 pr
oduction in human peripheral blood mononuclearadhe
rent cells by a glucocorticoid hormone, J. Immuno
L., 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 5]

A compound represented by the formula [wherein R ¹ represents a lower alkyl group], the general formula (2):

[0011]

[Chemical 6]

A compound represented by the formula [wherein R ² represents a hydroxymethyl group or a carboxyl group], the general formula (3):

[0013]

[Chemical 7]

[Wherein R ³ represents a lower alkoxycarbonyl group or a hydroxymethyl group, and R ⁴ and R ⁵ represent a lower alkoxy group] and a compound represented by the formula (4)

[0015]

[Chemical 8]

It relates to a phenanthrene derivative selected from the group consisting of compounds represented by and salts thereof.

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. Examples of the lower alkoxy group include methoxy, ethoxy, propoxy, isoproxy, butoxy,
Examples thereof include linear or branched alkoxy groups having 1 to 6 carbon atoms such as tert-butoxy, pentyloxy and hexyloxy groups.

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

For example, the compound of the present invention represented by the above-mentioned general formula (2) is crozule (Tripterygium wilfordii Hook).
fil var. regelii Makino).

This extraction / isolation operation can be carried out in accordance with ordinary methods for extracting and isolating 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 adopt 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 a suitable solvent such as a 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 compound of the present invention can be purified and isolated.

The general formula (1a)

[0022]

[Chemical 9]

The compound of the present invention represented by the formula [wherein R ^{1 ′} represents a methyl group] is a compound (5) shown below, which is extracted and isolated by the same method as described above.

[0024]

[Chemical 10]

In addition, dimethylformamide (D
MF), diethyl ether, tetrahydrofuran (TH
F) and the like at a reaction temperature of 0 to 30 ° C., and diethylphosphorus cyanide, triethyleneamine, and proline methyl ester are 1 to 1.5 times mol, 2 to 5 times mol, and 1 to 1.5 times, respectively. It can be produced by reacting with about a double mole for a reaction time of 10 to 60 minutes.

Further, among the compounds of the general formula (1), the compound of the present invention in which R ¹ is a lower alkyl group other than a methyl group is
It can be produced by the same method as described above except that the corresponding proline alkyl stell is used.

Furthermore, among the compounds of the present invention represented by the general formula (3), the compound (3a) in which R ³ is a lower alkoxycarbonyl group is the general formula (2) obtained by the above extraction and isolation procedure. Compound (2a) in which R ² is a carboxyl group among these compounds is converted into a hydroxyl group by reducing the oxo group, and then the compound is reacted with an alkylating agent.

The above reduction reaction is carried out by catalytic reduction in the presence of a catalyst in a suitable solvent. As the solvent used, for example, water, acetic acid, alcohols such as methanol, ethanol and isopropyl alcohol, hydrocarbons such as hexane and cyclohexane, ethers such as diethylene glycol dimethyl ether, dioxane, tetrahydrofuran (THF) and diethyl ether, acetic acid. Examples thereof include esters such as ethyl and methyl acetate, polar solvents such as DMF, and mixed solvents such as these. The catalyst used is palladium, palladium-black, palladium-
Examples include carbon, platinum, platinum oxide, copper chromite, Raney nickel and the like. The amount of the catalyst used is usually about 0.02-1 times the amount of the compound (2a). The reaction is usually about -20 to 100 ° C, preferably about 0 to 70 ° C.
In the vicinity, at a hydrogen pressure of about 1-10 atm, 0.5-20
It takes about time.

The reduction reaction of compound (2a) can also be carried out by adding a 2 to 50-fold molar amount of an aqueous sodium hydrosulfite solution to compound (2a) in the solvent described in the above reduction reaction.

The subsequent alkylation reaction of the compound obtained by the above reduction reaction is carried out in a suitable solvent in the presence or absence of a basic compound. Here, as the alkylating agent, diazomethane, trimethylsilyldiazomethane, alkyl halides such as methyl iodide, FSO ₃ C
Lower alkyl sulfonic acid esters such as H ₃ , CF ₃ SO ₃ CH ₃ , (CH ₃ ) ₂ SO ₄ , (CH ₃ ) ₃ O ⁺ B
^{_{F 4 -, (C 2 H}} 5) 3 O + BF 4 - lower alkyl oxonium halide chelate salts such _{as, (C 2 H 5 O)} 3
Examples thereof include lower alkoxyoxonium halogenated chelate salts such as O ⁺ BF ₄ ⁻ . As the solvent used,
For example, water, alcohols such as methanol, ethanol, propanol, ethers such as dioxane, THF, diethyl ether, ethylene glycol monomethyl ether, halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, DMF, etc. Examples thereof include mixed solvents. Examples of the basic compound include carbonates such as sodium carbonate, potassium carbonate, sodium hydrogen carbonate and potassium hydrogen carbonate, metal hydroxides such as sodium hydroxide and potassium hydroxide. When diazomethane is used, the alkylating agent is usually used in a large excess amount, preferably about 10 to 20 times equivalent to the raw material compound, and the other alkylating agent is at least equimolar to the raw material compound. It is preferable to use an equimolar to 10-fold molar amount. The reaction is usually carried out at about -30 to 100 ° C, preferably at about -20 to 70 ° C, and generally the reaction is completed in about 0.5 to 20 hours.

Further, among the compounds of the present invention represented by the general formula (3), compounds in which R ³ is a hydroxymethyl group are
It can be produced by further reducing the compound of the general formula (3a) in which R ³ is a lower alkoxysulfonyl group. Usual reduction conditions can be adopted for the reduction reaction. For example a suitable solvent, LiAlH _4, diisobutylaluminum hydride (DIBAH) a reducing agent compounds such as (3a) with 1 to 10 mol per mol, at a reaction temperature of 0 to 30 ° C., the reaction time from 10 to 60 The target compound can be produced by reacting for about a minute. Examples of the solvent used here include THF and diethyl ether.

Further, the compound represented by the formula (4) can be produced by reducing the compound (5). The reaction condition in this case is that compound (5) is TH as a solvent.
Using an inert solvent such as F or dioxane, the reaction temperature is 0 to
At room temperature, 2 equivalents to an excess amount of a reducing agent such as sodium dithionite and potassium dithionite, and a reducing agent such as sodium bisulfite and bisulfite such as potassium bisulfite are used with respect to compound (5). Degree, reaction time 10 minutes ~
It can be produced by reacting for about 3 hours.

Among the phenanthrene derivatives of the present invention thus obtained, a compound having an acidic group, that is, a carboxyl group, can easily form a salt by reaction with a basic compound,
The present invention also includes such salts. 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. Examples of the solvent include water, lower alcohols such as methanol, ethanol and propanol, ethers such as dioxane and THF, and acetone, benzene, ethyl acetate, dimethyl sulfoxide, DMF, methylene chloride, chloroform and the like. . 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 is usually at least equivalent to the starting material, preferably 1
It is set to about 2 equivalents, and the desired salt can be obtained by the 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 phenanthrene derivative and its salt of the present invention have an IL-1 inhibitory activity and are useful as an IL-1 inhibitor. When the compound of the present invention is used for such pharmaceutical use, the compound may be used as it is or in the form of a general pharmaceutical preparation composition using a commonly used pharmaceutical preparation carrier.
It can be administered to humans and other animals. 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. The above pharmaceutical preparations include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, injections (solutions, suspensions, etc.) containing an effective amount of the compound of the present invention. Is mentioned. 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 formed by mixing the compound of the present invention as an active ingredient, and mixing this with an excipient such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic and the like. A capsule is prepared by mixing the above-mentioned active ingredient with an inactive formulation filler or diluent and filling the mixture into a hard gelatin capsule, a soft capsule 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 to be used in that case are water and physiological saline. Ordinary solubilizers, buffers, soothing agents and the like can be added to the injections and the like. Further, in the above-mentioned various forms of pharmaceutical preparations, colorants, preservatives, fragrances, flavors, sweeteners, etc., and other pharmaceuticals may 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 administered intravenously after being mixed with a normal fluid such as glucose and amino acid, or if necessary, it is intramuscularly, intradermally, subcutaneously or intraperitoneally administered alone. Further, the dose of the above-mentioned pharmaceutical preparation is appropriately selected according to the usage, the age of the patient, the sex and other conditions, the degree of disease and the like, but the amount of the compound of the present invention which is an active ingredient is usually about 0. The amount is preferably about 1 to 1000 mg, and the preparation can be administered in 1 to 4 divided doses per day. In the dosage unit form, the active ingredient is contained in an amount of about 1-60.
It is recommended to contain about 0 mg.

[0039]

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

[0040]

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 was 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 each of 10%, 20%, 30% methanol / ethyl acetate (v / v) and methanol, 500 ml each. Were collected to 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 liters of fraction (4-1)-(4)
-7) was obtained. Next, the above-mentioned fraction (4-4) (19
g) was subjected to silica gel column chromatography (Merck silica gel 70-230 mesh, 500 g), and
5%, 66%, 50% and 33% n-hexane / ethyl acetate (v / v%) and ethyl acetate were sequentially eluted with 2 l of each, and 100 ml of each eluate was collected, and fractions (4
(4-1) to (4-4-11) were obtained.

Further, the above fraction (4-4-6) 3.
5 g was subjected to silica gel column chromatography (Merck silica gel 70-230 mesh, 200 g), and 9
Eluted with 2 l of 5% chloroform / methanol (v / v%) and subjected to column chromatography twice using Sephadex LH-20 (500 ml, manufactured by Pharmacia).
Fractional elution with 0% methanol / chloroform and silica gel column chromatography (Merck silica gel 7
0-230 mesh, 100 g), fractionated and eluted with 1 l of 99% chloroform / methanol (v / v%) to purify,
This was recrystallized from methanol to give yellow needle-shaped crystals 1,
2,3,4,4a, 5,8,9,10,10a-decahydro-1,4a-dimethyl-7- (1-methylethyl)
0.057 g of -5,8-dioxo-1-phenanthrenecarboxylic acid was obtained.

Mp 207-209 ° C. [α] _D = + 80.5 (CHCl ₃ ; c1.0) IRν ^KBr _max cm ⁻¹ : 3400, 1730, 1696,
1650, 1470, 1270, 1240, 913, 8
00 UVλ ^MeOH _max nm (ε): 260 (15300) ¹ H-NMR (CDCl ₃ ) δppm: 1.01 (3H ×
2, d, J = 6.8 Hz, H-16, 17), 1.22
(3H, s, H-20), 1.31 (3H, s, H-1)
8), 1.0-1.2 (2H, m, H-1, H-3),
1.30 (1H, m, H-5), 1.55 (1H, br
d, J = 14.2 Hz, H-2), 1.79 (1H,
m, H-6), 1.96 (1H, m, H-2), 2.1.
5-2.29 (3H, m, H-3, H-6, H-7),
2.70 (1H, brd, J = 16.1Hz, H-
1), 2.75 (1H, dd, J = 19.5, 5.9H
z, H-7), 2.99 (1H, sept, J = 6.8.
Hz, H-15), 6.33 (1H, s, H-15) EI-MS m / z (rel.int.): 330 [M] ⁺ (3
5), 315 [M-CH ₃ ] ⁺ (15), 284 (5
5), 269 (100), 227 (44), 204 (3
9) HR-MS m / z: 330.1864 [M] ⁺ (C ₂₀ H ₂₆
O ₄ = 330.1831)

[0045]

[Example 2] 18.4 kg of black clover stem excluding the skin portion is shredded and extracted with 20 l of methanol at 60 ° C for 6 hours. Repeat this 3 times. The extract is concentrated under reduced pressure to give a crude extract (680 g). This crude extract was suspended in 2 liters of water, extracted with 2 liters of ethyl acetate three times, and the ethyl acetate layers were combined and concentrated under reduced pressure to obtain 140 g of an ethyl acetate extract.

140 g of the above ethyl acetate extract was subjected to silica gel column chromatography (Merck silica gel 70-
230 mesh, 1kg), 75%, 66%, 50
% And 33% n-hexane / ethyl acetate (v / v%) and ethyl acetate, each eluting sequentially with 3 l, then 95%
%, 90% and 80% ethyl acetate / methanol and 2 l each of methanol, and eluate 3 times each.
00 ml was collected to obtain fractions (1) to (20).

Of the above fractions, the fraction (1
The solvent of 0) was distilled off under reduced pressure, and 6.37 g of 6.5 g of the obtained residue was separated by Sephadex LH-20 (500 ml,
Column chromatography once using (Pharmacia), and fractionated and eluted with 2 l of methanol to obtain fractions (10-1) to (10-7).

Next, the above fraction (10-3) 1.2
4 g was subjected to silica gel column chromatography (Merck silica gel 70-230 mesh, 200 g), and 9
Elute with 1 liter each of 8% and 95% chloroform / methanol (v / v%) and similarly 98% chloroform / methanol (v / v%)
Elute with 1 liter and further high performance liquid chromatography (OD
S), eluted with 500 ml of 90% methanol / water (v / v%), and further subjected to silica gel column chromatography (Merck silica gel 70-230 mesh, 100 g).
Purified by eluting with 500 ml of 66% n-hexane / ethyl acetate to obtain 1,2,3,4,4a, amorphous powdery crystals.
24 mg of 5,8,9,10,10a-decahydro-1- (hydroxymethyl) -1,4a-dimethyl-7- (1-methylethyl) -5,8-dioxo-phenanthrene
Got

IRν ^KBr _max cm ⁻¹ : 3370,171
0, 1650, 1600, 1290, 1265, 108
0,906,755 UVλ ^MeOH _max nm (ε): 260 (11800) ¹ H-NMR (CDCl ₃ ) δppm: 1.04 (3H,
s, H-18), 1.09, 2.00 (each 3H, d, J
= 6.8 Hz, H-16,17), 1.24 (1H, b
rd, J = 11.2 Hz, H-5), 1.28 (3H,
s, H-20), 2.44 (1H, m, H-6), 1.
53 (1H, m, H-2), 1.68 (1H, m, H-
2), 1.80 (1H, brd, J = 13.6Hz, H
eq-3), 1.99 (1H, dd, J = 13.6,
6.8 Hz, Heq-6), 2.28 (1H, ddd,
J = 20.4, 11.6, 7.2 Hz, Hax-7),
2.69 (1H, dd, J = 20.4, 5.2Hz, H
eq-7), 2.75 (1H, brd, J = 13.2H)
z, Heq-1), 2.98 (1H, septd, J =
7.2, 1.0 Hz, H-15), 3.56, 3.78
(Each 1H, ABq, J = 11.2Hz, H-19),
6.32 (1H, d, J = 1.0 Hz, H-12) EI-MS m / z (rel.int.): 316 [M] ⁺ (8
5), 301 [M-CH _3] ⁺ (12), 298 [M-
H ₂ O] ⁺ (22), 285 (42), 283 [MC
H ₃ -H ₂ O] ⁺ (100), 241 (46), 203
(58), 91 (42), 43 (52) HR-MS m / z: 316.2073 [M] ⁺ (C ₂₀ H ₂₈
O ₃ = 316.2038)

[0050]

Example 3 3,4,4a, 5,8,9,10,10a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -obtained by the same extraction operation as in Example 1
5,8-Dioxo-2-phenanthrenecarboxylic acid 10
30.8 mg of triethylamine was added to 1.5 ml of a DMF solution in which 0 mg and 55 mg of proline methyl ester hydrochloride) were dissolved, and the mixture was stirred for 5 minutes. In addition, under ice cooling D
0.5 ml of a DMF solution containing 54.7 mg of EPC was added, followed by DM containing 92.4 mg of triethylamine.
0.5 ml of F solution was added, and the mixture was stirred at room temperature 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 anhydrous magnesium sulfate and concentrated. The resulting crude product is subjected to silica gel chromatography (eluate; chromatography: ethyl acetate = 1
0: 1), 3,4,4a, 5,8,9,1
0,10a-octahydro-1,4a-dimethyl-7-
(1-Methylethyl) -5,8-dioxo-2-phenanthrenecarboxamide-proline methyl ester 10
0 mg (74.7%) was obtained.

[0051]

Example 4 50 mg of the compound obtained in Example 1 at room temperature
Was dissolved in ₂ ml of THF, and the solution was added to 3 ml of water with Na ₂ S ₂
800 mg of O ₄ was added to the dissolved solution and stirred. After stirring for 3 hours, water was added to the reaction mixture, and the mixture was extracted with diethyl ether. The extract was washed with saturated brine and dried over anhydrous MgSO ₄ , then the solvent was evaporated to give a residue. Then, the residue was dissolved in 2 ml of acetone, 190 ml of K ₂ CO ₃ and 177 mg of methyl sulfate were added to the solution, and the mixture was heated under reflux for 2 hours. The reaction mixture was cooled to 0 ° C., 10% hydrochloric acid was added to neutralize the whole solution, and the mixture was extracted with diethyl ether. The extract was washed with saturated brine and dried over anhydrous MgSO ₄ , and the solvent was evaporated to give a residue.

The residue was subjected to silica gel column chromatography to obtain 95% n-hexane / ethyl acetate (v / v
%), Yellow needle crystals 1, 2, 3, 4, 4a,
9,10,10a-octahydro-5,8-dimethoxy-1,4a-dimethyl-7- (1-methylethyl) -1
-49 mg of phenanthrenecarboxylic acid methyl ester are obtained.

Mp 133-135 ° C. [α] _D = + 163.6 (MeOH; c0.26) IRν ^KBr _max cm ⁻¹ : 1719 ¹ H-NMR (200 MHz) (CDCl ₃ ) δ ppm:
1.12 (3H, s), 1.21 (3H × 2, d, J =
7.1 Hz), 1.27 (3H, s), 1.40-2.
66 (9H, m), 2.97-3.40 (3H, m),
3.65 (3H, s), 3.68 (3H, s), 3.7
6 (3H, s), 6.57 (1H, s) HR-MS m / z: 374.2437 [M] ⁺ (C ₂₃ H ₃₄
O ₄ = 374.2456)

[0054]

Example 5 38 mg of the compound obtained in Example 4 was dissolved in 0.5 ml of dry THF, and this solution was dried in THF.
To a suspension obtained by adding 30 mg of LiAlH ₄ to 0.5 ml was added dropwise at 0 ° C. After stirring for 30 minutes at room temperature, hydrous diethyl ether was carefully added at 0 ° C., and the reaction mixture was filtered using Celite. The filtrate is dried over MgSO ₄ ,
The solvent was distilled off to obtain an oily residue. Next, silica gel column chromatography (n-hexane-ethyl acetate,
9: 1 (v / v%)) to give colorless oils 1, 2,
3,4,4a, 9,10,10a-octahydro- (hydroxymethyl) -5,8-dimethoxy-1,4a-dimethyl-7- (1-methylethyl) -phenanthrene 3
1 mg was obtained.

[Α] _D = + 99.3 (MeOH; c0.
^{_{61) IRν KBr max cm -1:}} 3631 1 H-NMR (200MHz) (CDCl 3) δppm:
1.07 (3H, s), 1.21 (3H × 2, d, J =
6.8 Hz), 1.28 (3H, s), 1.34-2.
11 (8H, m), 2.49-3.40 (3H, m),
3.58 (2H, d, J = 10.7Hz), 3.66
(3H, s), 3.77 (3H, s), 3.84 (2
H, d, J = 10.7 Hz), 6.57 (1 H, s) HR-MS m / z: 346.2505 [M] ⁺ (C ₂₂ H ₃₄
O ₃ = 346.2507)

[0056]

Example 6 3,4,4a, 5,8,9,10,10a-octahydro-1,4a-dimethyl-7- (1-methylethyl) -obtained by the same extraction operation as in Example 1
5,8-Dioxo-2-phenanthrenecarboxylic acid 30
0 mg was dissolved in 10 ml of THF, and 16 ml of an aqueous solution in which 1.6 g of sodium dithionite was dissolved was added, and the mixture was stirred at room temperature for 3
Stir for 0 minutes. Water was added to the reaction mixture, extraction was performed with diethyl ether, the organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The crude product thus obtained was purified by silica gel column chromatography (eluent: chloroform: methanol = 100: 1),
4,4a, 9,10,10a-hexahydro-5,8-
290 mg of dihydroxy-1,4a-dimethyl-7- (1-methylethyl) -2-phenanthrenecarboxylic acid was obtained.

Mp 127 ° C. or higher (decomposition) ¹ H-NMR (DMSO-d ₆ ) δ ppm: 1.06 (3
H, s), 1.07 (3H, d, J = 6.7 Hz),
1.10 (3H, d, J = 6.7Hz), 1.21-
1.50 (2H, m), 1.97 (3H, s), 2.0
5-2.55 (5H, m), 2.81 (1H, dd, J
= 17.5, 4.4 Hz), 3.10-3.33 (2
H, m), 6.42 (1H, s), 7.18 (1H,
s), 8.47 (1H, s), 12.15 (1H, br)
s)

[0058]

[Pharmacological Test Example 1] RPMI-1640 medium (Nissui Pharmaceutical,
Penicillin 100 units / ml, streptomycin 0.1
μg / ml), 10% peripheral blood (heparin added), test compound and lipopolysaccharide (LPS,
3.3 μg / ml) was added, and the cells were cultured in an incubator containing 5% CO ₂ at 37 ° C. for 18 to 24 hours, and the culture supernatant was collected by centrifugation.

I released from cells by LPS stimulation
L-1α and IL-1β are measured by enzyme-linked immunosorbent assay (EI
The method A) was performed as follows. That is, a sample was added to and reacted with a 96-well plate for EIA on which a mouse monoclonal antibody against IL-1α or IL-1β was immobilized and subjected to blocking treatment. Wash after reaction, then IL-1α
Alternatively, a rabbit polyclonal antibody against IL-1β was added and further reacted. After washing the plate, horse radish peroxidase (PO
D)] Labeled anti-rabbit globulin was added and reacted. The unbound POD-labeled anti-rabbit globulin was washed off, a substrate solution (orthophenylenediamine and hydrogen peroxide) was added, and after the reaction, the absorbance at 492 nm was measured. IL-1
The release inhibition rate (%) was calculated by the following formula.

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 addition of the solvent. The absorbance at 492 nm when the culture supernatant obtained above was used as a sample is shown.

In this procedure, the results obtained when 1 × 10 ⁻⁵ g / ml of each compound obtained in the example was used as a test compound are shown in Table 1.

[0062]

[Table 1]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display location // A61K 31/12 AED 9283-4C 31/19 AEG 9283-4C 31/215 ABE 9283-4C 31 / 40 7431-4C (72) Inventor Kiyoto Goto 57 Myogami, Seto-cho, Naruto City, Tokushima Prefecture 57 (72) Inventor Yukihisa Ono 41-18 Kawakubo, Taihama, Kitajima-cho, Itano-gun, Tokushima Prefecture

Claims (1)

[Claims] 1. A general formula: [Wherein R ¹ represents a lower alkyl group], a compound of the general formula: [Wherein R ² represents a hydroxymethyl group or a carboxyl group], a compound represented by the general formula: [Wherein R ³ represents a lower alkoxycarbonyl group or a hydroxymethyl group, and R ⁴ and R ⁵ each represent a lower alkoxy group] and a compound represented by the formula: A phenanthrene derivative selected from the group consisting of compounds represented by and salts thereof.