KR950010086B1 - New 2'-substituted-4-deoxythiazolo(5,4-c)rifamycin sv derivatives - Google Patents

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Description

New 2′-Substituted-4-deoxy-thiazolo [5,4-c] rifamycin SV Derivatives

The present invention relates to novel novel 2′-substituted-4-deoxy-thiazolo [5,4-c] rifamycin SV derivatives represented by the following general formula (I) and pharmaceutically acceptable acid addition salts thereof.

[식 중, R⁴는 수소, (C₁-C₄)알킬, 히드록시(C₂-C₄)알킬, (C₂-C₄)알카노일, (C₁-C₄)알콕시 카르보닐, 페닐(이 페닐기는 할로게노, 히드록시, (C₁-C₄)알킬 및 (C₁-C₄)알콕시기 중에서 선택되는 1 내지 3개의 치환체로 임의 치환될 수 있음), 페닐(C₁-C₄)알킬(여기에서, 페닐 고리는 할로게노, 히드록시, (C₁-C₄)알킬 및 (C₁-C₄)알콕시기 중에서 선택되는 1 내지 3개의 치환체로 임의 치환 될 수 있음) 및 페닐(C₁-C₄)알콕시카르보닐기(여기에서, 페닐 고리는 할로게노, 히드록시, (C₁-C₄)알킬 및 (C₁-C₄)알콕시기 중에서 선택되는 1 내지 3개의 치환체로 치환될 수 있음)임] 중에서 선택되는 헤테로기를 추가로 함유할 수도 있다.Wherein R ¹ is hydrogen or an acetyl group, R ² and R ³ are each independently a (C ₁ -C ₄ ) alkyl group, or R ² and R ³ are a 4-7 membered saturated hetero with an adjacent nitrogen atom Cyclic rings, where the rings are coral, sulfur and

[Wherein R ⁴ is hydrogen, (C ₁ -C ₄ ) alkyl, hydroxy (C ₂ -C ₄ ) alkyl, (C ₂ -C ₄ ) alkanoyl, (C ₁ -C ₄ ) alkoxycarbonyl, Phenyl (this phenyl group may be optionally substituted with 1 to 3 substituents selected from halogeno, hydroxy, (C ₁ -C ₄ ) alkyl and (C ₁ -C ₄ ) alkoxy groups), phenyl (C _1- C ₄ ) alkyl, wherein the phenyl ring may be optionally substituted with one to three substituents selected from halogeno, hydroxy, (C ₁ -C ₄ ) alkyl and (C ₁ -C ₄ ) alkoxy groups) And a phenyl (C ₁ -C ₄ ) alkoxycarbonyl group, wherein the phenyl ring is 1 to 3 substituents selected from halogeno, hydroxy, (C ₁ -C ₄ ) alkyl and (C ₁ -C ₄ ) alkoxy groups It may be substituted with)) may further contain a hetero group selected from.

4-deoxy-thiazolo [5,4-c] rifamycin SV (also referred to as "rifamycin P") has been obtained by fermenting Nocardia strains or by chemical methods. Fermentation methods are described in British Patent No. 1,470,426 and chemical methods are described in US Pat. Nos. 4,144,234 and 4,129,562.

2'-alkyl-4-deoxy-thiazolo [5,4-c] rifamycin SV derivatives are described in US Pat. No. 4,169,834 and described as 2'-hydrazonomethyl-4-deoxythiazolo [5, 4-c] rifamycin SV is described in European Patent Publication No. 5140, and 4-deoxythiazolo [5,4-c] rifamycin SV with one amino substituent or hydrazino substituent at the 2 'position It is described in Federal Republic of Laid-Open Patent Application 27 41 066, which is prepared by reacting 3-bromo rifamycin S with a suitable thiourea derivative.

중에서 선택되는 헤테로기를 추가로 함유할 수 있는 4,5,6 또는 7원 포화 헤테로 시클릭 고리를 의미한다. 이 4-7원 헤테로시클릭 고리로서 적합한 예는 아제티디닐, 피롤리디닐, 피페리디닐, 피페라지닐, 옥사졸리니디닐, 피라졸리디닐, 티아졸리디닐, 모르폴리닐, 이미다졸리디닐 등이다.The term “(C ₁ -C ₄ ) alkyl” in this specification and in the claims, whether alone or in combination with other groups, refers to methyl, ethyl, propyl, 1-methylethyl, 2-methyl-1-propyl, 2- It means an alkyl group having 1 to 4 carbon atoms such as methyl-2-propyl and butyl. The term “(C ₁ -C ₄ ) alkoxy”, alone or in combination with other groups, refers to methoxy, ethoxy, propoxy, 1-methylethoxy, 2-methoxypropoxy, 2-methyl-2-propoxy and It means an alkoxy group of 1 to 4 carbon atoms, such as butoxy. The term “halogen” refers to a halogen atom selected from chloro, bromo, iodine and fluoro. “4-7 membered saturated heterocyclic ring... The term oxygen, sulfur and

It means a 4,5,6 or 7 membered saturated heterocyclic ring which may further contain a hetero group selected from. Examples suitable as this 4-7 membered heterocyclic ring are azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxazolininidinyl, pyrazolidinyl, thiazolidinyl, morpholinyl, imidazolidinyl And so on.

Suitable compounds among the compounds according to the invention are those of the general formula (I) wherein R ¹ is an acetyl group. Another suitable compound is R ² and R ³ are independently (C ₁ -C ₄ ) alkyl groups, or R ² and R ³ together with the adjacent nitrogen atom are azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl , A 4-7 membered heterocyclic ring selected from oxazolininidinyl, pyrazolidinyl, thiazolidinyl, morpholinyl, imidazolidinyl group, and further, when a ring nitrogen atom is present, the nitrogen atom is substituted with R ⁴ ( This is a compound of formula (I) containing as defined above.

Further suitable compounds among the compounds according to the invention are those wherein R ¹ is an acetyl group, R ² and R ³ are independently a methyl or ethyl group, or R ² and R ³ may be optionally substituted with adjacent nitrogen atoms (here In which R ⁴ is as defined above) is a compound of formula (I) representing a pyrrolidinyl, morpholinyl, piperidinyl or piperazinyl group. Specific examples of suitable compounds among the compounds according to the invention include 2 '-(4''-morpholinyl) rifamycin P, 2'-(1 ''-piperidinyl) rifamycin P, 2 '-(4''-Methyl-1'-piperazinyl) rifamycin P and 2'-(NN-diethylamino) rifamycin P.

Compounds according to the invention are 4-deoxy-thiazolo [5,4-c] rifamycin SV (ie rifamycin P) or 25-desacetyl-4-deoxy-thiazolo [5,4-c] Rifamycin SV, ie general formula (II)

Wherein R ¹ is hydrogen or an acetyl group, and is reacted with an amine of the general formula HNR ² R ³ in the presence of an inert organic solvent, provided that R ⁴ when the HNR ² R ³ amine is piperazine Is a group other than hydrogen. In order to obtain a piperazine derivative which is R ⁴ , R ⁴ is subjected to hydrocracking under a condition of preventing hydrogenation of the double bond based on the corresponding phenyl (C ₁ -C ₄ ) alkoxycarbonyl derivative. The temperature of this reaction is generally 5 ° C. to 40 ° C., suitably 15 ° C. to 30 ° C., particularly suitably room temperature. As an inert organic solvent, it is not necessary to completely dissolve during the reaction so that the starting material or the final product is in a homogeneous phase, but adverse effects on the reaction route, which can at least partially dissolve the starting material of Formula 2 and the amine HNR ² R ³ . Any one of the organic solvents that does not affect may be used. Examples of representative inert organic solvents are saturated “cyclic” ethers such as tetrahydrofuran or dioxane, glycol ethers such as dimethoxyethane and (C ₁ -C ₄ ) alkyl such as ethyl acetate, propyl acetate, ethylpropionate and the like. Alkanoate and mixtures thereof.

R ⁴ is and the corresponding compound gateuna other than hydrogen as defined above as a starting material, R ⁴ a hydrocracking reaction steps required to obtain the hydrogen of piperazine derivative is not substantially gyejae induce substantial hydrogenation of the double bond It must be done under conditions that do not. This hydrocracking reaction is suitably carried out in a polar organic like lower alkanol in the presence of a hydrogenation catalyst such as charcoal based palladium. Particularly suitable conditions are those carried out in the presence of anhydrous ethanol as solvent and charcoal based 10% palladium as catalyst. The reaction pathway is monitored by HPLC or TLC techniques in a manner known in the art to enable the skilled person to control certain reaction conditions and determine if the reaction has ended.

To purify the compound according to the present invention, conventional purification such as extraction with a solvent, addition of a non-solvent, or precipitation and chromatographic methods for converting the pH of the medium are suitable. Chromatographic treatments include preparative TLC, column chromatography and HPLC.

The physicochemical data and purification methods of some representative compounds of the compounds according to the invention are set forth in Tables 1 and 2 below. In particular, the following compounds are compounds of the general formula (I) wherein R ¹ is an acetyl group, except compound 3b in which R ¹ is hydrogen.

TABLE 1

[Comment on Table 1]

1) When two quantities were described, the reagents were added in two portions at the indicated time.

2) a) acid treatment as described for compound 1a, b) acid / base treatment as described for compound 3a, c) preparative TLC, eluent mixture CH ₂ Cl ₂ MeOH 95: 5, compound Collected, dissolved in a mixture of EtOAc and acid water, the organic layer was separated, concentrated to a small volume and the product precipitated using petroleum ether, d) CH ₂ Cl ₂ MeOH 95: 5 Preliminary TLC with eluent mixture followed by treatment as described in compound 4, e) crystallized from EtOAc, f) precipitated using petroleum ether from EtOAc, g) petroleum ether from CH ₂ Cl ₂ Precipitated using h) column chromatography using Silica Gel 60 [0.06-0.2mm, Merck Co.] using an eluent mixture of CH ₂ Cl ₂ / MeOH 95: 5.

TABLE 2

[Comment on Table 2]

Solvent elution 10.0 cm on a Silica Gel 60 F ₁₅₄ plate (Merck).

1) The analysis results for C, H, N, S (and Cl) were within ± 0.4% of theory.

2) Eluent Compound CHCl ₃ / MeOH 9/1. Under these conditions, the R _f value of rifamycin P was 0.42.

3) Measured in glass capillary.

4) Lipamycin P showed lambda max values of 225 nm, 260 nm, 305 nm, 352 nm and 412 nm in methanol.

n.d. = Not measured.

The compound according to the present invention has an antimicrobial action against Gram-positive bacteria and some Gram-negative bacteria and is also effective against Mycobacteria strains, in particular Mycobacterium tuberculosis.

The minimum inhibitory concentration (MIC) of a representative compound among the compounds according to the invention was determined using a 2-fold dilution method in the microtiter mode. As medium, Todd-Hewitt broth (product of Difco) for Streptococcus and Iso-Sensitest broth for Straphylococcus and Gram-negative bacteria ) [Oxoid product] was used. The final inoculum was about 10 ⁴ cf / μml. MIC was read as the minimum concentration showing no visible growth after 18-24 hours incubation at 37 ° C. For Mycobacterium tuberculosis H ₃₅ Rv, the inoculum consisted of 0.5-1% of 7-9 days culture in Dubos medium. This test was conducted in Kirschner: Medium supplemented with 10% horse serum. At this time, it was observed after incubation for 7 days at 37 ℃.

The above results are summarized in Table 3 below.

TABLE 3

Antimicrobial activity in vitro (MIC, mg / ℓ)

a) supplemented with 3% bovine serum b) strain resistant to rifampicin c) n.t = not tested

The antimicrobial action of the compounds according to the invention has also been confirmed in vivo. The results of experimental sepsis in piglets are listed in Table 4 below.

Animals (group of five birds) were infected by intraperitoneal injection of a 16 hour juicy culture of Staphylococcus aureus tour L 165. Untreated animals were inoculated to die within 48 hours of sepsis. Animals were treated once daily for 3 days starting immediately after infection. After 10 days, the ED ₅₀ value (mg / kg / day) is based on the percentage of animals surviving at each dose, SPEARMAN-KARBER [Pinney, D. second. (Finney, DJ), Statistical method in biological assay, page 524, Seeds, London. Calculated by C. Griffin Ltd., 1952.

TABLE 4

Hereinafter, the present invention will be described in more detail with reference to Examples.

Example 1

a) 2 '-(1' '-pyrrolidinyl) rifamycin P

10 ml of pyrrolidine was added to a solution of 1.5 g (1.35 mmol) of rifamycin p dissolved in 150 ml of THF at room temperature. The reaction mixture was stirred for 5 hours, then poured into ice water, acidified by the addition of dilute hydrochloric acid, and extracted with ethyl acetate (EtoAc) (100 ml × 2 times). The combined extracts were washed with water until neutral, dried over anhydrous sodium sulfate, and then concentrated to a small volume under vacuum at 45 ° C. Petroleum ether was added thereto to obtain a precipitate, which was collected and purified by preparative TLC as follows. That is, the solid was dissolved in ml of MeOH water, and this solution was added to Silica Gel plate 60PF ₂₅₄ (manufactured by Merck) and developed using a CH ₂ Cl ₂ / MeOH 95: 5 mixture. The area corresponding to the title product was scraped off and eluted with methanol (MeOH). The extract was evaporated to dryness, the residue dissolved in ethyl acetate, washed with dilute hydrochloric acid, dried over sodium sulfate and then concentrated to a small volume. Petroleum ether was added here to produce a precipitate which was filtered off and dried under vacuum. Yields and physicochemical data are shown in Tables 1 and 2 above.

The following compounds were obtained using appropriate amine reactants by the treatment of Example (1a) above, and the reaction conditions, purification methods, yields and physicochemical data of these compounds are listed in Tables 1 and 2 above.

b) 2 '(4' '-morpholinyl) rifamycin P

HNR ² R ³ = morpholine

c) 2 '(4' '-phenyl-1' '-piperazinyl) rifamycin P

HNR ² R ³ = 1-phenylpiperazine

d) 2 '(4' '-phenylmethyl-1' '-piperazinyl) rifamycin P

HNR ² R ³ = 1-benzylpiperazine

e) 2 '(4' '-ethoxycarbonyl-1' '-piperazinyl) rifamycin P

HNR ² R ³ = 1-ethoxycarbonylpiperazine

f) 2 '(4' '-phenylmethoxycarbonyl-1' '-piperazinyl) rifamycin P

HNR ² R ³ = 1-phenylmethoxycarbonylpiperazine

Example 2

2 '-(1' '-piperidinyl) rifamycin P

1.0 g (1.35 mmol) of rifamycin P was dissolved in 15 ml of piperidine, and the resulting solution was stirred at room temperature for 30 hours. The reaction mixture was poured into ice water and treated by the method described in Example (1a) above. Purification, yield and physicochemical data are set forth in Tables 1 and 2 above.

Example 3

a) 2 '-(4' '-methyl-1' '-piperazinyl) rifamycin P

10 ml of 1-methylpiperazine was added to a solution of 2.0 g (2.7 mmol) of rifamycin P dissolved in 300 ml of EtOAc, and the reaction mixture was stirred at room temperature for 90 minutes, followed by 10 ml of 1-methylpiperazine. More was added. The reaction mixture was continuously stirred for 3.5 hours and then poured into ice water, where acidified to maintain pH 4-5 by addition of dilute hydrochloric acid.

The organic layer was discarded and the aqueous phase was brought to about pH 7 by addition of NaHCO ₃ and extracted with EtOAc. The organic extract was dried using sodium sulfate and concentrated to a small volume. This was cooled to obtain the title compound as a precipitated crystalline powder. Purification, yield and physicochemical data are set forth in Tables 1 and 2 above.

The following compounds were obtained using a suitable amine reactant by the treatment of Example (3a) above, the purification, yield and physicochemical data thereof are listed in Tables 1 and 2 above.

b) 25-desacetyl-2 '-(4' '-methyl-1' '-piperazinyl) rifamycin P

(25-desacetyl rifamycin P as starting material)

HNR ² R ³ = 1-methylpiperazine

c) 2 '-[4' '(2' ''-hydroxyethyl) -1 ''-piperazinyl] rifamycin P

HNR ² R ³ = 1- (2-hydroxyethyl) piperazine

d) 2 '-[4' '(4' ''-chlorophenyl) -1-piperazinyl] rifamycin P

HNR ² R ³ = 1- (4-chlorophenyl) piperazine

e) 2 '-[4' '(4' ''-methoxyphenyl) -1-piperazinyl] rifamycin P

HNR ² R ³ = 1- (4-methoxyphenyl) piperazine

Example 4

2 ''-(1 ''-piperazinyl) rifamycin P

A solution of 0.22 g (0.23 mmol) of 2 '-[4''-phenylmethoxycarbonyl-1'-piperazinyl] rifamycin P dissolved in 10 ml of anhydrous ethanol was charcoal based while stirring at room temperature and atmospheric pressure under a stream of hydrogen. It was left for 1 hour in the presence of 50 mg of 10% palladium. The reaction was monitored by TLC (Rf = 0.4 relative to starting compound). At the end of the reaction, the reaction mixture was filtered and then the solvent was evaporated. The residue was dissolved by addition of MeOH and purified by preparative TLC (eluent mixture of CHCl ₃ / MeOH 9: 1) using a Silica Gel 60 PF ₂₅₄ plate. The region containing the title product was eluted using MeOH. The methanol extract was evaporated to dryness and the residue was dissolved by addition of ethyl acetate, washed with aqueous NaHCO ₃ solution at pH 8.5, dried using sodium sulfate and then concentrated to a small volume. After adding petroleum ether here, the title compound was precipitated, which was filtered off and dried under vacuum.

Example 5

2 '-(N, N-diethylamino) rifamycin P

5 ml of N, N-diethylamine was added to a solution of 0.5 g (0.67 mmol) of rifamycin P dissolved in 25 ml of EtOAc, and the reaction mixture was left at room temperature for 5 days, then poured into ice water and diluted hydrochloric acid was added. Made acidic by addition. The organic layer was separated, washed with water until neutral, dried with sodium sulfate and evaporated to dryness. The residue was dissolved by adding ₂ ml of CH ₂ Cl ₂ , and then Silical Gel 60 (0.04-0.06 mm, available from Merck) was added to a 35 g column, and developed by flash chromatography using the following eluent.

a) 250 ml CH ₂ Cl ₂ .

b) 500 ml of CH ₂ Cl ₂ containing 0.5% (v / v) methanol.

c) 500 ml of CH ₂ Cl ₂ containing 1% (v / v) methanol.

d) 1 L of CH ₂ Cl ₂ containing 1.8% (v / v) methanol.

The title product was eluted from this final mixture, which was collected into 12 fractions of 100 ml each. Fractions containing the title product were combined and evaporated to dryness to give 33 mg of pure compound.

[Production of Starting Material]

Rifamycin P (4-deoxy-thiazolo [5,4-c] rifamycin SV)

Rifamycin P was prepared by the methods described in British Patent 1,470,426, United States Patent 4,144,234 or United States Patent 4,129,562.

2'-methoxycarbonyl rifamycin P

2'-methoxycarbonyl rifamycin P was prepared by the method described in Tetrahedron, Cricchio et al., No. 36, pp. 1415-1421 (1981) and U.S. Pat.

25-desacetyl rifamycin P

A solution of 4.5 g (5.5 mmol) of 2'-methoxycarbonyl rifamycin P dissolved in 200 ml of acetone and 100 ml of 50% NaOH aqueous solution was left to stand at room temperature with stirring. The reaction was monitored by TLC (CHCL ₃ / MeOH, 95: 5) until the starting compound disappeared (about 30 minutes). The reaction mixture was poured into iced water and extracted with EtOAc. The extract was dried using sodium sulfate and then evaporated to dryness in vacuo for 30 minutes. The residue was dissolved in 300 ml of the same amount of acetone / NaOH aqueous solution mixture and the solution was evaporated again. The residue was dissolved by addition of CHCl ₃ and purified by Silica Gel (220 g) column chromatography using CHCl ₃ containing increased amount of MeOH (1.5-2% by volume) as eluent. Fractions containing the desired product were combined and the solvent was evaporated to dryness. It was crystallized using EtOAc to give 3.5 g (90%) of the title compound.

Melting Point: 172 ℃ -174 ℃

UV (MeOH) λ max nm (log): 224 (4.26), 260 (4.21), 298 (4.17), 405 (4.14).

Analysis of C ₃₆ H ₄₄ N ₂ O ₁₀ S

Calc .: C 62.05; H 6.36; N 4.02; S 4.60.

Found: C 61.76; H 6.40; N 3.95; S 4.47.

Claims (5)

2'-substituted-4-deoxy-thiazolo [5,4-c] [5,4-c] rifamycin SV derivative represented by the following general formula (I), and its pharmaceutically acceptable acid addition salt thereof. [Formula I]

Wherein R ¹ is hydrogen or an acetyl group, R ² and R ³ are each independently a (C ₁ -C ₄ ) alkyl group, or R ² and R ³ together with an adjacent nitrogen atom, azetidinyl, pi Lolidinyl, piperidinyl, unsubstituted or 4N ''-substituted piperazinyl, wherein the substituents are hydrogen, (C ₁ -C ₄ ) alkyl, hydroxy (C ₂ -C ₄ ) alkyl, (C _1- C ₄ ) alkoxycarbonyl, phenyl, halo substituted phenyl, (C ₁ -C ₄ ) alkoxy substituted phenyl, phenyl (C ₁ -C ₄ ) alkyl and phenyl (C ₁ -C ₄ ) alkoxycarbonyl ], Oxazolinidinyl, unsubstituted or 3N ''-substituted pyrazolidinyl, wherein the substituents are as defined above for the substituents of piperazinyl, thiazolidinyl, morpholinyl, and unsubstituted or 3N ' Represents a saturated heterocyclic ring selected from '-substituted imidazolidinyl, wherein the substituent is as defined for the substituent of piperazinyl above . The compound of claim 1, wherein R ¹ is an acetyl group. The compound of claim 1, wherein R ¹ is an acetyl group, R ² and R ³ are each independently a methyl or ethyl group, or R ² and R ³ together with an adjacent nitrogen atom, pyrrolidinyl, morpholinyl, piperidinyl Or an unsubstituted or 4N ''-substituted piperazinyl, wherein the substituents are as defined in claim 1. 2. The compound of claim 1 wherein 2 ′ (4 ″ -morpholinyl) rifamycin P, 2 ′-(1 ″ -piperidinyl) rifamycin P, 2 ′-(4 ''-methyl-1 '' -Piperazinyl) rifamycin P, and 2 '-(N, N-diethylamino) rifamycin P. A pharmaceutical composition having antimicrobial activity comprising the compound of claim 1 as an active ingredient and a pharmaceutically acceptable carrier.