WO1998040393A1

WO1998040393A1 - Novel mycaminosyltylonolide derivatives

Info

Publication number: WO1998040393A1
Application number: PCT/JP1998/000954
Authority: WO
Inventors: Tomio Takeuchi; Sumio Umezawa; Osamu Tsuchiya; Toshiaki Miyake
Original assignee: Microbial Chemistry Research Foundation
Priority date: 1997-03-10
Filing date: 1998-03-09
Publication date: 1998-09-17
Also published as: JP4106104B2; JPH10251291A; AU6121098A

Abstract

Novel mycaminosyltylonolide derivatives represented by general formula (I) or salts thereof, and drugs comprising the same, wherein X represents a hydroxyl group or a hydrogen atom; R represents -CH2SR1 (wherein R1 represents an alkyl, cycloalkyl, or aryl group); and Me represents a methyl group. The above compounds have excellent antimicrobial activities against Gram-positive and Gram-negative bacteria and are useful as macrolide antibiotics.

Description

Description New myosinyltylonolide derivative Technical field

The present invention relates to a novel myosinyltylonolide derivative having antibacterial activity. More specifically, the present invention relates to a novel myosinyltyronolide or 4'-deoxymycinosyltyronolide having antibacterial activity, which is a 3- (1-alkyl (alkyl- or cycloalkyl- or phenylthiomethyl) ) For derivatives. Background technology

Tylosin is a macrolide antibiotic produced by Streptomyces flagell NRRL 022 and 273, and has a broad antibacterial spectrum against gram-positive and gram-negative bacteria. (JP-B-36-226649). Thai mouth synth has a mycaminose attached to the 5th position of a lactone ring called Tylonolide, and a mycinose attached to the 23rd position of the lactone ring. It has a structure having mycarose bonded to the 4-position hydroxyl group of the mycosis moiety (H. Umezawa, Recent Advances in Chemistry and Biochemistrv of Antibiotics, 964, pp. 62-63, and Letters, j No. 34, pp. Pp. 2339-2345 (1964) .Thai mouth sin also inhibits spirochetes, protozoa, etc., and reduces parasitic parasites such as chickens in poultry. It is used as a feed additive because it has inhibitory activity and promotes livestock growth.

The formula for Thai mouth is as follows (A)

(Wherein, Me represents a methyl group, and the same applies to the following). Hydrolysis of Thai Mouth Shin under mildly acidic conditions yields My Power Loin and the following formula (B)

In generating a Desumaikoshin (Desmycosin) represented it is known _(and, as a tie opening Shin derivatives, the following formula (C)

It is known that mycaminosyltylonolide represented by, and 4′-deoxy and 3,4′-dideoxy derivatives of mycaminosyltylonolide have antibacterial activity. (Showa 57-280100). It is also known that a 3-deoxy derivative of myricinosyltylonolide has an antibacterial activity (Japanese Patent Application Laid-Open No. 2-275894).

Many compounds have been known in which various substituents have been introduced into the hydroxyl group at position 23 of the macrolactone ring of myricinosyltylonolide (Macrolide Antibiotics p85-125 (1984) Academic press, inc). Japanese Patent Application Laid-Open No. 5-199296 discloses an MT derivative in which the 23-position is substituted with a methylthiomethyloxy group.

On the other hand, the hydroxyl group at the 3-position of the macrolactone ring has low reactivity and it is difficult to introduce a substituent, and it has not been considered that the introduction of the substituent can enhance the antibacterial activity. . Actually, as an MT derivative having a substituent introduced at the 3-position of the macrolactone ring, acetyl-O— (Bull. Chem. Soc. Jpn., 54 (12), 3837-3845 (1981)) Compounds substituted by T-carbonyl-O (J. Antibiot., 39 (12), 1724-1735 (1986)) are known. Have been. Japanese Patent Application Laid-Open No. 62-50909 discloses a cladinosyl group as a 3-position substituent. In addition, compounds in which {TH} (tetrahydroxypyranyloxo group) and the like are substituted at the 23-position and the 3-position are known (Japanese Patent Application Laid-Open No. 5500/1999). These antibacterial activities are insufficient. there were.

There is always a need to provide pharmaceuticals with excellent antibacterial and biological activities or new substances that are useful as veterinary drugs. In particular, there is a need to provide a new and useful derivative of Thai oral synth, which has been widely used as a feed additive or animal drug, by chemical synthesis. There is also a need to provide new derivatives of myricinosyltyronolide. Disclosure of the invention

The present inventors have carried out various studies for the purpose of synthesizing new myominosyltylonolide derivatives. As part of these studies, the present inventors have developed an amino acid methyl group, a cycloalkylthiomethyl group, or an arylthiomethyl group at the 3-position hydroxyl group of myosinyl tynonolide ゃ 4'-dexoximeinylsyltyronolide. Research was conducted to introduce a group.

As a result of the research, starting from my-minosyltylonolide by the method described below, a new compound, 3- (1-alkyl (monoalkyl or cycloalkyl), Mono- or arylthiomethyl) derivatives. Also, starting from 4'-doxime-minosyltylonolide, synthesizing 3- (1- (alkyl- or cycloalkyl-alkyl- or arylthio-methyl) derivatives of 4'-dioxime-minosyltylonolide I also succeeded.

The present inventors have succeeded in newly synthesizing myo-minosyltylonolide (hereinafter sometimes abbreviated simply as Τ）) or 4'-deoxymi-nosyltyronolide (hereinafter simply referred to as “4”). (Also abbreviated as DMT) 3- 3- (alkyl- or cycloalkyl- or arylthiomethyl) derivatives have excellent antibacterial activity against certain Gram-positive and Gram-negative bacteria Thus, the derivatives are recognized as being useful as fungicides, and their derivatives are expected to have an activity of inhibiting the growth of spirochetes and protozoa and other biological activities.

Therefore, in the present invention, the following general formula (I) H

[Wherein X represents a hydroxyl group or a hydrogen atom, R represents a group CHSR ¹ (where R ¹ is an alkyl group, a cycloalkyl group or an aryl group), and Me represents a methyl group]. Provided is a 3〇- (alkyl- or cycloalkyl- or phenylthiomethyl) derivative of myosinyltylonolide or 4'-deoxymycinosyltylonolide, or a salt thereof.

The myominosyltylonolide derivatives of the general formula (I) according to the present invention can be two groups of derivatives represented by the following formulas (Ia) and (Ib), respectively.

(1) The following equation (Ia)

(Wherein R and Me are as defined above), a 3-O 1 (alkyl-, cycloalkyl-, or arylthiomethyl) derivative of myosinosyltyronolide, ie, 3-O — (Alkyl or cycloalkyl mono or arylthiomethyl) myosinosyltylonolide.

(2) The following equation (Ib)

(Where R and Me are the same as above). 3-D- (alkyl- or cycloalkyl- or arylthiomethyl) derivatives of ride, ie, 3_〇_ (alkyl- or cycloalkyl- or arylthiomethyl) -1 4'-Deoxime-force minosyltylonolide .

The 3-O-substituted monomycinosyltyronolide derivative or the 3- (1-monosubstituted 1-4'-deoxymiminosyltylonolide derivative of the general formula (I) according to the present invention is introduced. In the substituent R, that is, in the group CH ₂ SR ¹ , when R ¹ is an alkyl group, the alkyl group is a straight-chain or branched (〇, to 〇,.) Alkyl group, preferably ( C ^ CJ alkyl group, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, n-heptyl group and n-hexyl group When R ¹ is a cycloalkyl group, the cycloalkyl group is a (C ₃ -C ₁₀ ) cycloalkyl group, preferably a (C ₄ -C ₆ ) cycloalkyl group, for example, a cyclopropyl group, Cycloheptyl group and cycle It can be a mouth hexyl group or the like.

The Ariru group, phenyl group, 0 ₆ ～〇, ... such as a naphthyl group And a phenyl group.

Preferable specific examples of the 3-O-substituent—CH ₂ SR ¹ of the derivative of the general formula (I) according to the present invention include a methylthiomethyl group, an ethylthiomethyl group, an n-propylmethyl group, and an n-hexylthio group. A methyl group, a cyclohexylthiomethyl group and a phenylthiomethyl group.

Preferred specific examples of the derivatives of the general formula (I) according to the present invention include the following compounds.

(1) 3-O-methylthiomethyl myominosyltyronolide [compound (la-1)]

(2) 3-O-ethylthiomethyl MT [Compound (Ia-2)]

(3) 3_0—n-hexylthiomethyl MT [Compound (Ia-3)]

(4) 3-dicyclohexylthiomethyl MT [Compound (Ia-4)]

(5) 3-O-phenylthiomethyl MT [Compound (Ia-5)]

(6) 3-D-methylthiomethyl _ 4'-doximeminosyltylonolide [Compound (Ib-1)] (7) 3-Perethylthiomethyl-14'-DMT [Compound (Ib_2)]

(8) 3-〇-n-hexylthiomethyl-4'-DMT [Compound (Ib-3)]

(9) 3-P-cyclohexylthiomethyl-14'-DMT [Compound (Ib-4)]

(10) 3_〇-phenylthiomethyl-14'-DMT [Compound (Ib_5)]. In the above, MT is an abbreviation for My Power Minosyltyronolide, and 4 '— D

MT is an abbreviation for 4'-deoxy maiminosyltyronolide.

(Manufacturing method)

The method for producing the derivative of the general formula (I), that is, 3-O-substituted mono-aminosyltylonolide or 3-O-substituted 4'-deoxydimycinosyl-minosyltylonolide according to the present invention will be described. The derivative of the general formula (I) can be produced by a six-step method comprising the following steps (a) to (f).

Step (a): When the myo-minosyltylonolide of the above formula (C) is dissolved in methanol containing hydrogen chloride and the solution is left at room temperature for 30 minutes, the aldehyde group at the 20-position is converted to dimethyl acetal. The protected myosinyltylonolide 20-dimethyl acetal is produced (see JP-A-57-281100). The corresponding 20-dimethyl acetal is produced by using 4'-deoxymycinosyltylonolide instead of myosynyltylononolide.

The general formula (I I I) thus generated

(Where X is a hydroxyl group or a hydrogen atom) When the reaction solution containing MT 20-dimethylacetal or 4'-DMT 20_dimethylacetal is distilled under reduced pressure to dryness, a solid residue is obtained. Thus, MT or 4'-DMT20_dimethyl acetal of the above formula (III) can be prepared.

In the step (a), the compound represented by the formula (III): MT or 4′-DMT 20-dimethyl Selectively protect only the 2'- and 4'-hydroxyl groups (when the raw material is MT) or only the 2'-hydroxyl group (when the raw material is 4'-DMT) of lacetal with an acetyl group (abbreviated as Ac) To this end, the compound of formula (III) is reacted with acetic anhydride in acetonitrile anhydride at room temperature. By this reaction, the following formula (IV)

(Wherein Ac is an acetyl group and X ′ is an acetyloxy group or a hydrogen atom). 2 ′, 4′-Diacetylacetyl MT 20-dimethylacetal or 2′-O-acetyl- 4'-DMT 20—Dimethyl acetal is formed.

The reaction solution containing the compound of the formula (IV) is concentrated under reduced pressure. The obtained residue is dissolved in toluene, washed with a saturated aqueous solution of sodium hydrogencarbonate, dehydrated, and concentrated to dryness under reduced pressure to obtain a compound of the formula (IV) as a solid.

Step (b): In order to selectively protect the hydroxyl group at position 23 of the compound of formula (IV) with a t-butyldimethylsilyl group (abbreviated as TBS), the compound of formula (IV) is treated with anhydrous dimethylformamide ( Reaction with t-butyldimethylsilyl chloride in the presence of imidazole in DMF) at room temperature. By this reaction, the following formula (V)

(In the formula, Ac and X 'are the same as described above, and TBS is a t-butyldimethylsilyl group.) 2', 4'-diacetylacetyl- 23_0-t-butyldimethylsilyl MT 20-dimethylacetal or 2'-0-acetyl-23-t-butyldimethylsilyl-1 4'-DMT 20-dimethylacetal is produced. The reaction solution is concentrated, the residue is dissolved in toluene, and the compound of the formula (V) is purified by silica gel high-performance column chromatography to obtain the compound of the formula (V) as a solid.

Step (c): In order to introduce a substituent of the formula CH ₂ SR 1 (where R ¹ has the aforementioned meaning) into the hydroxyl group at the 3-position of the compound of the formula (V), the compound of the formula (V) At a temperature of or near 50 ° C in a mixed solution of acetic anhydride and acetic acid.

° ιι (V I)

〇

(Wherein, R ¹ is an alkyl group, a cycloalkyl group, or a phenyl group)

The alkyl or phenyl methyl sulfoxide is reacted in an amount of about 40 to 50 molar equivalents. This gives the following equation (V I I)

(Wherein, Ac, X ', TBS and R ¹ are the same) 2 represented by', 4 '- di - O-Asechiru one 23- O-t-heptyl dimethylsilyl one 3- O-(alkyl 1- or cycloalkyl- or phenylthiomethyl) MT 20-dimethylacetal or 2'-O-acetyl-23-0-t-butyldimethylsilyl-13-O- (alkyl or cycloalkyl- or phenylalkyl) (Dithiomethyl) -1 4'-DMT 20-dimethyl acetal is produced.

The reaction solution containing the compound of the formula (V 1 I) is concentrated under reduced pressure. When the obtained syrup-like substance is purified by silica gel single-pressure ram chromatography, the compound of the formula (VII) is obtained as a solid.

Step (d): In order to remove the O-acetyl group from the compound of the formula (V II), the compound of the formula (V

II) The compound is heated in anhydrous methanol at a temperature of 50 ° C. or higher. As a result, a deacetylation reaction occurs, and the following formula (VIII)

(In the formula, X represents a hydroxyl group or a hydrogen atom, and R ¹ and TBS are the same as described above.) 23-〇-t-butyldimethylsilyl — 3 —〇_ (alkyl or cycloalkyl mono or phenyl) (Ruthiomethyl) MT20_dimethylacetal or 23-〇-tert-butyldimethylsilyl-13-〇 (alkyl- or cycloalkyl- or phenylthiomethyl) -1'-DMT20-dimethylacetal is produced. Expression (V

When the reaction solution containing the compound of III) is concentrated, the compound of formula (VIII) is obtained as a residue.

Step (e): Treatment of the compound of formula (VIII) with tetrabutylammonium fluoride in tetrahydrofuran (THF) at room temperature to remove the TBS group at position 23 from the compound of formula (VIII) I do. This gives the following equation (I X)

(Wherein X and R ¹ are the same as described above), and 3-O— (alkyl mono- or cycloalkyl mono- or phenylthiomethyl) MT or 14′-DMT 20-dimethyl acetal is produced.

The reaction solution containing the compound of the formula (IX) is concentrated under reduced pressure. The resulting syrupy substance is purified by silica gel-one column chromatography to isolate the compound of the formula (IX).

Step (f): In order to remove the dimethyl acetal group at the 20-position as a protecting group for the aldehyde group from the compound of the formula (IX), the compound of the formula (IX) is treated with a low concentration of sulfuric acid in 1,4-dioxane. Room temperature in the presence of hydrochloric acid or a low concentration of hydrochloric acid in acetonitrile And hydrolyze. As a result, an MT derivative or a 4′-DMT derivative of the general formula (I), which is a target product, is produced.

The reaction solution containing the compound of the formula (I) is made slightly alkaline with an aqueous solution of sodium hydrogen carbonate, and then extracted with chloroform. The port-mouth form extract is dehydrated and further concentrated under reduced pressure. The obtained residue is purified by silica gel single column chromatography (eluent: chloroform-form-methanol-28% aqueous ammonia, eluted with 15: 1: 0.1) to give a compound of the formula (I).

The compound of the present invention thus produced can be isolated and purified as a salt, a hydrate, a solvate, or a crystalline polymorph of the compound as free. The salt of the compound (I) of the present invention can also be produced by subjecting the salt to a conventional salt formation reaction.

The compound of the present invention forms a salt. Specifically, it is an acid addition salt with an inorganic acid or an organic acid, and a pharmaceutically acceptable salt is preferable. Specific examples of these salts include mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid and phosphoric acid, or formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, With organic acids such as fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or toluenesulfonic acid, or acidic amino acids such as aspartic acid or glutamic acid Addition salts can be mentioned.

The starting compounds of the above-mentioned production methods include novel substances, but they can be produced by applying the production methods described in the examples, methods based on the production methods, or modified methods arbitrarily practicable by those skilled in the art. Isolation and purification are performed by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various types of chromatography. Industrial applicability

Compounds of specific examples of the derivatives of the general formula (I) according to the invention were tested for their antibacterial activity.

For this test, the minimum inhibitory concentrations (MIC, mcg / ml) of these representative compounds against various bacteria were determined by a standard multiple dilution method. The medium used was a nutrient agar medium, and MIC was evaluated after culturing at 37 ° C for 18 hours. The results obtained are summarized in Tables 1 and 2 below. For comparison, the same applies for myominosyltyronolide (abbreviation: MT) and 4'-deoxy myominosyltyronolide (abbreviation: 4'-1 DMT). The measured MIC (mcg / mI) is also shown in Table 1 or Table 2 below. table 1

, 1 set, test compound M I Cu g / ml)

^ Test strain la- 2 la-4 la-5 MT

Staphylococcus aureus 193 0.39 <0.2 <0.2 1.56

Staphylococcus aureus FDA209P <0.2 <0.2 <0.2 0.78

Staphylococcus aureus MS9861 0.39 0.39 0.39 1.56

Staphylococcus aureus MSI 0225 0.78 0.39 0.39 1.56

Staphylococcus aureus Smith 0.39 0.78 0.39 0.78

Micrococcus luteus PCIIOOI <0.2 0.39 <0.2 0.39

Bacillus subtilis NRRL B-558 0.78 0.39 0.39 3.12

Corynebacterium bovis 1810 1.56 0.39 0.39 3.12

Shigella dysenteriae JSH910 0.78 1.56 0.78 0.78

Salmonella enteritidis I89l 3.12 3.12 1.56 3.12

(Test compound M I C g / ml)

Test bacteria ~~ .lb-1 lb-4 lb-5 4'-DMT

Staphylococcus aureus 193 0.39 <0.2 <0.2 0.78

Staphylococcus aureus FDA209P 0.39 <0.2 <0.2 0.39

Staphylococcus aureus MS9861 0.78 1.56 0.39 0.78

Staphylococcus aureus MSI 0225 0.78 0.78 0.39 0.78

Staphylococcus aureus Smith 0.78 0.39 0.39 0.39

Micrococcus luteus PCIIOOI 0.2 0.39 0.2 0.78

Bacillus subtilis NRRL B-558 0.78 0.39 0.39 1.56

Corynebacterium bovis 1810 0.78 <0.2 0.39 0.78

Escherichia coli K-12 LA290 R55 12.5 25 6.25 12.5

Shigella dysenteriae JSH910 6.25 1.56 12.5 6.25 It was confirmed that the compound of the present invention exhibited excellent antibacterial activity against Gram-positive bacteria and Gram-negative bacteria.

As a result, the compounds of the present invention are useful as fungicides, and their derivatives are expected to have activity to inhibit the growth of protozoa and other biological activities.

Preparations containing one or more of the compound of the present invention or a salt thereof as an active ingredient are prepared using carriers, excipients and other additives usually used for preparation of preparations.

Administration may be oral, such as tablets, pills, capsules, granules, powders, or liquids, or parenteral, such as intravenous or intramuscular injections, suppositories, or transdermals. Good No. The dose should be appropriately determined according to the individual case in consideration of the symptoms, age of the subject, gender, etc., and usually about 0.01 to 1 OOOmg / day, preferably 0 to 1 mg / day for an adult in the case of oral administration. 0.1 to 100 mg / day, for parenteral administration, 0.1 to 100 mg / day for adults, preferably about 0.01 to 5 mg / day. ~ 4 divided doses.

As the solid composition for oral administration according to the present invention, tablets, powders, granules and the like are used. In such a solid composition, the one or more active substances include at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone. , Meta-silicic acid, mixed with magnesium aluminate. In accordance with the usual practice, the composition may contain additives other than inert diluents, such as lubricants such as magnesium stearate, disintegrants such as calcium cellulose glycolate, and stabilizers such as lactose. , Glutamic acid or aspartic acid. If necessary, tablets or pills may be coated with sugar such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose or the like, or with a film of a gastric or enteric substance. Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like, and commonly used inert diluents, such as purified diluents. Contains water and ethanol. The composition may contain, in addition to the inert diluent, adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, fragrances, and preservatives.

Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Aqueous solutions and suspensions include, for example, distilled water for injection and physiological saline. Examples of water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and polysorbate 80. Such compositions may also include adjuvants such as preserving, wetting, emulsifying, dispersing, and stabilizing agents (eg, lactose) and solubilizing agents (eg, glutamic acid, aspartic acid). May be. These are sterilized by, for example, filtration through a pat- ter filter, blending of a bactericide or irradiation. In addition, these can be used by preparing a sterile solid composition and dissolving in sterile water or a sterile injection solvent before use. (Example)

Next, a production example of the compound of the formula (I) according to the first invention will be described in Examples 1 to 10 described below, and a production example of the compound of the formula (II) according to the second invention will be described later. Examples 11 to 15 will be specifically described. However, the present invention is not limited to only these examples.

Example 1

(a) Synthesis of 2 ', 4'-di-O-acetyl MT 20-dimethyl acetal (Compound 1)

My Power Minosyltylonolide (MT) 20—Dimethyl acetal (2.183 g, 3.38 mmol I) was dissolved in 4 Om I of acetonitrile anhydride, and 0.96 ml of acetic anhydride (10.14 mmol I) was dissolved. , 3 moI eq.) And stirred for 1.5 hours at room temperature for O-acetylation. After completion of the reaction, the solvent was concentrated under reduced pressure from the reaction solution, and the obtained residue was dissolved in 20 OmI of toluene and washed with saturated aqueous sodium hydrogen carbonate. The toluene solution was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and dried to give 2.576 g of the title compound 1 as a yellowish white solid.

(b) Synthesis of 2 ', 4' di-O-acetyl-23-O-t-butyldimethylsilyl-MT 20-dimethylacetal (Compound 2)

Compound 1 obtained in the above (a) was dissolved in 40 ml of anhydrous DMF, and 690.6 mg of imidazole and 765 mg of t-butyldimethylsilyl chloride were added, and 23-O-t-butyldimethyl was added. It was left at room temperature for 5 hours for silylation.

After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel one column chromatography (eluted with toluene monoacetate = 2: 1, silica gel 250 g, 0 = 3.4 cm × 70 cm) to give the title compound 2 as a white solid. 236 g (78%) were obtained.

(c) Synthesis of 2 ', 4'-di-acetyl- 23-O-t-butyldimethylsilyl-13-O-methylthiomethyl-MT 20-dimethyl acetal (Compound 3) Obtained in the previous section (b) Compound 2 (35. Omg) was dissolved in a mixture of acetic anhydride and acetic acid (0.12 ml-0.08 ml), and 0.16 ml of anhydrous dimethylsulfoxide was added to dissolve. When the resulting mixture was left at room temperature for 2 hours, 3-O-methylthiomethylation reaction occurred. After completion of the reaction, the reaction solution was concentrated under reduced pressure. Silica gel obtained —Purification by column chromatography (elution with toluene-acetone = 9: 1, silica gel 6.0 g) gave 26.3 mg (70%) of compound 3. [Shed] _{^{D 25 - 22 ° (c 1}} , CHCI 3).

(d) Synthesis of 23-〇-tert-butyldimethylsilyl-3-O-methylthiomethyl_MT 20-dimethylacetal (compound 4)

Compound 3 obtained in (c) above was dissolved in 1.26 ml of anhydrous methanol and heated at 50 ° C for 7 hours. A deacetylation reaction occurred. After the completion of the reaction, the reaction solution was concentrated under reduced pressure and dried to obtain 192. mg of the title compound 4.

(e) Synthesis of 3-O-methylthiomethyl-MT 20-dimethyl acetal (compound 5)

Compound 4 (192 mg) obtained in (d) above was dissolved in 1.14 ml of tetrahydrofuran (THF), and 0.1 ml of 1 M tetrabutylammonium fluoride THF solution was added. For 1 hour. The completion of the reaction was confirmed by TLC (form of form: methanol / methanol / 28% aqueous ammonia = 10: 1: 0.1). Thereafter, the reaction solution was concentrated under reduced pressure. The obtained syrup was purified by silica gel single column chromatography (eluted with silica gel form-methanol-28% aqueous ammonia = 10: 1: 0.1, silica gel 6.0 g) to give the title compound 5 (47). Omg was obtained.

(f) Synthesis of 3-O-methylthiomethyl-MT [Compound (Ia-1) of the present invention]

47 mg of compound 5 obtained in (e) above was dissolved in 0.24 ml of 1,4-dioxane, 0.24 ml of 3% sulfuric acid was added, and the mixture was stirred and left at room temperature for 1 hour. _{c To the} reaction mixture was added saturated aqueous sodium bicarbonate to adjust the pH to about 9, followed by extraction with chloroform. The chloroform extract was dehydrated with anhydrous sodium sulfate. The obtained organic layer was concentrated under reduced pressure, and the residue was purified by silica gel-one column chromatography (eluted with chloroform-methanol-28% aqueous ammonia = 15: 1: 0.1, silica gel 4.8 g) to give a table. 39.7 mg (96%) of the title compound (Ia-1) was obtained. [H] _D ²⁵ = _45 ° (c1, CHCI ₃ ) _c Calculated (Cssh ^ NOwS); C = 58.55%, H = 8.57%, N = 2.12%, S = 4.87%

Measured value: C = 58.83%, H = 8.53%, N = 2.09%, S = 4.66%

¹ H—NMR spectrum (500 MHz, CDCI ₃ ):

Main signal 5 (ppm): 2.49 (s, NMe2), 4.83 (s, 〇CH2S), 2.13 (s, SMe).

Example 2

(1) Synthesis of 2 ', 4'-di-O-acetyl-23-〇-tert-butyldimethylsilyl-1_3-O-ethylthiomethyl-MT 20-dimethylacetal (Compound 6) Example 1 (b) )), I.e., 2 ', 4' di-〇- ァ -acetyl-23-0-t-butyldimethylsilyl_MT 20-dimethylacetal (451.9 mg) of the formula (V-1), Ethyl methyl sulfoxide (2 ml) was dissolved in a mixture of acetic anhydride and monoacetic acid (1.5 ml—1 ml) and heated at 50 ° C. for 3.5 hours. After confirming the completion of the reaction, the reaction solution was concentrated. The obtained residue was purified by silica gel one-stroke ram chromatography (ethyl ethyl acetate: 2: 1, silica gel 11.2.5 g) to obtain Compound 6 (342.2 mg, 70%). [A] _{^{D 23 - 23 ° (c1,}} CHCI 3).

(2) Synthesis of 23-O-t-butyldimethylsilyl-1-3-ethylethylthiomethyl-MT20-dimethylacetal (compound f)

Compound 6 (207.6 mg) obtained in the above section (1) was dissolved in 4 ml of anhydrous methanol and heated at 50 ° C for 7 hours. After confirming the completion of the de-Asechiru reaction, the reaction solution was concentrated under reduced pressure, and dried to give 1 92. 7m _g of compound 7.

(3) Synthesis of 3-O-ethylthiomethyl-MT 20-dimethyl acetal (compound 8)

Dissolve 192 mg of compound 7 obtained in (2) above in 3.39 ml of THF, add 0.464 ml of 1 M tetrabutylammonium fluoride THF solution, and leave at room temperature for 30 minutes did. The completion of the reaction was confirmed by TLC (cloth form-methanol-28% aqueous ammonia = 10: 1: 0.1). Thereafter, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel single column chromatography (chloroform-methanol-28% aqueous ammonia = 15: 1: 0.1, silica gel 28.5 g) to give 827. 6 mg were obtained. (4) Synthesis of 3-diethylthiomethyl_MT [Compound (Ia_2) of the present invention] 127 mg of Compound 8 obtained in (3) above was dissolved in 0.6 ml of acetonitrile, and further 0.1 M hydrochloric acid was added. 2. 54 ml was added, and the mixture was stirred and left at room temperature for 2 hours. The completion of the deacetation reaction was confirmed by TLC (chloroform / methanol / 28% aqueous ammonia: 10: 1: 0.1). Next, saturated sodium bicarbonate solution was added to the reaction solution to adjust the pH to about 9, followed by extraction with chloroform. The organic layer was dried over anhydrous sodium sulfate, and the organic layer was concentrated under reduced pressure. The resulting residue was subjected to silica gel high-performance column chromatography (chloroform-form-methanol-28% aqueous ammonia = 15: 1: 0.1, silica gel (co-gel C-300) 14 g) to give the title. Compound (Ia-2) (75.5 mg, 63%) was obtained. [H] _D ^{2 1} = —34 ° (c1, CHCI ₃ ). Also, 9.8 mg (8%) of compound 8 and 16.69 mg (15%) of MT were recovered.

Example 3

(1) Synthesis of 2 ', 4'-di-O-acetyl-23-O-t-butyldimethylsilyl-13-O-n-hexylthiomethyl-MT 20-dimethylacetal (Compound 9) 1 Dissolve 37 mg of compound 2 obtained in (b) and n-hexylmethylsulfoxide anhydrous (2 ml) in a mixture of acetic anhydride-acetic acid (1.02 ml — 0.68 ml). Heated at 50 ° C for 4.5 hours. After confirming the end of the reaction by TLC (toluene monoacetate = 2: 1), the reaction solution was concentrated. The residue was purified by silica gel column chromatography (ethyl ethyl toluene acetate: 2: 1, silica gel 74.0 g) to obtain 499.7 mg of the title compound 9 as a yellow syrup.

(2) Synthesis of 23-0-t-butyldimethylsilyl-l-3-0-Γ> -hexylthiomethyl-MM20-dimethylacetal (Compound 10)

To 499 mg of compound 9 obtained in the above section (1), 8 ml of anhydrous methanol was added, and the mixture was heated at 50 ° C for 7 hours. After confirming the completion of the deacetylation reaction by TLC (toluene monoacetate = 1: 1), the reaction solution was concentrated under reduced pressure. The resulting syrupy residue was purified with silica gel Ichiriki column chromatography (toluene monoacetate Echiru = 1: 1; 250 m I one black port formate beam - methanol - ₂ 8% aqueous ammonia = 1 5: 1: 0. 1, Purification on silica gel (75 g) yielded compound 10 (217.8 mg, 60%). [H]. ²⁴ = —28 ° (c1, CHCI ₃ ) _c

(3) Synthesis of 3-O-n-hexylthiomethyl-MT 20-dimethyl acetal (Compound 11) 21.9 mg of the compound 10 obtained in (2) above was dissolved in 3.94 ml of THF, 0.438 ml of a 1 M tetrabutylammonium fluoride THF solution was added, and the mixture was added at room temperature for 1.5 hours. I left it. The end of the desilylation reaction was confirmed by TLC (cloth form-methanol-28% ammonia water = 15: 1: 0.1). Thereafter, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel Ichiriki ram chromatography i (chloroform one methanolate one root _280/0 Anmonia water = 1 5: 1: 0. 1, silica gel 1 7. 6 g) purified by Then, Compound 1 1 1 84.5 mg were obtained.

(4) Synthesis of 3-〇-n-hexylthiomethyl-MT [Compound (Ia-3) of the present invention]

184 mg of the compound 11 obtained in the above section (3) was dissolved in 0.9 ml of acetonitrile, 4 ml of 0.1 M hydrochloric acid was added, and the mixture was stirred and left at room temperature for 2 hours (pH = 1). A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution to adjust the pH to about 9, followed by extraction with chloroform. The organic layer was dried over anhydrous sodium sulfate, and the organic layer was dried under reduced pressure. The obtained residue was subjected to silica gel one column chromatography (cloform form-methanol-28% ammonia water 215: 1: 0.1, 34 g of silica gel (ヮ co-gel C-300) 34 g). The title compound (la_3) (15.8 mg, 64%) was obtained. [H]. ²⁴ = —36 ° (c 1, CHCI ₃ ).

In addition, 8.7 mg of the mixture of compound 11 and compound (Ia-3) and 11.2 mg (8%) of MT were recovered.

Example 4

(1) Synthesis of 2 ', 4'-di-O-acetyl-23-O-t-butyldimethylsilyl-13-O-cyclohexylthiomethyl-MT 20-dimethyl acetal (compound 12)

327.1 mg of compound 2 obtained in Example 1 (b) and cyclohexylmethylsulfoxide (2 ml) were dissolved in a mixed solution of acetic anhydride monoacetic acid (1.02 ml — 0.68 ml). And heated at 50 ° C. for 4 hours. After confirming the completion of the reaction by TLC (toluene-acetone = 5: 2), the reaction solution was concentrated. The residue was purified by silica gel 1 column chromatography (ethyl ethyl toluene acetate = 2: 1, silica gel 132 g) to obtain a solid containing compound 12 (275.9 mg). Further, this was purified by silica gel one column chromatography (toluene monoacetate = 1: 1, silica gel 2 f g) to obtain compound 12 (215.1 mg, 57%). [A] _D ² , = 23 ° (c1, CHCI ₃ ). (2) Synthesis of 23-O-t-butyldimethylsilyl-13-O-cyclohexylthiomethyl-MT20-dimethylacetal (Compound 13)

156.1 mg of the compound 12 obtained in the above section (1) was dissolved in 3 ml of anhydrous methanol and heated at 50 ° C for 7 hours. The reaction solution was concentrated under reduced pressure and dried to obtain 149.3 mg of compound 13.

(3) Synthesis of 3-dicyclohexylthiomethyl-MT 20-dimethyl acetal (compound 14)

Dissolve 149 mg of the compound 13 obtained in the above section (2) in 2.95 ml of THF, add 0.0486 ml of 1 M tetrabutylammonium fluoride in THF, and add 1.0 mol at room temperature. Left for 5 hours. The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel-column chromatography (chloroform-methanol-28% aqueous ammonia = 15: 1: 0.1, silica gel 18 g) to obtain 15.0 mg of compound 14 I got

(4) Synthesis of 3-O-cyclohexylthiomethyl-MT [Compound (Ia-4) of the present invention]

115 mg of compound 14 obtained in the above section (3) was dissolved in 0.58 ml of acetonitrile, and 2.3 ml of 0.1 M hydrochloric acid was added. After stirring, the mixture was allowed to stand at room temperature for 3 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction solution to adjust the pH to about 9, followed by extraction with chloroform (1 ml × 5), and the organic layer was dried over anhydrous sodium sulfate. This organic layer was concentrated under reduced pressure. The residue obtained was purified by silica gel single column chromatography (cloform form-methanol-28% ammonia water = 15: 1: 0.1, 18 g silica gel (ヮ co-gel C-300) 18 g). Compound (Ia-4) (84.3 mg, 73%) was obtained. [H]. ²¹ = —35 ° (C 1, CHCI ₃ ).

Example 5

(1) Synthesis of 2 ', 4'-di-O-acetyl-l23_0-t-butyldimethylsilyl-13-O-phenylthiomethyl-MT 20-dimethylacetal (Compound 15) Example 1 (b) 187.3 _mg of the compound 2 obtained in the above) and phenylmethylsulfoxide anhydride (1 ml) were dissolved in a mixture of acetic anhydride monoacetic acid (0.5 1 ml — 0.34 ml). And heated at 50 ° 0 for 26.5 hours. After confirming the completion of the reaction by TLC (toluene monoacetate = 1: 1), the reaction solution was concentrated. The residue is purified by silica gel lamb chromatography. Purification by GRAPHICS I (toluene monoacetate = 3: 2, silica gel 27.0 g) gave 188. Omg of a mixture containing compound 15. Further, this was purified by silica gel single column chromatography (toluene monoacetate = 5: 2, silica gel 27.O g) to obtain 76.3 mg (38%) of compound 15. [] _D ²³ = _40 ° (c1, CHCI ₃ ).

(2) Synthesis of 23-O-t-butyldimethylsilyl _3-〇_phenylthiomethyl_MT 20-dimethylacetal (compound 16)

241.4 mg of the compound 15 obtained in the preceding section (1) was dissolved in 4.8 mI of anhydrous methanol, and left in an oil bath at 50 ° C. for an hour. The reaction solution was concentrated under reduced pressure and dried to obtain 221.8 mg of compound 16.

(3) Synthesis of 3-O-phenylthiomethyl-MT 20-dimethyl acetal (compound 17)

221 mg of the compound 16 obtained in (2) above was dissolved in 4.4 ml of THF, 0.1 ml of a 1 M tetrabutylammonium fluoride THF solution was added, and the solution was added at room temperature. Left for hours. The reaction mixture was concentrated under reduced pressure and purified by silica gel single column chromatography (cloform form-methanol-28% aqueous ammonia = 15: 1: 0.1, silica gel 40 g) to give compound 16 of 140. 3 mg was obtained.

(4) Synthesis of 3-O-phenylthiomethyl-MT [Compound (Ia-5) of the present invention] 14 Omg of compound 17 obtained in the above (3) was dissolved in 0.7 ml of acetonitrile. Then, 2.8 ml of 0.1 M hydrochloric acid was added, and the mixture was stirred and left at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was dissolved in 7 ml of chloroform and washed with saturated aqueous sodium hydrogen carbonate. The organic layer was dehydrated with anhydrous sodium sulfate and dried under reduced pressure. The residue obtained was subjected to silica gel gel chromatography (chloroform / methanol / 28 <½ammonia water = 15: 1: 0.1, silica gel (ヮPurification with Cogel C-1 300) 21 g) gave 112.3 mg (85%) of compound (Ia-5). [] _D ²¹ = -59 ° (c1, CHCI ₃ ). Example 6

(a) Synthesis of 2'-O-Acetyl_4'-Doximeylminosyltylonolide 20-Dimethylacetanol (Compound 18)

4'-Deoxyximinominosyltylonolide (4'-DMT) 20-Dimethylacetanol (2.043 g, 3.25 mmoI) is dissolved in 4 OmI of acetonitrile anhydride and 0.67 m of acetic anhydride l (10.14mmoI) and add at room temperature. Stirred. After 1.5 hours, completion of the reaction was confirmed by TLC (toluene-acetone = 1: 1), and then the reaction solution was concentrated under reduced pressure. The obtained residue was dissolved in saturated aqueous sodium bicarbonate (8 OmI) and extracted with chloroform. The chloroform extract was dehydrated with sodium sulfate, concentrated under reduced pressure, and dried to obtain 2.546 g of compound 18 as a yellowish white solid.

(b) Synthesis of 2'-diacetyl 23_0-t-butyldimethylsilyl-1 4'-DMT 20-dimethylacetal (compound 19)

Dissolve 2.54 g of compound 18 obtained in the previous section (a) in 40 ml of anhydrous DMF, add 433 mg of imidazole and 7200.3 mg of t-butyldimethylsilyl chloride, and at room temperature for 5 hours. I left it. The reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel one-column chromatography (toluene-acetone = 5: 2, silica gel 200 g) to obtain 2.245 g (87%) of compound 19 as a white solid.

(c) Synthesis of 2'_〇-acetylacetyl 23_0-t-butyldimethylsilyl-13-O-methylthiomethyl-14'-DMT20-dimethylacetal (Compound 20) Compound 1 obtained in the previous section (b) 9 502.6 mg and 2 m of anhydrous dimethyl sulfoxide

I was dissolved in a mixed solution of acetic anhydride and monoacetic acid (1.5 mI—1 mI) and left at room temperature for 72 hours. The completion of the reaction was confirmed by TLC (toluene-acetone = 9: 1). The reaction solution was concentrated under reduced pressure, and the obtained syrup was purified by silica gel single column chromatography_ (toluene-acetone = 5: 2, silica gel 45 g) to obtain 427 mg (8%) of compound 20. [H] _D ²¹ = -2 ° (c1, CHCI ₃ ).

(d) Synthesis of 23—O—t—butylmethylsilyl-1-O—methylthiomethyl-14′—DMT20—dimethyl acetal (Compound 21)

15 Omg of compound 19 obtained in the above section (c) was dissolved in 3.0 ml of anhydrous methanol and heated at 50 ° C for 5 hours. The completion of the deacetylation reaction was confirmed by TLC (toluene-acetone = 1: 1). Then, the reaction solution was concentrated under reduced pressure and dried to obtain 142.3 mg of compound 21.

(e) Synthesis of 3_-methylthiomethyl-14'-DMT20_dimethylacetal (Compound 22)

142 mg of the compound 21 obtained in the above (d) was dissolved in 2.85 ml of THF, 0.25 ml of a 1 M tetrabutylammonium fluoride THF solution was added, and the solution was added at room temperature. . Left for 5 hours. TLC (Black mouth form-methanol'-28% ammonia The completion of the desilylation reaction was confirmed with water = 10: 1: 0.1). The syrup obtained by concentrating the reaction solution under reduced pressure was purified by silica gel single column chromatography (chloroform / methanol / 28% aqueous ammonia 10: 1: 0.1, silica gel 14 g) to give compound 22. 97.2 _mg was obtained.

(f) Synthesis of 3-P-methylthiomethyl-14'-DMT [Compound (Ib_1) of the present invention]

97.2 mg of the compound 22 obtained in (e) above was dissolved in 0.49 ml of 1,4-dioxane, and 3 V V <0.49 ml of sulfuric acid was added. After stirring, the mixture was stirred at room temperature. For 1 hour. To the reaction solution was added a saturated aqueous sodium hydrogen carbonate solution to adjust the pH to about 9, followed by extraction with chloroform (1 m IX 5), and the organic layer was dried over anhydrous sodium sulfate. The obtained organic layer was dried under reduced pressure, and the obtained residue was subjected to silica gel-one column chromatography (chloroform-form-methanoyl-28% aqueous ammonia = 15: 1: 0.1, silica gel 9.8 g). Purification gave 66.3 mg (73%) of compound (Ib-1). [Α] _D ²⁵ = - 47. (C 1, CHCI _3). Example 7—10

(1) The 2′-O-acetyl-23-O-t-butyldimethylsilyl-1 4′-DMT 20-dimethylacetal (compound 19) obtained in Example 6 (b) was converted to acetic anhydride-acetic acid

(Approximately 3: 2 by volume) at 50 ° C in the same manner as in Example 6 (c) in the same manner as in Example 6 (c), except for anhydrous ethyl methylsulfoxide, n-hexylmethylsulfoxide, cyclohexylmethylsulfoxide or phenyl. Reacted with methylsulfoxide.

At this time, 502.6 mg of Compound 19 was reacted with 3 ml of anhydrous ethylmethylsulfoxide, 374.2 mg of Compound 19 was reacted with 2 ml of anhydrous n-hexylmethylsulfoxide. 190.4 mg of compound 19 was reacted with 1 ml of anhydrous cyclohexylmethylsulfoxide. Then, 370.4 mg of compound 19 was reacted with 2 ml of anhydrous phenylmethylsulfoxide. The amount of the acetic anhydride-acetic acid mixture used was in the range of about 3 mI to about 1 mI.

After confirming the completion of the reaction by TLC, the reaction solution was post-treated in the same manner as in Example 6 (c). This gives the compounds summarized in Table 3 below as 2′-〇-acetyl-23_〇_t-butyldimethylsilyl-13- 一 -substituted thiomethyl-4 ′ of the formula (VI;). -DMT 20-dimethyl acetal was obtained. (V,)

Table 3

(2) Each of the compounds 23 to 26 obtained in the above (1) was heated at 50 ° C. in anhydrous methanol in the same manner as in Example 6 (d) to perform a deacetylation reaction. . The reaction solution was post-treated in the same manner as in Example 6 (d). Thus, as a compound summarized in Table 4 below, 23-O-t-butyldimethylsilyl-13-O-substituted thimethyl-4'-DMT20-dimethyl of formula (VII:;) Acetal was obtained.

(V,)

Table 4

(3) Each of Compounds 27 to 30 obtained in (2) above was reacted with 1 M tributylammonium fluoride in THF at room temperature in the same manner as in Example 6 (e). Thus, a desilylation reaction was performed. After confirming the completion of the reaction by TLC, Post-treatment was carried out in the same manner as in Example 6 (e).

As a result, 3〇-substituted thiomethyl-14′-DMT 20 _dimethylacetal of the formula (IX ′) was obtained as a compound summarized in Table 5 below.

Table 5

Two

Three

(4) Each of the compounds 31 to 34 obtained in the above item (3) is dissolved in about 0.35 ml to 0.85 ml of acetonitrile, and further, about 1.3 m I to 3.5 m of 1 M hydrochloric acid. After adding l and stirring, the mixture was allowed to stand at room temperature for 2 to 4 hours. A deacetalization reaction was performed by hydrolysis. After confirming the completion of the reaction by TLC, the reaction solution was post-treated in the same manner as in Example 6 (f). This gave 3 —O—substituted thiomethyl-14′—DMT of formula (I b ′) as compounds summarized in Table 6 below.

(lb ')

Table 6

In addition to the above examples, other compounds of the present invention are shown below in Tables 7 and 8.

These compounds can be synthesized using the synthetic routes and methods described in the above Preparations and Examples, and modifications thereof, which are generally known to those skilled in the art, and require special experimentation. It does not mean that.

Front

Table 8

H

Claims

, ^: Claims The following general formula (I)

[In the formula, X represents a hydroxyl group or a hydrogen atom, R represents a group Ch ^ SR ¹ (where R ¹ represents an alkyl group, a cycloalkyl group or an aryl group), and Me represents a methyl group.] Or a salt thereof.

2. The myominosyltylonolide derivative or a salt thereof according to claim 1, wherein X is a hydroxyl group.

3. A pharmaceutical composition comprising, as an active ingredient, the myo-minosyltylonolide derivative according to claim 1 or a biologically acceptable salt thereof.