KR100523986B1 - Carbapenem derivatives with an antibacterial activity against resistant strains and preparation thereof - Google Patents

Abstract

The present invention relates to a carbapenem derivative represented by the following Chemical Formula 1 having an antibiotic action against resistant bacteria, including Methicillin Resistant Staphylococcus Aureus (MRSA), a preparation method thereof, and an antibiotic composition containing the same. Since the penem derivative is very excellent in antibacterial effect, it can be usefully used as an antibiotic against infection of intractable resistant bacteria including MRSA and oploxacin resistant strain (QRSA).

Where

X represents a sulfur or nitrogen atom;

Y represents a hydrogen, nitro group or amine group present at a specific position of the phenyl group;

M represents a hydrogen ion or a counterion forming a pharmacologically acceptable salt.

Description

Carbapenem derivatives having an antibiotic action against resistant bacteria and a method for preparing the same {CARBAPENEM DERIVATIVES WITH AN ANTIBACTERIAL ACTIVITY AGAINST RESISTANT STRAINS AND PREPARATION THEREOF}

The present invention relates to a carbapenem derivative having an antibiotic action against resistant bacteria including Methicillin Resistant Staphylococcus Aureus (MRSA), a preparation method thereof, and an antibiotic composition containing the same.

First used in the clinic as a modern concept antibiotic in the 1940s, penicillin has been known as a miracle medicine by saving the lives of many patients with infectious diseases. However, soon after penicillin was used, staphylococci appeared resistant. In the 1960s, methicillin, a semisynthetic penicillin, was developed and used to treat penicillin-resistant staphylococcal infections. In 1973, cephalosporin-based cefazolines were developed. However, methicillin-resistant staphylococcus aureus (MRSA), which is resistant to methicillin or cefazoline, began to develop, causing another problem. Subsequently, in the 1980s, various cephalosporin preparations and many antibiotics such as quinolone, carbapenem, monobactam, and glycoside were developed and used in actual clinical practice. However, at this time, penicillin-resistant pneumococci, MRSA, etc. began to be a problem all over the world, and in the 1990s, the emergence of vancomycin-resistant aureus (VRE), the problem of antibiotic-resistant bacteria has emerged as a concern of the world pharmaceutical community.

In 1996, vancomycin-resistant Staphylococcus aureus (VRSA) appeared in Japan for the first time, and the efficacy of the glycoside preparation, considered the last bulwark, began to shake significantly, and the frequency of Gram-negative bacteria producing a wide range of beta-lactamase (ESBL) also began. It started to increase.

Recently, 2-arylcarbapenem compounds (MER-6), a carbapenem antibiotic, (L-695256 and L-742728) have been reported to show good activity against MRSA and VRSA (Hugh rosen et al., Sciences , 703). (1999), International Publication No. WO 99/62906 has reported that 2-benzothiazoleethenyl carbapenem shows good effect on MRSA. In addition, a number of carbapenem antibiotics that have a good effect on MRSA have been reported, and among these, imipenem and meropenem are used for the treatment of MRSA with weak resistance.

The problem of antibiotic resistance inevitably results in failure of patient treatment, and the development of new antibiotics to overcome this situation is strongly required.

Accordingly, the present inventors continued to develop antibiotics having a therapeutic effect against infection of resistant bacteria, including MRSA, and thus, the present invention was completed by developing a new carbapenem derivative.

Accordingly, an object of the present invention is to provide a carbapenem derivative substituted with 2-benzothiazolo pyrrolidine or 2-benzimidazolo pyrrolidine derivative having a antibiotic action against a resistant bacterium and a preparation method thereof. It is.

Another object of the present invention is to provide an antibiotic composition containing the carbapenem derivative as an active ingredient.

In order to achieve the above object, the present invention provides a carbapenem derivative of Formula 1 or a pharmacologically acceptable salt thereof.

Formula 1

Where

X represents a sulfur or nitrogen atom;

Y represents a hydrogen, nitro group or amine group present at a specific position of the phenyl group;

M represents a hydrogen ion or a counterion forming a pharmacologically acceptable salt.

In addition, in the present invention, a compound of formula 2 is reacted with a compound of formula 3 to prepare a carbapenem ester derivative of formula 4, and then a carboxy protecting group, an amine protecting group and optionally a hydroxy protecting group are removed from the compound of formula 4. Provided is a method of preparing a carbapenem derivative of Formula 1, comprising:

Wherein R ₁ is a hydrogen atom or a hydroxy protecting group such as tert-butyldimethylsilyl group or triethylsilyl group;

R ₂ is a carboxy protecting group such as p-nitrobenzyl group, allyl group or p-methoxybenzyl group;

R ₃ is an amine protecting group such as p-nitrobenzyloxycarbonyl group, p-methoxybenzyloxycarbonyl group or allyloxycarbonyl group;

X 'is a sulfur atom or a nitrogen atom protected with an amine protecting group such as p-nitrobenzyloxycarbonyl group, p-methoxybenzyloxycarbonyl group or allyloxycarbonyl group;

Y is as defined above.

In order to achieve the above another object, the present invention provides an antibiotic composition comprising an effective amount of a carbapenem derivative of Formula 1 or a pharmacologically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Hereinafter, the present invention will be described in more detail.

Carbapenem derivatives of the general formula (1) of the present invention, in particular,

Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1,3-benzothiazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6- [ (1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate,

Sodium (4S, 6R) -3-({(3S, 5R) -5-[(4-amino-1,3-benzothiazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6-[(1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate,

Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1H-benzimidazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6-[(1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, and

Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1H-4-nitrobenzimidazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6- [(1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate and the like are preferable.

In the present invention, the carbapenem derivative of Formula ₁ is prepared by reacting a compound of Formula 2 with a compound of Formula 3 to prepare a compound of Formula 4, and then when R ₁ is a hydrogen atom, R ₂ is a carboxy protecting group in the compound of Formula 4 the amine protecting group of R ₃ a may be prepared by removing at the same time, R ₁ is hydroxy protective group in case a the R ₃ hydroxy protecting group R _1, a carboxy protecting group R ₂ and an amine protecting group in the compound of formula (4) at the same time or It can manufacture by removing in order.

Specifically, the compound of formula 3 is dissolved in a dried organic solvent (e.g. acetonitrile, dimethylformamide or acetone, preferably acetonitrile), cooled to -30 to -20 ° C, and then an organic base (e.g. : N, N-diisopropylethylamine) was added slowly. Subsequently, a compound of Formula 2 may be added thereto and reacted at −5 to 5 ° C. for 2 to 4 hours to prepare a compound of Formula 4.

Removal of the carboxy protecting group (R ₂ ) and the amine protecting group (R ₃ ) in the compound of the formula (4) thus obtained may be carried out simultaneously according to a conventional method. For example, when R ₂ is a p-nitrobenzyl group or p-methoxybenzyl group and R ₃ is a p-nitrobenzyloxycarbonyl group or p-methoxybenzyloxycarbonyl group, it is reduced by hydrogenation catalysis and simultaneously removed. Triphenylphosphine in the presence of an organic acid or salt thereof (e.g. acetic acid, 2-ethylhexanoic acid, or sodium or potassium salt thereof) when R ₂ is an allyl group and R ₃ is an allyloxycarbonyl group. Or by reacting with tetrakis (triphenylphosphine) palladium (0) at the same time.

In addition, in the formula (3), a compound wherein X 'is a nitrogen atom protected with an amine protecting group is prepared by substituting an amine protecting group of the same kind as R ₃ with a nitrogen atom, and the protecting group is simultaneously removed under the condition of removing the R ₂ protecting group in formula (4). can do.

When R ₁ is a hydroxy protecting group, the removal of the hydroxy protecting group in the compound of formula 4 may also be carried out according to conventional methods. For example, when R ₁ is a tert-butyldimethylsilyl group, the compound of formula 4 may be removed by reaction with ammonium bifluoride at room temperature for about 3 days in an N-methylpyrrolidinone solvent, and R ₁ is In the case of the triethylsilyl group, the compound of formula 4 may be removed by reaction with tetrabutylammonium fluoride (TBAF) at room temperature for about 5 hours in a tetrahydrofuran solvent.

The reaction product obtained after the removal of the protecting group may be purified by reverse phase C18-silica gel column chromatography to obtain a compound of formula 1 which is an end product in an amorphous solid state.

According to the present invention, the compound of formula (2) used to prepare the compound of formula (1) can be easily prepared by conventional methods, the compound of formula (3) can be prepared according to the method shown in Scheme 1 below.

Wherein R ₃ , X, X 'and Y are as defined above.

Looking at the preparation method of the compound of formula 3 according to the reaction scheme 1 in detail as follows; i) The starting compound of formula 5 was prepared in five steps from L-proline methylester using known methods ( Bioorg. & Med. Chem . 5: 2069-2087, 1997). Together under heating in an acetone solvent for 40 to 48 hours to prepare the methyl iodide compound of formula 6 in quantitative yield, ii) in an organic solvent (e.g. acetone, tetrahydrofuran, dimethylformamide, preferably acetone) Compound 8 is prepared by heating to reflux for 3 to 5 hours with potassium carbonate and thiol compound of formula 7. In the case where X is a nitrogen atom in the compound of formula 8, to protect it, the compound of formula 8 is added to triethylamine and allyloxycarbonyl chloride (or p-methoxybenzyloxycarbonyl chloride or p-nitrobenzyl) in methylene chloride solvent. Oxycarbonyl chloride) was reacted at 0 ° C. for 2 hours to prepare a compound in which the nitrogen atom was protected with an allyloxycarbonyl group (or p-methoxybenzyloxycarbonyl group or p-nitrotrobenzyloxycarbonyl group). I use it. iii) The reaction of removing the hydroxy protecting group of the compound of Formula 8 thus prepared, reacting the compound of Formula 8 with tetrabutylammonium fluoride in a tetrahydrofuran solvent at room temperature for 18 hours to give quantitative yield Iv) the hydroxy group of the compound of formula 9 can be converted to a thioacetyl group by a known mitznobu reaction to prepare a compound of formula 10, in the present invention in an organic solvent (eg tetrahydrofuran) The compound of formula 9 is reacted with triphenylphosphine and diethylazodicarboxylic acid (DEAD) (or diisopropylazocarboxylic acid) at -5 to 5 ° C. for 30 minutes to 2 hours, and then thioacetic acid is added at room temperature. Reacting for about 3 hours to prepare a compound of Formula 10, and v) reacting a compound of Formula 10 with a base such as sodium hydroxide By hydrolysis can be prepared a compound of formula (3).

Compounds of Formula 1 of the present invention are Gram-positive microorganisms such as methicillin resistant strain (MRSA), oploxacin resistant strain (QRSA), penicillin resistant strain (PRSA), enterococcus and Pneumococcus Has good antibacterial activity against.

Accordingly, the present invention provides an antibiotic composition comprising an effective amount of a compound of Formula 1 or a pharmacologically acceptable salt thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition of the present invention may contain the compound of formula 1 as an active ingredient in an amount of 0.1 to 75% by weight, preferably 1 to 50% by weight, based on the total weight of the composition.

The pharmaceutical compositions of the invention can be formulated in a variety of oral or non-oral dosage forms. Formulations for oral administration include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, and granules. These formulations may contain diluents (e.g., lactose, dextrose, water, etc.) in addition to the active ingredients. Cross, mannitol, sorbitol, cellulose and / or glycine), lubricants such as silica, talc, stearic acid and its magnesium or potassium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally starch, agar, alginic acid or Disintegrating or boiling mixtures such as sodium salts thereof and / or absorbents, colorants, flavoring agents, and sweetening agents. The formulations may be prepared by conventional mixing, granulating or coating methods. Also representative of non-oral dosage forms are injectable formulations, preferably isotonic solutions or suspensions.

The composition may contain sterile and / or auxiliaries such as preservatives, stabilizers, hydrating or emulsifying accelerators, salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, which are conventional methods of mixing, granulating Or according to a coating method.

As an active ingredient, the compound of formula 1 may be divided or divided once a day in an amount of 2.5 to 100 mg / kg body weight, preferably 5 to 60 mg / kg body weight, for mammals including humans. Administration can be via oral or parenteral routes.

Hereinafter, the present invention will be described in more detail by the following Preparation Examples and Examples. However, the following Preparation Examples and Examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Preparation Example 1 Allyl (2R, 4R) -tert-Butyldimethylsilyloxy-2- (iodomethyl) -1-pyrrolidine-1-carboxylic acid ester (Compound 6)

Allyl (2R, 4R) -tert-butyldimethylsilyloxy-2- (methanesulfonylmethyl) -1-pyrrolidine-1-carboxylic acid ester (700 mg, 1.78 mmol) was dissolved in 8 ml of acetone and sodium iodide (347 mg, 2.3 Mmol) was heated to reflux for 48 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, the solvent was concentrated under reduced pressure, ethyl ether and water were added to the reaction solution, and the organic layer was separated. The organic layer was washed with aqueous sodium thiosulfate solution and brine, and dried over anhydrous magnesium sulfate. After concentration under reduced pressure, the residue was purified by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 1) to obtain the target compound in quantitative yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.8-1.85 (m, 1H), 2.0-2.2 (m, 2H), 3.3-3.4 (m, 2H), 3.5-3.6 (t, J = 6.4, 16.6 Hz, 1H) 3.8-4.0 (m, 1H), 4.3 (s, 1H) 4.5-4.6 (m, 2H), 5.1-5.3 (m, 2H), 5.8- 6.0 (m, 1 H). ; MS (EI, 70 eV) m / z 425 (M ⁺ ).

Preparation Example 2 Allyl (2R, 4R) -2-{[(1,3-benzothiazol-2-yl) sulfanyl] methyl} -tert-butyldimethyl silyloxy-1-pyrrolidine-1-carboxylic acid Ester (Compound of Formula 8, X '= S, Y = H)

Allyl (2R, 4R) -tert-butyldimethylsilyloxy-2- (iodomethyl) -1-pyrrolidine-1-carboxylic acid ester (compound of Preparation Example 1) (850 mg, 2.00 mmol) was dissolved in 17 mL of acetone. 2-mergatobenzothiazole (330 mg, 2.00 mmol) and potassium carbonate (300 mg, 2.2 mmol) were added and then heated to reflux for 3 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, the solvent was removed under reduced pressure, ethyl acetate and water were added to the reaction solution, the organic layer was separated, washed with saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. Purification by chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 2: 1) afforded the target compound in a yield of 72%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.5-1.7 (m, 1H), 2.0-2.2 (m, 2H), 3.4-3.5 (m, 2H), 3.8-4.0 (m, 1H), 4.3-4.4 (q, J = 6.2, 14.7 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H) , 7.2-7.4 (m, 2H), 7.7-7.9 (dd, J = 12.6, 19.6 Hz, 2H). ; MS (EI, 70 eV) m / z 464 (M ⁺ ).

Preparation Example 3: Allyl (2R, 4R) -2-{[4-amino- (1,3-benzothiazol-2-yl) sulfanyl] methyl} -tert-butyldimethylsilyloxy-1-pyrrolidine -1-carboxylic acid ester (compound of formula 8, X '= S, Y = 4-amino)

Allyl (2R, 4R) -tert-butyldimethylsilyloxy-2- (iodomethyl) -1-pyrrolidine-1-carboxylic acid ester (compound of Preparation Example 1) (100 mg, 0.24 mmol) was added to 5 mL of acetone. After dissolving, 6-amino-2-mergatobenzothiazole (44 mg, 0.24 mmol) and potassium carbonate (36 mg, 0.26 mmol) were added and the mixture was heated to reflux for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, the solvent was removed under reduced pressure, ethyl acetate and water were added to the reaction solution, the organic layer was separated, the organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure, and then column chromatography (column: Purification with silica gel, mobile phase: normal hexane / ethyl acetate = 1: 2) afforded the desired compound in a yield of 81%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.2-1.3 (m, 1H), 2.0-2.2 (m, 2H), 3.4-3.5 (m, 2H), 3.6-3.8 (m, 1H), 4.3-4.4 (q, J = 7.2, 12.4 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H) , 6.8-6.9 (dd, J = 12.6, 19.6 Hz, 1H), 6.9-7.0 (m, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H). ; MS (EI, 70 eV) m / z 479 (M ⁺ ), 422, 298, 284, 240, 196, 182, 156, 142, 108, 73, 59.

Preparation Example 4 Allyl (2R, 4R) -2-[(1H-benzimidazol-2-ylsulfanyl) methyl] -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid ester Of compounds, X '= N, Y = H)

Allyl (2R, 4R) -tert-butyldimethylsilyloxy-2- (iodomethyl) -1-pyrrolidine-1-carboxylic acid ester (compound of Preparation Example 1) (1.8 g, 4.24 mmol) was added to 30 mL of acetone. After dissolving and adding 2-mergatobenzimidazole (530 mg, 3.53 mmol) and potassium carbonate (536 mg, 3.88 mmol), the mixture was heated and stirred for 3 hours. After the same process as in Preparation Example 3 to obtain the target compound in 99% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.8-1.9 (m, 1H), 2.0-2.2 (m, 2H), 2.8-3.0 (m, 2H), 3.4-3.5 (dd, J = 6.4, 12.8 Hz, 2H), 3.5-3.6 (d, J = 6.2 Hz, 1H) 4.5-4.6 (m, 2H), 5.2-5.4 (m, 2H), 5.8-6.0 (m, 1 H), 7.1-7.2 (m, 2 H), 7.4-7.6 (m, 2 H). ; MS (EI, 70 eV) m / z 447 (M ⁺ ), 390, 346, 298, 284, 240, 200, 150, 108, 88, 75, 61.

Preparation Example 5 Allyl (2R, 4R) -2-[(4-nitro-1H-benzimidazol-2-ylsulfanyl) methyl] -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid Esters (compound of formula 8, X '= N, Y = 4-nitro)

Allyl (2R, 4R) -tert-butyldimethylsilyloxy-2- (iodomethyl) -1-pyrrolidine-1-carboxylic acid ester (compound of Preparation Example 1) (1.8 g, 4.24 mmol) was added to 30 mL of acetone. After dissolving and adding 2-mercapto-5-nitrobenzimidazole (530 mg, 3.53 mmol) and potassium carbonate (536 mg, 3.88 mmol), the mixture was heated and stirred for 3 hours. After the same process as in Preparation Example 3 to obtain the target compound in 99% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.8-2.0 (m, 1H), 2.2-2.4 (m, 2H), 3.1 (s, 2H), 3.6- 3.8 (m, 1H), 4.1-4.3 (m, 2H) 4.5-4.6 (m, 2H), 5.1-5.3 (q, J = 7.2, 12.4 Hz, 2H), 5.8-6.1 (m, 1H), 7.2 -7.4 (dd, J = 6.8, 14.7 Hz, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H), 8.1-8.2 (d, J = 6.6 Hz, 1H). ; MS (EI, 70 eV) m / z 492 (M ⁺ ), 435, 391, 329, 298, 284, 240, 196, 156, 142, 108, 86, 75, 59.

Preparation 6: allyl 2 - [({(2R, 4R) -1 - [( allyloxy) carbonyl] -4- tert-butyldimethylsilyloxy-pyrrolidinyl} methyl) sulfanyl] -1H- benzimidazole Dazole-1-carboxylic acid ester (compound of formula 8, X '= N-allyloxycarbonyl, Y = H)

Allyl (2R, 4R) -2-[(1H-benzimidazol-2-ylsulfanyl) methyl] -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid ester (compound of Preparation Example 4) (3.53 mmol) was dissolved in 15 mL of dichloromethane, triethylamine (1.48 mL, 10.59 mmol) was added, and allyloxycarbonyl chloride (0.45 mL, 4.24 mmol) was slowly added dropwise at 0 ° C. After stirring for 2 hours, the solid was removed by filtration and the solvent was concentrated. Ethyl acetate and water were added to the reaction mixture, and the organic layer was separated. The organic layer was washed with a saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to obtain a yellow liquid target compound in a yield of 80%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.8-1.9 (m, 1H), 2.0-2.2 (m, 2H), 2.8-3.0 (m, 2H), 3.4-3.5 (dd, J = 6.4, 12.8 Hz, 2H), 3.5-3.6 (d, J = 6.2 Hz, 1H) 4.5-4.6 (m, 4H), 5.2-5.4 (m, 4H), 5.8-6.0 (m, 2H), 7.1-7.2 (m, 2H), 7.5-7.7 (dd, J = 1.2, 6.0 Hz, 1H), 7.8-7.9 (dd, J = 1.8, 7.6 Hz, 1H). ; MS (EI, 70 eV) m / z 531 (M ⁺ ), 516, 474, 430, 390, 329, 298, 284, 234, 196, 175, 156, 142, 108, 73

Preparation 7: allyl 2 - [({(2R, 4R) -1 - [( allyloxy) carbonyl] -4-tert-butyldimethylsilyloxy-pyrrolidinyl} methyl) sulfanyl] -4-nitro- 1H-benzimidazole-1-carboxylic acid ester (compound of Formula 8, X '= N-allyloxycarbonyl, Y = 4-nitro)

Allyl (2R, 4R) -2-[(4-nitro-1H-benzimidazol-2-ylsulfanyl) methyl] -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid ester (Preparation Example 5 compound) (3.53 mmol) was dissolved in 15 mL of dichloromethane, triethylamine (1.48 mL, 10.59 mmol) was added, and allyloxycarbonyl chloride (0.45 mL, 4.24 mmol) was slowly added dropwise at 0 ° C. After stirring for 2 hours, the solid was removed by filtration and the solvent was concentrated. Ethyl acetate and water were added to the reaction mixture, and the organic layer was separated. The organic layer was washed with saturated aqueous sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to obtain a yellow liquid target compound in a yield of 86%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 0.05 (s, 6H) 0.8 (s, 9H) 1.8-2.0 (m, 1H), 2.2-2.4 (m, 2H), 3.1 (s, 2H), 3.6- 3.8 (m, 1H), 4.1-4.3 (m, 2H) 4.5-4.6 (m, 4H), 5.1-5.3 (q, J = 7.2, 12.4 Hz, 4H), 5.8-6.1 (m, 2H), 7.2 -7.4 (dd, J = 6.8, 14.7 Hz, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H), 8.1-8.2 (dd, J = 6.6, 12.8 Hz, 1H). ; MS (EI, 70 eV) m / z 576 (M ⁺ ), 561, 519, 475, 435, 377, 329, 298, 284, 240, 196, 156, 142, 108, 73.

Preparation Example 8: Allyl (2R, 4R) -2-{[(1,3-benzothiazol-2-yl) sulfanyl] methyl} -4-hydroxy-1-pyrrolidine-1-carboxylic acid ester ( Compound of Formula 9, X '= S, Y = H)

Allyl (2R, 4R) -2-{[(1,3-benzothiazol-2-yl) sulfanyl] methyl} -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid ester 2 compound) (660 mg, 1.42 mmol) in 8.5 ml of tetrahydrofuran and acetic acid (0.85 ml, 14.2 mmol) and tetrahydroammonium fluoride (5.68 ml, 5.68 mmol / 1 mol tetrahydrofuran solution) at 0 ° C. After the addition, the temperature was raised to room temperature, followed by stirring for 18 hours. Saturated sodium bicarbonate solution was added to the reaction mixture to terminate the reaction, ethyl acetate and water were added, the organic layer was extracted, washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. Column chromatography (column: silica gel, mobile phase: normal) Purification with hexane / ethyl acetate = 1: 1) afforded the desired compound in 100% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.5-1.7 (m, 1H), 2.0-2.2 (m, 2H), 3.4-3.5 (m, 2H), 3.8-4.0 (m, 1H), 4.3-4.4 (q, J = 6.2, 14.7 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H), 7.2-7.4 (m, 2H), 7.7- 7.9 (dd, J = 12.6, 19.6 Hz, 2H). ; MS (EI, 70 eV) m / z 350 (M ⁺ )

Preparation Example 9: Allyl (2R, 4R) -2-{[4-amino- (1,3-benzothiazol-2-yl) sulfanyl] methyl} -4-hydroxy-1-pyrrolidine-1 -Carboxylic acid ester (compound of formula 9, X '= S, Y = 4-amino)

Allyl (2R, 4R) -2-{[(4-amino-1, 3-benzothiazol-2-yl) sulfanyl] methyl} -tert-butyldimethylsilyloxy-1-pyrrolidine-1-carboxylic acid The ester (compound of Preparation Example 3) (1.26 g, 2.63 mmol) was dissolved in 13 ml of tetrahydrofuran, acetic acid (1.51 ml, 26.3 mmol) and tetrahydroammonium fluoride (10.51 ml, 10.51 mmol / 1 mol tetrahydro) at 0 ° C. Furan solution) was added thereto, and then the temperature was raised to room temperature, followed by stirring for 18 hours. The reaction solution was treated in the same manner as in Preparation Example 8 to obtain the target compound in a yield of 86%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.2-1.3 (m, 1H), 2.0-2.2 (m, 2H), 3.4-3.5 (m, 2H), 3.6-3.8 (m, 1H), 4.3-4.4 (q, J = 7.2, 12.4 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H), 6.8-6.9 (dd, J = 12.6, 19.6 Hz, 1H), 6.9-7.0 (m, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H). ; MS (EI, 70 eV) m / z 365 (M ⁺ ), 196, 182, 170, 151, 137, 126, 108, 61.

Preparation Example 10: Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-hydroxy-pyrrolidinyl} methyl) sulfanyl] -1H-benzimidazole-1- Carboxylic Acid Ester (Compound of Formula 9, X '= N-allyloxycarbonyl, Y = H)

Allyl 2 - [({(2R, 4R) -1 - [( allyloxy) carbonyl] -4- tert-butyldimethylsilyloxy-pyrrolidinyl} methyl) sulfanyl] -1H- benzimidazole-1- The carboxylic acid ester (compound of Preparation Example 6) (1.50 g, 2.82 mmol) was dissolved in 15 ml of tetrahydrofuran, acetic acid (1.61 ml, 28.2 mmol) and tetrahydroammonium fluoride (11.281 ml, 11.28 mmol / 1 mol tetra) at 0 ° C. Hydrofuran solution) was added thereto, and then the temperature was raised to room temperature, followed by stirring for 18 hours. The reaction solution was treated as in Preparation Example 8 to obtain the target compound in a yield of 89%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.8-1.9 (m, 1H), 2.0-2.2 (m, 2H), 2.8-3.0 (m, 2H), 3.4-3.5 (dd, J = 6.4, 12.8 Hz , 2H), 3.5-3.6 (d, J = 6.2 Hz, 1H) 4.5-4.6 (m, 4H), 5.2-5.4 (m, 4H), 5.8-6.0 (m, 2H), 7.1-7.2 (m, 2H), 7.5-7.7 (dd, J = 1.2, 6.0 Hz, 1H), 7.8-7.9 (dd, J = 1.8, 7.6 Hz, 1H). ; MS (EI, 70 eV) m / z 417 (M ⁺ ), 360, 316, 298, 284, 259, 248, 234, 201, 190, 175, 150, 137, 126, 108, 90, 68.

Preparation Example 11: Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-hydroxy-pyrrolidinyl} methyl) sulfanyl] -4-nitro-1H-benzimi Dazole-1-carboxylic acid ester (compound of formula 9, X '= N-allyloxycarbonyl, Y = 4-nitro)

Allyl 2 - [({(2R, 4R) -1 - [( allyloxy) carbonyl] -4-tert-butyldimethylsilyloxy-pyrrolidinyl} methyl) sulfanyl] -4-nitro -1H- benzimidazole Dazole-1-carboxylic acid ester (compound of Preparation Example 7) (1.75 g, 3.04 mmol) was dissolved in 15 ml of tetrahydrofuran, acetic acid (1.61 ml, 28.2 mmol) and tetrahydroammonium fluoride (11.281 ml, 11.28 mmol) at 0 ° C. / 1 mol tetrahydrofuran solution) was added and the temperature was raised to room temperature and stirred for 18 hours. The reaction solution was treated as in Preparation Example 8 to obtain the target compound in 49% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.8-2.0 (m, 1H), 2.2-2.4 (m, 2H), 3.1 (s, 2H), 3.6-3.8 (m, 1H), 4.1-4.3 (m , 2H) 4.5-4.6 (m, 4H), 5.1-5.3 (q, J = 7.2, 12.4 Hz, 4H), 5.8-6.1 (m, 2H), 7.2-7.4 (dd, J = 6.8, 14.7 Hz, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H), 8.1-8.2 (dd, J = 6.6, 12.8 Hz, 1H). ; MS (EI, 70 eV) m / z 462 (M ⁺ ), 445, 405, 377, 361, 321, 267, 246, 220, 195, 183, 170, 154, 138, 126, 108, 80, 68.

Preparation Example 12: Allyl (2R, 4R) -4- (acetylsulfanyl) -2-[(1,3-benzothiazol-2-ylsulfanyl) methyl] -1-pyrrolidine-1-carboxylic acid ester (Compound of Formula 10, X '= S, Y = H)

Allyl (2R, 4R) -2-{[(1,3-benzothiazol-2-yl) sulfanyl] methyl} -4-hydroxy-1-pyrrolidine-1-carboxylic acid ester (of Preparation Example 8 Compound) (500 mg, 1.43 mmol) in 8.0 ml of tetrahydrofuran and triphenylphosphine (600 mg, 2.28 mmol) was added. Diethyl azodicarboxylate (0.38 ml, 2.42 mmol) was added at 0 ° C. and stirred for 30 minutes. At the same temperature, thioacetic acid (0.16 ml, 2.28 mmol) was added dropwise and stirred for 10 minutes, followed by stirring at room temperature for 20 minutes, and then the solvent was removed under reduced pressure. Purification by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 1) gave the desired compound in 42% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.5-1.7 (m, 1H), 2.0-2.2 (m, 2H), 2.3 (s, 3H), 3.4-3.5 (m, 2H), 3.8-4.0 (m , 1H), 4.3-4.4 (q, J = 6.2, 14.7 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H), 7.2-7.4 ( m, 2H), 7.7-7.9 (dd, J = 12.6, 19.6 Hz, 2H). ; MS (EI, 70 eV) m / z 408 (M ⁺ )

Preparation Example 13 Allyl (2R, 4R) -4- (acetylsulfanyl) -2-[(4-amino-1,3-benzothiazol-2-ylsulfanyl) methyl] -1-pyrrolidine- 1-carboxylic acid ester (compound of formula 10, X '= S, Y = 4-amino)

Allyl (2R, 4R) -2-{[4-amino- (1,3-benzothiazol-2-yl) sulfanyl] methyl} -4-hydroxy-1-pyrrolidine-1-carboxylic acid ester ( Compound 9) (822 mg, 2.25 mmol) in Preparation Example 9 was dissolved in 10 ml of tetrahydrofuran and triphenylphosphine (708 mg, 2.70 mmol) was added thereto. Diethyl azodicarboxylate (0.39 ml, 2.48 mmol) was added at 0 ° C. and stirred for 30 minutes. At the same temperature, thioacetic acid (0.21 ml, 2.93 mmol) was added dropwise and stirred for 10 minutes, followed by stirring at room temperature for 20 minutes, and then the solvent was removed under reduced pressure. Purification by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 1) gave the desired compound in a yield of 85%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.2-1.3 (m, 1H), 1.8-2.0 (m, 2H), 2.3 (s, 3H), 3.4-3.5 (m, 2H), 3.6-3.8 (m , 1H), 4.3-4.4 (q, J = 7.2, 12.4 Hz, 2H) 4.5-4.6 (m, 2H), 5.15-5.35 (m, 2H), 5.8-6.0 (m, 1H), 6.8-6.9 ( dd, J = 12.6, 19.6 Hz, 1H), 6.9-7.0 (m, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H). ; MS (EI, 70 eV) m / z 423 (M ⁺ ), 348, 242, 196, 182, 163, 152, 137, 123, 108, 80.

Preparation 14: Allyl 2-[({(2R, 4R) -4- (acetylsulfanyl) -1-[(allyloxy) carbonyl] pyrrolidinyl} methyl) sulfanyl] -1H-benzimidazole- 1-carboxylic acid ester (compound of formula 10, X '= N-allyloxycarbonyl, Y = H)

Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-hydroxy-pyrrolidinyl} methyl) sulfanyl] -1H-benzimidazole-1-carboxylic acid ester (manufacture Compound of Example 10) (1.05 g, 2.52 mmol) in 15 ml of tetrahydrofuran and triphenylphosphine (793 mg, 3.02 mmol) was added thereto. Diethyl azodicarboxylate (0.44 ml, 2.77 mmol) was added at 0 ° C. and stirred for 30 minutes. Thioacetic acid (0.23 ml, 3.28 mmol) was added dropwise at the same temperature, followed by stirring for 10 minutes, followed by stirring at room temperature for 20 minutes, and then the solvent was removed under reduced pressure. Purification by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 1) gave the desired compound in a yield of 78%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.8-1.9 (m, 1H), 2.0-2.2 (m, 2H), 2.3 (s, 3H), 2.8-3.0 (m, 2H), 3.4-3.5 (dd , J = 6.4, 12.8 Hz, 2H), 3.5-3.6 (d, J = 6.2 Hz, 1H) 4.5-4.6 (m, 4H), 5.2-5.4 (m, 4H), 5.8-6.0 (m, 2H) , 7.1-7.2 (m, 2H), 7.5-7.7 (dd, J = 1.2, 6.0 Hz, 1H), 7.8-7.9 (dd, J = 1.8, 7.6 Hz, 1H). ; MS (EI, 70 eV) m / z 475 (M ⁺ ), 432, 400, 356, 243, 234, 201, 190, 175, 166, 152, 131, 118, 108, 90, 80, 68.

Preparation 15: Allyl 2-[({(2R, 4R) -4- (acetylsulfanyl) -1-[(allyloxy) carbonyl] pyrrolidinyl} methyl) sulfanyl] -4-nitro-1H- Benzimidazole-1-carboxylic acid ester (compound of formula 10, X '= N-allyloxycarbonyl, Y = 4-nitro)

Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-hydroxy-pyrrolidinyl} methyl) sulfanyl] -4-nitro-1H-benzimidazole-1- The carboxylic acid ester (compound of Preparation Example 11) (656 mg, 1.42 mmol) was dissolved in 15 ml of tetrahydrofuran and triphenylphosphine (793 mg, 3.02 mmol) was added thereto. Diethyl azodicarboxylate (0.44 ml, 2.77 mmol) was added at 0 ° C. and stirred for 30 minutes. Thioacetic acid (0.23 ml, 3.28 mmol) was added dropwise at the same temperature, followed by stirring for 10 minutes, followed by stirring at room temperature for 20 minutes, and then the solvent was removed under reduced pressure. Purification by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 1) gave the desired compound in a yield of 59%.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.8-2.0 (m, 1H), 2.3 (s, 3H), 2.3-2.4 (m, 2H), 3.1 (s, 2H), 3.6-3.8 (m, 1H ), 4.1-4.3 (m, 2H) 4.5-4.6 (m, 4H), 5.1-5.3 (q, J = 7.2, 12.4 Hz, 4H), 5.8-6.1 (m, 2H), 7.2-7.4 (dd, J = 6.8, 14.7 Hz, 1H), 7.5-7.7 (d, J = 6.4 Hz, 1H), 8.1-8.2 (dd, J = 6.6, 12.8 Hz, 1H). ; MS (EI, 70 eV) m / z 520 (M ⁺ ), 432, 377, 361, 243, 201, 190, 166, 153, 131, 118, 108, 80, 68.

Preparation Example 16 allyl (2R, 4R) -2-[(1,3-benzothiazol-2-ylsulfanyl) methyl] -4-mergato-1-pyrrolidinecarboxylic acid ester (compound of formula 3, X '= S, Y = H)

Allyl (2R, 4R) -4- (acetylsulfanyl) -2-[(1,3-benzothiazol-2-ylsulfanyl) methyl] -1-pyrrolidine-1-carboxylic acid ester (Preparation Example 12 Compound) (250 mg, 0.59 mmol) was dissolved in 1.3 ml of methanol, 2N-sodium hydroxide (0.30 ml, 0.59 mmol) was added at 0 ° C., and stirred for 20 minutes. To the reaction solution was added a mixed solution of acetic acid (35 mg, 0.59 mmol), ethyl acetate (15 ml) and water (5 ml) to terminate the reaction. The organic layer was extracted and washed with 10% aqueous sodium bicarbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure and used directly in the next reaction.

TLC: R f = 0.80 (ethyl acetate / n-hexane = 3/1)

Preparation Example 17: Allyl (2R, 4R) -2-[(4-amino-1, 3-benzothiazol-2-ylsulfanyl) methyl] -4-mercapto-1-pyrrolidinecarboxylic acid ester 3 compound, X '= S, Y = 4-amino)

Allyl (2R, 4R) -4- (acetylsulfanyl) -2-[(4-amino-1,3-benzothiazol-2-ylsulfanyl) methyl] -1-pyrrolidine-1-carboxylic acid ester (Compound of Preparation Example 13) (814 mg, 1.92 mmol) was dissolved in 8 ml of methanol, 2N-sodium hydroxide (1.06 ml, 2.11 mmol) was added at 0 ° C., and stirred for 20 minutes. Acetic acid (125 mg, 2.11 mmol), a mixed solution of ethyl acetate (30 ml) and water (10 ml) were added to the reaction mixture to complete the reaction. The organic layer was extracted and washed with 10% aqueous sodium bicarbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure and used directly in the next reaction.

TLC: R f = 0.79 (ethyl acetate / n-hexane = 3/1)

Preparation Example 18: Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-mercaptopyrrolidinyl} methyl) sulfanyl] -1H-benzimidazole-1-carboxylic acid Esters (compound of formula 3, X '= N-allyloxycarbonyl, Y = H)

Allyl 2-[({(2R, 4R) -4- (acetylsulfanyl) -1-[(allyloxy) carbonyl] pyrrolidinyl} methyl) sulfanyl] -1H-benzimidazole-1-carboxylic acid ester (Compound of Production Example 14) (555 mg, 1.17 mmol) was dissolved in 5 ml of methanol, 2N-sodium hydroxide (0.52 ml, 0.59 mmol) was added at 0 ° C., and stirred for 20 minutes. Acetic acid (33 mg, 0.59 mmol), ethyl acetate (15 ml) and water (5 ml) were added to the reaction mixture to terminate the reaction. The organic layer was extracted and washed with 10% aqueous sodium bicarbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure and used directly in the next reaction.

TLC: R f = 0.40 (ethyl acetate / n-hexane = 1/1)

Preparation Example 19: Allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-mercaptopyrrolidinyl} methyl) sulfanyl] -4-nitro-1H-benzimidazole -1-carboxylic acid ester (compound of formula 3, X ′ = N-allyloxycarbonyl, Y = 4-nitro)

2-[({(2R, 4R) -4- (acetylsulfanyl) -1-[(allyloxy) carbonyl] pyrrolidinyl} methyl) sulfanyl] -4-nitro-1H-benzimidazole-1 -Carboxylic acid ester (compound of Preparation 15) (200 mg, 0.38 mmol) was dissolved in 2 ml of methanol, 2N-sodium hydroxide (0.16 ml, 0.32 mmol) was added at 0 ° C, and stirred for 20 minutes. A mixed solution of acetic acid (0.32 mmol), ethyl acetate (10 ml) and water (5 ml) was added to the reaction mixture to complete the reaction. The organic layer was extracted, and washed with 10% aqueous sodium bicarbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure and used directly in the next reaction.

TLC: R f = 0.44 (ethyl acetate / n-hexane = 1/1)

Example 1: Allyl ( 4S , 6R ) -3-({( 3S , 5R ) -1-[(allyloxy) carbonyl] -5-[(1,3-benzthiazole-2- ylsulfanyl) methyl] pyrrolidinyl} sulfanyl) -6 - [(1 S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo cyclo [3.2.0] hept-2 En-2-carboxylic acid ester (compound of formula 4, R ₁ = H, R ₂ = allyl, R ₃ = allyloxycarbonyl, X '= S, Y = H)

Allyl (1R, 5R, 6S) -6-[(1R) -1-tert-butyldimethylsilyloxyethyl] -1-methyl-2-diphenylphosphoryloxy-2-carbafen-2-m-3- The carboxylic ester (compound of formula 2) (299 mg, 0.60 mmol) was dissolved in 1.5 mL of acetonitrile and allyl (2R, 4R) -2-[(1,3-benzothiazol-2-ylsulfanyl) methyl at 0 ° C. ] -4-mercapto-1-pyrrolidinecarboxylic acid ester (compound of Preparation 16) (200 mg, 0.55 mmol) was added dropwise to 1.5 mL of acetonitrile, and then diisopropylethylamine (0.19 ml, 1.09 mmol) was added. ) Was added. The mixture was stirred at the same temperature for 20 minutes and then stirred at room temperature for 2 hours to complete the reaction. An appropriate amount of ethyl acetate and water were added to the reaction mixture, the organic layer was extracted, washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 2: 1) to afford the desired compound in 35% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.16-1.18 (d, J = 7.2 Hz, 3H), 1.27-1.29 (d, J = 6.2 Hz, 3H), 1.79-1.97 (m, 2H), 3.17 ( m, 1H), 3.27 (m, 1H), 3.27-3.28 (t, J = 2.9 Hz, 2H), 4.0-4.1 (m, 2H), 4.14-4.17 (d, J = 6.4 Hz, 2H) 4.56- 4.57 (m, 1H), 4.58-4.64 (t, J = 5.6, 19.0 Hz, 1H) 4.65-4.67 (m, 4H), 5.13-5.33 (m, 4H), 5.84-5.88 (m, 2H), 7.23 -7.34 (m, 2 H), 7.67-7.70 (m, 2 H). ; MS (EI, 70 eV) m / z 615 (M ⁺ )

Example 2 : Allyl ( 4S , 6R ) -3-({( 3S , 5R ) -1-[(allyloxy) carbonyl] -5-[(4-amino-1,3-benzothia -2-ylsulfanyl) methyl] pyrrolidinyl} sulfanyl) -6 - [(1 S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo cyclo [3.2.0] Hept-2-ene-2-carboxylic acid ester (compound of formula 4, R ₁ = H, R ₂ = allyl, R ₃ = allyloxycarbonyl, X '= S, Y = 4-amino)

Allyl (1R, 5R, 6S) -6 - [(1R) -1- tert - butyldimethylsilyloxy ethyl] -1-methyl-2-diphenylphosphoryl-2-pen-2-em-3-carbazole The carboxylic acid ester (compound of formula 2) (872 mg, 1.75 mmol) was dissolved in 10 mL of acetonitrile and allyl (2R, 4R) -2-[(4-nitro-1,3-benzothiazol-2-ylsulfa at 0 ° C. Nil) methyl] -4-mercapto-1-pyrrolidinecarboxylic acid ester (compound of Preparation 17) (1.92 mmol) was added dropwise to 5 mL of acetonitrile, and then diisopropylethylamine (0.61 ml. 3.50 Mmol). After stirring for 20 minutes at the same temperature, the mixture was stirred for 2 hours at room temperature to complete the reaction. An appropriate amount of ethyl acetate and water were added to the reaction mixture, the organic layer was extracted, washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 2) to afford the desired compound in 13% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.16-1.17 (d, J = 7.0 Hz, 3H), 1.27-1.29 (d, J = 6.2 Hz, 3H), 1.82-1.91 (m, 2H), 3.1 ( m, 1H), 3.21 (m, 1H), 3.27-3.28 (t, J = 4.2 Hz, 2H), 4.0-4.1 (m, 2H), 4.14-4.17 (d, J = 6.4 Hz, 2H) 4.56- 4.57 (m, 1H), 4.58-4.64 (t, J = 5.6, 19.0 Hz, 1H) 4.65-4.67 (m, 4H), 5.13-5.33 (m, 4H), 5.88-6.00 (m, 2H), 7.23 -7.26 (m, 2 H), 7.67-7.70 (dd, J = 8.6 Hz, 1 H). ; MS (EI, 70 eV) m / z 630 (M ⁺ ).

Example 3: Allyl 2-({[( 2R , 4S ) -1-[(allyloxy) carbonyl] -4-({( 4S , 6R ) -2-[(allyloxy) carbonyl ] -6-[( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-3-yl} sulfanyl) pyrrolidinyl ] Methyl} sulfanyl) -1 H -benzimidazole-1-carboxylic acid ester (compound of formula 4, R ₁ = H, R ₂ = allyl, R ₃ = allyloxycarbonyl, X '= N-allyloxycarbono Neal, Y = H)

Allyl (1R, 5R, 6S) -6 - [(1R) -1- tert - butyldimethylsilyloxy ethyl] -1-methyl-2-diphenylphosphoryl-2-pen-2-em-3-carbazole The carboxylic acid ester (compound of formula 2) (531 mg, 1.06 mmol) was dissolved in 6 mL of acetonitrile and allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-mergatopi at 0 ° C. A solution obtained by dissolving ralidinyl} methyl) sulfanyl] -1H-benzimidazole-1-carboxylic acid ester (compound of Preparation 18) (1.17 mmol) in 5 ml of acetonitrile was added, followed by diisopropylethylamine (0.61 ml. 3.50 Mmol). After stirring for 20 minutes at the same temperature, the mixture was stirred for 2 hours at room temperature to complete the reaction. An appropriate amount of ethyl acetate and water were added to the reaction mixture, the organic layer was extracted, washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 2) to afford the desired compound in 30% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.15-1.19 (td, J = 6.0, 6.8 Hz, 3H), 1.33-1.36 (d, J = 6.0 Hz, 3H), 1.8-2.0 (m, 2H), 3.1-3.2 (t, J = 7.1 Hz, 3H), 3.7-3.9 (m, 3H), 4.0-4.1 (t, J = 6.4 Hz, 2H), 4.14-4.17 (m, 2H) 4.6-4.7 (m , 3H), 4.75-4.95 (dd, J = 6.4 Hz, 3H) 5.09-5.2 (t, J = 7.2 Hz, 3H), 5.2-5.4 (d, J = 6.0 Hz, 3H), 5.8-6.0 (m , 3H), 7.15-7.20 (m, 2H), 7.40-7.60 (m, 2H). ; MS (EI, 70 eV) m / z 682 (M ⁺ ), 554, 513, 405, 363, 349, 316, 271, 240, 200, 186, 166, 150, 119, 82, 68.

Example 4: Allyl 2-({[( 2R , 4S ) -1-[(allyloxy) carbonyl] -4-({( 4S , 6R ) -2-[(allyloxy) carbonyl ] -6-[( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-3-yl} sulfanyl) pyrrolidinyl ] Methyl} sulfanyl) -4-nitro-1 H -benzimidazole-1-carboxylic acid ester ( compound of formula 4, R ₁ = H, R ₂ = allyl, R ₃ = allyloxycarbonyl, X '= N Allyloxycarbonyl, Y = 4-nitro)

Allyl (1R, 5R, 6S) -6 - [(1R) -1- tert - butyldimethylsilyloxy ethyl] -1-methyl-2-diphenylphosphoryl-2-pen-2-em-3-carbazole The carboxylic ester (compound of formula 2) (173 mg, 0.35 mmol) was dissolved in 2 mL of acetonitrile and allyl 2-[({(2R, 4R) -1-[(allyloxy) carbonyl] -4-mer at 0 ° C. Lecaptopyrrolidinyl} methyl) sulfanyl] -4-nitro-1H-benzimidazole-1-carboxylic acid ester (compound of Preparation 19) (0.38 mmol) in 1 ml of acetonitrile was added, followed by diisopropylethyl Amine (0.12 ml. 0.7 mmol) was added. After stirring for 20 minutes at the same temperature, the mixture was stirred for 2 hours at room temperature to complete the reaction. An appropriate amount of ethyl acetate and water were added to the reaction mixture, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (column: silica gel, mobile phase: normal hexane / ethyl acetate = 1: 2) to afford the desired compound in 47% yield.

¹ H NMR (200 MHz, CDCl ₃ ) δ 1.2-1.25 (d, J = 6.4 Hz, 3H), 1.30-1.35 (d, J = 6.4 Hz, 3H), 1.80-1.85 (m, 2H), 3.10- 3.15 (t, J = 6.8 Hz, 3H), 3.7-3.9 (m, 3H), 4.0-4.1 (t, J = 6.4 Hz, 2H), 4.12-4.15 (m, 2H) 4.4-4.5 (m, 3H ), 4.70-4.85 (dd, J = 6.4 Hz, 3H) 5.0-5.2 (t, J = 7.4 Hz, 3H), 5.3-5.4 (d, J = 6.2 Hz, 3H), 5.8-6.0 (m, 3H ), 7.20-7.23 (m, 2H), 7.4-7.6 (m, 1H). ; MS (EI, 70 eV) m / z 727 (M ⁺ ) 554, 513, 405, 363, 349, 316, 271, 240, 200, 186, 166, 150, 119, 82, 68.

Example 5: Preparation of sodium (4 S, 6 R) -3 - ({(3 S, 5 R) -5 - [(1,3- benzothiazol-2-ylsulfanyl) methyl] pyrrolidinyl} sulfanyl Nil) -6-[( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (compound of formula 1) , X = S, Y = H)

Allyl ( 4S , 6R ) -3-({( 3S , 5R ) -1-[(allyloxy) carbonyl] -5-[(1,3-benzthiazol-2-ylsulfanyl) methyl] pyrrolidinyl} sulfanyl) -6 - [(1 S) -1- hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo cyclo [3.2.0] hept-2-ene-2- The carboxylic ester (compound of Example 1) (97 mg, 0.16 mmol) was dissolved in 1 mL of dichloromethane and triphenylphosphine (8.26 mg, 0.032 mmol), tetrakis (triphenylphosphine) palladium (0) (9.61 mg, 0.008 Mmol) and sodium 2-ethylhexanoate (53.18 mg, 0.32 mmol) were added sequentially and stirred at room temperature for 4 hours. 3 ml of water was added to the reaction solution, the organic solvent was removed under reduced pressure, and the remaining aqueous solution was washed with diethyl ester, followed by removing an insoluble solid. The aqueous solution obtained by purification by C18 reversed phase column chromatography (mobile phase: 10% acetonitrile / water) was lyophilized to give a pale off-white target compound in a yield of 60%.

¹ H NMR (300 MHz, D ₂ O) δ 1.13-1.15 (d, J = 7.0 Hz, 3H), 1.2-1.21 (d, J = 6.4 Hz, 3H), 1.65-1.77 (m, 2H), 3.15 (m, 1H), 3.25 (m, 1H), 3.27-3.28 (t, J = 2.9 Hz, 2H), 4.0-4.1 (m, 2H), 4.12-4.14 (d, J = 6.4 Hz, 2H) 4.56 -4.57 (m, 1H), 4.59-4.63 (t, J = 6.0, 12.0 Hz, 1H) 7.2-7.3 (m, 2H), 7.6-7.7 (m, 2H). ; MS (EI, 70 eV) m / z 535 (M ⁺ ).

Example 6: Sodium ( 4S , 6R ) -3-({( 3S , 5R ) -5-[(4-amino-1,3-benzothiazol-2-ylsulfanyl) methyl] pi Lolidinyl} sulfanyl) -6-[( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate ( Compound of Formula 1, X = S, Y = 4-nitro)

Allyl ( 4S , 6R ) -3-({( 3S , 5R ) -1-[(allyloxy) carbonyl] -5-[(4-nitro-1,3-benzthiazol-2- ylsulfanyl) methyl] pyrrolidinyl} sulfanyl) -6 - [(1 S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo cyclo [3.2.0] hept-2 En-2-carboxylic acid ester (compound of Example 2) (149 mg, 0.24 mmol) was dissolved in 2 mL of dichloromethane, triphenylphosphine (12.4 mg, 0.047 mmol), tetrakis (triphenylphosphine) palladium (0) ( 14.4 mg, 0.012 mmol) and sodium 2-ethylhexanoate (79.8 mg, 0.48 mmol) were added in this order and stirred at room temperature for 4 hours. 6 ml of water was added to the reaction solution, the organic solvent was removed under reduced pressure, and the remaining aqueous solution was washed with diethyl ester to remove an insoluble solid. The aqueous solution obtained by purification by C18 reversed phase column chromatography (mobile phase: 10% acetonitrile / water) was lyophilized to give a pale off-white target compound in 25% yield.

¹ H NMR (300 MHz, D ₂ O) δ 1.15-1.16 (d, J = 7.2 Hz, 3H), 1.26-1.29 (d, J = 6.4 Hz, 3H), 1.8-1.9 (m, 2H), 3.20 -3.23 (m, 1H), 3.23-3.24 (m, 1H), 3.28-3.30 (t, J = 4.6 Hz, 2H), 4.0-4.1 (m, 2H), 4.20-4.22 (d, J = 6.4 Hz , 2H) 4.5-4.6 (m, 1H), 4.58-4.62 (t, J = 6.2, 14.8 Hz, 1H) 7.23-7.26 (m, 2H), 7.60-7.65 (dd, J = 7.6 Hz, 1H). ; MS (EI, 70 eV) m / z 550 (M ⁺ ).

Example 7: Sodium ( 4S , 6R ) -3-({( 3S , 5R ) -5-[( 1H -benzimidazol-2-ylsulfanyl) methyl] pyrrolidinyl} sulfanyl ) -6-[( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (compound of formula 1, X = N, Y = H)

Allyl 2-({[( 2R , 4S ) -1-[(allyloxy) carbonyl] -4-({( 4S , 6R ) -2-[(allyloxy) carbonyl] -6- [( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-3-yl} sulfanyl) pyrrolidinyl] methyl} sulpa carbonyl) -1 H - benzimidazole-1-carboxylic acid ester of the compound (example 3) (212 mg, 0.31 mmol) was dissolved in 2 mL of dichloromethane and triphenylphosphine (24.39 mg, 0.093 mmol), tetrakis (triphenylphosphine) palladium (0) (27.94mg, 0.024 mmol), sodium 2-ethylhexano Eight (155 mg, 0.93 mmol) was added sequentially, and it stirred at room temperature for 4 hours. 6 ml of water was added to the reaction solution, the organic solvent was removed under reduced pressure, and the remaining aqueous solution was washed with diethyl ester to remove an insoluble solid. The aqueous solution obtained by purification by C18 reversed phase column chromatography (mobile phase: 10% acetonitrile / water) was lyophilized to give a pale off-white target compound in a yield of 39%.

¹ H NMR (300 MHz, D ₂ O) δ 1.16-1.18 (td, J = 6.2, 6.4 Hz, 3H), 1.3-1.32 (d, J = 6.4 Hz, 3H), 1.8-2.1 (m, 2H) , 3.15-3.22 (t, J = 7.4 Hz, 3H), 3.75-3.90 (m, 3H), 4.2-4.25 (t, J = 6.4 Hz, 2H), 4.3-4.4 (m, 2H) 7.1-7.2 ( m, 2H), 7.3-7.5 (m, 2H). ; MS (EI, 70 eV) m / z 640 (M ⁺ ).

Example 8: Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1H-4-nitrobenzimidazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl ) -6-[(1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (Compound 1, X = N, Y = 4-nitro)

Allyl 2-({[( 2R , 4S ) -1-[(allyloxy) carbonyl] -4-({( 4S , 6R ) -2-[(allyloxy) carbonyl] -6- [( 1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-3-yl} sulfanyl) pyrrolidinyl] methyl} sulpa Nil) -4-nitro-1 H -benzimidazole-1-carboxylic acid ester (compound of Example 4) (153.4 mg, 0.21 mmol) was dissolved in 2 mL of dichloromethane and triphenylphosphine (16.59 mg, 0.063 mmol), tetrakis (triphenylphosphine) palladium (0) (18.93mg, 0.016 mmol), sodium 2-ethylhexa Noate (105 mg, 0.63 mmol) was added sequentially and stirred at room temperature for 4 hours. 6 ml of water was added to the reaction solution, the organic solvent was removed under reduced pressure, and the remaining aqueous solution was washed with diethyl ester to remove an insoluble solid. The aqueous solution obtained by purification by C18 reverse phase column chromatography (mobile phase: 10% acetonitrile / water) was lyophilized to give a pale off-white target compound in a yield of 29%.

¹ H NMR (300 MHz, D ₂ O) δ 1.18-1.20 (d, J = 6.4 Hz, 3H), 1.3-1.4 (d, J = 6.4 Hz, 3H), 1.8-1.9 (m, 2H), 3.0 -3.15 (t, J = 7.2 Hz, 3H), 3.7-3.8 (m, 3H), 4.1-4.2 (t, J = 6.4 Hz, 2H), 4.20-4.25 (m, 2H) 7.2-7.3 (m, 2H), 7.4-7.5 (m, 1 H). ; MS (EI, 70 eV) m / z 685 (M ⁺ ).

Test Example 1

The antimicrobial activity of Gram-positive bacteria, Methicillin-resistant strains (MRSA) and Ofloxacin-resistant strains (QRSA), of the compound represented by Formula 1 according to the present invention was tested as follows.

The present invention prepared in Examples 5 to 8 by measuring the Minimum Inhibitory Concentration (MIC) according to the agar medium dilution method by double agar dilution method using Muller-Hinton agar. The antimicrobial activity of the carbapenem derivatives of the present invention was evaluated and the results are shown in Table 1 below. As a control group, meropenem (Yuhan Corporation) and vancomycin (Cheil Jedang) were used for MRSA infection.

As can be seen from Table 1, the carbapenem derivative of the formula (1) of the present invention shows an excellent antimicrobial activity against MRSA and QRSA compared to the control, especially the compound of Example 5 showed an excellent antimicrobial activity.

Test Example 2

In order to measure the in vivo antimicrobial and pharmacological activity of the carbapenem derivatives of the present invention, Streptococcus Pyogenes 77A was used as a base strain, and it was 4 to 5 weeks old as a test animal. ICR-based mice with a body weight of 22 to 25 g were used. The compound prepared in Example 5 was dissolved in distilled water and administered to the mice by SC so that the amount of the test compound was 40 mg / kg (mouse). As a control, meropenem (Yuhan Corporation), a commercial carbapenem antibiotic, was used. Blood was collected directly from the heart of mice before, 1, 2, 4, 6, 8, and 24 hours after administration, respectively.

Plasma was obtained by centrifuging blood at 12,000 rpm, and 400 µl of acetonitrile internal standard was added to 200 µl of plasma, followed by shaking extraction. The extract was centrifuged at 1,000 rpm, and 50 ml of the supernatant was taken and measured by micro high pressure liquid chromatography (HPLC).

The change over time (C _max , T _1/2 , and AUC) of the administered drug concentration in mouse blood was measured and the results are shown in Table 2 below.

division AUC ^{* 1} (μg.h / ml) T _max ^{* 2} (hr) C _max ^{* 3} (µg / ml) T _1/2 ^{* 4} (hr) Example 5 13.04 0.17 0.48 8.02 Meropenem 14.07 0.25 0.28 7.82 * 1: Area under the blood concentration curve up to 24 hours after dosing * 2: Time at maximum blood concentration * 3: Maximum blood concentration * 4: Time at 1/2 blood concentration

As can be seen from Table 2, the compound of Example 5, the carbapenem derivative of the present invention exhibits the same bioavailability as that of the control meropenem, it is easy to use as a drug.

The carbapenem derivative of the present invention has excellent antibacterial effect against methicillin resistant strain (MRSA) and oploxacin resistant strain (QRSA), and can be usefully used as an antibiotic against intractable resistant bacterial infections including MRSA and QRSA.

Claims (5)

Carbapenem derivative of Formula 1 or a pharmacologically acceptable salt thereof: Formula 1 Where X represents a sulfur or nitrogen atom; Y represents a hydrogen, nitro group or amine group present at a specific position of the phenyl group; M represents a hydrogen ion or a counterion forming a pharmacologically acceptable salt. The method of claim 1, Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1,3-benzothiazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6- [ (1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, Sodium (4S, 6R) -3-({(3S, 5R) -5-[(4-amino-1,3-benzothiazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6-[(1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1H-benzimidazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6-[(1S ) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, or Sodium (4S, 6R) -3-({(3S, 5R) -5-[(1H-4-nitrobenzimidazol-2-yl-sulfanyl) methyl] -pyrrolidinyl} sulfanyl) -6- A derivative that is [(1S) -1-hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate or a pharmacologically acceptable salt thereof. Reacting a compound of Formula 2 with a compound of Formula 3 to prepare a carbapenem ester derivative of Formula 4, and then removing the carboxy protecting group, amine protecting group and hydroxy protecting group from the compound of Formula 4, Method for preparing a carbapenem derivative of claim: Formula 2 Formula 3 Formula 4 In which R ₁ is a hydrogen atom or a hydroxy protecting group; R ₂ is a carboxy protecting group; R ₃ is an amine protecting group; X 'is a sulfur atom or a nitrogen atom protected with an amine protecting group; Y is a hydrogen, nitro group or amine group present at a specific position of the phenyl group. The method of claim 3, wherein Compound of formula (3), i) preparing a methyl iodide compound of formula 6 by refluxing the compound of formula 5 with sodium iodide in an acetone solvent for 40 to 48 hours; ii) preparing a compound of formula 8 by heating and refluxing the compound of formula 6 with potassium carbonate and a thiol compound of formula 7 in an organic solvent for 3 to 5 hours, wherein X in the compound of formula 7 is a nitrogen atom In the methylene chloride solvent, the resulting compound is reacted with triethylamine and allyloxycarbonyl chloride, p-nitrobenzyloxycarbonyl chloride or p-methoxybenzyloxycarbonyl chloride at 0 ° C. for 2 hours. Preparing a compound of formula 8 protected with an amine protecting group, iii) reacting the compound of formula 8 with tetrabutylammonium fluoride in a tetrahydrofuran solvent at room temperature for 18 hours to prepare a compound of formula 9, iv) The compound of formula 9 is reacted with triphenylphosphine and diethylazodicarboxylic acid or diisopropylazocarboxylic acid in an organic solvent at -5 to 5 DEG C for 30 minutes to 2 hours, and then thioacetic acid is added thereto at room temperature. Reacting for 1 to 3 hours to prepare a compound of Formula 10, v) a process prepared by a process comprising the step of reacting a compound of formula 10 with a base to hydrolyze: Where X is a sulfur or nitrogen atom, R ₃ , X 'and Y are as defined in claim 3. An antibiotic composition comprising an effective amount of a carbapenem derivative of claim 1 or a pharmacologically acceptable salt thereof, and a pharmaceutically acceptable carrier.