KR100559494B1 - Novel 1?-methylcarbapenem derivatives, process of preparation thereof and pharmaceutical composition comprising the same for antibiotics - Google Patents

Abstract

The present invention relates to a novel 1-betamethylcarbapenem derivative, a preparation method thereof, and a pharmaceutical composition for antibiotics containing the same as an active ingredient, wherein the 1-betamethylcarbapenem derivative is No. 2 of 1-betamethylcarbapenem nucleus. And a 5- (3'-alkyloxyimino) pyrrolidinylamidopyrrolidine-3-thio group as the main functional group at the position.

Since the 1-betamethylcarbapenem derivative of the present invention shows excellent antimicrobial activity against Gram-negative bacteria including not only Gram-positive bacteria but also ESBL (extended-spectrum β-lactamase) and MDR (multi-drug resistant) producing strains, It can be usefully used.

Description

Novel 1β-methylcarbapenem derivatives, process of preparations and pharmaceutical composition comprising the same for antibiotics

The present invention relates to a novel 1-betamethylcarbapenem derivative, a preparation method thereof and a pharmaceutical composition for antibiotics containing the same as an active ingredient.

Carbapenem antibiotics exhibit a broader antimicrobial activity than conventional antibiotics cephalosporin or penicillin, as well as excellent effects against resistant bacteria. However, these carbapenem antibiotics are easily decomposed and inactivated by the enzyme dehydropeptidase-I (abbreviated as "DHP-I"), which is present in the kidney, and thus lose antimicrobial activity. ( J. Antibiot ., 1991 , 1172-1177, Antimicrobial Agent and Chemotherapy, 1992 , 1577-1579).

Meanwhile, 1-betamethylcarbapenem compounds exhibit broad antimicrobial spectrum against both Gram-positive and Gram-negative bacteria, as well as stability against DHP-I, resulting in meropenem having a 1-betamethyl group in the carbapenem nucleus. And commercially relevant derivatives, BO-2727, S-4661, ZD-4433, ER-35786, FR-21818 and IH201, are in clinical or animal testing ( J. Antibiot ., 1990 , 43 , 519, Yamaji). , E. et al . Abstracts of Papers, H141; 35th Interscience Conference on Antimicrobial Agents and Chemotheraphy, San Francisco, CA, 17-20 Sep. 1995, Arakawa, S. et al . Abstracts of Papers, F218; 37th Interscience Conference on Antimicrobial Agents and Chemotheraphy, Toronto, Ontario, 28 Sep.-1 Oct. 1997, Pelak, BA et al . Abstracts of Papers, F119; 36th Interscience Conference on Antimicrobial Agents and Chemotheraphy, New Orleans, LA, 15-18 Sep. 1996 , Sato, N. et al . Abstracts of Papers, F151; 35th Intersci ence Conference on Antimicrobial Agents and Chemotheraphy, San Francisco, CA, 17-20 Sep. 1995, Tawara, S. et al . Abstracts of Papers, F145; 35th Interscience Conference on Antimicrobial Agents and Chemotheraphy, San Francisco, CA, 17-20 Sep. 1995, Shin, KJ et al . Bioorg. Med. Chem. Lett. 1998 , 8 , 1607.).

Thus, the present inventors synthesized a new 1-betamethylcarbapenem derivative in order to develop antibiotics having superior antimicrobial activity than conventional antibiotics, and showed high stability against DHP-I in these compounds, but also produced Gram-positive bacteria and ESBL and MDR. It was confirmed that there is an excellent antimicrobial activity against all Gram-negative bacteria including the strain and completed the present invention.

The present invention seeks to provide novel 1-betamethylcarbapenem derivatives.

In addition, the present invention is to provide a method for preparing the 1-betamethylcarbapenem derivative.

The present invention also provides a pharmaceutical composition for antibiotics containing the 1-betamethylcarbapenem derivative as an active ingredient.

In the present invention, 5- (3'-alkyloxyimino) pyrrolidinylamidopyrrolidine-3-thio group is substituted as the main functional group at position 2 of the 1-betamethylcarbapenem nucleus represented by the following general formula (1). Provided are 1-betamethylcarbapenem derivatives.

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.)

Preferred compounds among the 1-betamethylcarbapenem derivatives of the present invention are specifically as follows.

1) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6- [(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid

2) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6- [ (1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

3) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio ] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

4) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio ] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

5) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-carboxypyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

6) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-ethoxycarbonylpyrrolidin-1-ylcarbonyl) pyrrolidine-3- Ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid

The 1-betamethylcarbapenem derivative of the present invention represented by Chemical Formula 1 may be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful as a salt. Do. Inorganic acids and organic acids can be used as the free acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. can be used as the inorganic acid, citric acid, acetic acid, lactic acid, maleic acid, umarin acid, gluconic acid, methanesulfonic acid , Glyconic acid, succinic acid, 4-toluenesulfonic acid, trifluoroacetic acid, galluxuronic acid, embonic acid, glutamic acid, aspartic acid and the like can be used.

In addition, the present invention provides a method for preparing a 1-betamethylcarbapenem derivative of Formula 1, which is represented by Scheme 1 below.

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.)

Method for preparing 1-betamethylcarbapenem derivative of the present invention

1) preparing a carbapenem derivative (XII) by reacting a carbapenem enol phosphate compound (II) with a thiol derivative (III) in the presence of a base (first step), and

2) a deprotection reaction of the carbapenem derivative (XII) obtained in the first step to prepare 1-betamethylcarbapenem derivative (I) (second step).

In the first step, carbapenem enol phosphate compound (II) used as starting material was prepared by a known method (Catchpole, CR et al . Antimicrob. Agents Chemother. 1992 , 36 , 1928).

In more detail, in the first step, the base may use a tertiary organic amine having a low basicity, preferably trimethylamine, triethylamine, diisopropylethylamine, 2,6- Lutidine, picoline, N , N'-dimethylaniline, pyridine, 4-dimethylaminopyridine can be selected and used.

The reaction is carried out by reacting the carbapenem enol phosphate compound (II) and thiol derivative (III) in acetonitrile solution using diisopropylethylamine as a base at 0 ° C. for 3 hours to obtain a carbapenem derivative (XII).

In the second step, the carbapenem derivative (XII) obtained in the first step is used for 2 hours at 0 ° C. using tributyltinhydride and tetrakis (triphenylphosphine) palladium (0) catalyst in methylene chloride solution. Reaction gives the desired 1-betamethylcarbapenem derivative (I) deprotected.

The present invention also provides a thiol derivative (III) represented by the following formula (2) as an intermediate.

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.)

In addition, the present invention provides a method for preparing a thiol derivative (III) of Formula 2, which is represented by the following Scheme 2.

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.)

Method for preparing a thiol derivative (III) represented by the formula (2) used to prepare the 1-betamethylcarbapenem derivative of the present invention

1) A pyrrolidinone compound (X) and a hydroxylamine or methoxyamine compound (XI) protected with a butoxycarbonyl group were prepared by using sodium hydrogencarbonate as a base in a mixed solution of ethanol and tetrahydrofuran (2: 1). Reaction at ˜40 ° C. for 1 hour to produce pyrrolidine compounds, which were first reacted with trifluoroacetic acid and triethylamine in acetonitrile solution at room temperature for 1 hour and then at 0 ° C. To produce a deprotected pyrrolidine compound (VI),

2) Methanesulfonyl compound (VII) was reacted in acetonitrile solution with potassium thioacetic acid salt at a reaction temperature of 90 ° C. or higher for 10 hours to form an acetylthioester compound (VII), which was then treated with trifluoroacetic acid and Reacting with oxybenzene for 30 minutes at room temperature to produce a deprotected carboxythioacetyl compound (V),

3) The carboxythioacetyl compound (V) and the pyrrolidine compound (VI) are reacted in an acetonitrile solution with N, N-carbonyldiimidazole at room temperature for 3 hours to form a thioacetyl compound (IV). Steps, and

4) The thioacetyl compound (IV) is reacted for 1 hour at 0 ° C. using an aqueous sodium hydroxide solution as a base in a methanol solution to produce a hydrolyzed thiol compound (III).

The present invention also provides a pharmaceutical composition for antibiotics containing 1-betamethylcarbapenem derivative of formula 1 as an active ingredient.

The 1-betamethylcarbapenem derivative of the present invention measured the minimum bacterial inhibition concentration (MIC, ㎍ / ml), compared with imipenem (merpenem) and mertapenem (ertapenem) used as antibiotics It shows significant antibacterial activity.

In addition, the 1-betamethylcarbapenem derivative of the present invention exhibits excellent antimicrobial activity against both Gram-positive bacteria and Gram-negative bacteria including ESBL and MDR producing strains, and is stable against DHP-I.

The present invention provides, in addition to non-toxic, inert, pharmaceutically suitable excipients, pharmaceutical compositions consisting of one or more compounds according to the invention and methods of preparing said compositions.

The 1-betamethylcarbapenem derivative of the present invention can be administered parenterally during clinical administration and can be used in the form of general pharmaceutical preparations.

That is, the 1-betamethylcarbapenem derivative of the present invention can be administered in various parenteral formulations during actual clinical administration, and the preparation for parenteral administration includes sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, and lyophilized agents. I include As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. To formulate into a parenteral formulation, the 1-betamethylcarbapenem derivative is mixed in water with a stabilizer or buffer to prepare a solution or suspension, which is formulated in unit dosage forms of ampoules or vials.

In general, in the pharmaceutical product, the effective dose of the 1-betamethylcarbapenem derivative according to the present invention is 0.1 to 100 mg / kg, preferably 0.1 to 10 mg / kg, and may be administered once to several times a day. . However, the dosage may need to be changed, and in particular, may be changed in consideration of the constitution specificity and weight of the subject to be treated, the type and depth of the disease, the nature of the formulation, the nature of the drug administration, and the duration or interval of administration.

As a result of the parenteral administration of the 1-betamethylcarbapenem derivative of the present invention to rats, 50% lethal dose (LD ₅₀ ) of the 1-betamethylcarbapenem derivative was found to be 2 g / kg or more, which is considered a safe substance. do.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.

Example 1  : 1-N- (tert-butoxycarbonyl) -3-hydroxyiminopyrrolidine

9.3 g (50 mmol) of 1-N- (tert-butoxycarbonyl) -3-oxopyrrolidine was dissolved in 80 ml of ethanol and 40 ml of tetrahydrofuran, followed by hydroxylamine hydrochloride (XI) 5.2 g (75 mmol) and 6.3 g (75 mmol) of sodium bicarbonate were dissolved in 40 ml of distilled water, and added dropwise, followed by stirring at room temperature for 1 hour. After distillation under reduced pressure to remove the solvent, the residue was purified by recrystallization with ethyl acetate and hexane to obtain 9.0 g (90%) of 1-N- (tert-butoxycarbonyl) -3-hydroxyiminopyrrolidine (Ⅶ). Obtained in a white solid state.

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.49 (s, 9H), 2.70 (t, 2H, J = 7.4 Hz), 2.80 (t, 2H, J = 7.4 Hz), 3.60-3.62 (m, 2H) , 4.05 (s, 2H), 4.15 (s, 2H), 8.52 (bs, 1H).

Example 2  : (3S, 5S) -3-acetylthio-5-paramethoxybenzyloxycarbonyl-1-allyloxy carbonylpyrrolidine

27.0 g (65.3 mmol) of (3S, 5S) -3-methanesulfonyloxy-5-paramethoxybenzyloxycarbonyl-1-allyloxycarbonylpyrrolidine was dissolved in 400 ml of acetonitrile. Then, 14.9 g (0.13 mol) of potassium thioacetate salt was added thereto, and the mixture was heated to reflux at 90 ° C or higher. The solvent was removed by distillation under reduced pressure, and extracted with water and ethyl acetate. The organic solvent layer was dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent. The residue was purified by column chromatography to give 23.1 g (90%) of (3S, 5S) -3-acetylthio-5-paramethoxybenzyloxycarbonyl-1-allyloxycarbonylpyrrolidine. Obtained as a brown liquid.

¹ H NMR (300MHz, CDCl ₃ ) δ 2.14-2.34 (m, 1H), 2.29 (s, 3H), 2.55-2.71 (m, 1H), 3.80 (s, 3H), 3.86-3.91 (m, 1H ), 4.48-4.50 (m, 2H), 4.60-4.63 (m, 1H), 5.05- 5.10 (m, 2H), 5.13-5.24 (m, 2H), 5.70-5.95 (m, 1H), 6.88 (d , 2H, J = 12.0 Hz), 7.27 (d, 2H, J = 12.0 Hz).

Example 3  : (3S, 5S) -3-acetylthio-5-carboxy-1-allyloxycarbonylpyrrolidine (V)

20.0 g (51 mmol) of (3S, 5S) -3-acetylthio-5-paramethoxybenzyloxycarbonyl-1-allyloxycarbonylpyrrolidine was dissolved in 80 ml of trifluoroacetic acid, 11.1 mL (102 mmol) of methoxybenzene was added dropwise, followed by stirring at room temperature for 30 minutes. The solvent was removed by distillation under reduced pressure, and extracted with water and ethyl acetate. The organic solvent layer was dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent. The residue was purified by column chromatography to obtain 10.0 g (72%) of (3S, 5S) -3-acetylthio-5-carboxy-1-allyloxycarbonylpyrrolidine (V) as a pale brown liquid.

¹ H NMR (300MHz, CDCl ₃ ) δ 2.02-2.11 (m, 1H), 2.31 (s, 3H), 2.70-2.77 (m, 1H), 3.35 (m, 1H), 3.93-4.08 (m, 2H ), 4.40-4.45 (m, 1H), 4.59 (m, 2H), 5.18-5.88 (m, 2H), 5.92-5.99 (m, 2H), 9.90 (bs, 1H).

Example 4  : (3S, 5S) -3-acetylthio-5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine (IV)

After dissolving 1.8 g (8.4 mmol) of 1-N- (tert-butoxycarbonyl) -3-hydroxyiminopyrrolidine in 30 ml of acetonitrile, 13 ml (0.17 mol) of trifluoroacetic acid Was slowly added dropwise at 0 ° C. and then reacted at room temperature for 1 hour, followed by distillation under reduced pressure to remove the solvent. The obtained deprotected 3-hydroxyiminopyrrolidine (VI) was dissolved in 10 ml of acetonitrile, and 3.5 ml (21.0 mmol) of triethylamine was added dropwise at 0 ° C.

1.9 g (7.0 mmol) of (3S, 5S) -3-acetylthio-5-carboxy-1-allyloxycarbonylpyrrolidine (V) was dissolved in 30 ml of acetonitrile and then N, N-carbodiimide It was activated by adding 1.2 g (7.0 mmol) of doazole. Deprotected 3-hydroxyiminopyrrolidine (VI) was slowly added dropwise thereto, followed by stirring at room temperature for 3 hours. The solvent was removed by distillation under reduced pressure, and extracted with water and ethyl acetate. The organic solvent layer was dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent. The residue was purified by column chromatography to give (3S, 5S) -3-acetylthio-5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine (IV). ) 1.25 g (50%) was obtained as a pale brown liquid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.98-2.09 (m, 1H), 2.34 (s, 3H), 2.62-2.79 (m, 2H), 2.90-2.99 (m, 1H), 3.37-3.4 (t , 1H), 3.63-3.82 (m, 1H), 3.88-4.05 (m, 2H), 4.08-4.16 (m, 2H), 4.28-4.45 (m, 1H), 4.48-4.54 (m, 1H), 4.57 -4.63 (m, 2H), 5.20-5.33 (m, 2H), 5.83-5.98 (m, 1H).

Example 5  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidin-3-yl Thio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid allyl ester (XII)

0.50 g (1.4 mmol) of (3S, 5S) -3-acetylthio-5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine (IV) After dissolving in 20 ml of methanol, 1.68 ml (1.68 mmol) of 1N aqueous sodium hydroxide solution was slowly added dropwise at 0 ° C. After the reaction was completed, neutralized with 1M hydrochloric acid solution and distilled under reduced pressure to remove the solvent. After extraction with water and ethyl acetate, the organic solvent layer was dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent, followed by (3S, 5S) -3-mercapto-5- (3-hydroxyiminopyrroli). Din-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine (III) was obtained.

On the other hand, (1R, 5S, 6S) -6-[(1R) -1-hydroxymethyl] -1-methyl-2-diphenylphosphoryloxycarbafen-2-emcarboxylic acid allyl ester (II) 1.07 g (1.8 mmol) was dissolved in 30 mL of acetonitrile, and 0.47 mL (2.7 mmol) of N, N-diisopropylethylamine was added to an ice bath. Herein, (3S, 5S) -3-mercapto-5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine (III) is added to acetonitrile. The dissolved solution was slowly added dropwise. After stirring for 3 hours at the same temperature, the mixture was extracted with ethyl acetate and saturated aqueous sodium chloride solution. The organic solvent layer was dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent, and then the residue was purified by column chromatography (1R, 5S, 6S) -2-[(3S, 5S) -5- (3- Hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidine-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphene- 0.21 g (25%) of 2-m-3-carboxylic allyl ester (XII) was obtained as a brown liquid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.25 (d, 3H, J = 7.1 Hz), 1.34 (d, 3H, J = 6.1 Hz), 1.98-2.07 (m, 1H), 2.67-2.84 (m, 3H), 3.25-3.52 (m, 3H), 3.57-3.90 (m, 3H), 4.03-4.27 (m, 4H), 4.62-4.82 (m, 6H), 5.20-5.47 (m, 4H), 5.93- 6.06 (m, 2 H).

Example 6  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6- [ (1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid (I)

(1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) -1-allyloxycarbonylpyrrolidin-3-ylthio ] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid allyl ester (XII) 0.14 g (0.25 mmol) was distilled off under a nitrogen stream. After dissolving in 15 ml of methylene chloride and adding catalytic amount of tetrakis (triphenylphosphine) palladium (0) under ice bath, 0.15 ml (0.55 mmol) of tributyltin hydride was added dropwise. After 3 hours, when the reaction was completed, the mixture was extracted with distilled water, the methylene chloride layer was removed, and the distilled water layer was taken and freeze-dried. The lyophilized impure compound was purified by Diaion HP-20 tube chromatography (3% aqueous tetrahydrofuran solution) to give (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino Pyrrolidin-1-ylcarbonyl) pyrrolidin-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid ( I) 50 mg (46%) was obtained as a white solid.

¹ H NMR (300 MHz, D ₂ O) δ 1.09 (d, 3H, J = 7.2 Hz), 1.17 (d, 3H, J = 6.3 Hz), 1.78-1.84 (m, 1H), 2.69-2.76 (m , 2H), 2.79-2.86 (m, 1H), 3.25-3.29 (m, 2H), 3.33-3.36 (m, 1H), 3.48-3.52 (m, 1H), 3.57-3.80 (m, 2H), 3.82 -3.90 (m, 1H), 4.09-4.17 (m, 3H), 4.29-4.35 (m, 1H), 4.43-4.49 (m, 1H);

FABHRMS m / z Calcd for C ₁₉ H ₂₇ N ₄ O ₆ S (M + H) ⁺ 439.1651, Found 439.1649.

Example 7-11  :

Thiol (1R, 5S, 6S) -6-[(1R) -1-hydroxymethyl] -1-methyl-2-diphenylphosphoryloxycarbafen-2-emcarboxylic acid allyl ester compound (II) Carbapenem compound (I) was obtained by reacting compound (III) with Examples 1-6 exemplified above.

The analysis results of these compounds are as follows, and the structure is shown in Table 1.

Example 7  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6-[( 1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

¹ H NMR (300 MHz, D ₂ O) δ 1.10 (d, 3H, J = 7.2 Hz), 1.18 (d, 3H, J = 6.3 Hz), 1.78-1.84 (m, 1H), 2.69-2.76 (m , 2H), 2.79-2.86 (m, 1H), 3.25-3.29 (m, 2H), 3.33-3.36 (m, 1H), 3.48-3.52 (m, 1H), 3.57-3.80 (m, 2H), 3.77 (s, 3H), 3.82-3.90 (m, 1H), 4.09-4.17 (m, 3H), 4.29-4.35 (m, 1H), 4.43-4.49 (m, 1H);

FABHRMS m / z Calcd for C ₂₀ H ₂₉ N ₄ O ₆ S (M + H) ⁺ 453.1808, Found 453.1799.

Example 8  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

¹ H NMR (300 MHz, D ₂ O) δ 1.11-1.13 (d, 3H, J = 7.2 Hz), 1.19-1.21 (d, 3H, J = 6.4 Hz), 1.97 (m, 1H), 2.73 (m , 1H), 3.25-3.39 (m, 3H), 3.36 (m, 2H), 3.51 (m, 2H), 3.55 (m, 2H), 3.76 (m, 1H), 4.07-4.18 (m, 4H);

FABHRMS m / z Calcd for C ₂₀ H ₃₀ N ₅ O ₆ S (M + H) ⁺ 468.1917, Found 468.1918.

Example 9  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

¹ H NMR (300 MHz, D ₂ O) δ 1.08-1.11 (d, 3H, J = 7.2 Hz), 1.17-1.19 (d, 3H, J = 6.4 Hz), 1.80 (m, 1H), 1.82 (m , 1H), 2.75 (m, 1H), 3.11 (m, 2H), 3.31-3.37 (m, 4H), 3.53 (m, 2H), 3.75 (m, 1H), 3.80 (m, 3H), 3.81- 3.93 (m, 2 H), 4.11-4.16 (m, 2 H);

FABHRMS m / z Calcd for C ₂₁ H ₃₂ N ₅ O ₆ S (M + H) ⁺ 482.2073, Found 482.2083.

Example 10  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-carboxypyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio]- 6-[(1R) -1-hydroxyethyl] -1-methylcarbaphen-2-m-3-carboxylic acid

¹ H NMR (300 MHz, D ₂ O) δ 1.12 (d, 3H, J = 7.2 Hz), 1.19 (d, 3H, J = 6.3 Hz), 1.81-1.86 (m, 1H), 2.52-2.93 (m , 3H), 3.25-3.39 (m, 3H), 3.51-3.86 (m, 3H), 3.88-3.96 (m, 1H), 4.07-4.23 (m, 4H);

FABHRMS m / z Calcd for C ₂₀ H ₂₇ N ₄ O ₈ S (M + H) ⁺ 483.1550, Found 483.1537.

Example 11  : (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-ethoxycarbonylpyrrolidin-1-ylcarbonyl) pyrrolidin-3-yl Thio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid

¹ H NMR (300 MHz, D ₂ O) δ 1.05-1.15 (m, 9H), 1.77-1.92 (m, 1H), 2.91-2.97 (m, 1H), 3.21 (t, 1H), 3.30-3.34 ( m, 2H), 3.60-3.64 (m, 1 H), 3.82-4.36 (m, 9H), 4.61-4.68 (m, 2H);

FABHRMS m / z Calcd for C ₂₂ H ₃₁ N ₄ O ₈ S (M + H) ⁺ 511.1863, Found 511.1870.

R ₁ R ₂

Example 6 H H Example 7 CH ₃ H Example 8 H CH ₂ NH ₂ Example 9 CH ₃ CH ₂ NH ₂ Example 10 H CO ₂ H Example 11 H CO ₂ Et

Experimental Example 1  : Antimicrobial Activity Assay and DHP-I Stability Test for Standard Strains

In order to determine the antimicrobial activity of the 1-betamethylcarbapenem derivative of the present invention, the following experiment was performed using the agar dilution method.

Gram-positive bacteria such as Streptococcus and Staphylococcus were used as test strains, and Escherichia coli, Pseudomonas aeruginosa, Salmonella, Klebsiella and Enterobacteriaceae were used as Gram-negative bacteria.

After culturing the test strains in dilute agar medium, the 1-betamethylcarbapenem derivatives of the present invention obtained in Examples 6 to 11 were treated to represent the minimum strain concentration (MIC, ㎍ / ㎖), and the standard strain In vitro activity tests were performed.

Imiphenem and meropenem were used as controls.

Susceptibility experiments on the renal secreted DHP-I enzyme were performed against the same control. This sensitivity test set the _half- life (t _1/2 ) of meropenem to DHP-I enzyme to 1.00 to estimate the relative stability upon hydrolysis of each drug.

The results are shown in Table 2.

Strain Minimum Strain Inhibition Concentration (MIC, ㎍ / ㎖) Example Control 6 7 8 9 10 11 Imipenem Meropenem One Streptococcus. pyogenes 308A 0.013 - 0.002 0.004 0.013 0.013 0.004 0.007 2 Streptococcus. pyogenes 77A 0.007 - 0.07 0.007 0.013 0.007 <0.002 0.007 3 Streptococcus. faecium MD 8b 6.250 - 6.025 3.125 6.250 12.500 0.781 12.500 4 Staphylococcus. aureus SG511 0.195 - 0.098 0.098 0.098 0.195 0.013 0.098 5 Staphylococcus. aureus 285 0.195 - 0.195 0.195 0.195 0.391 0.013 0.195 6 Staphylococcus. aureus 503 0.098 - 0.098 0.049 0.098 0.195 0.007 0.098 7 Escherichia. coli 07 0.025 0.049 0.025 0.025 0.013 0.013 0.098 0.025 8 Escherichia. coli DC 0 0.025 0.049 0.049 0.049 0.025 0.049 0.195 0.025 9 Escherichia. coli DC 2 0.025 0.049 0.049 0.049 0.025 0.025 0.195 0.025 10 Escherichia. coli tem 0.025 0.049 0.049 0.049 0.025 0.025 0.098 0.025 11 Escherichia. coli 1507E 0.025 0.049 0.049 0.049 0.025 0.049 0.098 0.025 12 Pseudomonas. aeruginosa 9027 0.391 0.391 0.195 0.391 0.391 6.250 0.391 0.195 13 Pseudomonas. aeruginosa 1592E 0.391 3.125 0.098 0.195 0.391 3.125 0.781 0.098 14 Pseudomonas. aeruginosa 1771 0.781 6.35 0.195 0.391 0.781 6.250 0.781 0.391 15 Pseudomonas. aeruginosa 1771M 0.195 0.781 0.098 0.098 0.195 0.391 0.195 0.098 16 Salmonella. typhimurium 0.025 0.098 0.098 0.098 0.049 0.049 0.781 0.049 17 Klebsiella. cxytoca 1082E 0.098 0.195 0.195 0.391 0.098 0.391 0.195 0.049 18 Klebsiella. aerogenes 1522E 0.025 0.049 0.098 0.098 0.025 0.049 0.195 0.049 19 Enteroabacter cloacae P99 0.049 0.098 0.195 0.098 0.098 0.781 0.098 0.049 20 Enteroabacter cloacae 1321E 0.013 0.049 0.049 0.025 0.025 0.025 0.098 0.025 DHP-I susceptibility 1.03 - 0.31 0.25 0.54 0.26 0.18 1.00

As shown in Table 2, the 1-betamethylcarbapenem derivatives of the present invention showed an excellent and balanced antimicrobial activity equivalent to that of meropenem with respect to Gram-negative bacteria, including Gram-positive bacteria and Pseudomonas aeruginosa.

In particular, the compounds of Examples 6 and 8 having an oxime group showed better antimicrobial activity against Gram-negative bacteria than the compounds of Examples 7 and 9 having a methyloxime group. The compound of Example 10 having a carboxylic acid group was about twice as good as the Gram-positive bacteria than the compound of Example 11 having a carboxylic acid ester group.

Among the 1-betamethylcarbapenems of the present invention, the compound of Example 6 having a simple oxime group showed a broad and excellent antimicrobial effect equivalent to meropenem. In addition, the stability of the DHP-I enzyme far exceeded that of imipenem, which does not have 1-betamethyl group, and showed somewhat superior stability compared to meropenem, which is a notable substance requiring several more advanced experiments. Judging.

Experimental Example 2  : Antimicrobial Activity Assay for Clinically Isolated Gram-negative Strains

In order to determine the antimicrobial activity of the 1-betamethylcarbapenem derivative of the present invention, the following experiment was performed using the agar dilution method.

Gram-negative E. coli, Pseudomonas aeruginosa, Salmonella, Klebsiella, Enterobacteriaceae and Shigella bacteria including 40 isolated ESBL (extended-spectrum β-lactamase) and MDR (multi-drug resistant) producing strains were tested as test strains. Used.

After culturing the test strains in dilute agar medium, the 1-betamethylcarbapenem derivative of Example 6 was treated to show the minimum strain concentration (MIC, μg / ml), and the in vitro activity test was performed on the standard strain. It was.

Meropenem and Ertapenem were used as controls.

The results are shown in Table 3.

Strain Minimum Strain Inhibition Concentration (MIC, ㎍ / ㎖) Example 6 Meropenem Ultapenem One Escherichia. coli DC0 0.03 0.03 ≤0.008 2 Escherichia. coli DC2 0.016 0.03 0.016 3 Escherichia. coli tem 0.016 0.016 0.016 4 Escherichia. coli ATCC25922 0.016 0.016 ≤0.008 5 Escherichia. coli 0157: H7 0.016 0.03 ≤0.008 6 Escherichia. coli CCARM 1108 MDR 0.03 0.03 0.016 7 Escherichia. coli CCARM 1109 MDR 0.06 0.03 0.06 8 Escherichia. coli CCARM 1110 MDR 0.06 0.016 0.03 9 Escherichia. coli CCARM 1111 MDR 0.016 0.016 ≤0.008 10 Escherichia. coli CCARM 1112 MDR 0.016 0.016 0.016 11 Escherichia. coli CCARM 1113 MDR 0.06 0.03 0.06 12 Escherichia. coli CCARM 1114 MDR 0.13 0.03 0.25 13 Escherichia. coli CCARM 1115 MDR 0.03 0.016 0.06 14 Escherichia. coli CCARM 1116 MDR 0.016 0.03 ≤0.008 15 Escherichia. coli CCARM 1117 MDR 0.03 0.03 0.03 16 Klebsiella pneumoniae 1 ESBL 0.03 0.03 0.016 17 Klebsiella pneumoniae 2 ESBL 0.03 0.03 0.016 18 Klebsiella pneumoniae 3 ESBL 0.13 0.06 0.5 19 Klebsiella pneumoniae 4 ESBL 0.03 0.016 0.06 20 Klebsiella pneumoniae 5 ESBL 0.016 0.016 ≤0.008 21 Klebsiella pneumoniae 6 MDR 0.03 0.03 0.06 22 Klebsiella pneumoniae 7 MDR 0.06 0.06 0.25 23 Klebsiella pneumoniae 8 MDR 0.25 0.06 One 24 Klebsiella pneumoniae 9 MDR 0.03 0.03 0.016 25 Klebsiella pneumoniae 10 MDR 0.016 0.016 0.016 26 Salmonella typhimurium 0.03 0.03 0.016 27 Salmonella typhimurium 1 0.016 0.03 ≤0.008 28 Salmonella typhimurium 2 0.016 0.016 ≤0.008 29 Klebsiella oxytoca 1082E 0.13 0.06 0.03 30 Enterobacter cloacae P99 0.13 0.03 0.06 31 Salmonella enteritidis 0.016 0.016 ≤0.008 32 Salmonella enteritidis 1 0.016 0.016 ≤0.008 33 Salmonella enteritidis 2 ≤0.008 0.016 ≤0.008 34 Salmonella enteritidis 3 0.016 0.03 ≤0.008 35 Shigella flexneri ATCC9199 0.016 0.03 ≤0.008 36 Salmonella sonnei ATCC9290 0.016 0.03 ≤0.008 37 Pseudomonas aeruginosa ATCC27853 One 0.5 4 38 Pseudomonas aeruginosa 1 MDR One One 8 39 Pseudomonas aeruginosa 2 MDR 8 8 8 40 Pseudomonas aeruginosa 3 MDR 8 8 8

As shown in Table 3, the 1-betamethylcarbapenem derivative of Example 6 exhibited excellent antimicrobial activity against 40 target Gram-negative bacteria except MDR Pseudomonas aeruginosa, almost equivalent to meropenem and superior to Ultapenem. Activity was shown. The compound of Example 6 was about 2 to 4 times better than that of Ultafenem with respect to MDR Krebs.

Therefore, it can be seen that the compound of Example 6 exhibits excellent antimicrobial activity against Gram-negative bacteria including ESBL and MDR producing strains.

Experimental Example 3  : Acute Toxicity in Parenteral Administration of Rats

In order to determine the acute toxicity of the 1-betamethylcarbapenem derivative of the present invention, the following experiment was performed.

Acute toxicity test was performed using 6-week-old SPF SD rats. The compound of Example 6 was suspended in 1 ml of saline and administered intravenously to the tails of the two rats at doses of 1000 mg / kg and 2000 mg / kg.

After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed, and hematological and hematological examinations were performed.

As a result, there were no clinical symptoms or deaths in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemistry test, autopsy findings, etc.

As a result, the 1-betamethylcarbapenem derivative of the present invention did not show toxicity change up to 2000 mg / kg in rats, and the parenteral administration minimum dose (LD ₅₀ ) was determined to be a safe substance of 2 g / kg or more.

Since the 1-betamethylcarbapenem derivative of the present invention shows excellent antimicrobial activity against Gram-negative bacteria including not only Gram-positive bacteria but also ESBL (extended-spectrum β-lactamase) and MDR (multi-drug resistant) producing strains, It can be usefully used.

Claims (13)

1-betamethylcarbapenem derivatives represented by the following formula (1) and their pharmaceutically acceptable salts. <Formula 1>

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.) The 1-betamethylcarbapenem derivative and pharmaceutically acceptable salts thereof according to claim 1, wherein R ₁ is hydrogen and R ₂ is hydrogen, aminomethyl or carboxy. The method of claim 1, wherein the 1-betamethylcarbapenem derivative is 1) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6- [(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid, 2) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyiminopyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6- [ (1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid, 3) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio ] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid, 4) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-methoxyimino-4-aminomethylpyrrolidin-1-ylcarbonyl) pyrrolidin-3-ylthio ] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid, 5) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-carboxypyrrolidin-1-ylcarbonyl) pyrrolidine-3-ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid and 6) (1R, 5S, 6S) -2-[(3S, 5S) -5- (3-hydroxyimino-4-ethoxycarbonylpyrrolidin-1-ylcarbonyl) pyrrolidine-3- 1-betamethylcarbapenem, which is selected from the group consisting of ylthio] -6-[(1R) -1-hydroxyethyl] -1-methylcarbafen-2-m-3-carboxylic acid Derivatives and their pharmaceutically acceptable salts. 1) preparing a carbapenem derivative (XII) by reacting a carbapenem enol phosphate compound (II) with a thiol derivative (III) in the presence of a base (first step), and 2) deprotecting the carbapenem derivative (XII) obtained in the first step to prepare 1-betamethylcarbapenem derivative (I) (second step), A method for preparing the 1-betamethylcarbapenem derivatives of claim 1 represented by Scheme 1 and their pharmaceutically acceptable salts. <Scheme 1>

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.) The method of claim 1, wherein the base in the first step is N, N-diisopropylethylamine, characterized in that the 1-betamethyl carbapenem derivatives and pharmaceutically acceptable salts thereof. The 1-betamethylcarbapenem derivative of claim 1 and the pharmaceutically acceptable compound of claim 1, wherein the reaction solvent in the first step is acetonitrile and the reaction conditions are reacted at 0 ° C. for 3 hours. Preparation of Salts. The method of claim 4, wherein the reaction solvent in the second step is methylene chloride, the catalyst is tributyl tin hydride and tetrakis (triphenylphosphine) palladium (0), the reaction conditions are reacted for 2 hours at 0 ℃ A method for producing the 1-betamethylcarbapenem derivatives of claim 1 and their pharmaceutically acceptable salts. Thiol derivative (III) represented by the following general formula (2) used to prepare the 1-betamethylcarbapenem derivative of claim 1 and pharmaceutically acceptable salts thereof. <Formula 2>

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.) 1) A pyrrolidinone compound (X) reacted with a butoxycarbonyl group-protected pyrrolidinone compound (X) and a hydroxylamine or methoxyamine compound (XI) in the presence of a base to produce a pyrrolidine compound (X), which undergoes a deprotection reaction. Producing pyrrolidin compound (VI) through (first step), 2) reacting the methanesulfonyl compound (VII) with potassium thioacetic acid salt in an acetonitrile solution to produce an acetylthioester compound (VII), and producing a carboxythioacetyl compound (V) through a deprotection reaction ( Step 2), 3) reacting the carboxythioacetyl compound (V) obtained in the second step with the pyrrolidine compound (VI) obtained in the step 1) to produce a thioacetyl compound (IV) (third step), and 4) hydrolyzing the thioacetyl compound (IV) to produce a thiol compound (III) (fourth step), A thiol derivative (III) of claim 8 represented by Scheme 2 and a method for preparing a pharmaceutically acceptable salt thereof. <Scheme 2>

(R ₁ is hydrogen or methyl group, R ₂ is hydrogen, aminomethyl, carboxy or ethoxycarbonyl group.) 10. The method of claim 9, wherein the reaction solvent in the first step is a mixed solution of ethanol and tetrahydrofuran (2: 1), the base is sodium hydrogen carbonate, the reaction conditions are characterized in that for 1 hour at 0 ~ 40 ℃ A thiol derivative (III) of claim 8 and a method for preparing a pharmaceutically acceptable salt thereof. 10. The method for preparing thiol derivative (III) of claim 8 and their pharmaceutically acceptable salts according to claim 9, wherein the reaction conditions in the second step are reacted at 90 DEG C for 10 hours. 10. The thiol derivative (III) of claim 8 and a pharmaceutical composition thereof according to claim 9, characterized in that the reaction is carried out at room temperature for 3 hours using N, N-carbonyldiimidazole in an acetonitrile solution in the third step. Method for the preparation of salts that are acceptable. The pharmaceutical composition for antibiotics containing the 1-betamethylcarbapenem derivative of claim 1 and pharmaceutically acceptable salts thereof as an active ingredient.