KR101364748B1 - New fatty acids having physiological activity comprising antimicrobial activity, Marine microorganism producing the new fatty acids and producing method of the new fatty acids using the marine microorganism - Google Patents

New fatty acids having physiological activity comprising antimicrobial activity, Marine microorganism producing the new fatty acids and producing method of the new fatty acids using the marine microorganism Download PDF

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KR101364748B1
KR101364748B1 KR1020130149252A KR20130149252A KR101364748B1 KR 101364748 B1 KR101364748 B1 KR 101364748B1 KR 1020130149252 A KR1020130149252 A KR 1020130149252A KR 20130149252 A KR20130149252 A KR 20130149252A KR 101364748 B1 KR101364748 B1 KR 101364748B1
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신희재
이희승
이연주
몬돌
김지혜
이민아
타랙
이종석
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Abstract

본 발명은 항균 활성을 보이며 고지혈증의 치료제로도 사용할 수 있는 하기 화학식 1 ~ 4로 표시되는 지방산 화합물을 제공하며, 이 지방산 화합물을 효율적으로 생산할 수 있는 기탁번호 KCTC 11975BP로 기탁된 해양 미생물 및 염도 12 g/L로 조절된 배지를 이용한 제조방법을 제공한다. 이와 같은 발명에 의하면 다양한 생리활성을 보이는 신규의 지방산 화합물 및 이의 제조방법에 제공되어 항생제 내성 병원균에 의한 위협, 항진균 약물의 제한적 효능 및 최근의 바이오테러리즘의 등장에 맞설 신규 항미생물 제제의 상업적 공급이 가능할 수 있다.
<화학식 1>

Figure 112013110654443-pat00011

상기 화학식 1에서 R1=R2=H 임.
<화학식 2>
Figure 112013110654443-pat00012

상기 화학식 2에서 R=H 임.
<화학식 3>
Figure 112013110654443-pat00013

상기 화학식 3에서 R1=R2=R3=H 임.
<화학식 4>
Figure 112013110654443-pat00014

상기 화학식 4에서 R1=R2=H 임.The present invention provides a fatty acid compound represented by the following Chemical Formulas 1 to 4 showing antimicrobial activity and can also be used as a therapeutic agent for hyperlipidemia, and marine microorganisms and salts deposited with the accession number KCTC 11975BP that can efficiently produce the fatty acid compound 12 It provides a production method using a medium adjusted to g / L. According to the present invention, a novel fatty acid compound exhibiting various physiological activities and a method for preparing the same provide a commercial supply of new antimicrobial agents against the threat of antibiotic resistant pathogens, limited efficacy of antifungal drugs, and the recent emergence of bioterrorism. It may be possible.
&Lt; Formula 1 >
Figure 112013110654443-pat00011

In Formula 1, R 1 = R 2 = H.
(2)
Figure 112013110654443-pat00012

In Formula 2, R = H.
(3)
Figure 112013110654443-pat00013

In Formula 3 R 1 = R 2 = R 3 = H.
&Lt; Formula 4 >
Figure 112013110654443-pat00014

In Formula 4, R 1 = R 2 = H.

Description

항균 활성을 포함한 다양한 생리 활성을 가지는 신규의 지방산, 이 지방산을 생성하는 해양 미생물 및 이 지방산의 제조방법{New fatty acids having physiological activity comprising antimicrobial activity, Marine microorganism producing the new fatty acids and producing method of the new fatty acids using the marine microorganism}New fatty acids having various physiological activities, including antimicrobial activity, marine microorganisms that produce these fatty acids, and methods for preparing these fatty acids.Antimicrobial activity, marine microorganism producing the new fatty acids and producing method of the new fatty acids using the marine microorganism}

본 발명은 신규의 지방산을 생성하는 해양 미생물, 이 해양 미생물로부터 생성된 신규 지방산 및 이 지방산의 제조방법에 관한 것으로서, 구체적으로는 항균 활성 및 고지혈증에 치료효과를 보이는 새로운 지방산을 생성하는 해양 바실러스 (Marine Bacillus)속 미생물 및 이 해양 바실러스 속 미생물에 의해 생성된 신규 지방산 및 이 신규 지방산의 제조 방법에 관한 것이다.The present invention relates to a marine microorganism producing a novel fatty acid, a novel fatty acid produced from the marine microorganism, and a method for preparing the fatty acid. Specifically, the marine bacillus producing a new fatty acid having a therapeutic effect on antimicrobial activity and hyperlipidemia ( Marine Bacillus genus microorganism and the novel fatty acid produced by the marine microorganism of the genus Bacillus, and a method for producing the new fatty acid.

해양생물들은 다양한 화합물을 생성할 뿐만 아니라 여러 가지 생리 활성을 가지고 있어서 미래의 아주 유망한 천연자원으로 인식되고 있다. Marine organisms are recognized as very promising natural resources of the future because they not only produce various compounds but also have various biological activities.

특히 해양 바실러스 속 미생물은 해양 생태계의 어디에나 다양하게 서식하며, 항균물질, 항암제, 생물계면활성제 등을 생성하는 것으로 알려져 있다. In particular, the marine microorganisms of the genus Marine Bacillus are known to produce a variety of antimicrobials, anticancer agents, biosurfactants, etc. inhabit a variety of marine ecosystems.

항생제 내성 병원균에 의한 위협, 항진균 약물의 제한적 효능 및 최근의 바이오테러리즘의 등장에 맞설 신규 항미생물 제제가 긴급히 필요한 실정이다. There is an urgent need for new antimicrobial agents to combat the threat of antibiotic resistant pathogens, the limited efficacy of antifungal drugs and the recent emergence of bioterrorism.

상술한 바와 같은 문제점을 해결하기 위하여 본 발명자들에 의한 지속적인 연구 결과 해양퇴적토, 해양 조류 및 무척추 동물의 표면에서 분리한 해양 미생물들은 항균활성 등의 생리활성을 보이는 이차 대사 산물을 생성하는 것을 확인하였으며, 해양 미생물로부터 생리활성을 가지는 이차 대사 산물에 대한 본 발명자들의 지속적인 연구 결과 항균 활성 및 고지혈증에 치료효과를 보이는 새로운 지방산을 생성하는 해양 바실러스 (Marine Bacillus)속 미생물을 찾아내어 본 발명을 완성하게 된 것으로서, 본 발명이 해결하고자는 기술적 과제는 다양한 생리활성을 보이는 신규 물질을 생성하는 해양 미생물, 이 해양 미생물로부터 생성된 신규 물질 및 이 신규 물질의 제조방법을 제공하는 것이다. As a result of continuous research by the present inventors to solve the above problems, it was confirmed that marine microorganisms separated from the surface of marine sediment, marine algae and invertebrates produce secondary metabolites that exhibit physiological activities such as antimicrobial activity. Our ongoing research on secondary metabolites with physiological activity from marine microorganisms has shown that marine Bacillus produces new fatty acids that have therapeutic effects on antimicrobial activity and hyperlipidemia. The present invention is to solve the present invention by finding a microorganism of the genus Bacillus , the technical problem to be solved by the present invention is a marine microorganism to produce a new material showing a variety of physiological activity, a new material generated from the marine microorganism and the new material It is to provide a manufacturing method.

상기 기술적 과제를 달성하기 위하여 본 발명은 하기 화학식 1 ~ 4로 표시되는 화합물을 생성하는 기탁번호 KCTC 11975BP로 기탁된 해양 바실러스 속 미생물을 제공한다.In order to achieve the above technical problem, the present invention provides a microorganism of the genus Marine Bacillus deposited with accession number KCTC 11975BP to produce a compound represented by the following formula (1).

Figure 112013110654443-pat00001
Figure 112013110654443-pat00001

상기 화학식 1에서 R1=R2=H 임.In Formula 1, R 1 = R 2 = H.

Figure 112013110654443-pat00002
Figure 112013110654443-pat00002

상기 화학식 2에서 R=H 임.In Formula 2, R = H.

Figure 112013110654443-pat00003
Figure 112013110654443-pat00003

상기 화학식 3에서 R1=R2=R3=H 임.In Formula 3 R 1 = R 2 = R 3 = H.

Figure 112013110654443-pat00004
Figure 112013110654443-pat00004

상기 화학식 4에서 R1=R2=H 임.In Formula 4, R 1 = R 2 = H.

다른 기술적 과제를 달성하기 위하여, 본 발명은 화학식 1 ~ 4로 표시되는 지방산 화합물을 제공하며, 이 화합물들은 항균 활성을 가지는 것을 특징으로 한다.In order to achieve another technical problem, the present invention provides a fatty acid compound represented by the formula 1 to 4, characterized in that the compounds have an antimicrobial activity.

또 다른 기술적 과제를 달성하기 위하여, 본 발명은 상기 화학식 1 ~ 4로 표시되는 화합물의 제조방법을 제공하는데, 이 본 발명에 따른 제조방법은In order to achieve another technical problem, the present invention provides a method for preparing a compound represented by Formula 1 to 4, the production method according to the present invention

기탁번호 KCTC 11975BP로 기탁된 해양 바실러스 속 미생물을 0.1% 효모 추출물, 0.1% 비프 추출물, 0.2% 트립톤, 1% 덱스트로즈, 염도 12 g/L, pH 7.6의 변형된 베넷 배지에서 배양하는 단계; 및Cultivating the marine bacillus microorganisms deposited with accession no. ; And

상기 배양물을 원심분리한 상등액을 에틸아세테이트(EtOAc)로 추출한 후 에틸아세테이트 층을 증발 건조한 후 잔류 현탁물을 컬럼크로마토그래피를 이용하여 분획하고 분획물로부터 상기 화학식 1 ~ 4로 표시되는 화합물을 정제하는 단계를 포함한다.After extracting the culture supernatant centrifuged with ethyl acetate (EtOAc), and then the ethyl acetate layer was evaporated to dryness, the remaining suspension was fractionated by column chromatography and purified from the fractions the compound represented by the formula (1-4) Steps.

본 발명은 항균 활성을 보이며 고지혈증의 치료제로도 사용할 수 있는 상기 화학식 1 ~ 4로 표시되는 지방산 화합물을 제공하며, 이 지방산 화합물을 효율적으로 생산할 수 있는 해양 미생물 및 염도 12 g/L로 조절된 배지를 이용한 제조방법을 제공한다. 이와 같은 발명에 의하면 다양한 생리활성을 보이는 신규의 지방산 화합물 및 이의 제조방법에 제공되어 항생제 내성 병원균에 의한 위협, 항진균 약물의 제한적 효능 및 최근의 바이오테러리즘의 등장에 맞설 신규 항미생물 제제의 상업적 공급이 가능할 수 있다.The present invention provides a fatty acid compound represented by the above formulas (1) to (4) showing antibacterial activity and can also be used as a therapeutic agent for hyperlipidemia, and a medium controlled to 12 g / L of marine microorganisms and salinity that can efficiently produce the fatty acid compound It provides a manufacturing method using. According to the present invention, a novel fatty acid compound exhibiting various physiological activities and a method for preparing the same provide a commercial supply of a novel antimicrobial agent against the threat of antibiotic resistant pathogens, the limited efficacy of antifungal drugs, and the recent emergence of bioterrorism. It may be possible.

도 1은 09ID194 균주는 변형된 베넷 한천배지에서 잘 발달된 붉은 콜로니에 대한 사진이다.
도 2는 09ID194 균주의 계통도이다.
도 3는 화합물 1 ~ 2의 NMR 데이터 도면이다.
도 4은 화합물 1 ~ 4의 1H-1H COSY 및 HMBC 상과관계를 도시한 도면이다.
도 5는 화합물 1 ~ 4의 주요 ROSEY 상관관계를 도시한 도면이다.
도 6는 키시(Kishi)의 유니버설 NMR 데이터베이스 (database 2)에 기반한 화합물 1의 3,5-디올의 상대적 배열의 결정을 도시한 도면이다.
도 7은 화합물 1 ~ 4의 MTPA 에스테르에 대한 ΔδH 값 [ΔδH= δS - δR]을 나타낸 도면이다.
도 8은 화합물 3 ~ 4의 NMR 데이터 도면이다.Figure 1 is a 09ID194 strain is a picture of a well developed red colony in modified Bennett agar medium.
2 is a schematic diagram of a 09ID194 strain.
3 is a diagram of NMR data of Compounds 1 and 2. FIG.
Figure 4 is a diagram showing the relationship between phase and 1 H- 1 H and HMBC COSY of compound 1-4.
FIG. 5 shows the major ROSEY correlations for Compounds 1-4.
FIG. 6 shows the determination of the relative arrangement of 3,5-diols of compound 1 based on Kishi's Universal NMR database (Database 2).
FIG. 7 is a graph showing the Δδ H value [Δδ H = δ S − δ R ] for the MTPA esters of Compounds 1-4. FIG.
8 is a diagram of NMR data of Compounds 3-4.

이하 본 발명을 실시하기 위한 구체적인 내용을 균주의 분리 및 분류, 대량 배양을 통한 화합물을 정제 및 생리 활성 시험의 순서로 설명하고자 한다.Hereinafter, specific details for carrying out the present invention will be described in the order of isolation and classification of strains, purification of compounds through mass cultivation, and physiological activity tests.

<균주 09 ID194 의 분리 및 분류> < Isolation and classification of strain 09 ID194 >

균주 09ID194는 2009년 탐사에서 대한민국 남해의 이어도에서 채집한 퇴적물 샘플로부터 분리하였다. Strain 09ID194 was isolated from sediment samples collected in the island of the South Sea of South Korea during exploration in 2009.

분리과정을 설명하면, 퇴적물 샘플 1g을 무균 조건에서 멸균된 해수에 10-1, 10-2, 10- 3및 10-4로 희석시키고, 각 희석액 100㎕를 변형된 베넷 한천 배지(0.1% 효모 추출물, 0.1% 비프 추출물, 0.2% 트립톤, 1% 덱스트로즈, 100% 천연 해수, 1.8% 한천, 멸균전 7.2로 pH 조절)에 도말한 후 30℃에서 14일간 배양하여 생성된 균주 09ID194의 콜로니를 분리하였다. Referring to the separation process, 10 -1, 10 -2, 10 to deposit the sample on 1g of sterile water in a sterile condition - 3 and diluted to 10-4 and, (0.1% of the Bennett agar transform each dilution 100㎕ yeast Extract, 0.1% beef extract, 0.2% tryptone, 1% dextrose, 100% natural seawater, 1.8% agar, pH adjusted to 7.2 before sterilization) and then cultured at 30 ° C. for 14 days Colonies were separated.

상기의 변형된 베넷 한천 배지상에서 분리한 콜로니를 배양하였다. 09ID194 균주는 변형된 베넷 한천배지에서 잘 발달된 붉은 콜로니를 형성하였으며, 이에 대한 사진을 도 1로 첨부하였다. Colonies isolated on the modified Bennett agar medium were cultured. The 09ID194 strain formed well-developed red colonies in the modified Bennett agar medium, and a photograph thereof was attached to FIG. 1.

상기 균주는 16S rDNA 서열분석에 의하여 Bacillus sp.로 동정되었다. 한국생명과학연구원 생명자원센터에 KCTC 11975BP로 기탁되어 있다. 기탁된 균주에 대한 계통도를 도 2로 첨부하였다.The strain was identified as Bacillus sp. By 16S rDNA sequencing. It is deposited as KCTC 11975BP at Korea Life Science Research Institute. A schematic of the deposited strains is attached in FIG. 2.

<균주 09 ID194 대량 배양> < Strain 09 ID194 Mass Culture>

균주 09ID194 대량 배양은 변형된 베넷 배지(0.1% 효모 추출물, 0.1% 비프 추출물, 0.2% 트립톤, 1% 덱스트로즈, 염도 12 g/L, pH 7.6)에서 실시되었다. 상기 배지 200㎖를 500㎖들이 코니칼 플라스크에 분배하고 멸균하였다. Mass culture of strain 09ID194 was carried out in modified Bennett medium (0.1% yeast extract, 0.1% beef extract, 0.2% tryptone, 1% dextrose, salinity 12 g / L, pH 7.6). 200 ml of the medium was dispensed into 500 ml conical flasks and sterilized.

상기 분리과정에서 얻은 09ID194 균주의 단일 콜로니를 상기 플라스크에 무균적으로 접종시키고 24℃에서 2일간 120rpm의 회전 진탕배양기 상에서 배양하였다. A single colony of 09ID194 strain obtained in the isolation process was aseptically inoculated into the flask and incubated on a rotary shaker at 120 rpm for 2 days at 24 ° C.

상기 배양물을 0.2% v/v가 되도록 1ℓ멸균 배양 배지를 함유하는 2ℓ들이 플라스크 총 100개 플라스크에 접종하여 동일한 조건하에서 7일간 배양한 후 수집하였다.The cultures were inoculated in 100 flasks containing a total of 1 liter sterile culture medium to 0.2% v / v in 100 flasks were incubated for 7 days under the same conditions and collected.

<화합물의 추출 및 분리>Extraction and Separation of Compounds

상기 대량 배양에서 얻은 배양액 100ℓ를 원심분리하고 상등액을 에틸아세테이트(EtOAc, 2 x 100ℓ)로 추출한 후 40℃에서 회전 증발기를 사용하여 EtOAc 층을 건조 농축시켰다. 100 l of the culture solution obtained in the mass culture was centrifuged, the supernatant was extracted with ethyl acetate (EtOAc, 2 x 100 l), and the EtOAc layer was concentrated to dryness using a rotary evaporator at 40 ° C.

이어서 잔류 현탁액 20 g을 ODS open 칼럼크로마토그래피에 주입한 후, 이동상으로서 MeOH/H2O (v/v) (1:4, 2:3, 3:2, 4:1 및 100:0)을 사용하여 단계적 분리하였다. 20 g of the residual suspension was then injected into ODS open column chromatography, followed by MeOH / H 2 O (v / v) (1: 4, 2: 3, 3: 2, 4: 1 and 100: 0) as mobile phase. Stepwise separation.

MeOH/H2O (3:2, v/v)로 분리된 분획물을 semi-preparative ODS HPLC (50% MeOH 함유 H2O; 유속: 1.5 mL/min; 탐지기: RI)를 이용하여 9개의 분획물 F-1~ F-9를 얻었다. The fractions separated by MeOH / H 2 O (3: 2, v / v) were separated into nine fractions using semi-preparative ODS HPLC (H 2 O with 50% MeOH; flow rate: 1.5 mL / min; detector: RI). F-1 to F-9 were obtained.

13%MeOH-EtOAc; 5% MeOH-CHCl3; 10% Me0H-CHCl3; 6% MeOH-CHCl3를 이용한 isocratic 프로그램을 이용하여 순상 semi-preparative HPLC (유속: 1.3 mL/min; 탐지기: UV)로 정제하여 F-8, F-6, F-3 및 F-4로부터 상기 화학식 1 ~ 4로 표시되는 화합물을 순수한 상태로 화학식 1 화합물 3.4mg, 화학식 2 화합물 3.4mg, 화학식 3 화합물 3.3mg, 화학식 4 화합물 5.5mg를 얻었다. 13% MeOH-EtOAc; 5% MeOH-CHCl 3 ; 10% Me0H-CHCl 3 ; Purified by normal phase semi-preparative HPLC (flow rate: 1.3 mL / min; detector: UV) using an isocratic program with 6% MeOH-CHCl 3 , from F-8, F-6, F-3 and F-4. 3.4 mg of the compound of formula 1, 3.4 mg of the compound of formula 2, 3.3 mg of the compound of formula 3, and 5.5 mg of the compound of formula 4 were obtained in a pure state of the compound represented by the formulas 1 to 4.

하기 화학식 1로 표시되는 화합물(화합물 1)을 이어도마이신 A(Ieodomycin A), 화학식 2로 표시되는 화합물(화합물 2)을 이어도마이신 B(Ieodomycin B), 화학식 3으로 표시되는 화합물(화합물 3)을 이어도마이신 C(Ieodomycin C), 화학식 4로 표시되는 화합물(화합물 4)을 이어도마이신 D(Ieodomycin D)로 명명하였다.
Following the compound represented by the formula (1) (Ieodomycin A), followed by the compound represented by the formula (2) (Ieodomycin B), the compound represented by the formula (3) Yiodomycin C (Ieodomycin C), a compound represented by the formula (compound 4) was named as yiodomycin D (Ieodomycin D).

<< 이어도마이신Iodomycin A, 화합물 1 구조결정> A, Compound 1 Structure Determination>

이어도마이신 A는 노란색의 무정형 고체로서 분리되었다. HRESIMS 및 1D와 2D NMR 등의 데이터를 종합적으로 분석함으로써 분자식은 C13H22O4으로 결정되었다. 이어도마이신 A의 UV 분광스펙트럼은 λmax 232 nm에서 흡광 밴드를 보이는데, 이는 공역된 디엔(diene)에 해당되는 것을 알 수 있었다. 3400 및 1715 cm- 1 에서의 IR 흡광도는 각각 수산기 (OH) 및 에스테르 카르보닐기 (C=O)의 존재를 나타내었다. 13C NMR 및 HSQC 분광 스펙트럼에서 한 개의 특이한 말단 올레핀성 메틸렌 탄소 (δc 115.1)를 포함한 4 개의 올레핀 탄소 신호 (δc 115.1 -140.1), 4 개의 메틸렌 탄소 (δc 37.0 - 45.3), 2 개 산소와 결합된 메틴 탄소 (δc 66.6 및 68.8), 두개의 메틸 탄소(δc 16.8 및 52.1)(그 중의 하나는 산소와 결합됨)및 하나의 에스테르 카르보닐 탄소 (δc 174.0)가 존재함을 알 수 있었다(도 3 참조). 1H-1H COSY 스펙트럼을 분석한 결과, 2개의 분리된 스핀 시스템을 보였다; 하나는 H2-2 (δH 2.47)부터 H2-7 (δH 2.10/2.19)까지 이고, 나머지 하나는 H-9 (δH 5.86)부터 H2-11(δH 4.93/5.04)까지 이며, 이들의 C-8 (δc 140.1)과의 연결은 H2-7의 C-8과 C-9 (δc 127.0)과의 long-range correlations에 의해 결정되었다(도 4 참조). 메톡시(OCH3)기 (δH 3.67, s)와 카르보닐 탄소 C-1 (δc 174.0)과의 연결은, 메톡시 그룹의 양성자들이 C-1과 HMBC correlation을 나타내었기 때문에 쉽게 결정되었다 (도 4 참조). 유사하게, 나머지 메틸기 신호(δH 1.75, s, H3-12)는 C-7, C-8 및 C-9과 HMBC 교차 피크를 나타냈고, 이러한 연결관계로부터 C-8과의 연결이 결정되었다 (도 4 참조). 이러한 연관관계의 데이터로부터, 화합물 1의 평면 구조가 결정되었다. Iodomycin A was isolated as a yellow amorphous solid. The molecular formula was determined by C 13 H 22 O 4 by comprehensive analysis of data such as HRESIMS and 1D and 2D NMR. The UV spectral spectrum of the doomycin A shows an absorption band at λmax 232 nm, which corresponds to the conjugated diene. 3400 and 1715 cm - IR absorption at 1 showed the presence of each hydroxyl group (OH) and an ester group (C = O). Four olefin carbon signals (δc 115.1 -140.1) with one specific terminal olefinic methylene carbon (δc 115.1), four methylene carbons (δc 37.0-45.3), two oxygens in 13 C NMR and HSQC spectral spectra Methine carbon (δc 66.6 and 68.8), two methyl carbons (δc 16.8 and 52.1), one of which is bonded to oxygen, and one ester carbonyl carbon (δc 174.0). 3). 1 H- 1 H COSY Analysis of the spectrum showed two separate spin system; One is from H 2 -2 (δ H 2.47) to H 2 -7 (δ H 2.10 / 2.19) and the other is from H-9 (δ H 5.86) to H 2 -11 (δ H 4.93 / 5.04) and, in connection with these C-8 (δc 140.1) it was determined by long-range correlations between C-8 and C-9 (δc 127.0) of H 2 -7 (see Fig. 4). The linkage between the methoxy (OCH 3 ) group (δ H 3.67, s) and the carbonyl carbon C-1 (δc 174.0) was readily determined since the protons of the methoxy group showed C-1 and HMBC correlation ( See FIG. 4). Similarly, the remaining methyl group signals (δ H 1.75, s, H 3 -12) exhibited C-7, C-8 and C-9 and HMBC cross peaks, from which the association with C-8 was determined. (See FIG. 4). From this correlation data, the planar structure of compound 1 was determined.

ROESY correlation은 H-10과 H3-12 사이 및 H-9와 H-11b(δH 4.93) 사이에서는 관측되었지만, H-9와 H3-12 사이에는 아무 연관관계가 관측되지 않았다(도 5 참조). 이러한 연관관계로부터, C-8/C-9의 삼중치환된 이중 결합의 배열형태가 E로 결정되었다. ROESY correlation is not observed, there was no affinity between H-10 and H 3 -12, and between H-9 and H-11b (δ H 4.93) was observed between the, H-9 and H 3 -12 (5 Reference). From this association, the configuration of C-8 / C-9 triple substituted double bonds was determined as E.

화합물 1의 3,5-디올의 상대적 배열은 키시(Kishi)의 유니버셜 NMR 데이터베이스(Database 2)를 이용하여 정하였다. C-3/C-5의 13C NMR 공명은 1,3-디올 모델 시스템의 anti 배열과 잘 일치되었다(도 6 참조). 화합물 1의 3,5-디올의 절대 배열은 변형된 Mosher의 에스테르 방법으로 정하였다. The relative arrangement of 3,5-diol of compound 1 was determined using Kishi's Universal NMR Database (Database 2). The 13 C NMR resonance of C-3 / C-5 was in good agreement with the anti configuration of the 1,3-diol model system (see FIG. 6). The absolute configuration of 3,5-diol of compound 1 was determined by the modified Mosher ester method.

화합물 1을 (R)-(-)- 및 (S)-(+)-α-메톡시-α(트리플루오로메틸)페닐아세틸 클로라이드 (MTPA-Cl)를 함유한 무수 피리딘으로 각각 처리하여 각각 비스-(S)- 및 (R)- MTPA 에스테르 유도체 1a 및 1b를 얻었다. 상기의 두 개의 디에스테르 유도체의 모든 양성자 신호는 1H-1H COSY 실험으로 정하여졌고, 비스-(R)-MTPA 에스테르 (1b)의 1H 화학 전이값을 비스-(S)-MTPA 에스테르 (1a)의 상응하는 값에서 빼주었다. ΔδH 값 [ΔδH= δS - δR]은 도면 6에 나타내었다. H-3 (-0.11), H-4 (-0.12/-0.13) 및 H-5 (-0.17)의 음의 ΔδH 값은 리구에라(Riguera)에 의해 보고된 anti-1,3-디올류의 디에스테르류에 대한 ΔδH 패턴에 상응하는 것으로, 화합물 1의 C-3 및 C-5의 절대 배열이 각각 S 및 R이라는 것을 나타낸다. 따라서 화합물 1의 구조는 결론적으로 (3S,5R,8E)-메틸 3,5-디하이드록시-8-메틸운데카-8,10-디에노에이트라고 결정되었다.
Compound 1 was each treated with anhydrous pyridine containing ( R )-(-)-and ( S )-(+)-α-methoxy-α (trifluoromethyl) phenylacetyl chloride (MTPA-Cl), respectively. Bis- ( S )-and ( R ) -MTPA ester derivatives 1a and 1b were obtained. All proton signals are 1 H- 1 H COSY experiment was appointed to, bis of the two di-ester derivative - (R) -MTPA-bis the 1 H chemical transformation value of the ester (1b) - (S) -MTPA ester ( Subtracted from the corresponding value of 1a). The Δδ H value [Δδ H = δ S − δ R ] is shown in FIG. 6. H-3 (-0.11), H -4 (-0.12 / -0.13) and H-5 um Δδ H values of (-0.17) is Liguria Era (Riguera) an anti -1,3- diols reported by Corresponding to the Δδ H pattern for the diesters of, indicating that the absolute arrangement of C-3 and C-5 of Compound 1 is S and R , respectively. Therefore, the structure of compound 1 is conclusion (3 S, 5 R, 8 E) - was determined to be methyl 3,5-dihydroxy-8-methyl-undeca-diethoxy -8,10- no benzoate.

<< 이어도마이신Iodomycin B, 화합물 2 구조결정> B, Compound 2 Structure Determination>

이어도마이신 B는 광학 활성이 있는 백색의 무정형 고체로 분리되었다. 화합물 2의 분자식은 HRESIMS를 분석한 결과, m/z 233.1145에서 분자 이온 피크 ([M + Na]+)를 나타내어 C12H18O3 (화합물 1보다 탄소 1개, 수소 4개 및 산소 1개가 적음)로 결정되었다. 이의 IR 스펙트럼으로 수산기 (3365 cm-1) 및 δ-락톤기 (1715 cm-1)은 존재를 알 수 있었다. 이의 UV, IR, 1H 및 13C NMR 데이터(도 3 참조)와 화합물 1의 데이터를 비교하면, 화합물 2가 화합물 1과 유사한 골격을 가지는 것을 알 수 있었다. 화합물 2에서는 화합물 1의 C-1에 부착된 메톡시(OCH3)기가 없으며, 대신에, C-1이 C-5의 산소에 연결되어 락톤 고리를 형성하는 것이 달랐다. Iodomycin B was isolated as a white amorphous solid with optical activity. The molecular formula of compound 2 was analyzed by HRESIMS and showed a molecular ion peak ([M + Na] + ) at m / z 233.1145, indicating that C 12 H 18 O 3 (1 carbon, 4 hydrogen and 1 oxygen than compound 1) Less). Its IR spectrum revealed the presence of hydroxyl groups (3365 cm -1 ) and δ-lactone groups (1715 cm -1 ). Comparing the UV, IR, 1H and 13 C NMR data (see FIG. 3) with the data of Compound 1, it was found that Compound 2 had a skeleton similar to that of Compound 1. In compound 2, there was no methoxy (OCH 3 ) group attached to C-1 of compound 1, and instead, C-1 was connected to oxygen of C-5 to form a lactone ring.

UV, IR, 1D 및 2D NMR (도 3 참조) 데이터를 상세 분석하여, 화합물 2의 평면 구조를 명확하게 결정하였다. 화합물 2의 상대 배열은 3 J H -H 커플링 상수 및 ROESY 분광분석 데이터에 기초하여 결정되었다. H-10이 H3-12과의 ROE 상관관계를 가지는 것뿐만 아니라 H-9이 H-11b과의 ROE 상관관계를 가지는 것과, H-9와 H-10 사이의 아무런 ROE 상관관계가 없는 것으로부터 C-8/C-9 이중 결합이 E 기하배열을 가지고 있는 것을 알 수 있었다(도 5 참조). δ-락톤 고리의 H2-2의 분할 패턴 (splitting pattern) 및 화학적 전이값들 (δH 2.36, dd, J = 16.8, 7.3, H-2ax, δH 2.86, dd, J = 16.8, 1.3, H-2eq)로부터 C-3와 C-5의 anti 상대 배열을 명확하게 알 수 있었다. H-2ax 및 H-3ax 사이의 큰 인접 커플링 상수 (J = 7.3 Hz)는 3-OH 기가 equatorial 위치에 있다는 것을 나타내며, 이는 H-2eq 및 H-3ax간의 ROESY 상관관계에 의해 확인되었다. Trans-diaxial 커플링 (J = 10.4)이 H-3ax과 H-4ax 사이에 관찰되었고, 이 또한 3-OH기가 equatorial 위치를 가지는 것을 나타내었다. H-5의 axial 위치는 H-4ax (J = 7.3) 및 H-6ax (J =10.4)과의 커플링 상수에 의해 알 수 있었고, H-4eq와 H-5ax와의 ROESY 상관관계에 의해 확인되었다(도 5 참조). H-2ax와 H-4ax 사이 및 H-3ax와 H-5ax 사이의 ROESY 상관관계가 이러한 사실을 뒷받침하였다. 이러한 상관관계 데이터로부터 δ-락톤 고리가 의자 형태를 취하고 있음을 알 수 있었다. 화합물 2의 C-3의 절대적 배열은 화합물 1에서와 같이 변형된 Mosher의 방법을 적용함으로써 결정되었다. 화합물 2를 (R)-(-)- 및 (S)-(+)-MTPA-Cl으로 아실화하여 각각 모노-(S)- 및 (R)-MTPA 에스테르 (2a 및 2b)를 만들었다. 이러한 MTPA 에스테르류에 대한 1H NMR 데이터(도 3 참조)를 분석하여 C-3에서의 절대 배열이 S 임을 나타내었다. UV, IR, 1D and 2D NMR (see FIG. 3) data were analyzed in detail to clearly determine the planar structure of Compound 2. The relative arrangement of compound 2 was determined based on the 3 J H -H coupling constants and ROESY spectroscopic data. H-10 that is not yet ROE correlations between H 3 as well as with the -12 and the ROE correlations are H-9 having the ROE correlations between H-11b as, H-9 and H-10 It can be seen that the C-8 / C-9 double bond has an E geometry (see FIG. 5). Splitting pattern and chemical transition values of H 2 -2 of the δ-lactone ring (δ H 2.36, dd, J = 16.8, 7.3, H-2ax, δ H 2.86, dd, J = 16.8, 1.3, H-2eq) clearly shows the anti- relative arrangement of C-3 and C-5. The large adjacent coupling constant ( J = 7.3 Hz) between H-2ax and H-3ax indicates that the 3-OH group is in the equatorial position, which was confirmed by the ROESY correlation between H-2eq and H-3ax. Trans- diaxial coupling ( J = 10.4) was observed between H-3ax and H-4ax, which also showed that the 3-OH group had an equatorial position. The axial position of H-5 was determined by the coupling constants of H-4ax ( J = 7.3) and H-6ax ( J = 10.4) and confirmed by the ROESY correlation between H-4eq and H-5ax. (See Figure 5). The ROESY correlation between H-2ax and H-4ax and between H-3ax and H-5ax supported this fact. From these correlation data, it can be seen that the δ-lactone ring takes the form of a chair. The absolute arrangement of C-3 of Compound 2 was determined by applying the modified Mosher's method as in Compound 1. Compound 2 (R) - (-) - and (S) - (+) - MTPA-Cl by acylating the respective mono- made the MTPA ester (2a and 2b) - (S) - and (R). 1 H NMR data (see FIG. 3) for these MTPA esters were analyzed to show that the absolute configuration at C-3 is S.

화합물 2에서 C-5의 배열은 C-3와 C-5 사이의 anti 관계를 기반으로 하여 R 로서 결정되었고, 이는 커플링 상수 및 ROESY 상관관계에 의해 알 수 있었다. 광범위한 1D 및 2D 분광자료 분석으로 화합물 2의 구조를 (3S,5R,8E)-3-hydroxy-5-(8-methylhexa-8,10-dienyl)tetrahydro-1H-pyran-1-one으로 결정하였다.
The arrangement of C-5 in Compound 2 was determined as R based on the anti- relationship between C-3 and C-5, which can be seen by the coupling constant and ROESY correlation. Extensive analysis of 1D and 2D spectroscopy reveals the structure of Compound 2 as (3 S , 5 R , 8 E ) -3-hydroxy-5- (8-methylhexa-8,10-dienyl) tetrahydro-1 H -pyran-1- One decided.

<< 이어도마이신Iodomycin C, 화합물 3 구조결정> C, Compound 3 Structure Determination>

이어도마이신 C는 엷은 무정형 고체로서, HRESIMS (m/z 227.1288 [M - H]-) 및 1H과 13C NMR 데이터의 분석결과 분자식은 C12H20O4로 결정되었고 (도 8 참조), 불포화도 3을 가지고 있는 것을 알 수 있었다. 화합물 3은 IR 분광스펙트럼에서 3349 및 1711 cm- 1 에서 흡광 밴드를 나타내었는데, 이는 각각 수산기 (OH)와 카르보닐기 (C=O)에 해당되는 것이다. UV 분광스펙트럼에서 λmax 230 nm에서 최대 흡광을 보였고, 이는 공역된 디엔에 기인한 것이었다. 화합물 3의 수소 및 탄소의 시그널들 (도 8 참조)는 2D NMR 실험 (1H-1H COSY, HSQC 및 HMBC)에 의해 명확하게 결정되었다. Iodomycin C is a pale amorphous solid, HRESIMS ( m / z 227.1288 [M − H] − ) and analysis of 1 H and 13 C NMR data showed that the molecular formula was C 12 H 20 O 4 (see FIG. 8), It can be seen that the degree of unsaturation is 3. Compound 3 is 3349 and 1711 cm in the IR spectrum - eotneunde shows the absorption band at 1, which is applicable to each of the hydroxyl group (OH) and the carbonyl group (C = O). Maximum absorption at λmax 230 nm was shown on the UV spectroscopy, due to the conjugated diene. Signals of hydrogen and carbon of compound 3 (See FIG. 8) was clearly determined by 2D NMR experiments ( 1 H- 1 H COSY, HSQC and HMBC).

1H 및 13C NMR 스펙트럼(도 7)에서 잘 분리된 수소와 탄소 시그널들을 보여주었고, 화합물 3은 H2-2 (δH 2.45)로부터 H3-12(δH 1.13)까지의 단일 스핀 시스템을 가지고 있음을 1H-1H COSY 스펙트럼을 분석함으로서 알 수 있었다(도 4 참조). C-4/C-5 및 C-6/C-7의 이중 결합은 둘 다는 E geometry를 가지며, 이는 H-4와 H-5 사이(J = 15.3 Hz) 및 H-6과 H-7 사이(J = 15.3 Hz)의 큰 스칼라 커플링 상수에 의해 확인되었다. 이것은 또한 H-4와 H-6 사이 및 H-5와 H-7 사이의 ROESY 상관관계에 의해 확인되었다 (도 5 참조). 화합물 1과 2와 같이, 화합물 3a를 Mosher의 디에스테르 (S)-MTPA (3b) 및 (R)-MTPA (3c)로 전환하고, 1H-NMR 데이터를 분석한 결과, 화합물 3의 C-3 및 C-11는 각각 S 및 R 절대 배열을 가지고 있음을 알 수 있었다. 결론적으로 화합물 3의 구조는 (3S,4E,6E,11R)-3,11-dihydroxydodeca-4,6-dienoic acid 로 결정되었다.
 1 H and 13 C NMR spectra (FIG. 7) showed well separated hydrogen and carbon signals, compound 3 being a single spin system from H 2 -2 (δ H 2.45) to H 3 -12 (δ H 1.13) a it was found that by the 1 H- 1 H COSY analyzing the spectrum has (see Fig. 4). Double bonds of C-4 / C-5 and C-6 / C-7 both have E geometry, which is between H-4 and H-5 ( J = 15.3 Hz) and between H-6 and H-7 It was confirmed by a large scalar coupling constant of ( J = 15.3 Hz). This was also confirmed by the ROESY correlation between H-4 and H-6 and between H-5 and H-7 (see FIG. 5). Like compound 1 and 2, compound 3a Conversion to Mosher diesters ( S ) -MTPA (3b) and ( R ) -MTPA (3c) and analysis of 1 H-NMR data showed that C-3 and C-11 of compound 3 were S and R, respectively. I could see that it has an absolute array. In conclusion, the structure of compound 3 was determined to be (3S, 4E, 6E, 11R) -3,11-dihydroxydodeca-4,6-dienoic acid.

<< 이어도마이신Iodomycin D, 화합물 4 구조결정> D, compound 4 structure determination>

이어도마이신 D는 광학 활성을 가지는 밝은 황색의 무정형 고체로 분리되었다. 화합물 4의 분자식은 HRESIMS 및 13C NMR 데이터에 의해 C10H16O3로 결정되었고, 화합물 4는 화합물 3보다 탄소 2개, 수소 4개 및 탄소 1개가 적었고, 동일한 불포화도를 가지고 있었다. Iodomycin D was isolated as a light yellow amorphous solid with optical activity. The molecular formula of compound 4 was determined to be C 10 H 16 O 3 by HRESIMS and 13 C NMR data, and compound 4 had 2 carbons, 4 hydrogens and 1 carbon less than compound 3, and had the same degree of unsaturation.

화합물 4의 UV 및 IR 스펙트럼은 화합물 3의 것과 매우 유사하였으며, 이들 스펙트럼으로부터 카르보닐기 (1684 cm-1), 수산기 (3365 cm-1) 및 공역 디엔 (λmax 254 nm)의 존재를 알 수 있었다. 도 8에 나타난 바와 같이, 수소와 결합된 모든 탄소를 HSQC 스펙트럼 분석으로 알 수 있었다. 화합물 4는 H-2 (δH 5.78)로부터 시작하여 H3-10 (δH 1.14)까지 끝나는 오직 하나의 스핀 시스템을 가지고 있음을 1H-1H COSY 데이터로부터 알 수 있었다. α, β, γ, δ의 불포화 카르보닐 moiety는, H-2 (δH 5.78) 및 H-4 (δH 6.24)가 각각 C-1 (δc 171.5) 및 C-3 (δc 146.5)와 long-range correlation을 보이는 것으로부터 알 수 있었다. C-2/C-3 및 C-4/C-5 이중결합의 E 기하배열은 H-2와 H-3 (J = 15.3 Hz) 사이, 및 H-4와 H-5 (J =15.3 Hz) 사이의 큰 커플링 상수로부터 알 수 있었고(도 8 참조), ROESY 상관관계에 의해 확인되었다 (도 5 참조). 화합물 4의 입체센터 C-9의 절대 배열은 변형된 Mosher의 방법에 의해 결정되었다: 각각의 모노-(S)- 및 (R)-MTPA 에스테르를 준비하고 1H NMR 데이터 분석하였다. 화합물 4의 C-9의 R configuration은 가이드라인에 준하여 결정되었으며, 그 구조는 (2E,4E,9R)-9-hydroxydeca-2,4-dienoic acid (2E,4E,9R)-9-hydroxydeca-2,4-dienoic acid로 명확하게 결정되었다.The UV and IR spectra of compound 4 were very similar to those of compound 3, indicating the presence of carbonyl groups (1684 cm −1 ), hydroxyl groups (3365 cm −1 ) and conjugated dienes (λ max 254 nm). As shown in FIG. 8, all carbons bonded with hydrogen were identified by HSQC spectral analysis. Compound 4 It was found from the H-2 only that the 1 H- 1 H COSY data with a single spin system starting from (δ H 5.78) and the end to H 3 -10 (δ H 1.14) . Unsaturated carbonyl moieties of α, β, γ, and δ are H-2 (δ H 5.78) and H-4 (δ H 6.24), with C-1 (δc 171.5) and C-3 (δc 146.5) and long, respectively. This can be seen by showing the -range correlation. The E geometry of C-2 / C-3 and C-4 / C-5 double bonds is between H-2 and H-3 ( J = 15.3 Hz), and H-4 and H-5 ( J = 15.3 Hz ) From large coupling constants (see FIG. 8) and confirmed by ROESY correlation (see FIG. 5). The absolute configuration of stereocenter C-9 of compound 4 was determined by modified Mosher's method: Each mono- ( S )-and ( R ) -MTPA ester was prepared and analyzed by 1 H NMR data. The R configuration of C-9 of Compound 4 was determined according to the guidelines, and its structure was (2E, 4E, 9R) -9-hydroxydeca-2,4-dienoic acid (2E, 4E, 9R) -9-hydroxydeca- Clearly determined by 2,4-dienoic acid.

<화합물 1 ~ 4인 <Compound 1-4 이어도마이신Iodomycin A ~ D의 구조> Structure of A ~ D>

<화학식 1>&Lt; Formula 1 >

Figure 112013110654443-pat00005
Figure 112013110654443-pat00005

상기 화학식 1에서 R1=R2=H 임.In Formula 1, R 1 = R 2 = H.

<화학식 2>(2)

Figure 112013110654443-pat00006
Figure 112013110654443-pat00006

상기 화학식 2에서 R=H 임.In Formula 2, R = H.

<화학식 3><Formula 3>

Figure 112013110654443-pat00007
Figure 112013110654443-pat00007

상기 화학식 3에서 R1=R2=R3=H 임.In Formula 3 R 1 = R 2 = R 3 = H.

<화학식 4>&Lt; Formula 4 >

Figure 112013110654443-pat00008
Figure 112013110654443-pat00008

상기 화학식 4에서 R1=R2=H 임.In Formula 4, R 1 = R 2 = H.

<화합물 1 ~ 4의 생리 활성><Physiological activity of compounds 1 to 4>

-항균 활성-Antimicrobial Activity

화합물 1-4의 최소 저해 농도(MIC)를 통상적인 배양액 희석법을 사용하여 결정하였으며, 화합물 1-4를 세가지의 미생물 균주에 대하여 항균활성을 측정하였다: Bacillus subtilis(KCTC 1021), E. coli(KCTC 1923), 및 Saccharomyces cerevisiae (KCTC 7913). The minimum inhibitory concentration (MIC) of compound 1-4 was determined using conventional culture dilution method, and compound 1-4 was measured for antimicrobial activity against three microbial strains: Bacillus subtilis (KCTC 1021), E. coli ( KCTC 1923), and Saccharomyces cerevisiae (KCTC 7913).

항균 및 항효모 시험을 각각 영양 배양액 및 효모 말토즈 배양액에서 수행하였다. 각 화합물의 희석액을 0.5-512.0 ㎍/㎖의 범위에 걸쳐 96-마이크로타이터 플레이터에 마련하였다. 각 균주의 하루 배양액을 마련하여 균주의 최종 농도를 0.5 McFarland Standard와 배양액 탁도를 비교하여 1.5 x 108 cfu/mL로 조정하였다. 배양액 20㎕를 화합물 1-4의 각 희석액에 첨가하고 각 웰의 최종 부피를, 각자의 배양 배지를 사용하여 200㎕로 조정하고, 플레이트를 세균에 대해서는 24 시간동안 37℃에서 효모에 대해서은 48시간 동안 30℃에서 배양하였다. 최소 저해 농도 (MIC)는 미생물이 탁도의 존재에 의해 측정시 가시적인 성장을 나타내지 않는 샘플의 최소 농도이다.Antibacterial and anti yeast tests were performed in nutrient cultures and yeast maltose cultures, respectively. Dilutions of each compound were prepared in 96-microtiter plates over a range of 0.5-512.0 μg / ml. The daily culture of each strain was prepared and the final concentration of the strain was adjusted to 1.5 x 10 8 cfu / mL by comparing the culture medium turbidity with 0.5 McFarland Standard. 20 μl of culture was added to each dilution of compound 1-4 and the final volume of each well was adjusted to 200 μl using their respective culture medium, and the plate was allowed to cultivate for 24 hours for bacteria and 48 hours for yeast at 37 ° C. Incubated at 30 ° C. for 30 minutes. Minimum Inhibitory Concentration (MIC) is the minimum concentration of a sample in which microorganisms do not show visible growth as measured by the presence of turbidity.

화합물 1 ~ 4는 Bacillus subtilis 및 Escherichia coli에 대해서는 32-64 ㎍/㎖의 최소 저해 농도(MIC)로 항균활성을 보였으나, Saccharomyces cerevisiae에 대해서는 256㎍/㎖의 MIC로, 약한 성장저해 활성을 나타내었다.Compounds 1-4 are Bacillus subtilis and Escherichia For coli showed an antimicrobial activity with a minimum inhibitory concentration (MIC) of 32-64 ㎍ / ㎖ or, Saccharomyces For cerevisiae , 256 µg / ml MIC showed weak growth inhibition activity.

-고지혈증 치료 효과-Hyperlipidemia treatment effect

화합물 1 ~ 4는 스타틴 계열의 화합물과 구조가 유사하여 혈 중 콜레스테롤 치를 낮추어 고지혈증에 대한 치료 효과를 보일 것으로 예측된다.
Compounds 1 to 4 are similar in structure to statin compounds, and are expected to lower the cholesterol level in the blood to treat the hyperlipidemia.

<결론><Conclusion>

결론적으로, 4 가지 신규한 희귀 하이드록시 불포화 지방산인 화합물 1 ~ 4(이어도마이신 A-D)를 생성하였고, 이들 물질은 그람-양성 및 그람-음성 병원균에 대하여 항균 활성을 나타내었다. In conclusion, four novel rare hydroxyunsaturated fatty acids, Compounds 1-4 (Eytomycin A-D), were produced, which showed antimicrobial activity against Gram-positive and Gram-negative pathogens.

또한 화합물 1-4는 이러한 바실러스 종에 의해서 생성되지만, 낮은 염도 (12 g/L)에서만 생성하고, 높은 염도(32 g/L)의 배양 배지에서는 생성하지 않았다.In addition, compound 1-4 Produced by these Bacillus species, but produced only at low salinity (12 g / L) and not at high salinity (32 g / L) culture medium.

<참고><Reference>

비스Vis -(- ( SS )-) - MTPAMTPA 에스테르 (화합물 1a)의 제조  Preparation of Ester (Compound 1a)

화합물 1 (0.6 mg)을 200㎕ 피리딘에 녹이고 실온에서 10 분간 교반하였다. 1의 비스-(S)-MTPA 에스테르 (화합물 1a)를 제조하기 위해, 20㎕ (R)-(-)-MTPA-Cl을 반응 바이얼에 첨가하고 혼합물을 실온에서 16시간 교반하였다. 반응 완결은 LC/MS로 모니터하였다. 반응 혼합물을 진공에서 건조시키고, EtOAc에 다시 녹이고, H2O로 세척하고, 이동상으로 10% MeOH를 함유한 CHCl3 를 사용하여 실리카 컬럼으로 정제하여 1a을 수득하였다 (0.45 mg).Compound 1 (0.6 mg) was dissolved in 200 μl pyridine and stirred at room temperature for 10 minutes. To prepare bis- ( S ) -MTPA ester (Compound 1a) of 1, 20 μl ( R )-(-)-MTPA-Cl was added to the reaction vial and the mixture was stirred for 16 hours at room temperature. Reaction completion was monitored by LC / MS. The reaction mixture was dried in vacuo, taken up in EtOAc again, washed with H 2 O and purified by silica column with CHCl 3 containing 10% MeOH as mobile phase to give 1a (0.45 mg).

화합물 1a: 무정형 고체; 1H NMR 데이터 (CD3OD): δH 2.64 (H2-2, dd, J = 10.0, 6.5 Hz), 5.34 (H-3, m), 1.92 (H-4b, m), 1.94 (H-4a, m), 4.88 (H-5, m), 1.74 (H2-6, m), 1.91 (H-7b, m), 1.96 (H-7a,m), 5.73 (H-9, d, J = 10.5 Hz), 6.51, (H-10, ddd, J = 16.5, 10.5, 10.5 Hz), 4.96 (H-11b, d, J = 10.5 Hz), 5.04 (H-11a, d, J = 16.5 Hz), 1.67 (H3-12, s), 3.58 (OCH3, s), 3.54 (2 x OCH3, s) 7.38-7.64 (10H, m);ESIMS m/z 697.34 [M + Na]+.
 Compound 1a: amorphous solid; 1 H NMR data (CD 3 OD): δ H 2.64 (H2-2, dd, J = 10.0, 6.5 Hz), 5.34 (H-3, m), 1.92 (H-4b, m), 1.94 (H- 4a, m), 4.88 (H-5, m), 1.74 (H2-6, m), 1.91 (H-7b, m), 1.96 (H-7a, m), 5.73 (H-9, d, J = 10.5 Hz), 6.51, (H-10, ddd, J = 16.5, 10.5, 10.5 Hz), 4.96 (H-11b, d, J = 10.5 Hz), 5.04 (H-11a, d, J = 16.5 Hz ), 1.67 (H 3-12, s), 3.58 (OCH 3, s), 3.54 (2 × OCH 3 , s) 7.38-7.64 (10H, m); ESIMS m / z 697.34 [M + Na] + .

비스Vis -(- ( RR )-) - MTPAMTPA 에스테르 (화합물 1b)의 제조:  Preparation of Ester (Compound 1b):

전체적으로 화합물 1a와 유사한 방식으로, (S)-(+)-MTPA-Cl를 사용하여 비스-(R)-MTPA 에스테르 (화합물 1b)를 얻었다. In a similar manner to Compound 1a as a whole, bis- ( R ) -MTPA ester (Compound 1b) was obtained using ( S )-(+)-MTPA-Cl.

화합물 1b: 무정형 고체 (0.5 mg); 1H NMR 데이터 (CD3OD): δH 2.66 (H2-2, dd, J = 9.0, 6.5 Hz), 5.45 (H-3, m), 2.04 (H-4b, m), 2.07 (H-4a, m), 5.05 (H-5, m), 1.71 (H2-6, m), 1.90 (H-7b, m), 1.92 (H-7a, m), 5.72 (H-9, d, J = 10.0 Hz), 6.03, (H-10, ddd, J = 16.5, 10.0, 10.0 Hz), 4.96 (H-11b, d, J = 10.0 Hz), 5.04 (H-11a, d, J = 16.5 Hz), 1.67 (H3-12, s), 3.59 (OCH3, s), 3.60 (2 x OCH3, s) 7.44-7.57 (10H, m); ESIMS m/z 697.45 [M + Na]+. Compound 1b: amorphous solid (0.5 mg); 1 H NMR data (CD 3 OD): δ H 2.66 (H2-2, dd, J = 9.0, 6.5 Hz), 5.45 (H-3, m), 2.04 (H-4b, m), 2.07 (H- 4a, m), 5.05 (H-5, m), 1.71 (H2-6, m), 1.90 (H-7b, m), 1.92 (H-7a, m), 5.72 (H-9, d, J = 10.0 Hz), 6.03, (H-10, ddd, J = 16.5, 10.0, 10.0 Hz), 4.96 (H-11b, d, J = 10.0 Hz), 5.04 (H-11a, d, J = 16.5 Hz ), 1.67 (H3-12, s), 3.59 (OCH3, s), 3.60 (2 x OCH3, s) 7.44-7.57 (10H, m); ESIMS m / z 697.45 [M + Na] + .

모노-(Mono- ( SS )- 및 () - and ( RR )-) - MTPAMTPA 에스테르 (화합물 2a 및 2b)의 제조.  Preparation of Esters (Compounds 2a and 2b).

화합물 2a (0.5 mg) 및 2b (0.62 mg)를 각각 (R)-(-)- 및 (S)-(+)-MTPA-Cl로부터, 화합물 1a와 1b에 대해 전술한 바와 같이, 유사한 방식으로 제조하였다.Compounds 2a (0.5 mg) and 2b (0.62 mg) from ( R )-(-)-and ( S )-(+)-MTPA-Cl, respectively, in a similar manner as described above for compounds 1a and 1b Prepared.

화합물 2a: 무정형 고체; 1H NMR 데이터 (CD3OD): δH 2.53 (H-2b, dd, J = 16.8, 6.0), 2.06 (H-2a, dd, J = 16.8, 6.0), 5.40 (H-3, m), 2.50 (H-4b, m), 2.52 (H-4a, m), 4.41 (H-5, m), 1.80 (H2-6, m), 2.17 (H-7b, m), 2.26 (H-7a, m), 5.90 (H-9, d, J = 11.0), 6.59 (H-10, ddd, J = 16.5, 11.0, 11.0), 4.97 (H- Compound 2a: amorphous solid; 1 H NMR data (CD 3 OD): δ H 2.53 (H-2b, dd, J = 16.8, 6.0), 2.06 (H-2a, dd, J = 16.8, 6.0), 5.40 (H-3, m) , 2.50 (H-4b, m), 2.52 (H-4a, m), 4.41 (H-5, m), 1.80 (H2-6, m), 2.17 (H-7b, m), 2.26 (H- 7a, m), 5.90 (H-9, d, J = 11.0), 6.59 (H-10, ddd, J = 16.5, 11.0, 11.0), 4.97 (H-

11b, d, J = 11.0), 5.07 (H-11a, d, J = 16.5), 1.77 (H3-12, s), 3.53 (OCH3, s), 7.37-7.50 (5H, m); ESIMS m/z 449.24 [M + Na]+.11b, d, J = 11.0, 5.07 (H-11a, d, J = 16.5), 1.77 (H3-12, s), 3.53 (OCH 3 , s), 7.37-7.50 (5H, m); ESIMS m / z 449.24 [M + Na] + .

화합물 2b: 무정형 고체; 1H NMR 데이터 (CD3OD): δH 2.67 (H-2b, dd, J = 17.0, 5.5), 3.10 (H-2a, dd, J = 17.0, 5.5), 5.56 (H-3, m), 2.44 (H-4b, m), 2.46 (H-4a, m), 4.35 (H-5, m), 1.61 (H2-6, m), 2.16 (H-7b, m), 2.22 (H-7a, m), 5.88 (H-9, d, J = 10.5), 6.58 (H-10, ddd, J = 16.5, 10.5, 10.5), 4.97 (H-11b, d, J = 10.5), 5.07 (H-11a, d, J = 16.5), 1.77 (H3-12, s), 3.52 (OCH3, s), 7.40-7.57 (5H, m); ESIMS m/z 449.26 [M + Na]+. Compound 2b: amorphous solid; 1 H NMR data (CD3OD): δ H 2.67 (H-2b, dd, J = 17.0, 5.5), 3.10 (H-2a, dd, J = 17.0, 5.5), 5.56 (H-3, m), 2.44 (H-4b, m), 2.46 (H-4a, m), 4.35 (H-5, m), 1.61 (H2-6, m), 2.16 (H-7b, m), 2.22 (H-7a, m), 5.88 (H-9, d, J = 10.5), 6.58 (H-10, ddd, J = 16.5, 10.5, 10.5), 4.97 (H-11b, d, J = 10.5), 5.07 (H- 11a, d, J = 16.5), 1.77 (H3-12, s), 3.52 (OCH3, s), 7.40-7.57 (5H, m); ESIMS m / z 449.26 [M + Na] + .

화합물 3a의 제조. Preparation of Compound 3a.

화합물 3 (2.0 mg)을 1 mL 무수 MeOH에 녹이고,200㎕의 2 M (트리메틸실릴)디아조메탄을 용액에 첨가하였다. 반응 혼합물을 실온에서 교반하고, 반응 완결은 1 시간 내에 LC/MS 분석으로 확인하였다. 반응 혼합물을 진공 건조시키고, 65% MeOH을 함유하는 H2O을 사용하는 분석적 ODS HPLC로 정제하여 화합물 3의 메틸 에스테르 (화합물 3a, 1.8 mg)를 얻었다.Compound 3 (2.0 mg) was dissolved in 1 mL anhydrous MeOH and 200 μl of 2 M (trimethylsilyl) diazomethane was added to the solution. The reaction mixture was stirred at room temperature and reaction completion was confirmed by LC / MS analysis within 1 hour. The reaction mixture is dried in vacuo and purified by analytical ODS HPLC using H 2 O containing 65% MeOH to methyl ester of compound 3. (Compound 3a, 1.8 mg) was obtained.

화합물 3a: 엷은색, 무정형 고체; 1H NMR 데이터 (CD3OD): δH 2.50 (H2-2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.59 (H-4, dd, J = 15.3, 7.0 Hz), 6.22 (H-5, dd, J = 15.3, 10.3 Hz), 6.04 (H-6, dd, J = 15.3, 10.3 Hz), 5.71 (H-7, dt, J = 15.3, 7.0 Hz), 2.10 (H2-8, m), 1.38 (H-9b, m), 1.50 (H-9a, m), 1.42 (H2-10, m), 3.71 (H-11, m), 1.14 (H3-12, d, J = 6.5 Hz), 3.67 (OCH3, s); ESIMS m/z 241.33 [M - H]-. Compound 3a: pale, amorphous solid; 1 H NMR data (CD 3 OD): δ H 2.50 (H2-2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.59 (H-4, dd, J = 15.3, 7.0 Hz) , 6.22 (H-5, dd, J = 15.3, 10.3 Hz), 6.04 (H-6, dd, J = 15.3, 10.3 Hz), 5.71 (H-7, dt, J = 15.3, 7.0 Hz), 2.10 (H2-8, m), 1.38 (H-9b, m), 1.50 (H-9a, m), 1.42 (H 2 -10, m), 3.71 (H-11, m), 1.14 (H3-12 , d, J = 6.5 Hz), 3.67 (OCH 3 , s); ESIMS m / z 241.33 [M−H] − .

비스Vis -(- ( SS )-) - MTPAMTPA 에스테르(화합물 3b) 제조.  Preparation of Ester (Compound 3b).

화합물 3a (0.9 mg)를 1 mL 피리딘에 녹이고 실온에서 30분간 바이얼에서 교반하였다. 4-디메틸아미노피리딘 (DMAP)의 몇 개 결정을 첨가하고 용액을 30분간 더 교반하였다. 3a의 비스-(S)-MTPA 에스테르 (3b)의 제조를 위해, 10㎕(R)-(-)-MTPA-Cl를 첨가하고, 반응액을 실온에서 3 시간 교반하였다. 반응 완결은 LC/MS로 모니터하였다. 반응액를 200㎕ MeOH 첨가하여 냉각하였다. 반응 혼합물을 진공에서 건조시키고, H2O에 다시 녹이고, EtOAc (3 x 3 mL)로 추출하였다. 추출액을 67% MeOH을 함유한 H2O을 이동상으로 사용하는 ODS 분석 컬럼 상에서 정제하여 화합물 3b (0.7 mg)를 얻었다.Compound 3a (0.9 mg) was dissolved in 1 mL pyridine and stirred in a vial at room temperature for 30 minutes. Several crystals of 4-dimethylaminopyridine (DMAP) were added and the solution stirred for 30 more minutes. For the preparation of 3a bis- ( S ) -MTPA ester ( 3b ), 10 μl ( R )-(-)-MTPA-Cl was added and the reaction solution was stirred at room temperature for 3 hours. Reaction completion was monitored by LC / MS. The reaction solution was cooled by adding 200 µl MeOH. The reaction mixture was dried in vacuo, taken up in H 2 O again and extracted with EtOAc (3 × 3 mL). The extract was purified on an ODS analytical column using H 2 O containing 67% MeOH as the mobile phase to give compound 3b. (0.7 mg) was obtained.

화합물 3b: 엷은색, 무정형 고체; 1H NMR 데이터 (CD3OD): δH 3.01 (H2-2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.55 (H-4, dd, J = 15.0, 7.5 Hz), 6.20 (H-5, dd, J = 15.0, 10.0 Hz), 5.92 (H-6, dd, J = 15.0, 10.0 Hz), 5.56 (H-7, dt, J = 15.0, 6.0 Hz), 1.98 (H2-8, m), 1.35 (H-9b, m), 1.50 (H-9a, m), 1.48 (H2-10,m), 3.71 (H-11, m), 1.14 (H3-12, d, J = 6.5 Hz), 3.67 (OCH3, s), 3.54 (2 x OCH3, s), 7.35-7.60 (10H, m); ESIMS m/z 697.33 [M + Na]+. Compound 3b: pale, amorphous solid; 1 H NMR data (CD 3 OD): δ H 3.01 (H 2 -2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.55 (H-4, dd, J = 15.0, 7.5 Hz ), 6.20 (H-5, dd, J = 15.0, 10.0 Hz), 5.92 (H-6, dd, J = 15.0, 10.0 Hz), 5.56 (H-7, dt, J = 15.0, 6.0 Hz), 1.98 (H 2 -8, m), 1.35 (H-9b, m), 1.50 (H-9a, m), 1.48 (H 2 -10, m), 3.71 (H-11, m), 1.14 (H 3 -12, d, J = 6.5 Hz), 3.67 (OCH 3, s), 3.54 (2 x OCH 3, s), 7.35-7.60 (10H, m); ESIMS m / z 697.33 [M + Na] + .

비스Vis -(- ( RR )-) - MTPAMTPA 에스테르(화합물 3c) 제조.  Preparation of Ester (Compound 3c).

화합물 3a (0.9 mg)의 비스-(R)-MTPA 에스테르를 (S)-(+)-MTPA-Cl로부터 3b와 유사하게 제조하였다. 화합물 3a의 비스-(R)-MTPA 에스테르를 65% MeOH을 함유한 H2O를 용리액으로 사용하는 ODS 분석 컬럼상에서 정제하여 화합물 3c (0.6 mg)를 수득하였다.Bis- ( R ) -MTPA ester of compound 3a (0.9 mg) was prepared analogously from 3b from ( S )-(+)-MTPA-Cl. Of compound 3a Compound 3c by purifying bis- ( R ) -MTPA ester on an ODS analytical column using H 2 O containing 65% MeOH as eluent (0.6 mg) was obtained.

화합물 3c: 엷은색, 무정형 고체; 1H NMR 데이터 (CD3OD): δH 2.50 (H2-2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.85 (H-4, dd, J = 15.0, 8.5 Hz), 6.22 (H-5, dd, J = 15.0, 10.0 Hz), 6.05 (H-6, dd, J = 15.0, 10.0 Hz), 5.70 (H-7, dt, J = 15.0, 6.0 Hz), 2.05 (H2-8, m), 1.39 (H-9b, m), 1.53 (H-9a, m), 1.51 (H2-10, m), 3.71 (H-11, m), 1.13 (H3-12, d, J = 6.5 Hz), 3.67 (OCH3, s), 3.54 (2 x OCH3, s), 7.36-7.60 (10H, m); ESIMS m/z 673.48 [M - H]-.
 Compound 3c: pale, amorphous solid; 1 H NMR data (CD 3 OD): δ H 2.50 (H2-2, d, J = 6.5 Hz), 4.51 (H-3, m), 5.85 (H-4, dd, J = 15.0, 8.5 Hz) , 6.22 (H-5, dd, J = 15.0, 10.0 Hz), 6.05 (H-6, dd, J = 15.0, 10.0 Hz), 5.70 (H-7, dt, J = 15.0, 6.0 Hz), 2.05 (H2-8, m), 1.39 (H-9b, m), 1.53 (H-9a, m), 1.51 (H 2 -10, m), 3.71 (H-11, m), 1.13 (H 3- 12, d, J = 6.5 Hz), 3.67 (OCH 3 , s), 3.54 (2 × OCH 3 , s), 7.36-7.60 (10H, m); ESIMS m / z 673.48 [M−H] − .

화합물 4a의 제조. Preparation of Compound 4a .

화합물 4 (3.0 mg)를 2 mL 무수 MeOH에 녹였다. 화합물 4의 메틸 에스테르를 전술한 방법에 따라 제조하였다. 반응 혼합물을 진공 건조시키고, 60% MeOH를 함유한 H2O를 사용하여 ODS HPLC 상에서 정제하여 화합물 4의 메틸 에스테르를 얻었다 (화합물 4a, 2.3 mg).Compound 4 (3.0 mg) was dissolved in 2 mL anhydrous MeOH. The methyl ester of compound 4 was prepared according to the method described above. The reaction mixture was dried in vacuo and purified on ODS HPLC using H 2 O containing 60% MeOH to afford methyl ester of compound 4. (Compound 4a, 2.3 mg).

화합물 4a: 황색, 무정형 고체; 1H NMR 데이터 (CD3OD): δH 5.83 (H-2, d, J = 15.2 Hz), 7.26 (H-3, dd, J = 15.2, 10.3 Hz), 6.25 (H-4, dd, J = 15.2, 10.3 Hz), 6.21 (H-5, m), 2.20 (H2-6, m), 1.45 (H-7b, m), 1.56 (H-7a, m), 1.42 (H2-8, m), 3.71 (H-9, m), 1.14 (H3-10, d, J = 6.5 Hz), 3.70 (OCH3, s); ESIMS m/z 197.22 [M - H]-.
 Compound 4a: yellow, amorphous solid; 1 H NMR data (CD 3 OD): δ H 5.83 (H-2, d, J = 15.2 Hz), 7.26 (H-3, dd, J = 15.2, 10.3 Hz), 6.25 (H-4, dd, J = 15.2, 10.3 Hz), 6.21 (H-5, m), 2.20 (H 2 -6, m), 1.45 (H-7b, m), 1.56 (H-7a, m), 1.42 (H 2- 8, m), 3.71 (H -9, m), 1.14 (H 3 -10, d, J = 6.5 Hz), 3.70 (OCH 3, s); ESIMS m / z 197.22 [M−H] − .

(( SS )-) - MTPAMTPA 에스테르(화합물 4b) 제조.  Preparation of Ester (Compound 4b).

화합물 4a(1 mg)의 (S)-MTPA를 전술한 방법에 따라 (R)-(-)-MTPA-Cl로부터 제조하였다. 반응 혼합물을 진공 건조시키고, H2O에 재용해시키고, 및 EtOAc (3 x 3 mL)로 추출하였다. 추출물을 65% MeOH을 함유하는 H2O를 이동상으로 하여 ODS 분석 컬럼 상에서 정제하여 화합물 4b (0.8 mg)를 얻었다.( S ) -MTPA of Compound 4a (1 mg) was prepared from ( R )-(-)-MTPA-Cl according to the method described above. The reaction mixture was dried in vacuo, redissolved in H 2 O, and extracted with EtOAc (3 × 3 mL). The extract was purified on an ODS analytical column with H 2 O containing 65% MeOH as the mobile phase to give compound 4b. (0.8 mg) was obtained.

화합물 4b: 무정형 고체; 1H NMR 데이터 (CD3OD): δH 5.82 (H-2, d, J = 15.4 Hz), 7.23 (H-3, dd, J = 15.4, 10.7 Hz), 6.16 (H-4, dd, J = 15.4, 10.7 Hz), 6.06 (H-5, m), 2.11 (H2-6, m), 1.36 (H-7b, m), 1.56 (H-7a, m), 1.31 (H2-8, m), 5.13 (H-9, m), 1.33 (H3-10, d, J = 6.0 Hz), 3.71 (OCH3, s), 3.54 (OCH3, s), 7.39-7.57 (5H, m); ESIMS m/z 413.08 ([M - H]-), 414.95 [M + H]+.
 Compound 4b: amorphous solid; 1 H NMR data (CD 3 OD): δ H 5.82 (H-2, d, J = 15.4 Hz), 7.23 (H-3, dd, J = 15.4, 10.7 Hz), 6.16 (H-4, dd, J = 15.4, 10.7 Hz), 6.06 (H-5, m), 2.11 (H 2 -6, m), 1.36 (H-7b, m), 1.56 (H-7a, m), 1.31 (H 2- 8, m), 5.13 (H -9, m), 1.33 (H 3 -10, d, J = 6.0 Hz), 3.71 (OCH 3, s), 3.54 (OCH 3, s), 7.39-7.57 (5H m); ESIMS m / z 413.08 ([M−H] − ), 414.95 [M + H] + .

( R )- MTPA 에스테르(화합물 4c) 제조. (R) - MTPA ester (compound 4c) production.

화합물 4a(1 mg)의(R)-MTPA 에스테르를 전술한 방법에 따라 (S)-(+)-MTPA-Cl로부터 제조하였다. 화합물 4a의 (R)-MTPA 에스테르를 이동상으로 EtOAc/MeOH/n-헥산 (4:1:1)을 사용하는 반-분취 실리카 컬럼 상에서 정제하여 화합물 4c(0.77 mg)를 얻었다.( R ) -MTPA ester of compound 4a (1 mg) was prepared from ( S )-(+)-MTPA-Cl according to the method described above. The ( R ) -MTPA ester of compound 4a was purified on a semi-prepared silica column using EtOAc / MeOH / n -hexane (4: 1: 1) as the mobile phase to give compound 4c (0.77 mg).

화합물 4c: 무정형 고체; 1H NMR 데이터 (CD3OD): δH 5.84 (H-2, d, J = 15.3 Hz), 7.27 (H-3, dd, J = 15.3, 10.0 Hz), 6.23 (H-4, dd, J = 15.3, 10.0 Hz), 6.20 (H-5, m), 2.21 (H2-6, m), 1.46 (H-7b, m), 1.67 (H-7a, m), 1.37 (H2-8, m), 5.71 (H-9, m), 1.15 (H3-10, d, J = 6.0 Hz), 3.71 (OCH3, s), 3.54 (OCH3, s), 7.38-7.60 (5H, m); ESIMS m/z 413.07 [M - H]-, 414.93 [M + H]+. Compound 4c: amorphous solid; 1 H NMR data (CD 3 OD): δ H 5.84 (H-2, d, J = 15.3 Hz), 7.27 (H-3, dd, J = 15.3, 10.0 Hz), 6.23 (H-4, dd, J = 15.3, 10.0 Hz), 6.20 (H-5, m), 2.21 (H2-6, m), 1.46 (H-7b, m), 1.67 (H-7a, m), 1.37 (H 2 -8 , m), 5.71 (H- 9, m), 1.15 (H 3 -10, d, J = 6.0 Hz), 3.71 (OCH 3, s), 3.54 (OCH 3, s), 7.38-7.60 (5H, m); ESIMS m / z 413.07 [M−H] − , 414.93 [M + H] + .

한국생명공학연구원Korea Biotechnology Research Institute KCTC11975BPKCTC11975BP 2011061220110612

Claims (4)

하기 화학식 3으로 표시되는 지방산 화합물.
<화학식 3>
Figure 112013110654443-pat00009

상기 화학식 3에서 R1=R2 =R3=H 임.Fatty acid compound represented by the following formula (3).
(3)
Figure 112013110654443-pat00009

R 1 = R 2 in Chemical Formula 3 = R 3 = H 제 1항에 있어서 상기 화학식 3으로 표시되는 지방산 화합물이 B acillus subtilis, E. coli 및 Saccharomyces cerevisiae에 대하여 항균 활성을 가지는 것을 특징으로 하는 화학식 3으로 표시되는 지방산 화합물.According to claim 1, wherein the fatty acid compound represented by the formula (3) is B acillus subtilis , E. coli And Saccharomyces Fatty acid compound represented by the formula (3) characterized by having an antimicrobial activity against cerevisiae . 제 1항 또는 제 2항에 있어서 상기 화학식 3으로 표시되는 지방산 화합물이 기탁번호 KCTC 11975BP로 기탁된 해양 바실러스 속 미생물로부터 제조되는 것을 특징으로 하는 화학식 3으로 표시되는 지방산 화합물.The fatty acid compound represented by the formula (3) according to claim 1 or 2, wherein the fatty acid compound represented by the formula (3) is prepared from marine marine microorganisms deposited with the accession number KCTC 11975BP. 기탁번호 KCTC 11975BP로 기탁된 해양 바실러스 속 미생물을 0 .1% 효모 추출물, 0.1% 비프 추출물, 0.2% 트립톤, 1% 덱스트로즈, 염화나트륨 12 g/L, pH 7.6의 변형된 베넷 배지 에서 배양하는 단계; 및
상기 배양물을 원심분리한 상등액을 에틸아세테이트(EtOAc)로 추출한 후 에틸아세테이트 층을 증발 건조한 후 잔류 현탁물을 ODS open 칼럼크로마토그래피에 주입한 후, 이동상으로서 MeOH/H2O을 사용하여 단계적 분리한 후 semi-preparative ODS HPLC (50% MeOH 함유 H2O; 유속: 1.5 mL/min; 탐지기: RI)를 이용하여 분획하고 분획물을 13%MeOH-EtOAc; 5% MeOH-CHCl3 10% Me0H-CHCl3 6% MeOH-CHCl3를 이용한 isocratic 프로그램을 이용하여 순상 semi-preparative HPLC (유속: 1.3 mL/min; 탐지기: UV)로 정제하는 단계를 포함하는 것을 특징으로 하는 하기 화학식 3으로 표시되는 화합물의 지방산 화합물의 제조방법.
<화학식 3>
Figure 112013110654443-pat00010

상기 화학식 3에서 R1=R2 =R3=H 임.
Microorganisms of marine Bacillus sp. Deposited with accession number KCTC 11975BP were cultured in modified Bennett medium at 0.1% yeast extract, 0.1% beef extract, 0.2% tryptone, 1% dextrose, 12 g / L sodium chloride, pH 7.6 Making; And
The culture supernatant of the culture was extracted with ethyl acetate (EtOAc), the ethyl acetate layer was evaporated to dryness, the residue was injected into ODS open column chromatography, and phase separation was performed using MeOH / H 2 O as a mobile phase. Fractions were then purified using semi-preparative ODS HPLC (H 2 O with 50% MeOH; flow rate: 1.5 mL / min; detector: RI) and the fractions were 13% MeOH-EtOAc; Purifying by normal phase semi-preparative HPLC (flow rate: 1.3 mL / min; detector: UV) using an isocratic program with 5% MeOH-CHCl 3 10% Me0H-CHCl 3 6% MeOH-CHCl 3 A method for producing a fatty acid compound of a compound represented by the following formula (3).
(3)
Figure 112013110654443-pat00010

R 1 = R 2 in Chemical Formula 3 = R 3 = H
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Publication number Priority date Publication date Assignee Title
WO2006070891A1 (en) 2004-12-28 2006-07-06 Meiji Seika Kaisha, Ltd. Novel strain conferring anti-disease properties to host and bacterial cell composition

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Bioorganic and Medicinal Chemistry Letters, Vol.21, pp.3382-3385(Epub.2010.12.16.) *
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