JPS6317884A - Tricyclo compound - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (R is amine group derived from carbamic acid) or its salt. EXAMPLE:17-Allyl-12-[ 2-{ 4-(3-carboxypropylcarbamoyloxy )-3-methox-cyclohexyl }-1-methylvinyl ]-1,14-dihydroxy-23, 25-dimethoxy-1 3, 19,21,27-tetramethyl-11, 28-dioxa-4-azatricyclo[ 22,3,1,0 <4.9> ]octacos-18-ene-2,3,10,16-tetraone. USE:Immunosuppressive agent and antibacterial agent. PREPARATION:An acyl group derived from carbamic acid such as carboxymethylcarbamoyl is introduced into the FR-900506 substance of formula II.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to pharmacologically active tricyclo compounds or salts thereof.

More specifically, the present invention relates to a novel tricyclo compound or a salt thereof having pharmacological activities such as immunosuppressive activity and antibacterial activity.

That is, an object of the present invention is to provide a novel tricyclo compound or a salt thereof useful for the treatment of transplant rejection, graft-versus-host disease caused by bone marrow transplantation, autoimmune diseases, infectious diseases, etc., and for pre-fatization.

In the treatment of rejection reactions and autoimmune diseases that occur during transplants such as heart transplants and liver transplants, the immunosuppressants used are the most important factor influencing the therapeutic outcome.

Azathioprine and cyclosporin A are known as existing immunosuppressants, but the immunosuppressive effects of these compounds are not satisfactory, and various side effects have also been reported. The development of an inhibitor was desired.

The inventors of the present invention conducted various studies in order to develop such immunosuppressants, and as a result, they found that several compounds such as FR-900506, which is produced by bacteria belonging to the genus Streptomyces, have extremely strong immunosuppressive effects. find out that it has
He succeeded in elucidating the chemical structures of these compounds and synthesizing their derivatives, and filed a patent application (Japanese Patent Application Laid-Open No. 148181-1981).

After further intensive research, they succeeded in synthesizing a new tricyclo compound derivative and completed the present invention.

The novel tricyclo compound of this invention can be represented by the following formula.

(In the formula, R means an acyl group derived from carbamic acid.) In the tricyclo compound (I) of this invention, optical isomers and geometric isomers derived from asymmetric carbon atoms and double bonds, etc. One or more pairs of stereoisomers may exist, and such combomers or isomers are also included within the scope of this invention.

As the salt in tricyclo compound (1), non-toxic,
Conventional pharmaceutically acceptable salts, such as alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, ammonium salts such as triethylamine salts, N-benzyl-N
- Salts with inorganic or organic bases such as amine salts such as methylamine salts.

The tricyclo compound (I) of this invention is produced by the following production method.

Production method I FR-900506 substance [wherein R has the same meaning as before, R1 means a protected carboxy(lower) alkylcarbamoyl group, and R2 means a carboxy(lower)alkylcarbamoyl group, respectively. The definitions used in the M book are detailed below.

'Lower grade and naru words are next subbuko 1 to 6 unless otherwise specified.
means individual.

Examples of the acyl group derived from carbamic acid in R include lower alkylcarbamoyl groups having one or more suitable substituents such as carboxy or protected carboxy, and specific examples thereof include, for example, Carboxy(lower)alkylcarbamoyl groups such as carboxymethylcarbamoyl, carboxyethylcarbamoylbutylcarbamoylbamoyl, carboxyhexylcarbamoyl, e.g. trimethylsilylmethoxycarbonylethylcarbamoyl, trimethylsilylethoxycarbonylpropylcarbamoyl, triethylsilylethoxyluponylpropylrubamoyl, tertiary butyl Protected carbokynes (such as tri(lower)alkylsilyl(lower)alfkidicarbonyl(lower)alkylcarbamoyl groups such as dimethylsilylethoxyl, luponylprobyl, rubamoyl, trimethylsilylbroboxy, luponylbutyl, rubamoyl, etc.
(lower) alkylcarbamoyl groups, and more preferred are carboxy(C1804)alkylcarbamoyl groups and tri(C1-C4)alkylsilyl(C1804)alkylcarbamoyl groups.
The most preferred examples of the 1-C4) alkylcarbamoyl group include a 3-carboxypropylcarbamoyl group and a 3-(2-trimethylsilylethoxycarbonyl)propylcarbamoyl group.

1 protected carboxyl (lower) alkyl carbamoyl group in R1, and 1 carboxy (lower) alkylcar/hemoyl group in R2.

Examples of the acyl group include those exemplified in 1 above for the acyl group derived from carbamic acid.

The method for producing the tricyclo compound (1) of the present invention will be explained below.

Production method 1 Tricyclo compound (1) and its salt are prepared using FR-900
It can be obtained by introducing a device group consisting of one carbamine into the 506 substance.

The FR-900506 substance used as an ingredient is, for example, Streptomyces tsukubaensis (5tre to
m ces tsukubaensis ) No,
Streptomyces like 9993 (821 molars 1 ming)
It was isolated in pure form from a fermentation culture of the R-900506 substance-producing strain belonging to the genus. This Streptomyces tsukubaensis No. 9993 was newly removed from a soil sample collected in Toyosato Town, Tsukuba District, Ibaraki Prefecture, and its freeze-dried specimen was sent to the National Institute of Microbial Technology, Agency of Industrial Science and Technology (Ibaraki Prefecture). 1-1-3 Higashi, Yatabe-cho, Tsukuba-gun, on October 5, 1984, with deposit number 788.
It was deposited as No. 6, and then switched to the Budapest Treaty route of the same depositary on October 19, 1985.
It has been deposited under the new deposit number 927.

Examples of the agent for introducing the acyl group derived from carbamic acid include isocyanate compounds, and more preferably lower alkyl isocyanates having a protected carboxyne group.

This reaction is usually carried out in a solvent such as benzene that does not adversely affect the reaction. Although the reaction temperature is not particularly limited, it is preferably carried out under cooling or heating.

Production method 2 Compound (I-b) and its salt can be obtained by subjecting compound (1-a) to a carboxy-protecting group elimination reaction.

The elimination reaction of the carboxy protecting group is carried out by a conventional method,
When the protected carboxy group is tri(lower)alkylsilyl(lower)alkoxycarbonyl, react with tetra(lower)alkylammonium fluoride such as tetrabutylammonium buttide, potassium fluoride, or hydrogen fluoride. This is done by

This reaction is usually carried out in a solvent such as tetrahydrofuran which does not adversely affect the reaction. Although the reaction temperature is not particularly limited, it is usually carried out under cooling or heating.

Tricyclo compounds obtained by production methods 1 and 2 (
1) can be isolated and purified by conventional methods such as extraction, precipitation, fractional crystallization, recrystallization, and chromatography.

During the reaction of the production method 1 and 2 of the surface or the post-treatment of the reaction mixture, the Hnhomer and the stereoisomer due to the asymmetric carbon depth or double bond of the starting compound and the target compound are converted to other Hnhomer and stereoisomers. , and such cases are also included within the scope of the present invention.

The tricyclo compound (I) of this invention has an immunosuppressive effect,
It has pharmacological effects such as antibacterial effects, and is therefore effective against rejection reactions to organ or tissue transplants such as the heart, kidney, liver, bone marrow, and skin, graft-versus-host reactions caused by bone marrow transplants, rheumatoid arthritis, and the whole body. It is useful for the treatment and prevention of autoimmune diseases such as sexual lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, and uveitis, and infectious diseases caused by pathogenic bacteria.

The tricyclo compound (I) can be administered, for example, in the form of solid, semi-solid or liquid pharmaceutical preparations, mixed with organic or inorganic carriers or excipients suitable for external, internal or parenteral administration. can. This pharmaceutical preparation consists of tablets,
pellets, capsules, suppositories, liquids, emulsions,
They can be used as suspensions and other suitable forms, eg, mixed with conventional non-toxic, pharmaceutically acceptable carriers. Carriers that can be used include water, gluten, lactose,
Acacia comb, gelatin, mannitol, starch paste, magnesium trisilicate, talc, cornstarch, keratin, colloidal silica, potato starch, urea, and other carriers suitable for producing solid, semisolid or liquid preparations. In addition, excipients, stabilizers, thickeners, colorants, and fragrances may be used. The target compound is contained in the pharmaceutical preparation in an amount sufficient to exert the desired effect depending on the course or condition of the disease.

When this pharmaceutical preparation is used for humans, it is preferably administered parenterally or internally.

The therapeutically effective amount of trinculo compound (I) varies depending on the age and severity of the disease of the individual patient being treated, but usually
Approximately 0.01 to 1000 mg of active ingredient per mouthful, preferably 0.
．． 1 to 500 mg, more preferably 0.5 to 1100
ff1 is used in the treatment of diseases, generally at an average dose of about 0.
5mg, lmg, 5mg, 10mg,
50mg, 100mg, 250mg.

500 mg is administered.

The present invention will be described below with reference to Examples.

Production Example Isolation of Streptomyces tsukubaensis No. 9993 Streptomyces
Tsukubaensis No. 9993 was isolated.

Approximately 1 gram of soil collected in Toyosato Town, Tsukuba District, Ibaraki Prefecture was put into a sterile test tube and... Boost the JR bacteria water to a capacity of 5-.
Mix the mixture with a tube buzzer for 10 seconds and let stand for 10 minutes. The supernatant is serially diluted 100 times with sterile water. The diluted solution (Q, 1 mf) was added to Suzapetsk agar containing thiamine hydrochloride (
Sucrose 30g, [6 sodium 3&, dipotassium phosphate 1g, magnesium sulfate 0.5g, potassium chloride 0.5g, ferrous sulfate 0.01g, thiamine hydrochloride 0.1
g, agar 20g, tap water 10100O: pH 7.2)
Spread on a Petri dish containing. Incubate for 21 days at 30°C, transfer the colonies grown on the plate to a slant medium [yeast-malt extract agar (ISP medium 2) and incubate at 30°C for 10 days. Mrs. Tsukubaensis No. 9993
I found out.

Late M Glycerin (1%), corn starch (1%), glucose (0,5%), cottonseed flour (1%), corn starch liquor (0,5%), dried yeast (0,5%) and A preculture medium (1 oomQ) containing calcium carbonate (0.2%) at pH 6.5 is placed in a 500-Erlenmeyer Fusco and sterilized at 120° C. for 30 minutes. One platinum loop of a slant culture of Streptomyces tsukubaensis No. 9993 is inoculated into the medium and cultured at 30''C for 4 days.

The resulting culture is transferred to preculture medium (20 liters) in a 30 liter jar fermenter that has been previously sterilized at 120"C for 30 minutes. The culture is incubated at 30"C for 2 days to remove the preculture medium. 16 liters of soluble starch (4.5%), cornstarch strength (1
%), dried yeast (1%), calcium carbonate (0.1%)
, Adekanol (antifoaming agent, trade name, manufactured by Asahi Denka) (0,1
%) at pH 6.8 (1600 liters) and cultured at 30°C for 4 days.

Isolation and Purification The culture broth thus obtained was grown on a diatom (2
5 kg). Dip the mycelium mass into acetone (
500 liters) to obtain 500 liters of extract liquid. This acetone extract and filtrate (1350 liters) were combined, and the nonionic adsorption resin 1 Diaion HP20
+ (trade name, manufactured by Mitsubishi Kasei Corporation) (100 liters) column. Washing with water (300 liters) and 50% aqueous acetone (300 liters) is followed by elution with 75% aqueous acetone. Evaporate the eluate under reduced pressure and reduce the remaining volume to 3
00 liters. This is extracted three times with ethyl acetate (20 liters). The ethyl acetate extract was condensed under reduced pressure to obtain an oily residue. This oily residue is mixed with twice the weight of acidic silica gel (special silica gel, grade 12, manufactured by Fuji Davison) and the mixture is slurried in ethyl acetate. The solvent is evaporated and the resulting dry powder is subjected to column chromatography with n-hexane packed on the same acidic silica gel (8 liters) as above. The column was developed with n-hexane (30 liters), n-hexane-ethyl acetate mixture (4:1 v/v, 30 liters), and ethyl acetate (30 liters). Fractions containing the target compound are collected and concentrated under reduced pressure to obtain an oily residue. This oily residue is mixed with twice the weight of acidic silica gel and the mixture is slurried in ethyl acetate. After distilling off the solvent, the resulting powder was
'＼Resubmit the chromatography using a column of acidic silica gel (3.5 liters) packed with xane. ) and developed with ethyl acetate (10 liters). Fractions containing the target compound are collected and concentrated under reduced pressure to obtain a yellow oil. This oily residue was washed with a mixture of n-hexane and ethyl acetate (1:1).
Silica gel (230-400 mesh, manufactured by Merck & Co., Ltd.) (v/v, 300 m ) and packed in the same solvent system (
Chromatography using a 297 Torr column was performed. n-hexane-ethyl acetate mixture (1:1 v/v, 1
0 liters: 1:2 v/v, 6 liters) and ethyl acetate (6 liters).

The fractions containing the first target compound were collected and concentrated under reduced pressure at °C, F.
The R-900506 material is obtained as a white powder (34 g). This white powder is dissolved in acetonitrile and concentrated under reduced pressure. The concentrated solution was left at 5°C overnight to form prismatic crystals (22
, 7g). Purified FR-90 by recrystallization in the same solvent
0506 substance (13.6 g) is obtained as colorless prismatic crystals.

Infrared absorption spectrum 2830, 1?45.1720.1700.1645°1450, 1380.1350.1330.1310°1285, 1170.1135.1090.1050°1030.1000.990.960 (sh).

918 cm-' Example FR-900506 substance (310 mg), 2-trimethylsilylethyl 4-isocyanatobutyrate (350
mg) and triethylamine (6 drops) in anhydrous benzene (4 mQ) are stirred at 50°C for 2 hours. The reaction mixture, left overnight at room temperature, is concentrated to dryness under reduced pressure. The resulting residue was subjected to silica gel chromatography using chloroform to obtain 17-allyl-12-[2-[3-
Methoxy-4-(3-(2-trimethylsilylethoxycarbonyl)propylcarbamoyloxy)cyclohexyl-1-methylvinylco-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-
Tetramethyl-11,28-dioxa-4-azatricyclo[zz,a,t,o”cooctacos-18-ene-
2,3,10,16-titraone (99 mg) is obtained.

This product was dissolved in tetra(n-butyl)ammonium fluoride (0,12 mfi) in tetrahydrofuran (2,5 mfi).
mole) for 20 minutes at room temperature. The reaction mixture was heated under reduced pressure at t! After condensation to dryness, the resulting residue was purified by silica gel preparative chromatography (developing solvent: chloroform-methanol, 5:1 v/v) to obtain 17-allyl-12-[2-(4-(3-carboxylic) propylcarbamoyloxy)-3-methoxycyclohexyl)-1-methylvinyl]-1,14-dihydroxy-23,25-dimethoxy-13,19,21,
27-tetramethyl-11,28-dioxa-4-azatricyclo[22,3,1,0'°9 Cooctafus-1
8-ene-2゜3.10.16-titraone (ts, t
mg).

Claims (1)

[Claims] 1) A tricyclo compound or a salt thereof represented by the general formula ▲ Numerical formula, chemical formula, table, etc. ▼ (I) (in the formula, R means an acyl group derived from carbamic acid). 2) Claim 1 in which R is carboxy or a protected carboxy (lower) alkyl carbamoyl group
) Compounds described in section 2. 3) 17-allyl-12-[2-{4-(3-carboxypropylcarbamoyloxy)-3-methoxycyclohexyl)-1-methylvinyl]-1,14-dihydroxy-23,25-dimethoxy-13,19 ,21,2
Claims that are 7-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^4^,^9]octacos-18-ene-2,3,10,16-tetraone The compound described in item 2). 4) 17-allyl-12-[2-[3-methoxy-4-
(3-(2-trimethylsilylethoxycarbonyl)propylcarbamoyloxy)cyclohexyl]-1-methylvinyl]-1,14-dihydroxy-23,25-
dimethoxy-13,19,21,27-tetramethyl-
11,28-dioxa-4-azatricyclo[22.3
．． 1.0^4^,^9] Octacos-18-en-2,
The compound according to claim 2), which is 3,10,16-tetraone.