Human risk assessment using lipid adduct : Lipid adduct를 利用한 人體 危害性 評價

송현수 成均館大學校 大學院 2002 국내박사

현대사회의 산업이 발달할수록, 인체는 환경 중의 다양한 발암물질에 계속적으로 노출되어, 이에 따른 발암 위해성 역시 증가하고 있는 추세이다. 그러므로 보다 정확하고 편리한 인체의 발암 위해성 평가 방법이 중요한 문제로서 부각되고 있다. 본 논문에서는 새로운 표징물질로서 lipid adduct와 TG adduct를 이용한 새로운 발암 위해성 평가 방법을 제시하였다. 기존의 정량법 중, 경제적이고 편리하며 정확한 ELISA를 이용하여 BPDE-lipid in vitro와 BPDE-TG in vitro를 정량 해내었다. 그러나 이러한 측정은 BPDE-lipid in vivo에서는 전혀 감도가 나타나지 않았으며, 단지 BPDE-TG in vivo에서만 감도를 보였다. ELISA에서의 BPDE 정량의 감도를 증가시키기 위하여, [3H] BaP투여 마우스로부터 얻어진 BPDE-TG를 이용하여 여러 가지 조건을 시도하였으며, 이러한 시도로부터 가장 최적의 조건을 확립하였다. 이렇게 확립된 조건을 바탕으로, 인체혈액과 조직에서 BPDE를 측정함으로써 인체 위해성평가를 수행하였다. 그 결과, 흡연자와 암 조직에서의 BPDE 수치가 비흡연자와 정상조직에 비해서 높게 나타났다. 그러나, 본 연구에서의 위해성 평가는 몇 가지 해결해야 할 문제점이 제시되었으며, 차후에 이런 문제점을 해결하여 보완한다면 인체 위해성 평가에서의 보다 정확한 응용이 가능하리라 예상된다. 비록 인체에 적용하는 점에서는 현재 몇 가지 문제점을 가지고 있지만, lipid나 TG adduct를 ELISA에 적용하기 위한 조건을 설립하고, 이로부터 동물에서의 노출 정도를 측정하였다는 점, 그리고 lipid와 TG가 발암과정에 연관이 된다는 가능성을 증명하였다는 점에서 본 연구는 새로운 시도라 할 수 있다. 또한 최초의 lipid adduct와 TG adduct를 이용한 위해성 평가라는 점에서 본 연구는 매우 의미있는 연구라 사료된다. As humans are continuously exposed to a variety of carcinogens and are in a high risk for cancer, it was recognized the importance of cancer risk assessment. To develop an accurate and convenient risk assessment method, BPDE-lipid and BPDE-TG as new biomarker was applied by ELISA. BPDE was quantitated in BPDE-lipid and BPDE-TG adduct from animals treated with [3H] BaP and human exposed to BaP environmentally by ELISA. In the case of in vitro, BPDE-lipid and BPDE-TG were operated in ELISA, properly. But, despite trial of various conditions, BPDE-lipid in vivo was not reacting in ELISA. In BPDE-TG in vivo, to increase the sensitivity of the assay, various conditions were tested and established. Through this established conditions, BPDE was quantitated by ELISA in human blood and tissue. Here, this result indicates that BPDE-TG can use as new biomarker. And BPDE-TG is measured by ELISA in animal and human. Although cancer risk assessment using BPDE-TG by ELISA has a few point, this developed method were more convenient than the existing method. These issues also include that lipid and TG were related with carcinogenesis. It was suggested that the more accurate and convincing methodology for cancer risk assessment was developed and applied to estimate the human exposure to carcinogens and risk assessment in this study.

Baylis-Hillman Adducts로부터 Cinnamyl Alcohols과 Dihydroquinolines의 합성에 관한 연구

김형식 전남대학교 대학원 2001 국내석사

Baylis-Hillman adduct와 trifluoroacetic acid를 30-70°C에서 반응 시키면 적당한 수율로 rearrane된 cinnamyl alcohol이 입체 선택적으로 얻어진다. Ester group을 가지고 있는 유도체들로부터는 E-form의 allylic alcohol을 51-72% 수율로 얻었고, nitrile을 가지고 있는 경우에는 Z-form으로 27-40%의 다소 감소된 수율로 얻을 수 있었다. ortho-Halobenzaldehyde의 Baylis-Hillman adduct로부터 얻어진 아세틸화된 Baylis-Hillman adduct와 여러가지 amine 유도체와의 반응으로부터는 처음에 1,2-dihydroquinoline을 얻을 수 있었다. 그러나, 이 화합물은 서서히 보다 안정한 1,4 -dihydroquinoline으로 isomerize되었다. The reaction of the Baylis-Hillman adducts and trifluoroacetic acid at 30-70 ℃ gave the rearranged cinnamyl alcohols stereoselectively in moderate yields. Ester derivatives gave E-form allylic alcohols stereoselectively in 51-72% isolated yields while nitrile-containing Baylis-Hillamn addcuts afforded Z-form analogues in somewhat diminished yields (27-40%). The reaction of the acetates of the Baylis-Hillman adducts of ortho-halobenzaldehydes and some amines gave the N-substituted 1,2-dihydroquinolines as the initial products. However, these compounds slowly isomerises to the more stable 1,4-dihydro analogues. In some case the 1,2-dihydroquinolines could be transformed to the corresponding quinolines depending on the N-substituent.

Identification and characterization of DNA adducts of 1-bromopropane in vivo and in vitro

Nepal, Mahesh Raj Yeungnam University 2016 국내석사

Baylis-Hillman adducts 및 aza-Baylis-Hillman adducts의 Heck 반응

김정미 전남대학교 대학원 2009 국내석사

본 논문은 Baylis-Hillman adducts의 화학적 변환 연구를 통한 β-아릴 치환된 Baylis-Hillman adducts의 합성과 β-아릴 치환된 aza-Baylis-Hillman adducts의 합성, 그리고 다양한 생리활성이 기대되는 헤테로고리 화합물들, 특히 다양한 치환체가 위치 선택적으로 도입된 pyrrole 및 pyrimidine 유도체의 합성방법 개발에 대해 다루고 있다. 우리는 β-아릴 치환된 Baylis-Hillman adducts의 합성을 위하여 Baylis-Hillman adducts와 aryl halides과의 Pd을 이용한 분자 간 Heck 반응을 이용하였다. 여러 연구진의 시도에도 불구하고 분자 간 Heck 반응을 통한 β-아릴 치환된 Baylis-Hillman adducts의 합성은 성공한 사례가 없었으나 우리는 Pd(OAc)₂ (15 mol%) / TBAB (1.0 equiv.) / KOAc (3.0 equiv.) / CH₃CN / reflux 조건하에서 Baylis-Hillman adducts와 aryl iodides의 분자 간 Heck 반응을 수행하여 β-아릴 치환된 Baylis-Hillman adducts를 합성할 수 있었다. 우리는 Baylis-Hillman adducts의 Heck 반응의 긍정정인 결과를 좀 더 다양한 substrate에 적용해보고자 aza-Baylis-Hillman adducts의 분자 간 Heck 반응을 시도하였고 Pd(OAc)₂ (5 mol%)/ PPh₃ (0.1 equiv.)/ TBAB (1.0 equiv.)/ KOAc (3.0 equiv.)/ CH₃CN/ reflux 조건하에서 aza-Baylis-Hillman adducts와 aryl iodides의 분자 간 Heck 반응을 성공적으로 수행할 수 있었고 이를 통해 β-아릴 치환된 aza-Baylis-Hillman adducts를 합성할 수 있었다. 우리는 Baylis-Hillman adducts에서 유도된 α-methylene-γ-keto ester 화합물과 amine 유도체와의 반응을 통하여 위치 선택적으로 다양한 치환체를 도입한 pyrrole 유도체를 아래와 같은 과정을 통하여 합성할 수 있었다. (ⅰ) Baylis-Hillman adducts의 bromination을 하여 cinnamyl bromide 유도체를 얻은 후, (ⅱ) cinnamyl bromide 유도체와 bezaldehyde의 In-mediated Barbier reaction한 후, 이를 (ⅲ) PCC oxidation하여 α-methylene-γ-keto ester 화합물의 합성, (ⅳ) 합성된 α-methylene-γ-keto ester와 amine 유도체가 AcOH / toluene / 80-90 oC 조건하에서 반응하여 enamine 중간체 형성, Michael type cyclization, aerobic oxidation을 거쳐 최종 생성물인 다중 치환된 pyrrole 유도체를 얻을 수 있다. 우리는 Baylis-Hillman acetate 와 amidine 유도체를 이용하여 K₂CO₃ / tert-butanol / reflux 조건에서 2,4,5-삼중 치환된 pyrimidine 유도체를 좋은 수율로 합성할 수 있었다. 우리는 Baylis-Hillman acetate 로부터 합성된 alkylidene uracil 유도체를 PCC (2.0 equiv.)/ CH₂Cl₂/ reflux 조건에서 allylic oxidation 시켜 5-aroyl uracil 유도체 합성할 수 있었다. 출발물질로 사용된 alkylidene uracil은 Baylis-Hillman acetate의 1차 위치에 benzyl amine이나 aniline을 도입한 후, KOCN, PhCNO 등을 도입하여 urea 유도체를 합성, 이를 K₂CO₃ (0.2 equiv.)/ DMF/ r.t. 조건에서 condensation하여 합성하였다. 우리는 Baylis-Hillman adducts를 N,N-dimethylacetamide-dimethyl-acetal (DMA-DMA) (2 equiv.)/ toluene / 80-90 oC 조건에서 Eschenmoser-Claisen rearrangement시켜 γ,δ-unsaturated amide 합성 할 수 있었다. β-Aryl-substituted Baylis-Hillman adducts were synthesized via the Heck type reaction of Baylis-Hilman adducts and aryl iodides under the influence of Pd(OAc)₂ (15 mol%)/ TBAB (1.0 equiv.)/ KOAc (3.0 equiv.) in refluxing CH₃CN in moderate yields as cleanly separable E/Z mixture. This is the first successful intermolecular Heck reaction of Baylis-Hillman adducts. We also prepared some β-aryl N-tosyl aza-Baylis-Hillman adducts via the Heck type reaction of aza-Baylis-Hillman adducts and aryl iodides under the influence of Pd(OAc)₂ (5 mol%)/ PPhc₃(0.1 equiv.)/ TBAB (1.0 equiv.)/ KOAc (3.0 equiv.) in refluxing CH₃CN in moderate yields as E/Z mixture. Poly-substituted pyrrole derivatives were synthesized from Baylis-Hillman adducts via the consecutive reactions comprised of (ⅰ) bromination of Baylis-Hillman adducts to cinnamyl bromide, (ⅱ) In-mediated Barbier reaction with aldehyde, (ⅲ) PCC oxidation to α-methylene-γ-keto ester, (ⅳ) reaction with amine to form enamine intermediated, (ⅴ) Michael type cyclization, (ⅵ) and the final concomitant aerobic oxidation. Synthesis of 2,4,5-trisubstituted pyrimidines was carried out from the reaction of Baylis-Hillman acetates and amidine derivatives in a one-pot reaction in moderate yields. Various biologically interesting 5-arylidene- and 5-aroyluracil derivatives were also synthesized starting from Baylis-Hillman adducts. The Synthesis was carried out as follows: (ⅰ) introduction of amine derivatives at the primary position of Baylis-Hillman adducts, (ⅱ) conversion into their urea and thiourea derivatives, (ⅲ) cyclization to 5-arylideneuracil and thiouracil derivatives, and (ⅳ) PCC oxidation to the corresponding 5-aroyluracil and 5-aroyl-2-thiouracil. Eschenmoser-Claisen rearrangement of Baylis-Hillman adducts was examined. Various 2-benzylidene 1,5-dicarbonyl compounds and 2-methylene 1,5-dicarbonyl derivatives were synthesized in moderate yields from the reaction of Baylis-Hillman adducts and cinnamyl alcohol derivatives with N,N-dimethylacetamide-dimethylacetal (DMA-DMA). In summary, the first intermolecular Heck type arylation of Baylis-Hillman and aza-Baylis-Hillman adducts has been carried out successfully and we opened an easy access to β-aryl-substituted Baylis-Hillman adducts. In addition, expedient synthetic method for the preparation of poly-substituted pyrroles, 2,4,5-trisubstituted pyrimidines, and various 5-aroyluracil derivatives were developed.

Baylis-Hillman adducts 로부터 헤테로고리 화합물의 합성에 관한 연구

이가영 전남대학교 대학원 2002 국내석사

ortho-Position에 nitro group을 갖고 있는 benzaldehydes와 ethyl acrylate로부터 유도된 Baylis-Hillman adducts를 trifluaroacetic acid 용매에서, 60-70℃ 정도로 가열하여 3-ethoxycarbonyl-4-hydroxyquinoline N-oxides를 좋은 수율로 합성할 수 있었다. 같은 출발물질을 이용하여 EtOH, 350nm의 photoreaction으로 3-ethoxycarbonyl-4-hydroxyquinolines을 합성할 수 있었다. o-Nitrobenzaldehydes와 methyl vinyl ketone, phenyl vinyl sulfone등을 이용하여 만들어진 Baylis-Hillman adducts로부터 trifluoroacetic acid, triflic acid(cat), 30-40℃ 조건에서 좋은 수율로 3-substituted-4-hydroxy quinoline N-oxides를 합성하였고 그에 대한 mechanism은 N-hydroxyisoxazo line을 거치는 과정이라 제시할 수 있었다. o-Nitrobenzaldehydes로부터 얻어진 Baylis-Hillman adducts를 이용하여 Zn와 AcOH, TFA(cat) 조건에서 좋은 수율로 3-(acetoxymethyl)-(1H)-quinol-2-ones를 합성할 수 있었다. 반응 메카니즘은 다음과 같이 제시할 수 있다. Nitro 작용기가 amino 작용기로 환원되고, 산촉매에 의해 amide bond formation이 일어난 후, acetic acid가 conjugate addition 되고 dehydration이 일어나 3-(alkoxymethyl)-(1H)-quinol-2-ones가 얻어지는 것으로 생각된다. 이와 유사하게 o-nitrobenzaldehydes로부터 얻어진 Baylis-Hillman adducts를 이용하여 SnCl₂와 ROH 조건에서 3-(alkoxymethyl)-(1H)-quinol-2-ones을 합성할 수 있었다. o-Nitrobenzaldehydes로부터 얻어진 Baylis-Hillman adducts를 이용하여 Zn/NH₄Cl 조건에서 3-ethoxycarbonylquinoline N-oxides를 합성할 수 있었다. 반응 메카니즘은 다음과 같이 제시할 수 있다. Nitro 작용기가 nitroso 작용기로 환원되고 Michael addition이 일어난다. 그 반응물은 dehydration과 deprotonation을 거쳐 3-ethoxycarbonylquinoline N-oxides가 얻어지는 것으로 생각된다. 다양한 aldehyde 유도체와 2-cyclohexen-1-one으로부터 유도된 Baylis-Hillman adducts를 acetylation하여 K₂CO₃와 DMF 조건에서 좋은 수율로 2-benzylphenols를 합성할 수 있었다. 반응 메카니즘은 다음과 같이 제시할 수 있다. 아세트산 분자가 제거되고 1,5-proton transfer가 일어난 후 keto-enol toutomerization이 되어 2-benzylphenols이 합성되어 진다. 2-(2-halobenzyl)-phenols로부터 intramolecular nucleophilic aromatic substitution 반응을 이용하여 xanthene 유도체들을 얻을 수 있었다. Salicylaldehyde 유도체들과 2-cyclohexen-1-one으로부터 DMAP(cat)/aqueous THF조건에서 2, 3, 4, 4a-tetrahydroxanthen-1-one 유도체를 합성할 수 있었다. 반응 메카니즘은 다음과 같이 제시할 수 있다. Salicylaldehydes와 2-cyclohexen-1-one으로부터 Baylis-Hillman adducts가 유도되고 ortho의 hydroxy group이 Michael addition을 한 후 dehydration 되어 2, 3, 4, 4a-tetra hydroxanthen-1-ones가 얻어진다. The reaction of the Baylis-Hillman adducts of 2-nitrobenz aldehydes and trifluoroacetic acid at 60-70℃ gave 3-ethoxycarbonyl-4-hydroxyquinoline N-oxide derivatives in good to moderate yields. The same adducts under photo-irradiation conditions gave the quinoline derivatives as the major product instead of the corresponding N-oxides. Synthesis of 3-substituted-4-hydroxyquinoline N-oxides was carried out in a similar reaction conditions. We used Baylis-Hillman adducts derived from 2-nitrobenzaldehydes and some activated vinyls such as methyl vinyl ketone and phenyl vinyl sulfone. In the reaction trifluoro methanesulfonic acid must be used as a catalyst. The Baylis-Hillman adducts of 2-nitrobenzaldehydes and Zn/AcOH gave 2(1H)-quinolinone derivatives in good yields. The reaction might proceed via the successive reaction : (1) reduction of the nitro functional group to amino fuctionality, (2) amide-bond formation, (3) conjugate addition of acetic acid, and finally (4) dehydration. Similarly, reduction of the Baylis-Hillman adducts of 2-nitrobenzaldehydes with SnCl₂/ ROH system afforded the corresponding 3-alkoxymethyl 2(1H)-qinolinones in good yields. The Baylis-Hillman adducts of 2-nitrobenzaldehydes and Zn / NH₄Cl / aqueous THF gave 3-ethoxycarbonylquinoline N-oxide in moderate yields. The reaction might proceed via the successive reaction : (1) reduction of the nitro functional group to nitroso functionality, (2) Michael type addition, (3) dehydration, and finally (4) deprotonation. The Baylis-Hillman acetates derived from 2-cyclohexen-1-one and arylaldehydes were converted into 2-benzylphenols in the presence of K₂CO₃ in DMF. The reaction might proceed via the following sequence : (1) elimination of acetic acid, (2) 1,5-proton transfer, and finally (3) keto-enol toutomerization. The reaction of 2-cyclohexen-1-one and salicylaldehydes in the presence of DMAP in aqueous THF affords the corresponding 2,3,4,4a-tetrahydroxanthen-1-ones in moderate yields. The reaction might proceed via the following sequence : (1) Baylis-Hillman reaction of 2-cyclohexen-1-one with salicylaldehydes, (2) Michael type addition of the phenolic oxygen, and finally (3) dehydration. As a conclusion in this thesis synthesis of various heterocyclic compounds was achieved from the Baylis-Hillman adducts.

베일리스-힐만 부가생성물로부터 α-pyrone 유도체의 합성

김성진 전남대학교 대학원 2008 국내석사

이 논문에서는 Baylis-Hillman adducts를 출발물질로 하여 생리학적으로 중요한 α-pyrone 유도체를 합성 하였다. 또한, 다중 치환된 pyrrole 유도체와 phenol 유도체의 효과적인 합성 방법도 개발하였다. Baylis-Hillman adducts로부터 3,5,6-trisubstituted α-pyrone의 합성은 Baylis-Hillman adducts의 primary 위치에 다양한 케톤 화합물을 도입 하고, TFAA를 이용한 lactonization, 그리고 PCC (pyridinium chlorochlomate)를 이용한 allylic oxidation 반응으로 이루어졌다. MVK에서 유도된 Baylis-Hillman acetates와 diethyl iminodiacetate를 K2CO3 조건에서 반응시켜 SN2' 반응이 일어난 화합물을 합성하고, 연이은 분자내 aldol type 반응으로 pyrrolidine 유도체의 diastereomeric mixture를 합성할 수 있었다. 이렇게 합성된 pyrrolidine 유도체는 산 조건에서 탈수반응과 double bond isomerization 과정을 경위하여 1,2,3,4-tetrasubstituted pyrrole 유도체로 변화되었다. Baylis-Hillman adducts를 Friedel-Crafts 반응으로 benzene을 primary 위치에 도입하여 trisubstituted alkene 유도체를 합성하였다. 이렇게 합성된 trisubstituted alkene 유도체를 NBS와 AIBN으로 allylic bromination 시킨 후에 nucleophie을 도입하여 β-phenyl Baylis-Hillman adducts를 합성할 수 있었다. MVK에서 유도된 Baylis-Hillman acetates의 primary 위치에 K2CO3, DMF, heating 조건에서 다양한 케톤 화합물을 도입 하고, 연이은 분자내 aldol type 반응과 탈수 반응, 그리고 double bond isomerization 과정을 통하여 다중 치환된 phenol 유도체를 합성할 수 있었다. 요약하자면, 우리는 Baylis-Hillman adducts의 primary 위치에 적당한 nucleophiles (ketone과 iminodiacetate)을 연속적으로 도입하고 중간체의 다음 조작으로 Baylis-Hillman adducts로부터 다양한 α-pyrone과 다중 치환된 pyrrole 유도체를 합성할 수 있었다. 또한, Friedel-Crafts 반응과 allylic bromination, 친핵성 치환반응을 이용하여 β-치환된 Baylis-Hillman adducts를 손쉽게 합성할 수 있었다. 마지막으로 Baylis-Hillman adducts와 케톤 화합물의 반응을 통하여 다양한 phenol 유도체를 합성할 수 있었다. Synthesis of biologically important α-pyrone derivatives starting from the Baylis-Hillman adducts was examined in this thesis. In addition, an efficient synthetic procedure for poly-substituted pyrroles and phenols was also developed. 3,5,6-Trisubstituted α-pyrones were synthesized from the Baylis- Hillman adducts. The synthesis was carried out via the sequential introduction of various ketones at the primary position of Baylis-Hillman adducts, lactonization, and the following oxidation with PCC (pyridinium chlorochlomate). The reaction of Baylis-Hillman acetates and diethyl iminodiacetate in the presence of K2CO3 produced the corresponding diastereomeric mixtures of pyrrolidines via the successive SN2' reaction and intramolecular aldol type reaction. The pyrrolidines could be converted into 1,2,3,4-tetrasubstituted pyrrole derivatives under acidic conditions in moderate yields. We developed an efficient synthetic methodology of β-phenyl -substituted Baylis-Hillman and aza-Baylis-Hillman adducts. The synthesis of these compounds was carried out via the sequential reaction of Friedel-Crafts reaction, allylic bromination, and the final nucleophilic substitution reaction with water or amine nucleophiles. We also developed a facile and regioselective synthetic procedure for poly-substituted phenol derivatives starting from the Baylis-Hillman adducts via the formal [4+2] annulation protocol as the key step. The synthesis was performed according to the following sequence: (i) introduction of diaryl or monoaryl ketones at the primary position of Baylis-Hillman adducts, (ii) intramolecular aldol condensation, and finally (iii) keto-enol tautomerization, and isomerization process. In summary, we synthesized various α-pyrones and poly- substituted pyrrole and phenol derivatives from Baylis-Hillman adducts via the sequential introduction of suitable nucleophiles (ketone and iminodiacetate) at the primary position of Baylis-Hillman adducts and the following manipulation of the intermediates.

Baylis-Hillman Adducts와 다양한 친핵체들의 반응에 관한 연구

이홍정 전남대학교 대학원 2001 국내박사

다양한 benzaldehyde들의 N-tosylimine 유도체로부터 Baylis-Hillman adducts 를 합성하여, 황산 촉매하에서 방향족 화합물과 Friedel-Crafts 반응을 시켜서 입체 선택적으로 2-benzyl substituted olefins을 비교적 좋은 수율로 얻을 수 있었다. 생성되어진 trisubstituted olefin유도체들의 stereochemistry는 출발물질로 사용된 Baylis-Hillman adducts의 electron withdrawing group(EWG)에 따라 뚜렷이 구분된다. EWG이 ethoxycarbonyl인 경우 E- form이 우선적으로 얻어지며, 반면에 nitrile group인 경우 Z- form이 선택적으로 얻어진다. N-tosylimines의 Baylis-Hillman adducts와 N,N-dimethylformamide dimethylacetal (DMF-DMA)과의 반응으로 N-methyl-N-tosyl allylic amine유도체를 입체 선택적으로 얻을 수 있었다. 반응 메카니즘은 잠정적으로 다음과 같이 제시할 수 있다. (1) N,N-dimethylformamide dimethylacetal에 소량 존재하는 methoxide 이온이 Michael type addition으로 반응하여 tetrahedral intermediate를 만들고, (2) tosylamide anion이 제거된다. (3) 이때 생성된 tosylamide anion이 출발물질인 Baylis-Hillman adducts에 S_N2’ type으로 addition되고, 이와 동시에 tosylamide anion이 regeneration된다. 이렇게 만들어진 화합물에 최종적으로 N-methylation이 일어나서 N-methyl-N-tosylcinnamyl amine유도체들이 얻어진다. Benzaldehyde으로 부터 합성된 Baylis-Hillman adducts의 N,N-dimethyl formamide dimethylacetal과의 반응으로부터 입체선택적으로 N,N-dimethylcinnamylamme유도체들을 비교적 좋은 수율로 얻었다. 반응 메카니즘은 잠정적으로 다음과 같이 제시 할 수 있다. N,N-Dimethylformamide dimethylacetal의 methoxy group이 Baylis-Hillman adducts의 alcohol작용기와 exchange되어 mixed acetal intermediate를 형성한다. 형성된 중간체의 dimethylamino group이 concerted Michael type addition되고 동시에 methyl formate가 제거됨으로써 N,N-dimethylcinnamyl amine유도체들을 만든다. ortho-Halobenzaldehyde의 N-tosylimine으로부터 유도된 Baylis-Hillman adducts를 N,N-dimethylformamide에서 K₂CO₃와 TsNH₂와의 반응으로 quinoline유도체들을 좋은 수율로 얻었다. 이 반응은 initial products로 rearranged N-tosylcinnamyl amine유도체들이 형성되고 intramolecular nucleophilic aromatic substitution reaction으로 1,2-dihydroquinoline유도체를 만든다. 마지막 단계로 p-toluenesulfinic acid가 제거됨으로써 해당하는 quinoline유도체가 합성된다. Friedel-Crafts reaction of aromatic compounds with the Baylis-Hillman adducts of N-tosylimine derivatives in the presence of sulfuric acid provided a stereoselective methodology for the preparation of 2-benzyl substituted olefins in moderate yields. The stereochemistry of the produced trisubstituted olefins differs markedly according to the electron withdrawing group of the starting Baylis-Hillman adducts. Where the EWG is ethoxycarbonyl group, E form olefin was obtained exclusively, while the corresponding Z form was isolated in the case of nitrile derivative. The reaction of N,N-dimethylformamide dimethylacetal and the Baylis-Hillman adducts of N-tosylimines afforded N-methyl-N-tosyl allylic amine derivatives stereoselectively in moderate yields. The reaction mechanism could be proposed as follows. Michael type addition methoxide, which might be present in N,N-dimethylformamide dimethylacetal in small amounts to give a tetrahedral intermediate, elimination of the tosylamide anion, S_N2' type addition of the tosylamide to the starting Baylis-Hillman adducts and concomitant regeneration of the tosylamide anion, and finally N-methylation of the resulting N-tosylcinnamylamine derivatives. The reaction of the Baylis-Hillman adducts of benzaldehydes and N,N-dimethylformamide dimethylacetal gave the corresponding N,N-dimethylcinnamyl amine derivatives in good to moderate yields stereoselectively. The reaction mechanism could be proposed as follows. Exchange of methoxy group in N,N-dimethylformamide dimethylacetal with the Baylis-Hillman adduct to form the corresponding mixed acetal intermediate, concerted Michael type addition of dimethylamino group and concomitant elimination of methyl formate gave the products. The reaction of the Baylis-Hillman adducts of ortho-halobenzaldehyde N-tosylimines in the presence of potassium carbonate and tosylamide in N,N-dimethylformamide gave the corresponding quinoline derivatives in good yields. The reaction gave the rearranged N-tosylcinnamyl amine derivatives as the initial products, which undergo subsequent intramolecular nucleophilic aromatic substitution reaction to generate the corresponding 1,2-dihydroquinoline analogues. The final step is the elimination of p-toluenesulfinic acid.

Baylis-Hillman Adducts로부터 나프탈렌과 o-히드록시아세토페논의 합성

임양진 전남대학교 대학원 2003 국내석사

Baylis-Hillman acetates와 primary nitroalkanes을 K₂CO₃, DMF 조건하에서 연속적인 S_N2'-S_NAr-elimination 단계를 거쳐서 2-substituted naphthalenes을 합성하였다. Baylis-Hillman acetates와 sulfonyl group을 가지고 있는 active methylene compound로부터 연속적인 (1) S_N2', (2) S_NAr 반응, (3) p-toluenesulfinic acid의 elimination 단계를 거쳐서 naphthalenes 유도체를 합성하였다. Baylis-Hillman acetates로부터 연속적인: (1) diethyl malonate 또는 ethyl nitroacetate의 S_N2'type반응; (2) manganese(III) acetate을 이용한 radical cyclization, (3) Nal/O₂조건에서의 방향족화 또는 nitrous acid elimination 단계를 거쳐서 1,3-Disubstituted naphthalene 유도체를 쉽게 합성하였다. Baylis-Hillman acetates에 DBU나 DBN이 특별한 반응성을 보여주었다. 이 반응으로부터 ε-caprolactam 유도체와 γ-butyrolactam 유도체가 각각 얻어짐을 알 수 있었다. Baylis-Hillman acetates로부터 ortho-hydroxyacetophenones 유도체를 합성하였다. methyl vinyl ketone에서 얻어진 Baylis-Hillman acetates에 2,4-pentanedione을 반응시켜 S_N2' 생성물을 합성한 후, 이 물질로부터 K₂CO₃, DMF조건하에서 ortho-hydroxyacetophenone 유도체를 합성하였다. Baylis-Hillman acetates와 β-diketones 혹은 diethyl malonate을 반응시켜 S_N2'type 생성물을 합성한 후, 이 물질로부터 LiHMDS, THF 조건하에서 4-arylidenecyclohexanone-1,3-dione을 합성하였다. 결론적으로 본 논문에서는, Baylis-Hillman adduct로부터 naphthalenes, ortho-hydroxyacetophenones, cyclohexane-1,3-diones 유도체를 합성하였다. The reaction of the Baylis-Hillman acetates and primary nitroalkanes in the presence of potassium carbonate in N,N-dimethylform -amide afforded 2-substituted naphthalenes in good yields via the successive S_N2'-S_NAr-elimination strategy. Naphthalene derivatives were synthesized from the Baylis-Hillman acetates and sulfonyl group-containing active methylene compound. Naphthalenes are formed via the successive three step mechanism: (1) S_N2' reaction of sulfonyl group-containing active methylene compounds, (2) S_NAr reaction, and (3) elimination of p-toluenesulfinic acid. 1,3-Disubstituted naphthalene derivatives could be easily synthesized from Baylis-Hillman acetates by successive reaction: (1) S_N2' type reaction with diethyl malonate or ethyl nitroacetate; (2) manganese(III) acetate-assisted radical cyclization and (3) aromatization with NaI/O₂system or elimination of nitrous acid. DBU and DBN showed unusual reactivity toward Baylis-Hillman acetates. From the reaction ε-carprolactam and γ- butyrolactam derivatives were formed, respectively. Synthesis of ortho-hydroxyacetophenone derivatives from the Baylis-Hillman acetates was carried out. The reaction of the Baylis-Hillman acetates derived from methyl vinyl ketone and 2,4-pentanedione afforded the corresponding S_N2' products. The compounds could be transformed into ortho-hydroxyacetophenones in the presence of K₂CO₃in N,N-dimethylform -amide. The reaction of the Baylis-Hillman acetates and some β-diketones, β-ketoesters, or diethyl malonate afforded the corresponding S_N2' type substitution products. These compounds could be transformed into 4-arylidenecyclohexanone-l,3-diones in moderate yields. As a conclusion in this thesis synthesis of naphthalenes, ortho-hydroxyacetophenones, and cyclohexane-1,3-diones was carried out from the appropriate Baylis-Hillman adducts.

모리타-베일리스-힐만 부가생성물로부터 다양한 헤테로고리 화합물의 합성

이현승 전남대학교 대학원 2012 국내박사

The Baylis-Hillman reaction was found by the German chemists Baylis and Hillman in 1972. The Baylis-Hillman reaction is an effective carbon-carbon bond-forming reaction between the alpha-position of activated vinyl compounds and some electrophiles such as aldehydes and N-tosylimines. The alpha-position of the activated vinyl compounds is intrinsically nucleophilic although the nucleophilic nature is very weak. However, the nucleophilicity of the alpha-position of activated vinyl compounds can be increased dramatically by in-situ formation of the corresponding zwitterionic species between the vinyl compounds and DABCO. A few years before the finding of Baylis and Hillman, a Japanese scientist Morita used phosphine instead of DABCO, and the Baylis-Hillman reaction is nowadays called as Morita-Baylis-Hillman reaction most frequently. The Morita-Baylis-Hillman reaction did not receive much attention at the earliest stage presumably due to some limitations of the original reaction including a slow reaction rate, limitations in electrophilic components, and etc. In the early 1990th Basavaiah and Kim groups started independently deep investigations on the Morita-Baylis-Hillman reaction itself and chemical transformations using the Morita-Baylis-Hillman adducts. The two research groups shed much light to the Morita-Baylis-Hillman chemistry by publishing many brilliant research results on the chemical transformations of the Morita-Baylis-Hillman adducts. In this Ph. D. thesis, many useful and novel chemical transformations with the Morita-Baylis-Hillman adducts and related compounds are described. In Chapter 1, a brief introduction of Morita-Baylis-Hillman reaction and a palladium-catalyzed reaction are provided. The Chapter 2 dealt with the results on the synthesis of various benzazepine derivatives starting from the suitably-modified Morita-Baylis-Hillman adducts. Initially, we examined the palladium(0)-catalyzed synthesis of 7H-benzo[3,4]azepino[1,2-a]indole-6-carboxylic acid derivatives from indole moiety-containing Morita-Baylis-Hillman adducts. Poly-fused heterocyclic compounds bearing an indole moiety were known to possess many interesting biological activities including hepatitis C virus (HCV) NS5B polymerase inhibitory activity. The required starting materials were prepared by the introduction of various indole derivatives at the primary position of the Morita-Baylis-Hillman acetate of 2-bromobenzaldehyde in good yields in a straightforward manner. A subsequent palladium(0)-catalyzed Heck-type reaction was conducted efficiently under the influence of Pd(OAc)2/TBAB in the presence of K2CO3 in DMF. Seven-membered ring was formed when the 2-position of the indole moiety is un-substituted while an eight-membered ring was formed when we use 2-methylindole derivative. The scope of the reaction was also investigated deeply by replacing the indole moiety of the above starting materials with isatins, benzimidazoles, and imidazoles. Various types of poly-fused heterocyclic compounds were synthesized from the modified Morita-Baylis-Hillman adducts with isatins, benzimidazoles, and imidazoles. Overall, we found that a palladium(0)-catalyzed coupling reaction of suitably-modified Morita-Baylis-Hillman adducts could provide an easy access toward many interesting seven- and eight-membered ring-containing poly-fused heterocyclic compounds. In addition, various tetracyclic oxindole derivatives were synthesized by reduction of the poly-fused isatin derivatives with NaBH4. During the reduction of isatin moiety with NaBH4 we found the fact that a facile aerobic oxidation of oxindole to isatin can occur under the same reduction conditions. In Chapter 3, a palladium(0)-catalyzed synthesis of benzo[c]pyrimido[1,6-a]azepine scaffold from the Morita-Baylis-Hillman adducts bearing an uracil moiety at the primary-position is described. The requisite starting materials were prepared by the introduction of various uracil derivatives at the primary position of the Morita-Baylis-Hillman acetate of 2-bromobenzaldehyde and a subsequent N-benzylation. The palladium(0)-catalyzed coupling reaction involved a novel and hitherto unknown intramolecular 6-arylation of uracil moiety. In chapter 4, an efficient palladium(II)-catalyzed chelation-controlled oxidative Heck type arylation reaction (Fujiwara-Moritani reaction) was performed between various arenes and trisubstituted olefins bearing a tether with a directing group (DG). Generally, intermolecular Heck type arylations did not work well for the synthesis of tetrasubstituted olefins due to the reluctance of trisubstituted olefins to participate in the carbopalladation process. However, an intermolecular arylation was successfully achieved by introducing a suitable directing group which could stabilize the palladium intermediate by chelation. In this way, we could synthesize various tetrasubstituted olefins in high yields. The Chapter 5 described the synthesis of poly-substituted tetrahydropyridines from Morita-Baylis-Hillman adducts modified with N-allylamino group via a radical cyclization process. An expedient method was developed for the synthesis of 1,4,5,6-tetrahydropyridines by radical cyclization protocol involving a consecutive 1,5-hydrogen transfer and a double bond isomerization process starting from the Morita-Baylis-Hillman adducts. The required starting materials were prepared from the Morita-Baylis-Hillman acetate of 2-bromobenzaldehyde as follows: (i) introduction of tosylamide moiety at the primary position and (ii) N-allylation with allyl bromide. The radical cyclization was carried out in benzene with n-Bu3SnH in the presence of AIBN. The protocol could be efficiently applied for the synthesis of antidepressant drug paroxetine and its analogs. In Chapter 6, a regioselective synthesis of poly-substituted thiophenes from Morita-Baylis-Hillman adducts is described. The reaction of Morita-Baylis-Hillman acetates and ethyl mercaptoacetate in the presence of DBU in DMF produced 2,3,4-trisubstituted tetrahydrothiophenes at room temperature as a diastereomeric mixture via the sequential SN2’ and Michael addition. Aromatization of tetrahydrothiophenes by DDQ oxidation produced 2,3,4-trisubstituted thiophenes in good yields. This is the first example on the synthesis of thiophene derivatives from the Morita-Baylis-Hillman adducts although there have been reported numerous syntheses of various cyclic and acyclic compounds by chemical transformation of the MBH adducts. In Chapter 7, an expedient synthesis of indolo[1,2-a]quinolines via Mn(OAc)3-mediated oxidative free radical cyclization and NaI/O2-assisted concomitant dealkoxycarbonylation/aerobic oxidation cascade is described. In a continuous study for the synthesis of indole-fused benzazepine derivatives (Chapter 2), the synthesis of indole-fused dihydroquinolines was examined. However, we have to use methyl 2-iodobenzoate as a starting material instead of the Morita-Baylis-Hillman adduct. An expedient synthetic procedure of indole[1,2-a]quinolines was developed using a sequential Cu-mediated N-arylation of indole, Mn(OAc)3-mediated oxidative free radical cyclization, and NaI/O2-assisted concomitant dealkoxycarbonylation/aerobic oxidation. The last step was replaced by a palladium-catalyzed decarboxylation/elimination protocol for the allyl ester derivatives. In Chapter 8, a direct one-pot introduction of 2-methylpyridines to Morita-Baylis-Hillman adducts via a base-mediated 3-aza-Cope rearrangement is described. An introduction of various nucleophilic components at the secondary position of the Morita-Baylis-Hillman adducts is very important in this chemistry. The introduction was carried out most frequently by the reaction of a nucleophile and a DABCO salt of the MBH adduct. Thus, the reaction failed when the nucleophile do not have an acidic proton. In these contexts, an introduction of 2-methylpyridine is almost impossible until now. Now we developed an easy route for the one-pot direct introduction of 2-methylpyridines at the secondary-position of a MBH adducts, that is the simple reaction of MBH bromide and 2-methylpyridine in acetonitrile in the presence of K2CO3. In the reaction, 2-methylpyridinium salt of MBH bromide was generated initially. A base treatment of the salt generated the corresponding N-allylenamine intermediate. The intermediate undergoes a facile 3-aza-Cope rearrangement to produce 2-pyridylmethyl-substituted MBH adduct, regioselectively. In the last Chapter 9, some additional results that are highly related to the above main contents of this thesis are described. Firstly, a Pd-catalyzed acetoxylation of uracil via an electrophilic palladation is described. Based on the biological importance of 5-substituted uracil derivatives, we examined some examples for the functionalization of a uracil ring. A palladium-catalyzed electrophilic acetoxylation at the electron-rich 5-position of uracil was carried out. The reaction proceeded via the electrophilic palladation of PdII at the 5-position of uracil ring, and regeneration of PdII was efficiently occurred with the aid of phenyliodonium diacetate (PIDA). As a second result, a palladium-catalyzed domino Heck-cyanation cascade to 3-cyanomethyl 2,3-dihydrobenzofuran and 2,3-dihydroindoles is described. An efficient synthesis of 3-methyl-3-cyanomethyl-2,3-dihydrobenzofuran and 2,3-dihydroindole derivatives was performed. The reaction was carried out from 2-bromophenyl methallyl ether or N-methallyl 2-bromoanilines via a domino Heck-cyanation process in moderate yields. As a cyanide source K4Fe(CN)6 was used. As a third extra result, a facile synthesis of cinnamamides from aromatic aldehydes and acetonitrile with Me3SiOK is described. We found that the use of potassium trimethylsilanolate is sufficient to deprotonate the weakly acidic acetonitrile and produced the Knovenagel condensation product between benzaldehyde and acetonitrile. In addition, a partial hydrolysis of cinnamonitrile to cinnamamide occurred simultaneously under the same conditions and eventually provided an easy route for the synthesis of various cinnamamide derivatives. As described in this thesis a variety of chemical transformations for the synthesis of poly-fused heterocyclic compounds, suitably modified MBH adducts, and various useful synthetic intermediates in organic synthesis have been investigated starting from the Morita-Baylis-Hillman adducts.

N2-Guanine DNA 손상이 인체 DNA polymerase ε catalytic subunit의 DNA 합성에 미치는 영향 연구

송인실 성균관대학교 일반대학원 2014 국내석사

진핵생물의 염색체 DNA 복제에 관여하는 replicative DNA polymerase(pol)에는 pol α, pol δ, pol ε이 속한다. 그 중 pol δ와 pol ε는 3’-5’ proofreading exonuclease 활성이 있기 때문에 DNA 복제 시 잘못 합성된 DNA 염기 서열을 교정하여 정확성을 훨씬 더 향상시킨다. 이중가닥으로 이루어진 DNA 중에서 pol δ는 lagging strand 합성에 주요 polymerase로 작용하며, pol ε는 leading strand 합성에 주요 polymerase로 작용한다. 그 중 본 연구에서는 인체 내 pol ε의 촉매 기능 탐색에 중점을 두고자 하였다. 인체 pol ε는 4개의 subunit, 즉 p261(POLE1), p59(POLE2), p17(POLE3), p12(POLE4)로 구성되어 있으며, 그 중에서도 POLE1이 촉매작용 역할을 한다고 알려져 있다. 인체 pol ε은 염색체 DNA 복제 시 leading strand 합성에 주로 관여하므로 정확하고 효율적인 복제가 이루어져야 한다. 그러나 내인적, 외인적으로 생성되는 많은 수의 DNA 손상물은 DNA 복제 시 DNA 삽입 오류 또는 정지를 유발하고 그로 인해 변이, 암 및 세포사 등을 초래하게 된다. 특히 인체 내 DNA guanine 염기의 N2 원소 위치에는 formaldehyde, acetaldehyde, styrene oxide, heterocyclic amines 또는 polycyclic aromatic hydrocarbons(e.g. benzo[a]pyrene)등의 발암원들에 의해 다양한 종류의 N2-G adduct들이 형성된다. 아직까지 우리 몸에서 흔히 생성되는 N2-G adduct들이 인체의 주요 replicative polymerase인 pol ε의 DNA 합성에 어떠한 영향을 미치는지에 대한 연구가 보고되어 있지 않다. 이에 착안하여 본 연구에서는 인체 pol ε catalytic subnuit의 N-말단 (1-1189 아미노산:pol ε1-1189) 부분을 이용하여 다양한 N2-G adduct들이 인체 pol ε의 DNA 합성시의 정확성 및 효율에 미치는 영향에 대하여 체계적으로 알아보고자 하였다. 이때 사용된 N2-G adduct들로는 크기가 작은 adduct인 N2-methyl(Me)G부터 N2-ethyl(Et)G, N2-isobutyl(Ib)G, N2-benzyl(Bz)G, N2-CH2-naphthyl(Naph)G에 이르기까지 점차 크기가 증가하는 N2-G adduct들을 사용하였다. 우선 exonuclease 활성이 제거된 pol ε1-1189을 E.coli에서 발현 후 정제하여 약 136 kDa에 달하는 단백을 분리하였으며, 이를 이용하여 primer extension, steady-state kinetics 등의 실험을 진행하였다. 먼저 full-length primer extension 실험 결과, 정상 G에 비해 N2-EtG 이상 크기의 손상에 대해서는 N2-adduct의 크기가 증가할수록 DNA 합성이 현저히 저해되고, 한 개의 염기만 삽입되며 합성이 정지됨을 정성적으로 확인할 수 있었다. 이를 정량적으로 알아보기 위하여 시행한 steady-state kinetics 분석 결과, 정확한 dNTP인 dCTP의 삽입 시의 kcat/Km(촉매 효율의 지표)값이 정상 G에 비해서 N2-MeG의 경우 120배, N2-EtG의 경우 5,000배, N2-IbG의 경우 약 10,000배 정도 adduct 크기에 비례하여 급격하게 감소함을 보였다. 이처럼 정확한 dCTP 삽입효율은 N2-G adduct 크기가 증가할수록 감소하는 반면, 잘못된 dATP의 오삽입비(misinsertion ratio)는 정상 G와 비교하여 N2-MeG의 경우 130배, N2-EtG의 경우 약 4,000배, N2-IbG의 경우는 7,400배까지 adduct 크기와 비례하여 증가하였으며, dTTP의 오삽입 역시 정상 G에 비해 N2-MeG는 8배, N2-EtG는 150배, N2-IbG는 5,000배의 증가를 보였다. 또한 정상 G 또는 N2-G adduct : C 염기쌍으로부터 다음 염기 extension시의 steady-state kinetics 분석을 통해서 kcat/Km 값을 조사한 결과, 정상 G에 비하여 N2-MeG는 90배, N2-EtG는 520배, N2-IbG는 5,500배 정도 감소하였다. 이상으로 본 연구에서는 N2-Guanine DNA adduct가 인체 pol ε의 손상물 맞은편 삽입반응과 후속적인 연장반응 단계에서 DNA 합성의 효율과 정확성을 adduct 크기에 의존적으로 저해함을 규명하였다. DNA polymerase(pol) α, pol δ and pol ε belong to replicative DNA polymerase that DNA synthesize chromosomal DNA in Eukaryote. Pol δ and pol ε have the very high fidelity in DNA replication, because they posess a 3’-5’ proofreading exonuclease activity. Pol δ is supposed to be a replicative polymerase mainly involved in lagging-strand DNA synthesis, while pol ε is supposed to be a replicative polymerase involved in leading-strand DNA synthesis. This study focuses on catalytic functions of human pol ε. Human pol ε consists of 4 subunits, p261(POLE1), p59(POLE2), p17(POLE3) and p12(POLE4). One of them, POLE1 has known to have the catalytic activity. During DNA replication, human pol ε participating in leading-strand DNA synthesis should replicate DNA very accurately and efficiently. However, a lot of DNA lesions generated from endogeneous, exogeneous sources cause replication error or blockage, which causes cancer, mutation and cell death. Especially guanine N2 atom in DNA is frequently modified by many carcinogens such as formaldehyde, acetaldehyde, styrene oxide, heterocyclic aromatic amines or polycyclic aromatic hydrocarbons(e.g. benzo[a]pyrene) to form various N2-G adducts. Effects of N2-G DNA adducts on DNA synthesis of human pol ε have not been studied yet. Therefore, in the present study the fidelity and catalytic efficiency for DNA synthesis against N2-G adducts were investigated using a recombinant human pol ε catalytic subunit(1-1189 amino acids) with DNA containing N2-G adducts from N2-methyl(Me)G, N2-ethyl(Et)G, N2-isobutyl(Ib)G, N2-benzyl(Bz)G, and N2-CH2-naphthyl(Naph)G. First, the recombinant exonuclease-free pol ε1-1189(136 kDa) were purified from induced Escherichia coli cells, and thereafter the primer extension and steady-state kinetics experiments were performed. Primer extension by pol ε were strongly blocked by N2-EtG and large adducts, which yielded only one base extended products. The steady-state kinetics parameter (kcat/Km) for dCTP incorporation by pol ε was reduced about 120-fold opposite N2-MeG, 5,000-fold opposite N2-EtG and 10,000-fold opposite N2-IbG, when compared to G, indicating that the catalytic efficiency of correct dCTP insertion proportionally decreases depending on the size of N2-G adducts. Misinsertion frequency for dATP increases about 130-fold opposite N2-MeG, 4,000-fold opposite N2-EtG, and 7,400-fold opposite N2-IbG, when compared to G. Misinsertion frequency for dTTP increased 8-fold opposite N2-MeG, 150-fold opposite N2-EtG and 5,000-fold opposite N2-IbG, when compared to G. Furthermore, steady-state kinetic parameters(kcat/Km) for next base extension from G (or N2-G adducts): C pair by pol ε were also 90-fold following N2-MeG :C, 520-fold N2-EtG:C, 5,500-fold following N2-IbG:C, when compared to G:C pair. This results suggest that N2-guanine DNA adducts interfere with the function of human pol ε for DNA synthesis at the insertion step opposite the lesion and the next extension step, in terms of fidelity and catalytic efficiency, in the adduct-bulk dependent manner.