, 람 세이 기념 트러스트 (연구 활동 JAB에 2,009에서 2,011 사이), 임페리얼 칼리지 런던 대학, 재정 지원을 위해 우리는 기꺼이 EPSRC (EP/J001538/1 JAB에 경력 가속 친목을) 인정합니다. 관대 한 지원과 조언을 교수 앨런 암스트롱을 부탁드립니다.

Diiodomethyllithium와 Iodoaziridines 1. 준비 <ol><li> 불꽃은 아르곤 분위기 하에서 실온으로 냉각되도록 한 후, 아르곤 기류하에 교반기 바 및 셉텀이 장착을 함유 한 100 ㎖ 둥근 바닥 플라스크를 건조. 주 : 유리는 오븐에서 밤새 (125 ° C)에서 건조와 유사한 방식으로 실내 온도로 냉각도 적합합니다. </li><li> 플라스크에, 또는 마이크로 통해 5.7 ㎖의 무수 THF를 추가하고 2.7 ㎖의 무수 잇 2 O 주사기를 통해 갓 증류 헥사 메틸 디 실라 잔 (1.50 밀리몰, 315 μL). </li><li> 플라스크 잘 빠져들 수 있도록 적절한 크기 듀어에서 드라이 아이스 / 아세톤 조에서 -78 ° C의 생성 된 용액을 냉각 및 교반 하였다. 빛에 반응 용기의 노출을 최소화하기 위해, 알루미늄 호일로 듀어 커버. </li><li> -78 ℃의 용액에 2 ~ 3 분에 걸쳐 주사기를 통해 적가 N BuLi를 (1.50 밀리몰, 0.60 ㎖, 헥산 2.5 M)혼합물 LiHMDS의 0.17 M 용액을 형성하기 위해, 또 30 분 동안 -78 ℃에서 교반하는 것을 허용한다. 주의 : N BuLi를 솔루션은 가연성 피부에 부식성 및 자연 발화성이다. 주사기 초과 시약 따라 침묵해야한다. </li><li> 30 분 후, 또는 마이크로를 통해 디 요오도 (1.70 밀리몰, 135 μL) 다음에 주사기를 통해 불꽃 - 건조 된 둥근 바닥 플라스크에 10 ㎖, 1 ㎖의 무수 THF를 추가하고 잘 혼합되어 있는지 확인합니다. </li><li> -78 ℃에서 리튬 헥사 메틸 디 실라 잔의 용액을 2 분에 걸쳐 디 요오 용액 적하 추가 -78 ° C에서 20 분 동안이 솔루션을 남겨주세요 </li><li> 또 다른 라운드 불꽃 건조 10 ㎖ 바닥 플라스크에 [(E)-4-methylphenylmethylidene] -4 - 메틸 벤젠 (137 ㎎, 0.50 mmol) 및 2.0 ㎖의 무수 THF에 용해 -이 시간 동안, N을 단다. </li><li> 20 분 탈 양성자 시간 후, -78 및 #에서 5 분에 걸쳐 diiodomethyllithium 용액에 이민 용액을 적가176, C. </li><li> 적가가 완료된 직후, 드라이 아이스 목욕에서 반응 용기를 들어, 0 ℃의 얼음 / 물을 욕조로 전송 다시 커버를 알루미늄 호일로 0 ℃에서 15 분 방치 참고 :이 솔루션은 컬러에 오렌지 있어야한다. </li><li> 0 ° C에서 15 분 후, 30 ㎖의 포화 중탄산 나트륨 수용액을 첨가하여 반응을 급냉. 분액 깔때기에 혼합물을 전송하고 30 ㎖의 CH 2를 망할 CIA 2를 추가합니다. 혼합물을 흔들어 하단을 CH2Cl2 층을 제거. 이 추출 과정을 두 번 더 반복하고, CH 2 망할 CIA 2 층을 결합한다. </li><li> 용액에 존재하는 모든 물을 제거하는 유기층에 황산나트륨을 추가 한 후, 황산나트륨을 필터링하고 둥근 바닥 플라스크에 250 ml의 여액을 모은다. </li><li> 원하는 iodoaziridine 상품의 불순한 샘플을 수득하는 회전 증발기상에서 감압하에 용매를 제거. </li></ol> 크로마토 그래피에 대한 고정 단계로 제품의 안정성 2. 평가 <ol><li> 을 CH2Cl2 (16 ㎖)의 원유 아지리 샘플을 녹이고이 완전히 용해 보장, 내부 표준 (28.0 ㎎, 0.167 mmol)을 1,3,5 - 트리 메 톡시 벤젠을 추가합니다. 이 혼합물로부터 분취 량 (2 ml)에 걸리는, 감압하에 용매를 제거하고 하나의 H NMR 스펙트럼에 의해이 샘플을 분석한다. </li><li> 표준 NMR 처리 소프트웨어를 사용하여 기록 된 하나의 H NMR 스펙트럼을 연다. Mestrenova에서 마우스 오른쪽 통합 도구를 제공하기 위해, 다음 &quot;수동&quot;스펙트럼을 클릭하고 &quot;통합&quot;을 선택했다. 각각 내부 표준 및 아지리 딘 CHI 신호의 신호를 통합하는 6.08 ppm 이하 및 4.87 PPM에서 피크의 폭을 충당하기 위해 클릭하고 드래그. 6.08 PPM에서의 피크의 통합을 마우스 오른쪽 버튼으로 클릭, &quot;통합 편집&quot;을 선택하고 3.0 &quot;정규화&quot;값을 변경합니다. NOTE : 비슷한 단계는 다른 소프트웨어 패키지에 적용 할 수 있습니다. </li><li> (3분의 100) × 59 %의 계산 된 수익률을 준다 (CHI 신호의 정수)를 사용하여 여기에, iodoaziridine의 수율을 결정하는 아지리 딘 CHI 신호 (4.87 ppm)를위한 통합의 업데이트 된 값을 사용합니다. NOTE : 내부 표준 (0.167 밀리몰), 및 하나의 양성자에 해당 상품 피크의 알려진 양이 주어 iodoaziridine의 수율은 다음의 수학 식에 의해 계산된다 : 100 × (제품 피크의 적분) × (내부 표준 물질의 몰수) / 몰 출발 물질. </li><li> 각 5 %의 EtOAc / 헥산에게 실리카, 실리카 +1 % 순 3 (트리 에틸 아민), 중성 알루미나, 기본 알루미나 (활동 I), 기본 알루미나 (활동 IV)와 플로리, 다음 고정상 (25g)의 슬러리를 준비 교반 막대를 함유하는 여섯 별도의 250 ㎖의 삼각 플라스크 (50 ㎖). 다른 원추형 플라스크에 대조 실험으로 사용될, 5 %의 EtOAc / 헥산 용액 (50 ㎖)을 제조. 주의 : silic고용 겔, 알루미나 및 기타 고정 단계 흡입하면, 따라서 항상 효과적인 흄 후드에서 처리해야한다 위험합니다. </li><li> RT의 원추형 플라스크 각각에 iodoaziridine / 내부 표준 용액 2 ㎖를 분취를 추가합니다. 30 분 동안 슬러리 혼합물을 교반 하였다. 주 :이 화합물은 통상 컬럼 크로마토 그래피 절차 중에 정지상에 노출 될 수있는 기간을 나타낸다. </li><li> 소결 깔때기를 사용하여 슬러리 혼합물을 필터링하고 둥근 바닥 플라스크 250 ㎖에 여과 액을 수집합니다. CH 2 망할 CIA 2 (2 × 30 ㎖)으로 소결 깔때기에 잔류 물을 씻는다. 나머지 슬러리이 필터링 과정을 반복합니다. 주 : 여과 시간을 허용하고 있으므로 정지상 물질 각각에 대해 동일한 시간을 유지하기 위해 각 정지상의 개시를 상쇄하는 것이 적절하다. </li><li> 감압하에 생성 된 샘플로부터 용매를 제거하고, 1 H NMR 분광 분석에 의해2.2 절에 설명 된대로 iodoaziridine은, 각 경우에서 회수의 양을 계산하는 SCOPY. </li><li> 2.1 절에서 얻은 그와 함께 각 시험 정지상에서 얻은 iodoaziridine의 수율을 비교. 참고 : 가장 높은 수익률 2.1 이상적으로 동일한주는 샘플은 크로마토 그래피에 대한 최적의 고정상을 나타냅니다. 이 예에서는 기본 알루미나 (활동 IV)는 정제를 위해 최선의 고정상을 간주되었다. </li></ol> Iodoaziridine의 기본 알루미나와 정제 3. 비활성화 <ol><li> 조 iodoaziridine 혼합물을 생성하는 제 1 절을 반복합니다. </li><li> 기본 알루미나 (활동 IV)를 생성하기 위해, 500 ㎖의 둥근 바닥 플라스크에 기본 알루미나 (활동 I)의 100g을 추가 한 다음 플라스크에 물 10 ㎖를 추가하고 유리 마개에 맞게. </li><li> 아무 덩어리도 알루미나를 통해 물 확산을 나타내는 볼 수 없을 때까지 적극적으로 플라스크를 흔들어. 알루미나는 실온으로 냉각 할 수 있습니다. 주의 : adsorp물 기는 발열, 그래서 플라스크 뜨거운받을 수 있습니다 및 압력의 축적이 발생할 수 있습니다. 모든 압력이 자주 릴리스 빌드. </li><li> 5 %의 EtOAc / 헥산으로 그레이딩, 헥산으로 용리, 정지상으로서 염기성 알루미나 (활성 IV)를 사용하여 컬럼 크로마토 그래피에 의해 조질 iodoaziridine 정화. 참고 : EtOAc로 높은 농도의 기본 알루미나와 함께 사용할 수 없습니다. 이러한 경우, 디 에틸 에테르 대신에 사용될 수있다. </li><li> 생성물 함유 분획을 합하여 순수한 iodoaziridine을 감압 하에서 용매를 제거한다. </li></ol>

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+cis-selective+synthesis+of+iodo-aziridines+using+diiodomethyllithium+and+in+situ+generated+N-Boc-imines.">Highly cis-selective synthesis of iodo-aziridines using diiodomethyllithium and in situ generated N-Boc-imines.</a> Chem. Commun. 48 (100), 12246-12248 (2012).</li><li>Boultwood, T., Affron, D. P., Trowbridge, A. D., Bull, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+cis-C-iodo-N-tosyl-aziridines+using+diiodomethyllithium:+reaction+optimization,+product+scope+and+stability,+and+a+protocol+for+selection+of+stationary+phase+for+chromatography.">Synthesis of cis-C-iodo-N-tosyl-aziridines using diiodomethyllithium: reaction optimization, product scope and stability, and a protocol for selection of stationary phase for chromatography.</a> J. Org. Chem. 78 (13), 6632-6647 (2013).</li><li>Bull, J. A., Charette, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+procedure+for+the+synthesis+of+gem-diiodoalkanes+by+the+alkylation+of+diiodomethane.+scope+and+limitations.">Improved procedure for the synthesis of gem-diiodoalkanes by the alkylation of diiodomethane. scope and limitations.</a> J. Org. Chem. 73 (20), 8097-8100 (2008).</li><li>Bull, J. A., Charette, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intramolecular+Simmons-Smith+cyclopropanation.+Studies+into+the+reactivity+of+alkyl-substituted+zinc+carbenoids,+effect+of+directing+groups+and+synthesis+of+bicyclo[n.1.0]alkanes.">Intramolecular Simmons-Smith cyclopropanation. Studies into the reactivity of alkyl-substituted zinc carbenoids, effect of directing groups and synthesis of bicyclo[n.1.0]alkanes.</a> J. Am. Chem. Soc. 132 (6), 1895-1902 (2010).</li><li>Lim, D. S. W., Anderson, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One-step+preparation+of+functionalized+(E)-vinylsilanes+from+aldehydes.">One-step preparation of functionalized (E)-vinylsilanes from aldehydes.</a> Org. Lett. 13 (18), 4806-4809 (2011).</li><li>Bull, J. A., Mousseau, J. J., Charette, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Convenient+one-pot+synthesis+of+(E)-&#946;-aryl+vinyl+halides+from+benzyl+bromides+and+dihalomethanes.">Convenient one-pot synthesis of (E)-&#946;-aryl vinyl halides from benzyl bromides and dihalomethanes.</a> Org. Lett. 10 (23), 5485-5488 (2008).</li><li>Bull, J. A., Mousseau, J. J., Charette, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+(E)-(2-iodovinyl)benzene+from+benzyl+bromide+and+diiodomethane.">Preparation of (E)-(2-iodovinyl)benzene from benzyl bromide and diiodomethane.</a> Org. Synth. 87, 170-177 (2010).</li><li>Boxer, M. 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The authors declare that they have no competing financial interests.

CIS의 입체 이성질체 제조 과정 - N-TS-iodoaziridines 정량적 플래시 칼럼 크로마토 그래피에 의해 잠재적으로 불안정한 화합물의 정제를위한 최선의 정지상을 나타내는 안정성 시험 프로토콜과 함께 설명한다. 그것은 이러한 접근법을 통해 iodoaziridines 액세스가 그대로 링의 유도체 화에 의해 개발 될 아지리 딘의 다양한 액세스 방법을 가능하게 할 것이라고 예상된다. <p class="jove_conten...

이 방법의 목적은 아지리 딘 유도체에 더 작용에 대한 가능성을 제공 iodoaziridines을 준비하는 것입니다. 방법은 크로마토 그래피를위한 정지상 최적의 정량적 선택을위한 프로토콜을 포함한다. 아지리 딘, 유기 화학 1에서 그 중요한 빌딩 블록을 만드는 세 가지 원환, 민병대 고유 링 변형 등. 그들은 특히 작용 아민 4,5의 합성 중간체 자주 아지리 링 오프닝 2,3 관련된 반응의 광대 한 배열, 또는 다른 질소 함유 헤테로 고리 6,7의 형성을 표시합니다. 그대로 아지리 딘 고리를 포함하는 전구체의 작용에 의해 아지리 딘 유도체의 범위의 합성은 실행 가능한 전략 8로 떠오르고있다. 기능 그룹 금속 교환, 아지 리디 음이온을 생성하고, 친전 자체와 반응이 효과적인 것으로 나타났다 <suP&gt; 9,10,11 및 N-보호 아지리 딘의 최근 레기 및 입체 탈 양성자는 12-15을 달성하고있다. 아주 최근에, 팔라듐 Vedejs (16, 17)에 의해 개발 된 기능화 아지리 전구체에서 아릴 아지리 딘을 형성하는 크로스 커플 링 방법을 촉매하고, 자신 18. 헤테로 원자 치환 된 아지리 딘의 화학 반응성 및 안정성 (19)의 매혹적인 질문을 엽니 다. 우리는 기존 아지리 딘 작용 화 반응에 상보 반응성 유도체의 광범위한 전구체를 제공하는 잠재력을 제공 신규 관능기로서 iodoaziridines의 제조에서 관심이되고있다. 2012 년 우리는 아릴 N-BOC-iodoaziridines (20)의 첫 번째 준비를보고, 아주 최근에 N-TS-iodoaziridines 21 치환 된 아릴 및 알킬의 준비를보고했다. ACCE에 대한 방법SS 정보가 iodoaziridines diiodomethyllithium를 사용하여, 또한 최근 diiodoalkanes 22, 23의 제조에 사용 된 시약, 22, 24을 diiodomethylsilanes 및 비닐 요오드 25-27. 이 시약의 carbenoid 같은 특성은 낮은 온도 (22,28)의 준비와 사용이 필요합니다. iodoaziridines의 준비 diiodomethyllithium의 생성에 사용되는 기술과 조건은 다음과 같다. 실리카 크로마토 그래피 (29)에 대한 선택의 재료로 등장했다 동안, N-TS-iodoaziridines의 정화에 적합하지 않은 것으로 판명. 실리카 겔은 일반적으로 인해 가능하고 효과적인 분리에 유기 화학에서 플래쉬 크로마토 그래피에 사용되는 첫번째 및 고상 물질이다. 그러나, 실리카 겔, 산성 성질은 원하는 물질의 분리를 방지하는 정제 중에 민감한 기질의 분해를 일으킬 수있다. 동안 다른 성ationary 단계 또는 수정 실리카 젤은 크로마토 그래피 (30)에 사용할 수있는, 서로 다른 재료로 표적 분자의 호환성을 평가하는 방법이 없었습니다. 때문에 iodoaziridines의 민감한, 우리는 여기에서 입증되고 고정상 21 일의 배열 화합물의 안정성을 평가하기 위해 프로토콜을 설립했다. 이것은 민감한 작용기를 가진 화합물의 폭 넓은 범위의 합성에 응용 프로그램에 대한 잠재력을 가지고 있습니다. 다음 프로토콜은 높은 수율 알킬 및 방향족 시스 iodoaziridines 모두의 입체 이성질체 합성을 허용, N-TS의 iodoaziridines에 대한 효율적인 액세스를 제공합니다.

<table border="0" cellpadding="0" cellspacing="0" width="871">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:209px;">Hexamethyldisilazane</td>
			<td style="width:273px;">999-97-3</td>
			<td style="width:197px;">Alfa Aesar</td>
			<td style="width:192px;">Distill from KOH under argon prior to use.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:209px;">n-Butyllithium</td>
			<td style="width:273px;">109-72-8</td>
			<td style="width:197px;">Sigma Aldrich</td>
			<td style="width:192px;">2.5 M in hexanes, titrate prior to use.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:209px;">Diiodomethane</td>
			<td style="width:273px;">75-11-6</td>
			<td style="width:197px;">Alfa Aesar</td>
			<td style="width:192px;">Contains copper as a stabilizer.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">1,3,5-Trimethoxybenzene</td>
			<td style="width:273px;">621-23-8</td>
			<td style="width:197px;">Sigma Aldrich</td>
			<td style="width:192px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">Silica</td>
			<td style="width:273px;">112945-52-5</td>
			<td style="width:197px;">Merck</td>
			<td style="width:192px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">Basic alumina</td>
			<td style="width:273px;">1344-28-1</td>
			<td style="width:197px;">Sigma Aldrich</td>
			<td style="width:192px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">Neutral alumina</td>
			<td style="width:273px;">1344-28-1</td>
			<td style="width:197px;">Merck</td>
			<td style="width:192px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">Florisil</td>
			<td style="width:273px;">1343-88-0</td>
			<td style="width:197px;">Sigma Aldrich</td>
			<td style="width:192px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:209px;">THF</td>
			<td colspan="3" rowspan="3" style="width:663px;">All anhydrous solvents were dried through activated alumina purification columns.&#160;</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:209px;">Et2O</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:209px;">CH2Cl2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NMR spectrometer</td>
			<td>Bruker AV 400&#160;</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:209px;">NMR processing software</td>
			<td>MestReNova&#160;</td>
			<td>7.0.2-8636</td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis and purification of iodoaziridines involving quantitative selection of the optimal stationary phase for chromatography

시스의 높은 부분 입체 준비 - N-TS aldimines와 diiodomethyllithium의 반응을 통해 N-TS-iodoaziridines 설명한다. Diiodomethyllithium는 THF / 디 에틸 에테르의 혼합물에, -78 ° C에서, LiHMDS와 디 요오의 탈 양성자에 의해 준비가되어 있습니다 어둠 속에서. 이러한 조건은 생성 LiCHI이 시약의 안정성을 위해 필수적이다. 예비 성형 diiodomethyllithium 용액에 N-TS aldimines의 후속 적가 절연되어 있지 않은, 아미노 - 디요오다 이드 중간을 제공한다. 0 ° C에 반응 혼합물의 급속한 온난화 전용 시스 - 부분 입체 선택성으로 iodoaziridines을 감당할 고리 화 반응을 촉진합니다. 반응물의 첨가 및 고리 화 반응 단계는 세세한 온도 제어로 한 반응 플라스크에 중재된다. 때문에 정화, PU의 적절한 방법의 평가에 iodoaziridines의 감도rification가 필요합니다. 컬럼 크로마토 그래피를위한 정지상에 민감한 화합물의 안정성을 평가하기위한 프로토콜을 설명한다. 이 방법은 새로운 iodoaziridines, 또는 기타 잠재적으로 중요한 신규 화합물에 적용하기에 적합하다. 따라서이 방법은 합성 프로젝트의 범위에서 응용 프로그램을 찾을 수 있습니다. 절차는 먼저 내부 표준에 비해 1 H의 NMR 분광법의 사전 정제에 대한 반응 수율의 평가를 포함한다. 불순한 생성물 혼합물의 일부는 다음 플래시 크로마토 그래피 용 리제로서 적합한 용매 시스템에서, 크로마토 그래피에 적합한 다양한 정지상의 슬러리에 노출된다. 다음에 여과 크로마토 모방 30 분간, 교반 한 후, 샘플을 1 H NMR 분광기에 의해 분석된다. 각 정지상 계산 수율이어서 초기 조질 반응 혼합물로부터 얻어지는 것과 비교된다. 얻어진 결과는 t의 양적 평가를 제공다른 고정 단계에 화합물 그는 안정성; 따라서 최적의 선택할 수 있습니다. 적합한 고정상 우수한 수율과 순도의 특정 iodoaziridines의 분리 허용되면서, 활동 IV에 수정 된 기본 알루미나의 선택.

(도 1) 단일 부분 입체 이성질체로서 및 우수한 순도 (4 - 톨 일설 포닐) 아지리 딘 - (±) -2 - 요오도 -3 - (4 - 톨릴) -1 - 프로 가르치시 시스를 설명했다. 종래 정제, iodoaziridine 상품의 59 %의 수율은 하나의 H NMR 스펙트럼에 의해 계산 하였다. 그러나,이 iodoaziridine은 정화하는 것이 특히 어려웠다 실리카에 상당한 분해를 시행 하였다. 정지상 화면에 의해 결정된 바와 같이 염?...

Watch this Scientific Journal Video about Synthesis and Purification of Iodoaziridines Involving Quantitative Selection of the Optimal Stationary Phase for Chromatography at JoVE.com

Synthesis and Purification of Iodoaziridines Involving Quantitative Selection of the Optimal Stationary Phase for Chromatography

시스의 입체 이성질체 한 냄비 준비를위한 프로토콜 - N-TS-iodoaziridines 설명한다. N-TS aldimines과 보석 - 디요오다 이드 iodoaziridines에 중간 아미노산이 입증되고의 고리 화 반응에 diiodomethyllithium, 추가의 생성. 또한 신속하고 정량적 크로마토 그래피에 가장 적합한 고정상을 평가하는 프로토콜이 포함된다.

합성 및 크로마토 그래피에 대한 최적의 고정상의 양이 선택의 참여 Iodoaziridines의 정화

The highly diastereoselective preparation of cis-N-Ts-iodoaziridines through reaction of diiodomethyllithium with N-Ts aldimines is described. ...

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JoVE Journal

Chemistry

12.4K 조회수.  Imperial College London.  시스의 입체 이성질체 한 냄비 준비를위한 프로토콜 - N-TS-iodoaziridines 설명한다. N-TS aldimines과 보석 - 디요오다 이드 iodoaziridines에 중간 아미노산이 입증되고의 고리 화 반응에 diiodomethyllithium, 추가의 생성. 또한 신속하고 정량적 크로마토 그래피에 가장 적합한 고정상을 평가하는 프로토콜이 포함된다. 

비디오: Synthesis and Purification of Iodoaziridines Involving Quantitative Selection of the Optimal Stationary Phase for Chromatography

Determination of the Gas-phase Acidities of Oligopeptides

Amino acid residues located at different positions in folded proteins often exhibit different degrees of acidities. For example, a cysteine residue located at or near the N-terminus of a helix is often more acidic than that at or near the C-terminus 1-6. Although extensive experimental studies on the acid-base properties of peptides have been carried out in the condensed phase, in particular in aqueous solutions 6-8, the results are often complicated by solvent effects 7. In fact, most of the active sites in proteins are located near the interior region where solvent effects have been minimized 9,10. In order to understand intrinsic acid-base properties of peptides and proteins, it is important to perform the studies in a solvent-free environment.
We present a method to measure the acidities of oligopeptides in the gas-phase. We use a cysteine-containing oligopeptide, Ala3CysNH2 (A3CH), as the model compound. The measurements are based on the well-established extended Cooks kinetic method (Figure 1) 11-16. The experiments are carried out using a triple-quadrupole mass spectrometer interfaced with an electrospray ionization (ESI) ion source (Figure 2). For each peptide sample, several reference acids are selected. The reference acids are structurally similar organic compounds with known gas-phase acidities. A solution of the mixture of the peptide and a reference acid is introduced into the mass spectrometer, and a gas-phase proton-bound anionic cluster of peptide-reference acid is formed. The proton-bound cluster is mass isolated and subsequently fragmented via collision-induced dissociation (CID) experiments. The resulting fragment ion abundances are analyzed using a relationship between the acidities and the cluster ion dissociation kinetics. The gas-phase acidity of the peptide is then obtained by linear regression of the thermo-kinetic plots 17,18.
The method can be applied to a variety of molecular systems, including organic compounds, amino acids and their derivatives, oligonucleotides, and oligopeptides. By comparing the gas-phase acidities measured experimentally with those values calculated for different conformers, conformational effects on the acidities can be evaluated.

The determination of the gas-phase acidities of cysteine-containing oligopeptides is described. The experiments are performed using a triple quadrupole mass spectrometer. The relative acidities of the peptides are measured using collision-induced dissociation experiments, and the quantitative acidities are determined using the extended Cooks kinetic method.

올리고 펩타이드의 가스 단계 산성도의 결정

Amino acid residues located at different positions in folded proteins often exhibit different degrees of acidities. For example, a cysteine residue ...

연구

화학

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid phase by using combinatorial chemistry. One of the most well studied classes of peptidomimetics is peptoids. Peptoids are easy to synthesize and have been shown to be proteolysis-resistant and cell-permeable. Over the past decade, many useful protein ligands have been identified through screening of peptoid libraries. However, most of the ligands identified from peptoid libraries do not display high affinity, with rare exceptions. This may be due, in part, to the lack of chiral centers and conformational constraints in peptoid molecules. Recently, we described a new synthetic route to access peptide tertiary amides (PTAs). PTAs are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. With side chains on both &#945;-carbon and main chain nitrogen atoms, the conformation of these molecules are greatly constrained by sterical hindrance and allylic 1,3 strain. (Figure 1) Our study suggests that these PTA molecules are highly structured in solution and can be used to identify protein ligands. We believe that these molecules can be a future source of high-affinity protein ligands. Here we describe the synthetic method combining the power of both split-and-pool and sub-monomer strategies to synthesize a sample one-bead one-compound (OBOC) library of PTAs.

Peptide tertiary amides (PTAs) are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. Here we describe a synthetic method which combines&#160;both split-and-pool and sub-monomer strategies to synthesize a one-bead one-compound library of PTAs.

분할 및 풀 합성 펩타이드 급 아미드 도서관의 특성

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

Quantitative Detection of Trace Explosive Vapors by Programmed Temperature Desorption Gas Chromatography-Electron Capture Detector

The direct liquid deposition of solution standards onto sorbent-filled thermal desorption tubes is used for the quantitative analysis of trace explosive vapor samples. The direct liquid deposition method yields a higher fidelity between the analysis of vapor samples and the analysis of solution standards than using separate injection methods for vapors and solutions, i.e., samples collected on vapor collection tubes and standards prepared in solution vials. Additionally, the method can account for instrumentation losses, which makes it ideal for minimizing variability and quantitative trace chemical detection. Gas chromatography with an electron capture detector is an instrumentation configuration sensitive to nitro-energetics, such as TNT and RDX, due to their relatively high electron affinity. However, vapor quantitation of these compounds is difficult without viable vapor standards. Thus, we eliminate the requirement for vapor standards by combining the sensitivity of the instrumentation with a direct liquid deposition protocol to analyze trace explosive vapor samples.

Trace explosive vapors of TNT and RDX collected on sorbent-filled thermal desorption tubes were analyzed using a programmed temperature desorption system coupled to GC with an electron capture detector. The instrumental analysis is combined with direct liquid deposition method to reduce sample variability and account for instrumentation drift and losses.

프로그램 된 온도 탈착 가스 크로마토 그래피 - 전자 캡처 검출기에 의해 추적 폭발성 증기의 양이 감지

The direct liquid deposition of solution standards onto sorbent-filled thermal desorption tubes is used for the quantitative analysis of trace explosive ...

Supercritical Nitrogen Processing for the Purification of Reactive Porous Materials

Supercritical fluid extraction and drying methods are well established in numerous applications for the synthesis and processing of porous materials. Herein, nitrogen is presented as a novel supercritical drying fluid for specialized applications such as in the processing of reactive porous materials, where carbon dioxide and other fluids are not appropriate due to their higher chemical reactivity. Nitrogen exhibits similar physical properties in the near-critical region of its phase diagram as compared to carbon dioxide: a widely tunable density up to ~1 g ml-1, modest critical pressure (3.4 MPa), and small molecular diameter of ~3.6 &#197;. The key to achieving a high solvation power of nitrogen is to apply a processing temperature in the range of 80-150 K, where the density of nitrogen is an order of magnitude higher than at similar pressures near ambient temperature. The detailed solvation properties of nitrogen, and especially its selectivity, across a wide range of common target species of extraction still require further investigation. Herein we describe a protocol for the supercritical nitrogen processing of porous magnesium borohydride.

Nitrogen is an effective supercritical fluid for extraction or drying processes due to its small molecular size, high density in the near-liquid supercritical regime, and chemical inertness. We present a supercritical nitrogen drying protocol for the purification treatment of reactive, porous materials.

반응성이 있음 다공성 물질의 정제를위한 초 임계 질소 처리

Supercritical fluid extraction and drying methods are well established in numerous applications for the synthesis and processing of porous materials. ...

A Quantitative Glycomics and Proteomics Combined Purification Strategy

There is a growing desire in the biological and clinical sciences to integrate and correlate multiple classes of biomolecules to unravel biology, define pathways, improve treatment, understand disease, and aid biomarker discovery. N-linked glycosylation is one of the most important and robust post-translational modifications on proteins and regulates critical cell functions such as signaling, adhesion, and enzymatic function. Analytical techniques to purify and analyze N-glycans have remained relatively static over the last decade. While accurate and effective, they commonly require significant expertise and resources. Though some high-throughput purification schemes have been developed, they have yet to find widespread adoption and often rely on the enrichment of glycopeptides. One promising method, developed by Thomas-Oates et al., filter aided N-glycan separation (FANGS), was qualitatively demonstrated on tissues. Herein, we adapted FANGS to plasma and coupled it to the individuality normalization when labeling with glycan hydrazide tags strategy in order to achieve accurate relative quantification by liquid chromatography mass spectrometry and enhanced electrospray ionization. Furthermore, we designed new functionality to the protocol by achieving tandem, shotgun proteomics and glycosylation site analysis on hen plasma. We showed that N-glycans purified on filter and derivatized by hydrophobic hydrazide tags were comparable in terms of abundance and class to those by solid phase extraction (SPE); the latter is considered a gold standard in the field. Importantly, the variability in the two protocols was not statistically different. Proteomic data that was collected in-line with glycomic data had the same depth compared to a standard trypsin digest. Peptide deamidation is minimized in the protocol, limiting non-specific deamidation detected at glycosylation motifs. This allowed for direct glycosylation site analysis, though the protocol can accommodate 18O site labeling as well. Overall, we demonstrated a new in-line high-throughput, unbiased, filter based protocol for quantitative glycomics and proteomics analysis.

A high-throughput protocol was developed for combined proteomics and glycomics purification and LC-MS/MS quantification in plasma. Deamidation analysis of N-linked glycosylation motifs was specific to deglycosylated sites. Accurate quantitation of N-glycans was achieved by coupling filter aided N-glycan separation to the individuality normalization when labeling with glycan hydrazide tags strategy.

양적 글리코 믹스와 단백질 체학 복합 정제 전략

There is a growing desire in the biological and clinical sciences to integrate and correlate multiple classes of biomolecules to unravel biology, define ...

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of trifluoromethoxylated aromatic compounds remains a formidable challenge in organic synthesis. Conventional approaches often suffer from poor substrate scope, or require use of highly toxic, difficult-to-handle, and/or thermally labile reagents. Herein, we report a user-friendly protocol for the synthesis of methyl 4-acetamido-3-(trifluoromethoxy)benzoate using 1-trifluoromethyl-1,2-benziodoxol-3(1H)-one (Togni reagent II). Treating methyl 4-(N-hydroxyacetamido)benzoate (1a) with Togni reagent II in the presence of a catalytic amount of cesium carbonate (Cs2CO3) in chloroform at RT afforded methyl 4-(N-(trifluoromethoxy)acetamido)benzoate (2a). This intermediate was then converted to the final product methyl 4-acetamido-3-(trifluoromethoxy)benzoate (3a) in nitromethane at 120 &#176;C. This procedure is general and can be applied to the synthesis of a broad spectrum of ortho-trifluoromethoxylated aniline derivatives, which could serve as useful synthetic building blocks for the discovery and development of new pharmaceuticals, agrochemicals, and functional materials.

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

An operationally simple procedure for the synthesis of ortho-trifluoromethoxylated aniline derivatives via a two-step sequence of O-trifluoromethylation of N-aryl-N-hydroxyacetamide followed by thermally induced intramolecular OCF3-migration is reported.

의 합성을위한 프로토콜<em&gt; 직교</em&gt; -trifluoromethoxylated 아닐린 파생 상품

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of ...

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the development of antibody drug conjugates (ADCs) and nanomedicine, fluorescent microscopy and systems chemistry. For many of these applications, specificity of the bioconjugation method used is of prime concern. The Michael addition of maleimides with cysteine(s) on the target proteins is highly selective and proceeds rapidly under mild conditions, making it one of the most popular methods for protein bioconjugation.
We demonstrate here the modification of the only surface-accessible cysteine residue on yeast cytochrome c with a ruthenium(II) bisterpyridine maleimide. The protein bioconjugation is verified by gel electrophoresis and purified by aqueous-based fast protein liquid chromatography in 27% yield of isolated protein material. Structural characterization with MALDI-TOF MS and UV-Vis is then used to verify that the bioconjugation is successful. The protocol shown here is easily applicable to other cysteine - maleimide coupling of proteins to other proteins, dyes, drugs or polymers.

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

This protocol details the important steps required for the bioconjugation of a cysteine containing protein to a maleimide, including reagent purification, reaction conditions, bioconjugate purification and bioconjugate characterization.

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the ...

Facile Synthesis of Worm-like Micelles by Visible Light Mediated Dispersion Polymerization Using Photoredox Catalyst

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins with the synthesis of a hydrophilic poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA) homopolymer using reversible addition-fragmentation chain-transfer (RAFT) polymerization. Under mild visible light irradiation (&#955; = 460 nm, 0.7 mW/cm2), this macro-chain transfer agent (macro-CTA) in the presence of a ruthenium based photoredox catalyst, Ru(bpy)3Cl2 can be chain extended with a second monomer to form a well-defined block copolymer in a process known as Photoinduced Electron Transfer RAFT (PET-RAFT). When PET-RAFT is used to chain extend POEGMA with benzyl methacrylate (BzMA) in ethanol (EtOH), polymeric nanoparticles with different morphologies are formed in situ according to a polymerization-induced self-assembly (PISA) mechanism. Self-assembly into nanoparticles presenting POEGMA chains at the corona and poly(benzyl methacrylate) (PBzMA) chains in the core occurs in situ due to the growing insolubility of the PBzMA block in ethanol. Interestingly, the formation of highly pure worm-like micelles can be readily monitored by observing the onset of a highly viscous gel in situ due to nanoparticle entanglements occurring during the polymerization. This process thereby allows for a more reproducible synthesis of worm-like micelles simply by monitoring the solution viscosity during the course of the polymerization. In addition, the light stimulus can be intermittently applied in an ON/OFF manner demonstrating temporal control over the nanoparticle morphology.

This article describes a process for producing polymeric self-assembled nanoparticles using visible light mediated dispersion polymerization. Using low energy visible light to control the polymerization allows for the reproducible formation of self-assembled worm-like micelles at high solids content.

Photoredox 촉매를 사용하여 가시 광선 중재 분산 중합에 의해 웜 형상 미셀 손쉬운 합성

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins ...

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Solid-phase synthesis has been used to obtain canonical and modified polymers of nucleic acids, specifically of DNA or RNA, which has made it a popular methodology for applications in various fields and for different research purposes. The procedure described herein focuses on the synthesis, purification, and characterization of dodecamers of RNA 5&#39;-[CUA CGG AAU CAU]-3&#39; containing zero, one, or two modifications located at the C2&#39;-O-position. The probes are based on 2-thiophenylmethyl groups, incorporated into RNA nucleotides via standard organic synthesis and introduced into the corresponding oligonucleotides via their respective phosphoramidites. This report makes use of phosphoramidite chemistry via the four canonical nucleobases (Uridine (U), Cytosine (C), Guanosine (G), Adenosine (A)), as well as 2-thiophenylmethyl functionalized nucleotides modified at the 2&#39;-O-position; however, the methodology is amenable for a large variety of modifications that have been developed over the years. The oligonucleotides were synthesized on a controlled-pore glass (CPG) support followed by cleavage from the resin and deprotection under standard conditions, i.e., a mixture of ammonia and methylamine (AMA) followed by hydrogen fluoride/triethylamine/N-methylpyrrolidinone. The corresponding oligonucleotides were purified via polyacrylamide electrophoresis (20% denaturing) followed by elution, desalting, and isolation via reversed-phase chromatography (Sep-pak, C18-column). Quantification and structural parameters were assessed via ultraviolet-visible (UV-vis) and circular dichroism (CD) photometric analysis, respectively. This report aims to serve as a resource and guide for beginner and expert researchers interested in embarking in this field. It is expected to serve as a work-in-progress as new technologies and methodologies are developed. The description of the methodologies and techniques within this document correspond to a DNA/RNA synthesizer (refurbished and purchased in 2013) that uses phosphoramidite chemistry.

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2&#039;-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

This article provides a detailed procedure on the solid-phase synthesis, purification, and characterization of dodecamers of RNA modified at the C2&#39;-O-position. UV-vis and circular dichroism photometric analyses are used to quantify and characterize structural aspects, i.e., single-strands or double-strands.

2&#39;- 포스 핀 올리고 뉴클레오타이드를 함유하는 올리고머의 고상 합성을위한 프로토콜<em&gt; O</em&gt; 원형 이색 성을 통한 - 티오 페닐 메틸 변형 및 특성 분석

Solid-phase synthesis has been used to obtain canonical and modified polymers of nucleic acids, specifically of DNA or RNA, which has made it a popular ...

Synthesis and Mass Spectrometry Analysis of Oligo-peptoids

Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have been thought of as a type of molecular information storage. Mass spectrometry analysis has been considered the method of choice for sequencing peptoids. Peptoids can be synthesized via solid phase chemistry using a repeating two-step reaction cycle. Here we present a method to manually synthesize oligo-peptoids and to analyze the sequence of the peptoids using tandem mass spectrometry (MS/MS) techniques. The sample peptoid is a nonamer consisting of alternating N-(2-methyloxyethyl)glycine (Nme) and N-(2-phenylethyl)glycine (Npe), as well as an N-(2-aminoethyl)glycine (Nae) at the N-terminus. The sequence formula of the peptoid is Ac-Nae-(Npe-Nme)4-NH2, where Ac is the acetyl group. The synthesis takes place in a commercially available solid-phase reaction vessel. The rink amide resin is used as the solid support to yield the peptoid with an amide group at the C-terminus. The resulting peptoid product is subjected to sequence analysis using a triple-quadrupole mass spectrometer coupled to an electrospray ionization source. The MS/MS measurement produces a spectrum of fragment ions resulting from the dissociation of charged peptoid. The fragment ions are sorted out based on the values of their mass-to-charge ratio (m/z). The m/z values of the fragment ions are compared against the nominal masses of theoretically predicted fragment ions, according to the scheme of peptoid fragmentation. The analysis generates a fragmentation pattern of the charged peptoid. The fragmentation pattern is correlated to the monomer sequence of the neutral peptoid. In this regard, MS analysis reads out the sequence information of the peptoids.

A protocol is described for the manual synthesis of oligo-peptoids followed by sequence analysis by mass spectrometry.

합성 및 올리고 peptoids의 질량 분석 분석

Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have ...