KR930004012B1 - Process for preparing penem derivatives - Google Patents

Abstract

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Description

Method for preparing penem derivative

The present invention relates to a novel penem derivative of the general formula (I) and a pharmaceutically acceptable salt thereof having excellent antibiotic activity.

또는

의 치환되거나 비치환된 비사이클로 헤테로사이클릭 그룹이고;Wherein R ₁ is hydrogen, C ₁ to C ₆ alkyl or hydroxyl C ₁ to C ₆ alkyl group; COOR ₂ is a carboxyl group or carboxylate anion; R ₂ may be an ester moiety which may simultaneously act as a protecting group; R ₃ is a general formula

or

A substituted or unsubstituted bicyclo heterocyclic group of;

이며; 일반식

의 R₃의 부분구조는 5- 또는 6-원 질소-함유 헤테로사이클환 또는 4급 질소-함유 헤테로사이클환이고, 이때 질소-함유 헤테로사이클환은 산소, 황 및 질소 중에서 선택된 1 내지 4개의 헤테로원자를 갖는 포화 또는 불포화된 헤테로사이클환이며; R₄는 할로겐, 아미노, 하이드록실, 시아노, 임의로 치환된 C₁내지 C₆알콕실, 임의로 치환된 카바모일, 임의로 치환된 C₁내지 C₆알킬, 임의로 치환된 C₂내지 C₆알케닐, 임의로 치한된 C₂내지 C₆알키닐, 임의로 치환된 C₃내지 C₆사이클로알킬, 임의로 치환된 C₃내지 C₆사이클로알케닐, 임의로 치환된 C₃내지 C₆사이클로알킬-C₁, 내지 C₆알킬, 임의로 치환된 C₃내지 C₆사이클로알케닐-C₁내지 C₆알킬, 임의로 치환된 C₃내지 C₆사이클로알케닐-C₂내지 C₆알케닐, 임의로 치환된 헤테로사이클릴(헤테로사이클릴이란 산소, 질소 및 황 중에서 선택된 1 내지 4개의 헤테로원자 및 1 내지 6개의 탄소원자를 함유하는 3원 내지 7원 사이클릭그룹을 의미한다), 임의로 치환된 헤테로 사이클릴-C₁내지 C₆알킬, 임의로 치환된 헤테로사이클릴-C₂내지 C₆알케닐, 임의로 치환된 헤테로사이클릴-C₂내지 C₆알키닐, 임의로 치환된 C₃내지 C₆사이클로알킬리덴-C₁내지 C₆알킬, 임의로 치환된 C₃내지 C₆헤테로사이클릴리덴-C₁내지 C₆알킬, 임의로 치환된 아릴, 임의로 치환된 아릴-C₁내지 C₆알킬, 임의로 치환된 아릴-C₂내지 C₆알케닐, 임의로 치한된 아릴- C₂내지 C₆알키닐, 또는 일반식 R₄₄-(CH₂)v-Z-(CH₂)w-의 그룹이고;Where m is 1,2 or 3; Q is

Is; General formula

The substructure of R ₃ is a 5- or 6-membered nitrogen-containing heterocycle ring or a quaternary nitrogen-containing heterocycle ring, wherein the nitrogen-containing heterocycle ring is 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen. Saturated or unsaturated heterocycle ring having; R ₄ is halogen, amino, hydroxyl, cyano, optionally substituted C ₁ to C ₆ alkoxyl, optionally substituted carbamoyl, optionally substituted C ₁ to C ₆ alkyl, optionally substituted C ₂ to C ₆ alkenyl , Optionally substituted C ₂ to C ₆ alkynyl, optionally substituted C ₃ to C ₆ cycloalkyl, optionally substituted C ₃ to C ₆ cycloalkenyl, optionally substituted C ₃ to C ₆ cycloalkyl-C ₁ , to C ₆ alkyl, optionally substituted C ₃ to C ₆ cycloalkenyl-C ₁ to C ₆ alkyl, optionally substituted C ₃ to C ₆ cycloalkenyl-C ₂ to C ₆ alkenyl, optionally substituted heterocyclyl ( Heterocyclyl means a 3 to 7 membered cyclic group containing 1 to 4 heteroatoms and 1 to 6 carbon atoms selected from oxygen, nitrogen and sulfur), optionally substituted heterocyclyl-C ₁ to C ₆ alkyl, optionally substituted heterocyclyl-C ₂ to C ₆ Alkenyl, optionally substituted heterocyclyl-C ₂ to C ₆ alkynyl, optionally substituted C ₃ to C ₆ cycloalkylidene-C ₁ to C ₆ alkyl, optionally substituted C ₃ to C ₆ heterocyclylidene- C ₁ to C ₆ alkyl, optionally substituted aryl, optionally substituted aryl-C ₁ to C ₆ alkyl, optionally substituted aryl-C ₂ to C ₆ alkenyl, optionally substituted aryl- C ₂ to C ₆ alkynyl, Or a group of formula R _44- (CH ₂ ) vZ— (CH ₂ ) w—;

(Rb)(Rc)(여기서, Ra,Rb 및 Rc는 상기에서 정의한 바와 같다)의 카밤이미도일그룹, 일반식

(여기서, Ra,Rb,Rc 및 Rd는 각각 수소 또는 임의로 치환된 C₁내지 C₆알킬이거나, 이들중 2개가 함께는 C₃내지 C₆사이클로알킬 또는 헤테로사이클릴을 형성할 수 있다)의 구아니디닐그룹, 임의로 치환된 페닐, 임의로 치환된 헤테로사이클릴, C₁내지 C₆알킬 및 임의로 치환된 C₁내지 C₆알킬(여기서, 치환기는 아미노, 카복실, 시아노, 설포, 할로게노, 카보닐, 트리플루오로메틸, 니트로, 하이드록실, 포르밀, 모노-(임의로 치환된) C₁내지 C₆알킬아미노, 디-(임의로 치환된) C₁내지 C₆알킬아미노 3급-(임의로 치환된) C₁내지 C₆알킬암모니오, 카바모일, 모노- 또는 디-C₁내지 C₆알킬 카바모일, 카바모일옥시, 모노- 또는 디-C₁내지 C₆알킬 카바모일옥시, 일반식 -N(Ra)-C(Rb)=NRc(여기서, Ra,Rb 및 Rc는 각각 수소 또는 임의로 치환된 C₁내지 C₆알킬이거나, 이들중 인접한 2개가 함께는 C₃내지 C₆사이클로알킬 또는 헤테로사이클릴을 형성할 수 있다)의 아미디노그룹, 및 일반식

(여기서, Ra,Rb 및 Rc는 상기에서 정의한 바와 같다)의 카밤이미도일 그룹.Wherein Z is oxygen, sulfur SO, SO ₂ or NH; v is 0, 1 or 2; w is 0, 1, 2 or 3; R ₄₄ is hydrogen or C ₁ to C ₆ alkyl, C ₂ to C ₆ alkenyl, C ₂ to C ₆ alkynyl, C ₃ to C ₆ cycloalkyl, C ₃ to C ₆ cycloalkenyl, aryl, heterocycle A moiety selected from aryl, heterocyclyl-C ₁ to C ₆ alkyl, heterocyclyl-C ₂ to C ₆ alkenyl, and heterocyclyl-C ₂ to C ₆ alkynyl, each of which is optionally substituted; Optional substituents on R ₄ are amino, mono- (optionally substituted) C ₁ to C ₆ alkylamino, di- (optionally substituted) C ₁ to C ₆ alkylamino, tri- (optionally substituted) C ₁ to C ₆ alkylammonio, halogeno, carbonyl, trifluoromethyl, nitro, hydroxyl, formyl, cyano, formylamino, carboxyl, azido, sulfo, imino, carbamoyl, formamido, mono- Or di-C ₁ to C ₆ alkylcarbamoyl, formamide, mono- or di-C ₁ to C ₆ alkylcarbamoyl, carbamoyloxy, mono- or di-C ₁ to C ₆ alkylcarbamoyloxy, optionally substituted C ₁ to C ₆ alkoxy, optionally substituted C ₁ to C ₆ alkoxycarbonyl, optionally substituted C ₁ to C ₆ alkylcarbonyloxy, optionally substituted C ₁ to C ₆ alkylthio, sulfinyl, mono- or Di-C ₁ to C ₆ alkylsulfinyl, sulfonyl, mono or di-C ₁ to C ₆ alkylsulfinyl, sulfonyl, mono or di-C ₁ to C ₆ Alkylsulfonyl, sulfamoyl, mono- or di-C ₁ to C ₆ alkylsulfamoyl, sulfinamoyl, mono- or di-C ₁ to C ₆ alkylsulfinamoyl, optionally substituted C ₁ to C ₆ alkylcarbo Optionally substituted C ₁ to C ₆ alkylcarbonylamino, optionally substituted C ₁ to C ₆ alkoxycarbonylamino, optionally substituted phenylcarbonyl, optionally substituted heterocyclylcarbonyl, ureido, mono- or Di-C ₁ to C ₆ alkylureido, of the general formula —N (Ra) —C (Rb) = NR c, wherein Ra, R b and R c are each hydrogen or optionally substituted C ₁ to C ₆ alkyl, or Three together may form C ₃ to C ₆ cycloalkyl or heterocyclyl)

Carbamimidoyl group of formula (Rb) (Rc), wherein Ra, Rb and Rc are as defined above

Wherein R a, R b, R c and R d are each hydrogen or optionally substituted C ₁ to C ₆ alkyl, or two of them may together form C ₃ to C ₆ cycloalkyl or heterocyclyl Anidinyl groups, optionally substituted phenyl, optionally substituted heterocyclyl, C ₁ to C ₆ alkyl and optionally substituted C ₁ to C ₆ alkyl, wherein the substituents are amino, carboxyl, cyano, sulfo, halogeno, carbo Neyl, trifluoromethyl, nitro, hydroxyl, formyl, mono- (optionally substituted) C ₁ to C ₆ alkylamino, di- (optionally substituted) C ₁ to C ₆ alkylamino tertiary- (optionally substituted C ₁ to C ₆ alkylammonio, carbamoyl, mono- or di-C ₁ to C ₆ alkyl carbamoyl, carbamoyloxy, mono- or di-C ₁ to C ₆ alkyl carbamoyloxy, N (Ra) -C (Rb) = NRc, wherein Ra, Rb and Rc are each hydrogen or optionally substituted C ₁ to C ₆ alkyl, or Two adjacent ones together may form C ₃ to C ₆ cycloalkyl or heterocyclyl), and a general formula

The carbamimidoyl group, wherein Ra, Rb and Rc are as defined above.

In general, an infectious disease is caused by a pathogenic microorganism that is fixed and grows in a specific part of a living body and causes a local reaction when the microorganism accidentally penetrates a living body or the like. One effective treatment for infectious diseases is the administration of antibiotics to patients with these diseases. Commonly used effective antibiotics include penicillin and cephalosporin antibiotics having a broad antimicrobial spectrum, which are widely used to treat various kinds of infectious diseases and have excellent effects.

Thus, in most cases the pathogenic microorganisms are eradicated by the action of chemotherapeutic agents and the patient recovers. However, microorganisms that cause corresponding infectious diseases are often known to be resistant to drugs and therefore inevitable problems in chemotherapy when used for a long time.

As a result, antibiotics such as penicillin and cephalosporin antibiotics produce resistant bacteria after prolonged and extensive use, in terms of antimicrobial spectrum, antimicrobial activity, in vivo behavior of antibiotics, or safety. Not satisfactory enough.

Under these circumstances, an antibiotic called thienamycin, which is sensitive to bacteria resistant to penicillin and cephalosporin antibiotics, has been developed (see Japanese Patent Application Laid-Open No. 73191/76). Subsequently, studies were conducted to synthesize carbapenem derivatives and other penem derivatives having a skeleton structure similar to that of carbapenem. However, these commonly developed or synthesized carbapenems or penem antibiotics are all physiochemically unstable and are easily enzymatically degraded by enzymes such as dehydropeptidase in the kidney and also have relatively low and insufficient solubility in water. It is not satisfactory. Thus, very important and effective agents have not yet been developed.

As mentioned above, the use of antibiotics is very effective in the treatment of infectious diseases. However, the inevitable problem associated with chemotherapy using antibiotics is the difficulty of the emergence of resistant bacteria. In other words, conventionally developed antibiotics become increasingly insufficient in the antimicrobial spectrum involved by the development of resistant bacteria. This problem also occurs with penicillin and cephalosporins, which have a relatively broad antibacterial spectrum and are widely used.

Therefore, there is a great demand for the development of a novel medicament that is effective against bacteria resistant to the aforementioned antibiotics and has a broad antimicrobial spectrum, and various kinds of materials have been proposed so far. However, none of these substances meet the requirements for the novel medicaments described above.

Therefore, the main object of the present invention is to provide novel penem derivatives.

Another object of the present invention is to provide a novel penem derivative having effective physicochemical properties and effective against bacteria resistant to penicillin and cephalosporin antibiotics.

Another object of the present invention is to provide a method for producing the novel penem derivative.

Since the penem derivative of formula (I) of the present invention has an extremely broad antimicrobial spectrum, it has a strong antimicrobial activity against gram positive and negative bacteria and aerobic and anaerobic bacteria, as well as bacteria that are resistant to penicillin and cephalosporin antibiotics. Have In addition, the penem derivative of the present invention has excellent solubility in water, excellent physical and chemical stability, and is hardly decomposed by enzymes such as kidney dehydropeptidase I or β-lactamase. Therefore, the penem derivative of the present invention has a broad antibacterial spectrum and is excellent in biochemical stability and safety, and thus can be effectively used as an antibacterial agent.

In general formula (I), preferred penem derivatives or salts thereof of the present invention are compounds in which the substituents R ₁ , R ₂ and R ₃ are as follows. First, examples of preferred R ₁ are hydrogen; C ₁ to C ₆ alkyl groups which may have hydroxyl groups such as methyl, ethyl, n-propyl, isopropyl or n-butyl. Preferred R ₂ is hydrogen, straight or branched C ₁ to C ₆ alkyl (eg methyl, ethyl, isobutyl or tert-butyl), lower (C ₁ -C ₄ ) alkoxy C ₁ to C ₆ alkyl (eg Methoxymethyl or methoxyethyl) and lower aliphatic acyloxymethyl (e.g. pivaloyloxymethyl, phthalidyl, 5-C ₁ to C ₆ alkyl-1, or 3-dioxol-2-one-4-yl Methyl). In addition, the substituent R ₂ may serve as a group for protecting the carboxyl group when preparing the derivative of the present invention, and may be easily removed under mild conditions, for example, aralkyl (eg, o-nitrobenzyl, p-nitrobenzyl, benzhydryl or 2-naphthylmethyl), allyl or C _1-6 alkylsilyl (eg trimethylsilyl).

In addition, COOR ₂ may be a carboxylate anion depending on the kind of substituent R ₃ of the derivative of the present invention, which corresponds to the case of the carboxylate anion form as the counterion of R ₃ of the carboxyl group at the 3-position of penem. . For example, when R ₃ is a quaternary-nitrogen-containing heterocycle ring, the 3-position carboxyl group may be in the form of a carboxylate anion which is the counterion. In addition, when the derivative of the present invention is a salt with a strong acid, that is, when the quaternary-nitrogen counterion is an anion of a strong acid, R ₂ may be hydrogen, in which case the derivative is a betaine compound (quaternary-ammonium compound). Exhibits the same characteristics. Thus, acid addition salts of compound (I) can be represented by the general formula:

Wherein X ⁻ is an anion of an acid; The remaining substituents are as defined above.

On the other hand, when R ₃ is a basic group, COOR ₂ may be a carboxyl or carboxylate anion depending on the environmental state (especially pH) around the compound or may be in equilibrium between the two states. This means that the derivatives of the present invention may be in the so-called zwitterion form, and therefore, the compound of general formula (I) includes a compound having a zwitterion structure.

의 그룹, 일반식

의 C₃내지 C₆사이클로알케닐, 일반식

의 C₃내지 C₆사이클로알케닐 C₁내지 C₆알킬, 하기 일반식의 포화된 헤테로 사이클그룹:Preferred examples of the substituent R ₄ include (wherein m is 1,2 or 3; n is 1,2,3,4,5 or 6; v is 0,1 or 2; w is 0,1,2 or 3, Y is O, S or NH, Y ₁ , Y ₂ , Y ₃ and Y ₄ are each N, NH, O or S); C ₁ to C ₆ alkyl (eg methyl, ethyl, isopropyl and butyl), C ₂ to C ₆ alkenyl (eg vinyl, allyl, isopropenyl, 1-propenyl, 2-butenyl and 1,3 -Butadienyl), C ₂ to C ₆ alkynyl (e.g. ethynyl, propargyl and 2-butynyl), C ₃ to C ₆ cycloalkyl (e.g. cyclopropyl, cyclobutyl, cyclophenyl and cyclohexyl ), C ₃ to C ₆ cycloalkyl C ₁ to C ₆ alkyl (e.g. cyclopropylmethyl, cyclopropylethyl, cyclopropylisopropyl, cyclopentylmethyl and general formula

Group of, general formula

C ₃ to C ₆ cycloalkenyl, general formula

C ₃ to C ₆ cycloalkenyl C ₁ to C ₆ alkyl, saturated heterocyclic groups of the general formula:

Saturated heterocyclyl C ₁ to C ₆ alkyl of the general formula

Unsaturated (nonaromatic) heterocyclyl groups of the general formula:

Unsaturated (nonaromatic) heterocyclyl C ₁ to C ₆ alkyl of the general formula:

Unsaturated (aromatic) heterocyclyl groups of the general formula:

Unsaturated (aromatic) heterocyclyl C ₁ to C ₆ alkyl of the general formula:

의 헤테로사이클릴리덴 C₁내지 C₆알킬,General formula

Heterocyclylidene C ₁ to C ₆ alkyl,

의 사이클로알킬리덴 C₁내지 C₆알킬.General formula

Cycloalkylidene C ₁ to C ₆ alkyl.

Preferred examples of the penem derivative of the present invention are those in which the substituent R ₃ is a general formula

중에서 선택된 1개의 치환기이고;

1 substituent selected from;

Wherein A is nitrogen or carbon; R ₄ is as defined above, more preferably R ₄ is

Is selected from.

As penem derivatives of the present invention, compounds having the following bicycloheterocycle group (R ₃ ), wherein m is 1 are preferred:

In the penem derivative of general formula (I), the compound having a substituent R ₃ containing an asymmetric carbon (s) includes stereoisomers. For example, if R ₃ has one asymmetric carbon atom, two isomers must be present. One of these will then be referred to as "isomer A" and the other as "isomer B". In addition, when R ₃ has an asymmetric carbon atom and a substituent R ₄ , regioisomers other than stereoisomers may be present. In this case, they are represented by "isomer A" through "isomer D" and the like.

In general formula (I), the stereoconfiguration of the 5-position carbon atom is preferably in the R-configuration corresponding to the coordination of natural penicillin, and preferred examples of the substituent R ₁ refer to hydrogen and 1-hydroxy-ethyl group. can do. Most preferably, as in the case of thienamycin, the stereoconfiguration of the 8-position carbon atom to which the hydroxyl group is attached is the R-configuration and the 6-position carbon atom to which R ₁ such as 1-hydroxyethyl group is bonded. Stereoconfiguration is a compound of S-configuration.

In addition, the most preferred substituent R ₂ is hydrogen or anionic charge, and preferred examples of metabolizable ester residues are pivaloyloxymethyl, phthalidyl, 5-methyl-1,3-dioxol-2-one-4-yl Methyl and the like. When preparing the same compound, it is preferable that compound (I) uses a group such as p-nitrobenzyl or allyl as the protecting group for the carboxyl group.

In addition, the compounds of the present invention and certain intermediates thereof may be in tautomeric forms. Therefore, while the structural formulas described herein are shown to not include such compounds, such tautomeric forms are included within the scope of the present invention. Preferred examples of R ₃ may refer to the following compounds:

Thus, the most preferred penem derivatives of the present invention are compounds of the general formulas (la), (lb) and (lc):

Wherein R ₂ is H or COOR ₂ is a carboxylate anion; R ₄ is —CH ₃ , —C ₂ H ₅ , —CH ₂ CN, —CH ₂ , —C≡CH, —CH ₂ , — CO-CH ₃ or -CH ₂ -CO-NH ₂ )

(Wherein R ₂ is hydrogen or a carboxylate anion as COOR ₂ ; R ₄ is a methyl group)

Salts of the compounds of the general formula (I) of the present invention include non-toxic salts of carboxylic acid derivatives (in formula (I), COOR ₂ is COOH), for example metal salts such as sodium, potassium, aluminum, magnesium salts; Salts with nontoxic amines such as ammonium salt, triethylamine, procaine, benzylamine and penicillin and cephalosporin and other amines used to form salts. Most preferred salts are sodium salts and potassium salts.

On the other hand, the penem derivatives of the present invention also contain basic groups, for example inorganic acids (e.g. hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid) or organic acids (e.g. acetic acid, citric acid, succinic acid, ascorbic acid, methanesulfonic acid) Pharmaceutically acceptable acid addition salts of the family may also be present. The compounds of formula (1) of the present invention may be in solvated forms to varying degrees, for example hydrates, which are also within the scope of the present invention.

The compounds of the present invention may be mixed with pharmaceutical carriers, stabilizers, solubilizers, auxiliaries which are commonly used, according to known techniques for preparing formulations.

The pharmaceutical preparation of the present invention can be administered by various methods such as oral administration, intravenous injection, intramuscular injection or parenteral administration. The pharmaceutical formulation of the present invention is preferably in a dosage form appropriate for the particular method of administration. Therefore, the pharmaceutical preparations of the present invention may be in the form of tablets, pills, capsules, granules, injections, suppositories and the like known in the art. In general, the daily dose of a compound of the invention is 250 mg to 3,000 mg in adults, which is administered in divided doses. However, the dose may vary depending on various factors such as age, sex or the condition to be treated.

For the compound of the present invention, its antimicrobial activity is measured by Muller-Hinton broth broth dilution method in which 10 ⁵ cells / ml of bacteria are incubated at 37 ° C. for 18 hours. The results obtained are summarized in Table I.

Table I: Minimum Inhibitory Concentration (MIC: ㎍ / mL)

A: (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [2,1-c] -1,2,4-tria Sol-6-yl) thio] 2-phenem-3-carboxylic acid (isomers A, B,).

B: (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-2-methyl-5H-pyrrolo [2, 1-c] imidazolium- 7-yl) thio] 2-phenem-3-carboxylate (isomer A).

Sch 34343: A compound of the following general formula developed and marketed by Schering Company, known as an effective penem compound and reported to be excellent in clinical trials.

The penem derivative of the present invention can be easily and effectively prepared according to the method described in the following scheme.

(i) Stage 1

It can be easily prepared according to the known method of starting material (IV) which is one of the penem derivatives (see, for example, J. Am. Chem. Soc., 1982, 104, 6138) or a method for improving the same. In formula (IV), R ₁ is as defined above, R ' ₂ is an ester residue as defined for R ₂ , and R ₅ is an organic group such as alkyl, preferably lower alkyl (eg : Ethyl, n-propyl, isopropyl), aryl (eg phenyl, tolyl) or aralkyl (eg benzyl, methoxybenzyl, chlorobenzyl, phenethyl).

The phenem derivative (IV) is oxidized in an appropriate medium with an oxidizing agent such as perbenzoic acid, m-chloro-perbenzoic acid, peracetic acid, hydrogen peroxide, selenium dioxide, ozone or sodium metaperic acid, preferably m-chloro-febenzoic acid. A sulfoxide derivative (II) is obtained in high yield (in formula (II), the curved arrow attached to the sulfur atom means that the resulting compound consists of a mixture of stereoisomers). The resulting sulfoxide derivatives consisting of mixed isomers can be suitably used in the subsequent second step without separating the isomers. Solvents that can be used in this reaction include halogenated hydrocarbons (e.g. dichloromethane, chloroform, carbon tetrachloride), alcohols (e.g. methanol, ethanol), ketones (e.g. acetone, methyl acetylketone), acetic acid, pyridine, N, N- Dimethylformamide (hereinafter referred to as DMF), acetamide, dimethylsulfoxide (hereinafter referred to as DMSO), water, phosphate buffers and mixtures thereof and the like. Examples of preferred solvents include solvents that do not adversely affect the reactants or reaction products. The first step can be carried out at -50 to 50 ° C, but preferably at a milder temperature such as -30 ° C to room temperature. In general, the first stage lasts for 5 minutes to 4 hours, but generally 30 minutes to 1 hour is sufficient.

(ii) second stage

In the second step, the sulfoxide derivative (II) obtained in the first step is reacted with a thiol compound (III) (wherein R ₃ is as defined above) or an acid addition salt or reactive derivative thereof to perform a substitution reaction. In the second step, organic solvents such as DMF, DMSO, tetrahydrofuran (hereinafter referred to as THF), hexamethylphosphotriamide (hereinafter referred to as HMPA) and mixtures thereof that do not affect the reactants and products are used as reaction medium. Can be used. In general, the reaction temperature is -50 ° C to room temperature, preferably -30 ° C to 0 ° C. In general, the reaction time is 15 minutes to 2 hours, in particular 30 minutes to 1 hour.

The reactivity of the thiol compound (III) can be improved by coexisting bases in the reaction, and therefore the reaction is carried out with good speed and yield. However, of course the reaction is carried out in the absence of base. Such bases include alkylamines (e.g. triethylamine, diisopropylethylamine) '1,8-diazabicyclo [5,4,0] -7-undecene (hereinafter referred to as DBU), N-methylmor Cycloaliphatic amines such as poline; Inorganic bases such as hydroxides or carbonates of sodium and potassium; Potassium tert-butoxide, sodium methoxide and the like; Sodium amide, sodium hydride and the like, of which it is preferable to use diisopropylamine and DBU.

Examples of the reactive derivative of the thiol compound (III) include thiolate compounds of the following general formula (III ').

MS-R ₂ (III ′)

Wherein M is an alkali metal; R ₃ is as defined above. In this substitution reaction, the thiol compound of general formula (III) or acid addition salt or reactive derivative thereof is generally used in 1 to 3 equivalents, preferably 1 to 2 equivalents relative to the sulfoxide derivative (II). On the other hand, the base is preferably used in the same amount as the thiol compound (III). When the thiol compound (III) is an acid addition salt thereof, excellent results can be expected by using an excess of base such that the added acid can be neutralized.

The product resulting from this substitution can be separated according to conventional workup methods.

(iii) Third stage

If the substituted product has a protecting group, this group can be removed as the case may be. The protecting group can be removed by reductive decomposition by hydrolysis, hydrogenation or chemical reduction with acid, base or enzyme.

When R ₂ in the formula (I) is an ester residue, in particular p-nitrobenzyl, benzhydryl, 2-naphthylmethyl, or the like, the protecting group is catalytically reduced using palladium on carbon, platinum oxide or another known catalyst. To be effectively removed to obtain a penem derivative of general formula (I), wherein COOR ₂ is a carboxyl group or a carboxylate anion. In the second step, a reaction solvent such as dioxane, THF, water, a buffer or a mixture thereof, in particular a water-containing THF, a water-containing dioxane, a phosphate buffer-THF mixture, and the like can be used, and the reaction is 1 to 4 atm. Under a hydrogen pressure of 0 to 50 DEG C, preferably at 10 to 30 DEG C for 30 minutes to 16 hours, preferably for 1 to 4 hours. When R ₂ in formula (I) is a p-nitrobenzyl group, the removal of the protecting group is carried out by reacting the product with aqueous ammonium chloride solution and iron powder in THF, dioxane or the like in an aqueous solvent; When R ₂ is an allyl group, the protecting group can be removed by reacting the product with tetrakistriphenylphosphine palladium (O), triphenylphosphine and 2-ethylhexanoic acid in an aprotic solvent such as THF, methylene chloride, etc. There is; When R ₂ is a 2, 2, 2-trichloroethyl group, the protecting group can also reduce the product to zinc powder to remove, whereby the desired compound can be prepared in which COOR ₂ is a carboxyl or carboxylate anion.

Some of the products resulting from the substitution reaction [step (ii)] can sometimes be difficult to separate from each other because of their properties. Therefore, to obtain a compound of formula (I) wherein COOR ₂ is a carboxyl or carboxylate anion, the protecting group is removed in the same reactor as used to carry out the reaction without isolating the intermediate product resulting from the substitution reaction, or Excellent results can often be achieved by applying to the simple post-treatment currently used and then removing the protection group. This is a particularly simple process and results in good yield and quality of the product obtained. In addition, when using this method, it can be produced on a large scale as a target product without a complicated process.

As mentioned above, the product of the third stage may further introduce a substituent R ₄ if necessary to obtain a penem derivative (I) in which R ₃ is a quaternary nitrogen-containing heterocycle group. Introduction of R ₄ is conventional, such as alkylation methods using conventional alkylating agents such as, for example, C _1-6 alkyl halides (eg methyl iodide, ethyl iodide), bromofluoroethane, dimethylsulfate or diethylsulfate. Phosphorus technology and other identical techniques using, for example, the corresponding halides of R ₄ . In addition, examples of preferred solvents for the alkylation reaction are acetone, acetonitrile, THF, dioxane or mixtures thereof. However, the compound having a substituent R ₄ can be prepared by a process other than the third step using the raw material HS-R ₃ (III), wherein R ₄ has already been introduced.

When R ₃ in the general formula (I) is a quaternary nitrogen-containing heterocycle group, the desired compound can be obtained in good yield by alkylating the product of step (ii) and then removing the protecting group as necessary.

The desired compound of formula (I) prepared in this way can be isolated and purified according to conventional techniques. That is, for example, the product can be extracted, concentrated and separated, and then purified by methods such as recrystallization, reprecipitation and chromatography. In general, compound (I) can be crystallized and purified. To this end, it is effective or desirable to convert compound (I) to salts thereof. In this case, the salt must be a nontoxic acid addition salt. The product is first purified by crystallization as a toxic salt, followed by removal of the acid or by replacing the toxic salt with a pharmaceutically acceptable salt (which can be done by exchanging counter ions in the case of quaternary ammonium salts) to prepare the desired compound of high purity. Can be.

Esters that can be metabolized in vivo are methods commonly used to prepare esters of penicillin or cephalosporin, characterized by esterifying compound (I) in which the substituent COOR ₂ is a carboxyl or carboxylate anion] It can manufacture according to this.

In the present invention, the thiol compound of general formula (III) is also a novel compound and is useful as an intermediate for preparing the penem derivative of general formula (I). Therefore, such intermediate compounds should be considered within the scope of the present invention.

The invention will be explained in more detail through the following examples.

In the following examples and reference examples, the following abbreviations are used: PMB (p-nitrobenzyl), PMB (p-methoxybenzyl), Me (methyl), Et (ethyl), Ar (phenyl), Ts (p-toluenesulfonic acid residues), and BOC (tert-butoxycarbonyl).

Example 1

(5R, 6S, 8R) -2 [(6,7-dihydro-5H-pyrrolo [2,1-c] imidazol-7-yl) thio] -6- (1-hydroxyethyl) -2 -Synthesis of Isomers A and B of Penem-3-carboxylic Acid

(1) p-nitrobenzyl (5R, 6S, 8R) -2 [(6,7-dihydro-5H-pyrrolo [2,1-c] imidazol-7-yl) thio] -6- (1 Preparation of -Hydroxyethyl) -2-phenem-3-carboxylate

256 mg of p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -2-phenem-3-carboxylate are dissolved in 2 ml of DMF to -40 ° C. Cool. Then, to the obtained solution, 6, 7-dihydro-7-mercapto-5H-pyrrolo [1,2-c] imidazole trifluoromethane sulfonate in 2 ml of DMF (in Reference Example 1 below) 440 mg) and 0.54 ml of diisopropylethylamine are added and stirred for 30 minutes. To this reaction solution, 100 ml of ethyl acetate was added, washed with water, washed with saturated aqueous sodium chloride solution, dried over Na ₂ SO ₄ , and concentrated in vacuo. The obtained residue was purified by chromatography using a column packed with 10 g of silica gel (eluent: chloroform / methanol compound; 96: 4v / v) to give a yellow oil as the target compound (sole: 290 mg).

), 7.48(1H,

), 7.57(2H, d, J=9Hz, 2×ArH), 8.15(2H, d, J=9Hz, 2×ArH)NMR δ (CDCl ₃ ) ppm: 1.37 (3H, d, J = 6Hz, -CH ₃ ), 2.40 ~ 3.00 (1H, m), 3.00 ~ 3.50 (1H, m), 3.70 ~ 3.90 (1H, m, C ₆ -H), 4.00-4.40 (3H, m), 4.70-4.90 (1H, m), 5.30 (2H, ABq, J = 14 Hz, -OCH ₂ Ar), 5.78 (1H, brs, C ₅ -H) , 6.96 and 6.99 (1H,

), 7.48 (1 H,

), 7.57 (2H, d, J = 9 Hz, 2 x ArH), 8.15 (2H, d, J = 9 Hz, 2 x ArH)

(2) (5R, 6S, 8R) -2 [(6,7-dihydro-5H-pyrrolo [2,1-c] imidazol-7-yl) thio] -6- (1-hydroxyethyl ) Synthesis of 2-phenem-3-carboxylic acid (isomers A and B)

The compound (145 mg) obtained in the step (1) was dissolved in a mixture of 10 ml of THF and 10 ml of phosphate buffer (pH 7), 150 mg of carbon containing 10% of palladium was added thereto, and the mixture was heated at 1 atmosphere of hydrogen for 2 hours. Catalytic reduction. After filtration of the catalyst, the filtrate and washings are washed with ether. The aqueous phase is concentrated to about 20 mL volume and purified as column chromatography (column size: 20 x 300 mm) packed with diion HP-20. The fraction eluted with water was discarded and the fraction eluted with THF-water 5% (UV absorption band λ max = 324 mm) was collected and concentrated in vacuo. The obtained concentrate was subjected to high performance liquid chromatography (HPLC) [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent: 5% acetonitrile-water; Outflow rate: 5 ml / min], the fraction containing the target compound corresponding to the retention time of 10 minutes and 13.5 minutes was recovered, evaporated in vacuo and lyophilized to obtain the pale yellow powder of the target compound.

Isomer A: Yield 19 mg.

IR (KBr Disk) cm ^-1 : 1770,1580

UV λ max (H ₂ O) nm: 250 (sh), 324

NMR δ (D ₂ O) ppm: 1.39 (3H, d, J = 6Hz: -CH ₃ ), 2.6 ~ 3.05 (1H, m), 3.1 ~ 3.6 (1H, m), 4.04 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.19-4.63 (3H, m), 4.80 (DOH), 4.99-5.17 (1H, m), 5.81 (1H, d, J = 2 Hz, C ₅ -H), 7.44 (1H, s, imidazole-H), 8.61 (1H, s, imidazole-H).

HPLC (retention time): 10 minutes.

Isomer B: Yield 22 mg

IR (KBr Disk) cm ^-1 : 1770,1580

UV λ max (H ₂ O) nm: 258, 324

NMR δ (D ₂ O) ppm: 1.39 (3H, d, J = 6Hz, -CH ₃ ), 2.60 ~ 3.00 (1H, m), 3.05 ~ 3.51 (1H, m), 4.02 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.17 to 4.63 (3H, m), 4.80 (DOH), 4.97 to 5.13 (1H, m), 5.81 (1H, d, J = 2 Hz, C ₅ -H), 7.48 (1H, s, imidazole-H), 8.63 (1H, s, imidazole-H).

HPLC (retention time): 13.5 min.

Example 2

(5R, 6S, 8R) -2 [(6,7-dihydro-5H-pyrrolo [2,1-c] imidazolium-7-yl) thio] -6- (1-hydroxyethyl)- Synthesis of 2-phenem-3-carboxylate (isomers A and B)

The compound (140 mg) obtained in the step (1) of Example 1 was dissolved in a mixture of THF (3 mL) and acetone (10 mL), 0.37 mL of methyl iodide was added to the solution, and the mixture was left at 5 ° C. for 30 hours. . The reaction solution is evaporated under reduced pressure, the residue is washed with ether and dried. The powder obtained was dissolved in a mixed solvent of THF (10 mL) and water (10 mL), and then 2.5 g of ammonium chloride and 1.24 g of iron (100 mesh) were added with stirring under ice-cooling, and vigorously at this temperature for 1 hour. Stir.

The insoluble material formed is filtered off and the filtrate is washed with ethyl acetate. The aqueous phase is evaporated to about 20 mL volume and purified by column chromatography using Diion HP-20 (20 × 400 mm). After removing the fraction eluted with 150 ml of water, the fraction eluted with 5% THF-water was collected, concentrated under reduced pressure, and then purified by HPLC [carrier: nucleosil 7C ₁₈ (10 × 100 mm); Solvent: 7% acetonitrile-water; Effluent rate: 4 ml / min] to recover the fraction containing the target compound in the retention time 11 minutes and 13.5 minutes respectively.

In this way, evaporation and lyophilization under vacuum yields the above pale yellow powder as the target compound.

Isomer A: Yield 14 mg.

IR (KBr Disk) cm ^-1 : 1760,1580

UV λ max (H ₂ O) nm: 250 (sh), 325

NMR δ (D ₂ O) ppm: 1.38 (3H, d, J = 6Hz: -CH ₃ ), 2.63 ~ 3.02 (1H, m), 3.08 ~ 3.55 (1H, m), 3.96 (3H, s, -NCH ₃ ), 4.04 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.16-4.60 (3H, m), 4.80 (DOH), 4.96-5.13 (1H, m), 5.79 (1H, d, J = 2 Hz, C ₅ -H), 7.47 (1H, s, imidazole-H), 8.70 (1H, s, imidazole-H).

HPLC (retention time): 11 min.

Isomer B: Yield 17 mg

IR (KBr Disk) cm ^-1 : 1760,1595

UV λ max (H ₂ O) nm: 260, 325

NMR δ (D ₂ O) ppm: 1.38 (3H, d, J = 6Hz, -CH ₃ ), 2.55 ~ 2.90 (1H, m), 3.00 ~ 3.50 (1H, m), 3.96 (3H, s, -NCH ₃ ), 4.02 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.16-4.60 (3H, m), 4.80 (DOH), 4.95-5.22 (1H, m), 5.80 (1H, d, J = 2 Hz, C ₅ -H), 7.51 (1H, s, imidazole-H), 8.70 (1H, s, imidazole-H).

HPLC (retention time): 13.5 min.

Example 3

(5R, 6S, 8R) -2 [(6,7-dihydro-2- (2-oxopropyl) -5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] -6 Synthesis of-(1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

The product (200 mg) obtained in step (1) of Example 1 was dissolved in acetone (15 mL), and 0.69 mL of bromoacetone was added, followed by stirring for 4 hours. After evaporating the solvent, the obtained residue is washed with ether and dried. The obtained solid is dissolved in 25 ml of a 50% THF-water mixture, mixed with 3.6 g of ammonium chloride and 1.8 g of iron, ice-cooled and stirred for 1.5 h.

After insoluble material is filtered off, the solution is evaporated to about 5 ml under reduced pressure. The concentrate was passed through a column packed with Diaion HP-20 (manufactured and sold by Mitsubishi Chemical Corporation), and the fraction eluted with 5%, THF-water was purified again by HPLC to give the desired compound.

HPLC Conditions

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Flow rate: 3.65 ml / min

Solvent: 7% Acetonitrile-water

Isomer A: Yield 45 mg.

IR (KBr disk) cm ^-1 : 3410, 2940, 1770, 1590, 1360

UV λ max (H ₂ O) nm: 248 (sh), 325

NMR δ (D ₂ O) ppm: 1.33 (3H, d, J = 6.6 Hz: -CH ₃ ), 2.36 (3H, s), 2.6-3.0 (1H, m), 3.0-3.6 (1H, m), 3.99 (1H, dd, J = 1, 4 Hz, 6.1 Hz, C ₆ -H), 4.29 (1H, t, J-6.1 Hz, C ₈ -H), 4.3-4.8 (2H, m), 4.74 (DOH ), 4.8 to 5.1 (1H, m), 5.35 (2H, s), 5.73 (1H, d, J = 1.4 Hz, C ₅ -H), 7.37 (1H, s, side chain C ₁ -H), 8.67 ( 1H, s, side chain C ₃ -H).

HPLC (retention time): 19 minutes.

Isomer B: Yield 54 mg

IR (KBr Disk) cm ^-1 : 3400, 2960, 1765,1590, 1360

UV λ max (H ₂ O) nm: 259 (sh), 324

NMR δ (D ₂ O) ppm: 1.33 (3H, d, J = 6.3Hz, -CH ₃ ), 2.37 (3H, s), 2.5 ~ 2.9 (1H, m), 2.9 ~ 3.5 (1H, m), 3.97 (1H, dd, J = 1.4 Hz, 6.1 Hz, C ₆ -H), 4.29 (1H, t, J = 6.1 Hz, C ₈ -H), 4.3-4.7 (2H, m), 4.74 (DOH) , 4.9 to 5.3 (1H, m), 5.36 (2H, s), 5.73 (1H, d, J = 1.4 Hz), 7.41 (1H, s, side chain C ₁ -H), 8.69 (1H, s, side chain C ₃ -H).

HPLC (retention time): 24 min.

Example 4

(5R, 6S, 8R) -2 [(6,7-dihydro-2- (cyclopropylcarbonyl) -5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] -6 Synthesis of-(1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

The product (195 mg) obtained by the method of Example 1- (1) is dissolved in acetone (15 mL). To this solution was added 1.45 g of cyclopropyl carbonyl iodide (see J. Am. Chem. Soc., 1962, 86, 2247) and stirred for 24 hours, followed by 0.73 g of cyclopropylcarbonyl iodide Stir for 24 hours. After filtration off the solvent, the residue is washed with ether and dried. The obtained solid is dissolved in 25 ml of 50% THF-water, 3.5 g of ammonium chloride and 1.7 g of iron are added, followed by stirring for 1.5 hours under ice-cooling. The solution is filtered off to insoluble material and then concentrated to about 5 ml under reduced pressure. The concentrate was passed through a column packed with Diaion HP-20, and the fractions eluted with 5% THF-water were again purified by HPLC (carrier: nucleosil 7C ₁₈ ; solvent: 10% acetonitrile; effluent rate: 4.75 mL / min). ) To give 9 mg of the target compound.

IR (KBr disk) cm ^-1 : 3360, 3110, 2950, 1765, 1580, 1440, 1350, 1280, 1120.

UV λ max (H ₂ O) nm: 250 (sh), 325

NMR δ (D ₂ O) ppm: 0.3 ~ 0.6 (2H, m), 0.6 ~ 1.0 (2H, m), 1.2 ~ 1.5 (1H, m) 1.33 (3H, d, J = 6.3Hz, -CH ₃ ) , 2.4 ~ 3.0 (1H, m), 3.0 ~ 3.5 (1H, m), 3.98 (1H, d, J = 6.1Hz, C ₆ -H), 4.07 (2H, d, J = 7.4Hz), 4.3 ~ 4.7 (2H, m), 4.74 (DOH), 4.9-5.2 (1H, m), 5.73 (1H, brs, C ₅ -H), 7.52 (0.5H, s, branched chain C ₁ -H), 7.56 (0.5) H, s, side chain C ₁ -H), 8.76 (1H, s, side chain C ₃ -H).

HPLC (holding time): 20 minutes

Example 5

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(2-cyanomethyl-6,7-dihydro-5H-pyrrolo [1,2-c] imidazolium- Synthesis of 7-yl) -thio] -2-phenem-carboxylate (isomers A and B):

Bromoacetonitrile 20eq. Is added instead of bromoacetone and the method of Example 3 is repeated except that the reaction is continued at 0 to 5 ° C for 20 hours. Thereafter, the same post-treatment with the following HPLC conditions is performed to obtain the target compound.

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 7% Acetonitrile-water

Flow rate: 5 ml / min

Isomer A: Yield 39%

IR (KBr Disk) cm ^-1 : 1765, 1590

HPLC (retention time): 9 minutes.

Isomer B: Yield 43%

IR (KBr Disk) cm ^-1 : 1765, 1595

HPLC (holding time): 11.5 minutes

Example 6

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(2-carboxylatemethyl-6,7-dihydro-5H-pyrrolo [1,2-c] imidazolium- Synthesis of 7-yl) -thio] -2-phenem-3-carboxylate (isomers A and B):

P-nitrobenzyl ester of bromoacetoic acid 5eq. Was added instead of bromoacetone, and the reaction was carried out at 10 ° C. for 3 days. Obtain the desired compound:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 5% Methanol-Water

Flow rate: 5 ml / min

Isomer A: Yield 19%

UV lambda max (H ₂ O) nm: 250 (sh), 325

HPLC (retention time): 7.5 min.

Isomer B: Yield 20%

UVλ max (H ₂ O) nm: 255 (sh), 325

HPLC (holding time): 8.5 minutes

Example 7

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(2-allyl-6,7-dihydro-5H-pyrrolo [1,2-c] imidazolium-7- Synthesis of yl) thio] -2-phenem-3-carboxylate (mixture of isomers A and B):

Allyl bromide 20eq. Was added instead of bromoacetone, and the reaction of Example 3 was repeated except that the reaction was continued at 0 to 5 ° C. for 20 hours, and subjected to the same post-treatment under the following HPLC conditions to obtain the target compound. To obtain:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 10% acetonitrile-water

Flow rate: 6 ml / min

Isomer Mixtures A and B: Yield 34%

UV lambda max (H ₂ O) nm: 253 (sh), 324

HPLC (hold time): 9.7 and 10.2 min.

Example 8

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-2-propargyl-5H-pyrrolo [1,2-c] imidazolium- Synthesis of 7-yl) thio] -2-phenem-3-carboxylate (isomers A and B):

Except for continuing reaction at room temperature for 17 hours using propargyl bromide 20eq. Instead of bromoacetone, following the same work up to the following HPLC conditions to afford the desired compound:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 10% acetonitrile-water

Flow rate: 6 ml / min

Isomer A: Yield 20%

UV lambda max (H ₂ O) nm: 250 (sh), 324

HPLC (holding time): 7.6 minutes

Isomer B: Yield 23%

UVλ max (H ₂ O) nm: 257, 324

HPLC (holding time): 8.6 minutes

Example 9

(5R, 6S, 8R) -2- (6,7-dihydro-2- (methoxycarbonylmethyl) -5H-pyrrolo [1,2-c] imidazolium-7-yl) thio]- Synthesis of 6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

The product (150 mg) obtained according to the method of Example 1- (1) is dissolved in acetone (12 ml), 0.57 ml of methyl bromoacetate is added and stirred for 96 hours. After evaporating the solvent, the obtained residue is washed with ether and dried. The obtained solid is dissolved in 50% THF-water (23 mL), 3.1 g ammonium chloride and 1.6 g iron are added and stirred under ice cooling for 2 hours.

After insoluble material is filtered off, the filtrate is evaporated to about 5 ml volume. The concentrate was passed through a column packed with Diaion HP-20, and then the fractions eluted with 5% THF-water were purified by HPLC to afford the desired compound. The conditions of HPLC are as follows.

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 10% acetonitrile-water

Flow rate: 3.65 ml / min

Isomer A: Yield 7 mg

IR (KBr Disk) cm ^-1 : 3400, 1760, 1590, 1360

UV λ max (H ₂ O) nm: 248 (sh), 322

NMR δ (D ₂ O) ppm: 1.43 (3H, d, J = 6Hz: -CH ₃ ), 2.5 ~ 3.0 (1H, m), 3.0 ~ 3.6 (1H, m), 3.86 (3H; s; -COOCH ₃ ), 3.95 (1H; d; J = 6 Hz; C ₆ -H), 4.29 (1H, t, J-6 Hz, C ₈ -H), 4.3-4.7 (2H; m), 4.74 (DOH), 4.9 5.2 (1H; m), 5.20 (2H; s), 5.83 (1H; brs, C ₅ -H), 7.56 (1H; s; side chain C ₁ -H), 8.81 (1H; s; side chain C _3- H).

HPLC (retention time): 9 minutes.

Isomer B: Yield 14 mg

IR (KBr Disk) cm _-1 : 3400, 1760, 1590, 1370

UV λ max (H ₂ O) nm: 259 (sh), 325

NMR δ (D ₂ O) ppm: 1.33 (3H; d; J = 6.3 Hz; -CH ₃ ), 2.4-3.0 (1H, m), 3.0-3.5 (1H; m), 3.86 (3H; s; COOCH ₃ ), 3.96 (1H; d; J = 6.3 Hz; C ₆ -H), 4.29 (1H; t; J = 6.3 Hz; C ₈ -H), 4.4-4.7 (2H; m), 4.74 (DOH ), 4.9 to 5.2 (1H; m), 5.20 (2H; s), 5.73 (1H; brs; C ₅ -H), 7.55 (1H; s; side chain C ₁ -H), 8.81 (1H; s; side chain C ₃ -H).

HPLC (retention time): 16 minutes.

Example 10

(5R, 6S, 8R) -2-[(6,7-dihydro-2- (carbamoylmethyl) -5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] -6 Synthesis of-(1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

The penem derivative (196 mg) obtained according to the method described in Example 1- (1) is dissolved in acetone (15 ml), iodo acetamide (1.48 g) is added and stirred for 23 hours. The solvent is evaporated off under reduced pressure, and the obtained residue is washed with ether and dried.

The obtained solid is dissolved in 50% THF-water, ammonium chloride (3.6 g) and iron (1.8 g) are added and stirred under ice cooling for 1.5 hours. After insoluble matters were filtered off from the reaction solution, the filtrate was concentrated to about 5 mL and passed through a column packed with Diion HP-20. Fractions eluted with 5% THF-water are mixed and purified again by HPLC under the following conditions.

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 7% Acetonitrile-water

Flow rate: 3.65 ml / min

In this way, the desired isomer is prepared.

Isomer A: Yield 45 mg

IR (KBr Disk) cm ^-1 : 3380, 1760, 1680, 1580, 1360, 1290, 1120

UV λ max (H ₂ O) nm: 248 (sh), 326

NMR δ (D ₂ O) ppm: 1.31 (3H; d; J = 6.4 Hz: -CH ₃ ), 2.5-3.0 (1H; m), 3.0-3.6 (1H; m), 3.99 (1H; dd; J = 1.3 Hz; 5.9 Hz; C ₆ -H), 4.1-4.6 (2H; m), 4.26 (1H, t; J = 5.9 Hz; C ₈ -H), 4.74 (DOH), 4.9-5.2 (1H; m), 5.08 (2H; s), 5.72 (1H; d; J = 1.3 Hz; C ₅ -H), 7.46 (1H; s; side chain C ₁ -H), 8.77 (1H; s; side chain C _3- H).

HPLC (retention time): 11 min.

Isomer B: Yield 57 mg

IR (KBr Disk) cm ^-1 : 3380, 1765, 1695, 1590, 1370, 1295, 1130

UV λ max (H ₂ O) nm: 256 (sh), 324

NMR δ (D ₂ O) ppm: 1.30 (3H; d; J = 6.4Hz; -CH ₃ ), 2.4 ~ 2.9 (1H; m), 2.9 ~ 3.5 (1H; m), 3.95 (1H; dd; J = 1.5 Hz, 5.9 Hz; C ₆ -H), 4.1-4.6 (2H; m), 4.26 (1H; t; J = 5.9 Hz; C ₈ -H), 4.74 (DOH), 4.9-5.2 (1H; m), 5.09 (2H; s), 5.71 (1H; d; J = 1.5 Hz; C ₅ -H), 7.50 (1H; s; C ₁ -H), 8.78 (1H; s; side chain C ₃ -H ).

HPLC (retention time): 16 minutes.

Example 11

(5R, 6S, 8R) -2-[(6,7-dihydro-2-benzyl-5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] -6- (1- Hydroxyethyl) -2-phenem-3-carboxylate (mixture of isomers A and B):

The process of Example 3 was carried out except that benzyl bromide 20eq. Was used in place of bromoacetone and the reaction was continued for 18 hours so that HPLC was carried out under the following conditions The desired compound (yield: 60%) after the post-treatment is obtained:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 15% acetonitrile-water

Flow rate: 5.9ml / min

IR (KBr disk) cm ^-1 : 3420, 1770, 1590, 1450, 1360, 1285

UV lambda max (H ₂ O) nm: 253 (sh), 325

NMR δ (D ₂ O) ppm: 1.31 (3H; d; J = 6 Hz; CH ₃ ), 2.5-3.0 (1H; m), 3.0-3.5 (1H; m), 3.75 and 3.89 (0.5H, respectively); Each d; J = 5.9 Hz; C ₆ -H), 4.1-4.6 (3H; m; C ₈ -H, side chain C ₅ -H), 4.74 (DOH), 4.9-5.1 (1H; m), 5.41 (2H; s), 5.49 and 5.60 (0.5H each; brs; C ₅ -H), 7.48 (6H; brs; side chain C ₁ -H, Ar-H), 8.78 and 8.82 (0.5H; respectively) S; side chain C ₃ -H).

HPLC (holding time): 16 minutes (Isomer A)

17 minutes (Isomer B)

Example 12a

(5R, 6S, 8R) -2-[(6,7-dihydro-2-ethyl-5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] -6- (1- Synthesis of hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

p-nitrobenzyl (5R, 6S, 8R) -2-[(6,7-dihydro-5H-pyrrolo [1,2-c] imidazol-7-yl) thio] -6- (1-hydro Roxyethyl) -2-phenem-3-carboxylate (161 mg) is dissolved in acetone (15 mL), then 0.53 mL of ethyl iodide is added three times every 24 hours and stirred for 48 hours. The residue obtained is washed with ether and the solvent is distilled off before drying. The obtained solid is dissolved in 25 ml of 50% THF-water, 3.2 g ammonium chloride and 1.6 g iron and stirred for 70 minutes under ice cooling. The insolubles are filtered off before the filtrate is distilled to about 5 ml under reduced pressure. The concentrate was passed through column Diaion HP-20 and eluted with 5% THF-water, and purified by effective HPLC under the following conditions.

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 10% acetonitrile-water

Flow rate: 3.65 ml / min

This gives the desired compounds (isomers A and B):

Isomer A: Yield 32 mg

IR (KBr disc) cm ^-1 : 3400, 1765, 1580, 1445, 1360, 1280, 1120

UV λ max (H ₂ O) nm: 248 (sh), 324

NMR δ (D ₂ O) ppm: 1.34 (3H; d; J = 6.1Hz: CH ₃ ), 1.52 (3H; t; J = 7.2Hz), 2.6 ~ 3.0 (1H; m), 3.0 ~ 3.5 (1H m), 4.00 (1H; d; J = 5.9 Hz; C ₆ -H), 4.1-4.7 (3H; m), 4.26 (2H, q; J = 7.2 Hz), 4.74 (DOH), 4.9-5.1 (1H; s; side chain C ₅ -H), 5.75 (1H; s; C ₅ -H), 7.50 (1H; s; side chain C ₁ -H), 8.71 (1H; s; side chain C ₃ -H).

HPLC (retention time): 13 minutes.

Isomer B: Yield 44 mg

IR (KBr Disk) cm ^-1 : 3420, 1765, 1590, 1440, 1360, 1285, 1120

UV λ max (H ₂ O) nm: 258 (sh), 324

NMR δ (D ₂ O) ppm: 1.33 (3H; d; J = 6.4Hz; CH ₃ ), 1.52 (3H; t; J = 7.2Hz), 2.5 ~ 2.9 (1H; m), 2.9 ~ 3.5 (1H m), 3.97 (1H; d; J = 5.9 Hz; C ₆ -H), 4.1-4.7 (3H; m), 4.26 (2H; q; J = 7.2 Hz), 4.74 (DOH), 4.9-5.1 (1H; m), 5.77 (1H; s; C ₅ -H), 7.54 (1H; s; side chain C ₁ -H), 8.71 (1H; s; side chain C ₃ -H).

HPLC (retention time): 15 minutes.

Example 12b

(5R, 6S, 8R) -2-[(6,7-dihydro-2- (3-methoxycarbonyl-2-oxopropyl) -5H-pyrrolo [1,2-c] imidazolium- Synthesis of 7-yl) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

According to the method of Example 3, except that methyl bromoacetate 10eqs was used instead of bromoacetone and the reaction was carried out at room temperature for 5 hours, HPLC was carried out according to Example 1- (2), which was effective under the following conditions After treatment, the desired compound is obtained:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 10% acetonitrile-water

Flow rate: 6 ml / min

Isomer A: Yield 10%

UVλmas (H ₂ O) nm: 250 (sh), 325

HPLC (retention time): 14.5 min.

Isomer B: Yield 12%

UVλ max (H ₂ O) nm: 255 (sh), 324

HPLC (retention time): 18.8 min.

Example 12c

(5R, 6S, 8R) -2-[(6,7-dihydro-2- (4-fluorobenzyl) -5H-pyrrolo [1,2-c] imidazolium-7-yl) thio] A mixture of -6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B):

According to the method of Example 3, except that 4-fluorobenzylbromide 10eqs. Was used in place of bromoacetone and the reaction was carried out at room temperature for 17 hours, HPLC was carried out in Example 1- (2), which was effective under the following conditions. After treatment according to the following procedure, the desired compound is obtained:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 20% acetonitrile-water

Flow rate: 6 ml / min

Mixture of Isomers A and B: Yield 43%

UVλ max (H ₂ O) nm: 255 (sh), 324

HPLC (hold time): 14.5 min and 15 min.

Example 13

(5R, 6S, 8R) -2-[(6,7-dihydro-methyl-5H-pyrrolo [2,1-c] -1,2,4-triazol-7-yl) thio] -6 Synthesis of-(1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B and mixtures of isomers C and D):

(1) (5R, 6S, 8R) -2-[(6,7-dihydro-5H-pyrrolo [1,2-c] -1,2,4-triazol-7-yl) thio]- Synthesis of p-nitrobenzyl ester of 6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid:

The thiol derivative (361 mg) obtained by the method described in Reference Example 1 was treated according to the method of Example 1- (1) to prepare the target compound (yield: 207 mg) described above.

NMRδ (CDCl ₃ ) ppm: 1.40 (3H; d; J = 6 Hz; CH ₃ ), 2.50-3.0 (1H; m; pyrrole ring C ₆ -H), 3.0-3.6 (1H; m; pyrrole ring C _6- H), 3.6-4.5 (4H, m; pyrrole rings C ₅ -H ₂ , C ₆ -H and C ₈ -H), 4.85 (1H; dd; J = 8.4 Hz; pyrrole rings C ₇ -H), 5.25 And 5.39 (1H: ABq; J = 14 Hz; CO ₂ H ₂ Ar), 5.31 and 5.43 (1H; ABq; J = 14 Hz; CO ₂ CH ₂ Ar), 7.63 and 7.65 (1H; d; J = 9 Hz; ArH ), 8.20 (1H; s; triazole ring C ₃ -H), 8.22 and 8.24 (1H; d; J = 9 Hz; ArH)

(2) (5R, 6S, 8R) -2-[(6,7-dihydro-methyl-5H-pyrrolo [1,2-c] -1,2,4-triazol-7-yl) thio ] -6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomer A and B, mixture of isomers C and D)

The compound (340 mg) obtained in the step (1) was treated in the same manner as described in Example 2 to obtain the target compound described above. Purification is carried out according to HPLC carried out under the following conditions:

Carrier: Nucleosil 7C ₁₈ (10 × 300㎜)

Solvent: 5% acetonitrile-water

Flow rate: 3.65 ml / min

Isomer A: Yield 8 mg

IR (KBr disc) cm ^-1 : 1775, 1600

UV lambda max (H ₂ O) nm: 257, 327

NMR δ (D ₂ O) ppm: 1.33 (3H; d; J = 6Hz; CH ₃ ), 2.9-3.3 (1H; m; pyrrole ring C ₆ -H), 3.3-3.9 (1H; m; pyrrole ring C ₆ -H), 4.04 (1H; dd; J = 6, 2 Hz; C ₆ -H), 4.16 (3H; s; NCH ₃ ), 4.1-4.7 (3H; m; pyrrole rings C ₅ -H ₂ and C ₈ H), 5.18 (1H; dd; J = 10,5 Hz; pyrrole ring C ₇ -H), 5.76 (1H; d; J = 2 Hz; C ₅ -H), 8.80 (1H; s; triazole ring C ₃ -H)

HPLC (retention time): 12.1 min.

Isomer B: Yield 20 mg

IR (KBr disc) cm ^-1 : 1775, 1600

UV lambda max (H ₂ O) nm: 261, 328

NMR δ (D ₂ O) ppm: 1.33 (3H; d; J = 6Hz; CH ₃ ), 2.9-3.3 (1H; m; pyrrole ring C ₆ -H), 3.3-3.9 (1H; m; pyrrole ring C ₆ -H), 4.04 (1H; dd; J = 6, 2 Hz; C ₆ -H), 4.18 (3H; s; NCH ₃ ), 4.1-4.7 (3H; m; pyrrole rings C ₅ -H ₂ and C _8- H), 5.18 (1H; dd; J = 10.5 Hz; pyrrole ring C ₇ -H), 5.76 (1H; d; J = 2 Hz; C ₅ -H), 8.80 (1H; s; triazole ring C ₃ -H)

HPLC (retention time): 13.5 min.

Mixture of Isomers C and D: Yield 91 mg

IR (KBr disc) cm ^-1 : 1775, 1600

UV λ max (H ₂ O) nm: 253, 326

NMR δ (D ₂ O) ppm: 1.36 (3H; d; J = 6Hz; CH ₃ ), 2.6-3.1 (1H; m; pyrrole ring C ₆ -H), 3.1-3.7 (1H; m; pyrrole ring C _6- H), 4.07 (1.5H; s; NCH ₃ ), 4.08 (1.5H; s; NHC ₃ ), 4.0-4.8 (4H; m; pyrrole ring C ₅ -H ₂ , C ₆ -H, and C ₈ -H), 5.12 (1H; dd; J = 10, 5 Hz; pyrrole ring C ₇ -H), 5.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (holding time): 16.2 and 16.6 minutes

Example 14

(5R, 6S, 8R) -2-[(6,7-dihydro-3-trifluoromethyl-5H-pyrrolo [2,1-c] -1,2,4-triazol-6-yl Synthesis of Thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B)

(1) (5R, 6S, 8R) -2-[(6,7-dihydro-3-trifluoromethyl-5H-pyrrolo [2,1-c] -1,2,4-triazole- Synthesis of 6-yl) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid: A thiol derivative (231 mg) prepared according to the method described in Reference Example 2 was prepared according to Example 1-. Preparation is carried out according to the method described in (1).

NMR δ (CDCl ₃ ) ppm: 1.40 (3H; d; J = 6Hz; CH ₃ ), 3.0-3.4 (1H; m; pyrrole ring C ₇ -H), 3.4-3.7 (1H; m; pyrrole ring C ₇ -H), 3.84 (1H, m pyrrole ring C ₆ -H), 4.1-4.5 (2H; m; pyrrole ring C ₆ -H and C ₆ -H), 4.5-4.9 (2H; m; pyrrole ring C ₅ -H), 5.20 and 5.48 (2H; ABq; J = 14 Hz; CO ₂ CHAr), 7.60 (2H; d; J = 9 Hz; ArH), 8.24 (2H; d; J = 9 Hz; ArH)

(2) sodium (5R, 6S, 8R) -2-[(6,7-dihydro-3-trifluoroethyl-5H-pyrrolo [2,1-c] -1,2,4-triazole Synthesis of -6-yl) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylate (isomers A and B)

The compound is obtained by using the compound (59 mg) obtained in the above step (1) and repeating the same method as described in Example 1- (2). The product is purified by HPLC under the following conditions.

Carrier: Nucleosil 7C ₁₈ (10 × 300mm)

Solvent: 5% acetonitrile-water

Flow rate: 3.65 ml / min

Isomer A: Yield 13 mg

IR (KBr Disk) cm ^-1 : 1770, 1595

UV λ max (H ₂ O) nm: 256, 323

NMR δ (D ₂ O) ppm: 1.36 (3H; d; J = 6Hz; CH ₃ ), 3.09 (1H; dd; J = 18, 5Hz; pyrrole ring C ₇ -H), 3.68 (1H; dd; J = 18, 7 Hz; pyrrole ring C ₇ -H), 3.99 (1H; dd; J = 7, 2 Hz; C ₆ -H), 4.2-4.7 (4H; m; pyrrole ring C ₅ -H ₂ , pyrrole ring C ₆ -H and C ₈ -H), 5.76 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 15.4 min.

Isomer B: Yield 13 mg

IR (KBr disc) cm ^-1 : 1760, 1590

UVλmax (H ₂ O) nm: 252, 324

NMRδ (D ₂ O) ppm: 1.40 (3H; d; J = 6Hz; CH ₃ ), 3.28 (1H; dd; J = 18, 5Hz; pyrrole ring C ₇ -H), 3.78 (1H; dd; J = 18, 7 Hz; pyrrole ring C ₇ -H), 4.00 (1H; dd; J = 7, 2 Hz; C ₆ -H), 4.2-4.7 (4H; m; pyrrole ring C ₅ -H ₂ , pyrrole ring C ₆ -H and C ₈ -H), 5.76 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 18.6 min.

According to the same method as described above to prepare the following compound:

(a) (5R, 6S, 8R) -2-[(3-amino-6,7-dihydro-5H-pyrrolo [2,1-c] -1,2,4-triazol-7-yl Mixture of isomers A and B of thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid

HPLC (holding time): 8 minutes

HPLC conditions:

Carrier: Nucleosil 7C ₁₈ (10 × 300mm)

Solvent: 5% Methanol-Water

Flow rate: 4ml / min

IR (KBr disc) cm ^-1 : 1776

NMRδ (D ₂ O) ppm: 1.40 (3H; d; J = 6 Hz; CH ₃ ), 2.60-3.20 (and 3.20-3.65 (1H; m; -CH ₂ CH ₂ N), 3.95-4.50 (4H; m ; C ₆ -H, C ₈ -H, -CH ₂ N), 4.80 (HOD), 5.80 (1H; d; J = 2Hz; C ₅ -H)

(b) (5R, 6S, 8R) -2-[(3-chloro-6,7-dihydro-5H-pyrrolo [2,1-c] -1,2,4-triazol-7-yl A mixture of isomers A and B of) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid.

HPLC (holding time): 17 minutes

Condition :

Carrier: Nucleosil 7C ₁₈ (10 × 300mm)

Solvent: 5% acetonitrile-water

Flow rate: 4ml / min

IR (KBr disc) cm ^-1 : 1770

NMRδ (D ₂ O) ppm: 1.38 (3H; d; J = 6Hz; CH ₃ ), 2.60-3.80 (2H; m; -CH ₂ CH ₂ N), 3.85-4.40 (4H; m; C ₆ -H , C ₈ -H, -CH ₂ N), 4.80 (HOD), 5.78 (1H; d; J = 2Hz; C ₅ -H)

(c) (5R, 6S, 8R) -2-[(2-amino-6,7-dihydro-5H-phyllo [2,1-b] -1,2,4-triazol-7-yl Isomers A and B of) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid.

Isomer A:

IR (KBr disc) cm ^-1 : 1767

NMRδ (D ₂ O) ppm: 1.35 (3H; d; J = 6Hz; CH ₃ ), 2.55-3.60 (2H; m; CH ₂ CH ₂ N), 3.85-4.50 (4H; m; C ₅ -H, C ₈ -H, -CH ₂ N), 4.80 (HOD), 5.75 (1H; d; J = 2Hz; C ₅ -H)

HPLC (holding time): 12 minutes

Condition: Nucleosil 7C ₁₈ (10 × 30mm)

3% acetonitrile-water

4ml / min

Isomer B:

IR (KBr disc) cm ^-1 : 1770

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz; CH ₃ ), 2.60-3.60 (2H; m; CH ₂ CH ₂ N), 3.85-4.40 (4H; m; C ₆ -H, C ₈ -H, -CH ₂ N), 4.30 (HOD), 5.90 (1H; d; J = 2Hz; C ₅ -H)

HPLC (retention time): 13 minutes.

(d) (5R, 6S, 8R) -2-[(2-chloro-6,7-dihydro-5H-pyrrolo [2,1-b] -1,2,4-triazol-7-yl Isomers A and B of) thio] -6- (1-hydroxy) -2-phenem-3-carboxylic acid.

Isomer A:

IR (KBr disc) cm ^-1 : 1770

NMRδ (D ₂ O) ppm: 1.40 (3H; d; J = 6 Hz; CH ₃ ), 2.60-3.00 and 3.20-3.62 (1H; m; CH ₂ CH ₂ N), 4.80 (HOD), 5.78 (1H, d, J = 2 Hz; C ₅ -H)

HPLC (retention time): 14 minutes.

Condition: Nucleosil 7C ₁₈ (10 × 30mm)

5% acetonitrile-water

4ml / min

Isomer B:

IR (KBr disc) cm ^-1 : 1770

NMRδ (D ₂ O) ppm: 1.35 (3H; d; J = 6 Hz; CH ₃ ), 2.65-3.05 and 3.18-3.60 (1H; m; CH ₂ CH ₂ N), 3.85-4.55 (4H; m; C ₅ -H, C ₈ -H, -CH ₂ N), 4.80 (HOD), 5.78 (1H; d; J = 2Hz; C ₅ -H)

HPLC (retention time): 16 minutes.

(e) (5R, 6S, 8R) -2-[(6,7, -dihydro-5H-pyrrolo [2,1-c] -triazolium-6-yl) thio] -6- (1- Isomers A and B of hydroxyethyl) -2-phenem-3-carboxylate.

Isomer A:

IR (KBr disc) cm ^-1 : 3430, 1765, 1370, 1280, 1260

UVλmax (H ₂ O) nm: 249, 323

NMRδ (D ₂ O) ppm: 1.31 (3H; d; J = 6.2Hz; CH ₃ ), 3.0-3.8 (2H; m), 3.97 (1H; dd; J = 1.1Hz, 6.4Hz; C ₆ -H ), 4.27 (1H, t; J = 6.4 Hz; C ₈ -H), 4.2-5.2 (3H; m), 4.74 (DOH), 5.75 (1H: d; J = 1.1 Hz; C ₅ -H), 7.85 (1H; s; branched C ₁ -H), 9.84 (1H; s; branched C ₃ -H)

HPLC (holding time): 14 minutes

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

5% acetonitrile-water

3.65ml / min

Isomer B:

IR (KBr disc) cm ^-1 : 3420, 1770, 1580, 1370, 1280, 1260

UVλmax (H ₂ O) nm: 249, 323

NMRδ (D ₂ O) ppm: 1.31 (3H; d; J = 6.2Hz; CH ₃ ), 3.0-3.8 (2H; m), 4.01 (1H; dd; J = 1.1Hz; 6.4Hz; C ₆ -H ), 4.27 (1H; t; J = 6.4 Hz; C ₈ -H), 4.2-5.2 (3H; m), 4.74 (DOH), 5.77 (1H; d; J = 1.1 Hz; C ₅ -H), 4.86 (1H; s; branched C ₁ -H), 9.84 (1H; s; branched C ₃ -H)

HPLC (retention time): 17 minutes.

(f) (5R, 6S, 8R) -2-[(5,6-dihydro-4H-pyrrolo [1,2-c] -1,2,4-triazol-4-yl) thio]- Mixture of Isomers A and B of 6- (1-hydroxyethyl) -2-phenem-3-carboxylic acid:

IR (KBr disc) cm ^-1 : 3400, 1765, 1585, 1380

UVλ max (H ₂ O) nm: 253, 324

NMRδ (D ₂ O) ppm: 1.34 (3H; d; J = 6Hz; CH ₃ ), 2.6-3.2 (1H; m), 3.2-3.7 (1H; m), 3.8-4.1 (1H; m; C ₆ -H), 4.27 (1H; t; J = 6 Hz; C ₈ -H), 4.3-5.2 (3H; m), 4.74 (DOH), 5.72 (1H; brs, C ₅ -H), 7.75 (0.5H ; s; side chain C ₃ -H), 7.79 (0.5H; s; side chain C ₃ -H)

HPLC (holding time): 16.5 minutes

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

5% acetonitrile-water

3.65ml / min

(g) (5R, 6S, 8R) -2-[(5,6-dihydro-2-methyl-4H-pyrrolo [1,2-c] -1,2,3-triazolinium-4- Isomers A and B of yl) thio] -6- (1-hydroxyethyl) -2-phenem-3-carboxylate:

Isomer A:

IR (KBr disc) cm ^-1 : 1765

UVλ max (H ₂ O) nm: 325

NMRδ (D ₂ O) ppm: 1.34 (3H; d; J = 5.9 Hz; CH ₃ ), 2.6-3.2 (1H; m), 3.2-3.8 (1H; m), 3.9-4.1 (1H; m; C _6- H), 4.1-4.9 (3H, m), 4.35 (3H; s; CH ₃ ), 4.74 (DOH), 4.9-5.3 (1H; m), 5.77 (1H; d; J = 1.2 Hz; C ₅ -H), 8.51 (1H; s; branched C ₃ -H)

HPLC (retention time): 14 minutes.

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

5% acetonitrile-water

3.65ml / min

Isomer B:

IR (KBr disc) cm ^-1 : 1765

UVλ max (H ₂ O) nm: 326

NMRδ (D ₂ O) ppm: 1.34 (3H; d; J = 6.3Hz; CH ₃ ), 2.6-3.1 (1H; m), 3.2-3.8 (1H; m), 3.9-4.1 (1H; m; C _6- H), 4.1-5.0 (3H, m), 4.36 (3H; s; CH ₃ ), 4.74 (DOH), 5.0-5.3 (1H; m), 5.81 (1H; d; J = 1.2 Hz; C ₅ -H), 8.58 (1H; s; branched C ₃ -H)

HPLC (retention time): 18 minutes.

(h) (5R, 6S, 8R) -2-[(5,6-dihydro-4H-pyrrolo [1,2-b] pyrazol-4-yl) thio] -6- (1-hydroxy Ethyl) -2-phenem-3-carboxylic acid

Isomer A

IR (KBr disc) cm ^-1 : 3380, 2950, 1760, 1580, 1365

UV lambda max (H ₂ O) nm: 252 (sh), 324

NMRδ (D ₂ O) ppm: 1.26 (3H; d; J = 5.9 Hz; CH ₃ ), 2.4-2.9 (1H; m), 2.9-3.5 (1H; m), 3.8-4.0 (1H; m; C _6- H), 4.0-4.8 (4H; m), 4.74 (DOH), 5.65 (1H; brs; C ₅ -H), 6.29 (1H; brs; branched C ₃ -H), 7.55 (1H; brs; Isomer C ₂ -H)

HPLC (retention time): 20 minutes.

Condition: same as described in (g)

Isomer B:

IR (KBr disc) cm ^-1 : 3390, 2950, 1760, 1590, 1370

UVλ max (H ₂ O) nm: 253 (sh), 324

NMRδ (D ₂ O) ppm: 1.26 (3H; d; J = 5.9 Hz; CH ₃ ), 2.4-2.9 (1H; m), 2.9-3.5 (1H; m), 3.8-4.0 (1H; m; C _6- H), 4.0-4.8 (4H; m), 5.63 (1H; brs; C ₅ -H), 6.27 (1H; brs; side chains C ₃ -H), 7.55 (1H; brs; isomers C ₂ -H )

HPLC (retention time): 24 min.

Example 15

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(5,6,7,8-tetrahydroimidazo [1,2-a] pyridin-7-yl) thio]- Isomers A and B of 2-phenem-3-carboxylic acid:

(1) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(5,6,7,8-tetrahydroimidazo [1,2-a] pyridin-7-yl) Synthesis of p-nitrobenzyl ester of thio] -2-phenem-3-carboxylic acid

Using the thiol derivative (292 mg) obtained in Reference Example 10 and repeating the same method described in Example 1- (1), the target compound (yield: 185 mg) was obtained.

NMRδ (CDCl ₃ ) ppm: 1.36 (3H; d; J = 6Hz; CH ₃ ), 1.9-2.6 (2H; m; pyridine ring C ₆ -H), 2.6-3.5 (2H; m; pyridine ring C _8- H ₂ ), 3.5-3.9 (2H, m; pyridine ring C ₇ -H and C ₆ -G), 3.9-4.3 (3H; m; pyridine ring C ₅ -H ₂ and C ₈ -H), 5.12 and 5.42 (2H; ABq; J = 14Hz; CO ₂ CH ₂ Ar), 5.70 (1H; d; J = 2Hz; C ₅ -H), 6.75 (1H; brs; imidazole ring C ₃ -H), 6.92 (1H ; brs; imidazole ring C ₂ -H), 7.53 (2H; d; J = 9 Hz; ArH), 8.12 (2H; d; J = 9 Hz; ArH)

(2) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(5,6,7,8-tetrahydroimidazo [1,2-a] pyridin-7-yl) Synthesis of Isomers A and B of Thio-2-phenem-3-carboxylic Acid

The target compounds (isomers A and B) were prepared by repeating the method of Example 1- (2) using the product (71 mg) obtained in step (1).

Isomer A: Yield 13 mg

IR (KBr disc) cm ^-1 : 1770, 1590

UVλ max (H ₂ O) nm: 260, 322

NMRδ (D ₂ O) ppm: 1.38 (3H; d; J = 6Hz; CH ₃ ), 2.1-2.6 (2H; m; pyridine ring C ₆ -H ₂ ), 3.13 (1H; dd; J = 18, 7Hz ; Pyridine ring C ₈ -H), 3.60 (1H; dd; J = 18, 5 Hz; pyridine ring C ₈ -H), 4.01 (1H; dd, J = 7, 2 Hz; C ₆ -H), 4.0-4.5 (4H; m; pyridine ring C ₅ -H ₂ pyridine ring C ₇ -H and C ₈ -H), 4.80 (HOD), 5.75 (1H; d; J = 2 Hz; C ₅ -H), 7.35 (2H; s; imidazole ring C ₂ -H and C ₃ -H)

HPLC (holding time): 22.7 minutes

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

5% acetonitrile-water

3.65ml / min

Isomer B: Yield 15 mg

IR (KBr disc) cm ^-1 : 1770, 1590

UVλ max (H ₂ O) nm: 258, 322

MMRδ (D ₂ O) ppm: 1.38 (3H; d; J = 6Hz, CH ₃ ), 2.1-2.6 (2H; m; pyridine ring C ₈ -H ₂ ), 3.17 (1H; dd; J = 18, 7Hz ; Pyridine ring C ₈ -H), 3.63 (1H; dd; J = 18, 6 Hz; pyridine ring C ₈ -H), 4.00 (1H; dd; J = 7, 2 Hz; C ₆ -H), 4.0-4.5 (4H; m; pyridine ring C ₅ -H ₂ ; pyridine ring C ₇ -H and C ₈ -H), 4.80 (HOD), 5.78 (1H; d; J = 2 Hz; C ₅ -H), 7.35 (2H s; imidazole ring C ₂ -H and C ₃ -H)

HPLC (holding time): 27.7 minutes

Example 16

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(1-methyl-5,6,7,8-tetrahydroimidazo [1,2-a] pyridinium-7- Yl) thio] -2-phenem-3-carboxylate:

Using the compound obtained in Example 15, the target compound (yield: 33 mg) was obtained according to the method of Example 2.

IR (KBr disc) cm ^-1 : 1770, 1585

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6Hz; CH ₃ ), 2.1-2.7 (2H; m; pyridine ring C ₆ -H ₂ ), 3.0-4.4 (7H; m; pyridine ring C ₅ -H _2, pyridine ring C ₇ -H, pyridine ring C ₈ -H ₂ , C ₆ -H and C ₈ -H), 3.80 (3H, s; NCH ₃ ), 4.80 (HOD), 5.80 (1H; s; C ₅ -H), 7.42 (2H; s; imidazole ring C ₂ -H and C ₃ -H)

HPLC (retention time): 14.4 min.

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

7% acetonitrile-water

3.65 ml / min.

Example 17

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(5,6,7,8-tetrahydro-1,2,4-triazolo [4,3-a] pyridine- Synthesis of 7-yl) thio] -2-phenem-3-carboxylic acid (mixture of isomers A and B):

(1) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2- [5,6,7,8-tetrahydro-1,2,4-triazolo [4,3-a] Synthesis of pyridin-7-yl) thio] -2-phenem-3-carboxylic acid:

The thiol derivative (213 mg) obtained in Reference Example 11 described later was reacted and treated according to the method of Example 1- (1) to obtain the target compound (yield: 186 mg).

NMR (CDCl ₃ ) ppm: 1.40 (3H; d; J = 6Hz; CH ₃ ), 2.0-2.6 (2H; m; pyridine ring C ₆ -H ₂ ), 3.0-4.0 (4H; m; pyridine ring C ₈ -H ₂ , pyridine ring C ₇ -H and C ₆ -H), 4.0-4.4 (3H, m; pyridine ring C ₅ -H ₂ and C ₈ -H), 5.20 and 5.48 (2H; ABq; J = 14 Hz ; CO ₂ CH ₂ Ar), 5.88 (0.5H; d; J = 2Hz; C ₅ -H), 5.91 (0.5H; d; J = 2Hz; C ₅ -H), 7.60 (2H; d; J = 9 Hz; ArH), 8.12 (1H; s; triazole ring C ₃ -H), 8.24 (2H; d; J = 9 Hz; ArH)

(2) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(5,6,7,8-tetrahydro-1,2,4-triazolo [4,3-a ] Synthesis of pyridin-7-yl) thio-2-phenem-3-carboxylic acid (mixture of isomers A and B)

The product (70 mg) obtained in the step (1) was treated according to the method of Example 1- (2) to prepare the target compound (yield: 29 mg).

IR (KBr disc) cm ^-1 : 1765, 1595

UVλ max (H ₂ O) nm: 258, 322

NMRδ (D ₂ O) ppm: 1.38 (3H; d; J = 6Hz; CH ₃ ), 2.1-2.6 (2H; m; pyridine ring C ₆ -H), 2.9-3.6 (2H; m; pyridine ring C ₈ -H ₂ ), 4.00 (1H; dd; J = 6, 2Hz; C ₆ -H), 4.0-4.5 (4H; m; pyridine ring C ₅ -H ₂ , pyridine ring C ₇ -H and C ₈ -H ), 4.80 (HOD), 5.80 (1H; d; J = 2Hz; C ₅ -H), 8.48 (1H; s; triazole ring C ₃ -H)

HPLC (retention time): 14.2 and 15.2 min.

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

3% acetonitrile-water

3.65 ml / min.

Example 18

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(1-methyl-5,6,7,8-tetrahydro-1,2,4-triazolo- [4,3 -a] Synthesis of pyridinium-7-yl) thio] -2-phenem-3-carboxylate (isomers A and B: mixture of isomers C and D)

The desired compound was prepared by repeating the method of Example 2 using the product (116 mg) obtained in Example 17.

Mixture of Isomers A and B: Yield 15 mg

IR (KBr disc) cm ^-1 : 1760, 1580

UVλmax (H ₂ O) ppm: 255, 323

NMRδ (D ₂ O) ppm: 1.36 (3H; d, J = 6Hz; CH ₃ ), 2.1-4.6 (9H; m; pyridine ring C ₆ -H ₂ , pyridine ring C ₈ -H ₂ , pyridine ring C ₅ -H ₂ , pyridine ring C ₇ -H, C ₆ -H and C ₈ -H), 4.04 (3H, s; NCH ₃ ), 4.86 (HOD), 5.80 (1H; d; J = 2Hz; C _5- H), 8.80 (1H; s; triazole ring C ₃ -H)

HPLC (retention time): 14.4 min.

Condition: Nucleosil 7C ₁₈ (10 × 300mm)

5% acetonitrile-water

3.65 ml / min.

Isomer C: Yield 7 mg

IR (KBr disc) cm ^-1 : 1765, 1590

UVλ max (H ₂ O) nm: 258, 323

NMRδ (D ₂ O) ppm: 1.39 (3H; d; J = 6Hz; CH ₃ ), 2.2-2.7 (2H; m; pyridine ring C ₆ -H ₂ ), 3.32 (1H; dd; J = 18, 7Hz ; Pyridine ring C ₈ -H), 3.68 (1H; dd; J = 18, 5 Hz; pyridine ring C ₈ -H), 4.03 (1H; dd; J = 6.2 Hz; C ₆ -H), 4.16 (3H; s; NCH ₃ ), 4.2-4.6 (4H; m; pyridine ring C ₅ -H ₂ , pyridine ring C ₇ -H and C ₈ -H), 4.80 (HOD), 5.83 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 21.2 min.

Isomer D: Yield 6 mg

IR (KBr disc) cm ^-1 : 1765, 1590

UVλ max (H ₂ O) nm: 260, 322

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6Hz; CH ₃ ), 2.3-2.6 (2H: m; pyridine ring C ₆ -H ₂ ), 3.21 (1H; dd; J = 18, 7Hz ; Pyridine ring C ₈ -H), 3.68 (1H; dd; J = 18, 5 Hz; pyridine ring C ₈ -H), 4.06 (1H; dd; J = 6, 2 Hz; C ₆ -H), 4.16 (3H s; NCH ₃ ), 4.0-4.6 (4H; m; pyridine ring C ₅ H ₂ , pyridine ring C ₇ -H and C ₈ -H), 4.80 (HOD), 5.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 29.6 min.

Reference Example A

6,7-dihydro-7-mercapto-5H-pyrrolo [1,2-c] imidazole trifluoromethane sulfonate:

(1) ethyl 3- (imidazol-4-yl) -3- (p-methoxybenzylthio) propionate:

Urucane. 3 g of ethyl ester of hydrochloride (W. Hubball and FLPyman, J. Chem. Soc., 1928, 26) are alkalized with an aqueous solution of sodium bicarbonate, saturated with NaCl, extracted with chloroform and dried over Na ₂ SO ₄ . The solution is concentrated to 50 ml under reduced pressure, 3.1 g of p-methoxysbenzyl mercaptan and 3.44 g of DBU are added and stirred under argon gas for 24 hours at room temperature. The solvent was distilled off under reduced pressure and the residue was purified by column chromatography using 40 g of silica gel and benzene-ethyl acetate (1: 1 V / V) as eluent to obtain 4.8 g of the neck compound.

NMRδ (CDCl ₃ ) ppm: 1.18 (3H; t; J = 6 Hz), 2.83-3.00 (2H; m), 3.60 (2H, s), 3.76 (3H; s), 4.07 (2H; q; J = 6 Hz ), 4.23 (1H; t; J = 6 Hz), 6.13 (1H; brs), 6.75 (2H; d; J = 9 Hz), 6.86 (1H; s), 7.13 (2H; d; J = 9 Hz), 7.53 (1H, s)

(2) 3- (imidazol-4-yl) -3- (p-methoxybenzylthio) propanol: 4.8 g of the compound obtained in step (1) was dissolved in 50 ml of THF and cooled to 0-5 캜 0.33 g of lithium borohydride is added and stirred at room temperature for 17 hours. The solvent is distilled off under reduced pressure and chloroform, water and ammonium chloride are added to the obtained residue to separate the organic phase. The organic phase is dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The desired compound was passed through a column packed with 50 g of silica gel to purify the residue and obtained after eluting the product with chloroform-methanol (95: 5 V / V) (yield: 1.98 g).

NMRδ (CDCl ₃ ) ppm: 1.90-2.30 (3H; m), 3.50 (2H; s), 3.69 (2H, s), 3.94 (1H; t; J = 6Hz), 6.69 (2H; d; J = 9Hz ), 6.81 (1H; s), 7.06 (2H; d; J = 9 Hz), 7.49 (1H, s)

(3) 3- (imidazol-4-yl) -3- (p-methoxybenzylthio) propylchloride

1.2 g of the compound obtained in step (2) is dissolved in a mixed solvent of THF (30 ml) and carbon tetrachloride (30 ml), and 2.26 g of triphenylphosphine are added and stirred at 50 ° C. for 17 hours. The reaction mixture is distilled off under reduced pressure and the resulting residue is purified by column chromatography using 30 g of silica gel and benzene-methanol (97: 3 V / V) as eluent. This yields the target compound in a yield of 0.41 g.

NMRδ (CDCl ₃ ) ppm: 2.15-2.50 (2H; m), 3.55 (2H; s), 3.74 (3H; s), 4.00 (1H; t; J = 6Hz), 6.74 (2H; d; J = 9Hz ), 6.86 (1H; s), 7.12 (2H; d; J = 9 Hz), 7.57 (1H; s)

(4) 6.7-dihydro-7- (p-methoxybenzylthio) -5H-pyrrolo [1,2-c] imidazole:

400 mg of the compound obtained in the above step (3) is dissolved in 10 ml of acetone, 2.0 g of sodium iodide is added and stirred at room temperature for 17 hours. The mixture is heated to reflux with heating for 8 hours and the solvent is distilled off under reduced pressure. To the residue is added chloroform, water and sodium carbonate (3 g), the organic phase is separated, dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The residue was purified by column chromatography using 10 g of silica gel and chloroform-methanol (99: 1 V / V) as eluent to obtain the target compound (yield: 250 mg).

NMRδ (CDCl ₃ ) ppm: 2.27-2.60 (1H; m), 2.65-3.15 (1H; m), 3.70 (2H; s), 3.75 (3H; s), 3.80-4.20 (3H; M), 6.76 ( 1 H; s), 6.78 (2 H; d; J = 9 Hz), 7.34 (1 H; s)

(5) 6,7-dihydro-7-mercapto-5H-pyrrolo [1,2-c] imidazole trifluoromethane sulfonate:

6,7-dihydro-7-p-methoxy benzylthio-5H-pyrrolo [1,2-c] -imidazole (400 mg) obtained in step (4), anisole (1.5 ml), Trifluoro-acetic acid (9 ml) and trifluoromethanesulfonic acid (0.36 ml) are added and stirred at room temperature for 30 minutes. The reaction solution is concentrated under reduced pressure and further concentrated after addition of xylene (200 ml). This method is repeated twice and the resulting residue is washed with ether. The ether was decanted and dried under reduced pressure to yield the target compound (yield: 440 mg).

NMRδ (CDCl ₃ ) ppm: 2.50-2.80 (1H; m), 3.00-3.43 (1H; m), 4.20-4.70 (3H: m), 4.80 (HOD), 7.33 (1H; 2; imidazole-H) 8.60 (1H; s; imidazole-H)

Reference Example 1

Synthesis of 6,7-dihydro-7-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoro-methane sulfonate:

The method of Reference Example A was followed using 6,7-dihydro-7-p-methoxybenzylthio-5H-pyrrolo [2,1-c] -1,2,4-triazole (280 g). Repeatedly obtained the target compound (yield: 361 mg colorless transparent oil).

NMRδ (DMSO-d ₆ + D ₂ O) ppm: 1.8-2.6 (1H; m), 3.0-3.6 (1H; m), 4.1-4.5 (2H; m; C ₅ -H), 4.65 (1H; dd ; J = 8), 9.50 (1H; s)

Reference Example 2

6,7-dihydro-6-mercapto-3-trifluoromethyl-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethane sulfonate

According to the same method as described in Reference Example A, the above-mentioned compound was added to 6,7-dihydro-6-p-methoxybenzylthio-3-trifluoromethyl-5H-pyrrolo [2,1-c. ] -1,2,4-triazole (116 mg) as a pale yellow oil (231 mg).

NMRδ (D ₂ O) ppm: 3.08 (1H; dd; J = 18, 7 Hz; C ₇ -H), 3.68 (1H; dd; J = 18, 7 Hz, C ₇ -H), 4.1-4.7 (3H; m; C ₅ -H ₂ and C ₆ -H), 4.80 (HOD)

The following compounds are also prepared following the same method as described in Reference Example A.

Reference Example 3

3-amino-6,7-dihydro-7-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethane sulfonate

Reference Example 4

3-chloro-6,7-dihydro-7-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 2.50-2.95 and 3.15-3.60 (1H; m; C ₆ -H), 4.05-4.80 (3H; m, C ₅ -H ₂ , C ₇ -H), 4.80 (HOD)

Reference Example 5

2-amino-6,7-dihydro-7-mercapto-5H-pyrrolo [2,1-b] -1,2,4-triazole trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 2.60-2.90 and 3.18-3.60 (1H; m), 4.10-4.75 (3H; m), 4.80 (HOD)

Reference Example 6

2-chloro-6,7-dihydro-7-mercapto-5H-pyrrolo [2,1-b] -1,2,4-triazole trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 2.49-2.80 and 3.10-3.50 (1H; m), 4.05-4.58 (3H; m), 4.80 (HOD)

Reference Example 7

6,7-dichloro-6-mercapto-5H-pyrrole [1,2-c] thiazolium trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 3.15 (1H; dd), 3.70 (1H; dd), 4.2-5.2 (3H; m), 4.74 (DOH), 7.84 (1H; d; J = 1.9 Hz), 9.83 ( 1H; d; J = 19)

Reference Example 8

5,6-dihydro-4-mercapto-4H-pyrrolo [1,2-c] -1,2,3-triazole trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 2.5-3.1 (1H; m), 3.2-3.7 (1H; m), 4.4-5.1 (3H; m), 4.74 (DOH), 8.20 (1H; s)

Reference Example 9

5,6, -dihydro-4-mercapto-4H-pyrrole [1,2-b] pyrazole trifluoromethane sulfonate

NMRδ (D ₂ O) ppm: 2.4-3.0 (1H; m), 3.0-3.6 (1H; m), 4.1-4.8 (3H; m), 4.74 (DOH), 6.55 (1H; d; J = 3 Hz) , 7.98 (1H; d; J = 3 Hz)

Reference Example 10

7-mercapto-5,6,7,8-tetrahydroimidazo [1,2-a] pyridine trifluoromethane sulfonate

Using 7-p-methoxybenzylthio-5,6,7,8-tetrahydroimidazo [1,2-a] pyridine (249 mg), the target compound was prepared in the same manner as described in Reference Example A. To produce accordingly. Its yield is 292 mg (colorless transparent oil).

Reference Example 11

7-mercapto-5,6,7,8-tetrahydro-1,2,4-triazolo [4,3-a] pyridine trifluoromethane sulfonate

7-p-methoxybenzylthio-5,6,7,8-tetrahydro-1,2,4-triazolo [4-3-a] pyridine starting from 307 mg, the same as described in Reference Example A The process is repeated to give 435 mg of the target compound as the product of a clear colorless oil.

NMRδ (D ₂ O + DMSO-d ₆ ) ppm: 2.1-2.7 (2H; m), 3.31 (1H; dd; J = 18,7Hz), 3.80 (1H; dd; J = 18,7Hz), 3.6- 3.9 (1H; m; C ₇ -H), 4.4-4.6 (2H; m; C ₅ -H ₂ ), 9.19 (1H; s; C ₃ -H)

Example 19

(5R, 6S, 8R) -2-[(2,3, -dihydro-1H-indolizinium-1-yl) thio] -6- (1-hydroxyethyl) -phenem-3-carboxylate synthesis

180 mg of p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -phenem-3-carboxylate in 3 ml of DMF (Japanese Patent Laid-Open No. 56987/1985 Prepared according to the method of Example 1) and 271 mg of 2,3-dihydro-1-mercapto-1H-indoleginium trifluoromethane sulfonate and then cooled under -40 DEG C under nitrogen atmosphere. Add 108 mg of isopropylethyl amine and stir at −40 ° C. for 30 minutes. The reaction solution is added to ether and centrifuged to remove the supernatant. The precipitate is dissolved in 40 ml of 50% THF-water, to which 4.16 g of ammonium chloride and 2.08 g of iron powder are added under ice cooling, followed by vigorous stirring at this temperature for 50 minutes. The reaction solution was filtered through Celite (Celite: Johns-Manville) and the filtrate was concentrated under reduced pressure and washed with ethyl acetate. The aqueous phase is concentrated in vacuo and the concentrate is purified through a Diaion column (Diaion HP-20, 1.8 cm × 22 cm). The fraction eluted with 350 ml of water was discarded and the fraction eluted with 5% THF-water was concentrated in vacuo and then purified by HPLC (carrier: Nucleosil 7C ₁₈ 10 mm × 30 mm; solvent: 7% acetonitrile-water; flow rate: 3.65 ml / min) Purification and lyophilization gave the desired compound as pale yellow powder (yield: 30 mg).

IR (KBr disc) cm ^-1 : 1760, 1590

UVλ max (H ₂ O) nm: 265, 325

NMRδ (D ₂ O) ppm: 1.33 (3H; d; J = 6Hz, CH ₃ ), 2.48-2.92 (1H; m, indolinium ring C ₂ -H), 2.92-3.40 (1H; m; indolinium ring C ₂ -H), 3.95-5.20 (5H, m; indolizium ring C ₁ -H and C ₃ -H ₂ , C ₆ -H and C ₈ -H), 4.80 (HOD), 5.64 (1 / 3H; d; J = 2Hz; C ₅ -H), 5.76 (2 / 3H; D; J = 2Hz; C ₅ -H), 7.92-8.32 (2H; m; indolinium ring C ₆ -H and C ₈ -H ), 8.58 (1H; dt; J = 1, 7 Hz; indolinium ring C ₇ -H), 8.88 (1H; dd; J = 1, 7 Hz; indolinium ring C ₅ -H), HPLC (holding time): 13.0 minutes and 13.6 minutes.

Elemental analysis _{(%): (C 10 H} 16 N 2 O 4 S 2 · H 2 O)

Theoretic value: C; 49.98, H; 4.72, N; 7.29

Found: C; 50.22, H; 4.75, N; 7.38

Example 20

Preparation of (5R, 6S, 8R) -2-[(2,3-dihydro-1H-indolizinium-2-yl) thio] -6- (1-hydroxyethyl) -phenem-3-carboxylate

According to a method analogous to Example 19, 2,3-dihydro-2-mercapto-1H-indolizinium trifluoromethane sulfonate is used to obtain the pale yellow target compound.

Isomer A:

UV λ max (H ₂ O) nm: 264, 322

HPLC (retention time): 11.7 minutes

Carrier: Nucleosil 7C ₁₈ (10 × 300mm)

Solvent: 7% Acetonitrile-water

Flow rate: 3.65 ml / min

Isomer B:

UV λ max (H ₂ O) nm: 264, 322

HPLC (holding time): 13.4 minutes

Example 21

(5R, 6S, 8R) -2-[(6,7, -dihydro-5H-pyrrolo- [1,2-a] imidazol-7-yl) thio] -6- (1-hydroxyethyl Preparation of Phenem-3-carboxylic Acid

(1) p-nitrobenzyl (5R, 6S, 6R) -6- (1-hydroxylethyl) -2-dihydro-5H-pyrrolo [1,2-a] imidazol-7-yl) thio] -Phenem-3-carboxylate Synthesis

p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -phenem-3-carboxylate (190 mg) and 6,7-dihydro-7-mercapto -5H-pyrrolo [1,2, -a] imidazole trifluoro methanesulfonate (261 mg) was dissolved in 5 ml of DMF and 232 mg of diisopropylethylamine was added thereto while cooling at -50 ° C. After stirring at the same temperature for 30 minutes. The reaction solution is diluted with ethyl acetate, washed, dried over MgSO ₄ and distilled under reduced pressure. The residue was purified by column chromatography (using 18 g of silica gel, chloroform: methanol = 5: 1 V / V) to obtain a caramel target compound (yield: 178 mg).

NMRδ (CDCl ₃ ) ppm: 1.40 (3H; d; J = 6Hz; CH ₃ ), 2.6-3.5 (2H; m; pyrroline ring C ₆ -H ₂ ), 3.7-4.4 (4H; m, pyrroline ring C ₅ -H ₂ , C ₆ -H and C ₈ -H), 5.18 and 5.36 (0.5H each; ABq; J = 14 Hz; CO ₂ CH ₂ Ar), 5.28 and 5.41 (0.5H each; ABq; J = 14Hz) ; CO ₂ CH ₂ Ar), 5.75 (0.5H; d; J = 2Hz; C ₅ -H), 5.98 (0.5H; d; J = 2Hz; C ₅ -H), 6.92 (0.5H, d; J = 2 Hz; imidazole ring-H), 6.96 (0.5H; J = 2 Hz; imidazole ring-1H), 7.58 (1H; d; 9 Hz, ArH), 7.64 (1H; D; J = 9 Hz; ArH), 8.16 (1H; d; J = 9 Hz; ArH), 8.24 (1H; d; J = 9 Hz; ArH).

(2) (5R, 6S, 8R) -2-[(6,7-dihydro-5H-pyrrolo [1,2-a] imidazol-7-yl) thio] -6- (1-hydroxy Synthesis of ethyl) phenem-3-carboxylic acid

The compound (178 mg) obtained in the above step (1) was dissolved in a mixture of THF (15 ml) and phosphate buffer (15 ml; pH7.0), followed by addition of 200 mg of 10% palladium-carbon catalyst and reduction at room temperature for 2 hours. React. The catalyst is filtered off, the filtrate and washings are combined, concentrated and washed with ethyl acetate. The water phase is concentrated and purified by column chromatography (Diaion HP-20, 1.8 cm × 20 cm).

The fraction eluted with 50 ml of water was discarded, and the fraction eluted with 150 ml of water and the fraction eluted with 80 ml of 5% THF-water were concentrated and concentrated by HPLC [carrier; Nucleosil 7C ₁₈ (10 × 30 mm); Solvent: 5% acetonitrile-water; Flow rate: 3.65 ml / min] treatment and lyophilization yielded 58 mg of the target compound as a colorless powder.

IR (KBr disc) cm ^-1 : 1760, 1580

UV λ max (H ₂ O) nm: 325

NMRδ (D ₂ O) ppm: 1.33 (1.5H; d; J = 6Hz; CH ₃ ), 1.36 (1.5H; d; J = 6Hz; CH ₃ ), 2.8-3.1 (1H; m; pyrroline ring C ₆ -H), 3.1-3.5 (1H; m; pyrroline ring C ₆ -H), 4.0-4.6 (4H; m; pyrroline ring C ₅ -H ₂ , C ₆ -H and C ₈ -H), 4.80 (HOD ), 5.66 (0.5H; d; J = 2 Hz; C ₅ -H), 5.73 (0.5H; d; J = 2 Hz, C ₅ -H), 7.2-7.4 (2H; m; imidazole ring-H) HPLC (retention time): 13.6 min.

Example 22

p-nitrobenzyl (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [1,2-a] imidazol-6-yl Synthesis of Thio] -2-phenem-3-carboxylate

p-nitrobenzyl (5R, 6S, 8R) -2-ethyl sulfinyl-6- (1-hydroxyethyl) -phenem-carboxylate (213 mg) was added to a mixture of acetonitrile (5 ml) and THF (2 ml). Dissolve and cool to -40 ° C to -50 ° C. To the solution, 6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-a] imidazole trifluoromethane sulfonate (390 mg) was dissolved in THF (3 ml) and DBU (410 mg). Is added to the solution dissolved in and stirred at this temperature for 20 minutes. After the reaction was completed, 30 ml of ethyl acetate was added, washed with water and saturated NaCl solution, dried over Na ₂ SO ₄ , and concentrated under reduced pressure. The residue is passed through a column (15 g of silica gel) and eluted with chloroformmethanol (19: 1 V / V). The eluate containing the target compound is collected and concentrated. 225 g of the desired compound in the yellow oil phase are obtained.

), 3.7-4.7(5H ; m ;

NMRδ (CDCl ₃ ) ppm: 1.38 (3H; d; J = 6 Hz; CH ₃ ), 2.8-3.7 (2H; m;

), 3.7-4.7 (5H; m;

, C ₆ -H, C ₈ -H), 5.32 (2H; ABq; J = 14 Hz; -COO-CH _2- ), 5.76 (1H; d; J = 2 Hz; C ₅ -H), 6.87 and 7.05 ( Each 1H; each d; J = 1 Hz; imidazole ring-H), 7.59 and 8.18 (each 2H; each d; J = 9 Hz; -C ₆ H ₄ NO ₂ ) TLC (thin layer chromatography), Rf = 0.4

Condition: silica gel plate (Merck: 0.25mm thickness)

Developing Solvent: Chloroform-Methanol (9: 1V / V)

(2) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [1,2-a] imidazol-6-yl) -Thio] -2-phenem-3-carboxylic acid (isomers A and B)

The compound (84 mg) obtained in step (1) was dissolved in a mixture of THF (5 ml) and phosphate buffer (5 ml; pH 7), and then 10% palladium-carbon catalyst (160 mg) was added thereto, followed by hydrogen atmosphere at 1 atm. Under reduced temperature at room temperature for 3 hours. After the catalyst is filtered off, the filtrate is diluted with water and washed with an ether-ethyl acetate mixed solvent (1: 1 V / V). The water phase is concentrated to about 30 ml under reduced pressure and purified by passing through a column (Diaion HP-20, 1.8 cm x 22 cm). Fractions eluted with 100 ml water are discarded and concentrated by collecting fractions eluted with 5% THF-water (showing maximum UV absorbance near 320 nm). The concentrate was purified by HPLC [carrier: nucleosil 7C ₁₈ , 10 × 30 mm; Solvent; 5% acetonitrile-water; Flow rate; 4 ml / min], fractions containing the desired compound and having a retention time of 14 minutes were concentrated under reduced pressure, and then lyophilized to give the desired product as a pale yellow powder (yield: 24 mg).

IR (KBr disc) cm ^-1 : 3400, 1765, 1585.

UV λ max (H ₂ O) nm: 323

NMR δ (D ₂ O) ppm: 1.37 (3H, d, J = 6 Hz, CH ₃ ), 3.2 and 3.6 (each 1H, each m,

, 4.02 (1H, doublet of doublets, J = 2, 6 Hz; C ₆ -H), 4.1-4.5 (4H, m,

C ₈ -H), 4.80 (HOD), 5.79 (1H, d, J = 2 Hz, C ₅ -H), 7.22 (2H, brs, imidazole ring-H).

The product was purified by HPLC [carrier: C ₁₈ -octadecylsilane (manufactured by SSC-ODS-762 Senso Furniture Co., Ltd.), 20 × 300 mm; Solvent: 6% acetonitrile-water; Flow rate: 10 ml / min] Collect the fractions containing the target compound and holding time of 15 min and 16.5 min, respectively. Thus, lyophilization treatment gives Isomers A and B as pale yellow powder.

Isomer A:

NMRδ (D ₂ O) ppm: 1.37 (3H, d, J = 6 Hz, CH ₃ ), 2.96-3.23 (1H, m), 3.46-3.80 (1H, m), 4.00 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.10-4.50 (2H, m), 4.50-4.75 (2H, m), 4.80 (HOD), 5.80 (1H, d, J = 2 Hz, C ₅ -H), 7.30 (2H , s, imidazole ring-H), HPLC (holding time): 15 minutes.

Isomer B:

NMRδ (D ₂ O) ppm: 1.36 (3H, d, J = 6 Hz, CH ₃ ), 3.14-3.44 (1H, m), 3.63-3.93 (1H, m), 3.99 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.10-4.50 (2H, m), 4.80 (HOD), 5.78 (1H, d, J = 2 Hz, C ₅ -H), 7.35 (2H, s, imidazole ring-H) , HPLC (holding time): 16.5 minutes

Example 23

(5R, 6S, 8R) -6- (1-hydroxymethyl) -2-[(6,7-dihydro-1-methyl-5H-pyrrolo [1,2-a] imidazolium-6- Yl) thio] -2-phenem-3-carboxylate synthesis

The compound (220 mg) obtained in step (1) of Example 22 was dispersed in a mixture of acetone (3 ml) and THF (5 ml), and 0.84 ml of methyl iodide was added under ice cooling, followed by reaction at 3 ° C. for 16 hours. The reaction solution is concentrated under reduced pressure and the residue is washed with ether and dried. The powder obtained here was dissolved in a mixed solution of 20 ml of THF and 20 ml of water, 4.4 g of ammonium chloride and 2.2 g of iron powder (10 mesh) were added, followed by vigorous stirring for 1 hour. Remove insolubles using Celite (Johns-ManVille), dilute the filtrate with water, wash with ethyl acetate-ether (1: 1) mixture, concentrate the water phase under pressure to 30ml. Was purified by passing through a column (1.8 cm x 2.3 cm) filled with Diaion HP-20. The fraction eluted with 100 ml of water was removed and the fraction eluted with 5% THF-water was collected and concentrated under reduced pressure. Treatment of this concentrate by HPLC [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent 3% acetonitrile-water; Flow rate: 4 ml / min], containing the target compound, the fraction having a retention time of 23 minutes was collected, concentrated in vacuo, and dried to obtain the target compound (yield: 54 mg; grayish yellow powder).

IR (KBr disc) cm ^-1 : 3400, 1770, 1590

UV λ max (H ₂ O nm: 324

), 3.83 (3H; s; N-CH ₃ ), 4.01 (1H, dd, J = 2, 6 Hz, C ₆ -H), 4.2-4.5 (4H, m, S-

-CH₂-N, C₈-N), 4.80(HOD), 5.79 (1H, d, J=2Hz, C₅-H), 7.43(2H, s ; 미다졸륨환-H).

-CH ₂ -N, C ₈ -N), 4.80 (HOD), 5.79 (1H, d, J = 2 Hz, C ₅ -H), 7.43 (2H, s; midazolium ring-H).

Example 24

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [1,2-c] imidazol-6-yl) -thio] 2-phenem-3-carboxylic acid (isomers A and B) synthesis

(1) p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [1,2 -c] imidazol-6-yl) thio] -2-phenem-3-carboxylate synthesis

p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -2-phenem-3-carboxylate (213 mg) in THF (4 ml) and acetonitrile (3 ml ), And stirred at -50 ° C. 6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-c] -imidazole trifluoromethanesulfonate (360 mg) obtained in Reference Example (6-16) was THF (3 ml). ) And DBU 380 mg were added to the above solution and stirred at -50 ° C for 15 minutes. To this reaction solution was added ethyl acetate (30 ml), washed with water, washed with saturated NaCl solution, dried over Na ₂ SO ₅ and concentrated under reduced pressure. The residue obtained here was subjected to column chromatography using silica gel [eluent = CHCl ₃ ; CH ₃ OH (19: 1 V / V)] affords the target compound as a yellow oil (yield: 94 mg). (Some of them solidified)

), 7.45(1H;s;

), 7.60(2H;d;J=9Hz; 2×ArH), 8.20(2H;d;J=9Hz, 2×ArH).NMRδ (CDCl ₃ ) ppm: 1.39 (3H; d; J = 6 Hz, CH ₃ ), 2.83-3.6 (2H; m), 3.81 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.0- 4.6 (4H; m), 5.33 (2H; ABq; J = 14 Hz; -OCH ₂ Ar), 5.76 (1H; d; J = 2 Hz; C ₅ -H), 6.76 (1H; s;

), 7.45 (1 H; s;

), 7.60 (2H; d; J = 9 Hz; 2 x ArH), 8.20 (2H; d; J = 9 Hz, 2 x ArH).

(2) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [1,2-c] imidazol-6-yl) Thio] -2-phenem-3-carboxylic acid (isomers A and B) synthesis

The compound (94 mg) obtained in the previous step (1) was dissolved in a mixture of THF (7 ml) and phosphate buffer (7 ml; pH7) and 4.5 mg at room temperature under 1 atm hydrogen pressure with 180 mg of 10% Pd-C. Contact reduction for time. The catalyst is filtered off and the filtrate is diluted with water and washed with a 1: 1 mixture of ether and ethyl-acetate. The aqueous phase is concentrated to 30 ml and purified by passing through a column (1.8 cm × 23 cm) filled with Diion HP-20. The fraction eluted with 100 ml of water was removed, and the fractions eluted with 5% acetonitrile-water were collected and concentrated under reduced pressure at 323 nm. This concentrate was treated by HPLC [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent: 5% acetonitrile-water; Flow rate: 4 ml / min], the fractions with residence times 16 minutes and 21 minutes were collected, concentrated in vacuo, and lyophilized to obtain the desired compound isomers A and B of an off-yellow powder.

Isomer A: Yield 13 mg

IR (KBr disc) cm ^-1 : 3400, 1765, 1620, 1580

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz; CH ₃ ), 3.0-3.8 (2H; m), 4.01 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.20 -4.50 (4H; m), 4.80 (HOD), 5.79 (1H; d; J = 2Hz; C ₅ -H), 7.11 (1H; s; imidazole ring-H), 8.37 (1H; s; imida Sol ring-H)

HPLC (retention time): 16 minutes.

Isomer B: Yield 10 mg

IR (KBr disc) cm ^-1 : 3400, 1765, 1585

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d, J = 6 Hz; CH ₃ ), 2.9-3.80 (2H; m), 4.01 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.20 -4.60 (4H; m), 4.80 (HOD), 5.78 (1H; d; J = 2 Hz, C ₅ -H)., 7.11 (1H; s; imidazole ring-H), 8.36 (1H; s; already Dazole ring-H)

HPLC (retention time): 21 minutes.

Example 25

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-2-methyl-5H-pyrrolo [1,2-c] imidazolium-6- Yl) thio] -2-phenem-3-carboxylate (isomers A and B)

The compound (60 mg) obtained in step (1) of Example 24 was dissolved in 4 ml of acetone, 0.5 ml of methyl iodide was added, and then left at 3 ° C. for 3 days. The reaction solution was evaporated under reduced pressure, and the powder obtained was dissolved in a mixture of THF (10 ml) and water (10 ml), 1.2 g of ammonium chloride and 600 mg of iron powder (100 mesh) were added, followed by vigorous stirring for 1 hour. The insolubles are filtered using celite and the filtrate is diluted with water, washed with a 1: 1 mixture of ethyl acetate-ether and the water phase is concentrated under reduced pressure to 20 ml. The concentrate was economiced by column chromatography using Diion HP-20 (1.8 cm × 20 cm), discarded the fraction eluted with 100 ml of water, and the fraction eluted with 5% THF-water was collected and concentrated under reduced pressure. The concentrate obtained here was subjected to HPLC treatment [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent; 5% acetonitrile-water; Flow rate; 4 ml / min] and a retention time of 18 minutes and 24 minutes, respectively, are obtained in the form of an off-yellow powder, which are the present compound isomers A and B.

Isomer A: Yield 9 mg

IR (KBr disc) cm ^-1 : 1760, 1725, 1670, 1640, 1580

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz; CH ₃ ), 3.22 (1H; dd; J = 18, 4 Hz), 3.68 (1H; dd; J = 18, 7 Hz), 3.94 ( 3H; s; ≡N ⁺ CH ₃ ), 4.01 (1H; dd; J = 7, 2 Hz, C ₆ -H), 4.20-4.70 (4H; m), 4.80 (HOD), 5.78 (1H; d; J = 2 Hz; C ₅ -H), 7.23 (1H; s; imidazolium ring-H), 8.64 (1H; s; imidazolium ring -H)

HPLC (retention time): 18 minutes.

Isomer B: Yield 6 mg

IR (KBr disc) cm ^-1 : 1765, 1590

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz, CH ₃ ), 3.12 (1H; dd; J = 18, 4 Hz), 3.64 (1H; dd; J = 18, 7 Hz), 3.94 ( 3H; s; ≡N ⁺ CH ₃ ), 4.20-4.70 (4H; m), 4.80 (HOD), 5.78 (1H; d; J = 2Hz; C ₅ -H), 7.24 (1H; s; imidazolium ring -H), 8.64 (1H, s, imidazolium ring-H9.

HPLC (retention time): 24 min.

Example 26

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-1-hydroxy-5H-pyrrolo [1,2-c] imidazole-6- Yl) -thio] -phenem-3-carboxylic acid synthesis

(1) p-nitrobenzyl (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-1-hydroxy-5H-pyrrolo [1,2- c] -imidazol-6-yl) thio] -2-phenem-3-carboxylate synthesis

P-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfonyl-6- (1-hydroxyethyl) -2-phenem-3-carboxylate 170 according to a method analogous to step (1) of example 22 Mg, 300 mg of 6,7-dihydro-1-hydroxy-6-mercapto-5H-pyrrolo [1,2-c] imidazole obtained in step (8) of Reference Example 17 and 396 mg of DBU To prepare the target compound.

Yield: 195 mg.

NMRδ (CDCl ₃ -CD ₃ OD) ppm: 1.35 (3H; d; J = 6 Hz, CH ₃ ), 5.35 (2H; ABq; J = 14 Hz; OCH ₂ Ar), 5.72 (1H; d; J = 2 Hz, C ₅ -H), 7.64 (2H; d; J = 9 Hz, 2 x ArH), 8.23 (2H; d; J = 9 Hz; 2 x ArH), 8.23 (1H; s; imidazole ring-H).

(2) (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-1-hydroxy-5H-pyrrolo [1,2-c] imidazole -6-yl) thio] -2-phenem-3-carboxylic acid synthesis

According to a method similar to that in step (2) of Example 22, 195 mg of the compound obtained in the preceding step (1) and 300 mg of 10% Pd-C were used to obtain the target compound (yield: 36). Mg).

IR (KBr disc) cm ^-1 : 3400, 1760, 1650, 1580.

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz, CH ₃ ), 2.60 (2H; m), 4.00-4.40 (5H; m), 4.80 (HOD), 5.78 (1H; d; J = 2 Hz, C ₅ -H), 8.32 (1H; s; imidazole ring-H) (holding time): 13 minutes.

Condition: Carrier: Nucleosil 7C ₁₈ (10 × 300mm)

Solvent: 1% acetonitrile-water

Flow rate: 4ml / min

Example 27

(1) p-nitrobenzyl (5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [2,1, -c] -1 , 2,4-triazol-6-yl) thio-2-phenem-3-carboxylate synthesis

p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6-1-hydroxyethyl) phenem-3-carboxylate (170 mg) in a mixture of acetonitrile (2 ml) and THF (2 ml) Dissolve and cool to -30 ° C. To this cooled solution 6,7-dihydro-6-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole-trifluoromethane sulfonate (250 mg) and aceto A solution of nitrile (2 ml) and DBU (244 mg) is added and stirred at -30 ° C for 20 minutes. After the reaction was completed, 30 ml of ethyl acetate was added, washed with water, washed with saturated aqueous NaCl solution, dried over Na ₂ SO ₄ , and concentrated under reduced pressure. The residue is treated by column chromatography using silica gel and eluted with methanol containing chloroform (gradually increasing the polarity of the eluate by changing the mixing ratio between CHCl ₃ and CH ₃ OH from 19: 1 to 4: 1). Fractions containing the target compound were collected and concentrated under reduced pressure to yield the target compound in a yellow oil state (yield: 147 mg).

NMRδ (CDCl ₃ + CD ₃ OD) ppm: 1.36 (3H; d; J = 6 Hz, CH ₃ ), 3.0-3.9 (3H; m), 4.0-4.8 (4H; m), 5.34 (2H; ABq, J ^{_{= 14Hz; -COOCH 2 -),}} 5.77 (1H; d; J = 2Hz; C 5 H), 7.61 and 8.21 (each 2H; each _{d; J = 9Hz; C 6} H 4 NO 2), 8.18 (1H; s; triazole ring-H).

TLC: Rf = 0.3 (silica gel plate (Merck), thickness: 0.25 developing solvent: chloroform-methanol (4: 1 V / V)]

(2) (5R, 6S, 8S) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-5H-pyrrolo [2,1-c] 1,2,4-tria Zol-6-yl) thio] -2-phenem-3-carboxylic acid (isomers A and B) synthesis

The compound (141 mg) obtained in the above step (1) was dissolved in a mixture of THF (6 ml) and phosphate buffer (pH 7.0; 6 ml), 200 mg of 10% Pd-C was added and 3.5 at 1 atmosphere of hydrogen at room temperature. Contact reduction for time. After filtration of the catalyst the filtrate is diluted with water and the water phase is washed with a 1: 1 mixture of ether and ethyl acetate. The water phase is concentrated to 30 ml, which is passed through a column (1.8 × 20 cm) filled with Diion HP-20 and purified. The fraction eluted with 100 ml of water was discarded and the fraction eluted with 5% THf-water was collected and concentrated under reduced pressure. The concentrate was subjected to HPLC again [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent: 3% acetonitrile-water; Flow rate; 4 ml / min], the fraction having a retention time of 11 minutes was collected, concentrated under reduced pressure, and lyophilized to yield the target compound as a colorless powder (30 mg).

IR (KBr disc) cm ^-1 : 3400, 1760, 1580

UVλ max (H ₂ O) nm: 323

), 4.00 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.2-4.4 (m), 4.80 (HOD), 5.77 (1H; d; J = 2 Hz; C ₅ -H), 8.44 ( 1H; s; triazole ring-H), this product was subjected to HPLC again [carrier: ODS (20 x 300 mm); Solvent; 3% acetonitrile-water; Flow rate: 10 ml / min] and fractions having retention times of 12.3 and 13 minutes are collected and lyophilized to obtain Isomers A and B as off-yellow powders.

Isomer A:

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz; CH ₃ ), 3.00-3.32 (1H; m), 3.50-3.83 (1H; m), 4.00 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.10-4.50 (2H; m), 4.80 (HOD), 5.77 (1H; d; J = 2 Hz; C ₅ -H), 8.44 (1H; s; triazole ring-H)

Isomer B:

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz, CH ₃ ), 2.96-3.22 (1H; m), 3.46-3.78 (1H; m), 4.00 (1H; dd; J = 2, 6 Hz; C ₆ -H), 4.13-4.50 (2H; m), 4.80 (HOD), 5.77 (1H; d; J = 2 Hz; C ₆ -H), 8.44 (1H; s; triazole ring-H)

Example 28

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(6,7-dihydro-N-methyl-5H-pyrrolo [2,1-c] -1,2,4 -Triazol-6-yl) thio] -2-phenem-3-carboxylate (mixture of isomers A and B, mixture of isomers C and D)

The compound (400 mg) obtained in the step (1) of Example 27 was treated in a similar manner to the method described in Example 23, followed by HPLC treatment [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent: 5% acetonitrile-water; Flow rate 5 ml / min] to separate and purify the product. Thus, three fractions having retention times of 7.8 minutes, 8.4 minutes, 9.7 minutes and 13.5 minutes, respectively, were separated and lyophilized to obtain the desired compound as a yellow powder in which isomers A, B and isomers C and D were mixed.

Mixed isomers A and B: Yield 35 mg

IR (KBr disc) cm ^-1 : 3400, 1770, 1640, 1580

UVλ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz, CH ₃ ), 3.3-4.0 (2H; m), 4.06 (3H; s; N-CH ₃ ), 4.80 (HOD), 5.80 ( 1H; d; J = 2 Hz, C ₅ -H), 8.76 (1H; s; triazolium ring-H)

Isomer C: Yield 36 mg

UV λ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.38 (3H; d; J = 6 Hz), 3.25-3.56 (1H; m), 3.71-4.10 (2H; m), 4.18 (3H; s; N-CH ₃ ), 4.80 (HOD), 5.81 (1H; d; J = 2Hx, C ₅ -H)

Isomer D: Yield 38 mg

UV λ max (H ₂ O) nm: 323

NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6Hz, CH ₃ ), 3.15-3.45 (1H; m), 3.63-4.10 (2H; m), 4.18 (3H; s; N-CH ₃ ), 4.80 (HOD), 5.81 (1H; d; J = 2 Hz; C ₅ -H)

Example 29

(5R, 6S, 8R) -2-[(3-amino-6,7-dihydro-5H-pyrrolo [2,1-c] -1,2,4-triazol-6-yl) thio] Synthesis of -6- (1-hydroxyethyl) phenem-3-carboxylic acid

p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -phenem-3-carboxylate (70 mg) and 3-amino-6,7-dihydro- 6-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethane sulfonate (100 mg) was dissolved in DMF (2 ml) and cooled to -50 ° C. Isopropyl ethylamine (85 mg) was added and stirred at -50 ° C for 30 minutes. The reaction solution is poured into ether and the supernatant is decanted off. The precipitate is dissolved in a mixture of THF (15 ml) and phosphate buffer (pH 7.0; 15 ml) and 10% Pd-C (100 mg) is added for catalytic reduction at room temperature for 5 hours. The catalyst is filtered off and the filtrate is concentrated under reduced pressure and washed with ethyl acetate. The water phase is concentrated and this concentrate is purified by passing through a column (8 × 18 cm) filled with Diion HP-20. In this step, the fraction eluted with 130 ml of water was discarded, and the fraction eluted with 5% THF-water was collected, collected, concentrated and treated by HPLC [carrier: nucleosil 7C ₁₈ (10 × 300 mm); Solvent: 3% acetonitrile-water: flow rate; 3.65 ml / min. Lyophilization yielded 24 mg of the target compound as a colorless powder.

IR (kBr disc) cm ⁻¹ ; 1760, 1675

UVλmax (H ² O): 253, 323

NMRδ (D ₂ O) ppm: 1.36 (9 / 5H; d; J = 6Hz; CH ₃ ), 1.40 (6 / 5H; d; J = 6Hz; CH ₃ ), 3.1-3.5 (1H; m; pyrroline ring C ₇ -H), 3.50-4.8 (1H; pyrroline ring C ₇ -H), 4.0-5.0 (5H; m; pyrroline ring C ₅ -H ₂ , pyrroline ring C ₆ -H, C ₆ -H and C ₈ -H), 4.80 (HOD), 5.80 (1H, d, J = 1 Hz, C ₅ -H)

HPLC (holding time): 23.2 and 28.8 minutes

Example 30

(5R, 6S, 8R) -2-[(2,3-dihydro-1H-pyrazolo [1,2-a] pyrazolium-2-yl) thio-6- (1-hydroxyethyl) -2 -Phenem-3-carboxylate Synthesis

The target compound was recovered according to the method similar to Example 19, except that 2,3-dihydro-2-mercapto-1H-pyrazolo [1,2-a] pyrazolium trifluoromethanesulfonate was used. Obtained as a yellow powder.

IR (KBr disc) cm ₁₈ ; 1775, 1995

UVλmax (H ₂ O): 250, 323

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz, CH ₂ ), 4.03 (1H; d; J = ₂ , 6 Hz, CH ₃ ), 4.15-4.43 (1H; m; C ₈ -H ), 5.80 (1H; d; J = 2 Hz, C ₅ -H), 6.97 (1H; t; J = 3 Hz, pyrazollium-H, 6 position), 8.28 (2H; d; J = 3 Hz, pyrazollium- H, 5-position and 7 positions)

HPLC (retention time): 13 minutes.

Condition: Carrier: Nucleosil 7C ₁₈ (10 × 300 mm)

Solvent: 3% Acetonitrile-water

Flow rate: 5ml / min

Example 31

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(4-methylpyrrolidinium-1-yl) thio] -2-phenem-3-carboxylate (isomer A and B) Synthesis

p-nitrobenzyl (5R, 6S, 8R) -2-ethylsulfinyl-6- (1-hydroxyethyl) -2-phenem-3-carboxylate (150 mg) and 1-mercapto-4-methylpyrroli Zidinium, trifluoromethane sulfonate (215 mg) was dissolved in 4 ml of DMF, cooled to -40 ° C and 130 mg of diisopropylethylamine was added with stirring, followed by stirring for 1 hour in an argon atmosphere. Ether is added to the reaction mixture and the ether phase is removed. The residue is dissolved in a mixture of THF (20 ml) and phosphate buffer (pH 7.0; 20 ml) and 200 mg of platinum oxide is added and reduced for 1 hour under 4 atmospheres of hydrogen. The concentrate obtained by concentrating the water phase under reduced pressure was subjected to column chromatography (20 × 300 mm) chromatography using Diion HP-20, and the fractions eluted with 5% THF-water were collected and concentrated under reduced pressure, followed by HPLC treatment [carrier: nucleo] Thread 7C ₁₈ (10 × 300mm); menstruum; 5% acetonitrile-water; Flow rate: 5 ml / min. Lyophilization treatment affords the desired compound as an off-yellow powder.

Isomer A: Yield 15 mg

IR (KBr disc) cm ⁻¹ ; 1765, 1590

UVλmax (H ₂ O): 258, 323

⁺NCH₃), 3.40-4.60(8H;m), 4.80(HOD), 3.80(1H;d;J=2Hz;C₅-H)NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz, CH ₃ ), 1.90-2.90 (6H; m), 3.29 (3H; s;

⁺ NCH ₃ ), 3.40-4.60 (8H; m), 4.80 (HOD), 3.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 12 minutes.

Isomer B: Yield 22 mg

IR (KBr disc) cm ⁻¹ ; 1760, 1590

UVλ max (H ₂ O) nm: 258, 324

⁺NCH₃), 3.50-4.50(8H;m), 4.80(HOD), 5.80(1H;d;J=2Hz;C₅-H)NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz, CH ₃ ), 1.90-3.10 (6H; m), 3.32 (3H; s;

⁺ NCH ₃ ), 3.50-4.50 (8H; m), 4.80 (HOD), 5.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 17.5 minutes

Example 32

(5R, 6S, 8R) -2-[(4- (2-fluoroethyl) -pyrrolidinium-1-yl) thio] -6- (1-hydroxyethyl) -2-phenem-3- Carboxylate (Isomers A and B) Synthesis

The procedure similar to Example 31 was repeated using 4- (2-fluoroethyl) -1-mercapto-pyrrolidinium trifluoromethane sulfonate to afford the desired compound as an off-white powder.

Isomer A:

IR (KBr disc) cm ⁻¹ ; 1770, 1595

UVλmax (H ₂ O): 255, 323

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz; CH ₃ ), 1.90-2.80 (6H; m), 3.20-4.50 (10H; m), 4.80 (HOD), 5.20-5.40 (1H m), 5.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 10.5 minutes

Condition: Carrier: Nucleosil 7C ₁₈ (10 × 300 mm)

Solvent: 7% Acetonitrile-water

Flow rate: 5ml / min

Isomer B:

IR (KBr disc) cm ⁻¹ ; 1770, 1590

UVλmax (H ₂ O): 255, 323

NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz; CH ₂ ), 1.80-3.00 (6H; m), 3.50-4.50 (10H; m), 4.80 (HOD), 5.20-5.40 (1H m), 5.80 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (holding time): 12.5 minutes

Example 33

(5R, 6S, 8R) -6- (1-hydroxyethyl) -2-[(4-methylpyrrolidinium-2-yl) thio] -2-phenem-3-carboxylate (isomers A and B )synthesis

Except for using 2-mercapto-4-methylpyrrolidinium, trifluoromethane sulfonate, a similar procedure as in Example 31 was obtained to obtain the target compound as an off-yellow powder.

Isomer A:

IR (KBr disc) cm ⁻¹ ; 1765, 1590

UV lambda max (H ₂ O): 250, 322

⁺NCH₃), 3.55-3.80(3H;m), 3.90-4.45(5H;m), 4.80(HOD), 5.78(1H;d;J=2Hz;C₅-H)NMRδ (D ₂ O) ppm: 1.37 (3H; d; J = 6 Hz; CH ₃ ), 1.80-2.70 (6H; m), 3.30 (3H; s;

⁺ NCH ₃ ), 3.55-3.80 (3H; m), 3.90-4.45 (5H; m), 4.80 (HOD), 5.78 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (retention time): 11.5 minutes

Condition: Carrier: Nucleosil 7C ₁₈ (10 × 300 mm)

Solvent: 5% Acetonitrile-water

Flow rate: 5ml / min

Isomer B:

UVλ max (H ₂ O) nm: 250, 323

⁺NCH₃), 3.40-4.40(7H;m), 4.80(HOD), 5.76(1H;d;J=2Hz;C₅-H)NMRδ (D ₂ O) ppm: 1.36 (3H; d; J = 6 Hz; CH ₂ ), 1.80-2.60 (6H; m), 2.70-3.10 (1H; m), 3.24 (3H; s;

⁺ NCH ₃ ), 3.40-4.40 (7H; m), 4.80 (HOD), 5.76 (1H; d; J = 2 Hz; C ₅ -H)

HPLC (holding time): 15 minutes

Reference Example 12

Synthesis of 2,3-dihydro-1-mercapto-1H-indoliginium trifluoromethane sulfonate

(1) ethyl picolinoacetate:

Sodium hydride was dissolved in 200 ml of DMF, and a mixed solution of ethyl acetate (20 g) of ethyl acetate (17.5 g) dissolved in 150 ml of DMF at a bath temperature of 50 ° C. was added dropwise to the solution for about 30 minutes. The reaction solution was stirred at 50 ° C. for 30 minutes, cooled under ice cooling, and acetic acid (13.0 g) was added dropwise. The reaction solution was diluted with ethyl acetate, washed with water, dried over MgSO ₄ , and the solvent was distilled off in vacuo to yield the target compound (yield: 21.2 g)).

NMRδ (CDCl ₃ ) ppm: 1.23 (3H; t; J = 7 Hz), 4.16 (2H; s), 4.16 (2H; q; J = 7 Hz), 7.40 (1H; ddd, J = 1, 7, 7 Hz) , 7.77 (1H; dd; J = 1, Hz), 7.95 (1H; ddd, J = 1, 7, 7,7 Hz), 8.58 (1H; dd; J = 1,7 Hz)

(2) ethyl 3-hydroxy-3- (2-pyridyl) propionate:

The compound (21.0 g) obtained in step (1) was dissolved in 300 ml of ethanol, and sodium borohydride (2.1 g) was added while stirring under ice cooling, followed by stirring for 30 minutes at the same temperature. The reaction solution is packed with 0.5N hydrochloric acid under ice-cooling, and the solvent is distilled under vacuum. The residue is partitioned between water and ethyl acetate and ethyl acetate is washed with water, dried over MgSO ₄ and the solvent is distilled in vacuo. The residue was subjected to column chromatography treatment using 150 g of silica gel [eluent: benzene / ethyl acetate (1: 1 V / V)] to give the target compound (yield: 14.6 g).

NMRδ (CDCl ₃ ) ppm: 1.14 (3H; t; J = 7 Hz), 2.61 (1H; dd; J = 8, 17 Hz), 2.80 (1H; dd, J = 5, 17 Hz), 4.04 (2H; q; 7 Hz), 4.30 (1 H; m), 5.05 (1 H; m), 7.04 (1 H; dt; J = 1, 7 Hz), 7.27 (1 H; dd, J = 1, 7 Hz), 7.55 (1 H; dt; J = 1,7 Hz), 8.36 (1H; dd; J = 1,7 Hz)

(3) ethyl 3-benzoylthio-3- (2-pyridyl) propionate:

Triphenylphosphine (37.6 g) is dissolved in 400 ml of THF, diethyl azodicarboxylate (25.0 g) is added under ice cooling and stirring is continued for 30 minutes at the same temperature. The mixture (14.0 g) obtained in step (2) was added to thiobenzoic acid (16.0 g) and 100 ml of THF. A mixed solution was added to the reaction solution and stirred for 2 hours at the same temperature. The residue obtained by vacuum evaporation of the reaction solution is partitioned between ethyl acetate and an aqueous solution of 5% sodium hydrogen carbonate. The ethyl acetate layer is washed with water, dried over MgSO ₄ and the solvent is removed by vacuum distillation. The residue obtained by washing the precipitated crystals with isopropyl ether was chromatographed [eluent: benzene / ethyl acetate (10: 1 V / V)] on a column filled with 500 g of silica gel to give the target compound (yield: 15.2g).

NMRδ (CDCl ₃ ) ppm: 1.17 (3H; t; J = 7 Hz), 3.12 (1H; dd, J = 7, 15 Hz), 3.44 (1H; dd; J = 8, 15 Hz), 4.08 (2H; q; J = 7 Hz), 5.38 (1H; dd; J = 8,15 Hz), 7.1-7.8 (6H; m), 7.8-8.0 (2H <m), 8.56 (1H; dd; J = 1,7 Hz)

(4) ethyl 3-p-methoxybenzylthio-3 (2-pyridyl) propionate:

The compound (5.55 g) obtained in step (3) is dissolved in 80 ml of methanol, sodium methoxylate (0.95 g) is added under ice cooling, and the mixture is stirred at the same temperature for 2.5 hours. Acetic acid (1.06 g) is added to the reaction solution and the mixture is evaporated in vacuo. The residue is partitioned between 5% aqueous sodium bicarbonate solution and ethyl acetate, the ethyl acetate is dried over MgSO ₄ and the solvent is removed by vacuum distillation. The oily substance (5.02 g) obtained here was dissolved in 150 ml of benzene, and p-methoxybenzylchloride (2.76 g) and DBU (2.68 g) were added thereto and stirred at room temperature for 1 hour. The reaction solution was washed with water, dried over MgSO _4, and the residue obtained by vacuum distillation of the solvent was subjected to column chromatography using a column filled with 150 g of silica gel [eluent: benzene / ethyl acetate (10: 1 V / V)]. To obtain the target compound (yield: 3.26 g).

NMRδ (CDCl ₃ ) ppm: 1.12 (3H; t; J = 7 Hz), 2.88 (1H; dd, J = 7, 16 Hz), 3.13 (1H; dd; J = 9, 16 Hz), 3.60 (2H; s) , 3.76 (3H; s), 4.03 (2H; q; J = 7 Hz), 4.28 (1H; dd; J = 7, 9 Hz), 6.76 (2H; d; J = 9 Hz), 7,13 (2H; d ; J = 9 Hz), 7.0-7.4 (2H; m), 7.57 (1H; dt; J = 2,8 Hz), 8.48 (1H; dd; J = 2,7 Hz)

(5) 3-p-methoxybenzylthio-3- (2-pyridyl) propanol:

LiA ₁ H ₄ (1.08 g) was dissolved in 150 ml of ether, and the solution obtained by adding the compound (9.34 g) obtained in the step (4) was added dropwise to 70 ml of ether under an argon atmosphere, and the mixture was heated to reflux for 1.5 hours. A 20% aqueous ammonium chloride solution (50 ml) is carefully added to the reaction solution under ice-cooling and filtered through celite. The ether phase is washed with water and the solvent dried over MgSO ₄ is distilled under vacuum. The residue was subjected to column chromatography (eluent: ethyl acetate) using 70 g of silica gel to give the target compound (yield: 7.36 g).

NMRδ (CDCl ₃ ) ppm: 2.0-2.3 (2H; m), 3.56 (2H; s), 3.5-3.8 (2H; m), 3.78 (3H; s), 4.08 (1H; t; J = 7 Hz), 6.80 (2H; d; J = 9 Hz), 7.15 (2H; d; J = 9 Hz), 7.1-7.5 (2H; m), 7.68 (1H; dt; J = 2, 8 Hz), 8.54 (1H; dd; J = 2,7 Hz)

(6) 2,3-dihydro-1- (p-methoxybenzylthio) -1H-indoliginium p-toluenesulfonate:

The compound (7.86 g) obtained in step (5) was dissolved in 500 ml of pyridine, and p-toluenesulfonyl chloride (7.10 g) was added thereto, stirred for 4.5 hours under ice cooling, and added to 100 ml of water, followed by 1N NaOH Adjust the pH to 9.2 with urea and continue with ether to remove pyridine. The water phase is washed with benzene and ethyl acetate and the pH is adjusted to 1.8 by addition of concentrated hydrochloric acid followed by vacuum evaporation. Ethanol is added to the residue and the insolubles are filtered off. The filtrate is decolorized with activated charcoal and the filtrate is concentrated and placed in ether. The supernatant is removed and the precipitate is dried to give the desired compound (yield: 9.96 g).

NMRδ (DMSO-d ₆ ) ppm: 2.26 (3H; s), 2.2-2.6 (1H; m), 2.6-3.1 (1H, m), 3.70 (2H; s), 3.90 (3H, s), 3.9 ( 1H; m), 4.83 (2H; m), 6.82 (2H; d; J = 9 Hz), 7.06 (2H; d; J = 9 Hz), 7.23 (2H; d; J = 9 Hz), 7,46 (2H) d; J = 9 Hz), 7.8-8.1 (2H; m), 8.41 (1H; dt; J = 1,8 Hz), 8.91 (1H; dd; J = 1,7 Hz)

(7) 2,3-dihydro-1-mercapto-1H-indoliginium trifluoromethane sulfonate:

The compound (8.29 g) and anisole (10.14 g) obtained in the step (16) were dissolved in 50 ml of trifluoro acetic acid, trifluoromethanesulfonic acid (4.55 g) was added under ice cooling, followed by stirring at the same temperature for 30 minutes. And stir at room temperature for 1.5 hours. The reaction solution is evaporated in vacuo and the residue obtained is washed with hexane, isopropyl ether and ether, respectively. The residue is dissolved in methanol, poured into ether with stirring and the supernatant is removed. Precipitation is dried to give the target compound (yield: 4.20 g).

NMSO (DMSO-d ₆ ) ppm: 2.1-2.5 (1H; m), 2.6-3.2 (1H; m), 4.5-5.1 (3H; m), 7.9-8.2 (2H; m), 8.40 (1H; dt ; J = 1,8 Hz), 8.91 (1H; dd; J = 1,7 Hz)

Reference Example 13

2,3-dihydro-2-mercapto-1H-indoliginium trifluoromethane sulfonate

(1) ethyl 2-hydroxy-3- (2-pyridyl) propionate:

The product obtained by reacting 4.31 g of ethyl 2-pyridylpyruvate (synthesized according to the method shown in Chem. Pharm. Bull. 1972, 20, 1628) was similar to the method described in step (2) of Reference Example 12. Post-treatment according to the method affords the target compound (yield: 3.84 g).

NMRδ (CDCl ₃ ) ppm: 1.22 (3H; t; J = 7 Hz), 3.18 (1H; dd; J = 7, 14 Hz), 3.25 (1H; dd; J = 4, 14 Hz), 4.17 (2H; d; J = 7Hz), 4.60 (1H; dd; J = 4,7Hz), 4.87 (1H; brs), 7.0-7.2 (2H; m), 7.56 (1H; dt; J = 2,7,7Hz), 8.41 (1H; dd; J = 1,6 Hz)

(2) ethyl 2-benzoylthio-3- (2-pyridyl) propionate:

The target compound is obtained by reacting the compound (3.96 g) obtained in step (1) above and post-treating the product obtained according to a method similar to that in step (3) of Reference Example 12 (yield: 5.92 g).

NMRδ (CDCl ₃ ) ppm: 1.17 (3H; t; J = 7 Hz), 3.39 (1H; dd; J = 7, 15 Hz), 3.50 (1H; dd; J = 7, 15 Hz), 4.15 (2H; q; J = 7Hz), 4.88 (1H, t, J = 7Hz), 7.0-7.6 (6H; m), 7.86 (2H, m), 8.46 (1H, dd, J = 1,6Hz)

(3) ethyl 2-p-methoxybenzylthio-3- (2-pyridyl) propionate:

The product obtained by reacting the compound (5.90 g) obtained in the step (2) was worked up according to a method similar to that in step (4) of Reference Example 12 to obtain the target compound (yield: 4.76 g). ).

NMRδ (CDCl ₃ ) ppm: 1.21 (3H; t; J = 7 Hz), 3.10 (1H; dd; J = 7 Hz, 15 Hz), 3.27 (1H, d, J = 8, 15 Hz), 3.75 (3H, s) , 3.77 (2H, s), 3.7-3.9 (1H; m), 4.12 (2H, q, J = 7 Hz), 6.76 (2H, d, J = 9 Hz), 6.9-7.3 (4H, m), 7.50 ( 1H, dt, J = 2,8 Hz), 8.40 (1H, dd, J = 1,6 Hz)

(4) 2-p-methoxybenzylthio-3- (2-pyridyl) propanol:

The product obtained by reacting the compound obtained in step (3) was worked up according to a method similar to that in step (5) of Reference Example 12 to obtain the target compound (yield: 694 mg).

NMRδ (CDCl ₃ ) ppm: 3.0-3.5 (2H, m), 3.5-4.3 (3H, m), 3.64 (2H, s), 3.76 (3H, s), 6.76 (2H, d, J = 9Hz), 7.0-7.3 (2H, m), 7.12 (2H, d, J = 9Hz), 7.54 (1H, dt, 7Hz), 8.40 (1H, dd, J = 2,7Hz)

(5) 2,3-dihydro-2- (p-methoxybenzylthio) -1H-indoliginium chloride:

The compound (0.8 g) obtained in step (4) was dissolved in carbon tetrachloride (40 ml), and triphenylphosphine (1.45 g) was added thereto and heated to reflux for 20 hours. After cooling, the supernatant is removed by decantation and the separated blackish blue oil is partitioned between water and ethyl acetate. The water phase is washed five times with chloroform, decolorized with activated charcoal and then evaporated in vacuo to give the desired compound (yield: 0.53 g).

NMRδ (DMSO-d ₆ ) ppm: 3.2-3.7 (2H, m), 3.77 (3H, s), 3.97 (2H, s), 3.7-4.0 (1H, m), 4.83 (1H, dd, J = 5 , 13 Hz), 5.10 (1H, dd, J = 7,13 Hz), 6.92 (2H, d, J = 9 Hz), 7.32 (2H, d, J = 9 Hz), 7.8-8.2 (2H, m), 8.54 ( 1H, dt, J-1,8Hz), 9.00 (1H, dd, J = 1,7Hz)

(6) 2,3-dihydro-2-mercapto-1H-indoliginium trifluoromethanesulfonate

The desired compound was obtained by subjecting the product obtained by reacting the compound (535 mg) obtained in the step (5) to a method similar to that obtained in the step (7) of Reference Example 12 (yield: 520 mg). .

NMR δ (D ₂ O) ppm: 3.2-4.4 (3H, m), 4.7-5.0 (1H, m), 5.1-5.4 (1H, m), 7.7-8.1 (2H, m), 8.52 (1H, dt , J = 2,8 Hz), 8.84 (1H, dd, J = 2,8 Hz)

Reference Example 14

6,7-dihydro-7-mercapto-5H-pyrrolo [1,2-a] imidazole trifluoromethanesulfonate

(1) 6,7-dihydro-7-P-methoxybenzylthio-5H-pyrrolo [1,2-a] imidazole: 2-ethoxy-3-p-ethoxybenzylthio-1-py Raline (531 mg) (synthesized according to the method disclosed in Japanese Patent Laid-Open No. 56987/1985), iminoacetaldehydedimethylacetyl (315 mg) and acetic acid (180 mg) are dissolved in 30 ml of ethanol and heated to reflux for 1 hour. The residue obtained by vacuum evaporation of the reaction solution was partitioned between 40 ml of water containing 4 ml of 1N NaOH aqueous solution and benzene, the benzene phase was washed with water, dried over MgSO ₄ and the solvent was distilled under vacuum to give 700 mg of a colorless transparent oily substance. To obtain. This material is dissolved in 30 ml of benzene and heated to reflux for 1 hour with the addition of 1.14 g of p-toluenesulfonic acid (hydrate). The reaction solution is cooled, washed with 5% aqueous sodium bicarbonate solution and water, dried, and the solvent is evaporated. The residue was subjected to column chromatography (eluent: chloroform / methanol (19: 1 v / v)) using a column packed with 20 g of silica gel to give the title compound as a colorless transparent oil. The compound is crystallized (melting point: 65 to 75 ° C .; yield: 0.53 g) while standing in a freezer.

IR (KBr disc) cm ^-1 : 1600, 1500, 1240

NMR δ (CDCl ₃ ) ppm: 2.2-2.5 (1H, m), 2.6-3.0 (1H, m), 3.76 (3H, s), 3.7-4.2 (5H, m), 5.79 (2H, d, J = 9 Hz), 6.83 (1H, d, J = 2 Hz), 7.05 (1H, d, J = 2 Hz), 7.29 (2H, d, J = 9 Hz)

(2) 6,7-dihydro-7-mercapto-5H-pyrrolo [1,2-a] imidazoletrifluoromethanesulfonate: See compound (0.50 g) obtained in step (1) above. Post-treatment in a manner similar to that of step (7) of Example 12 gave the target compound (yield: 0.56 g).

NMR δ (DMSO + D ₂ O) ppm: 2.0-2.7 (1H, m), 3.0-3.6 (1H, m), 4.24 (2H, m), 4.70 (1H, dd, J = 4,7Hz), 7.65 (2H, brs)

Reference Example 15

6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-a] imidazole trifluoromethanesulfonate

(1) 4-benzoylthio-2-pyrrolidone: Triphenylphosphine (10.5 g) is dissolved in 50 ml of THF, and a THF solution (3 ml) of diethylazodiethyl carboxylate (6.97 g) is added under ice-cooling. . After 30 minutes, 30 ml of THF was added and 4-hydroxy-2-pyrrolidone (2.02 g) [synthesized according to the method better described by Pellegata et al., Synthesis, 615 (1978)] and thiobenzoic acid (4.71 ml) were THF. Suspended in 80ml dropwise for 20 minutes. Again, the solution was stirred for 1.5 hours at room temperature and then concentrated in vacuo. The concentrate is dissolved in ethyl acetate and then washed with 5% aqueous sodium hydrogen carbonate solution, then with saturated aqueous NaCl solution, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was chromatographed using 120 g of silica gel [eluted with a mixture of benzene and ethyl acetate. The mixing ratio was gradually changed from 1: 1 to 1: 4 and 1: 9] to obtain the target compound as a yellow powder (melting point: 110 to 112 ° C., yield: 3.47 g).

IR (KBr disc) cm ^-1 : 1690, 1650

NMR δ (CDCl ₃ ) ppm: 2.41 (1H, dd, J = 6,18 Hz), 2.93 (1H, dd, J = 9,18 Hz), 3.43 (1H, dd, J = 5,10 Hz), 3.99 (1H , dd, J = 8,10Hz), 4.4 (1H, m), 6.8 (1H, br), 7.4-8.0 (5H, m)

(2) 4- (p-methoxybenzylthio) -2-pyrrolidone: Compound sodium (0.37 g) was dissolved in 30 ml of methanol with ice cooling, cooled to -10 ° C, and then the compound obtained in step (1). (3.43 g) is added to methanol (30 ml) and the resulting solution is added and stirred at the same temperature for 1 hour. To this reaction solution p-methoxybenzyl chloride (2.51 g) is added at the same temperature and stirred at room temperature for 1.5 hours. The solvent is distilled under vacuum, washed with water by addition of ethyl acetate and then again with 5% aqueous solution of sodium bicarbonate, 10% aqueous solution of citric acid and saturated aqueous solution of NaCl. This solution is dried over Na ₂ SO ₄ and concentrated in vacuo. The evaporation residue is purified by passing through a column packed with 160 g of silica gel. First elute with benzene-ethyl acetate (1: 1 v / v) mixed solvent (the amount of ethyl acetate gradually increased) and then with pure ethyl acetate. Fractions containing the target compound are collected and concentrated to give the target compound as colorless crystals (melting point: 95 to 96 ° C; yield: 2.69 g).

IR (KBr disc) cm ^-1 : 3240, 1670, 1610, 1510

NMR δ (CDCl ₃ ) ppm: 2.16 (1H, dd, J = 7, 18 Hz), 2.64 (1H, dd, J = 8, 18 Hz), 3.2-3.6 (3H, m), 3.73 (2H, s), 3.80 (3H, s), 6.1 (1H, br), 6.87 and 7.27 (2H each, d, J = 9 Hz)

(3) 2-ethoxy-4- (p-methoxybenzylthio) -1-pyrroline: a meervine reagent prepared from epichlorohydrin (2.75 ml) and BF ₃ (C ₂ H ₅ ) ₂ O (Meerwein reagent: H. Meerwein et al., "Organic Synthesis", Coll, Vol, V, p.1080) was dissolved in 30 ml of anhydrous dichloromethane, and the compound (2.37 g) obtained in the above step (2) was added with ice cooling. And stir at room temperature for 1 hour. The solution is thickened by addition of a 5% aqueous solution of sodium carbonate and extracted with dichloromethane. The organic phase is washed with water, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue is purified by column chromatography treatment (eluent: benzene / ethyl acetate (1: 1 v / v)) using a column packed with 50 g of silica gel. The fractions containing the target compound were combined and concentrated in vacuo to give the target compound in the form of greyish brown oil (yield: 197 g).

NMR δ (CDCl ₃ ) ppm: 1.28 (3H, t, J = 7 Hz), 2.6 (2H, m), 3.4-3.9 (5H, m), 3.77 (3H, s), 4.17 (2H, q, J = 7 Hz), 6.82 and 7.21 (2H each, d, J = 9 Hz)

(4) 6,7-dihydro-6-p-methoxybenzylthio-5H-pyrrolo [1,2-a] imidazole: 2-ethoxy-4-p-methoxybenzylthio-1-py Raline (930 mg) and amino acetaldehyde dimethyl acetal (552 mg) are dissolved in 50 ml of methanol, acetic acid (315 mg) is added and heated to reflux for 1 hour. After cooling, ethyl acetate and water are added thereto and the solution is basified with 1N NaOH and partitioned between ethyl acetate and water. The ethyl acetate phase is washed with water and then with saturated aqueous NaCl solution, then dried over NaSO ₄ and concentrated in vacuo. The residue is dissolved in 70 ml of benzene and heated to reflux for 3 hours by addition of p-toluenesulfonic acid monohydrate (1 g). The solvent is evaporated in vacuo, ethyl acetate and water are added to the residue, the solution is basified with a 5% aqueous solution of sodium bicarbonate and partitioned. Ethyl acetate is washed with water, dried over NaSO ₄ and concentrated in vacuo. Purification by column chromatography using a column packed with 25 g of the residue silica gel [eluent: chloroform / methanol (19: 1 V / V /)], and the fractions containing the target compound were combined and concentrated in vacuo to give a brown oil. This target compound is obtained (yield: 731 mg).

NMR δ (CDCl ₃ ) ppm: 2.6-3.4 (2H, m), 3.5-4.2 (m), 3.77 (2H, s), 3.80 (3H, s), 6.79 and 7.02 (1H each, J = 1 Hz) , 6.85 and 7.24 (2H each, J = 9 Hz)

(5) 6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-a] imidazole trifluoromethanesulfonate:

The compound (354 mg) obtained in the step (4) was dissolved in a mixture of anisole (0.5 ml) and trifluoroacetic acid (4 ml), 0.12 ml of trifluoromethanesulfonic acid was added under ice cooling, and the temperature was raised to room temperature. And stir at this temperature for 30 minutes. The reaction solution was worked up according to a method similar to that of Reference Example 12- (7) to give the present desired compound in brown oil (yield: 390 mg).

NMR δ (D ₂ O) ppm: 3.1-3.9 (3H, m), 4.1-4.6 (2H, m), 4.80 (HOD), 7.43 (2H, s)

Reference Example 16

6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-c] imidazole trifluoromethanesulfonate synthesis:

(1) 3- (imidazol-4-yl) -2-p-methoxybenzylthio-propionic acid: 2-chloro-3- (imidazol-4-yl) propionic acid (6.01 g) prepared from L-histidine (see physiol. chem. 1942, 276, 126) is dissolved in 50 ml of water containing 3.0 g of NaOH, 5.3 g of p-methoxy benzylmercaptan is added and stirred at room temperature for 3 days. The reaction solution is washed with benzene, acetic acid is added to adjust the pH of the water phase to 3-4, and the water phase is concentrated under reduced pressure. The precipitated crystals are filtered and washed with ethanol and ether to give the target compound of colorless acicular crystals (melting point: 85-88 ° C .; yield 5.8 g).

IR (KBr disc) cm ^-1 : 3500-2500, 1580, 1510

N.M.R. δ (NaOD) ppm: 2.92 (2H, m), 3.43 (1H, dd, J = 7,8Hz), 3.70 (2H, s), 3.81 (3H, s), 6.82 (1H, d, J = 1Hz) , 6.93 (2H, d, J = 9 Hz), 7.27 (12H, d, J = 9 Hz), 7.58 (1H, d, J = 1 Hz)

(2) Cool 30 ml of methyl 3- (imidazol-4-yl) -2-p-methoxybenzylthiopropionate methanol to -10 ° C, add 0.47 ml of thiochloride dropwise with stirring, and then stir again for 10 minutes. . To this solution is added 1.46 g of the compound obtained in step (1) and stirred at room temperature for 1 hour. The solution is concentrated under reduced pressure, benzene is added to the residue and the solvent is distilled off under reduced pressure. A 5% aqueous solution of sodium bicarbonate is then added to the residue and extracted with chloroform. The extract is dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure to give the target compound (yield: 098 g).

NMR δ (CDCl ₃ ) ppm: 3.03 (2H, m), 3.54 (1H, dd, J = 6,8 Hz), 3.70 (2H, s), 3.79 (3H, s), 6.82 (3H, m), 7.22 (2H, d, J = 9Hz), 7.51 (1H, d, J = 1Hz)

(3) 3- (imidazol-4-yl) -2- (p-methoxybenzylthio) propanol: The compound (720 g) obtained in step (2) was dissolved in 15 ml of isopropanol and 780 mg of calcium chloride and 180 mg of NaBH ₄ were added. After addition, it is stirred for 5 hours at room temperature. To the reaction solution was added ethyl acetate, washed with water and saturated aqueous NaCl solution, dried over Na ₂ SO ₄ , column chromatography using a small amount of silica gel and distillation of the solvent under reduced pressure to obtain 566 mg of the target compound. .

NMR δ (CDCl ₃ ) ppm: 2.93 (3H, m), 3.64 (2H, m), 3.68 (2H, s), 3.77 (3H, s), 6.76 (1H, d, J = 1 Hz), 6.81 (2H , d, J = 9 Hz, 7.20 (2H, d, J = 9 Hz), 7.50 (1 H, d, J = 1 Hz)

Mass spectrometry (m / e): 279 (M ⁺ +1)

(4) 4- (2-chloro-3-p-methoxybenzylthiopropyl) imidazole: The compound (560 mg) obtained in the step (3) was dissolved in a mixed solvent of 20 ml of THF and 10 ml of carbon tetrachloride and triphenyl After adding 630 mg of phosphine, the mixture was heated to reflux for 2.5 hours. After cooling the solution, triethylamine was added and the solvent was distilled off under reduced pressure, and the residue obtained was purified by column chromatography using 15 g of silica gel and chloroform-methanol (9: 1 V / V) as eluent, followed by purification. 234 mg of the desired compound are obtained.

NMR δ (CDCl ₃ ) ppm: 2.79 (2H, d, J = 6 Hz), 3.14 (2H, m), 3.72 (2H, s), 3.78 (3H, s), 4.22 (1H, m), 6.82 (2H , d, J = 9 Hz, 6.87 (1 H, d, J = 1 Hz), 7.21 (2 H, d, J = 9 Hz), 7.56 (1 H, d, J = 1 Hz), 7.80 (1 H, brs)

(5) 6,7-dihydro-6-p-methoxybenzylthio-5H-pyrrolo [1,2-c] imidazole: The compound (2.21 g) obtained in the step (4) was added to 40 ml of acetone. Dissolve and add 11.1 g of sodium iodide and heat to reflux for 24 hours. The solvent is evaporated under reduced pressure, chloroform is added to the residue and washed with dilute aqueous solution of NaOH and saturated aqueous NaCl solution. The solution was dried over Na ₂ SO ₄ , the solvent was distilled off under reduced pressure, and the residue was purified by eluting with chloroform-methanol (9: 1 V / V) through a column filled with 60 g of silica gel to obtain 0.89 g of the target compound. To obtain.

NMR δ (CDCl ₃ ) ppm: 2.6-3.4 (2H, m), 3.77 (2H, s), 3.8 (3H, s), 3.5-4.25 (3H, m), 6.70 (1H, s), 6.86 (2H , d, J = 9Hz), 7.25 (2H, d, J = 9Hz), 7.42 (1H, s)

Mass spectrometry (m / e): 260 (M ⁺ )

(6) 6,7-dihydro-6-mercapto-5H-pyrrolo [1,2-c] imidazoletrifluoromethanesulfonate: Compound (323 mg) obtained in step (5) above anisole (0.4 ml) and trifluoroacetic acid (4 ml) are dissolved, 0.12 ml of trifluoromethanesulfonic acid is added under ice-cooling and stirred at room temperature for 30 minutes. After evaporation of the solvent under reduced pressure, the residue is added to xylene and the solvent is distilled off under reduced pressure and this operation is repeated several times. The residue obtained here is washed with ether to give 360 mg of the target compound.

TCL: Rf = 0.05 Silica gel plate (made by Merck). Thickness 0.25mm; Developing solvent: chloroform-methanol (1: 1 V / V mixture); Coloring purple with sodium nitroprusside.

NMR δ (D ₂ O) ppm: 2.9-3.9 (3H, m), 4.2-4.6 (2H, m), 4.80 (HOD), 7.24 (1H, s), 8.65 (1H, s)

Reference Example 17

6,7-dihydro-1-hydroxy-6-mercapto-5H-pyrrolo [1,2-c] imidazole:

(1) 1-t-butoxycarbonyl-2-ethoxycarbonyl-4-oxopyrrolidine: Ethyl ester of 1-t-butoxycarbonyl-L-hydroxy-proline (7.77 g) in DMSO (30 ml), dicyclohexylcarbodiimide (18.4 g) and 2.4 ml of pyridine are added followed by 1.2 ml of trifluoro acetic acid at room temperature with stirring. The mixture is stirred at the same temperature for 2 hours, then ethyl acetate is added and the insolubles are filtered off. The filtrate is washed with water, dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The residue was passed through a column packed with 120 g of silica gel to elute with benzene-ethyl acetate (98: 2 V / V) to afford the target compound (yield: 7.1 g).

NMR δ (CDCl ₃ ) ppm: 1.28 (3H, t, J = 6 Hz), 1.48 (9H, s), 2.40-3.13 (2H, m), 3.86 (2H, s), 4.18 (2H, q, J = 6 Hz), 4.65-4.86 (1 H, m)

(2) 1-t-butoxycarbonyl-2-ethoxycarbonyl-4-hydroxypyrrolidine: The compound (9.4 g) obtained in step (1) was dissolved in methanol (100 ml) and Na ₂ 0.7 g of BH ₄ are added at a temperature of 0-5 ° C. and stirred at the same temperature for 30 minutes. The solvent is then evaporated under reduced pressure and ethyl acetate, water and ammonium chloride are added to the evaporation residue to separate the organic phase. The organic phase was washed with water and saturated aqueous NaCl solution, dried over Na ₂ SO ₄ and distilled off the solvent under reduced pressure to give the target compound (yield: 9.4 g).

(3) 1-t-butoxycarbonyl-2-ethoxycarbonylpyrrolidin-4-yl, tosylate:

The compound (9.5 g) obtained in the previous step (2) was dissolved in pyridine (100 ml), cooled to 0 to 5 ° C., and then stirred with p-toluene sulfonyl chloride (10.5 g), and stirred at room temperature for 17 hours. Continue stirring. The solvent was distilled off under reduced pressure, ethyl acetate was added, washed with water, dried over Na ₂ SO ₄ and the solvent was distilled off under reduced pressure. The residue obtained here was passed through a column packed with 150 g of silica gel and eluted with a 95: 5 mixture of benzene and ethyl acetate to give the target compound (yield: 13.4 g).

NMRδ (CDCl ₃ ) ppm: 1.23 (3H; t; J = 6 Hz), 1.43 (9H; s), 2.20-2.53 (5H; m), 3.50-3.65 (2H; s), 3.95-4.45 (3H; m ), 4.86-5.10 (1H; m), 7.25 (2H; d; J = 9 Hz), 7.66 (2H; q; J = 9 Hz)

(4) 1-t-butoxycarbonyl-2-ethoxycarbonyl-4-p-methoxybenzylthiopyrrolidine:

13 g of the compound obtained in step (3) was dissolved in 300 ml of benzene, and a mixture of 9.7 g of p-methoxybenzylmercaptan and 9.6 g of DBU was heated and refluxed under an argon atmosphere for 7 hours. The solvent was evaporated under reduced pressure and the residue was purified by column chromatography using 300 g of silica gel and a mixture of 95: 5 of benzene and ethyl acetate as eluent to obtain the target compound (yield: 10.3 g).

NMRδ (CDCl ₃ ) ppm: 1.25 (3H; t; J = 6 Hz), 1.42 (9H; s), 1.70-2.60 (2H; m), 3.00-3.30 (2H; s), 3.67 (2H; s), 3.77 (3H, s), 4.00-4.30 (3H; m), 6.78 (2H; d; J = 9 Hz), 7.16 (2H; q; J = 9 Hz)

(5) 1-t-butoxycarbonyl-2-carbamoyl-4-p-methoxybenzylthio-pyrrolidine:

1.76 g of the compound obtained in the step (4) is dissolved in 30 ml of methanol, and 30 ml of a 30% aqueous ammonia solution is added, and stirred for 40 hours in a sealed tube at 40 to 45 ° C. The reaction mixture is extracted with ethyl acetate, washed with 10% aqueous citric acid solution and saturated aqueous NaCl solution, dried over Na ₂ SO _{4 and} the solvent is evaporated under reduced pressure. The residue was passed through a column packed with 30 g of silica gel, eluted with a 9: 1 mixture of chloroform and methanol to give 1.00 g of the target compound.

NMRδ (CDCl ₃ ) ppm: 1.45 (9H; s), 1.80-2.80 (2H; m), 3.20-3.70 (3H; m), 3.73 (2H; s), 3.79 (3H; m), 4.30 (1H; m), 6.84 (2H; d; J = 9 Hz), 7.23 (2H; d; J = 9 Hz)

(6) 2-carbamoyl-4-p-methoxybenzylthiopyrrolidine:

To a mixture of compound (1.00 g) and anisole (1.0 ml) obtained in step (5) is added 7 ml of trifluoroacetic acid and stirred at room temperature for 40 minutes. The solvent is evaporated under reduced pressure, then ether is added to the residue and washed. The residue is dissolved in dilute hydrochloric acid solution and washed with ether. The aqueous phase is added to a dilute aqueous NaOH solution and basified and extracted with chloroform. 0.59 g of the target compound is obtained as a white solid by drying over Na ₂ SO ₄ and evaporating the solvent under reduced pressure.

IR (KBr disc) ^-1 : 3400, 2800, 1650, 1515

NMRδ (CDCl ₃ ) ppm: 2.20 (2H; m), 2.90 (2H; m), 3.20 (1H; m), 3.69 (2H; s), 3.80 (3H; s), 3.90 (1H; m), 6.84 (2H; d; J = 9 Hz), 7.22 (2H; d; J = 9 Hz)

(7) 6,7-dihydro-1-hydroxy-6-p-methoxybenzylthio-5H-pyrrolo [1,2-c] imidazole

0.59 g of the compound obtained in step (6) was dissolved in 30 ml of ethanol, and 1.96 g of ethyl ester of ortho formic acid and 8 mg of p-toluenesulfonic acid hydrate were added and refluxed for 3 hours. The solvent-free residue was passed through a column packed with 20 g of silica gel, eluted with a mixture of chloroform-methanol (49: 1 V / V) to afford the desired compound (yield: 278 mg).

NMRδ (CDCl ₃ ) ppm: 2.60-3.35 (2H; m), 3.72 (2H; s), 3.80 (3H; s), 6.85 (2H; d; J = 9 Hz), 7.23 (2H; d; J = 9 Hz ), 8.16 (1 H; s)

(8) 6,7-dihydro-1-hydroxy-6-mercapto-5H-pyrrolo- [1,2-c] imidazole

To 358 mg of the product obtained in step (7), 0.5 ml of anisole and 4 ml of trifluoroacetic acid are added, and 0.1 ml of trifluoromethanesulfonic acid is further added, followed by stirring at room temperature for 3 hours. The solvent is evaporated under reduced pressure, xylene is added and then distilled under reduced pressure again and the process is repeated several times. The product obtained here was washed with petroleum ether and dried under reduced pressure to give the desired compound (yield: 390 mg).

TCL: Rf = 0.2

[Silica gel plate (Merck), 0.25mm thickness:

Developing solvent: CHCl ₃ : CH ₃ OH (9: 1 V / V); Coloring purple with sodium nitroprusside.

NMRδ (D ₂ O + DMSO d ₆ ) ppm: 2.9-3.35 (2H; m), 4.2-4.5 (2H; m), 7.25 (1H; s), 8.36 (1H; s)

Reference Example 18

6,7-dihydro-6-mercapto-5H-pyrrolo- [2,1-c] 1,2,4-triazole trifluoromethane sulfonate:

(1) 6,7-dihydro-6-p-methoxybenzylthio-5H-pyrrolo [1,2-c] 1,2,4-triazole:

2-methoxy-4-p-methoxybenzylthio-1-pyrroline (1.06 g) is added to ethanol (50 ml) and 0.36 g of formylhydrazine is added to the solution and heated to 80 ° C. under argon atmosphere with stirring. . After 1.5 hours, 0.24 g of formylhydrazine is added again and stirred while heating at the same temperature for 30 minutes. Then 0.6 g of acetic acid is added and stirred at the same temperature for 30 minutes and then cooled to room temperature. To the cooled product is added 5% aqueous solution of ethyl acetate and sodium carbonate to partition the product between these two liquids. The organic phase obtained here is washed with saturated aqueous NaCl solution, dried over Na ₂ SO ₄ and distilled off the solvent under reduced pressure to obtain the target compound.

Mass spectrometry (m / e): 279 (M ⁺ )

NMRδ (CDCl ₃ ) ppm: 2.3-3.0 (2H; m), 3.2 (2H; m), 3.7 (1H; m), 3.79 (3H; s), 6.83 (2H; d; J = 9 Hz), 7.21 ( 2H; d; J = 9Hz), 7.85 and 8.34 (0.5H each; S)

20 ml of acetic acid was added to the product obtained, heated at 100 ° C. for 4 hours, cooled, poured into ice water, neutralized with sodium carbonate, and extracted with ethyl acetate. The organic phase is washed, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue obtained here is then subjected to column chromatography using 20 g of silica gel and a mixture of CHCl ₃ and CH ₃ OH (9: 1 V / V) as eluent to give the title compound as a brown solid.

IR (KBr disc) cm ^-1 : 1600, 1520, 1500

NMRδ (CDCl ₃ ) ppm: 2.7-3.4 (2H; ddd), 3.80 (2H; s), 3.81 (3H; s), 3.8-4.2 (3H; m), 6.86 (2H; d; J = 9Hz), 7.24 (2H; d; J = 9 Hz), 8.02 (1H; s)

Mass spectrometry (m / e): 261 (M ⁺ )

(2) 6,7-dihydro-6-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole:

210 mg of the compound obtained in the step (1) was dissolved in a mixture of anisole (0.5 ml) and trifluoroacetic acid (4 ml), and 0.09 ml of trifluoromethanesulfonic acid was added under ice cooling and stirred at room temperature for 20 minutes. do. The reaction solution is concentrated under reduced pressure, 20 ml of xylene is added and concentrated again. Similarly, xylene is added and concentrated. Repeat this process twice. The residue is washed with ether with stirring and then ether is removed by decantation. The washing with ether was repeated twice and concentrated under reduced pressure to give the desired compound as a brown oil (yield: 250 mg).

NMRδ (D ₂ O) ppm: 3.1-4.0 (3H; m), 4.2-4.6 (2H; m), 9.23 (1H; s)

Reference Example 19

3-amino-6,7-dihydro-6-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethane sulfonate

(1) 4-p-methoxybenzylthio-2-thiosemicabozono pyrrolidine:

2-Ethoxy-4-p-methoxybenzylthio-1-pyrroline (900 mg) and thiosemicarbazide hydrochloride (433 mg) were added to ethanol (30 ml) and the solution was heated to reflux for 1.5 hours. The reaction solution is evaporated under reduced pressure and then the residue is partitioned between 5% aqueous solution of sodium hydroxide and ethyl acetate, the organic phase is dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The residue obtained here was passed through a column packed with 25 g of silica gel and eluted with ethyl acetate to give 230 mg of the target compound of colorless foam.

NMRδ (CDCl ₃ ) ppm: 2.3-3.0 (2H; m), 3.2-3.5 (2H; m), 3.72 (2H; s), 3.80 (3H; s), 3.70-4.0 (1H; m), 6.85 ( 2H; d; J = 9 Hz), 7.23 (2H; d; J = 9 Hz).

Mass spectrum analysis (m / e): 3.11 (M ⁺ +1)

(2) 4-p-methoxybenzylthio-2- (3-methylisothiosemicarbado) -1-pyrroline:

The compound (230 mg) prepared in the step (1) was dissolved in ethanol (20 ml), 312 mg of methyl iodide was added, followed by stirring at room temperature for 5 hours. The residue obtained by evaporation of the reaction solution under reduced pressure is partitioned between 30 ml of water containing 2 ml of 1N NaOH aqueous solution and ethyl acetate, the organic phase is washed, dried over Na ₂ SO ₄ and the solvent is distilled off. This gives 195 mg of the target compound of gray yellow oil.

NMRδ (CDCl ₃ ) ppm: 2.40 (3H; s), 2.48 (1H; dd; J = 8,16 Hz), 2.85 (1H; dd; J = 8,16 Hz), 3.2-3.7 (3H; m), 3.75 (2H; s), 3.80 (3H; s), 5.05 (2H; brs), 5.75 (1H; brs), 6.84 (2H; d; J = 9 Hz), 7.24 (2H; d; J = 9 Hz)

Mass spectrometry (m / e): 324 (M ⁺ )

(3) 3-amino-6,7-dihydro-6- (p-methoxybenzylthio) -5H-pyrrolo [2,1-c] -1,2,4-triazole:

190 mg of the compound obtained in step (2) was dissolved in a mixed solvent of ethanol (4 ml) and 1N NaOH aqueous solution, heated to reflux for 30 minutes, cooled, and the separated crystals were filtered and washed with water, ethanol and ether. 98 mg of the target compound are obtained.

Melting Point: 245-248 ° C. (Decomposition)

NMRδ (DMSO-d ₆ ) ppm: 2.7-4.1 (5H; m), 3.76 (3H; s), 3.81 (2H; s), 5.64 (1H; s), 6.88 (2H; d; J = 9Hz), 7.28 (2H; d; J = 9 Hz), 8.32 (1H; s)

Elemental Analysis (%): (C ₁₃ H ₁₆ N ₄ OS)

Theoretic: C; 56.50, H; 5.58, N; 20.27

Measured value: C; 56.31, H; 5.74, N; 20.00

(4) 3-amino-6,7-dihydro-6-mercapto-5H-pyrrolo [2,1-c] -1,2,4-triazole trifluoromethanesulfonate

According to a method similar to that described in Reference Example 12 [Step (7)], the compound (94 mg) obtained in Step (3) was treated and worked up to obtain 100 mg of the target compound.

NMRδ (D ₂ O) ppm: 2.8-3.1 (1H; m), 3.4-3.6 (1H; m), 3.8-4.0 (1H; m), 4.2-4.6 (2H; m), 4.80 (HOD)

Reference Example 20

2,3-dihydro-2-mercapto-1H-pyrazolo [1,2-c] pyrazollium trifluoromethane sulfonate

(1) 1-chloro-2-hydroxy-3-p-methoxybenzyl thiopropane:

2.78 ml of epichlorohydrin and 2.78 ml of p-methoxy benzyl mercaptan are heated at 120 to 130 ° C. for 3 hours with stirring. After cooling, the product was purified by column chromatography treatment with 100 g of silica gel and benzene as eluent to obtain 1.44 g of the target compound.

NMRδ (CDCl ₃ ) ppm: 2.40 to 2.80 (2H; m), 2.84 (1H; brs), 3.50 to 3.64 (2H; m), 3.70 (2H; s), 3.78 (3H; s), 6.85 (2H; d; J = 9 Hz), 7.25 (2H; d; J = 9 Hz)

(2) 1,2-ethoxy-3-p-methoxy benzylthiopropane:

To the compound (1.44 g) obtained in the step (1) was added a solution of water (1.8 ml) and NaOH (0.28 g) and stirred at room temperature for 17 hours. Dichloromethane is added to the reaction solution, washed, dried over NaSO _{4, and} the solvent is distilled off under reduced pressure. The residue was purified by passing through a column packed with 20 g of silica gel, eluting with chloroform to yield 0.54 g of the target compound.

NMRδ (CDCl ₃ ) ppm: 2.45 to 2.85 (4H; m), 2.96 to 3.20 (1H; m), 3.76 (2H, s), 3.79 (3H; s), 6.85 (2H; d; J = 9 Hz), 7.27 (2H; d; J = 9 Hz)

(3) 1- (2-hydroxy-3-p-methoxybenzylthiopropyl) pyrazole:

Compound (500 mg) and pyrazole (190 mg) obtained in step (2) were dissolved in DMF (2.5 ml), 655 mg of potassium carbonate was added and stirred at 90 ° C. for 7 hours. The solvent was distilled off under reduced pressure and the residue was purified by eluting with chloroform-methanol (99: 1 V / V) through a column filled with 20 g of silica gel to give 660 mg of the target compound.

NMRδ (CDCl ₃ ) ppm: 2.32 to 2.52 (2H; m), 3.70 (2H; s), 3.78 (3H; s), 3.80 to 4.28 (3H; m), 6.26 (1H; t; J = 3 Hz), 6.82 (2H; d; J = 9 Hz), 7.20 (2H; d; J = 9 Hz), 7.41, and 7.53 (each 1H; each d; J = 3 Hz)

(4) 1- (3-chloro-2-p-methoxybenzylthiopropyl) pyrazole:

It is dissolved in 6 ml of the compound 600 mg carbon tetrachloride prepared in the step (3), and 680 mg of triphenylphosphine is added, followed by stirring at 50 to 60 ° C. for 17 hours. After evaporation of the solvent under reduced pressure, the residue was purified by column chromatography using silica gel 15 g and chloroform as eluent to obtain 480 mg of the target compound.

NMRδ (CDCl ₃ ) ppm: 3.10 to 3.63 (5H; m), 3.79 (3H; s), 4.10 to 4.60 (2H; m), 6.26 (1H; t; J = 3 Hz), 6.82 (2H; d; J = 9 Hz), 7.20 (2H; d; J = 9 Hz), 7.45 and 7.56 (each 1H; each d; J = 3 Hz)

(5) 2,3-dihydro-2-p-methoxy benzylthio-1H-pyrazolo [1,2-a] pyrazolium iodide:

The compound (0.46 g) obtained in step (4) was dissolved in acetone (40 mg), and 1.3 g of potassium iodide was added, followed by heating to reflux for 30 hours. The solvent is distilled off under reduced pressure, ether is added to the residue obtained here, ether phase is removed, and chloroform is added to the resultant precipitate, followed by filtration to remove the insoluble matter. The filtrate is concentrated under reduced pressure to give the target compound (yield: 0.28 g).

NMRδ (CDCl ₃ ) ppm: 3.82 (H; s), 3.93 (2H; m), 4.35 to 4.70 (3H; m), 5.00 to 5.35 (2H; m), 6.70 to 7.00 (3H; m), 7.33 ( 2H; m), 8.37 (2H; d; J = 3 Hz)

(6) 2,3-dihydro-2-mercapto-1H-pyrazolo [1,2-a] pyrazolium trifluoro methanesulfonate:

The compound (0.28 g) obtained in step (5) was dissolved in a mixture of anisole (0.5 ml) and trifluoroacetic acid (4 ml), and 0.5 ml of trifluoromethane sulfonate was added, followed by stirring at room temperature for 1 hour. . After distilling off the solvent under reduced pressure, the process of adding xylene to the residue and concentrating the solution under reduced pressure was repeated several times. The residue is washed with petroleum ether to afford the desired compound (yield: 0.2 g).

NMRδ (D ₂ O) ppm: 4.40 to 4.70 (3H; m), 4.80 (HOD), 6.92 (1H; t; J = 3 Hz), 8.26 (2H; d; J = 3 Hz),

Reference Example 21

1-mercapto-4-methylpyrrolidinium trifluoromethane sulfonate synthesis:

(1) ethyl 3- (1-t-butoxycarbonyl pyrrolidin-2-yl) -3-inhibitor-propionate:

0.57 g of magnesium is added to 9 ml of ethanol, and 1.5 ml of carbon tetrachloride is added thereto, followed by stirring at room temperature for 2 hours. To the reaction mixture was added a solution of THF (30 ml) and monoethyl malonate (6.75 g), and the solvent was distilled off under reduced pressure when the reaction solution became clear. N-butoxycarbonylproline (6.45 g) was added to 5.35 g of carbonyldiimidazole while stirring and dissolving in THF (50 ml) at room temperature, followed by stirring for 30 minutes at the same temperature. To this reaction mixture is added a THF solution of the magnesium complex obtained above and stirred at room temperature for 2 hours. Ethyl acetate was added to the reaction solution, washed with 1N HCl, saturated aqueous sodium hydrogen carbonate solution and water, dried over NaSO ₄ , and the solvent was distilled off under reduced pressure. The residue obtained here was passed through a column packed with 100 g of silica gel and eluted with a mixture of benzene and ethyl acetate (95: 5 v / v) to give the purified desired compound (yield: 6 g).

NMRδ (CDCl ₃ ) ppm: 1.26 (3H; t; J = 6 Hz), 1.45 (9H; s), 1.70 to 2.30 (4H; m), 3.30 to 3.63 (4H; m), 4.16 (2H; q; J = 6 Hz), 4.10 to 4.45 (1 H; m)

(2) ethyl 3- (1-t-butoxycarbonylpyrrolidin-2-yl) -3-hydroxypropionate:

The compound (3 g) obtained in the previous step (1) was dissolved in methanol (30 ml), cooled to 0 to 5 ° C., then added with 200 mg of NaBH _{4 with} stirring and stirred at room temperature for 1 hour. To this reaction mixture, 2 g of ammonium chloride and water were added, extracted with chloroform, washed with saturated aqueous NaCl solution, dried over Na ₂ SO ₄ , and the solvent was distilled off under reduced pressure. The residue obtained here was passed through a column packed with 50 g of silica gel, eluted with chloroform-methanol (99: 1 v / v) to give the purified desired compound (yield: 3 g).

NMRδ (CDCl ₃ ) ppm: 1.26 (3H; t; J = 6 Hz), 1.47 (9H; s), 1.60 to 2.10 (4H; m), 2.33 to 2.53 (2H; m), 3.10 to 3.60 (2H, m ), 3.75 to 4.10 (1H; m), 4.13 (2H; q; J = 6 Hz)

(3) ethyl β- (1-t-butoxycarbonylpyrrolidin-2-yl) acrylate:

The compound (2.87 g) obtained in the above step (2) is dissolved in pyridine (20 ml) and 3.9 g of p-toluene sulfonyl chloride is added with stirring under ice cooling and stirred at room temperature for 17 hours. After stirring for another hour at 60 to 70 ° C., the solvent was distilled off, ethyl acetate was added to the residue, washed with water, dried over Na ₂ SO ₄ , and the solvent was distilled off under reduced pressure. The residue obtained here was purified by column chromatography using 50 g of silica gel and a mixed solvent of benzene and atel acetate (95: 5 V / V) to obtain a crude tosyl derivative of the target compound. This tosyl derivative is dissolved in 50 ml of chloroform and heated to reflux for 20 minutes after adding 1.5 g of DBU. The solvent is then distilled off under reduced pressure and ethyl acetate is added to the residue, followed by washing in the order of water, 0.5N HCl and saturated aqueous NaCl solution and dried over Na ₂ SO ₄ . The desired compound is obtained by distilling off the solvent under reduced pressure (yield: 2 g).

NMRδ (CDCl ₃ ) ppm: 1.26 (3H; t; J = 6 Hz), 1.43 (9H; s), 1.60 to 2.20 (4H; m), 3.30 to 3.55 (2H; m), 4.16 (2H; q; J = 6 Hz), 5.77 (1H; d; J = 15 Hz), 6.77 (1H; dd; J = 6.15 Hz).

(4) ethyl 3- (1-t-butoxycarbonyl pyrrolidin-2-yl) -3-p-methoxybenzylthio propionate:

Compound (2 g) obtained in the step (3) was dissolved in chloroform (30 ml), 1.6 g of p-methoxy benzyl mercaptan and 1.6 g of DBU were added, followed by stirring at room temperature for 17 hours. The residue obtained by distilling off the solvent under reduced pressure was passed through a column filled with 50 g of silica gel, and eluted with benzene-ethyl acetate (98: 2 V / V) to obtain the target compound (yield: 2.52 g).

NMRδ (CDCl ₃ ) ppm: 1.23 (3H; t; J = 6 Hz), 1.49 (9H; s), 1.65 to 2.10 (4H; m), 2.42 to 2.66 (2H; m), 3.10 to 3.70 (3H; m ), 3.74 (5H; s), 3.96 to 4.30 (3H, m), 6.75 (2H; d; J = 9 Hz), 7.10 to 7.30 (2H; m)

(5) ethyl 3- (1-t-butoxycarbonylpyrrolidin-2-yl) -3-p-methoxybenzylthiopropanol:

The compound (2.52 g) obtained in step (4) is dissolved in THF (40 ml), 0.2 g of NaBH ₄ is added under ice cooling, and the mixture is stirred at room temperature for 17 hours. To this reaction mixture ammonium chloride and water are added under ice cooling with stirring, followed by extraction with ethyl acetate. The organic phase is separated, washed with saturated NaCl solution, dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The residue was purified by column chromatography using 20 g of silica gel and chloroform as eluent to afford 2.14 g of the target compound.

NMRδ (CDCl ₃ ) ppm: 1.47 (9H; s), 1.50 to 2.20 (7H; m), 3.10 to 3.80 (6H; m), 3.74 (3H; s), 3.80 to 4.20 (1H, m), 6.76 ( 2H; d; J = 9 Hz), 7.10 to 7.30 (2H; m)

(6) 3- (1-t-butoxycarbonylpyrrolidin-2-yl) -3-p-methoxybenzylthiopropyl tosylate:

The compound (2.64 g) obtained in step (5) is dissolved in 30 ml of pyridine and 1.98 g of p-toluenesulfonyl chloride is added under ice-cooling and stirred at room temperature for 17 hours. The solvent is distilled off under reduced pressure, ethyl acetate is added to the residue, washed with water, dried over Na ₂ SO ₄ and the solvent is distilled off under reduced pressure. The residue obtained was passed through a column packed with 40 g of silica gel, eluted with a mixed solvent of benzine-ethyl acetate (98: 2 v / v) to give 2.68 g of the target compound.

NMRδ (CDCl ₃ ) ppm: 1.47 (9H; s), 1.50 to 2.20 (6H; m), 2.43 (3H; s), 3.10 to 3.73 (5H; m), 3.77 (3H; s), 3.80 to 4.30 ( 3H, m), 6.75 (2H; d; J = 9 Hz), 7.03-7.40 (4H; m), 7.72 (2H; d; J = 9 Hz)

(7) 3-p-methoxy benzylthio-3- (pyrrolidin-2-yl) -propyltosylate trifluoro acetate:

To 2.68 g of the compound prepared in step (6), 3 ml of anisole was added, and 10 ml of trifluoroacetic acid was added while stirring under ice cooling, followed by further stirring at the same temperature for 30 minutes. 2.7 g of the target compound are obtained by washing the residue obtained by distilling off the solvent under reduced pressure with petroleum ether and removing the solvent.

NMR δ (CDCl ₃ ) ppm: 1.45 to 2.30 (6H; m), 2.42 (3H; s), 2.70 to 3.60 (4H; m), 3.63 (2H; s), 3.77 (3H; s), 4.00 to 4.25 (2H; m), 6.75 (2H; d; J = 9 Hz), 7.02-7.40 (4H; m), 7.73 (2H; d; J = 9 Hz).

(8) 1-p-methoxy benzylthio pyrrolidinine:

The compound (2.7 g) obtained in step (7) is dissolved in DMF (20 ml), and 1.73 g of potassium carbonate is added and stirred at room temperature for 17 hours. To the residue obtained by stirring the solvent at 80 DEG C for 2 hours, distilled under reduced pressure, chloroform was added, washed with water, dried over Na ₂ SO ₄ , and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography using 15 g of silica gel and a mixed solvent of chloroform-methanol (95: 5 v / v) to give the target compound (yield: 0.9 g).

NMR δ (CDCl ₃ ) ppm: 1.50 to 2.30 (6H; m), 2.30 to 3.60 (6H; m), 3.64 (2H; s), 3.74 (3H; s), 6.75 (2H; d; J = 9 Hz) , 7.15 (2H; d; J = 9 Hz)

(9) 1-p-methoxy benzylthio-4-methylpyrrolidinium iodide:

The compound (260 mg) obtained in step (8) is dissolved in 5 ml of acetone and 710 mg of methyl iodide is added and left at room temperature for 48 hours. The reaction mixture is concentrated under reduced pressure and the residue obtained is washed with ether to give the title compound (yield: 400 mg).

(10) 1-mercapto-4-methylpyrrolidinium trifluoromethane sulfonate:

0.7 ml of anisole and 4 ml of trifluoro acetic acid were added to the compound (400 mg) obtained in the above step (9), 0.5 ml of trifluoro methane sulfonic acid was added dropwise with stirring at room temperature, followed by another 30 minutes at the same temperature. . The residue obtained by evaporating off the solvent is dispersed in xylene and distilled under reduced pressure. (Repeat this operation three times). The residue is washed with ether and dried to afford 300 mg of the desired compound.

NMR δ (D ₂ O) ppm: 1.90 to 2.80 (6H; m), 3.23 and 3.28 (3H; s), 3.30 to 4.15 (6H; m), 4.80 (HOD)

Reference Example 22

4-2- (fluoroethyl) -1-mercapto pyrrolidinium trifluoromethane sulfonate:

(1) 4- (2-fluoroethyl) -1-p-methoxy benzylthio pyrrolidinium bromide:

(1-p-methoxy benzylthio pyrrolidinine (263 mg), the product obtained in step (8) of Reference Example 21, was dissolved in acetone (5 ml) and 640 mg of 1-bromo-2-fluoroethane was added thereto. It is left for 4 days: The residue obtained by distilling off the solvent under reduced pressure is washed with ether and dried to give the target compound (yield: 390 mg).

NMR δ (CDCl ₃ ) ppm: 1.60 to 2.40 (6H; m), 2.80 to 4.20 (13H; m), 4.60 to 4.80 (1H; m), 5.15 to 5.35 (1H; m), 6.84 (2H; d; J = 9 Hz), 7.20 to 7.40 (2H; m).

(2) 4- (2-fluoroethyl) -1-mercaptopyrrolidinium trifluoromethane sulfonate:

0.7 ml of anisole and 4 ml of trifluoroacetic acid were added to the compound (390 mg) obtained in the previous step (1), 0.5 ml of trifluoromethane sulfonic acid was added dropwise while stirring at room temperature, and then again at the same temperature for 30 minutes. Stir. The solvent is distilled off under reduced pressure, xylene is added to the obtained residue, and the solvent is distilled under vacuum again. The residue is washed with ether and dried to give 330 mg of the desired compound.

NMR δ (D ₂ O) ppm: 1.80 to 2.80 (6H; m), 3.20 to 4.20 (8H; m), 4.80 (HOD), 5.10 to 5.30 (1H; m).

Reference Example 23

2-mercapto-4-methyl-pyrrolidinium trifluoromethane sulfonate:

(1) 1-t-butoxycarbonyl-4-p-methoxybenzylthio pyrrolidin-2-yl-methanol:

The compound (4.3 g) obtained in step (4) of Reference Example 17 was dissolved in 50 ml of THF, and 0.2 g of NaBH ₄ was added and stirred at room temperature for 17 hours. Water and ammonium chloride are added and extracted with ethyl acetate. After drying over Na ₂ SO ₄ , the solvent is distilled off under reduced pressure. The residue obtained here was passed through 60 g of silica gel and eluted with chloroform to give the purified desired compound (yield: 3.8 g).

NMR δ (CDCl ₃ ) ppm: 1.46 (9H; s), 1.80 to 2.10 (2H; m), 3.06 to 3.65 (5H; m), 3.68 (2H; s), 3.78 (3H; s), 3.90 to 4.30 (1H; m), 6.78 (2H; d; J = 9 Hz), 7.18 (2H; d; J = 9 Hz).

(2) 2-ethoxycarbonyl-3- (1-t-butoxycarbonyl-4-p-methoxybenzylthiopyrrolidin-2-yl) propionate:

Triphenyl phosphine (1.2 g) is dissolved in THF (9 ml) and 0.78 g of diethyl azodicarboxylate is added with stirring under ice cooling, followed by another 30 minutes at the same temperature. To this reaction solution was added a solution of compound (1.06 g) obtained in step (1), ethyl cyanoacetate (0.34 g), and a THF (6 ml) solution of 2,6-di-t-butyl phenol (0.63 g). Stir at room temperature for 2 hours. The residue obtained by distillation of the solvent under reduced pressure was purified by column chromatography using silica gel 40 g and benzene-ethyl acetate (95: 5 v / v) to obtain 0.65 g of the target compound.

NMR δ (CDCl ₃ ) ppm: 1.32 (3H; t; J = 6 Hz), 1.45 (9H; s), 1.80 to 2.23 (4H; m), 3.00 to 3.60 (4H; m), 3.72 (2H; s) , 3.80 (3H; s), 3.93 to 4.40 (3H; m), 6.84 (2H; d; J = 9 Hz), 7.23 (2H; d; J = 9 Hz).

(3) 3- (1-t-butoxycarbonyl-4-p-methoxybenzylthio pyrrolidin-2-yl) -propionitrile:

The compound (4.55 g) obtained in the step (2) is dissolved in 11 ml of ethanol, 15 ml of 1N NaOH is added at 0-5 ° C., and stirred at room temperature for 1 hour. To this solution is added water, washed with ether and the water phase is acidified with HCl and extracted with ethyl acetate. The extract was dried over Na ₂ SO ₄ and the residue obtained by distillation of the solvent under reduced pressure was dissolved in 50 ml of xylene and heated to reflux for 2 hours. The residue obtained by distillation of the solvent under reduced pressure was purified by passing through a column packed with 50 g of silica gel, eluting with benzene-ethyl acetate (95: 5 v / v) to obtain 3.4 g of the target compound.

NMR δ (CDCl ₃ ) ppm: 1.46 (9H; s), 1.50 to 2.50 (6H; m), 2.90 to 3.60 (3H; m), 3.68 (2H; s), 3.76 (3H; s), 3.80 to 4.30 (1H; m), 6.78 (2H; d; J = 9 Hz), 7.18 (2H; d; J = 9 Hz).

(4) 3- (1-t-butoxycarbonyl-4-methoxy benzylthio pyrrolidin-2-yl) propionic acid:

To the compound (3.4 g) obtained in step (3), a mixture solution of 50% aqueous ethanol solution (30 ml) and KOH (4.5 g) was added and heated to reflux for 6 hours. Water was added to the residue obtained by distillation of the solvent under reduced pressure, acidified with HCl, and extracted with ethyl acetate. The solvent of the extract is dried over Na ₂ SO ₄ and then distilled under reduced pressure. The residue obtained here was purified by column chromatography using silica gel 25 g and chloroform-methanol (99: 1 v / v) as eluent to obtain 2.9 g of the target compound.

NMR δ (CDCl ₃ ) ppm: 1.46 (9H; s), 1.50 to 2.15 (4H; m), 2.15 to 2.45 (2H; m), 3.00 to 3.60 (3H; m), 3.68 (2H; s), 3.78 (3H; s), 6.75 (2H; d; J = 9 Hz), 7.15 (12H; d; J = 9 Hz), 9.30 (1H; brs).

(5) 3- (1-t-butoxycarbonyl-4-p-methoxy benzylthiopyrroline-2-yl) propanol:

The compound (2.9 g) obtained in step (4) was dissolved in THF (30 ml), 1.13 ml of triethyl amine was added, cooled to -10 DEG C and 0.77 g of methyl chlorocarbonate was added dropwise while stirring. The mixture is stirred at 0-5 ° C. for 30 minutes and then again at 50 ° C. and the resulting precipitate is filtered off. 0.15 g of LiBH ₄ is added to the filtrate and stirred for 2 hours at room temperature. The reaction solution is concentrated under reduced pressure, ethyl acetate, water and ammonium chloride are added, the organic phase is separated, washed with saturated aqueous NaCl solution, dried over Na ₂ SO ₄ , and the solvent is evaporated to remove 2.6 g of the target compound. .

NMR δ (CDCl ₃ ) ppm: 1.46 (9H; s), 1.50 to 2.20 (6H; m), 3.00 to 3.70 (5H; m), 3.70 (2H; s), 3.78 (3H; s), 6.79 (2H d; J = 9 Hz), 7.16 (12H; d; J = 9 Hz).

(6) 3- (1-t-butoxycarbonyl-4-p-methoxy benzylthiopyrrolidin-2-yl) propyl tosylate:

The compound (2.6 g) obtained in step (5) is dissolved in 20 ml of pyridine, 2.8 g of p-toluene sulfonyl chloride is added, and then stirred at 0 to 5 ° C. for 17 hours. The solvent is distilled off under reduced pressure, and ethyl acetate is added to the residue and washed with water. The organic phase was dried over Na ₂ SO ₄ and the residue obtained by distillation under reduced pressure of the solvent in the organic phase was purified by column chromatography using 50 g of silica gel and benzene-ethyl acetate (98: 2 v / v) as eluent. 1.4 g of the desired compound is obtained.

NMR δ (CDCl ₃ ) ppm: 1.42 (9H; s), 1.40 to 2.00 (6H; m), 2.42 (3H; s), 2.80 to 3.60 (3H; m), 3.67 (2H; s), 3.76 (3H s), 3.85-4.23 (3H; m), 6.78 (2H; d; J = 9 Hz), 7.10-7.35 (4H; m), 7.72 (2H; d; J = 9 Hz).

(7) 2-p-methoxy benzylthio pyrrolidinine:

To the mixture of compound (1.4 g) and anisole (1.5 ml) obtained in step (6) was added 5 ml of trifluoro acetic acid with stirring under ice cooling, followed by stirring at the same temperature. The solvent was distilled off under reduced pressure, and the residue was washed with ether, dissolved in DMF (10 ml), and 0.95 g of potassium carbonate was added and stirred at room temperature for 17 hours. The solvent is distilled off under reduced pressure, washed with water by addition of chloroform, dried over Na ₂ SO ₄ , and the solvent is distilled off under reduced pressure. The residue was passed through a column packed with 15 g of silica gel to elute with chloroform-methanol (95: 5 v / v) to afford 0.47 g of the desired compound of interest.

NMR δ (CDCl ₃ ) ppm: 1.50 to 2.20 (6H; m), 2.26 to 2.60 (1H; m), 2.80 to 3.30 (4H; m), 3.69 (2H; s), 3.78 (3H; s), 6.78 (2H; d; J = 9 Hz), 7.18 (2H; d; J = 9 Hz).

(8) 2-p-methoxy benzylthio-4-methyl pyrrolidinium iodide:

400 mg of the compound obtained in the above step (7) is dissolved in 7 ml of acetone, 0.47 ml of methyl iodide is added, and left at room temperature for 2 days. The residue obtained by distillation of the solvent under reduced pressure was washed several times with ether and dried to obtain 600 mg of the target compound.

(9) 2-mercapto-4-methyl pyrrolidinium trifluoro methane sulfonate:

To a mixture of the compound (600 mg) anisole (1 ml) and trifluoro acetic acid (6 ml) obtained in step (8) was added 0.7 ml of trifluoro methane sulfonic acid with stirring at room temperature, followed by another 1 hour at the same temperature. While stirring. The residue obtained by distillation of the solvent under reduced pressure was washed repeatedly with petroleum ether to give the present desired compound as an oily substance (yield: 450 mg).

NMR δ (CDCl ₃ ) ppm: 1.70 to 2.60 (6H; m), 3.23 (3H; s), 3.25 to 4.40 (6H; m), 4.80 (HOD).

Claims (12)

Reacting a compound of formula (II) with a thiol compound of formula (III); Optionally removing the protecting group, alkylating the product or alkylating the reaction product, followed by removing the protecting group.

Wherein R ₁ is hydrogen, C ₁ to C ₆ alkyl or hydroxyl C ₁ to C ₆ alkyl group; COOR ₂ is a carboxyl group or carboxylate anion; R ₂ may be an ester moiety which may simultaneously act as a protecting group; R ₃ is a substituted or unsubstituted bicyclo heterocyclic group of formula T18;

Is the same ester residue as defined for R ₂ ; R ₅ is alkyl, aryl or aralkyl; Where m is 1,2 or 3; Q is T19; General formula

The partial structure of R ₃ represented by is a 5- or 6-membered nitrogen-containing heterocycle ring or a quaternary nitrogen-containing heterocycle ring, wherein the nitrogen-containing heterocycle ring is 1-4 selected from oxygen, sulfur and nitrogen. A saturated or unsaturated heterocycle ring having a hetero atom; R ₄ is halogen, amino, hydroxyl, cyano, optionally substituted C ₁ to C ₆ alkoxyl, optionally substituted carbamoyl, optionally substituted C ₁ to C ₆ alkyl, optionally substituted C ₂ to C ₆ alkenyl , Optionally substituted C ₂ to C ₆ alkynyl, optionally substituted C ₃ to C ₆ cycloalkyl, optionally substituted C ₃ to C ₆ cycloalkenyl, optionally substituted C ₃ to C ₆ cycloalkyl-C ₁ to C ₆ alkyl, optionally substituted C ₃ to C ₆ cycloalkenyl-C ₁ to C ₆ alkyl, optionally substituted C ₃ to C ₆ cycloalkenyl-C ₂ to C ₆ alkenyl, optionally substituted heterocyclyl, optionally Substituted heterocyclyl-C ₁ to C ₆ alkyl, optionally substituted heterocyclyl-C ₂ to C ₆ alkenyl, optionally substituted heterocyclyl-C ₂ to C ₆ alkynyl, optionally substituted C ₃ to C ₆ cycloalkylidene-C ₁ to C ₆ alkyl, optionally substituted C ₃ to C ₆ heterosa Icylidene-C ₁ to C ₆ alkyl, optionally substituted aryl, optionally substituted aryl-C ₁ to C ₆ alkyl, optionally substituted aryl-C ₂ to C ₆ alkenyl, optionally substituted aryl-C ₂ to C ₆ alkynyl, or a group of the general formula R _44- (CH ₂ ) vZ- (CH ₂ ) w-; Wherein Z is oxygen, sulfur SO, SO ₂ or NH; v is 0, 1 or 2; w is 0, 1, 2 or 3; R ₄₄ is hydrogen or C ₁ to C ₆ alkyl, C ₂ to C ₆ alkenyl, C ₃ to C ₆ alkynyl, C ₃ to C ₆ cycloalkyl, C ₃ to C ₆ cycloalkenyl, aryl, heterocycle A moiety selected from aryl, heterocyclyl-C ₁ to C ₆ alkyl, heterocyclyl-C ₂ to C ₆ alkenyl, and heterocyclyl-C ₂ to C ₆ alkynyl, each of which is optionally substituted; Optional substituents on R ₄ are one or more substituents independently selected from: amino, mono- (optionally substituted) C ₁ to C ₆ alkylamino, di- (optionally substituted) C ₁ to C ₆ alkylamino, Tri- (optionally substituted) C ₁ to C ₆ alkylammonio, halogeno, carbonyl, trifluoromethyl, nitro, hydroxyl, formyl, cyano, formylamino, carboxyl, azido, sulfo, already Furnace, carbamoyl, formamido, mono- or di-C ₁ to C ₆ alkylcarbamoyl, carbamoyloxy, mono- or di-C ₁ to C ₆ alkylcarbamoyloxy, optionally substituted C ₁ to C ₆ Alkoxy, optionally substituted C ₁ to C ₆ alkoxycarbonyl, optionally substituted C ₁ to C ₆ alkylcarbonyloxy, optionally substituted C ₁ to C ₆ alkylthio, sulfinyl, mono- or di-C ₁ to C ₆ alkylsulfinyl, sulfonyl, mono- or di-C ₁ to C ₆ alkylsulfonyl, sulfamoyl, mono or di-C ₁ G ₆ C ₆ alkylalkylsulfamoyl, sulfinamoyl, mono- or di-C ₁ to C ₆ alkylsulfamoyl, optionally substituted C ₁ to C ₆ alkylcarbonyl, optionally substituted C ₁ to C ₆ alkylcarbonylamino , Optionally substituted C ₁ to C ₆ alkoxycarbonylamino, optionally substituted phenylcarbonyl, optionally substituted heterocyclylcarbonyl, ureido, mono- or di-C ₁ to C ₆ alkylureido, general formula- N (Ra) -C (Rb) = NRc, wherein Ra, Rb and Rc are each hydrogen or optionally substituted C ₁ to C ₆ alkyl, or two of them together are C ₃ to C ₆ cycloalkyl or heterocycle Can form a reel) amidino group, general formula

Wherein the Rabam, Rb and Rc are as defined above;

Wherein R a, R b, R c and R d are each hydrogen or optionally substituted C ₁ to C ₆ alkyl, or two of them may together form C ₃ to C ₆ cycloalkyl or heterocyclyl Anidinyl groups, optionally substituted phenyl, optionally substituted heterocyclyl, C ₁ to C ₆ alkyl and optionally substituted C ₁ to C ₆ alkyl, wherein the substituents are amino, carboxyl, cyano, sulfo, halogeno, carbo Neyl, trifluoromethyl, nitro, hydroxyl, formyl, mono- (optionally substituted) C ₁ to C ₆ alkylamino, di- (optionally substituted) C ₁ to C ₆ alkylamino, tertiary- (optionally Substituted) C ₁ to C ₆ alkylammonio, carbamoyl, mono- or di-C ₁ to C ₆ alkyl carbamoyl, carbamoyloxy, mono- or di-C ₁ to C ₆ alkyl carbamoyloxy, general formula -N (Ra) -C (Rb) = NRc, wherein Ra, Rb and Rc are each hydrogen or optionally substituted C ₁ to C ₆ alkyl Amidino groups of which two adjacent ones together may form C ₃ to C ₆ cycloalkyl or heterocyclyl), and

Wherein Ra, Rb and Rc are as defined above). The method of claim 1 wherein the reaction is carried out in the presence of a base. 3. The base of claim 2 wherein the base is triethyl amine, diisopropylethylamine, 1,8-diaza-bicyclo [5,4,0] -7-undecene, N-methylmorpholine, sodium hydroxide, hydroxide Potassium, potassium carbonate, sodium carbonate, potassium tert-butoxide, sodium methoxide, sodium amide and sodium hydroxide. The process according to claim 1, wherein a reactive derivative of a thiol of the general formula is used in place of the thiol compound of the general formula (III). MS-R ₃ Wherein M is an alkali metal; R ₃ is as defined in claim 1. The process according to claim 1, wherein the compound of formula (II) is a product obtained by oxidizing a compound of formula (IV).

Wherein R ₁ ,

And R ₅ is as defined in claim 1. The method of claim 5, wherein R ₁ is a 1-hydroxyethyl group. The method of claim 6, wherein the substituent R ₃ is selected from the following compounds.

Wherein R ₄ is as defined in claim 1. The method according to claim 1, wherein R ₃ in general formulas (I) and (III) is selected from compounds of the following general formula.

Wherein A is nitrogen or a carbon atom; R ₄ is as defined in claim 1. The compound of claim 8, wherein R ₄ is methyl, ethyl, butyl, cyclopropylmethyl, fluoroethyl, chloroethyl, methoxymethyl, methoxyethyl, aminoethyl,

How to choose from. The method of claim 1 wherein the penem derivative is a compound of formula (la):

Wherein R ₂ is hydrogen or a carboxylate anion as COOR ₂ ; R ₄ is —CH ₃ , —CH ₂ CN, —CH ₂ —C≡CH, —CH ₂ COCH ₃ , —CH ₂ CONH ₂ or —CH ₂ CH ₃ . The method of claim 1 wherein the penem derivative is a compound of formula (lb):

Wherein R ₂ is hydrogen or a carboxylate anion as COOR ₂ ; R ₄ is methyl. The method of claim 1 wherein the penem derivative is a compound of formula (Ic)

Wherein R ₂ is hydrogen or a carboxylate anion as COOR ₂ ; R ₄ is methyl.