KR100192740B1

KR100192740B1 - Antibiotic c-3 dithioacetal-substituted carbapenem compounds, preparation, compositions and their use

Info

Publication number: KR100192740B1
Application number: KR1019910023047A
Authority: KR
Inventors: 알렝 마르텔; 바샹 캐롤; 다리 장폴
Original assignee: 스티븐 비. 데이비스; 브리스톨-마이어스스퀴브컴파니
Priority date: 1991-12-13
Filing date: 1991-12-13
Publication date: 1999-06-15
Also published as: KR930012766A

Abstract

다음 일반식의 디티오아세탈 카르바페넴 또는 비독성의 약리적으로 허용되는 염, 생리학적으로 가수분해가능한 그의 에스테르 또는 용매화물.Dithioacetal carbapenem or a nontoxic pharmacologically acceptable salt thereof, a physiologically hydrolysable ester or solvate thereof, of the general formula:

상기식에서 R¹은 수소 또는 C_1-6알킬이며; n은 0,1 또는 2이며; R³는 수소 또는 C_1-6알킬이며; R²는 C_1-6알킬, 시아노, -CONH₂, -CH₂OH, -CH₂NH₂, -CONHNH₂또는 할로겐 원자수 5개까지, C_1-6알킬 또는 C_1-6알킬록시기로서 임의치환된 페닐, 페닐환위에 할로겐원자수 5개까지, C_1-6알킬 또는 C_1-6알킬록시기로서 임의치환된 페닐메틸, 또는 다음식으로 표시된 라디칼이며In which R ¹ is hydrogen or C _1-6 alkyl; n is 0, 1 or 2; R ³ is hydrogen or C _1-6 alkyl; R ² is C _1-6 alkyl, cyano, -CONH ₂ , -CH ₂ OH, -CH ₂ NH ₂ , -CONHNH ₂ or up to 5 halogen atoms, C _1-6 alkyl or C _1-6 alkyllock Optionally substituted phenyl, phenylmethyl optionally substituted as C _1-6 alkyl or C _1-6 alkyloxy group with up to 5 halogen atoms on the phenyl ring, or a radical represented by the following formula:

상기식에서 p는 0 또는 1이며; X는 C_1-6알킬기로 임의치환된, 황 1개, 산소 1개 또는 질소원자 4개까지 함유한 5원 방향족 헤테로시클릭환, 또는 C_1-6알킬기로 임의치환된, 질소원자 4개까지 함유한 6원 헤테로시클릭환이다.In which p is 0 or 1; The X is an optionally substituted C _1-6 alkyl group, one sulfur, optionally substituted with a 5-membered aromatic heterocyclic ring, or C _1-6 alkyl group containing up to four nitrogen atoms or one oxygen and four nitrogen atoms 6-membered heterocyclic ring.

Description

항상제 C-3 디티오아세탈-치환 카르바페넴 화합물, 그의 조성물, 제조방법 및 용도Homeopathic C-3 Dithioacetal-Substituted Carbapenem Compounds, Compositions, Methods of Use, and Uses

본 발명은 3-위치에 디티오아세탈 부분을 갖는 신규의 경구투여 가능한 카르바페넴 항생제 및 약리적으로 허용가능한 그의 비독성염에 관한 것이다. 본 발명의 카르바페넴은 항미생물 활성을 갖는 것으로 밝혀졌다. 그러므로, 본 발명의 카르바페넴 항생제 및 그의 약리적 조성물은 사람 및 기타 동물의 항균감염 치료시 단독으로 또는 기타 항생제와 배합 사용될 수 있다.The present invention relates to novel orally administrable carbapenem antibiotics having a dithioacetal moiety in the 3-position and pharmacologically acceptable non-toxic salts thereof. Carbapenems of the invention have been found to have antimicrobial activity. Therefore, the carbapenem antibiotic of the present invention and its pharmacological composition may be used alone or in combination with other antibiotics in the treatment of antimicrobial infections in humans and other animals.

본문에는 상기 카르바페넴 항생제의 제조방법과 어떤 신규의 중간체도 기재되어 있다.The text also describes methods of making such carbapenem antibiotics and any novel intermediates.

이 부류 화합물의 명명은 통칭명을 이용하는 루트명인 카르바페넴에 기초하거나 또는 화학초록(Chemical Abstracts)에 따른 계통명에 기초할 수 있다. 본 발명에서는, 예컨대 다음 식의 화합물을 3-R³-4-R²-6-R¹-7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실산이라 명명하는 바와 같이, 화학초록 시스템에 따라 위치에 번호를 매겼다.Naming of this class of compounds may be based on carbapenem, the root name using the generic name, or based on the systematic name according to Chemical Abstracts. In the present invention, for example, the compound of the following formula is 3-R ^3-4 -R ² -6-R 1-7-oxo- ¹ -azabicyclo [3.2.0] hep-2-ten-2-carboxylic acid. As named, locations were numbered according to the chemical abstract system.

본문에서 화합물의 한 부류로서 카르바페넴이라는 용어를 사용하는 것은 7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실산의 표현과 상호교환 가능하다. 그러나, 모든 경우, 본문에 사용된 넘버링 시스템은 상술한 바와 같이 화학초록에 따른 넘버링 시스템이다.The use of the term carbapenem as a class of compounds in the text is interchangeable with the expression of 7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylic acid. In all cases, however, the numbering system used in the text is a numbering system according to the chemical abstract as described above.

기술분야에는 대단히 많은 카르바페넴 항생제가 알려져 있다. 이 부류의 항생제는 발효에 의해 최초로 단리되어 광범위한 항생제 활성을 나타낸 티에나마이신(미국특허 No. 3,950,357, 1976. 4. 13자 Kahan등)에 의해 활자화된다. 이어서, 화학적으로 보다 안정한 티에나마이신의 유도체인 이미페넴(미국특허 NO. 4,194,047, 1980. 3. 18자, Christensen등)이 개발되었다.A great many carbapenem antibiotics are known in the art. This class of antibiotics is first isolated by fermentation and typeset by thienamycin (US Pat. No. 3,950,357, April 13, 1976, Kahan et al.), Which exhibited extensive antibiotic activity. Subsequently, imipenem (US Patent No. 4,194,047, March 18, 1980, Christensen et al.), A chemically more stable derivative of thienamycin, was developed.

보다 최근에 개발된 것은 Shih등에 기재된 다음 구조식(II)의 3-(치환티오)-4-메틸카르바페넴이다(Heterocycles, 21, 29-40 (1984)).More recently developed is 3- (substitutedthio) -4-methylcarbapenem of the following formula (II) described in Shih et al. (Heterocycles, 21, 29-40 (1984)).

Choung등의 1987. 7. 28자 미국특허 No. 4,683,301은 다음 일반식 (III)의 카르바페넴에 관한 것이다.Choung et al. July 28, 1987, US Patent No. 4,683,301 relates to carbapenem of the following general formula (III).

식중 R¹⁰과 R¹¹은 각각 치환 또는 비치환 알킬, 시클로알킬, 페닐 또는 함께 C_5-6알킬리덴을 구성한다.Wherein R ¹⁰ and R ¹¹ each represent substituted or unsubstituted alkyl, cycloalkyl, phenyl or together form C _5-6 alkylidene.

Dextraze의 1989. 11. 14자 미국특허 No. 4,880,922는 다음 식의 카르바페넴에 관한 것이다.Dextraze, US Patent No. 4,880,922 relates to carbapenems of the formula

식중, R은 수소 또는 알킬이고, R⁶은 C_1-6알킬이며; n은 1 내지 3이고Wherein R is hydrogen or alkyl and R ⁶ is C _1-6 alkyl; n is 1 to 3

은 O, S 또는 N 중에서 선택된 부가적인 헤테로 원자를 0-3개 함유하는 방향족 5-또는 6-원 N-함유 헤테로시클릭 고리이고, 상기 방향족 고리는 고리상의 탄소 또는 질소원자에서 C_1-6치환기에 의해 임의로 치환될 수 있으며 상기 고리는 S를 통해 고리 탄소원자에 결합되어 있고 R⁶기에 의해 4급화된 고리질소를 갖는다.Is an aromatic 5- or 6-membered N-containing heterocyclic ring containing 0-3 additional heteroatoms selected from O, S or N, said aromatic ring being C _1-6 in the carbon or nitrogen atom on the ring; Optionally substituted by a substituent, said ring having a ring nitrogen attached to the ring carbon atom via S and quaternized by a R ⁶ group.

1984. 7. 2.자 공개된 두가지 유럽특허출원 Nos. 169,410 및 168,707에는 여러가지 카르바페넴이 개시되어 있는데, 그중에는 다음의 화합물도 있다.Two European patent applications Nos. Various carbapenems are disclosed in 169,410 and 168,707, including the following compounds.

식중 R은 수소 또는 메틸이다.Wherein R is hydrogen or methyl.

본 발명과 관련된 또다른 종래 기술로 Sato등의 The Journal of Antibiotics, 40, 4, pp. 483-495 (1987)을 들 수 있으며, 여기에는 다음 구조를 갖는 카르바페넴이 개시되어 있다.Another conventional technique related to the present invention is described in Sato et al. The Journal of Antibiotics, 40, 4, pp. 483-495 (1987), which discloses carbapenems having the structure:

이미페넴과 티에나마이신 이래 카르바페넴의 항생제 활성은 안정성과 대상 스펙트럼 면에서 진보되었음에도 불구하고, 이렇다할 경구투여성 및 경구활성을 갖는 카르바페넴은 이제까지 보고된 바 없다. 놀랍게도, 본 발명자들은, 본 발명의 어떤 디티오아세탈 및 디티오케탈 카르바페넴이 상당한 경구투여성과 경구 항생제 활성을 갖는다는 것을 발견하였다. 따라서, 본 발명의 목적은 강력한 생체외 항미생물 활성에 더하여 예기치 않은 특성을 갖는 것으로 밝혀진 신규한 부류의 카르바페넴을 제공하는 것이다. 그러므로, 본 발명의 화합물은 인간 및 기타 동물의 감염성 질환의 치료에 대단히 유용하다.Although the antibiotic activity of carbapenems since imipenem and thienamycin has advanced in terms of stability and target spectrum, carbapenems with oral administration and oral activity have not been reported so far. Surprisingly, the inventors have found that certain dithioacetal and dithioketal carbapenems of the present invention have significant oral antibiotics and oral antibiotic activity. It is therefore an object of the present invention to provide a new class of carbapenems that has been found to have unexpected properties in addition to potent ex vivo antimicrobial activity. Therefore, the compounds of the present invention are very useful for the treatment of infectious diseases in humans and other animals.

본 발명은 다음 일반식(I)을 갖는 신규한 3-디티오아세탈 치환 카르바페넴 항생제를 제공한다.The present invention provides novel 3-dithioacetal substituted carbapenem antibiotics having the following general formula (I).

식중, R¹은 수소 또는 C_1-6알킬; n은 0, 1 또는 2; R³은 수소 또는 C_1-6알킬; R²는 C_1-6알킬.Wherein R ¹ is hydrogen or C _1-6 alkyl; n is 0, 1 or 2; R ³ is hydrogen or C _1-6 alkyl; R ² is C _1-6 alkyl.

시아노, -CONH₂, -CH₂OH, -CH₂NH₂, -CONHNH₂로 임의 치환되거나 또는 5개 이하의 할로겐 원자, C_1-6알킬 또는 C_1-6알킬옥시로 임의 치환된 페닐, 페닐고리상에 5개 이하의 할로겐원자, C_1-6알킬 또는 C_1-6알킬옥시기가 임의 치환된 페닐메틸이거나 또는, 다음 식으로 나타내는 기이다.Phenyl optionally substituted with cyano, -CONH ₂ , -CH ₂ OH, -CH ₂ NH ₂ , -CONHNH ₂ or optionally substituted with up to 5 halogen atoms, C _1-6 alkyl or C _1-6 alkyloxy Or phenylmethyl in which up to 5 halogen atoms, C _1-6 alkyl or C _1-6 alkyloxy group are optionally substituted on the phenyl ring, or a group represented by the following formula.

상기중, p는 0 또는 1; X는 황원자 1개, 산소원자 1개 또는 질소원자 4개까지 함유하고, C_1-6알킬기로 임의 치환된 방향족 5원 헤테로시클릭 고리이거나 또는 질소원자를 4개까지 함유하고, C_1-6알킬기로 임의 치환된 방향족 6원 헤테로시클릭 고리이다.Wherein p is 0 or 1; X is a sulfur atom one, containing up to containing one or four nitrogen atoms and an oxygen atom, or an optionally substituted aromatic 5-membered heterocyclic ring as a C _1-6 alkyl group or a nitrogen atom up to 4, and C _1-6 Aromatic 6-membered heterocyclic ring optionally substituted with an alkyl group.

또다른 측면에서, 본 발명은 일반식(I)의 화합물 및 약리적으로 허용되는 그의 비독성염, 생리적으로 가수분해 가능한 에스테르 또는 용매 화합물에 관한다.In another aspect, the invention relates to compounds of formula (I) and pharmacologically acceptable non-toxic salts, physiologically hydrolysable esters or solvent compounds.

본 발명의 대표적인 카르바페넴을 테스트하기 위해 선정하였으며 이들은 강력한 항미생물 활성 및 예기치 않은 경구 투여성을 나타내었다. 그러므로, 본 발명의 추가적인 측면은 상기 카르바페넴 항생제를 유효성분으로 하는 약리적 조성물 및 상기 카르바페넴 항생제 또는 그의 약리적 조성물을 투여하는 것으로 되는 치료방법에도 관계된다.Representative carbapenems of the invention were selected for testing and they showed potent antimicrobial activity and unexpected oral administration. Therefore, a further aspect of the present invention also relates to a pharmacological composition comprising the carbapenem antibiotic as an active ingredient and a treatment method comprising administering the carbapenem antibiotic or a pharmacological composition thereof.

본 발명은 다음 일반식을 갖는 신규한 3-디티오아세탈 치환 카르바페넴 항생제를 제공한다.The present invention provides novel 3-dithioacetal substituted carbapenem antibiotics having the general formula:

식중, R¹, R², R³및 n은 상기 정의한 바와 같다.Wherein R ¹ , R ² , R ³ and n are as defined above.

보다 바람직한 일반식(I)의 화합물은 R¹이 수소 또는 메틸이고; n은 0 또는 1; R³은 수소 또는 C_1-6알킬; R²는 시아노, -CONH₂, -CH₂OH, -CH₂NH₂, -CONHNH₂또는 5개 이하의 할로겐 원자 또는 C_1-6알킬기로 임의 치환된 페닐, 페닐고리상에 5개 이하의 할로겐 원자, 또는 C_1-6알킬기로 임의 치환된 페닐메틸, C_1-6알킬 또는 다음 식으로 나타내는 기이다.More preferred compounds of formula (I) are those in which R ¹ is hydrogen or methyl; n is 0 or 1; R ³ is hydrogen or C _1-6 alkyl; R ² is no more than 5 on cyano, -CONH ₂ , -CH ₂ OH, -CH ₂ NH ₂ , -CONHNH ₂ or phenyl optionally substituted with up to 5 halogen atoms or C _1-6 alkyl groups a halogen atom, or an optionally substituted phenyl C _1-6 alkyl group, methyl, a group represented by C _1-6 alkyl or the following formula:

상기중 p는 0 또는 1; X는 피리딜, 퓨릴 또는 다음 식을 갖는 래디칼이다.P is 0 or 1; X is pyridyl, furyl or a radical having the formula

상기중 Y는 황 또는 산소이다.Wherein Y is sulfur or oxygen.

본 발명은 또한 유용한 중간체 및 일반식(I)의 화합물과 그의 중간체의 제조방법도 제공한다.The present invention also provides useful intermediates and compounds of formula (I) and methods of preparing the intermediates thereof.

일반식(I) 화합물의 상기 정의에서, C_1-6알킬이란 메틸, 에틸, n-프로필, 이소프로필, n-부틸, n-펜틸, n-헥실, 3-메틸펜틸, 등의 알킬과 같은 직쇄 또는 가지달린 알킬을 말하고; 5개 이하의 할로겐 원자 또는 C_1-6알킬기로 치환된 페닐은 2,3,4,5,6-헥사플루오로페닐, 2-플루오로-3-메틸페닐, 2-에틸-3-메틸페닐, 4-메틸페닐, 4-브로모-3-클로로-5-메틸페닐, 4-t-부틸페닐, 3,5-디클로로페닐, 등의 기를 가리키며, 황원자 1개, 산소원자 1개, 또는 질소원자 4개를 함유하는 방향족 5원 헤테로시클릭기는 티에닐, 퓨릴, 피롤릴, 이미다졸릴, 피라졸릴, 티아졸릴, 이소티아졸릴, 옥사졸릴, 이속사졸릴, 트리아졸릴, 티아디아졸릴, 옥사디아졸릴, 테트라졸릴, 티아트리아졸릴, 옥사트리아졸릴 등의 고리를 의미하며, 4개 이하의 질소원자를 함유하는 방향족 6원 헤테로시클릭 고리는 피리딜, 피리미딜, 피라지닐, 피리다지닐, 트리아지닐, 테트라지닐, 등의 방향족 고리를 의미하고; 할로겐원 요오도, 클로로, 플루오로 또는 브로모를 의미한다.In the above definition of the compound of general formula (I), C _1-6 alkyl means an alkyl such as methyl, ethyl, n-propyl, isopropyl, n-butyl, n-pentyl, n-hexyl, 3-methylpentyl, etc. Straight chain or branched alkyl; Phenyl substituted with up to 5 halogen atoms or C _1-6 alkyl groups is 2,3,4,5,6-hexafluorophenyl, 2-fluoro-3-methylphenyl, 2-ethyl-3-methylphenyl, 4 -Methylphenyl, 4-bromo-3-chloro-5-methylphenyl, 4-t-butylphenyl, 3,5-dichlorophenyl, etc., and one sulfur atom, one oxygen atom, or four nitrogen atoms Aromatic five-membered heterocyclic groups containing thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxdiazolyl, tetra A ring such as zolyl, thiatriazolyl, oxatriazolyl, etc., and aromatic six-membered heterocyclic ring containing up to 4 nitrogen atoms include pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, tetra An aromatic ring such as genyl; Meaning halogen source iodo, chloro, fluoro or bromo.

약리적으로 허용되는 비독성염이라함은 비독성산 및 염기성염, 그리고 쯔비터이온 종류의 염을 의미한다. 염기와의 염에는 나트륨, 칼륨, 칼슘 및 마그네슘, 암모늄염과 같은 무기 금속염, 및 트리알킬아민, 피리딘, 피콜린, 디벤질아민, 에탄올아민, N-메틸모르폴린 및 카르복실산과 염을 형성하는데 사용되는 기타 아민과 같은 비독성 아민과의 염이 포함된다. 산과의 염에는 히드로클로라이드, 히드로브로마이드, 히드로요오다이드, 설페이트, 포스페이트 등과 같은 무기산염, 염기성 아민과 염을 형성하는데 사용되는 포르메이트, 아세테이트, 말리에이트, 시트레이트, 숙시네이트, 아스코르베이트, 락테이트, 푸마레이트 및 타르트레이트가 포함된다.Pharmacologically acceptable non-toxic salts refer to non-toxic acid and basic salts and salts of the zwitterionic species. Salts with bases are used to form salts with inorganic metal salts such as sodium, potassium, calcium and magnesium, ammonium salts, and trialkylamines, pyridine, picoline, dibenzylamine, ethanolamine, N-methylmorpholine and carboxylic acids. Salts with non-toxic amines such as other amines. Salts with acids include inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate and the like, formate, acetate, maleate, citrate, succinate, ascorbate, used to form salts with basic amines, Lactate, fumarate and tartrate.

생리적으로 가수분해가능한 에스테르는 체내에서 가수분해되어 항생물질 자체를 발생케하는 전구약물(prodrug)로서 작용한다. 많은 경우 가수분해는 주로 소화효소의 영향하에 일어나므로 이들은 경구투여되는 것이 바람직하다. 에스테르 자체가 활성적이거나 또는, 가수분해가 혈액내에서 일어나는 경우에는 비경구투여를 이용할 수 있다. 일반식(I)의 생리적으로 가수분해가능한 에스테르의 예에는 C_1-6알킬, 벤질, 4-메톡시벤질, 인다닐, 프탈리딜, C_1-6알카노일옥시(C_1-6)알킬, 예컨대, 아세톡시메틸, 피발로일옥시메틸 또는 프로피오닐옥시메틸, C_1-6알콕시카르보닐옥시(C_1-6)알킬, 예컨대 메톡시카르보닐옥시메틸 또는 에톡시카르보닐옥시메틸, 글리실옥시메틸, 페닐글리실옥시메틸, (5-메틸-2-옥소-1,3-디옥솔렌-4-일)메틸 및 페닐실린과 세팔로스포린 기술분야에서 사용되는 공지의 기타 생리적으로 가수분해가능한 에스테르를 들 수 있다.Physiologically hydrolysable esters act as prodrugs that hydrolyze in the body to give rise to the antibiotic itself. In many cases hydrolysis occurs primarily under the influence of digestive enzymes, so they are preferably administered orally. Parenteral administration may be used if the ester itself is active or if hydrolysis occurs in the blood. Examples of physiologically hydrolyzable esters of general formula (I) include C _1-6 alkyl, benzyl, 4-methoxybenzyl, indanyl, phthalidyl, C _1-6 alkanoyloxy (C _1-6 ) alkyl For example, acetoxymethyl, pivaloyloxymethyl or propionyloxymethyl, C _1-6 alkoxycarbonyloxy (C _1-6 ) alkyl, such as methoxycarbonyloxymethyl or ethoxycarbonyloxymethyl, article Lysyloxymethyl, phenylglycosyloxymethyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl and other physiologically hydrolysates known in the art of phenylsilin and cephalosporin Possible esters are mentioned.

본 발명의 화합물은 몇개의 비대칭 탄소원자를 가지며 따라서 몇가지 입체화학 형태로 존재할 수 있다. 본 발명은 이성체의 혼합물과 각 입체이성체를 포함한다. 가장 바람직한 일반식(I)의 화합물은 1-아자비시클로 [3.2.0] 헵탄 고리 구조상에 4R, 5S, 6S 배치를 갖는 것들이다. 덧붙여, 3-치환체는 비대칭 탄소원자 및/또는 R이나 S배치로 존재하는 술피닐기를 함유할 수 있다. 본 발명에는 예컨대, 3-[[[(p-클로로페닐)술피닐]메틸]티오], 3-[[[[(피리디-3-닐)메틸]술피닐]메틸]티오], 3-[[(메틸술피닐)메틸]티오] 및 3-[[1-(메틸티오)에틸]티오]치환체의 R 및 S 이성체와 같이 3-치환체의 R 및 S 이성체가 모두 포함된다.The compounds of the present invention have several asymmetric carbon atoms and can therefore exist in several stereochemical forms. The present invention includes mixtures of isomers and each stereoisomer. Most preferred compounds of formula (I) are those having the 4R, 5S, 6S configuration on the 1-azabicyclo [3.2.0] heptane ring structure. In addition, the 3-substituent may contain asymmetric carbon atoms and / or sulfinyl groups present in the R or S configuration. In the present invention, for example, 3-[[[(p-chlorophenyl) sulfinyl] methyl] thio], 3-[[[[(pyridin-3-yl) methyl] sulfinyl] methyl] thio], 3- Both R and S isomers of the 3-substituted are included, such as the R and S isomers of the [[(methylsulfinyl) methyl] thio] and 3-[[1- (methylthio) ethyl] thio] substituents.

일반식(I)의 신규 카르바페넴 유도체 또는 약리적으로 허용되는 그의 염은 여러가지 그램-양성 및 그램-음성 세균에 대해 활성적인 강력한 항생제이며, 예컨대, 발육촉진을 위한 사료첨가물, 식품에 사용되는 방부제, 산업상 이용시, 예컨대 수성 페인트 및 유해 세균의 생장을 저지하기 위한 페이퍼 밀의 화이트 워터에서의 살균제 및 의료 및 치과용 장비상에 유해 세균이 증식하는 것을 저해 또는 박멸하기 위한 소독제 등으로서 사용될 수 있다. 그러나, 이들은 그램-양성 또는 그램-음성 세균에 기인한 인간 및 기타 동물의 감염성 질환의 치료시 특히 유용하다.Novel carbapenem derivatives of formula (I) or pharmacologically acceptable salts thereof are potent antibiotics active against various gram-positive and gram-negative bacteria, for example feed additives for promoting growth, preservatives used in foods In industrial use, for example, as a disinfectant in white water of paper mills to inhibit the growth of aqueous paints and harmful bacteria and as a disinfectant to inhibit or eradicate the growth of harmful bacteria on medical and dental equipment. However, they are particularly useful in the treatment of infectious diseases in humans and other animals due to Gram-positive or Gram-negative bacteria.

본 발명의 약리적으로 활성인 화합물은 단독으로 사용되거나 또는 카르바페넴 활성성분과 약리적으로 허용가능한 담체 또는 희석제로 구성된 약리적 조성물로서 사용될 수 있다. 본 발명 화합물은 여러 수단에 의해 투여되며; 주요한 수단에는 경구, 국소 또는 비경구(정맥 또는 근육주사) 투여가 포함된다. 약리적 조성물에는 캡슐제, 정제, 분말제 등과 같은 고체형태 또는 용액제, 현탁제 또는 유제와 같은 액체형태가 될 수 있다. 바람직한 투여경로인 주사를 위한 조성물은 앰풀이나 다회투여용 용기에 단위투여 형태로 제조될 수 있으며, 현탁, 안정화 및 분산제와 같은 조성제를 함유시킬 수 있다. 조성물은 즉석 사용가능한 형태 또는 멸균수와 같은 적합한 담체와 함께 투여시 재조성되는 분말형태로 될 수 있다.The pharmacologically active compound of the present invention may be used alone or as a pharmacological composition consisting of a carbapenem active ingredient and a pharmacologically acceptable carrier or diluent. Compounds of the invention are administered by several means; Primary means include oral, topical or parenteral (venous or intramuscular) administration. The pharmacological composition may be in solid form such as capsules, tablets, powders, or the like or in liquid form such as solutions, suspensions or emulsions. Compositions for injection, which are the preferred route of administration, may be prepared in unit dosage form in ampoules or in multidose containers, and may contain compositions such as suspensions, stabilizers and dispersants. The compositions may be in ready-to-use form or in powder form which is reconstituted upon administration with a suitable carrier such as sterile water.

투여량은 사용되는 특정 화합물, 특정 조성물, 투여경로, 숙주의 특성 및 상태 그리고 치료부위 및 치료받는 기관에 따라 크게 달라질 수 있다. 특정의 바람직한 투여량 및 투여경로의 선택은, 치료의사의 재량에 맡긴다. 그러나, 일반적으로, 화합물은 약 5 내지 200 ㎎/㎏일의 양으로 포유류 숙주에게 경구 또는 비경구 투여할 수 있다. 투여는 예컨대 1일 3 내지 4회로 나누어 행한다.Dosages can vary widely depending on the particular compound used, the particular composition, the route of administration, the nature and condition of the host and the site of treatment and the organ being treated. The choice of particular preferred dosage and route of administration is left to the discretion of the therapist. Generally, however, the compound can be administered orally or parenterally to a mammalian host in an amount of about 5 to 200 mg / kg days. Administration is carried out, for example, three to four times a day.

일반식(I)의 카르바페넴 화합물은 다음 반응식 A에 도시한 바와 같이, 일반식(IV)의 중간체를 알킬화제(V)로 알킬화시킴으로써 제조할 수 있다.The carbapenem compound of formula (I) can be prepared by alkylating the intermediate of formula (IV) with an alkylating agent (V), as shown in Scheme A below.

반응식중, R¹, R², R³및 n은 상기 정의한 바와 같다. M⁺은 금속 양이온 또는 수소이고, 바람직한 금속 양이온은 은이다. M⁺가 수소이면, 1 단계는 아민염기 존재하에 수행될 수 있으며; 바람직한 아민염기는 트리(C_1-6)알킬아민, 더욱 바람직한 아민염기는 디이소프로필에틸 아민이다. 일반식(V)의 화합물에서, Q는 요오도, 브로모 또는 클로로와 같은 이탈기를 나타낸다.In the scheme, R ¹ , R ² , R ³ and n are as defined above. M ⁺ is a metal cation or hydrogen, and the preferred metal cation is silver. If M ⁺ is hydrogen, step 1 can be carried out in the presence of an amine base; Preferred amine base groups are tri (C _1-6 ) alkylamines, and more preferred amine base groups are diisopropylethyl amines. In compounds of formula (V), Q represents a leaving group such as iodo, bromo or chloro.

일반식(IV) 및 (VI)에서, R⁷은 통상적인 카르복시 보호기이다. 카르복실산기를 보호 또는 차단하기 위해 본 발명에서 사용될 수 있는 통상적인 카르복시-보호기는 당업자에게 잘 알려져 있으며, 바람직하게는, 예컨대, 온화한 조건하에서 화학 환원제를 이용함에 의한 화학적 또는 효소적 가수분해, 자외선 조사 또는 촉매적 수소첨가에 의하여, 분자의 나머지 부분이 전혀 파괴되지 않도록 하는 방법으로 제거될 수 있는 보호기인 것이 바람직하다. 이러한 쉽게 제거가능한 카르복시-보호기에는 C_1-6알킬, 2-나프틸메틸, 4-피리딜메틸, 펜아실, 아세토닐, 2,2,2-트리클로로에틸, 트리메틸실릴 및 t-부틸디메틸실릴과 같은 실릴, 페닐, 고리치환페닐, 예컨대 4-클로로페닐, 톨릴, 및 t-부틸페닐, 페닐(C_1-6)알킬, 고리치환페닐(C_1-6)알킬, 예컨대 벤질, 4-메톡시벤질, 4-니트로벤질(p-니트로벤질), 2-니트로벤질, 벤즈히드릴 및 트리틸, 메톡시메틸, 2,2,2-트리클로로에톡시카르보닐, 벤질옥시메틸, 예컨대, 아세톡시메틸, 프로피오닐옥시메틸과 같은 C_1-6알카노일옥시(C_1-6)알킬, 비닐 및 알릴과 같은 C_2-6알케닐이 포함된다. 특히 바람직한 카르복시 보호기는 벤질, 4-니트로벤질, 2-니트로벤질, 2,4-디메톡시벤질, 4-메톡시벤질, 알릴 및 치환알릴이다. 기타 적절한 카르복시 보호기가 Theodora W. Greene의 Protective Groups in Organic Synthesis (John Wiley Sons, 1981), 제5장에 기재되어 있다.In formulas (IV) and (VI), R ⁷ is a common carboxy protecting group. Conventional carboxy-protecting groups that can be used in the present invention to protect or block carboxylic acid groups are well known to those skilled in the art and are preferably chemical or enzymatic hydrolysis, ultraviolet light, for example by using chemical reducing agents under mild conditions It is preferred to be a protecting group that can be removed by irradiation or catalytic hydrogenation in such a way that the rest of the molecule is not destroyed at all. Such easily removable carboxy-protecting groups include C _1-6 alkyl, 2-naphthylmethyl, 4-pyridylmethyl, phenacyl, acetonyl, 2,2,2-trichloroethyl, trimethylsilyl and t-butyldimethylsilyl Silyl, phenyl, cyclic substituted phenyls such as 4-chlorophenyl, tolyl, and t-butylphenyl, phenyl (C _1-6 ) alkyl, cyclic substituted phenyl (C _1-6 ) alkyl such as benzyl, 4-meth Oxybenzyl, 4-nitrobenzyl (p-nitrobenzyl), 2-nitrobenzyl, benzhydryl and trityl, methoxymethyl, 2,2,2-trichloroethoxycarbonyl, benzyloxymethyl such as ace C _1-6 alkanoyloxy (C _1-6 ) alkyl such as methoxymethyl, propionyloxymethyl, C _2-6 alkenyl such as vinyl and allyl. Particularly preferred carboxy protecting groups are benzyl, 4-nitrobenzyl, 2-nitrobenzyl, 2,4-dimethoxybenzyl, 4-methoxybenzyl, allyl and substituted allyl. Other suitable carboxy protecting groups are described in Protective Groups in Organic Synthesis (John Wiley Sons, 1981), Chapter 5 of Theodora W. Greene.

식(V) 및 (VI)에서, R⁸은 C_1-6알킬; 5개 이하의 할로겐 원자, C_1-6알킬옥시 또는 C_1-6알킬기가 페닐기상에 임의 치환된 페닐메틸; 시아노, -CONH₂또는 5개 이하의 할로겐 원자, C_1-6알킬 또는 C_1-6알킬옥시기가 임의 치환된 페닐; 다음 식의 기In formulas (V) and (VI), R ⁸ is C _1-6 alkyl; Phenylmethyl in which up to 5 halogen atoms, C _1-6 alkyloxy or C _1-6 alkyl groups are optionally substituted on the phenyl group; Phenyl in which a cyano, -CONH ₂ or up to 5 halogen atoms, C _1-6 alkyl or C _1-6 alkyloxy group is optionally substituted; Flag of the following formula

식중 B는 -CH₂OH, -CH₂NH₂또는 -CONHNH₂로 전환될 수 있는 기이고; 또는 다음 식의 기이다.Wherein B is a group which can be converted to -CH ₂ OH, -CH ₂ NH ₂ or -CONHNH ₂ ; Or a group of the following formula.

식중 p 및 X는 상기 정의한 바와 같다.Wherein p and X are as defined above.

B기를 소망되는 래디칼로 전환시키는 것은 제2단계에서 카르복시보호기 R⁷제거와 동시에 수행되는 것이 바람직하다. 예컨대, R⁷이 p-니트로벤질 래디칼이면, 촉매적 가수소분해에 의해 제거할 수 있다. 가수소분해 반응중 전환될 수 있는 것들인 -CH₂OH, -CH₂NH₂및 -CONHNH₂에 대한 특히 간편한 전구물질들은 각각 -CH₂OCO₂PNB, -CH₂N₃및 -CONHHNCO₂PNB이다.The conversion of group B to the desired radicals is preferably carried out simultaneously with the removal of the carboxyprotecting group R ⁷ in the second step. For example, if R ⁷ is p-nitrobenzyl radical, it can be removed by catalytic hydrogenolysis. Particularly convenient precursors to -CH ₂ OH, -CH ₂ NH ₂ and -CONHNH ₂ , which can be converted during the hydrolysis reaction, are -CH ₂ OCO ₂ PNB, -CH ₂ N ₃ and -CONHHNCO ₂ PNB, respectively. to be.

식(I') 및 (I)에서, R⁹은 수소 또는 염기성 염의 양이온이다. 예컨대 보호기 R⁷의 제거가 나트륨 또는 칼륨 양이온 존재하에 수행되며, R⁹는 각각 나트륨 또는 칼륨이 된다. 소망되는 경우, 제3단계에서 본 분야의 기술을 잘 개발함으로써, 황원자를 술피닐 또는 술포닐기로 산화시킬 수 있다.In formulas (I ′) and (I), R ⁹ is a cation of hydrogen or a basic salt. For example, removal of the protecting group R ⁷ is carried out in the presence of sodium or potassium cations and R ⁹ becomes sodium or potassium, respectively. If desired, sulfur atoms can be oxidized to sulfinyl or sulfonyl groups by well developing the art in the third step.

반응식 A에서 M⁺가 Ag⁺및 수소인 경우의 일반식(IV)의 중간체는 각각 다음 반응식 B 및 반응식 C에 나타난 반응순서에 의해 제조될 수 있다.Intermediates of formula (IV) in which M ⁺ is Ag ⁺ and hydrogen in Scheme A can be prepared by the reaction sequences shown in Scheme B and Scheme C, respectively.

반응식 B 및 C에서, R¹및 R⁹는 상기 정의한 바와 같다. 화합물(VIII)의 화합물(VIII)로의 전환은 ClPO(OPh)₂및 DIPEA에 의해 간편하게 수행할 수 있다. 이탈기인 래디칼 -OPO(OPh)₂는 DIPEA와 같은 염기의 존재하에 테트라히드로피라-2-닐 메르캅탄 또는 TrSH로 대체된다. 다른 한편, -OPO(OPh)₂래디칼은 LiSH로도 대체된다. 화합물(X) 및 (IX)내의 황-탄소 결합은 AgNO₃에 의해 절단된다.In Schemes B and C, R ¹ and R ⁹ are as defined above. The conversion of compound (VIII) to compound (VIII) can be conveniently carried out by ClPO (OPh) ₂ and DIPEA. The leaving group radical -OPO (OPh) ₂ is replaced by tetrahydropyran-2-yl mercaptan or TrSH in the presence of a base such as DIPEA. On the other hand, -OPO (OPh) ₂ radicals are also replaced by LiSH. Sulfur-carbon bonds in compounds (X) and (IX) are cleaved by AgNO ₃ .

다음의 특정 실시예는 반응식 A, B 및 C에 나타낸 합성단계를 설명하는 것으로, 본 발명에 이에 한정되는 것은 아니다. 개시된 방법은 본 발명에 특정 기재되지는 않았으나 본 발명에 포함되는 화합물을 생산하기 위해 변화를 가할 수 있다. 또한, 당업자라면 본 발명에 변형을 가하여 동일한 화합물을 약간 다른 방식으로 생산할 수 있음도 이해할 것이다.The following specific examples illustrate the synthetic steps shown in Schemes A, B, and C, but are not limited to the present invention. Although the disclosed method is not specifically described in the present invention, changes may be made to produce a compound included in the present invention. Those skilled in the art will also understand that modifications to the present invention can produce the same compounds in slightly different ways.

본문에 사용된 약자는 기술분야에 잘 알려진 것들이다. 그중 몇가지를 다음에 나타내었다.Abbreviations used in the text are well known in the art. Some of them are shown below.

[약칭][Abbreviated name]

특정하지 않는한 모든 온도는 섭씨(C)이다. 핵자기공명(NMR) 스펙트럼 특성은 백만분의 부(ppm)로 표시된 화학이동(δ)대 참고기준으로 테트라메틸실란(TMS)을 이용한 것으로 나타낸다. 양성자 NMR 스펙트럼 데이타에서 여러 이동에 대한 보고된 상대면적은 분자내의 특정 관능기 유형의 수소원자 수에 상당한다. 다중성에 관한 화학 이동 특성은 넓은 단일(bs), 넓은 이중(bd), 넓은 삼중(bt), 넓은 사중(bq), 단일(s), 다중(m), 이중(d), 사중(q), 삼중(t), 이중의 이중(dd), 삼중의 이중(dt), 및 사중의 이중(dq)로 나타낸다. NMR 스펙트럼에 사용된 용매는 DMSO-d₆(퍼듀테로디메틸술폭시드), D₂O(중수소화수), CDCl₃(듀테로클로로포름) 및 기타 통상적인 중수소화 용매이다. 적외선(IR) 스펙트럼 설명은 관능기 동정치를 갖는 흡수파수(㎝^-1)만을 포함한다. Celite는 Johns-Manville Products Corporation의 규조토의 상표명이다.Unless otherwise specified, all temperatures are in degrees Celsius (C). Nuclear magnetic resonance (NMR) spectral characteristics are indicated using tetramethylsilane (TMS) as a reference for chemical shifts (δ) versus parts per million (ppm). The reported relative area for the various shifts in proton NMR spectral data corresponds to the number of hydrogen atoms of a particular functional group type in the molecule. The chemical transfer properties for multiplicity are broad single (bs), wide double (bd), wide triple (bt), wide quadruple (bq), single (s), multi (m), double (d), and quadruple (q). , Triplet (t), doublet (dd), doublet (dt), and triplet (dq). Solvents used in the NMR spectrum are DMSO-d ₆ (perduterodimethylsulfoxide), D ₂ O (deuterated water), CDCl ₃ (deuterochloroform) and other conventional deuterated solvents. Infrared (IR) spectral descriptions include only the absorption frequency (cm ⁻¹ ) with functional group identification. Celite is a trademark of diatomaceous earth of Johns-Manville Products Corporation.

[실시예 1]Example 1

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo [3.2.0] hept -2-ten-2-carboxylate

[방법 I][Method I]

[A 단계][Step A]

p-니트로벤질 (4R,5S,6S)-6- [1'(R)히드록시에틸] -4-메틸-3- [(테트라히드로피라-2-닐)티오]-7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) hydroxyethyl] -4-methyl-3-[(tetrahydropyran-2-yl) thio] -7-oxo-1- Azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

건조 CH₃CN(300 ㎖)중의 p-니트로벤질(2R,4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3,7-디옥소-1-아자비시클로 [3.2.0] 헵탄-2-카르복실레이트 (41.33 g, 114.2 m㏖)의 냉용액(얼음-MeOH 배쓰)을 ClPO(0Ф)₂(27.4 ㎖, 131 m㏖) 및 DIPEA (24.0 ㎖, 137 m㏖)로 적가 처리하였다. 이 혼합물을 30분간 교반하여 에놀 포스페이트를 생성한다음, 이를 테트라히드로피라-2-닐 메르캅탄(15 ㎖, 137 m㏖) 및 DIPEA(25 ㎖, 143 m㏖)로 적가처리하였다. 다음 이 혼합물을 1.5시간(-5℃-0℃) 교반하고 다시 티올(3.7 ㎖, 34 m㏖) 및 DIPEA (6 ㎖, 34 m㏖)로 처리한 후, -20℃에서 18시간 정치하였다. 이를 빙냉 H₂O (600 ㎖)로 희석하고 EtOAc (3 × 400 ㎖)로 추출하였다. 에틸 아세테이트 추출물을 결합시켜 1N HCl (400 ㎖), 냉 H₂O (400 ㎖), 냉 1M 수성 NaHCO₃(400 ㎖), 냉염수(400 ㎖)로 세척후 건조시켜(MgSO₄), 중성의 활성목탄으로 처리하였다. 용매증발후 얻어진 잔사를 EtOAc (70 ㎖) 및 석유에테르(750 ㎖)로 희석하여 표제물질을 침전시켰다. 이 공정을 2회 반복하여 순수한 소망물질(54.3 g, 100%)을 얻었다.P-nitrobenzyl (2R, 4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3,7-dioxo-1- in dry CH ₃ CN (300 mL) A cold solution of azabicyclo [3.2.0] heptane-2-carboxylate (41.33 g, 114.2 mmol) (ice-MeOH bath) was dissolved in ClPO (0Ф) ₂ (27.4 mL, 131 mmol) and DIPEA (24.0 mL). , 137 mmol) was added dropwise. The mixture was stirred for 30 minutes to produce enol phosphate, which was then added dropwise with tetrahydropyra-2-yl mercaptan (15 mL, 137 mmol) and DIPEA (25 mL, 143 mmol). The mixture was then stirred for 1.5 hours (-5 ° C.-0 ° C.), treated again with thiol (3.7 mL, 34 mmol) and DIPEA (6 mL, 34 mmol), and then left at -20 ° C. for 18 hours. It was diluted with ice cold H ₂ O (600 mL) and extracted with EtOAc (3 × 400 mL). Combine ethyl acetate extracts, wash with 1N HCl (400 mL), cold H ₂ O (400 mL), cold 1M aqueous NaHCO ₃ (400 mL), cold brine (400 mL), and dry (MgSO ₄ ). Treated with activated charcoal. The residue obtained after evaporation of the solvent was diluted with EtOAc (70 mL) and petroleum ether (750 mL) to precipitate the title material. This process was repeated twice to give the pure desired material (54.3 g, 100%).

[B 단계][Step B]

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸] -4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo [3.2.0] hept -2-ten-2-carboxylate

건조 CH₃CN (250 ㎖)중 p-니트로벤질(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3- [(테트라히드로피라-2-닐)티오] -7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트(52.7 g, 114 m㏖)의 냉용액을 피리딘 (13.8 ㎖, 171 m㏖) 및 MeOH(250 ㎖)중의 AgNO₃(29.3 g, 171 m㏖)의 냉용액으로 처리하였다. 얻어진 혼합물을 30분간 교반한다음 빙수(50㎖)에 넣고 격렬히 교반하였다. 고체를 여과하고 냉 H₂O (500 ㎖)로 분쇄하였다. 이 고체를 다시 여과수집하고 냉수(500 ㎖), 에테르(2 × 500 ㎖)로 세척후 건조시켜 표제물질을 고체(55 g, 99.5%)로서 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(tetrahydropyran-2-yl) in dry CH ₃ CN (250 mL) ) Thio] -7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (52.7 g, 114 mmol) was dissolved in pyridine (13.8 mL, 171 mmol) and Treated with cold solution of AgNO ₃ (29.3 g, 171 mmol) in MeOH (250 mL). The resulting mixture was stirred for 30 minutes and then poured into ice water (50 mL) and stirred vigorously. The solid was filtered off and triturated with cold H ₂ O (500 mL). The solid was collected again by filtration, washed with cold water (500 mL), ether (2 x 500 mL) and dried to give the title material as a solid (55 g, 99.5%).

[방법 II][Method II]

[A 단계][Step A]

p-니트로벤질(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3- [(트리페닐메틸)티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(triphenylmethyl) thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (50 ㎖)중 p-니트로벤질(4R,5S,6S)-3-디페녹소포스포러스옥시-6- [1'(R)-히드록시에틸] -4-메틸-7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트(4.8 g, 9.6 m㏖)의 냉용액(아이스 배쓰)을 TrSH(7.96 g, 28.8 m㏖) 및 DIPEA (5.1 ㎖, 29.0 m㏖)로 적가처리하고 30분간 교반하였다. 아이스 배쓰를 제거하고 혼합물을 48시간 교반한 후 빙수(50 ㎖)로 희석하였다. 수성 혼합물을 1/1 EtOAc/에테르(4 × 20 ㎖) 혼합물로 추출하였다. 유기층을 결합하고, 냉수(3 × 50 ㎖)로 세척후 건조시켰다(MgSO₄). 실리카겔(실리카 125 g; 먼저 CH₂Cl₂로, 이어서 5%, 10%, 및 15% EtOAc/CH₂Cl₂로 용출) 패드를 통해 잔사를 통과시켜 표제 물질을 얻었다(1.15 g, 19%).P-nitrobenzyl (4R, 5S, 6S) -3-diphenoxyphosphoroxyoxy-6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1- in DMF (50 mL) A cold solution (ice bath) of azabicyclo [3.2.0] hep-2-ten-2-carboxylate (4.8 g, 9.6 mmol) was purified by TrSH (7.96 g, 28.8 mmol) and DIPEA (5.1 mL, 29.0). mmol), and the mixture was stirred for 30 minutes. The ice bath was removed and the mixture was stirred for 48 hours and then diluted with iced water (50 mL). The aqueous mixture was extracted with a 1/1 EtOAc / ether (4 × 20 mL) mixture. The organic layers were combined, washed with cold water (3 × 50 mL) and dried (MgSO ₄ ). Silica gel (silica 125 g; a first CH ₂ Cl _2, then 5%, 10%, and 15% eluted with EtOAc / CH _₂ Cl ₂₎ to through the pad passes through the residue to give the title material (1.15 g, 19%) .

[B 단계][Step B]

p-니트로벤질 (4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo [3.2.0] hept -2-ten-2-carboxylate

무수 CH₃CN (5 ㎖)중 p-니트로벤질(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3- [(트리페닐메틸)티오] -7-옥소-1-아자비시클로 [3.2.0] 헵-2-텐-2-카르복실레이트(842 ㎎, 1.36 m㏖)의 냉(아이스 배쓰) 용액을 피리딘(0.16 ㎖, 2.03 m㏖) 및 MeOH (5 ㎖)중의 AgNO₃(345 ㎎, 2.03 m㏖)로 적가처리하였다. 첨가 후, 혼합물을 5℃에서 1.5시간 교반한 다음 냉수(10 ㎖)로 희석하고 5분간 더 교반하였다. 고체를 여과수집하고, 냉수(2 × 20 ㎖), 에테르(2 × 20 ㎖)로 세척후 고압하에 건조시켜 담갈색 고체로서 표제물질을 얻었다(604 ㎎, 92%).P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(triphenylmethyl) thio]-in anhydrous CH ₃ CN (5 mL)- A cold (ice bath) solution of 7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (842 mg, 1.36 mmol) was pyridine (0.16 mL, 2.03 mmol) and Treated dropwise with AgNO ₃ (345 mg, 2.03 mmol) in MeOH (5 mL). After addition, the mixture was stirred at 5 ° C. for 1.5 hours, then diluted with cold water (10 mL) and stirred for another 5 minutes. The solid was collected by filtration, washed with cold water (2 × 20 mL), ether (2 × 20 mL) and dried under high pressure to give the title material as a light brown solid (604 mg, 92%).

[실시예 2]Example 2

알릴(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] hep-2 -Ten-2-carboxylate

[A 단계][Step A]

알릴(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3- [(테트라히드로피라-2-닐)티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(tetrahydropyra-2-yl) thio] -7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate

CH₃CN (200 ㎖)중 알릴(2R,4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3,7-디옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (13.6 g, 50.8 m㏖)의 냉용액(아이스-MeOH 배쓰)을 ClPO (OФ)₂(11.8 ㎖, 57.1 m㏖), DIPEA (10.3 ㎖, 58.7 m㏖) 및 흔적량의 DMAP (15 ㎎)으로 적가 처리한 후 -10℃-0℃에서 1시간 교반하였다. 얻어진 에놀 포스페이트 용액을 아르곤가스(15분)으로 정화시키고 CH₃CN(25 ㎖)중의 테트라히드로피라-2-닐 메르캅탄(6.6 g, 56 m㏖) 및 DIPEA (10.7 ㎖, 61.0 m㏖)으로 연속 적가 처리하였다. 혼합물을 5℃에서 1시간, 약 22℃에서 20시간 교반하였다. 혼합물을 아이스 배쓰로 냉각하고, CH₃CN(5 ㎖)중 티올(1.5 g, 12.7 m㏖) 및 DIPEA (2.28 ㎖, 13.0 m㏖)로 부가 처리후, 22℃에서 8시간, 5℃에서 18시간 교반시켰다. 혼합물을 빙냉수(250 ㎖)로 희석후 EtOAc (3 × 200 ㎖)로 희석하였다. 에틸아세테이트 추출물을 결합하고 차가운 0.5 N 수성 HCl (1 × 200 ㎖), 물(1 × 200 ㎖), 0.5 M 수성 NaHCO₃(1 × 200 ㎖), 물 (1 × 200 ㎖), 염수(200 ㎖)로 세척후 건조시켰다(MgSO₄). 실리카겔 패드를 통해 잔사를 통과시켜(실리카 200 g; 먼저 CH₂Cl₂, 이어서 2%, 5%, 10% 및 25% EtOAc/CH₂Cl₂로 용출), 약간의 에톨포스페이트로 오염된 표제물질(13.9 g, 75%)을 입체이성체의 혼합물로서 얻었다.Allyl (2R, 4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3,7-dioxo-1-azabicyclo- [in CH ₃ CN (200 mL) [ 3.2.0] Hept-2-ten-2-carboxylate (13.6 g, 50.8 mmol) in cold solution (Ice-MeOH bath) with ClPO (OФ) ₂ (11.8 mL, 57.1 mmol), DIPEA (10.3 ML, 58.7 mmol) and trace amount of DMAP (15 mg) were added dropwise, followed by stirring at −10 ° C.-0 ° C. for 1 hour. The resulting enol phosphate solution was clarified with argon gas (15 minutes) and then with tetrahydropyran-2-yl mercaptan (6.6 g, 56 mmol) and DIPEA (10.7 mL, 61.0 mmol) in CH ₃ CN (25 mL). Continuous dropwise treatment. The mixture was stirred at 5 ° C. for 1 hour and at about 22 ° C. for 20 hours. The mixture was cooled in an ice bath and after addition treatment with thiol (1.5 g, 12.7 mmol) and DIPEA (2.28 mL, 13.0 mmol) in CH ₃ CN (5 mL), 8 h at 22 ° C., 18 at 5 ° C. Stirred for time. The mixture was diluted with ice cold water (250 mL) and then diluted with EtOAc (3 x 200 mL). Combine ethyl acetate extract and cold 0.5 N aqueous HCl (1 × 200 mL), water (1 × 200 mL), 0.5 M aqueous NaHCO ₃ (1 × 200 mL), water (1 × 200 mL), brine (200 mL ) And then dried (MgSO ₄ ). The residue was passed through a pad of silica gel (200 g of silica; first eluted with CH ₂ Cl ₂ , then 2%, 5%, 10% and 25% EtOAc / CH ₂ Cl ₂ ) to contaminate the title material with some etholphosphate. (13.9 g, 75%) was obtained as a mixture of stereoisomers.

[B 단계][Step B]

H₂O (9 ㎖)중 AgNO₃(442 ㎎, 2.6 m㏖) 용액을 N₂로 10분간 정화시키고 이어서 에테르(4 ㎖)중 피리딘(0.08 ㎖, 1.0 m㏖)을 첨가하였다. 격렬히 교반된 혼합물에 에테르(3 ㎖)중 알릴(4R,5S,6S)-6- [1'(R)-히드록시에틸] -4-메틸-3- [(테트라히드로피라-2-닐)티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (190 ㎎, 0.52 m㏖) 용액을 첨가하고, 암실에서 30분간 더 교반하였다. 침전물을 여과 수집하고, 물(10 ㎖), 에테르(2 × 10 ㎖)로 세척후 건조시켜 표제물질(186 ㎎, 92%)을 갈색고체로 얻었다.A solution of AgNO ₃ (442 mg, 2.6 mmol) in H ₂ O (9 mL) was clarified with N ₂ for 10 min and then pyridine (0.08 mL, 1.0 mmol) in ether (4 mL) was added. To the vigorously stirred mixture was allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(tetrahydropyran-2-yl) in ether (3 mL). A thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (190 mg, 0.52 mmol) solution was added and stirred in the dark for 30 minutes. The precipitate was collected by filtration, washed with water (10 mL), ether (2 × 10 mL) and dried to afford the titled substance (186 mg, 92%) as a brown solid.

[실시예 3]Example 3

p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3-실버 메르캅토-7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] hep-2-ten- 2-carboxylate

[A 단계][Step A]

p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3- [(트리페닐메틸)티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[(triphenylmethyl) thio] -7-oxo-1-azabicyclo- [3.2.0] hepp -2-ten-2-carboxylate

CH₃CN (40 ㎖)중 p-니트로벤질(2R,5S,6S)-6- [1'(R)-히드록시에틸]-3,7-디옥소-1-아자비시클로- [3.2.0] 헵탄-2-카르복실레이트(3.48 g, 10 m㏖)의 냉용액(아이스 배쓰)를 먼저 ClPO(OФ)₂(2.33 ㎖, 11.0 m㏖), 이어서 DIPEA (1.96 ㎖, 11 m㏖)로 처리하여 1시간 교반하였다. 얻어진 에놀 포스페이트 용액에 TrSH (8.3 g, 30 m㏖), 이어서 DIPEA (5.3 ㎖, 30 m㏖)를 적가하였다. 얻어진 혼합물을 약 22℃에서 21시간 교반하고 EtOAc (300 ㎖)로 희석후 H₂O (4 × 200 ㎖), 염수로 세척후 건조시켰다(MgSO₄). 다음 잔사를 실리카겔 패드를 통해 통과시켜(실리카겔 125 g, CH₂Cl₂/EtOAc로 용출) 표제물질(4.87 g, 80%)을 얻었다.P-nitrobenzyl (2R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo- [3.2.0 in CH ₃ CN (40 mL) ] A cold solution of heptane-2-carboxylate (3.48 g, 10 mmol) (ice bath) was first added with ClPO (OФ) ₂ (2.33 mL, 11.0 mmol) followed by DIPEA (1.96 mL, 11 mmol). The mixture was stirred for 1 hour. To the obtained enol phosphate solution was added dropwise TrSH (8.3 g, 30 mmol) followed by DIPEA (5.3 mL, 30 mmol). The resulting mixture was stirred at about 22 ° C. for 21 hours, diluted with EtOAc (300 mL), washed with H ₂ O (4 × 200 mL), brine and dried (MgSO ₄ ). The residue was then passed through a pad of silica gel (125 g of silica gel, eluted with CH ₂ Cl ₂ / EtOAc) to give the title material (4.87 g, 80%).

[B 단계][Step B]

CH₃CN (1 ㎖)중 p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3- [(트리페닐메틸)티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (225 ㎎, 0.37 m㏖)의 냉용액(아이스 배쓰)을 피리딘(34㎕, 0.41 m㏖0 및 MeOH(2.7 ㎖, 0.41 m㏖)중 AgNO₃0.15 M 용액으로 처리후 25분간 교반하였다. 이 혼합물을 빙냉수(5 ㎖)로 희석하고, 침전물을 여과 제거하였다. 고체를 H₂O (1 × 5 ㎖), 에테르(4 × 10 ㎖)로 세척후 고압하에 18시간동안 건조시켜 표제물질(155 ㎎, 89%)을 황색 고체로 얻었다.P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[(triphenylmethyl) thio] -7-oxo-1-azabi in CH ₃ CN (1 mL) A cold solution (ice bath) of cyclo- [3.2.0] hep-2-ten-2-carboxylate (225 mg, 0.37 mmol) was diluted with pyridine (34 μl, 0.41 mmol0 and MeOH (2.7 mL, 0.41). After treatment with 0.15 M solution of AgNO _{3 in} mmol), the mixture was stirred for 25 minutes, the mixture was diluted with ice-cold water (5 mL) and the precipitate was filtered off.The solid was H ₂ O (1 × 5 mL), ether ( 4 x 10 mL) and dried under high pressure for 18 hours to give the titled material (155 mg, 89%) as a yellow solid.

[실시예 4]Example 4

포타슘 또는 소듐 (5R,6S)-6- [1'(R)-히드록시에틸] -3- [[(메틸티오)메틸] 티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (Ia)Potassium or sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ia)

[A 단계][Step A]

p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3- [[(메틸티오)메틸]티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.0 Hep-2-ten-2-carboxylate

p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3-실버 메르캅토-7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (422 ㎎, 0.900 m㏖)의 아세토니트릴(10 ㎖) 용액을 ClCH₂SCH₃(87 ㎕, 0.99 m㏖)로 처리하고 3시간 교반하였다. 다음 혼합물을 진공 농축하고 실리카겔 플래쉬 컬럼에 통과시켜(실리카 20 g, CH₃CN으로 용출) 표제화합물(140 ㎎, 37%)을 얻었다.p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] hep-2-ten- An acetonitrile (10 mL) solution of 2-carboxylate (422 mg, 0.900 mmol) was treated with ClCH ₂ SCH ₃ (87 μl, 0.99 mmol) and stirred for 3 hours. The mixture was then concentrated in vacuo and passed through a silica gel flash column (20 g of silica, eluted with CH ₃ CN) to afford the title compound (140 mg, 37%).

[B 단계][Step B]

포타슘 또는 소듐 (5R,6S)-6- [1'(R)-히드록시에틸] -3- [[(메틸티오] 메틸]티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트 (Ia)Potassium or sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(methylthio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ia)

THF (25 ㎖)중 p-니트로벤질(5R,6S)-6- [1'(R)-히드록시에틸] -3- [(메틸티오)메틸] 티오] -7-옥소-1-아자비시클로- [3.2.0] 헵-2-텐-2-카르복실레이트(300 ㎎, 0.700 m㏖), 에테르(25 ㎖) 및 0.05 M pH 7.0 NaH₂PO₄/NaOH 완충용액(25 ㎖)을 10% Pd/C를 촉매로 하여 45 psi H₂에서 15℃ 내지 20℃, 1시간동안 가수소분해시켰다. 유기층을 pH 7.0 완충액 (2 × 5 ㎖)으로 추출하였다. 수성 분획을 결합하고, 에테르(2 × 20 ㎖) 세척한 후 역상 C₁₈μBondaPak 컬럼에 통과시켜(역상 C₁₈μBondaPak 컬럼물질, 먼저 H₂O, 이어서 2% CH₃CN/H₂O로 용출) 표제화합물(180 ㎎, 78%)을 회백색 고체로 얻었다.P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[(methylthio) methyl] thio] -7-oxo-1-azabicyclo in THF (25 mL) [3.2.0] Hep-2-ten-2-carboxylate (300 mg, 0.700 mmol), ether (25 mL) and 0.05 M pH 7.0 NaH ₂ PO ₄ / NaOH buffer (25 mL) were added. Hydrolysis was performed at 15 ° C. to 20 ° C. for 1 hour at 45 psi H ₂ using% Pd / C as catalyst. The organic layer was extracted with pH 7.0 buffer (2 × 5 mL). Combining the aqueous fraction, and ether (2 × 20 ㎖) After washing and passed through a reverse phase C ₁₈ μBondaPak column (reverse phase C ₁₈ μBondaPak column material, first, H ₂ O, followed by eluting with _{_{2% CH 3 CN / H 2}} O) The title compound (180 mg, 78%) was obtained as an off-white solid.

[실시예 5]Example 5

소듐 또는 포타슘 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)-메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ib)Sodium or potassium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) -methyl] thio] -7-oxo-1-azabi Cyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ib)

[A 단계][Step A]

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabi Cyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (40 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트(4.85 g, 10.0 m㏖)의 냉용액(아이스 배쓰)을 DMF (5 ㎖)중 요오드메틸 메틸 술파이드(1.04 ㎖, 12.5 m㏖) 및 LiI (2.0 g, 15 m㏖)로 서서히 처리하였다. 혼합물을 5℃에서 1시간 교반하고, 빙수(100 ㎖) 및 냉 EtOAc (100 ㎖)로 희석하고 Celite 패드를 통해 여과하였다. 패드를 EtOAc (4 × 100 ㎖)로 세척하고 2 용액상을 분리하였다. 다음 수성상을 EtOAc (2 × 50 ㎖)로 추출하였다. 유기상을 결합시킨다음, 냉수(2 × 150 ㎖), 냉 0.1 N 수성 HCl (150 ㎖), 물(150 ㎖), 0.1M 수성 NaHCO₃(150 ㎖), 물(150 ㎖) 및 염수로 연속 세척후 건조시켰다(MgSO₄). 잔사를 실리카겔(35 g) 패드에 로드시키고 다음의 용매로 연속 용출시킨 후 0℃에서 냉각하였다: 먼저 CH₂Cl₂, 이어서 5, 10 및 20% EtOAc/CH₂Cl₂. 이 정제 단계후, 표제화합물 2.4 g(55%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo in DMF (40 mL) -[3.2.0] Hep-2-ten-2-carboxylate (4.85 g, 10.0 mmol) in cold solution (ice bath) with iodinemethyl methyl sulfide (1.04 mL, 12.5 m) in DMF (5 mL) Mol) and LiI (2.0 g, 15 mmol) slowly. The mixture was stirred at 5 ° C. for 1 h, diluted with iced water (100 mL) and cold EtOAc (100 mL) and filtered through a Celite pad. The pad was washed with EtOAc (4 × 100 mL) and 2 solution phases were separated. The aqueous phase was then extracted with EtOAc (2 × 50 mL). The organic phases are combined and then washed successively with cold water (2 x 150 mL), cold 0.1 N aqueous HCl (150 mL), water (150 mL), 0.1M aqueous NaHCO ₃ (150 mL), water (150 mL) and brine. It was then dried (MgSO ₄ ). The residue was loaded on a pad of silica gel (35 g) and continuously eluted with the following solvent and cooled at 0 ° C .: first CH ₂ Cl ₂ , then 5, 10 and 20% EtOAc / CH ₂ Cl ₂ . After this purification step, 2.4 g (55%) of the title compound were obtained.

[B 단계][Step B]

(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ib')(4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2 .0] hep-2-ten-2-carboxylate (Ib ')

0.1 M NaH₂PO₄/NaOH pH 7.0 완충용액(255 ㎖, 25.5 m㏖), 에테르(70 ㎖) 및 THF (70 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (6.00 g, 13.7 m㏖) 용액을 실온(약 22℃)에서 3.5시간동안 45-50 psi에서 10% Pd/C 촉매(6 g)으로 가수소분해시켰다. 촉매를 여과제거하고 0.1 M 인산염 pH 7.0 완충액(2 × 15 ㎖)로 세척하였다. 그 용액상을 분리하고 유기층을 pH 7.0 완충액(2 × 15 ㎖)으로 추출하였다. 수층을 결합하고, 에테르(3 × 100 ㎖)로 세척후 파티실 역상 컬럼에 통과시켜(파티실 역상물질 275 g, 먼저 H₂O, 이어서 10% CH₃CN/H₂O로 추출) 표제물질(3.37 g, 76%)을 백색고체로서 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 'in 0.1 M NaH ₂ PO ₄ / NaOH pH 7.0 buffer (255 mL, 25.5 mmol), ether (70 mL) and THF (70 mL) (R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxyl The rate (6.00 g, 13.7 mmol) solution was hydrolyzed with 10% Pd / C catalyst (6 g) at 45-50 psi for 3.5 hours at room temperature (about 22 ° C.). The catalyst was filtered off and washed with 0.1 M phosphate pH 7.0 buffer (2 × 15 mL). The solution phase was separated and the organic layer was extracted with pH 7.0 buffer (2 x 15 mL). The aqueous layers were combined, washed with ether (3 × 100 mL) and passed through a patyl reversed phase column (275 g of patyl reversed phase, first extracted with H ₂ O, then 10% CH ₃ CN / H ₂ O). (3.37 g, 76%) was obtained as a white solid.

[A 단계][Step A]

알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [ 3.2.0] hep-2-ten-2-carboxylate

MeOH (1 ㎖)중 알릴(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[(테트라히드로피라-2-닐)티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (104 ㎎, 0.280 m㏖)의 냉용액(아이스-MeOH 배쓰)을 MeOH (3 ㎖)중 AgNO₃(145 ㎎, 0.85 m㏖) 용액 및 피리딘(0.045 ㎖, 0.560 m㏖)로 적가처리하고 -15℃에서 5분간 교반하였다. 형성된 은염을 침전시키기 위해 에테르(15 ㎖)를 첨가하였다. 용매를 따라내고 고체를 에테르(2 × 10 ㎖) 분쇄하였다. 다음 은염을 진공건조하고 CH₃CN (2 ㎖)에 냉각하고 ClCH₂SCH₃(0.10 ㎖, 1.2 m㏖)로 처리하였다. 얻어진 혼합물을 -15℃ 내지 0℃에서 1시간 교반하고, 실리카겔 패드를 통해 통과시켜(실리카 2.5 g, EtOAc로 용출) 농축하여 잔사를 얻고 이를 예비 TLC 플레이트에 적용시켰다(CH₂Cl₂/EtOAc: 1/1). 표제 물질은 오일로서 저수율로 얻어졌다.Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(tetrahydropyran-2-yl) thio] -7- in MeOH (1 mL) A cold solution (ice-MeOH bath) of oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (104 mg, 0.280 mmol) was added to AgNO ₃ (3 mL) in MeOH (3 mL). 145 mg, 0.85 mmol) solution and pyridine (0.045 mL, 0.560 mmol) were added dropwise and stirred at -15 ° C for 5 minutes. Ether (15 mL) was added to precipitate the silver salt formed. The solvent was decanted and the solid was triturated with ether (2 × 10 mL). The silver salt was then dried in vacuo, cooled in CH ₃ CN (2 mL) and treated with ClCH ₂ SCH ₃ (0.10 mL, 1.2 mmol). The resulting mixture was stirred for 1 h at −15 ° C. to 0 ° C., passed through a pad of silica gel (eluted with silica 2.5 g, EtOAc) to give a residue, which was applied to a preparative TLC plate (CH ₂ Cl ₂ / EtOAc: 1/1). The title material was obtained in low yield as an oil.

[B 단계][Step B]

포타슘 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ib)Potassium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [ 3.2.0] hep-2-ten-2-carboxylate (Ib)

알릴(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[(테트라히드로피라-2-닐)티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (2.0 g, 5.45 m㏖)를 상기와 같이 ClCH₂SCH₃으로 처리하여 알릴(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트를 얻고 이를 분리하지 않고 5℃(아이스 배쓰)로 냉각하여 Pd(PФ₃)₄(381 ㎎, 0.33 m㏖), PФ₃(372 ㎎, 1.42 m㏖) 및 EtOAc (20 ㎖)중 에틸헥사노에이트칼륨(3.6 g, 19 m㏖)로 처리하였다. CH₃CN (20 ㎖)를 첨가함으로써, 생성된 침전물을 재용해시키고 용액을 실온(약 22℃)에서 2시간 교반하였다. 혼합물을 에테르(50 ㎖)로 용해하고 0.05 M pH 7.4 인산염 완충액(3 × 10 ㎖)로 추출하였다. 염기성 수성 추출물을 에테르(2 × 25 ㎖)로 세척하고 C₁₈역상 컬럼에 통과시켜(역상물질 80 g; 컬럼은 먼저 H₂O로, 이어서 2%, 5% 및 10% CH₃CN/H₂O로 용출) 불순한 표제 물질을 얻었다. 이 물질을 동결건조시키고 얻어진 분말을 C₁₈역상 컬럼상에서 재정제하여(C₁₈물질 50 g; 컬럼은 먼저 H₂O, 이어서 2% 및 5% CH₃CN/H₂O로 연속 용출) 순수한 표제물질(135 ㎎, 7%)을 얻었다. 이것의 물리적 데이타는 상술한 나트륨염의 것과 동일하다: 순도: 99.94% (HPLC), T¹ _/275시간(pH 7.4)T¹/₂18.8분 (pH 2).Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[(tetrahydropyra-2-yl) thio] -7-oxo-1-azabicyclo -[3.2.0] hep- ₂ -ten-2-carboxylate (2.0 g, 5.45 mmol) was treated with ClCH ₂ SCH ₃ as above to allyl (4R, 5S, 6S) -6- [1 ' (R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxyl Obtained rate and cooled to 5 ° C. (ice bath) without separation, ethyl hexa- Pd (PФ ₃ ) ₄ (381 mg, 0.33 mmol), PФ ₃ (372 mg, 1.42 mmol) and EtOAc (20 mL) Treatment with potassium noate (3.6 g, 19 mmol). By adding CH ₃ CN (20 mL), the resulting precipitate was redissolved and the solution stirred at room temperature (about 22 ° C.) for 2 hours. The mixture was dissolved in ether (50 mL) and extracted with 0.05 M pH 7.4 phosphate buffer (3 × 10 mL). The basic aqueous extract was washed with ether (2 × 25 mL) and passed through a C ₁₈ reverse phase column (80 g of reverse phase; the column first with H ₂ O, then 2%, 5% and 10% CH ₃ CN / H ₂ Eluting with O) the title compound was obtained. The material was lyophilized and the powder obtained was reconditioned on a C ₁₈ reversed phase column (50 g of C ₁₈ material; the column was eluted first with H ₂ O followed by 2% and 5% CH ₃ CN / H ₂ O) pure title The material (135 mg, 7%) was obtained. Its physical data is the same as that described above sodium salt: purity: 99.94% (HPLC), T 1/2 75 sigan ^{_{(pH 7.4) T 1/2}} 18.8 bun (pH 2).

[실시예 6]Example 6

포타슘 또는 소듐 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ic)Potassium or sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ic)

[A 단계][Step A]

p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1- Azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (20 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-실버메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (2.5 g, 5.3 m㏖)의 냉현탁액(아이스 배쓰)를 DMF (5 ㎖)중 4-[(클로로메틸)티오] 피리딘 (1.00 g, 6.36 m㏖)로 적가처리한다음 LiI (1.42 g, 10.6 m㏖) 및 DIPEA (1.1 ㎖, 6.36 m㏖)을 첨가하였다. 약 22℃에서 혼합물을 24시간 교반한다음, H₂O (25 ㎖) 및 EtOAc로 희석후 여과하였다. 2 용액상을 분리하고, 수층을 EtOAc (2 × 40 ㎖)로 추출하였다. 유기상을 결합하고, 빙냉수(5 × 50 ㎖) 및 염수(50 ㎖)로 세척후 건조시켰다(MgSO₄). 용매증발후 회수된 고체잔사를 차가운(5℃) CH₂Cl₂-에테르의 1/1 혼합물(25 ㎖)로 분쇄하여 고체로서 표제물질을 얻고 이를 동일한 용매혼합물(2 × 10 ㎖)로 세척후 건조시켰다(1.47 g). 모액 증발후 얻은 유성잔사를 예비 TLC 플레이트에 적용하여 (EtOAc/CH₃CN: 9/1) 표제물질을 얻었다(총 수율 1.56 g, 60%).P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silvermercapto-7-oxo-1-azabicyclo- [3.2.0] in DMF (20 mL) A cold suspension (ice bath) of hept-2-ten-2-carboxylate (2.5 g, 5.3 mmol) was added with 4-[(chloromethyl) thio] pyridine (1.00 g, 6.36 mmol) in DMF (5 mL). ) Dropwise and LiI (1.42 g, 10.6 mmol) and DIPEA (1.1 mL, 6.36 mmol) were added. The mixture was stirred for 24 h at about 22 ° C., then diluted with H ₂ O (25 mL) and EtOAc and filtered. 2 solution phases were separated and the aqueous layer was extracted with EtOAc (2 × 40 mL). The organic phases were combined, washed with ice cold water (5 × 50 mL) and brine (50 mL) and dried (MgSO ₄ ). The solid residue recovered after evaporation of the solvent was triturated with a 1/1 mixture of cold (5 ° C.) CH ₂ Cl ₂ -ether (25 mL) to obtain the title material as a solid, which was washed with the same solvent mixture (2 × 10 mL). Dried (1.47 g). The oily residue obtained after the mother liquor evaporation was applied to a preparative TLC plate (EtOAc / CH ₃ CN: 9/1) to give the title material (total yield 1.56 g, 60%).

[B 단계][Step B]

포타슘 또는 소듐 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ic)Potassium or sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[((pyridin-4-yl) thio] methyl] thio] -7-oxo-1-azabi Cyclo- [3.2.0] hep-2-ten-2-carboxylate (Ic)

0.05 M pH 7.0 NaH₂PO₄/NaOH 또는 KH₂PO₄/KOH 완충액(40 ㎖, 2.0 m㏖), 에테르(40 ㎖) 및 THF(40 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (750 ㎎, 1.54 m㏖) 용액을 촉매로서 10% Pd/C (750 ㎎)을 이용하여 40 psi H₂에서 1시간 가수소 분해하였다. 촉매를 제거하고 H₂O(5 ㎖)로 세척하였다. 여과물을 에테르(50 ㎖)로 희석하고 유기상은 H₂O(2 × 5㎖)로 추출하였다. 수상을 결합하고 에테르(2 × 50 ㎖)로 세척하여 C₁₈μBondaPak 역상 컬럼에 통과시켜(역상물질 40 g; 먼저 H₂O, 이어서 5%, 10% 및 15% CH₃CN/H₂O로 용출) 표제물질을 얻었다(217 ㎎, 37%); 순도 97.9% (HPLC에 의한 것임); T¹/₂15시간(pH 7.4).P-nitrobenzyl (5R, 6S) -6 in 0.05 M pH 7.0 NaH ₂ PO ₄ / NaOH or KH ₂ PO ₄ / KOH buffer (40 mL, 2.0 mmol), ether (40 mL) and THF (40 mL) -[1 '(R) -hydroxyethyl] -3-[[[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2 -Ten-2-carboxylate (750 mg, 1.54 mmol) solution was hydrolyzed at 40 psi H ₂ for 1 hour using 10% Pd / C (750 mg) as catalyst. The catalyst was removed and washed with H ₂ O (5 mL). The filtrate was diluted with ether (50 mL) and the organic phase was extracted with H ₂ O (2 × 5 mL). The aqueous phases were combined and washed with ether (2 × 50 mL) and passed through a C ₁₈ μBondaPak reversed phase column (40 g of reverse phase; first with H ₂ O followed by 5%, 10% and 15% CH ₃ CN / H ₂ O. Eluting) the title material (217 mg, 37%); Purity 97.9% (by HPLC); T ^1/2 ₂ h (pH 7.4).

[실시예 7]Example 7

포타슘 또는 소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-[[[(페닐티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Id)Potassium or sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-[[[(phenylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (Id)

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(페닐티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(phenylthio) methyl] thio] -7-oxo-1-azabi Cyclo- [3.2.0] hep-2-ten-2-carboxylate

CH₃CN (25 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (760 ㎎, 1.57 m㏖)의 냉용액(아이스 배쓰)를 클로로메틸페닐 술파이드(0.255 ㎖, 1.9 m㏖)로 처리하고 이 혼합물을 30분간 교반하였다. 클로로메틸페닐 술파이드(0.5 ㎖, 3.8 m㏖)을 2회 반복 첨가한 후 각각 30분 및 45분간 교반하였다. 차가운 혼합물을 실리카겔(5 g) 패드에 통과시키고 패드를 차가운 EtOAc (20 ㎖)로 세척하였다. 여과물을 EtOAc (50 ㎖)로 희석하고 빙냉 0.05 M pH 7.4 인산염 완충 수용액(2 × 50 ㎖), 물 (1 × 50 ㎖) 및 염수(50 ㎖)로 세척하여 건조(MgSO₄) 및 농축시켰다. 잔사를 헥산, CH₂Cl₂/헥산(1/1), CH₂Cl₂, 10% EtOAc/CH₂CL₂및 25% EtOAc/CH₂Cl₂로 연속 추출함으로써 실리카겔(25 g) 컬럼에 통과시켜 표제물질(300 ㎎, 38%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1- in CH ₃ CN (25 mL) Treated with cold solution (ice bath) of azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (760 mg, 1.57 mmol) with chloromethylphenyl sulfide (0.255 mL, 1.9 mmol) This mixture was stirred for 30 minutes. Chloromethylphenyl sulfide (0.5 mL, 3.8 mmol) was added twice, followed by stirring for 30 minutes and 45 minutes, respectively. The cold mixture was passed through a pad of silica gel (5 g) and the pad was washed with cold EtOAc (20 mL). The filtrate was diluted with EtOAc (50 mL) and washed with ice cold 0.05 M pH 7.4 phosphate buffered aqueous solution (2 × 50 mL), water (1 × 50 mL) and brine (50 mL) to dry (MgSO ₄ ) and concentrated. . The residue was passed through a silica gel (25 g) column by successive extraction with hexane, CH ₂ Cl ₂ / hexanes (1/1), CH ₂ Cl ₂ , 10% EtOAc / CH ₂ CL ₂ and 25% EtOAc / CH ₂ Cl ₂ . The title compound (300 mg, 38%) was obtained.

[B 단계][Step B]

포타슘 또는 소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(페닐티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Id)Potassium or sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(phenylthio) methyl] thio] -7-oxo-1-azabicyclo -[3.2.0] hep-2-ten-2-carboxylate (Id)

0.05 M pH 7.0 인산염 완충용액(10 ㎖), 에테르(10 ㎖) 및 THF(10 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(페닐티오)메틸]티오]-7-옥소-1-아자비시클로-[3.2.2] 헵-2-텐-2-카르복실레이트 (230 ㎎, 0.46 m㏖)의 용액을 40 psi H₂에서 1시간동안 10% Pd/C 촉매(230 ㎎)을 이용하여 가수소분해시켰다. 촉매를 여과하였다. 유기상을 분리하고 10% Pd/C 촉매(230 ㎎)로 pH 7.0 완충액(10 ㎖) 이용하여 40 psi에서 1시간동안 더 가수소분해시켰다. 동일 공정을 반복하고 가수소분해 반응말기에 3 수성상을 결합하고, 에테르(2 × 10 ㎖) 세척한 후 C₁₈BondaPak 역상컬럼(23 g 충전재; 컬럼은 먼저 H₂O, 이어서 5%, 10% 및 20% CH₃CN/H₂O로 용출한다)에 통과시켜 표제물질(120 ㎎, 66%)을 얻었다: T¹/₂=100 시간 (pH 7.4), T¹/₂=18.8 분 (pH 2); 순도: 99.9% (HPLC에 의해 측정).P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl]-in 0.05 M pH 7.0 phosphate buffer (10 mL), ether (10 mL) and THF (10 mL) 4-methyl-3-[[(phenylthio) methyl] thio] -7-oxo-1-azabicyclo- [3.2.2] hep-2-ten-2-carboxylate (230 mg, 0.46 mmol) The solution of was hydrolyzed at 40 psi H ₂ using 10% Pd / C catalyst (230 mg) for 1 hour. The catalyst was filtered off. The organic phase was separated and further hydrolyzed at 40 psi for 1 hour using a pH 7.0 buffer (10 mL) with 10% Pd / C catalyst (230 mg). Repeat the same process and combine 3 aqueous phases at the end of the hydrolysis reaction, wash with ether (2 × 10 mL) and then C ₁₈ BondaPak reversed phase column (23 g filler; column first with H ₂ O, then 5%, 10 %, and 20% CH ₃ and eluting with CN / H ₂ O) was passed through to give the title material (120 ㎎, 66%): T 1/2 = 100 sigan ^{(pH 7.4), T 1/} 2 = 18.8 min ( pH 2); Purity: 99.9% (measured by HPLC).

[실시예 8]Example 8

소듐 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ie)Sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ie)

[A 단계][Step A]

p-니트로벤질 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-2-yl) thio] methyl] thio] -7-oxo-1- Azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (2 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (235 ㎎, 0.500 m㏖)의 냉용액(아이스 배쓰)를 DMF (0.5 ㎖)중 2-[(클로로메틸]티오]피리딘(96 ㎎, 0.60 m㏖) 용액으로 처리하고, 이어서 LiI (134 ㎎, 1.00 m㏖) 및 DIPEA (0.10 ㎖, 0.60 m㏖)를 첨가하였다. 혼합물을 실온 약 22℃에서 25분간 교반하고 빙냉수(2.5 ㎖) 및 EtOAc(2.5 ㎖)로 희석후 여과하였다. 여과물을 H₂O (5 ㎖) 및 EtOAc(5 ㎖)로 희석하고 수층을 EtOAc (4 × 10 ㎖)로 추출하였다. 유기층(추출물)을 결합하고, 냉수(5 × 10 ㎖), 및 염소(1 × 10 ㎖)로 세척후 건조하였다(MgSO₄). 잔사를 CCl₄로 분쇄하여 암베이지색 고체를 얻고 이를 예비 TLC 플레이트 상에서 정제하여(EtOAc/CH₂Cl₂:1/1) 표제물질(160 ㎎, 66%)을 얻었다.P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] in DMF (2 mL) A cold solution (ice bath) of hept-2-ten-2-carboxylate (235 mg, 0.500 mmol) was added 2-[(chloromethyl] thio] pyridine (96 mg, 0.60 mmol in DMF (0.5 mL). ) Solution, followed by LiI (134 mg, 1.00 mmol) and DIPEA (0.10 mL, 0.60 mmol) The mixture was stirred at room temperature about 22 ° C. for 25 min and ice cold water (2.5 mL) and EtOAc ( 2.5 mL) and filtered The filtrate was diluted with H ₂ O (5 mL) and EtOAc (5 mL) and the aqueous layer was extracted with EtOAc (4 × 10 mL) The organic layers (extract) were combined and cold water (5 × 10 mL), and washed with chlorine (1 × 10 mL) and dried (MgSO ₄ ) The residue was triturated with CCl ₄ to give an dark beige solid which was purified on a preparative TLC plate (EtOAc / CH _2). Cl ₂ : 1/1) to give the title material (160 mg, 66%).

[B 단계][Step B]

0.05 M, pH 7.0 NaH₂PO₄/NaOH 완충액(80 ㎖, 4 m㏖), 에테르(50 ㎖) 및 THF (50 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (880 ㎎, 1.82 m㏖) 용액을 촉매로서 10% Pd/C(880 ㎎)을 이용하여 15℃에서 1시간동안 45 psi H₂에서 가수소분해시켰다. 촉매를 여과제거하고 H₂O (25 ㎖)로 세척하였다. 유기상을 H₂O (2 × 20 ㎖)로 추출하였다. 수성상을 결합하고, 에테르(2 × 25 ㎖)로 세척한다음 C₁₈μBondaPak 역상컬럼(충전재 75 g; 컬럼은 먼저 H₂O로, 이어서 2%, 5% 및 8% CH₃CN/H₂O로 용출하였다)에 통과시켜 표제화합물(320 ㎎, 47%)을 얻었다: 순도: 99.9% (HPLC로 측정); T¹/₂19시간 (pH 7.4).P-nitrobenzyl (5R, 6S) -6- [1 '(R) in 0.05 M, pH 7.0 NaH ₂ PO ₄ / NaOH buffer (80 mL, 4 mmol), ether (50 mL) and THF (50 mL) ) -Hydroxyethyl] -3-[[[((pyridin-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-car A carboxylate (880 mg, 1.82 mmol) solution was hydrolyzed at 45 psi H ₂ for 1 hour at 15 ° C. using 10% Pd / C (880 mg) as catalyst. The catalyst was filtered off and washed with H ₂ O (25 mL). The organic phase was extracted with H ₂ O (2 × 20 mL). Combine aqueous phase, wash with ether (2 × 25 mL), then C ₁₈ μBondaPak reversed phase column (75 g of filler; column first with H ₂ O, then 2%, 5% and 8% CH ₃ CN / H ₂ Eluted with O) to give the title compound (320 mg, 47%): Purity: 99.9% (measured by HPLC); T ^_1/2 19 sigan (pH 7.4).

[실시예 9]Example 9

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (If)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-2-yl) thio] methyl] thio] -7-oxo -1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (If)

[A 단계][Step A]

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-2-yl) thio] methyl] thio]- 7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (7 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (788 ㎎, 1.62 m㏖) 용액을 5℃에서(아이스 배쓰) DMF (1 ㎖)중 2-[(클로로메틸)티오]피리딘(311 ㎎, 1.95 m㏖)로 처리하고 이어서 LiI (434 ㎎, 3.24 m㏖) 및 DIPEA (0.32 ㎖, 1.95 m㏖)을 첨가하였다. 약 22℃에서 혼합물을 24시간 교반하고, 에틸아세테이트(25 ㎖)와 빙냉 H₂O (25 ㎖)로 희석하였다. 염을 여과 젝하고 EtOAc (10 ㎖)로 세척하였다. 2 용액상을 분리하고 수층을 에틸아세테이트( 3 × 10 ㎖)로 추출하였다. 유기층을 결합시켜 빙냉수(5 × 20 ㎖) 및 염수(1 × 20 ㎖)로 세척후 건조시켰다(MgSO₄). 잔사를 실리카겔(15 g) 컬럼(먼저 CH₂Cl₂로, 이어서 5%, 10%, 15% 및 25% EtOAc/CH₂Cl₂로 용출)에 통과시켜 표제물질을 얻고 이를 예비 TLC 플레이트상에서 재정제하여 (CH₂Cl₂/EtOAc:1/2로 용출) 순수한 생성물 440 ㎎ (54%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo in DMF (7 mL) -[3.2.0] Hep-2-ten-2-carboxylate (788 mg, 1.62 mmol) solution was added 2-[(chloromethyl) thio] pyridine in DMF (1 mL) at 5 ° C. (ice bath). (311 mg, 1.95 mmol) followed by LiI (434 mg, 3.24 mmol) and DIPEA (0.32 mL, 1.95 mmol). The mixture was stirred at about 22 ° C. for 24 hours and diluted with ethyl acetate (25 mL) and ice cold H ₂ O (25 mL). The salt was filtered off and washed with EtOAc (10 mL). 2 solution phases were separated and the aqueous layer was extracted with ethyl acetate (3 × 10 mL). The organic layers were combined, washed with ice cold water (5 × 20 mL) and brine (1 × 20 mL) and dried (MgSO ₄ ). The residue was passed through a silica gel (15 g) column (eluted with CH ₂ Cl ₂ first, followed by 5%, 10%, 15% and 25% EtOAc / CH ₂ Cl ₂ ) to afford the title material which was recrystallized on a preparative TLC plate. Except (eluted with CH ₂ Cl ₂ / EtOAc: 1/2) to give 440 mg (54%) of the pure product.

[B 단계][Step B]

pH 7.0 인산염 완충액(0.05 M, 35 ㎖), 에테르(35 ㎖) 및 THF (35 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (435 ㎎, 0.868 m㏖) 용액을 촉매로서 10% Pd/C (435 ㎎)을 이용하여 15-22 ℃에서 45 psi H₂로 1시간동안 가수소분해시켰다. 혼합물을 에테르(20 ㎖)로 희석하고 촉매를 여과 제거하여 H₂O(10 ㎖)로 세척하였다. 2 용액상을 분리하고 에테르 층을 H₂O (2 × 5 ㎖)로 추출하였다. 수층을 결합시키고, 에테르(3 × 20 ㎖) 세척하여 C₁₈μBondaPak 역상 컬럼에 통과시켜(C₁₈μBondaPak 역상재료 30 g; 컬럼은 먼저 H₂O, 이어서 2%, 5%, 8% 및 10% CH₃CN/H₂O로 연속 용출시킨다) 목탄으로 오염된 표제물질을 얻었다. 생성물을 동결건조시키고 얻어진 분말을 H₂O에 테이크업하여 역상 C₁₈μBondaPak의 스몰패드에 통과시켜(역상 C₁₈μBondaPak 재료 5 g; 컬럼을 먼저 H₂O로, 이어서 2%, 5%, 및 10% CH₃CN/H₂O로 연속 용출함) 순수한 표제물질을 얻는다(85 ㎎, 25%); T¹/₂: 79시간(pH 7.4); T¹/₂36분(pH 2); 순도; 99.2% (298 ㎚에서 HPLC로 측정).p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl]-in pH 7.0 phosphate buffer (0.05 M, 35 mL), ether (35 mL) and THF (35 mL) 4-methyl-3-[[[((pyridin-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate ( 435 mg, 0.868 mmol) solution was hydrolyzed at 45-psi H ₂ at 15-22 ° C. for 1 hour using 10% Pd / C (435 mg) as catalyst. The mixture was diluted with ether (20 mL) and the catalyst was filtered off and washed with H ₂ O (10 mL). The two solution phases were separated and the ether layer was extracted with H ₂ O (2 × 5 mL). The aqueous layers were combined and washed with ether (3 × 20 mL) and passed through a C ₁₈ μBondaPak reversed phase column (30 g of C ₁₈ μBondaPak reversed phase material; the column was first H ₂ O followed by 2%, 5%, 8% and 10% Elution with CH ₃ CN / H ₂ O)). The title material contaminated with charcoal was obtained. The lyophilized product was take-up of the obtained powder in H ₂ O and passed through a small pad of reversed phase C ₁₈ μBondaPak (5 g reversed-phase C ₁₈ μBondaPak material; the column first with H ₂ O, followed by 2%, 5%, and Eluting continuously with 10% CH ₃ CN / H ₂ O) to afford the pure title (85 mg, 25%); T ^_1/2: 79 sigan (pH 7.4); T ^_1/2 36 bun (pH 2); water; 99.2% (measured by HPLC at 298 nm).

[실시예 10]Example 10

포타슘 또는 소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ig)Potassium or sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-4-yl) thio] methyl] thio] -7 Oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (Ig)

[A 단계][Step A]

4-[[클로로메틸)티오]피리딘4-[[chloromethyl) thio] pyridine

벤젠(40 ㎖)중 4-메틸티오피리딘(2.28 g, 18.2 m㏖), N-클로로숙신이미드(3.0 g, 23 m㏖) 및 피리딘(2.9 ㎖, 3.0 m㏖) 용액을 50-60℃에서 3시간 가열하였다. 혼합물을 실온으로 냉각시켰다. 용매를 따라내고 잔사를 벤젠(2 × 10 ㎖)으로 헹구었다. 유기층을 결합시켜 증발시켰다. 잔사를 실리카겔(40 g) 패드로 통과시켜 출발물질로 오염된 표제물질(1.35 g)을 얻었다. 실리카겔 예비 플레이트에서 이 불순한 생성물을 재정제하여(CH₂Cl₂/CH₃CN:1/1) 순수한 표제물질(926 ㎎, 32%)을 황색오일로서 얻었다.A solution of 4-methylthiopyridine (2.28 g, 18.2 mmol), N-chlorosuccinimide (3.0 g, 23 mmol) and pyridine (2.9 mL, 3.0 mmol) in benzene (40 mL) was heated at 50-60 ° C. Heated for 3 h. The mixture was cooled to room temperature. The solvent was decanted and the residue was rinsed with benzene (2 x 10 mL). The organic layers were combined and evaporated. The residue was passed through a pad of silica gel (40 g) to give the titled material (1.35 g) contaminated with starting material. This impure product was repurchased in a silica gel prep plate (CH ₂ Cl ₂ / CH ₃ CN: 1/1) to give pure title (926 mg, 32%) as a yellow oil.

[B 단계][Step B]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-4-yl) thio] methyl] thio]- 7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (8 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (794 ㎎, 1.00 m㏖), 4-[(클로로메틸)티오]피리딘(314 ㎎, 1.97 m㏖), LiI (440 ㎎, 3.28 m㏖) 및 DIPEA (0.345 ㎖, 2.0 m㏖) 용액을 약 22℃에서 20시간 교반하였다. 혼합물을 에틸아세테이트(10 ㎖) 및 물(5 ㎖)로 희석하고 여과하였다. 수상을 에틸아세테이트(2 × 5 ㎖)로 추출하였다. 유기추출물을 결합하고, 냉수(4 × 10 ㎖) 및 염수(1 × 10 ㎖)로 세척하고 건조(MgSO₄)후 농축시켰다. 예비 TLC 플레이트상에서 잔사를 정제하여 (10% CH₃CN/EtOAc로 용출) 표제물질(430 ㎎, 52%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo in DMF (8 mL) -[3.2.0] hep-2-ten-2-carboxylate (794 mg, 1.00 mmol), 4-[(chloromethyl) thio] pyridine (314 mg, 1.97 mmol), LiI (440 mg, 3.28 mmol) and DIPEA (0.345 mL, 2.0 mmol) solution were stirred at about 22 ° C. for 20 hours. The mixture was diluted with ethyl acetate (10 mL) and water (5 mL) and filtered. The aqueous phase was extracted with ethyl acetate (2 × 5 mL). The organic extracts were combined, washed with cold water (4 × 10 mL) and brine (1 × 10 mL), dried (MgSO ₄ ) and concentrated. The residue was purified on a preparative TLC plate (eluted with 10% CH ₃ CN / EtOAc) to give the title material (430 mg, 52%).

[C 단계][Step C]

pH 7.0 NaH₂PO₄/NaOH 또는 KH₂PO₄/KOH 완충액(0.05 M, 30 ㎖), 에테르(30 ㎖) 및 THF(30 ㎖)중 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-4-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (663 ㎎, 1.32 m㏖) 용액을 40 psi H₂에서 10% Pd/C 촉매(663 ㎎)로 1시간 가수소분해시켰다. 촉매를 pH 7.0 완충액(0.05 M, 5㎖) 및 물(5㎖)로 세척하였다. 수층을 에테르(3 × 20 ㎖)로 세척하고 C₁₈μ-BondaPak 역상컬럼에 통과시켜(C₁₈μ-BondaPak 역상물질 55 g) 표제화합물 282 ㎎ (54%)을 황색고체로서 얻었다: T¹/₂=34분 (pH 2); 순도 99.9%, HPLC로 측정(체류시간, 9.63분, 10% CH₃CN/pH 7 완충액).p-nitrobenzyl (4R, 5S, 6S) -6- in pH 7.0 NaH ₂ PO ₄ / NaOH or KH ₂ PO ₄ / KOH buffer (0.05 M, 30 mL), ether (30 mL) and THF (30 mL) [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (663 mg, 1.32 mmol) solution was hydrolyzed for 1 hour with 10% Pd / C catalyst (663 mg) in 40 psi H ₂ . The catalyst was washed with pH 7.0 buffer (0.05 M, 5 mL) and water (5 mL). Wash the aqueous layer with ether (3 × 20 ㎖) and C ₁₈ μ-BondaPak passed through a reverse phase column (C ₁₈ μ-BondaPak reversed-phase material 55 g) to give the title compound 282 ㎎ (54%) as a yellow solid: T ¹ / ₂ = 34 min (pH 2); Purity 99.9%, determined by HPLC (ret. Time, 9.63 min, 10% CH ₃ CN / pH 7 buffer).

[실시예 11]Example 11

소듐 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-3-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ih)Sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] Hep-2-ten-2-carboxylate (Ih)

[A 단계][Step A]

p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-3-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo-1- Azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (30 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (3.0 g, 6.37 m㏖)의 냉용액(아이스 배쓰) DIPEA (1.27 ㎖, 7.65 m㏖), LiI(1.7 g, 12.74 m㏖) 및 DMF (10 ㎖)중 3-[(클로로메틸)티오]-피리딘(1.22 g, 7.65 m㏖)을 첨가하였다. 이 혼합물을 약 22℃에서 24시간 교반한다음, 빙냉수(500 ㎖) 및 EtOAc (5 × 200 ㎖)로 헹구고 상을 리하였다. 수층을 EtOAc (2 × 50 ㎖)로 추출하였다. 유기층을 결합하여 냉수(4 × 250 ㎖) 및 냉염수(250 ㎖)로 세척하고 활성목탄(중성)으로 처리후 건조시켰다(MgSO₄). 용매증발후 얻어진 고체 잔사를 CH₂Cl₂/에테르(9.5/0.5, 40 ㎖)로 분쇄하고, 여과수집하여 에테르(10 ㎖)로 헹구어 표제물질 1.17 (55%)을 얻었다.P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] in DMF (30 mL) Hep-2-ten-2-carboxylate (3.0 g, 6.37 mmol) cold solution (ice bath) DIPEA (1.27 mL, 7.65 mmol), LiI (1.7 g, 12.74 mmol) and DMF (10 mL) 3-[(chloromethyl) thio] -pyridine (1.22 g, 7.65 mmol) was added. The mixture was stirred at about 22 ° C. for 24 hours, then rinsed with ice cold water (500 mL) and EtOAc (5 × 200 mL) and the phases were separated. The aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with cold water (4 × 250 mL) and cold brine (250 mL), treated with activated charcoal (neutral) and dried (MgSO ₄ ). The solid residue obtained after evaporation of the solvent was triturated with CH ₂ Cl ₂ / ether (9.5 / 0.5, 40 mL), collected by filtration and rinsed with ether (10 mL) to obtain the titled substance 1.17 (55%).

[B 단계][Step B]

0.05 M pH 7.0 NaH₂PO₄/NaOH 완충수용액 (30 ㎖, 1.5 m㏖), 에테르(30 ㎖) 및 THF (30 ㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-2-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (400 ㎎, 0.82 mmole) 용액을 촉매로서 10% Pd/C (400 ㎎)을 이용, 10℃ 내지 약 22℃에서 45 psi H₂에서 1시간동안 Parr 수소첨가기에서 와동시켰다. 혼합물을 에테르(30 ㎖)로 희석하고 촉매를 여과 제거하였다. 이를 pH 7.0 완충액(2 × 5 ㎖)로 헹구었다. 유기층을 분리하고 완충액(2 × 25 ㎖)로 추출하였다. 수상을 결합하고 에테르(2 × 25 ㎖) 세척한 후 C₁₈μBondaPak 역상컬럼에 통과시켜(C₁₈μBondaPak 역상재료 30 g; 먼저 H₂O로, 이어서 2%, 5%, 8% 및 12% CH₃CN/H₂O로 연속 용출) 표제물질(120 ㎎, 39%)을 동결건조 분말로서 얻었다; 순도 99.3% (HPLC로 측정); T¹/₂22시간 (pH 7.4), T¹/₂2분 (pH 2).P-nitrobenzyl (5R, 6S) -6- [1 '(R in 0.05 M pH 7.0 NaH ₂ PO ₄ / NaOH buffer (30 mL, 1.5 mmol), ether (30 mL) and THF (30 mL) ) -Hydroxyethyl] -3-[[[((pyridin-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-car Voxylate (400 mg, 0.82 mmole) solution was vortexed in a Parr hydrogenator for 1 hour at 45 psi H ₂ at 10 ° C. to about 22 ° C. using 10% Pd / C (400 mg) as catalyst. The mixture was diluted with ether (30 mL) and the catalyst was filtered off. It was rinsed with pH 7.0 buffer (2 × 5 mL). The organic layer was separated and extracted with buffer (2 × 25 mL). Combine the aqueous phase and wash with ether (2 × 25 mL) and pass through a C ₁₈ μBondaPak reversed phase column (30 g C ₁₈ μBondaPak reversed phase material; first with H ₂ O, then 2%, 5%, 8% and 12% CH). Continuous elution with ₃ CN / H ₂ O) The title material (120 mg, 39%) was obtained as a lyophilized powder; Purity 99.3% (measured by HPLC); T ^_1/2 22 sigan ^{(pH 7.4), T 1/} 2 2 bun (pH 2).

[실시예 12]Example 12

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-3-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (Ii)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo -1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate (Ii)

[A 단계][Step A]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-3-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-3-yl) thio] methyl] thio]- 7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2-carboxylate

DMF (15 ㎖)중 p-니트로벤질(4R,5S,6S)-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (1.7 g, 3.5 m㏖) 냉용액(아이스 배쓰)에 3-[(클로로메틸)티오]피리딘(670 ㎎, 4.2 m㏖) 이어서 LiI (940 ㎎, 7.0 m㏖) 및 DIPEA (0.69 ㎖, 4.2 m㏖)을 첨가하였다. 약 22℃에서 혼합물을 22시간 교반한다음 빙냉수 및 EtOAc로 희석하고 Celite 패드를 통해 여과시켰다. 패드를 EtOAc (4 × 20 ㎖)로 헹구고 수층을 EtOAc (2 × 40 ㎖)로 추출하였다. 유기 추출물을 결합하고 냉수(4 × 25 ㎖) 및 염수로 세척후 건조(MgSO₄)하고 농축시켰다. 잔사를 실리카겔 플래쉬 컬럼(실리카 30 g; 컬럼은 CH₂Cl₂, 10%, 25% 및 50% CH₂Cl₂/EtOAc, EtOAc, 10% 및 20% CH₃CN/EtOAc로 연속 용출시킴)에 통과시켜 표제물질(804 ㎎, 46%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo- [3.2.0] hep-2-ten-2- in DMF (15 mL) To carboxylate (1.7 g, 3.5 mmol) cold solution (ice bath) was added 3-[(chloromethyl) thio] pyridine (670 mg, 4.2 mmol) followed by LiI (940 mg, 7.0 mmol) and DIPEA (0.69 Ml, 4.2 mmol) was added. The mixture was stirred for 22 h at about 22 ° C., then diluted with ice cold water and EtOAc and filtered through a Celite pad. The pad was rinsed with EtOAc (4 × 20 mL) and the aqueous layer was extracted with EtOAc (2 × 40 mL). The organic extracts were combined, washed with cold water (4 × 25 mL) and brine, dried (MgSO ₄ ) and concentrated. The residue was subjected to silica gel flash column (30 g of silica; column eluted continuously with CH ₂ Cl ₂ , 10%, 25% and 50% CH ₂ Cl ₂ / EtOAc, EtOAc, 10% and 20% CH ₃ CN / EtOAc). Passing gave the title material (804 mg, 46%).

[B 단계][Step B]

0.05 M pH 7.0 수성 NaH₂PO₄/NaOH 완충액(50 ㎖, 2.5 m㏖), 에테르(45 ㎖) 및 THF(45 ㎖)중 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리디-3-닐)티오]메틸]티오]-7-옥소-1-아자비시클로-[3.2.0] 헵-2-텐-2-카르복실레이트 (725 ㎎, 1.44 m㏖) 용액을 촉매로서 10% Pd/C (725 ㎎)을 사용하여 15 내지 22℃, 45 psi H₂에서 1 시간동안 Parr 수소첨가기내에서 와동시켰다. 혼합물을 에테르(50 ㎖)로 희석하였다. 촉매를 여과 제거하고 pH 7.0 완충액 (2 × 10 ㎖)로 헹구었다. 2 용액상을 분리하고 유기층을 pH 7.0 완충수용액(1 × 10 ㎖)로 추출하였다. 수층을 결합하고, 에테르(2 × 25 ㎖)로 세척하여 역상 C₁₈μBondaPak 컬럼 (역상 C₁₈μBondaPak 재료 75 g; 컬럼은 먼저 H₂O, 이어서 2%, 5%, 10% 및 15% CH₃CN/H₂O로 연속 용출함)에 통과시켜 표제물질(220 ㎎, 39%)을 얻었다; 순도 99.6% (HPLC로 측정); T¹/₂74시간(pH 7.4); T¹/₂35분 (pH 2.0).P-nitrobenzyl (4R, 5S, 6S) -6- [1 'in 0.05 M pH 7.0 aqueous NaH ₂ PO ₄ / NaOH buffer (50 mL, 2.5 mmol), ether (45 mL) and THF (45 mL) (R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo-1-azabicyclo- [3.2.0] hep-2 -Ten-2-carboxylate (725 mg, 1.44 mmol) solution in Parr hydrogenator for 1 hour at 15-22 ° C., 45 psi H ₂ using 10% Pd / C (725 mg) as catalyst Vortex The mixture was diluted with ether (50 mL). The catalyst was filtered off and rinsed with pH 7.0 buffer (2 × 10 mL). 2 solution phases were separated and the organic layer was extracted with pH 7.0 buffered aqueous solution (1 × 10 mL). Combine the aqueous layers and wash with ether (2 × 25 mL) to reverse phase C ₁₈ μBondaPak column (75 g of reverse phase C ₁₈ μBondaPak material; column first with H ₂ O, then 2%, 5%, 10% and 15% CH ₃ Elution with CN / H ₂ O) yielded the title material (220 mg, 39%); Purity 99.6% (measured by HPLC); T ^_1/2 74 sigan (pH 7.4); T ^_1/2 35 bun (pH 2.0).

[실시예 13]Example 13

소듐 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(Ij)Sodium (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(p-chlorophenyl) thio] -methyl] thio] -7-oxo-1-azabicyclo [3.2 .0] hep-2-ten-2-carboxylate (Ij)

[A단계][Step A]

p-니트로벤질 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트p-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(p-chlorophenyl) thio] -methyl] thio] -7-oxo-1-azabi Cyclo [3.2.0] hep-2-ten-2-carboxylate

DMF(6㎖)중 p-니트로벤질(5R,6S)-6-[1'(R)-히드록시에틸]-3-실버 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(471㎎, 1m㏖)의 냉현탁액(아이스 배쓰)에 클로로메틸 4-클로로페닐 술파이드(0.18㎖, 1.3m㏖) LiI(267㎎, 2m㏖)및 DIPEA(0.20㎖, 1.2m㏖)을 첨가하였다. 얻어진 혼합물을 18시간 교반한 다음, 냉수(12㎖) 및 EtOAc(6 × 15㎖)로 세척하고 그 용액상을 분리하였다. 수성상을 EtOAc(3 × 15㎖)로 추출하였다. 유기 추출물을 결합하고, 냉 H₂O(2 × 25㎖) 및 염수(25㎖)로 세척후 건조하였다(MgSO₄). 용매증발 후 얻은 고체 잔사를 에테르/CH₂Cl₂혼합물(9/11, 20㎖)로 분쇄하여 표제 물질(355㎎, 68%)을 담갈색 고체로서 얻었다.P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-silver mercapto-7-oxo-1-azabicyclo [3.2.0] hep in DMF (6 mL) In a cold suspension (ice bath) of -2-ten-2-carboxylate (471 mg, 1 mmol), chloromethyl 4-chlorophenyl sulfide (0.18 mL, 1.3 mmol) LiI (267 mg, 2 mmol), and DIPEA (0.20 mL, 1.2 mmol) was added. The resulting mixture was stirred for 18 hours, then washed with cold water (12 mL) and EtOAc (6 × 15 mL) and the solution phase was separated. The aqueous phase was extracted with EtOAc (3 x 15 mL). The organic extracts were combined, washed with cold H ₂ O (2 × 25 mL) and brine (25 mL) and dried (MgSO ₄ ). The solid residue obtained after solvent evaporation was triturated with an ether / CH ₂ Cl ₂ mixture (9/11, 20 mL) to give the title material (355 mg, 68%) as a light brown solid.

[B단계][Step B]

0.05M 인산염 완충액(pH 7.0, 64㎖, 3.2m㏖), 에테르(40㎖) 및 THF(40㎖)중 p-니트로벤질 (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(900㎎, 1.73m㏖)용액을 촉매로서 10% Pd/C(2 × 900㎎)을 이용하여 45-50 psi H₂에서 3시간 동안 Parr 수소첨가에서 와동시켰다. 촉매를 여과시켜 제거하고 pH 7.0용액 (2 × 10㎖)으로 세척하였다. 유기상을 pH 7.0 용액 (2 × 10㎖)으로 추출하였다. 수성 추출물을 결합하고, 에테르 (2 × 50 ㎖)로 세척후 C₁₈μBondaPak 역상 컬럼에 통과시켜 (C₁₈μBondaPak 역상물질 90g; 컬럼은 먼저 H₂O, 이어서 5%, 10% 및 20% CH₃CN/H₂O로 연속 용출시킨다.) 표제물질을 얻고 이를 동일 유형의 컬럼(역상 물질 20g)상에서 재정제하여 최종적으로 순수한 물질을 우수한 수율로 얻었다(212㎎, 30%); 순도; 99.4%(HPLC로 측정); T¹/₂26시간(pH 7.4).P-nitrobenzyl (5R, 6S) -6- [1 '(R) -hydroxyethyl in 0.05M phosphate buffer (pH 7.0, 64 mL, 3.2 mmol), ether (40 mL) and THF (40 mL) ] -3-[[[(p-chlorophenyl) thio] -methyl] thio] -7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (900 mg, 1.73) mmol) solution was vortexed with Parr hydrogenation for 3 hours at 45-50 psi H ₂ using 10% Pd / C (2 × 900 mg) as catalyst. The catalyst was filtered off and washed with pH 7.0 solution (2 × 10 mL). The organic phase was extracted with pH 7.0 solution (2 × 10 mL). The aqueous extracts were combined, washed with ether (2 × 50 mL) and passed through a C ₁₈ μBondaPak reversed phase column (90 g of C ₁₈ μBondaPak reversed phase; column firstly H ₂ O followed by 5%, 10% and 20% CH _3). Continuous eluting with CN / H ₂ O.) The title material was obtained and recrystallized on the same type of column (20 g of reversed phase material) to finally give a pure material in good yield (212 mg, 30%); water; 99.4% (as measured by HPLC); T ^_1/2 26 sigan (pH 7.4).

[실시예 14]Example 14

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(Ik)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-chlorophenyl) thio] -methyl] thio] -7-oxo- 1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (Ik)

[A단계][Step A]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-chlorophenyl) thio] -methyl] thio] -7 Oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate

DMF(15㎖)중 p-니크로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-실버 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트 (1.5g, 3.1 m㏖)의 냉현탁액(아이스 배쓰)를 클로로메틸 4-클로로페닐 술파이드 (0.56㎖, 3.9m㏖)로 처리한 다음 LiI (422㎎, 6.2m㏖) 및 DIPEA(0.62㎖, 3.72m㏖)을 첨가하였다. 얻어진 혼합물을 약22℃에서 23시간 교반한 다음 냉수(50㎖) 및 EtOAc(50㎖)로 희석하고 여과하여 고체를 제거하였다. 이 고체를 EtOAc로 헹궜다.(2 × 20㎖). 수상을 분리하고 EtOAc (2× 20㎖)로 추출하였다. 유기상을 결합시키고, 냉수 (2 × 50㎖) 및 염수 (1 × 50㎖)로 세척하고, 건조(MgSO₄) 후 농축하였다. 전사를 실리카 겔의 실리카 겔 플래쉬 컬럼(30g)을 통해 통과시켜 (실리카 겔 30g; 컬럼은 먼저 CH₂Cl₂, 이어서 10%, 20%, 및 30% EtOAc/CH₂Cl₂로 용출시켰다). 표제물질 (819㎎, 49%)을 무정형 담황색 고체로서 얻었다.P-nicrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-silver mercapto-7-oxo-1-azabicyclo in DMF (15 mL) [3.2.0] cold suspension (ice bath) of hep-2-ten-2-carboxylate (1.5 g, 3.1 mmol) was treated with chloromethyl 4-chlorophenyl sulfide (0.56 mL, 3.9 mmol) Then LiI (422 mg, 6.2 mmol) and DIPEA (0.62 mL, 3.72 mmol) were added. The resulting mixture was stirred at about 22 ° C. for 23 hours and then diluted with cold water (50 mL) and EtOAc (50 mL) and filtered to remove solids. This solid was rinsed with EtOAc (2 × 20 mL). The aqueous phase was separated and extracted with EtOAc (2 × 20 mL). The organic phases were combined, washed with cold water (2 × 50 mL) and brine (1 × 50 mL), dried (MgSO ₄ ) and concentrated. Transfer was passed through a silica gel flash column (30 g) of silica gel (silica gel 30 g; column eluted first with CH ₂ Cl ₂ , then 10%, 20%, and 30% EtOAc / CH ₂ Cl ₂ ). The title material (819 mg, 49%) was obtained as an amorphous pale yellow solid.

[B단계][Step B]

pH 7.0 NaH₂PO₄/NaOH 완충액 (0.05M, 56㎖, 2.8 m㏖), 에테르(40㎖) 및 THF(40㎖)중 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-클로로페닐)티오]-메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(800㎎, 1.49m㏖)용액을 촉매로서 10% Pd/C (800㎎)을 이용하여 45psi H₂에서 3시간 동안 Parr 수소첨가기에서 와동시켰다. 촉매를 Celite케익을 통해여과시켜 제거하고 케익은 pH 7.0 완충액 (2 × 20㎖)로 헹구었다. 유기성을 분리하고 0.05M pH 7.0 인산염 완충액 (2 × 20㎖)로 추출하였다. 수층을 결합시키고, 에테르(2 × 50㎖)로 세척후 C₁₈μBondaPak 역상 컬럼(C₁₈μBondaPak 역상물질 40g; 컬럼은 먼저 H₂O로, 이어서 5%, 10% 및 20% CH₃CN/H₂O로 연속 용출시킴)에 통과시켜 표제물질(315㎎, 50%)를 얻었다: 순도 99.4% (HPLC로 측정); T¹/₂97시간 (pH 7.4).p-nitrobenzyl (4R, 5S, 6S) -6- [1 'in pH 7.0 NaH ₂ PO ₄ / NaOH buffer (0.05M, 56 mL, 2.8 mmol), ether (40 mL) and THF (40 mL) (R) -hydroxyethyl] -4-methyl-3-[[[(p-chlorophenyl) thio] -methyl] thio] -7-oxo-1-azabicyclo [3.2.0] hep-2-ten A 2-carboxylate (800 mg, 1.49 mmol) solution was vortexed in a Parr hydrogenator for 3 hours at 45 psi H ₂ using 10% Pd / C (800 mg) as catalyst. The catalyst was removed by filtration through Celite cake and the cake was rinsed with pH 7.0 buffer (2 × 20 mL). The organics were separated and extracted with 0.05M pH 7.0 phosphate buffer (2 × 20 mL). Combine the aqueous layers, wash with ether (2 × 50 mL) and then C ₁₈ μBondaPak reversed phase column (40 g of C ₁₈ μBondaPak reversed phase; column first with H ₂ O, then 5%, 10% and 20% CH ₃ CN / H Elution with ₂ O) yielded the title material (315 mg, 50%): purity 99.4% (measured by HPLC); T ^_1/2 97 sigan (pH 7.4).

[실시예 15]Example 15

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-메틸술피닐)메틸]-티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(Im)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-methylsulfinyl) methyl] -thio] -7-oxo-1 Azabicyclo [3.2.0] hep-2-ten-2-carboxylate (Im)

H₂O(10㎖)중 소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(Ib')(100㎎, 0.31m㏖) 의 냉용액(아이스 배쓰) H₂O₂30% 수용액 (0.03㎖, 0.31m㏖)을 적가하고 5℃(아이스 배쓰)에서 8.5시간 교반하였다. 다음 용액을 C₁₈μBondaPak 역상 컬럼에 통과시켜(C₁₈μBondaPak 역상물질 10g, 컬럼 H₂O로 용출시킴) 표제화합물을 입체이성체 술폭사이드의 혼합물로서, 일부는 순수 생성물로서[78㎎, 74%, 순도 99.54%(HPLC로 측정)], 일부는 불순 생성물로서 [20㎎, 19%, 순도 97.6%(HPLC로 측정)]얻었다.; 입체 이성체 비: 6/4; T¹/₂46시간 (pH 7.4).Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(methylthio) methyl] thio] -7 in H ₂ O (10 mL) -oxo-1-azabicyclo [3.2.0] heptane-2-X-2-carboxylate (Ib ') a cold solution (ice bath) H _₂ O ₂ 30% aqueous solution of (100㎎, 0.31m㏖) ( 0.03 ml, 0.31 mmol) was added dropwise and stirred at 5 ° C (ice bath) for 8.5 hours. The solution was then passed through a C ₁₈ μBondaPak reversed phase column (eluted with 10 g of C ₁₈ μBondaPak reversed phase, column H ₂ O) to give the title compound as a mixture of stereoisomeric sulfoxides, some as pure product [78 mg, 74%, Purity 99.54% (measured by HPLC)], some obtained as impurity product [20 mg, 19%, purity 97.6% (measured by HPLC)]; Stereoisomeric ratio: 6/4; T ^1/2 ₂ h (pH 7.4).

[실시예 16]Example 16

소듐 (4R,5S,6S)-3-[[[(p-클로로페닐)술피닐]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(In)Sodium (4R, 5S, 6S) -3-[[[(p-chlorophenyl) sulfinyl] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo- 1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (In)

H₂O(15㎖)중 소듐(4R,5S,6S)-3-[[[(p-클로로페닐)티오]-메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵-2-텐-2-카르복실레이트(Ik)(324㎎, 0.77m㏖)의 냉용액(아이스 배쓰)을 H₂O 30% 수용액 (0.087㎖, 0.77m㏖)으로 적가처리하고 5℃에서 2일간 교반하였다. 차가운 이 수용액을 C₁₈μBondaPak 역상 컬럼에 부어(C₁₈μBondaPak 역상물질 30g; 컬럼은 먼저 H₂O, 이어서 2%, 5% 및 10% CH₃CN/H₂O로 연속 용출시킴) 표제물질(158㎎, 47%)을 입체 이성체의 26/74혼합물로서 얻었다; T¹/₂106시간 (pH 7.4, 37℃).Sodium (4R, 5S, 6S) -3-[[[(p-chlorophenyl) thio] -methyl] thio] -6- [1 '(R) -hydroxyethyl]-in H ₂ O (15 mL) A cold solution (ice bath) of 4-methyl-7-oxo-1-azabicyclo [3.2.0] hep-2-ten-2-carboxylate (Ik) (324 mg, 0.77 mmol) was purified by H ₂ O. The solution was added dropwise with 30% aqueous solution (0.087 mL, 0.77 mmol) and stirred at 5 ° C. for 2 days. Pour this cold aqueous solution into a C ₁₈ μBondaPak reversed phase column (30 g of C ₁₈ μBondaPak reversed phase; column eluted first with H ₂ O followed by 2%, 5% and 10% CH ₃ CN / H ₂ O). 158 mg, 47%) was obtained as a 26/74 mixture of stereoisomers; T ^_1/2 106 sigan (pH 7.4, 37 ℃).

[실시예 17]Example 17

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Io)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-pyridin-3- yl) methyl] thio] methyl] thio] 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Io)

[단계 A][Step A]

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-pyridin-3-yl) methyl] thio] methyl] Thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF(6㎖)내 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(1.0g, 2.06m㏖)의 냉각(얼음조) 용액에 새로 제조된 DMF(4㎖)내 3-[[(클로로메틸)티오]메틸]피리딘 (3-피콜릴 메르캅탄 1.24g, 10m㏖로부터) 용액 이어서 LiI(280㎎, 4.12 m㏖) 및 DIPEA(0.41㎖, 2.47m㏖)를 첨가하였다. 얻어진 혼합물을 약22℃에서 18시간 교반한다음, 냉각 H₂O(25㎖) 및 냉각EtOAc(25㎖)로서 희석하였고, 셀라이트 패드로 통과시켰다. 패드를 EtOAc(4 × 10㎖)로 세척한 다음 두개의 용액층을 분리시켰다. 수성상을 EtOAc(2 × 10㎖)로 추출하였고 유기분율을 결합하였다. 그들을 냉각 H₂O (2 × 25㎖) 및 염수(25㎖)로 세척한다음 건조시켰다(MgSO₄). 용매 증발시 얻어진 잔류물을 실리카겔 컬럼(실리카 20g; 컬럼을 처음에 CH₂Cl₂로서 이어서 연속적으로 10%, 20%, 40% 및 60% EtOAc/CH₂Cl₂끝으로 EtOAc로서 추출함)을 통과시켜 황색고체로서 표제의 물질(413㎎, 41%을 제공하였다.)P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1-azabicyclo in DMF (6 mL) 3-[[(chloromethyl) thio in freshly prepared DMF (4 mL) in a cooled (ice bath) solution of [3.2.0] heptan-2-ene-2-carboxylate (1.0 g, 2.06 mmol). ] Methyl] pyridine (from 1.24 g of 3-picolyl mercaptan, 10 mmol) solution followed by LiI (280 mg, 4.12 mmol) and DIPEA (0.41 mL, 2.47 mmol). The resulting mixture was stirred at about 22 ° C. for 18 hours, then diluted with cold H ₂ O (25 mL) and cold EtOAc (25 mL) and passed through a pad of celite. The pad was washed with EtOAc (4 × 10 mL) and then the two solution layers were separated. The aqueous phase was extracted with EtOAc (2 × 10 mL) and the organic fractions combined. They were washed with cold H ₂ O (2 × 25 mL) and brine (25 mL) and dried (MgSO ₄ ). The residue obtained upon evaporation of the solvent was subjected to a silica gel column (20 g of silica; the column was initially extracted as CH ₂ Cl ₂ followed by successively 10%, 20%, 40% and 60% EtOAc / CH ₂ Cl ₂ ends as EtOAc). Passing gave the title material (413 mg, 41%) as a yellow solid.

[단계B][Step B]

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Io)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3-yl) methyl] thio] methyl] thio] -7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Io)

THF(40㎖), 에테르(40㎖) 및 0.05M pH 7.0 NaH₂PO₄/NaOH 완충용액(50㎖, 2.5m㏖)내 p-나트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(718㎎, 1.39m㏖)용액을 촉매로서 10% Pd/C(700㎎)을 이용하여 Parr수소첨가장치에서, 약22℃에서 45psi H₂에서 3시간 교반하였다. 여과에 의하여 촉매를 제거하였고 0.05M pH 7.0 포스페이트 완충용액(2 × 10㎖)으로서 린스하였다. 용액층을 분리하였고 유기층을 0.05M pH 7.0 포스페이트 완충용액으로 추출하였다. 수성층을 결합하였고, 에테르 (2 × 50㎖)로 세척한다음 역상 C₁₈μBondaPak 물질 40g; 컬럼을 처음에 H₂O 이어서 연속적으로 2%, 5%, 10% 및 15% CH₃CN/H₂O로서 용출함)을 통과시켜 친액화후 회색고체(267㎎, 48%)를 제공하였다. 이 고체는 다시 C₁₈μBondaPak 컬럼(C₁₈μBondaPak 물질 7.5g; 컬럼을 연속적으로 H₂O 및 2%와 5% CH₃CN/H₂O로서 용출함)을 통과시켜 백색 친액화 고체로서 표제의 화합물(2.33㎎, 42%)을 제공하였다: T¹/₂77시간 (pH 7.4, 37℃), 순도 99.9% (HPLC에 의하여 검사).P-natrobenzyl (4R, 5S, 6S) -6- [1 in THF (40 mL), ether (40 mL) and 0.05 M pH 7.0 NaH ₂ PO ₄ / NaOH buffer (50 mL, 2.5 mmol). '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3-yl) methyl] thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptane 2-hr-2-en-2-carboxylate (718 mg, 1.39 mmol) solution using a 10% Pd / C (700 mg) as a catalyst, in a Parr hydrogenator, at about 22 ° C. at 45 psi H ₂ for 3 hours Stirred. The catalyst was removed by filtration and rinsed as 0.05 M pH 7.0 phosphate buffer (2 × 10 mL). The solution layer was separated and the organic layer was extracted with 0.05M pH 7.0 phosphate buffer. The aqueous layers were combined, washed with ether (2 × 50 mL) followed by 40 g of reversed phase C ₁₈ μBondaPak material; The column was first passed through H ₂ O followed by successively eluting as 2%, 5%, 10% and 15% CH ₃ CN / H ₂ O) to give a gray solid (267 mg, 48%) after lyophilisation. . This solid was again passed through a C ₁₈ μBondaPak column (7.5 g of C ₁₈ μBondaPak material; eluting the column continuously with H ₂ O and 2% and 5% CH ₃ CN / H ₂ O) as a white lyophilic solid. compound (2.33㎎, 42%) to give a (check by HPLC) T ^_1/2 77 sigan (pH 7.4, 37 ℃), 99.9% purity.

[실시예 18]Example 18

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]술핀일]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Ip)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3- yl) methyl] sulfinyl] methyl] thio]- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Ip)

H₂O(20㎖)내 소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Io)(390㎎, 0.97m㏖) 냉각(얼음조) 용액을 H₂O(5㎖)내 NaIO₄(229㎎, 1.07m㏖) 냉각용액으로 처리하였다. 혼합물을 5℃에서 2시간 교반한 다음, C₁₈μBondaPak 역상 컬럼 (C₁₈μBondaPak 역상물질 50g; 컬럼을 처음에 H₂O 이어서 2%와 5%, CH₃CN/H₂O로서 용출함)을 통과시켜 친액화후 황색고체(210㎎, 52%) 및 극성이 적은 물질(90㎎)을 제공하였다. 이와같은 후자 물질을 냉각 H₂O(5㎖) 내 NaHCO₃(3eq.)으로 처리한 다음 다시 역상 컬럼(역상물질 10g, 컬럼을 H₂O 그후 2% CH₃CN/H₂O로서 용출함)을 통과시켜 표제의 물질(73㎎)을 제공하였다. 이와 같이 얻어진 두개의 분율을 결합하였고 C₁₈μBondaPak 컬럼(역상물질 30g; 컬럼을 연속적으로 H₂O, 2%와 5% CH₃CN/H₂O로 용출함)으로 재크로마토그래피시켜 부분입체이성체의 71/29 혼합물로서 순수한 생성물(169㎎, 32%)을 얻었다: 순도 99.2% (HPLC에 의하여 검사); T¹/₂43시간 (7.4, 37℃).Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3-yl) methyl] in H ₂ O (20 mL) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Io) (390 mg, 0.97 mmol) Cooled (ice bath) solution Treated with NaIO ₄ (229 mg, 1.07 mmol) cooling solution in ₂ O (5 mL). The mixture was stirred at 5 ° C. for 2 hours and then a C ₁₈ μBondaPak reversed phase column (50 g of C ₁₈ μBondaPak reversed phase; the column was initially eluted with H ₂ O followed by 2% and 5%, CH ₃ CN / H ₂ O). Passing gave a yellow solid (210 mg, 52%) and a less polar substance (90 mg) after lyophilisation. This latter material was treated with NaHCO ₃ (3eq.) In cold H ₂ O (5 mL) and again eluted with a reversed phase column (10 g of reversed phase, H ₂ O followed by 2% CH ₃ CN / H ₂ O ) To give the title material (73 mg). The two fractions thus obtained were combined and rechromatated on a C ₁₈ μBondaPak column (reverse phase 30g; eluting the column continuously with H ₂ O, 2% and 5% CH ₃ CN / H ₂ O) to give diastereomers. Pure product (169 mg, 32%) was obtained as a 71/29 mixture of: purity 99.2% (check by HPLC); T ^_1/2 43 sigan (7.4, 37 ℃).

[실시예 19]Example 19

소듐 (4R,5S,6S)-3-[[[(3,4-디클로로페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iq)Sodium (4R, 5S, 6S) -3-[[[((3,4-dichlorophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo -1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iq)

[단계 A][Step A]

p-니트로벤질 (4R,5S,6S)-3-[[[(3,4-디클로로페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[(3,4-dichlorophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF(6㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-은 메르캅토-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (1.00g, 2.06 m㏖)의 냉각(얼음조) 용액에 새로 제조된 1-클로로메틸티오-3,4-디클로벤젠용액 (3,4-디클로로페닐 메르캅탄 1.27㎖, 10.0m㏖로 부터 제조됨)을 첨가하였고, 이어서 LiI(280㎎, 4.12m㏖) 및 DIPEA(0.41㎖, 2.47m㏖)을 첨가하였다. 얻어진 혼합물을 약 22℃에서 18시간 교반한 다음, 냉각 EtOAc(20㎖) 및 냉각 H₂O(20㎖)로서 희석하였고 셀라이드 패드를 통과시켜 고체를 제거하였다. 패드를 EtOAc(2% 20㎖)로 세척하였고 두개의 용액상을 분리하였다. 수성상을 EtOAc(1 × 20㎖)로 추출하였고, 유기 분율을 결합하고, H₂O(3 × 25㎖) 및 염수 (7 × 25㎖)로 세척하고, 건조시킨다음 (MgSO₄) 농축시켰다. 잔류물을 섬광 실리카 겔 컬럼(실리카 20g; 컬럼을 처음에 CH₂Cl₂로서 이어서 연속적으로 5%, 10%, 15% 및 20% EtOAc/CH₂Cl₂로서 용출함)을 통과시켜 표제의 물질(462㎎, 40%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3- is mercapto-4-methyl-7-oxo-1-azabicyclo in DMF (6 mL) A newly prepared 1-chloromethylthio-3,4-dichlorobenzene solution in a cooled (ice bath) solution of [3.2.0] heptan-2-ene-2-carboxylate (1.00 g, 2.06 mmol) ( 1.27 mL of 3,4-dichlorophenyl mercaptan made from 10.0 mmol) was added followed by LiI (280 mg, 4.12 mmol) and DIPEA (0.41 mL, 2.47 mmol). The resulting mixture was stirred at ˜22 ° C. for 18 hours, then diluted with cold EtOAc (20 mL) and cold H ₂ O (20 mL) and passed through a pad of celide to remove solids. The pad was washed with EtOAc (2% 20 mL) and the two solution phases were separated. The aqueous phase was extracted with EtOAc (1 × 20 mL), organic fractions combined, washed with H ₂ O (3 × 25 mL) and brine (7 × 25 mL), dried and concentrated (MgSO ₄ ). . The residue was passed through a flash silica gel column (20 g of silica; eluting the column initially as CH ₂ Cl ₂ and subsequently as 5%, 10%, 15% and 20% EtOAc / CH ₂ Cl ₂ ) to obtain the title material. (462 mg, 40%).

[단계 B][Step B]

소듐 (4R,5S,6S)-3-[[[(3,4-디클로로페닐)티오]메틸]티오-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iq)Sodium (4R, 5S, 6S) -3-[[[(3,4-dichlorophenyl) thio] methyl] thio-6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo- 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iq)

THF(15㎖), 에테르(15㎖) 및 0.10M pH 7.0 NaH₂PO₄/NaOH 완충용액 (14.5㎖, 1.45m㏖)내 p-니트로벤질 (4R,5S,6S)-3-[[[(3,4-디클로로페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(447㎎, 0.786m㏖) 용액을 촉매로서 10% Pd/C(447㎎)을 사용하여 40-45 psi H₂에서 2시간 동안 Parr 수소첨가 장치에서 교반하였다. 여과에 의하여 촉매를 제거하였고, 완충용액(2 × 10㎖)으로 세척하였다. 두개의 용액상을 분리하였고 유기상을 완충용액 (2 × 10㎖)으로 추출하였다. 수성상을 결합하고, 에테르(3×20㎖)로 세척한 다음 C₁₈μBondaPak 역상 컬럼으로 2회 통과시켜(제1용출: C₁₈μBondaPak 역상물질 27g; 처음에 H₂O로서 이어서 연속적으로 2%, 5%, 10% 및 20% CH₃CN/H₂O로 용출함; 제2용출; 역상물질 7.5g;H₂O 및 5%, 10%와 20% CH₃CN/H₂O로서 연속적으로 용출함) 친액화 분말로서 표제의 물질(102㎎, 29%)을 제공하였다: T¹/₂= 99시간 (pH 7.4, 37℃), 순도 99.3% (HPLC에 의하여 검사).P-nitrobenzyl (4R, 5S, 6S) -3-[[[[] in 0.15M pH 7.0 NaH ₂ PO ₄ / NaOH buffer (14.5 mL, 1.45 mmol), THF (15 mL), ether (15 mL). (3,4-dichlorophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2- En-2-carboxylate (447 mg, 0.786 mmol) solution was stirred in a Parr hydrogenator for 2 hours at 40-45 psi H ₂ using 10% Pd / C (447 mg) as catalyst. The catalyst was removed by filtration and washed with buffer (2 × 10 mL). The two solution phases were separated and the organic phase was extracted with buffer (2 × 10 mL). The aqueous phases were combined, washed with ether (3 × 20 mL) and then passed twice through a C ₁₈ μBondaPak reversed phase column (first elution: 27 g of C ₁₈ μBondaPak reversed phase; initially as H ₂ O followed by 2% consecutively Eluted with 5%, 10% and 20% CH ₃ CN / H ₂ O; second eluted; 7.5 g of reverse phase; H ₂ O and 5%, 10% and 20% CH ₃ CN / H ₂ O elution hereinafter) of the title substance as a parent liquefied powder (102㎎, to give a 29%) to: check by T ^_1/2 = 99 sigan (pH 7.4, 37 ℃), purity 99.3% (HPLC).

[실시예 20]Example 20

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(2,3,4,5,6-펜타플루오로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Ir)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(2,3,4,5,6-pentafluorophenyl) thio] Methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Ir)

[단계A][Step A]

p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(2,3,4,5,6-펜타플루오로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[((2,3,4,5,6-pentafluorophenyl) ) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF(12㎖) 내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(2.0g, 4.12m㏖) 냉각(얼음조) 용액을 새로 제조된 DMF(18㎖)내 1-클로로메틸티오-2,3,4,5,6-펜타플루오로벤젠용액(1-메르캅토-2,3,4,5,6-펜타플루오로벤젠; 1.33㎖, 100m㏖에서 제조됨), LiI(560㎎, 8.24m㏖) 및 DIPEA(0.82㎖, 4.94m㏖)로 처리하였다. 혼합물을 약22℃에서 18시간 교반한 다음, 얼음냉각 H₂O(40㎖) 및 EtOAc(40㎖)로 희석하였고 셀라이트 패드위에서 여과시켰다. 패드를 EtOAc(3 × 10㎖)로 린스하였고 두개의 용액상을 분리하였다. 수성층을 EtOAc (2 × 20㎖)로 추출하였다. 그후 유기상을 결합하였고, 얼음냉각 H₂O (3 ×50㎖) 및 염수(50㎖)로 세척하였고, 건조 시킨다음(MgSO₄) 농축시켰다. 잔류물을 실리카 겔 컬럼으로 통과시켜 (실리카 40g; 컬럼을 CH₂Cl₂및 2%, 5%, 8% 및 10% EtOAc/CH₂Cl₂로서 연속 용출함) 표제의 물질(820㎎, 34%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1-azabicyclo in DMF (12 mL) [3.2.0] Heptane-2-ene-2-carboxylate (2.0 g, 4.12 mmol) cooled (ice bath) solution was prepared in 1-chloromethylthio-2,3, in freshly prepared DMF (18 mL). 4,5,6-pentafluorobenzene solution (1-mercapto-2,3,4,5,6-pentafluorobenzene; 1.33 mL, prepared in 100 mmol), LiI (560 mg, 8.24 mmol) ) And DIPEA (0.82 mL, 4.94 mmol). The mixture was stirred at about 22 ° C. for 18 hours, then diluted with ice cold H ₂ O (40 mL) and EtOAc (40 mL) and filtered over a pad of celite. The pad was rinsed with EtOAc (3 × 10 mL) and the two solution phases were separated. The aqueous layer was extracted with EtOAc (2 × 20 mL). The organic phases were then combined, washed with ice cold H ₂ O (3 × 50 mL) and brine (50 mL), dried (MgSO ₄ ) and concentrated. The residue was passed through a silica gel column (silica 40 g; the column was eluted continuously with CH ₂ Cl ₂ and 2%, 5%, 8% and 10% EtOAc / CH ₂ Cl ₂ ) titled material (820 mg, 34 %).

[단계 B][Step B]

THF(10㎖), 에테르(10㎖) 및 0.10M pH 7.0 NaH₂PO₄/NaOH 완충용액(6.3㎖, 0.63m㏖) 내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(2,3,4,5,6-펜타플루오로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(200㎎, 0.34m㏖) 용액을 촉매로서 10% Pd/C(200㎎)을 사용하여 40-45 psi H₂에서 3시간 동안 파르 수소첨가장치에서 교반하였다. 여과에 의하여 촉매를 제거하였고, pH 7.0 완충용액 (2 × 5㎖) 및 에테르 (1 × 10㎖)로서 린스하였다. 두개의 용액층을 분리하였고, 유기층을 pH 7.0 완충용액(2 × 5㎖)으로 추출하였다. 추가 조작이 필요할 때까지 수성상을 동결시켰다. 유기층을 다시 파르교반기에서 40-45 psi H₂에서 2시간 동안 10% Pd/C 촉매(130㎎)로서 처리하였고 상기에 기재된 유사한 추출공정을 수행하였다. 끝으로, 모든 수성층을 결합하였고, C₁₈μBondaPak 역상 컬럼으로 통과시켜 (C₁₈μBondaPak 역상물질 30g; 컬럼을 처음에 H₂O로서 이어서 2%, 5% 및 10% CH₃CN/H₂O 추출함)로 표제의 물질(45㎎, 28%)을 제공하였다: 순도 100% (HPLC로 검사); T¹/₂= 41시간 (pH 7.4, 37℃).P-nitrobenzyl (4R, 5S, 6S) -6- [1 'in THF (10 mL), ether (10 mL) and 0.10 M pH 7.0 NaH ₂ PO ₄ / NaOH buffer (6.3 mL, 0.63 mmol) (R) -hydroxyethyl] -4-methyl-3-[[[((2,3,4,5,6-pentafluorophenyl) thio] methyl] thio] -7-oxo-1-azabicyclo [ 3.2.0] Heptane solution of heptane-2-ene-2-carboxylate (200 mg, 0.34 mmol) for 10 hours at 40-45 psi H ₂ using 10% Pd / C (200 mg) as catalyst Stir in a hydrogenator. The catalyst was removed by filtration and rinsed with pH 7.0 buffer (2 × 5 mL) and ether (1 × 10 mL). The two solution layers were separated and the organic layer was extracted with pH 7.0 buffer (2 x 5 mL). The aqueous phase was frozen until further manipulation was needed. The organic layer was again treated with 10% Pd / C catalyst (130 mg) for 2 hours at 40-45 psi H ₂ in a Parrizer and the similar extraction process described above was carried out. Finally, all aqueous layers were combined and passed through a C ₁₈ μBondaPak reversed phase column (30 g of C ₁₈ μBondaPak reversed phase; the column was initially extracted with H ₂ O followed by 2%, 5% and 10% CH ₃ CN / H ₂ O ) Provided the title material (45 mg, 28%): purity 100% (check by HPLC); T ^_1/2 = 41 sigan (pH 7.4, 37 ℃).

[실시예 21]Example 21

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[(이소프로필티오)메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Is)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(isopropylthio) methyl] thio] -4-methyl-7-oxo-1-azabicyclo [ 3.2.0] heptan-2-ene-2-carboxylate (Is)

[단계A][Step A]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[(이소프로필티오)메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(isopropylthio) methyl] thio] -4-methyl-7-oxo-1- Azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

건조 디메틸포름아미드 60㎖내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(7.279g, 15.0m㏖) 냉각(2℃)용액을 건조 디메틸포름아미드 15㎖내 2-[(클로로메틸)티오]프로판(2.805g, 22.5m㏖)용액, 요오드화 리튬(6.023g, 45.0m㏖) 및 N,N-디이소프로필에틸아민(2.908g, 3.92㎖, 22.5m㏖)으로서 처리하였다. 5℃에서 1시간 및 20℃에서 18시간 교반후, 용액을 초산 에틸(150㎖) 및 냉각수 (2℃)(150㎖)로 희석한 다음 셀라이트 패드를 통하여 여과시켰다. 유기상을 수성상으로 부터 분리하였고, 수성상을 초산 에틸(3 × 150㎖)로서 추출하였다. 유기층을 결합하고, 물(3 × 100㎖) 및 염수(200㎖)로 세척하고, 건조시키고(MgSO₄), 여과시킨다음 증발시켰다. 원생성물(4.87g)을 실리카 겔 크로마토그래피(실리카 250g; 디클로로메탄/초산 에틸, 3/1로서 용출)에 의하여 정제시켜 황색 발포체로서 표제의 생성물 2.483g(35%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- silver mercapto-7-oxo-1-azabi in 60 ml of dry dimethylformamide Cyclo [3.2.0] heptane-2-ene-2-carboxylate (7.279 g, 15.0 mmol) The solution was cooled (2 ° C.) to 2-[(chloromethyl) thio] propane (in 15 ml of dry dimethylformamide). 2.805 g, 22.5 mmol) solution, lithium iodide (6.023 g, 45.0 mmol) and N, N-diisopropylethylamine (2.908 g, 3.92 mL, 22.5 mmol) were treated. After stirring for 1 hour at 5 ° C. and 18 hours at 20 ° C., the solution was diluted with ethyl acetate (150 mL) and cooling water (2 ° C.) (150 mL) and then filtered through a pad of celite. The organic phase was separated from the aqueous phase and the aqueous phase was extracted as ethyl acetate (3 x 150 mL). The organic layers were combined, washed with water (3 × 100 mL) and brine (200 mL), dried (MgSO ₄ ), filtered and evaporated. The crude product (4.87 g) was purified by silica gel chromatography (250 g silica; eluted as dichloromethane / ethyl acetate, 3/1) to give 2.483 g (35%) of the title product as a yellow foam.

[단계B][Step B]

에테르(30㎖) 및 테트라히드로푸란(30㎖)의 혼합물 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[(이소프로필티오)메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(1.399g, 3.0m㏖) 용액을 pH 7.0, 0.1M NaH₂PO₄/NaOH 완충액 60 ㎖에 첨가하였다. 얻어진 혼합물을 42 psi 수소에서 3시간 동안 10% Pd/C 촉매(1.399g)위에서 가수소분해반응시켰다. 셀라이트 패드위에서 여과에 의하여 촉매를 제거하였고 에테르(30㎖) 및 pH 7.0 완충용액(30㎖)으로 세척하였다. 수성상을 분리한 다음 물내 5-15% 아세토니트릴로니 용출된 역상 실리카 겔에서 크로마토그래피하였다. 관련 분율을 모아 친액화하였다. 얻어진 고체를 아세토니트릴/물(12/88)로서 용출된 역상 실리카 겔에서 재크로마토그래피하였다. 다시한번, 관련 분율을 모았고, 친액화하여 약간 베이지색 고체로서 표제의 생성물 0.357g(33%)을 제공하였다.Mixture of ether (30 mL) and tetrahydrofuran (30 mL) p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(isopropylthio) Methyl] thio] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (1.399 g, 3.0 mmol) solution in pH 7.0, 0.1 M NaH ₂ To 60 ml PO ₄ / NaOH buffer was added. The resulting mixture was hydrolyzed on 10% Pd / C catalyst (1.399 g) in 42 psi hydrogen for 3 hours. The catalyst was removed by filtration over a pad of celite and washed with ether (30 mL) and pH 7.0 buffer (30 mL). The aqueous phase was separated and then chromatographed on reversed phase silica gel eluted with 5-15% acetonitrile in water. Relevant fractions were collected and lyophilised. The solid obtained was rechromatated on reversed phase silica gel eluted as acetonitrile / water (12/88). Once again the relevant fractions were collected and lysed to give 0.357 g (33%) of the title product as a slightly beige solid.

[실시예 22]Example 22

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[(페닐메틸)티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(It)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(phenylmethyl) thio] methyl] thio] -4-methyl-7-oxo-1-azabi Cyclo [3.2.0] heptan-2-ene-2-carboxylate (It)

[단계A][Step A]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[(페닐메틸)티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(phenylmethyl) thio] methyl] thio] -4-methyl-7-oxo- 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

건조 디메틸포름아미드 60㎖내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(7.279g, 15.0m㏖) 냉각(2℃)용액을 건조 디메틸포름아미드 15㎖내 클로로메틸티오메틸벤젠 (3.885g, 22.5m㏖) 용액, 요오드화 리튬 (6.023g, 45.0m㏖) 및 N,N-디이소프로필에틸아민 (2.908g, 3.92㎖, 22.5m㏖)으로서 처리하였다. 5℃에서 1시간 및 20℃에서 18시간 교반후, 용액을 초산 에틸(150㎖) 및 냉각수 (2℃)(150㎖)로 희석한 다음 셀라이트 패드를 통하여 여과시켰다. 유기상을 수성상으로 부터 분리하였고, 수성상을 초산 에틸(3 × 150㎖)로서 추출하였다. 유기상을 결합하고, 물(3 × 100㎖) 및 염수(200㎖)로 세척하였고, 건조시킨 다음(MgSO₄), 여과 증발시켰다. 원생성물(5.18g)을 실리카 겔 크로마토그래피(실리카 250g; 디클로로메탄/초산 에틸, 3/1로서 용출)에 의하여 정제시켜 황색 발포체로서 표제의 생성물 2.772g(36%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- silver mercapto-7-oxo-1-azabi in 60 ml of dry dimethylformamide Cyclo [3.2.0] heptane-2-ene-2-carboxylate (7.279 g, 15.0 mmol) Cooled (2 ° C.) solution to 15 ml of dimethylformamide in chloromethylthiomethylbenzene (3.885 g, 22.5 m Mol) solution, lithium iodide (6.023 g, 45.0 mmol) and N, N-diisopropylethylamine (2.908 g, 3.92 mL, 22.5 mmol). After stirring for 1 hour at 5 ° C. and 18 hours at 20 ° C., the solution was diluted with ethyl acetate (150 mL) and cooling water (2 ° C.) (150 mL) and then filtered through a pad of celite. The organic phase was separated from the aqueous phase and the aqueous phase was extracted as ethyl acetate (3 x 150 mL). The organic phases were combined, washed with water (3 × 100 mL) and brine (200 mL), dried (MgSO ₄ ) and filtered evaporated. The crude product (5.18 g) was purified by silica gel chromatography (250 g silica; eluted as dichloromethane / ethyl acetate, 3/1) to give 2.772 g (36%) of the title product as a yellow foam.

[단계B][Step B]

에테르(30㎖) 및 테트라히드로푸란(30㎖)의 혼합물내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[(페닐메틸)티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(1.544g, 3.0m㏖) 용액을 pH 7.0, 0.1M NaH₂PO₄/NaOH 완충용액에 첨가하였다. 얻어진 혼합물을 10% Pd/C 촉매(1.544g)위에서 42 psi H₂에 3시간 동안 가수소분해반응시켰다. 셀라이트 패드위에서 여과에 의하여 촉매를 제거하였고 에테르(30㎖) 및 pH 7.0 완충용액(30㎖)으로 세척하였다. 수성상을 유기상으로 부터 분리하였고, 물내 5-20% 아세토니트릴로 용출된 역상 실리카 겔에서 크로마토그래피하였고; 관련 분율을 모아 친액화하였다. 따라서 얻어진 고체를 아세토니트릴/물(15/85)로서 용출된 역상 실리카 겔에서 재크로마토그래피하였고; 관련 분율을 다시한번 모아 친액화하여 백색분말로서 표제의 화합물 0.390g(32%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(phenylmethyl in a mixture of ether (30 mL) and tetrahydrofuran (30 mL) ) Thio] methyl] thio] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (1.544 g, 3.0 mmol) in a solution of pH 7.0, 0.1 It was added to M NaH ₂ PO ₄ / NaOH buffer. The resulting mixture was hydrolyzed at 42 psi H ₂ for 3 hours on 10% Pd / C catalyst (1.544 g). The catalyst was removed by filtration over a pad of celite and washed with ether (30 mL) and pH 7.0 buffer (30 mL). The aqueous phase was separated from the organic phase and chromatographed on reverse phase silica gel eluted with 5-20% acetonitrile in water; Relevant fractions were collected and lysified. The solid thus obtained was rechromatography on reversed phase silica gel eluted as acetonitrile / water (15/85); The relevant fractions were collected once again and lyophilised to give 0.390 g (32%) of the title compound as a white powder.

[실시예 23]Example 23

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[[((푸란-2-일)-메틸]티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iu)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[[((furan-2-yl) -methyl] thio] methyl] thio] -4-methyl -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iu)

[단계A][Step A]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[[((푸란-2-일)메틸]티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[[((furan-2-yl) methyl] thio] methyl] thio] -4 -Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

건조 디메틸포름아미드 60㎖내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸--3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(7.279g, 15.0m㏖) 냉각(2℃)용액을 건조 디메틸포름아미드 15㎖내 2-[[(클로로메틸)티오]메틸]푸란 (3.659g, 22.5m㏖) 용액, 요오드화 리튬(6.023g, 45.0m㏖) 및 N,N-디이소프로필에틸아민(2.908g, 3.92㎖, 22.5m㏖)으로서 처리하였다. 5℃에서 1시간 및 20℃에서 18시간 교반후, 용액을 초산 에틸(150㎖) 및 냉각수 (2℃)(150㎖)로 희석하였고 셀라이트 패드 위에서 여과시켰다. 수성상으로부터 유기상을 분리하였고; 수성상을 초산 에틸(3 × 150㎖)로서 추출하였다. 유기상을 결합하였고, 물(3 × 100㎖) 및 염수(200㎖)로 세척하였고, 건조시킨 다음(MgSO₄), 여과 및 증발시켰다. 원생성물(3.80g)을 실리카 겔 크로마토그래피(실리카 250g; 디클로로메탄/초산 에틸, 3/1로서 용출함)에 의하여 정제시켜 황색 발포체로서 표제의 생성물 2.471g(33%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl--3- is mercapto-7-oxo-1- in 60 ml of dry dimethylformamide Azabicyclo [3.2.0] heptane-2-ene-2-carboxylate (7.279 g, 15.0 mmol) Cooled (2 ° C.) solution was dried in 15 ml of dimethylformamide in 2-[[(chloromethyl) thio]. Treated with methyl] furan (3.659 g, 22.5 mmol) solution, lithium iodide (6.023 g, 45.0 mmol) and N, N-diisopropylethylamine (2.908 g, 3.92 mL, 22.5 mmol). After stirring for 1 hour at 5 ° C. and 18 hours at 20 ° C., the solution was diluted with ethyl acetate (150 mL) and cooling water (2 ° C.) (150 mL) and filtered over a pad of celite. The organic phase was separated from the aqueous phase; The aqueous phase was extracted as ethyl acetate (3 x 150 mL). The organic phases were combined, washed with water (3 × 100 mL) and brine (200 mL), dried (MgSO ₄ ), filtered and evaporated. The crude product (3.80 g) was purified by silica gel chromatography (250 g silica; eluted as dichloromethane / ethyl acetate, 3/1) to give 2.471 g (33%) of the title product as a yellow foam.

[단계B][Step B]

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[[((푸란-2-일)메틸]티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iu)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[[((furan-2-yl) methyl] thio] methyl] thio] -4-methyl- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iu)

에테르(30㎖) 및 테트라히드로푸란(30㎖)의 혼합물내 p-니트로벤질(4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-[[[[((푸란-2-일)메틸]티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (1.512g, 3.0m㏖) 용액을 pH 7.0, 0.1M NaH₂PO₄/NaOH 완충용액 60 ㎖에 첨가하였다. 얻어진 혼합물을 10% Pd/C 촉매 (1.512g)위에서 42 psi H₂에서 3시간 동안 가수소분해반응시켰다. 셀라이트 패드에서 여과에 의하여 촉매를 제거하였고 에테르(30㎖) 및 pH 7.0 완충용액(30㎖)으로 세척하였다. 유기상으로 부터 수성상을 분리하였고 물내 5-20% 아세토니트릴로 용출된 역상 실리카 겔에서 크로마토그래피하였고; 관련 분율을 모아 친액화하였다. 이와 같이 얻어진 고체를 아세토니트릴/물(12/88)로서 용출된 역상 실리카 겔에서 재크로마토그래피하였고; 관련 분율을 다시한번 모아 친액화시켜 백색 분말로서 표제생성물 0.339g(29%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[[(((2) in a mixture of ether (30 mL) and tetrahydrofuran (30 mL) Furan-2-yl) methyl] thio] methyl] thio] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (1.512 g, 3.0 mmol ) Solution was added to 60 ml of 0.1 M NaH ₂ PO ₄ / NaOH buffer, pH 7.0 The resulting mixture was hydrolyzed at 42 psi H ₂ for 3 hours on 10% Pd / C catalyst (1.512 g). The catalyst was removed by filtration in a pad of celite and washed with ether (30 mL) and pH 7.0 buffer (30 mL) The aqueous phase was separated from the organic phase and reversed phase silica gel eluted with 5-20% acetonitrile in water. The resulting fractions were lyophilised and the resulting solid was rechromatated on reversed phase silica gel eluted as acetonitrile / water (12/88); Liquiphilization gave 0.339 g (29%) of the title product as a white powder.

[실시예 24]Example 24

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(1(R 및 S)-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iv)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(1 (R and S)-(methylthio) ethyl] thio] -7- Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iv)

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(1-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(1- (methylthio) ethyl] thio] -7-oxo- 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF(2㎖)중 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (242㎎, 0.5 m㏖)의 냉각(얼음조)용액을 DMF(1㎖)내 1-클로로에틸 메틸 술파이드 (69㎎, 0.625m㏖) 용액, LiI (68㎎, 1.0 m㏖) 및 DIPEA(0.1㎖, 0.6m㏖)로서 적가 처리하였다. 얻어진 혼합물을 5℃에서 30분간 교반한 다음, 다시 이전의 시약(CH₃CHClSCH₃, LiI 및 DIPEA)0.5당량으로 처리하였다. 혼합물을 추가의 30분간 교반한 다음, 얼음 냉각 EtOAc(20㎖) 및 H₂O(20㎖)로 희석하였고 셀라이드 패드에서 여과하였다. 패드를 EtOAc(3 × 10㎖)로 린스하였고 두개의 용액층을 분리하였다. 수성층을 EtOAc(3 × 10㎖)로 추출하였다. 유기상을 결합하고 얼음 냉각 H₂O(4 × 20㎖) 및 염수(20㎖)로 세척하고, 건조시킨다음(MgSO₄), 농축시켰다. 잔류물을 예비 TLC(CH₂Cl₂/EtOAc 1/1로 용출)에서 정제시켜 부분입체이성체의 혼합물로서 표제 화합물(73㎎, 32%)을 제공하였다. 각 부분입체이성체를 예비 TLC(에테르로 용출)에 의하여 분리하였고 별도로 특성화하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1-azabicyclo in DMF (2 mL) [3.2.0] Chilled (ice bath) solution of heptane-2-ene-2-carboxylate (242 mg, 0.5 mmol) was diluted with 1-chloroethyl methyl sulfide (69 mg, 0.625) in DMF (1 mL). mmol) solution, LiI (68 mg, 1.0 mmol) and DIPEA (0.1 mL, 0.6 mmol) were added dropwise. The resulting mixture was stirred at 5 ° C. for 30 min and then again treated with 0.5 equivalents of the previous reagents (CH ₃ CHClSCH ₃ , LiI and DIPEA). The mixture was stirred for an additional 30 minutes, then diluted with ice cold EtOAc (20 mL) and H ₂ O (20 mL) and filtered over a pad of celide. The pad was rinsed with EtOAc (3 × 10 mL) and the two solution layers were separated. The aqueous layer was extracted with EtOAc (3 × 10 mL). The organic phases were combined and washed with ice cold H ₂ O (4 × 20 mL) and brine (20 mL), dried (MgSO ₄ ) and concentrated. The residue was purified on preparative TLC (eluted with CH ₂ Cl ₂ / EtOAc 1/1) to give the title compound (73 mg, 32%) as a mixture of diastereomers. Each diastereomer was separated by preparative TLC (eluted with ether) and characterized separately.

[이성체A, 극성이 적은 이성체][Isomer A, Isomer with Less Polarity]

[이성체B, 극성이 많은 이성체][Isomer B, Isomer with High Polarity]

[단계B][Step B]

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(1-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iv)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(1- (methylthio) ethyl] thio] -7-oxo-1-azabi Cyclo [3.2.0] heptan-2-ene-2-carboxylate (Iv)

THF(10㎖), 에테르(10㎖) 및 0.10M NaH₂PO₄/NaOH 완충용액(4.7㎖, 0.47 m㏖) 내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(1-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(이성체A, 120㎎, 0.26m㏖)용액을 파르 수소 첨가장치에서 촉매로서 10% Pd/C (120㎎)을 사용하여 40-45 psi H₂에 1.5시간 교반하였다. 여과에 의하여 촉매를 제거하였고 수성상으로부터 유기상을 분리하였고 10% Pd/C 촉매(120㎎) 및 pH 7.0 NaH₂PO₄/NaOH 완충용액 존재하에 파르 교반기에서 40-45 psi에서 H₂로서 다시 처리하였다. 촉매를 여과에 의하여 제거한 후 혼합물을 1.5시간 동안 교반하였다. 두개의 가수소분해단계로 부터 수성상을 결합하였고, 에테르(3 × 20㎖)로 세척한 다음 C₁₈μBondaPak 컬럼(C₁₈μBondaPak 물질 30g; 컬럼을 처음에 H₂O로 이어서 연속적으로 2%, 5% 및 10% CH₃CN/H₂O로 용출함)으로 통과시켜 회색 고체로서 표제의 물질을 제공하였다. 고체를 C₁₈μBondaPak 컬럼(C₁₈μBondaPak 물질 7g; 연속적으로 H₂O, 5% 및 10% CH₃CN/H₂O로 용출함)에서 재정제하여 한가지 순수 이성체 Iv-A(35㎎, 40%)를 제공하였다: 순도 96.9% (HPLC로 검사); T¹/₂20분 (pH2, 37℃).P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) in THF (10 mL), ether (10 mL) and 0.10 M NaH ₂ PO ₄ / NaOH buffer (4.7 mL, 0.47 mmol). ) -Hydroxyethyl] -4-methyl-3-[[(1- (methylthio) ethyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxyl The rate (isomer A, 120 mg, 0.26 mmol) solution was stirred for 1.5 h in 40-45 psi H ₂ using 10% Pd / C (120 mg) as a catalyst in a Parr hydrogenation apparatus. The organic phase was separated from the aqueous phase and treated again as H ₂ at 40-45 psi in a Parr stirrer in the presence of 10% Pd / C catalyst (120 mg) and pH 7.0 NaH ₂ PO ₄ / NaOH buffer. The mixture was stirred for 1.5 h after being removed by immersion of the aqueous phases from the two hydrolysis steps, washed with ether (3 × 20 mL) and then washed with C ₁₈ μBondaPak column (30 g of C ₁₈ μBondaPak material; column the first with H ₂ O followed successively 2%, 5% . By passing a 10% CH ₃ eluting with CN / H ₂ O) to give a material of the title as a white solid The solid C ₁₈ μBondaPak column (C ₁₈ μBondaPak material 7g; subsequently H ₂ O, 5% and 10% CH ₃ and provided eluting with CN / H ₂ O) to override the one kinds of pure isomer Iv-a (35㎎, 40% ) in: testing a purity of ^{96.9% (HPLC); T 1} /2 20 bun (pH2, 37 ° C.).

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(1-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트의 이성체B로서 유사한 단계는 또다른 이성체 Iv-B를 제공하였다(37㎎, 42%): 순도 99.0%(HPLC로 검사); T¹/₂56시간 (pH 7.4, 37℃), 20분 (pH 2.0, 37℃)p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(1- (methylthio) ethyl] thio] -7-oxo- Similar step as isomer B of 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate gave another isomer Iv-B (37 mg, 42%): purity 99.0% (by HPLC) inspection); T ^_1/2 56 sigan (pH 7.4, 37 ℃), 20 bun (pH 2.0, 37 ℃)

[실시예 25]Example 25

소듐 (4R,5S,6S)-3-[[[(p-시아노페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(Iw)Sodium (4R, 5S, 6S) -3-[[[(p-cyanophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo- 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iw)

[단계A][Step A]

4-[(브로모메틸)티오] 벤조니트릴4-[(bromomethyl) thio] benzonitrile

벤젠내 4-(메틸티오)벤조니트릴(0.300g, 2m㏖) 및 N-브로모숙신이미드(0.445g, 2.5m㏖) 용액을 26시간동안 환류시켰다. 그후 혼합물을 냉각시킨다음 여과시켰다. 여과물을 증발시키고, 냉각 CCl₄(10㎖)에서 취한 다음 다시 여과시켰다. 이와 같은 공정을 다시 한번 반복하였다. 끝으로, 용매의 증발은 고체화된 황색 오일로서 표제 생성물 0.32g(70%)을 남겼다.A solution of 4- (methylthio) benzonitrile (0.300 g, 2 mmol) and N-bromosuccinimide (0.445 g, 2.5 mmol) in benzene was refluxed for 26 hours. The mixture was then cooled and filtered. The filtrate was evaporated, taken up in cold CCl ₄ (10 mL) and filtered again. This process was repeated once again. Finally, evaporation of the solvent left 0.32 g (70%) of the title product as a solidified yellow oil.

[단계B][Step B]

p-니트로벤질 (4R,5S,6S)-3-[[[(p-시아노페닐)티오]메틸]티오]-6- [1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[(p-cyanophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

디메틸포름아미드 (5㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(0.606g, 1.25m㏖) 냉각(5℃)용액에 디메틸포름아미드 (1㎖)내 4-[(브로모메틸)티오]벤조니트릴(0.328g, 1.44m㏖)용액, 이어서 LiI(0.255g, 3.75m㏖) 및 N,N-디이소프로필에틸아민 (0.37㎖, 2.17m㏖)을 적가하였다. 반응 혼합물을 상온에서 18시간 교반한 다음, 초산 에틸(25㎖) 및 물(25㎖)로 희석하였고 여과시켰다. 두 개의 층을 분리하였고, 수성상을 초산 에틸(3 × 10 ㎖)로 추출하였다. 유기상을 결합하였고 냉각수 및 염수로 세척하였다. 그후 유기용액을 건조시킨 다음(MgSO₄) 증발시켰다. 원화합물을 실리카 겔 크로마토그래피(0% - 20% 초산 에틸/CH₂Cl₂로 용출)에 의하여 정제시켜 황색 고체로서 표제 생성물 0.298g(45%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1- in dimethylformamide (5 mL) 4-[(bromomethyl) thio in dimethylformamide (1 mL) in azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.606 g, 1.25 mmol) cooling (5 ° C) solution ] Benzonitrile (0.328 g, 1.44 mmol) solution, followed by LiI (0.255 g, 3.75 mmol) and N, N-diisopropylethylamine (0.37 mL, 2.17 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 18 hours, then diluted with ethyl acetate (25 mL) and water (25 mL) and filtered. The two layers were separated and the aqueous phase was extracted with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with cool water and brine. The organic solution was then dried (MgSO ₄ ) and evaporated. The crude compound was purified by silica gel chromatography (eluted with 0% -20% ethyl acetate / CH ₂ Cl ₂ ) to give 0.298 g (45%) of the title product as a yellow solid.

[단계C][Step C]

테트라히드로푸란(12㎖)내 p-니트로벤질 (4R,5S,6S)-3-[[[(p-시아노페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트(0.168g, 0.32m㏖) 용액을 Et₂O(12㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액(pH7.0, 6㎖)의 혼합물에 첨가하였다. 혼합물을 10% Pd/C 촉매(0.168g)위에서 45 psi H₂에 2.75시간동안 가수소분해반응 시켰다. 그후, 촉매를 여과시키고 여과물을 Et₂O로 추출하였다. 수성사을 0-10% CH₃CN/물로서 용출된 역상 실리카 겔에서 크로마토그래피하였다. 관련 분율을 결합하고 친액화하여 표제 화합물 0.30g(22.7%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -3-[[[(p-cyanophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxy in tetrahydrofuran (12 mL) Ethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.168 g, 0.32 mmol) was dissolved in Et ₂ O (12 mL) and 0.1 To a mixture of M NaH ₂ PO ₄ / NaOH buffer (pH 7.0, 6 mL) was added. The mixture was hydrolyzed in 45 psi H ₂ for 2.75 hours on 10% Pd / C catalyst (0.168 g). The catalyst was then filtered off and the filtrate was extracted with Et ₂ O. The aqueous sand was chromatographed on reversed phase silica gel eluted as 0-10% CH ₃ CN / water. The relevant fractions were combined and lyophilised to give 0.30 g (22.7%) of the title compound.

[실시예 26]Example 26

소듐 (4R,5S,6S)-3-[[[(p-카르바모일페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Ix)Sodium (4R, 5S, 6S) -3-[[[[(p-carbamoylphenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo -1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Ix)

[단계 A][Step A]

4-[(클로로메틸)티오]벤즈아미드4-[(chloromethyl) thio] benzamide

SOCl₂(5㎖) 내 4-(메틸티오)벤조산 (1.68g, 10m㏖) 용액을 1시간 동안 환류시켰다. 그후 용매를 증발시켜 고체를 남겼다. 형성된 원 산염화물을 CH₂Cl₂(10㎖)에 용해시켰고, 얼음조에서 냉각시킨다음 CH₂Cl₂(5㎖)내 SO₂Cl₂(1.48g, 11m㏖) 용액으로 천천히 (45분) 처리하였다. 혼합물을 5℃에서 2시간동안 교반한후, 용매를 증발시켰다. 원 생성물을 C₆H₆(50㎖)에 용해시켰고 용액을 20℃ 에서 NH₃로서 포화시켰다. 즉시 침전물이 형성되었다. 15분간 교반시킨후, 백색 고체를 수집하였고 실리카겔 (CH₃CN 으로 용출)에서 크로마토그래피하여 표제의 아미드 1.74g (86.2%)을 제공하였다 : m.p. 140-142℃.A solution of 4- (methylthio) benzoic acid (1.68 g, 10 mmol) in SOCl ₂ (5 mL) was refluxed for 1 hour. The solvent was then evaporated to leave a solid. The crude acid chloride formed was dissolved in CH ₂ Cl ₂ (10 mL), cooled in an ice bath and treated slowly with a solution of SO ₂ Cl ₂ (1.48 g, 11 mmol) in CH ₂ Cl ₂ (5 mL) (45 min). It was. The mixture was stirred at 5 ° C. for 2 hours and then the solvent was evaporated. The crude product was dissolved in C ₆ H ₆ (50 mL) and the solution was saturated as NH ₃ at 20 ° C. A precipitate formed immediately. After stirring for 15 minutes, a white solid was collected and chromatographed on silica gel (eluted with CH ₃ CN) to give 1.74 g (86.2%) of the title amide: mp 140-142 ° C.

[단계 B][Step B]

p-니트로벤질 (4R,5S,6S)-3-[[[(p-카르바모일페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[(p-carbamoylphenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

디메틸포름아미드 (10㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (500㎎, 1.03m㏖, 냉각 (5℃) 용액을 4-[(클로로메틸)티오]벤즈아미드 (0.239g, 1.19m㏖), LiI (0.414g, 3.09m㏖) 및 N,N-디이소프로필에틸아민 (0.233g, 1.8m㏖)을 적가처리하였다. 혼합물을 5℃ 에서 2시간 및 20℃ 에서 16시간 교반하였고, 초산에틸로 희석하고, 냉각된 묽은 HCl로서 세게 교반한다음 여과시켰다. 두개의 용액상을 분리하였고, 수성용액을 초산에틸로서 추출하였다. 결합된 유기추출물을 염수로 세척하였고, 건조시킨다음 (MgSO₄) 증발시켰다. 원 생성물을 실리카겔 크로마토그래피 (처음에 초산에틸로 그 후 CH₃CN으로 용출)에 의하여 정제시켜 갈색 발포체로서 표제생성물 0.322g (57.5%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1- in dimethylformamide (10 mL) Azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (500 mg, 1.03 mmol, cooled (5 ° C.) solution was prepared with 4-[(chloromethyl) thio] benzamide (0.239 g, 1.19 m). Mol), LiI (0.414 g, 3.09 mmol) and N, N-diisopropylethylamine (0.233 g, 1.8 mmol) were added dropwise The mixture was stirred at 5 ° C. for 2 hours and at 20 ° C. for 16 hours. Dilute with ethyl acetate, stir vigorously with cooled dilute HCl and filter, separate the two solution phases and extract the aqueous solution as ethyl acetate The combined organic extracts are washed with brine and dried ( MgSO ₄ ) Evaporated The crude product was purified by silica gel chromatography (eluted first with ethyl acetate and then with CH ₃ CN) to give 0.322 g (57.5%) of the title product as a brown foam.

[단계 C][Step C]

소듐(4R,5S,6S)-3-[[[(p-카르바모일페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Ix)Sodium (4R, 5S, 6S) -3-[[[(p-carbamoylphenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo -1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Ix)

테트라히드로푸란 (40㎖)내 p-니트로페닐 (4R,5S,6S)-3-[[[(p-카르바모일페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.543g, 1m㏖) 용액을 Et₂O (40㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액 (pH7.0, 20㎖)의 혼합물에 첨가하였다. 혼합물을 10% Pd/C 촉매 (0.543g) 위에서 40psi H₂에 3시간동안 가수소분해반응시켰다. 그후 촉매를 여과시킨다음 여과물을 Et₂O로서 추출하였다. 수성상을 0-10% CH₃CN/물로서 용출된 역상 실리카겔에서 크로마토그래피하였다. 관련분율을 결합하고 친액화하여 표제화합물 0.130g (30.2%)을 제공하였다.P-nitrophenyl (4R, 5S, 6S) -3-[[[(p-carbamoylphenyl) thio] methyl] thio] -6- [1 '(R) -hydride in tetrahydrofuran (40 mL) Oxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.543 g, 1 mmol) was dissolved in Et ₂ O (40 mL) and 0.1 To a mixture of M NaH ₂ PO ₄ / NaOH buffer (pH 7.0, 20 mL) was added. The mixture was hydrolyzed at 40 psi H ₂ for 3 hours on 10% Pd / C catalyst (0.543 g). The catalyst was then filtered off and the filtrate was extracted as Et ₂ O. The aqueous phase was chromatographed on reversed phase silica gel eluted as 0-10% CH ₃ CN / water. Relevant fractions were combined and lyophilised to give 0.130 g (30.2%) of the title compound.

[실시예 27]Example 27

소듐 (4R,5S,6S)-3-[[[[(p-아미노메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Iy)Sodium (4R, 5S, 6S) -3-[[[[(p-aminomethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7- Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iy)

[단계 A][Step A]

4-(메틸티오)벤질아지드4- (methylthio) benzylazide

CH₂Cl₂(60㎖)내 4-메틸티오벤질 알코올 (3.0g, 19.45m㏖), 냉각(5℃) 용액을 트리에틸아민 (2.17g, 21.4m㏖) 및 메탄술폰일 클로라이드 (2.45g, 21.4m㏖)로서 처리하였다. 혼합물을 1시간동안 교반한 다음, Et₂O로서 희석하고 여과시켰다. 여과물을 증발건조시켰고, CH₃CN에 재용해시킨다음 NaN₃(1.52g, 23.34m㏖) 및 (Bu)₄NCl(0.150g)으로 처리하였다. 혼합물을 상온에서 18시간 교반한다음, 초산에틸로 희석하였고 물, 묽은 NaHCO₃및 염수로 연속세척하였고, 건조시킨다음 (MgSO₄), 끝으로 증발시켰다. 원 생성물을 실리카겔 크로마토그래피 (석유 에테르 및 CH₂Cl로 용출)에 의하여 정제시켜 운동성 오일로서 표제생성물 2.79g (80.0%)을 제공하였다.4-methylthiobenzyl alcohol (3.0 g, 19.45 mmol), cooled (5 ° C.) solution in CH ₂ Cl ₂ (60 mL) was diluted with triethylamine (2.17 g, 21.4 mmol) and methanesulfonyl chloride (2.45 g , 21.4 mmol). The mixture was stirred for 1 h, then diluted with Et ₂ O and filtered. The filtrate was evaporated to dryness, redissolved in CH ₃ CN and treated with NaN ₃ (1.52 g, 23.34 mmol) and (Bu) ₄ NCl (0.150 g). The mixture was stirred at room temperature for 18 hours, diluted with ethyl acetate, washed successively with water, diluted NaHCO ₃ and brine, dried (MgSO ₄ ) and finally evaporated. The crude product was purified by silica gel chromatography (eluted with petroleum ether and CH ₂ Cl) to give 2.79 g (80.0%) of the title product as a kinetic oil.

[단계 B][Step B]

4-[(클로로메틸)티오]벤질아지드4-[(chloromethyl) thio] benzylazide

CH₂Cl₂내 4-(메틸티오)벤질아지드 (0.359g, 2m㏖) 용액을 얼음에서 냉각시켰고 SO₂Cl₂(0.297g, 2.2m㏖)로서 천천히 처리하였다. 반응 혼합물을 5℃에서 1.5 시간동안 교반한다음 증발건조시켰다. 원화합물을 황색오일로서 얻었다.A solution of 4- (methylthio) benzylazide (0.359 g, 2 mmol) in CH ₂ Cl ₂ was cooled on ice and treated slowly with SO ₂ Cl ₂ (0.297 g, 2.2 mmol). The reaction mixture was stirred at 5 ° C. for 1.5 h and then evaporated to dryness. The original compound was obtained as a yellow oil.

[단계 C][Step C]

p-니트로벤질 (4R,5S,6S)-3-[[[[(p-아지도메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[[(p-azidomethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4- Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF (15㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (849㎎, 1.75m㏖) 냉각 (5℃) 용액을 4-[(클로로메틸)티오]벤질아지드 (0.417g, 2m㏖), LiI (0.703g, 5.25m㏖) 및 N,N-디이소프로필에틸아민 (0.396g, 3m㏖)으로 처리하였다. 5℃에서 20시간 교반후, 용액을 초산에틸로 희석하였고 냉각된 묽은 HCl 수용액으로 세게 교반하였다. 혼합물을 여과시켰고 유기상을 분리하였다. 수성상을 초산에틸로 추출하였다. 유기상을 결합하고, 염수로 세척하고, 건조시킨다음 (MgSO₄) 증발시켰다. 원생성물을 실리카겔 크로마토그래피(0-10% CH₃CN/CH₂Cl₂로서 용출)에 의하여 정제시켜 표제생성물 0.485g (49.9%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1-azabicyclo in DMF (15 mL) [3.2.0] heptan-2-ene-2-carboxylate (849 mg, 1.75 mmol) Cooled (5 ° C.) solution was purified with 4-[(chloromethyl) thio] benzylazide (0.417 g, 2 mmol). , LiI (0.703 g, 5.25 mmol) and N, N-diisopropylethylamine (0.396 g, 3 mmol). After stirring at 5 ° C. for 20 hours, the solution was diluted with ethyl acetate and vigorously stirred with cold diluted aqueous HCl solution. The mixture was filtered and the organic phase was separated. The aqueous phase was extracted with ethyl acetate. The organic phases are combined, washed with brine, dried (MgSO ₄ ) and evaporated. The crude product was purified by silica gel chromatography (eluted as 0-10% CH ₃ CN / CH ₂ Cl ₂ ) to give 0.485 g (49.9%) of the title product.

[단계 D][Step D]

소듐(4R,5S,6S)-3-[[[[(p-아미도메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Iy)Sodium (4R, 5S, 6S) -3-[[[[(p-amidomethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iy)

테트라히드로푸란내 p-니트로벤질 (4R,5S,6S)-3-[[[[(p-아지도메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.470g, 0.846m㏖) 용액을 Et₂O (32㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액 (pH7.0, 16㎖)의 혼합물에 첨가하였다. 혼합물을 10% Pd/C 촉매위에서 40psi H₂에 3.75 시간동안 가수소분해반응시켰다. 그후 촉매를 여과시켰고 여과물을 Et₂O로 추출하였다. 수성상을 10-25% CH₃CN/물로 용출된 역상 실리카 겔에서 크로마토그래피하였다. 관련분율을 결합하고 친액화하여 표제화합물 0.065g (19.5%) 을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -3-[[[[(p-azidomethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl in tetrahydrofuran ] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.470 g, 0.846 mmol) was dissolved in Et ₂ O (32 mL) and 0.1 M. To a mixture of NaH ₂ PO ₄ / NaOH buffer (pH 7.0, 16 mL) was added. The mixture was hydrolyzed at 40 psi H ₂ for 3.75 hours on a 10% Pd / C catalyst. The catalyst was then filtered and the filtrate was extracted with Et ₂ O. The aqueous phase was chromatographed on reversed phase silica gel eluted with 10-25% CH ₃ CN / water. Relevant fractions were combined and lyophilised to give 0.065 g (19.5%) of the title compound.

[실시예 28]Example 28

소듐(4R,5S,6S)-3-[[[[(p-히드록시메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Iz)Sodium (4R, 5S, 6S) -3-[[[[((p-hydroxymethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7 -Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iz)

p-(메틸티오)벤질 p-니트로벤질 카르보네이트p- (methylthio) benzyl p-nitrobenzyl carbonate

CH₃CN (25㎖)내 p-(메틸티오)벤질 알코올 (1.0g, 6.48m㏖), 카르보닐 디이미다졸 (1.16g, 7.13m㏖) 및 나트륨 이미다졸 (0.010g) 용액을 20℃ 에서 30분간 교반하였고, 이어서 p-니트로벤질 알코올 (1.09g, 7.13m㏖)을 첨가하였다. 교반을 추가로 24시간 계속하였고 반응 혼합물을 증발건조시켰다. 원 카르보네이트를 실리카겔 크로마토그래피 (CH₂Cl₂)로 용출)에 의하여 정제시켜 백색고체로서 표제화합물 2.13g (98.6%)을 제공하였다. 융점 84-86℃.A solution of p- (methylthio) benzyl alcohol (1.0 g, 6.48 mmol), carbonyl diimidazole (1.16 g, 7.13 mmol) and sodium imidazole (0.010 g) in CH ₃ CN (25 mL) was heated to 20 ° C. Stirred for 30 min, then p-nitrobenzyl alcohol (1.09 g, 7.13 mmol) was added. Stirring was continued for another 24 hours and the reaction mixture was evaporated to dryness. The original carbonate was purified by silica gel chromatography (eluted with CH ₂ Cl ₂ ) to give 2.13 g (98.6%) of the title compound as a white solid. Melting point 84-86 ° C.

[단계 B][Step B]

p-[(클로로메틸)티오]벤질 p-니트로벤질 카르보네이트p-[(chloromethyl) thio] benzyl p-nitrobenzyl carbonate

CH₂Cl₂(50㎖)내 p-(메틸티오)벤질 p-니트로벤질 카르보네이트 (1.0g, 3m㏖) 냉각 (5℃) 용액을 SO₂Cl₂(0.425g, 3.15m㏖)로서 적가 처리하였다. 5℃에서 얻어진 용액을 30분간 교반한후, 용매를 증발시켜 고체화된 오일로서 표제화합물 (1.10g, 수율. 99.7%)을 남겼다. 그 고체를 다음단계에 정제없이 사용하였다.P- (methylthio) benzyl p-nitrobenzyl carbonate (1.0 g, 3 mmol) cooled (5 ° C.) solution in CH ₂ Cl ₂ (50 mL) was treated as SO ₂ Cl ₂ (0.425 g, 3.15 mmol). It was added dropwise. After stirring the solution obtained at 5 ° C. for 30 min, the solvent was evaporated to leave the title compound (1.10 g, yield. 99.7%) as a solidified oil. The solid was used without purification in the next step.

[단계 C][Step C]

p-니트로벤질 (4R,5S,6S)-3-[[[[4-[[[[(p-니트로벤질)옥시]카르보닐]옥시]메틸]페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[[4-[[[[(p-nitrobenzyl) oxy] carbonyl] oxy] methyl] phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

디메틸포름아미드 (15㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.849g, 1.75m㏖) 냉각 (5℃) 용액을 p-[(클로로메틸)티오]벤질 p-니트로벤질카르보네이트 (0.736g, 2m㏖), LiI (0.703g, 5.25 m㏖) 및 N,N-디이소프로필에틸아민 (0.396g, 3m㏖)으로서 처리하였다. 반응 혼합물을 5℃ 에서 18시간 교반한다음, 초산에틸로 희석하였고 냉각된 묽은 HCl과 함께 세게 흔들었다. 혼합물을 여과시켰고, 유기상을 분리한다음 수성층을 초산에틸로서 추출하였다. 유기 추출물을 결합하고, 염수로 세척하며, 건조시킨다음 (MgSO₄) 증발시켰다. 원 생성물을 실리카겔 크로마토그래피 (0-10% CH₃CN/CH₂Cl₂로서 용출)에 의하여 정제시켰다. 관련분율을 결합하였고 증발시켜 발포체로서 표제화합물 0.638g (51.4%)을 남겼다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1- in dimethylformamide (15 mL) Azabicyclo [3.2.0] heptane-2-ene-2-carboxylate (0.849 g, 1.75 mmol) cooled (5 ° C.) solution to p-[(chloromethyl) thio] benzyl p-nitrobenzylcarbonate (0.736 g, 2 mmol), LiI (0.703 g, 5.25 mmol) and N, N-diisopropylethylamine (0.396 g, 3 mmol) were treated. The reaction mixture was stirred for 18 h at 5 ° C., then diluted with ethyl acetate and vigorously shaken with cold dilute HCl. The mixture was filtered, the organic phase was separated and the aqueous layer was extracted as ethyl acetate. The organic extracts are combined, washed with brine, dried (MgSO ₄ ) and evaporated. The crude product was purified by silica gel chromatography (eluted as 0-10% CH ₃ CN / CH ₂ Cl ₂ ). Relevant fractions were combined and evaporated to leave 0.638 g (51.4%) of the title compound as a foam.

[단계 D][Step D]

소듐 (4R,5S,6S)-3-[[[[(p-히드록시메틸)페닐]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Iz)Sodium (4R, 5S, 6S) -3-[[[[(p-hydroxymethyl) phenyl] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo -1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iz)

테트라히드로푸란 (35㎖)내 p-니트로벤질 (4R,5S,6S)-3-[[[[4-[[[[(p-니트로벤질)옥시]카르보닐]옥시]메틸]페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.635g, 0.895m㏖) 용액을 Et₂O (35㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액 (17.8㎖, pH 7.0)의 혼합물에 첨가하였다. 혼합물을 10% Pd/C 촉매 (0.60g) 위에서 40psi H₂에 3 시간동안 가수소분해반응시켰다. 그후 촉매를 여과시켰고 여과물을 Et₂O로 추출하였다. 수성상을 0-10% CH₃CN/물로서 용출된 역상 실리카겔 (partisil)에서 크로마토그래피하였다. 관련분율을 친액화하여 백색 발포체로서 표제 생성물 0.08g (21.4%)을 얻었다.P-nitrobenzyl (4R, 5S, 6S) -3-[[[[4-[[[(p-nitrobenzyl) oxy] carbonyl] oxy] methyl] phenyl] thio in tetrahydrofuran (35 mL) ] Methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.635 g, 0.895 mmol) was added to a mixture of Et ₂ O (35 mL) and 0.1 M NaH ₂ PO ₄ / NaOH buffer (17.8 mL, pH 7.0). The mixture was hydrolyzed at 40 psi H ₂ for 3 hours on a 10% Pd / C catalyst (0.60 g). The catalyst was then filtered and the filtrate was extracted with Et ₂ O. The aqueous phase was chromatographed on reversed phase silica gel eluted as 0-10% CH ₃ CN / water. Relevant fractions were lysed to yield 0.08 g (21.4%) of the title product as a white foam.

[실시예 29]Example 29

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(1-메틸테트라졸-5-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Iaa)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(1-methyltetrazol-5-yl) thio] methyl] thio] 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Iaa)

[단계 A][Step A]

5-[(클로로메틸)티오]-1-메틸-테트라졸5-[(chloromethyl) thio] -1-methyl-tetrazole

CH₃CN (50㎖)내 나트륨 5-메르캅토-1-메틸테트라졸히드레이트 (1.38g, 10.0m㏖) 용액을 얼음에서 냉각시켰고 브로모클로로메탄 (6.47g, 50m㏖) 일부분으로 처리하였다. 얼음조를 제거하였고 반응 혼합물을 상온에서 18시간 교반하였다. 그후 용매를 증발시켰고 원생성물을 실리카겔 (CH₂Cl₂로서 용출)위 크로마토그래피에 의하여 정제시켜 표제화합물 1.27g (77%)을 제공하였다 : m.p. 55-57℃.A solution of sodium 5-mercapto-1-methyltetrazolhydrate (1.38 g, 10.0 mmol) in CH ₃ CN (50 mL) was cooled in ice and treated with a portion of bromochloromethane (6.47 g, 50 mmol). . The ice bath was removed and the reaction mixture was stirred for 18 hours at room temperature. The solvent was then evaporated and the crude product was purified by chromatography on silica gel (eluted as CH ₂ Cl ₂ ) to give 1.27 g (77%) of the title compound: mp 55-57 ° C.

[단계 B][Step B]

p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(1-메틸테트라졸-5-일)티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(1-methyltetrazol-5-yl) thio] -7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

DMF (20㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (970㎎, 2m㏖) 냉각용액 (5℃)을 5-[(클로로메틸)티오]-1-메틸-테트라졸 (362㎎, 2.2m㏖), 요오드화 리튬 (803㎎, 6m㏖) 및 DIPEA (427㎎, 3.3m㏖)로서 처리하였다. 반응혼합물을 5℃ 에서 20시간 교반한다음, EtOAc로서 희석하였고, 냉각된 묽은 HCl과 함께 세게 교반하였고 여과하였다. 유기상을 분리하였고 염수로서 3회 세척하였다. 건조 (MgSO₄) 후, 용매를 증발시킨다음 원 디티오아세탈을 실리카겔 [CH₃CN (O → 15%)/CH₂Cl₂로 용출]위 크로마토그래피에 의하여 정제시켜 표제생성물 275㎎ (27.1%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1-azabicyclo in DMF (20 mL) [3.2.0] Heptane-2-ene-2-carboxylate (970 mg, 2 mmol) cooling solution (5 ° C.) was added 5-[(chloromethyl) thio] -1-methyl-tetrazole (362 mg, 2.2 mmol), lithium iodide (803 mg, 6 mmol) and DIPEA (427 mg, 3.3 mmol). The reaction mixture was stirred for 20 h at 5 ° C., then diluted as EtOAc, stirred vigorously with cooled dilute HCl and filtered. The organic phase was separated and washed three times with brine. After drying (MgSO ₄ ), the solvent is evaporated and the original dithioacetal is purified by chromatography over silica gel [eluted with CH ₃ CN (O → 15%) / CH ₂ Cl ₂ ] to give the title product 275 mg (27.1%). ).

[단계 C][Step C]

THF (20㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(1-메틸테트라졸-5-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (270㎎, 0.53m㏖) 용액을 Et₂O (20㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액 (pH 7.0, 10㎖) 의 혼합물에 첨가하였다. 이 혼합물을 10% Pd/C (270㎎) 촉매위에서 40psi H₂에 4시간 가수소분해반응 시켰다. 그후 촉매를 여과시켰고 여과물을 Et₂O 로서 추출하였다. 수성산을 처음에 H₂O 및 그후 H₂O 내 5% CH₃CN으로 용출된 역상 실리카겔에서 크로마토그래피하였다. 관련분율을 결합하고 친액화하여 표제생성물 52㎎ (24.8%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(1-methyltetrazol-5-) in THF (20 mL) Yl] thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (270 mg, 0.53 mmol) was dissolved in Et ₂ O (20 mL). And 0.1M NaH ₂ PO ₄ / NaOH buffer (pH 7.0, 10 mL). The mixture was hydrolyzed at 40 psi H ₂ for 4 hours on 10% Pd / C (270 mg) catalyst. The catalyst was then filtered and the filtrate was extracted as Et ₂ O. The aqueous acid was first chromatographed on reversed phase silica gel eluted with H ₂ O and then with 5% CH ₃ CN in H ₂ O. Relevant fractions were combined and lyophilised to give 52 mg (24.8%) of the title product.

[실시예 30]Example 30

소듐 (4R,5S,6S)-3-[[[[(p-히드라지노카르보닐)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Ibb)Sodium (4R, 5S, 6S) -3-[[[[(p-hydrazinocarbonyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Ibb)

[단계 A][Step A]

4-[(클로로메틸)티오]벤조산 히드라지드4-[(chloromethyl) thio] benzoic acid hydrazide

CH₃CN (15㎖)내 히드라진 (0.64g, 20m㏖) 용액에 상온에서 CH₃CN (10㎖)내 4-[(클로로메틸)티오]벤조일클로라이드 (2.0g, 9.11m㏖) 용액을 적가하였다. 백색고체의 형성을 초래하면서 발열반응이 일어났다. 혼합물을 30분간 교반한다음 증발건조시켰다. 원생성물을 실리카겔 크로마토그래피 (EtOAc로서 용출)에 의하여 정제시켜 백색고체로서 표제의 히드라지드 0.758g (38.4%)을 제공하였다.CH ₃ CN (15㎖) in hydrazine _{(0.64g, 20m㏖) CH 3 CN} (10㎖) was added at room temperature within 4 - [(chloromethyl) thio] benzoyl chloride (2.0g, 9.11m㏖) was added dropwise a solution It was. An exothermic reaction occurred, leading to the formation of a white solid. The mixture was stirred for 30 minutes and then evaporated to dryness. The crude product was purified by silica gel chromatography (eluted as EtOAc) to give 0.758 g (38.4%) of the title hydrazide as a white solid.

[단계 B][Step B]

p-니트로벤질 2-[p-[(클로로메틸)티오]벤조일]히드라진 카르복실레이트p-nitrobenzyl 2- [p-[(chloromethyl) thio] benzoyl] hydrazine carboxylate

CH₃CN (25㎖)내 4-[(클로로메틸)티오]벤조산 히드라지드 (0.433g, 2m㏖) 및 p-니트로벤질클로로포르메이트 (0.475g, 2.2m㏖) 냉각 (5℃) 현탁액에 N,N-디이소프로필에틸아민 (0.285g, 2.2m㏖) 을 적가하였다. 반응혼합물을 5℃에서 15분간 및 20℃에서 다시 15분간 교반하였다. 그후 용매를 증발시켰고 얻어진 원생성물을 실리카겔 크로마토그래피 (0-10% CH₃CN/CH₂Cl₂로서 용출)에 의하여 정제하였다. 관련분율을 결합한다음 증발시켜 황색고체로서 표제생성물 0.34g (42.9%)을 제공하였다.To a suspension of 4-[(chloromethyl) thio] benzoic acid hydrazide (0.433 g, 2 mmol) and p-nitrobenzylchloroformate (0.475 g, 2.2 mmol) in CH ₃ CN (25 mL) cooled (5 ° C.) N, N-diisopropylethylamine (0.285 g, 2.2 mmol) was added dropwise. The reaction mixture was stirred at 5 ° C. for 15 minutes and again at 20 ° C. for 15 minutes. The solvent was then evaporated and the resulting crude product was purified by silica gel chromatography (eluted as 0-10% CH ₃ CN / CH ₂ Cl ₂ ). The relevant fractions were combined and evaporated to give 0.34 g (42.9%) of the title product as a yellow solid.

[단계 C][Step C]

p-니트로벤질 (4R,5S,6S)-3-[[[4-[2-[[(p-니트로벤질)옥시]카르보닐]히드라지노]페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트p-nitrobenzyl (4R, 5S, 6S) -3-[[[4- [2-[[(p-nitrobenzyl) oxy] carbonyl] hydrazino] phenyl] thio] methyl] thio] -6- [ 1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

디메틸포름아미드 (10㎖)내 p-니트로벤질 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-은 메르캅토-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.379g, 0.78m㏖) 냉각 (5℃) 용액을 p-니트로벤질 2-[p-[(클로로메틸)티오]벤조일]히드라진-카르복실레이트 (0.34g, 0.86m㏖), LiI (0.345g, 2.58m㏖) 및 N,N-디이소프로필에틸아민 (0.166g, 1.29m㏖)으로 처리하였다. 5℃에서 18시간 교반한후, 용액을 초산에틸로서 희석하였고, 냉각된 묽은 HCl과 함께 세게 흔든다음 여과시켰다. 유기상을 분리하였고 수성층을 초산에틸로서 추출하였다. 추출물을 결합하였고, 염수로서 2회 세척하고, 건조시킨다음 (MgSO₄) 증발건조시켰다. 원 생성물을 실리카겔 크로마토그래피 (0-10% CH₃CN/CH₂Cl₂그후 초산에틸로 용출)에 의하여 정제시켜 황색 발포체로서 표제의 생성물 0.227g (39.5%)을 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3- is mercapto-7-oxo-1- in dimethylformamide (10 mL) Azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.379 g, 0.78 mmol) cooled (5 ° C.) solution to p-nitrobenzyl 2- [p-[(chloromethyl) thio] benzoyl ] Treated with hydrazine-carboxylate (0.34 g, 0.86 mmol), LiI (0.345 g, 2.58 mmol) and N, N-diisopropylethylamine (0.166 g, 1.29 mmol). After stirring for 18 h at 5 ° C., the solution was diluted with ethyl acetate, vigorously shaken with cooled dilute HCl and filtered. The organic phase was separated and the aqueous layer was extracted as ethyl acetate. The extracts were combined, washed twice with brine, dried (MgSO ₄ ) and evaporated to dryness. The crude product was purified by silica gel chromatography (0-10% CH ₃ CN / CH ₂ Cl ₂ then eluted with ethyl acetate) to give 0.227 g (39.5%) of the title product as a yellow foam.

[단계 D][Step D]

테트라히드로푸란 (20㎖)내 p-니트로벤질 (4R,5S,6S)-3-[[[4-[2-[[(p-니트로벤질)옥시]카르보닐]-히드라지노]페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (0.29g, 0.393m㏖) 용액을 Et₂O (20㎖) 및 0.1M NaH₂PO₄/NaOH 완충용액 (10㎖, pH 7.0)의 혼합물에 첨가하였다. 혼합물을 10% Pd/C 촉매 (0.29g) 위에서 40psi H₂에 4시간 가수소분해반응시켰다. 이때, 촉매를 여과시켰고 새배치 (0.175g)로 대체하였다. 가수소분해반응조건을 추가로 2시간 유지하였다. 그후 반응혼합물을 여과시켰고 여과물을 Et₂O로 추출하였다. 수성상을 0-5% CH₃CN/물로 용출된 역상 실리카겔 (파르티실)에서 크로마토그래피하였다. 관련분율을 결합하고 친액화하여 백색 발포체 (0.048g, 27.4%)를 제공하였다.P-nitrobenzyl (4R, 5S, 6S) -3-[[[4- [2-[[(p-nitrobenzyl) oxy] carbonyl] -hydrazino] phenyl] thio in tetrahydrofuran (20 mL) ] Methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (0.29 g, 0.393 mmol) solution was added to a mixture of Et ₂ O (20 mL) and 0.1 M NaH ₂ PO ₄ / NaOH buffer (10 mL, pH 7.0). The mixture was hydrolyzed at 40 psi H ₂ for 4 hours on a 10% Pd / C catalyst (0.29 g). At this time, the catalyst was filtered and replaced with fresh batch (0.175 g). Hydrolysis reaction conditions were maintained for an additional 2 hours. The reaction mixture was then filtered and the filtrate was extracted with Et ₂ O. The aqueous phase was chromatographed on reversed phase silica gel (particyl) eluted with 0-5% CH ₃ CN / water. The relevant fractions were combined and lyophilized to give a white foam (0.048 g, 27.4%).

[실시예 31]Example 31

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-옥사디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Icc)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-oxadiazol-2-yl ) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Icc)

[단계 A][Step A]

2-[(요오드메틸)티오]-5-메틸-1,3,4-옥사디아졸2-[(iodinemethyl) thio] -5-methyl-1,3,4-oxadiazole

CH₃CN (330㎖)내 2-메르캅토-5-메틸-1,3,4-옥사디아졸 칼륨염 (33.0g, 214m㏖) 현탁액을 BrCH₂Cl (130g, 1㏖) 존재하에 상온에서 24시간 교반하였다. 그후 용매를 증발시켰고 상온에서 24시간 교반하였다. 그후 용매를 증발시켰고 잔류물을 H₂O와 EtOAc 사이에 분배시켰다. 유기상을 분리하고, 건조시킨다음 (MgSO₄) 증발시켰다. 이와같이 얻어진 잔류물을 Et₂O 에 재용해시켰고; 에테르 용액을 목탄으로 처리한다음 셀라이트 패드를 통하여 여과시켰다. 여과물의 증발은 무색오일 29.4g을 남겼다. 원유를 아세톤 (350㎖)에 용해시켰고, 요오드화 나트륨 (134g, 894m㏖)을 첨가하였다. 얻어진 혼합물을 24시간 환류하였고, Et₂O로 희석한다음 H₂O로 세척하였다. 수성상을 Et₂O로 재추출하였다. 유기 추출물을 결합하였고, H₂O와 염수로 세척하고, 건조시켰고 (MgSO₄), 목탄으로 처리하고, 여과시킨다음 끝으로 증발시켜 황색오일로서 표제화합물 40.0g (87.3%)을 제공하였다.Suspension of 2-mercapto-5-methyl-1,3,4-oxadiazole potassium salt (33.0 g, 214 mmol) in CH ₃ CN (330 mL) was carried out at room temperature in the presence of BrCH ₂ Cl (130 g, 1 mol). Stir for 24 hours. The solvent was then evaporated and stirred at room temperature for 24 hours. The solvent was then evaporated and the residue was partitioned between H ₂ O and EtOAc. The organic phase is separated, dried (MgSO ₄ ) and evaporated. The residue thus obtained was redissolved in Et ₂ O; The ether solution was treated with charcoal and then filtered through a pad of celite. Evaporation of the filtrate left 29.4 g of colorless oil. Crude oil was dissolved in acetone (350 mL) and sodium iodide (134 g, 894 mmol) was added. The resulting mixture was refluxed for 24 hours, diluted with Et ₂ O and washed with H ₂ O. The aqueous phase was reextracted with Et ₂ O. The organic extracts were combined, washed with H ₂ O and brine, dried (MgSO ₄ ), treated with charcoal, filtered and finally evaporated to afford 40.0 g (87.3%) of the title compound as a yellow oil.

[단계 B][Step B]

알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-옥사디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-oxadiazol-2-yl ) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

THF (1500㎖) 내 알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-(디페닐포스포노)-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (75.9g, 152m㏖) 냉각 (5℃) 용액을 LiSH [10.3g, 258m㏖, LiSH는 헥산 및 THF 동일부피내 BuLi 냉각 (5℃) 용액을 H₂S로 포화시켜 제조되었다. 침전염을 여과시킨다음 건조시켰다.]로서 부분적으로 처리하였다. 용액을 35분간 교반한다음 2-[(요오도메틸)티오]-5-메틸-1,3,4-옥사디아졸 (58.4g, 228m㏖) 이어서 디이소프로필에틸아민 (40㎖, 228m㏖)을 첨가하였다. 반응 혼합물을 30분간 교반하였고, 초산 (14㎖) 및 냉각 H₂O (1000㎖)를 첨가하였다. 생성물을 EtOAc로 추출하였다. 유기추출물을 연속적으로 H₂O, 중탄산나트륨 수용액 및 염수로 세척하였고, 건조시킨다음 (MgSO₄) 증발시켰다. 얻어진 원 디티오아세탈을 CH₂Cl₂내 CH₃CN (0-25% CH₃CN)으로 용출된 실리카겔위 크로마토그래피에 의하여 정제시켜 표제화합물 15.5g (24.8%)을 제공하였다.Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3- (diphenylphosphono) -4-methyl-7-oxo-1-azabicyclo in THF (1500 mL) [3.2.0] Heptane-2-ene-2-carboxylate (75.9 g, 152 mmol) cooled (5 ° C.) solution to LiSH [10.3 g, 258 mmol, LiSH cooled to BuLi in the same volume of hexane and THF (5 C) was prepared by saturating the solution with H ₂ S. The precipitated salts were filtered off and dried. The solution was stirred for 35 minutes and then 2-[(iodomethyl) thio] -5-methyl-1,3,4-oxadiazole (58.4 g, 228 mmol) followed by diisopropylethylamine (40 mL, 228 mmol ) Was added. The reaction mixture was stirred for 30 minutes, acetic acid (14 mL) and cold H ₂ O (1000 mL) were added. The product was extracted with EtOAc. The organic extract was washed successively with H ₂ O, aqueous sodium bicarbonate solution and brine, dried (MgSO ₄ ) and evaporated. The resulting dithioacetal was purified by chromatography on silica gel eluted with CH ₃ CN (0-25% CH ₃ CN) in CH ₂ Cl _{2 to} give 15.5 g (24.8%) of the title compound.

[단계 C][Step C]

CH₂Cl₂(130㎖)내 알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-옥사디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (15.35g, 37.3m㏖) 용액을 5℃로 냉각시켰고 연속적으로 Pd (PPh₃)₄(1.0g, 0.87m㏖), PPh₃(100㎎, 0.38m㏖) 및 EtOAc내 나트륨 에틸 헥산오에이트 0.5M (74.6㎖, 37.3m㏖) 용액을 적가처리하였다. 5℃ 에서 90분간 교반한 후, 반응 혼합물을 냉각 H₂O (2 × 150㎖)로 추출하였다. 수용액을 H₂O 내 CH₃CN (O → 20% CH₃CN)으로 용출된 역상실리카겔 (BondaPak C-18)에서 크로마토그래피하였다. 관련분율의 친액화는 필요로한 생성물 13.0g (88.6%)을 제공하였다.Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,) in CH ₂ Cl ₂ (130 mL) 4-oxadiazol-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (15.35 g, 37.3 mmol) solution Was cooled to 5 ° C. and successively Pd (PPh ₃ ) ₄ (1.0 g, 0.87 mmol), PPh ₃ (100 mg, 0.38 mmol) and 0.5 M (74.6 mL, 37.3 mmol) of sodium ethyl hexaneoate in EtOAc ) Solution was added dropwise. After stirring at 5 ° C. for 90 min, the reaction mixture was extracted with cold H ₂ O (2 × 150 mL). The aqueous solution was chromatographed on reversed phase silica gel (BondaPak C-18) eluted with CH ₃ CN (O → 20% CH ₃ CN) in H ₂ O. Relevance of the relevant fractions gave 13.0 g (88.6%) of the required product.

[실시예 32]Example 32

소듐 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-티아디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (Idd)Sodium (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-thiadiazol-2-yl ) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate (Idd)

[단계 A][Step A]

2-[(요오도메틸)티오]-5-메틸-1,3,4-티아디아졸2-[(iodomethyl) thio] -5-methyl-1,3,4-thiadiazole

2-메르캅토-5-메틸-1,3,4-티아디아졸 (41.4g, 313m㏖)을 EtOH (160㎖)내 85% KOH (20.7g, 313m㏖) 얼음조냉각 (5℃) 수용액에 첨가하였다. 얼음조를 제거하였고 혼합물을 완전한 용해가 일어날때까지 (30분) 교반하였다. 그후 용매를 증발시켜 고체화된 점성오일을 남겼다. 얻어진 칼륨염을 CH₃CN (200㎖)에 용해시켰고, 용액을 얼음조에서 냉각시킨다음 브로모클로로메탄 (121g, 939m㏖)으로 처리하였다. 얼음조에서 얼음을 용융시켰고 추가로 혼합물을 상온에서 25시간 교반하였다. 반응 혼합물을 EtOAc와 H₂O 사이에 분배시켰다. 유기상을 건조시킨다음 (MgSO₄) 증발시켰다. 잔류물을 처음에 CH₂Cl₂및 그후 5% CH₃CN/CH₂Cl₂로 용출된 실리카겔 패드로 통과시켜 정제하여 2-[(클로로메틸)티오]-5-메틸-1,3,4-티아디아졸 93.0g (76.0%)을 제공하였다.2-mercapto-5-methyl-1,3,4-thiadiazole (41.4 g, 313 mmol) in 85% KOH (20.7 g, 313 mmol) ice bath (5 ° C.) solution in EtOH (160 mL) Was added. The ice bath was removed and the mixture was stirred (30 min) until complete dissolution occurred. The solvent was then evaporated to leave a solidified viscous oil. The resulting potassium salt was dissolved in CH ₃ CN (200 mL) and the solution was cooled in an ice bath and then treated with bromochloromethane (121 g, 939 mmol). The ice was melted in an ice bath and the mixture was further stirred at room temperature for 25 hours. The reaction mixture was partitioned between EtOAc and H ₂ O. The organic phase is dried (MgSO ₄ ) and evaporated. The residue was purified by passing through a pad of silica gel _first eluted with CH ₂ Cl ₂ and then with 5% CH ₃ CN / CH ₂ Cl ₂ to afford 2-[(chloromethyl) thio] -5-methyl-1,3,4 Provided 93.0 g (76.0%) of thiadiazole.

티아디아졸을 아세톤 (450㎖)에 용해시켰고, 얼음조로서 5℃로 냉각시킨다음 NaI (178g, 1.19㏖)로 천천히 처리하였다. 얼음조에서 얼음을 용융시켰고 혼합물을 상온에서 75시간 교반하였다. 아세톤을 대부분 증발시켰고 잔류슬러리를 Et₂O와 H₂O 사이에 분배시켰다. 에테르상을 NaHSO₃수용액 및 H₂O로 세척하고 건조시킨다음 (MgSO₄), 증발시켰다. 얻어진 원 생성물을 CH₂Cl₂및 CH₃CN(O → 10%)/CH₂Cl₂로서 용출된 실리카겔에서 크로마토그래피하여 표제화합물 50.0g (77.2%)을 제공하였다.Thiadiazole was dissolved in acetone (450 mL), cooled to 5 ° C. as an ice bath and then slowly treated with NaI (178 g, 1.19 mol). The ice was melted in an ice bath and the mixture was stirred at room temperature for 75 hours. Acetone was mostly evaporated and the residual slurry was partitioned between Et ₂ O and H ₂ O. The ether phase was washed with aqueous NaHSO ₃ solution and H ₂ O, dried (MgSO ₄ ) and evaporated. The obtained crude product was chromatographed on silica gel eluted with CH ₂ Cl ₂ and CH ₃ CN (O → 10%) / CH ₂ Cl ₂ to give 50.0 g (77.2%) of the title compound.

[단계 B][Step B]

알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-티아디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-thiadiazol-2-yl ) Thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate

THF (2ℓ)내 알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-3-(디페닐포스포노)-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 (100g, 0.2㏖) 냉각 (5℃) 용액을 LiSH (12.0g, 0.3㏖)로서 부분처리하였다. 25분간 교반한후, 2-[(요오도메틸)티오]-5-메틸-1,3,4-티아디아졸 (50g, 0.184㏖) 이어서 DIPEA (25.9g, 200m㏖) 을 첨가하였다. 반응 혼합물을 0℃ 에서 30분간 상온에서 30분간 교반하였다. 초산 (15㎖) 이어서 냉각 H₂O (2ℓ)를 반응 혼합물에 첨가하였다. 수용액을 EtOAc로서 추출하였다. 유기추출물을 결합하였고, H₂O, NaHCO₃및 염수로 연속세척하고, 건조시킨다음 (MgSO₄), 증발시켰다. 얻어진 원 생성물을 실리카겔 크로마토그래피 [CH₃CN(O → 60%)/CH₂Cl₂로 용출]에 의하여 정제하였다. 관련 분율을 결합하였고 증발시켜 고체를 남겼고 Et₂O 에서 분쇄하고 여과에 의하여 수집하여 표제화합물 29.93g (28.0%)을 제공하였다.Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -3- (diphenylphosphono) -4-methyl-7-oxo-1-azabicyclo [in THF (2 L) 3.2.0] heptane-2-ene-2-carboxylate (100 g, 0.2 mol) The cooled (5 ° C.) solution was partially treated with LiSH (12.0 g, 0.3 mol). After stirring for 25 minutes, 2-[(iodomethyl) thio] -5-methyl-1,3,4-thiadiazole (50 g, 0.184 mol) was added followed by DIPEA (25.9 g, 200 mmol). The reaction mixture was stirred at 0 ° C. for 30 minutes at room temperature for 30 minutes. Acetic acid (15 mL) then cooled H ₂ O (2 L) was added to the reaction mixture. The aqueous solution was extracted as EtOAc. The organic extracts were combined, washed successively with H ₂ O, NaHCO ₃ and brine, dried (MgSO ₄ ) and evaporated. The obtained crude product was purified by silica gel chromatography [eluted with CH ₃ CN (O → 60%) / CH ₂ Cl ₂ ]. The relevant fractions were combined and evaporated to leave a solid which was triturated in Et ₂ O and collected by filtration to give 29.93 g (28.0%) of the title compound.

[단계 C][Step C]

CH₂Cl₂(200㎖)내 알릴 (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-옥사디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실레이트 냉각 (5℃) 용액을 연속적으로 PPh₃(150㎎, 0.57m㏖), Pd (PPh₃)₄(1.3g, 1.13m㏖) 및 EtOAc내 에틸헥산오에이트 (112㎖, 56m㏖) 0.5M 용액으로 처리하였다. 반응 혼합물을 고체가 침전되는 시간동안 5℃ 에서 2.5시간 교반하였다. 용액을 아세톤 (300㎖)으로 희석한후, 침전물을 여과에 의하여 수집하였고, 세척한다음 (아세톤) 건조시켰다. 이러한 원 생성물 (~30g)을 처음에 H₂O 및 그후 CH₃CN(O → 10%)/H₂O 로서 용출된 역상 실리카겔 (μBondaPak C-18)에서 크로마토그래피에 의하여 정제하였다. 친액화후, 표제화합물 18.9g (82.5%)을 백색고체로서 회수하였다.Allyl (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,) in CH ₂ Cl ₂ (200 mL) 4-oxadiazol-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylate cooling (5 ° C.) solution continuously Treated with PPh ₃ (150 mg, 0.57 mmol), Pd (PPh ₃ ) ₄ (1.3 g, 1.13 mmol) and a 0.5 M solution of ethyl hexaneoate (112 mL, 56 mmol) in EtOAc. The reaction mixture was stirred for 2.5 hours at 5 ° C. during which time the solid precipitated. After diluting the solution with acetone (300 mL), the precipitate was collected by filtration, washed and dried (acetone). This crude product (˜30 g) was purified by chromatography on reverse phase silica gel (μBondaPak C-18) eluted first with H ₂ O and then with CH ₃ CN (O → 10%) / H ₂ O. After lyophilization, 18.9 g (82.5%) of the title compound was recovered as a white solid.

[실시예 33]Example 33

[생물학적 활성][Biological activity]

본 발명의 카르바페넴에 대한 강력한 항균활성 및 중요한 경구 생물이용성을 예시하기 위하여, 표 I, II 및 III은 시험관내 활성 (MIC), 경구 생체내 활성 (PD₅₀) 및 대표약물의 경구투여후 혈액수준 (C_max, T1/2 및 AUC)에 대한 각 데이타를 보여준다.To illustrate the potent antimicrobial activity and important oral bioavailability of carbapenem of the present invention, Tables I, II and III show in vitro activity (MIC), oral in vivo activity (PD ₅₀ ) and after oral administration of representative drugs. Each data is shown for blood levels (C _max , T1 / 2 and AUC).

[I. 시험관내 활성][I. In vitro activity]

표 I은 대표 항균제의 최소 억제농도 (MIC)를 보여준다. 뉴트리언트 (Nutrient)즙 및 박테리아의 밤새 배양체로부터 약 500,000 CFU/㎖ 의 최종 박테리아 접종체를 사용한 마이크로타이터 즙 희석을 이용하여 측정하였다. 그후 마이크로타이터 접시를 35℃ 에서 밤새 배양시켰다. 박테리아의 외관성장을 억제하는 약물의 최저농도로서 ㎍/㎖로 MIC를 측정하였다.Table I shows the minimum inhibitory concentrations (MIC) of representative antimicrobials. Measurements were made using microtiter juice dilution using approximately 500,000 CFU / mL final bacterial inoculum from overnight cultures of Nutrient juice and bacteria. The microtiter dish was then incubated overnight at 35 ° C. MIC was measured at μg / ml as the lowest concentration of drug that inhibited the apparent growth of bacteria.

[II. 경구 6생체내 활성][II. 6 Oral In vivo Activity]

여러가지 박테리아 현탁액 0.5㎖로서 복강내 감염시킨 새앙쥐에 경구투여후 대표 화합물의 생체내 치료효능을 표 II에 제시한다. 그 값을 PD₅₀으로 제공한다 (감염된 새앙쥐의 50%를 보호하는 투여량 ㎎/㎏).The in vivo therapeutic efficacy of representative compounds after oral administration to birds intraperitoneally as 0.5 ml of various bacterial suspensions is shown in Table II. The value is given as PD ₅₀ (dose mg / kg protecting 50% of infected birds).

[III. 약동학][III. Pharmacokinetics]

새앙쥐에서 50㎎/㎏의 경구투여후 본 발명의 선택화합물에 대한 혈액수준 및 반감기를 표 III에 제시한다.Blood levels and half-lives for the selective compounds of the invention following oral administration of 50 mg / kg in rats are shown in Table III.

Claims

다음 일반식의 화합물 또는 비독성의 약리적으로 허용되는 염 또는, 생리학적으로 가수분해 가능한 그의 에스테르.A compound of the following general formula or a nontoxic pharmacologically acceptable salt or physiologically hydrolysable ester thereof.

상기식에서 R¹은 수소 또는 C_1-6알킬이며; n은 0, 1 또는 2이며; R³는 수소 또는 C_1-6알킬이며; R²는 C_1-6알킬, 시아노, -CONH₂, -CH₂OH, -CH₂NH₂, -CONHNH₂또는 할로겐 원자수 5개까지, C_1-6알킬 또는 C_1-6알킬록시기로서 임의치환된 페닐메틸, 또는 다음식으로 표시된 라디칼이며In which R ¹ is hydrogen or C _1-6 alkyl; n is 0, 1 or 2; R ³ is hydrogen or C _1-6 alkyl; R ² is C _1-6 alkyl, cyano, -CONH ₂ , -CH ₂ OH, -CH ₂ NH ₂ , -CONHNH ₂ or up to 5 halogen atoms, C _1-6 alkyl or C _1-6 alkyllock Optionally substituted phenylmethyl, or a radical represented by the formula:

제1항에 있어서, R¹이 수소 또는 메틸이며; n이 0 또는 1이며; R³가 수소, 또는 C_1-6알킬이며; R²가 C_1-6알킬, 시아노, -CONH₂, -CH₂OH, -CH₂NH₂, -CONHNH₂또는 할로겐 원자수 5개까지 또는 C_1-6알킬기로서 임의치환된 페닐, 페닐환위에 할로겐 원자수 5개까지 또는 C_1-6알킬기로서 임의치환된 페닐메틸, 또는 다음식으로 표시된 라디칼인 화합물.The compound of claim 1, wherein R ¹ is hydrogen or methyl; n is 0 or 1; R ³ is hydrogen or C _1-6 alkyl; R ² is phenyl, phenyl optionally substituted with C _1-6 alkyl, cyano, -CONH ₂ , -CH ₂ OH, -CH ₂ NH ₂ , -CONHNH ₂ or up to 5 halogen atoms or as a C _1-6 alkyl group A compound which is phenylmethyl optionally substituted with up to 5 halogen atoms or C _1-6 alkyl group at a ring, or a radical represented by the following formula.

상기식에서 p는 0 또는 1이며; X는 피리딘일, 푸릴 또는 다음식의 라디칼이다:In which p is 0 or 1; X is pyridinyl, furyl or a radical of the formula:

(상기식에서 Y는 황 또는 산소이다.)(Wherein Y is sulfur or oxygen)

제2항에 있어서, R¹이 메틸이고, R³가 수소이며 n이 0인 화합물.The compound of claim 2, wherein R ¹ is methyl, R ³ is hydrogen and n is 0. 4.

제3항에 있어서, p가 0이고 R²가 다음식의 라디칼인 화합물.The compound of claim 3, wherein p is 0 and R ² is a radical of the formula: 5.

(Y는 상기에 정의된 것과 같다)(Y is as defined above)

제4항에 있어서, (4R,5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-옥사디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound of claim 4, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-oxadia) Zol-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제4항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(5-메틸-1,3,4-티아디아졸-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound of claim 4, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(5-methyl-1,3,4-thiadia) Zol-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[(p-카르바모일페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -3-[[[(p-carbamoylphenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4- Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[(p-시아노페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -3-[[[(p-cyanophenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[(p-아미노메틸페닐)티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound according to claim 3, wherein (4R, 5S, 6S) -3-[[[(p-aminomethylphenyl) thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl- 7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-[1'(R)-히드록시에틸]-3-[[[(페닐메틸)티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound according to claim 3, wherein (4R, 5S, 6S)-[1 '(R) -hydroxyethyl] -3-[[[(phenylmethyl) thio] methyl] thio] -4-methyl-7-oxo- A compound that is 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[[(p-히드록시메틸)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound according to claim 3, wherein (4R, 5S, 6S) -3-[[[[(p-hydroxymethyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl]- 4-Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(1-메틸테트라졸-5-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(1-methyltetrazol-5-yl) thio] Methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[[(p-히드라지노카르보닐)페닐]티오]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -3-[[[[(p-hydrazinocarbonyl) phenyl] thio] methyl] thio] -6- [1 '(R) -hydroxyethyl] 4-Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methylthio) methyl] thio] -7-oxo-1 Azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3-yl) methyl] thio] methyl ] Thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-3-[[[(3,4-디클로로페닐)티오]메틸]티오]-6-[1'(R)히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -3-[[[(3,4-dichlorophenyl) thio] methyl] thio] -6- [1 '(R) hydroxyethyl] -4-methyl -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(2,3,4,5,6-펜타플루오로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. A compound according to claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[((2,3,4,5,6-pentafluoro) Rophenyl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptane-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-[1'(R)-히드록시에틸]-3-[[(이소프로필티오)메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S)-[1 '(R) -hydroxyethyl] -3-[[(isopropylthio) methyl] thio] -4-methyl-7-oxo-1- A compound that is azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-[1'(R)-히드록시에틸]-3-[[[[(푸란-2-일)메틸]티오]메틸]티오]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S)-[1 '(R) -hydroxyethyl] -3-[[[[(furan-2-yl) methyl] thio] methyl] thio] -4- Methyl-7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리딘-4-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리딘-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-2-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[(피리딘-3-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-[1'(R)-히드록시에틸]-4-메틸-3-[[[(p-클로로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S)-[1 '(R) -hydroxyethyl] -4-methyl-3-[[[(p-chlorophenyl) thio] methyl] thio] -7- A compound which is oxo-1-azabicyclo [3.2.0] heptan-2-ene-carboxylic acid.

제3항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(페닐티오)메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.4. The compound of claim 3, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(phenylthio) methyl] thio] -7-oxo-1 Azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제2항에 있어서, R¹이 메틸이고, R³가 수소이며 n이 1인 화합물.The compound of claim 2, wherein R ¹ is methyl, R ³ is hydrogen and n is 1.

제25항에 있어서, (4R, 5S,6S)-3-[[[(p-클로로페닐)술핀일]메틸]티오]-6-[1'(R)-히드록시에틸]-4-메틸-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The compound of claim 25, wherein (4R, 5S, 6S) -3-[[[(p-chlorophenyl) sulfinyl] methyl] thio] -6- [1 '(R) -hydroxyethyl] -4-methyl -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제25항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[[[(피리딘-3-일)메틸]술핀일]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The method of claim 25, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[[[(pyridin-3-yl) methyl] sulfinyl] Methyl] thio] -7-oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제25항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[(메틸-술핀일)메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.27. The compound of claim 25, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[(methyl-sulfinyl) methyl] thio] -7-oxo -1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제2항에 있어서, R¹과 R³가 수소이고 n이 0인 화합물.The compound of claim 2, wherein R ¹ and R ³ are hydrogen and n is zero.

제29항에 있어서, (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리디-3-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.30. The compound of claim 29, wherein (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[[(pyridin-3-yl) thio] methyl] thio] -7-oxo- A compound that is 1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제29항에 있어서, (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[(메틸티오)메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.30. The compound of claim 29, wherein (5R, 6S) -6- [1 '(R) -hydroxyethyl] -3-[[(methylthio) methyl] thio] -7-oxo-1-azabicyclo [3.2. 0] heptane-2-ene-2-carboxylic acid.

제29항에 있어서, (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리딘-2-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The method of claim 29, wherein (5R, 6S) -6- [1 ′ (R) -hydroxyethyl] -3-[[[(pyridin-2-yl) thio] methyl] thio] -7-oxo-1 Azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제29항에 있어서, (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(피리딘-4-일)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The method of claim 29, wherein (5R, 6S) -6- [1 ′ (R) -hydroxyethyl] -3-[[[(pyridin-4-yl) thio] methyl] thio] -7-oxo-1 Azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제29항에 있어서, (5R,6S)-6-[1'(R)-히드록시에틸]-3-[[[(p-클로로페닐)티오]메틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.The method of claim 29, wherein (5R, 6S) -6- [1 ′ (R) -hydroxyethyl] -3-[[[(p-chlorophenyl) thio] methyl] thio] -7-oxo-1- A compound that is azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제2항에 있어서, R¹이 메틸이고, R³가 메틸이며 n이 0인 화합물.The compound of claim 2, wherein R ¹ is methyl, R ³ is methyl and n is 0. 4.

제35항에 있어서, (4R, 5S,6S)-6-[1'(R-히드록시에틸]-4-메틸-3-[[1(R)-(메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.36. The compound of claim 35, wherein (4R, 5S, 6S) -6- [1 '(R-hydroxyethyl] -4-methyl-3-[[1 (R)-(methylthio) ethyl] thio] -7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제35항에 있어서, (4R, 5S,6S)-6-[1'(R)-히드록시에틸]-4-메틸-3-[[1(S)-메틸티오)에틸]티오]-7-옥소-1-아자비시클로[3.2.0]헵탄-2-엔-2-카르복실산인 화합물.36. The compound of claim 35, wherein (4R, 5S, 6S) -6- [1 '(R) -hydroxyethyl] -4-methyl-3-[[1 (S) -methylthio) ethyl] thio] -7 Oxo-1-azabicyclo [3.2.0] heptan-2-ene-2-carboxylic acid.

제1항 화합물의 향균 유효량과 약리적으로 허용되는 담체 또는 희석제로 이루어진 경구 투여용 항균제 조성물.An antimicrobial composition for oral administration comprising an antimicrobial effective amount of the compound of claim 1 and a pharmacologically acceptable carrier or diluent.