TW202005977A - 阿伐-選擇性唾液酸供體及其於製備唾液酸苷的用途 - Google Patents
阿伐-選擇性唾液酸供體及其於製備唾液酸苷的用途 Download PDFInfo
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Abstract
本揭示內容是關於一種唾液酸供體及其於合成神經節苷脂之用途。該唾液酸供體具有式(I)之結構,
其中R1及R2分別為可以一鹵素任選取代的苯甲醯基、甲苯磺醯基、三甲基乙醯基或乙醯基;且R3是乙醯基或-(O)CCH2OH。在一較佳的實施方式中,於式(I)之唾液酸供體中,R是乙醯基。本揭示內容亦提供一種用以合成一唾液酸苷的方法。該方法包含:在具有N-碘琥珀醯亞胺及三氟甲磺酸之適合條件下,將式(I)之唾液酸供體與一具有一級羥基之醣苷基受體耦合;以及單離該具有一α-醣苷鍵的唾液酸苷。依據較佳的實施方式,是於介於-20℃到-60℃的溫度中,在一選自CH3CN、CH3Cl及CH2Cl2所組成之群組的溶劑中進行耦合。此外或非必要性地,可於-40℃的溫度,在CH2Cl2中進行耦合。
Description
本揭示內容是關於合成唾液酸苷(sialoside)。具體來說,本揭示內容是關於一種新穎之唾液酸供體,其於合成一天然唾液酸苷時,可輔助進行α-立體選擇性(α-stereoselective)唾液酸化反應;以及該新穎之唾液酸供體於合成唾液酸苷的用途。
唾液酸是具有9個碳原子骨架之羧化糖的多樣性家族,廣泛表現於所有動物之細胞表面,且通常位於細胞表面之複合醣體的末端位置。其於諸如細胞-細胞附著及辨識、細胞分化、訊息傳遞及腫瘤轉移等不同生物及病理反應中扮演著功能性角色。因此,唾液酸醣苷及其寡聚物具有應用於醫藥領域之潛力,而合成包含唾液酸之複合醣體在醫藥領域亦極為重要。
天然唾液酸苷具有α-變旋異構 (α-anomer)結構。基於四級變旋異構中心的C-1羧基,且缺乏C-3位置之立體控制基調控α-立體選擇性唾液酸化反應,目前仍缺乏高產率、且可有效進行α-立體選擇性醣化反應的唾液酸供體。許多研究人員嘗試研發有效的策略及/或方法來進行α-唾液酸化反應,然而仍未產生另人滿意的結果。
有鑑於此,相關領域亟需一種經改善之策略,在不影響產率的情況下,具立體選擇性地合成具有α-變旋異構結構的唾液酸苷。
發明內容旨在提供本揭示內容的簡化摘要,以使閱讀者對本揭示內容具備基本的理解。此發明內容並非本揭示內容的完整概述,且其用意並非在指出本發明實施例的重要/關鍵元件或界定本發明的範圍。
本揭示內容是關於一種新穎且獨特的唾液酸供體,其於合成一天然唾液酸苷時,可在C-3位置進行α-立體選擇性唾液酸化反應,據以使唾液酸供體進行高度α-立體選擇性醣化反應,並改善其產率。因此,本揭示內容亦提供一種利用該新穎之唾液酸供體來合成一唾液酸苷的方法。
本揭示內容的第一態樣旨在提供一種唾液酸供體,其具有式(I)之結構,
其中,R1及R2分別為可以一鹵素任選取代的苯甲醯基(benzoyl)、甲苯磺醯基(甲苯sulfonyl)、三甲基乙醯基(pivaloyl)或乙醯基(acetyl);以及R3是乙醯基或-(O)CCH2OH。
依據一較佳實施方式,在式(I)之唾液酸供體中,R1及R2分別是苯甲醯基,且R3是乙醯基。
依據另一較佳實施方式,在式(I)之唾液酸供體中,R1及R2分別是苯甲醯基,且R3是-(O)CCH2OH。
本揭示內容的第二態樣是關於一種用以合成一唾液酸苷的方法。該方法包含:(a)在具有N-碘琥珀醯亞胺(N-iodosuccinimido,NIS)及三氟甲磺酸(triflueromothanesulfonic acid,TfOH)之適合條件下,將一唾液酸供體與一具有一級羥基之醣苷基受體耦合(coupling);以及(b)單離該唾液酸苷,其具有一α-醣苷鍵(α-glycosidic linkage);其中,該唾液酸供體具有式(I)之結構,
其中R1及R2分別為可以一鹵素任選取代的苯甲醯基、甲苯磺醯基、三甲基乙醯基或乙醯基;且R3是乙醯基或-(O)CCH2OH。
依據本揭示內容某些實施方式,步驟(a)是於介於-20℃到-60℃的溫度下,在一選自CH3CN、CH3Cl及CH2Cl2所組成之群組的溶劑中進行耦合。較佳地,是於-40℃的溫度,在CH2Cl2中進行耦合。
依據本揭示內容非必要性的實施方式,本發明方法步驟(a)是以一粉末分子篩來進行該耦合反應。
在參閱下文實施方式後,本發明所屬技術領域中具有通常知識者當可輕易瞭解本發明之基本精神及其他發明目的,以及本發明所採用之技術手段與實施態樣。
為了使本揭示內容的敘述更加詳盡與完備,下文針對了本發明的實施態樣與具體實施例提出了說明性的描述;但這並非實施或運用本發明具體實施例的唯一形式。實施方式中涵蓋了多個具體實施例的特徵以及用以建構與操作這些具體實施例的方法步驟與其順序。然而,亦可利用其他具體實施例來達成相同或均等的功能與步驟順序。
1.定義
以下闡述本揭示內容所使用的某些詞彙。除非本說明書另有定義,此處所用的科學與技術詞彙之含義與本發明所屬技術領域中具有通常知識者所理解與慣用的意義相同。
在本揭示內容中,「Pic」一詞是指皮考啉基(picolinyl)或2-吡啶甲基(2-pyridylmethyl);而「Pico」一詞則是指甲基砒碇(picoloyl)或2-吡啶羰基(2-pyridine carbonyl)。
「鹵素」(halo或halogen)一詞是指氟(fluorine,fluoro,-F)、氯(chlorine,chloro,-Cl)、溴(bromine,bromo,-Br)或碘(iodine,iodo,-I)。
除非另有所指,否則各苯甲醯基、甲苯磺醯基、三甲基乙醯基或乙醯基分別為可任選取代的(optionally substituted),亦即,可為未經取代的(一「未經取代的苯甲醯基」),或經一或多取代基取代的(一「經取代的乙醯基」)。當用以闡述一化學結構或化學域(moiety)時,「取代的」(substituted)一詞是指該結構或域的衍生物,其中是以一或多鹵素或羥基取代其氫原子。
若化合物具有一或多掌性中心,而其名稱未明確指出該些中心的立體化學式,則該名稱包含純立體異構物及其混合物。此外,應將圖式中非飽和價數之原子視為與足夠氫原子連接,以滿足其價數。
雖然用以界定本發明較廣範圍的數值範圍與參數皆是約略的數值,此處已盡可能精確地呈現具體實 施例中的相關數值。然而,任何數值本質上不可避免地含有因個別測試方法所致的標準偏差。在此處,「約」通常係指實際數值在一特定數值或範圍的正負10%、5%、1%或0.5%之內。或者是,「約」一詞代表實際數值落在平均值的可接受標準誤差之內,視本發明所屬技術領域中具有通常知識者的考量而定。除了實驗例之外,或除非另有明確的說明,當可理解此處所用的所有範圍、數量、數值與百分比(例如用以描述材料用量、時間長短、溫度、操作條件、數量比例及其他相似者)均經過「約」的修飾。因此,除非另有相反的說明,本說明書與附隨申請專利範圍所揭示的數值參數皆為約略的數值,且可視需求而更動。至少應將這些數值參數理解為所指出的有效位數與套用一般進位法所得到的數值。在此處,將數值範圍表示成由一端點至另一段點或介於二端點之間;除非另有說明,此處所述的數值範圍皆包含端點。
除非另有所指,否則本說明書所用的單數名詞涵蓋該名詞的複數型;而所用的複數名詞時亦涵蓋該名詞的單數型。
2.較佳實施方式說明
本揭示內容至少部分是基於發明人發現一種新穎之唾液酸供體,其可使一醣苷基受體(glycosyl acceptor)形成一α-定向(α-orientation)。非預期地,於-20℃到-60℃溫度時,在具有N-碘琥珀醯亞胺及催化性三氟甲磺酸之適當溶劑(例如CH2Cl2等)中,該新穎之唾液酸供體可高度α-立體選擇性(98-100%)地進行 醣化反應。可在短時間(例如,0.5小時)內完成該醣化反應,據以製備主要為α-變旋異構結構的產物。因此,本發明新穎之唾液酸供體可用以立體選擇性地合成具有α-變旋異構結構的天然唾液酸苷,其包含,但不限於,神經節苷脂(ganglioside)Hp-s1.
為確認較佳之可使醣苷基受體形成α-定向的唾液酸供體,本揭示內容合成數種具潛力之唾液酸供體,並分別與糖基受體反應。因此,本揭示內容的第一態樣是關於一種新穎之唾液酸供體,其具有式(I)之結構,
其中,R1及R2分別為可以一鹵素任選取代的苯甲醯基、甲苯磺醯基、三甲基乙醯基或乙醯基;且R3是乙醯基或-(O)CCH2OH。
在一較佳的實施方式中,式(I)之R1及R2分別是苯甲醯基,且R3是乙醯基,其為一種唾液酸供體(即,化合物1d),在與一糖基受體反應後,僅會產生α-變旋異構物(即,α-立體選擇性為100%)。
在另一較佳的實施方式中,式(I)之R1及R2分別是苯甲醯基,且R3是-(O)CCH2OH(即,化合物1h)。
本揭示內容的第二態樣旨在提供一種用以合成一唾液酸苷的方法。該方法包含:(a)在具有N-碘琥珀醯亞胺(NIS)及三氟甲磺酸(TfOH)之適合條件下,將一唾液酸供體與一具有一級羥基之醣苷基受體耦合;以及(b)單離該具有一α-醣苷鍵之唾液酸苷;其中,該唾液酸供體具有式(I)之結構,
其中,R1及R2分別為可以一鹵素任選取代的苯甲醯基、甲苯磺醯基、三甲基乙醯基或乙醯基;且R3是乙醯基或-(O)CCH2OH。
依據本揭示內容某些實施方式,在本發明方法之步驟(a)中,是於介於-20℃到-60℃的溫度下,在一選自CH3CN、CH3Cl及CH2Cl2所組成之群組的溶劑中進行耦合。
在本揭示內容一實施方式中,步驟(a)是於-20℃的溫度,在包含NIS及TfOH之CH2Cl2中進行耦合。該反應可於0.5小時內完成,具有中等α-立體選擇性,其中雙醣α-變旋異構物的產率約為β-變旋異構物之產率(約為24%)的5倍。
在本揭示內容另一實施方式中,步驟(a)是於-40℃的溫度,在包含NIS及TfOH之CH2Cl2中進行耦合。該反應可於2小時內完成,具有優異的α-立體選擇性,其中僅會產生α-變旋異構物,且產率約為70%。
在本揭示內容另一實施方式中,步驟(a)是於-60℃的溫度,在包含NIS及TfOH之CH2Cl2中進行耦合。該反應可於5小時內完成,具有優異的α-立體選擇性,其中僅會產生α-變旋異構物,且產率約為69%。
在本揭示內容再另一實施方式中,步驟(a)是於-40℃的溫度,在包含NIS及TfOH之CH3CN中進行耦合。該反應可於0.5小時內完成,具有優異的α-立體選擇性,其中僅會產生α-變旋異構物,且產率約為56%。
在本揭示內容進一步實施方式中,步驟(a)是於-40℃的溫度,在包含NIS及TfOH之CHCl3中進行耦合。該反應可於0.5小時內完成,具有優異的α-立體選擇性,其中僅會產生α-變旋異構物,惟產率僅約為20%。
非必要性地或此外,可加入粉末狀分子篩(例如,MS-3A)來進行本發明步驟(a)之耦合反應。
較佳地,步驟(a)製得的雙醣具有一α-變旋異構結構,且於步驟(b)可以任一種適合的方法(例如,管柱色層分析法)單離該雙醣。
依據本揭示內容某些實施方式,為確認最適合與本揭示內容之唾液酸供體耦合的醣苷基受體,本揭示內容合成數種具潛力的糖基受體,並於適當條件下,分別與本揭示內容之唾液酸供體反應;舉例來說,於-40℃,包含NIS及TfOH之CH2Cl2,並非必要性地可加入MS-3A。結果指出,為增加α-立體選擇性,唾液酸供體及糖基受體之間的耦合較佳是發生於一級羥基(或一級醇);因此,醣苷基受體的結構較佳包含一級醇。在某些實施例中,是利用一具有二級羥基之糖醇(例如,化合物5e)作為糖基受體,然而於本揭示內容之唾液酸供體及該 糖醇之間並不會發生耦合反應。
本揭示內容的另一態樣是關於一種用以合成一包含一α-變旋異構結構之天然神經節苷脂的方法。該方法是以式(I)之唾液酸供體作為起始材料,使一醣苷基受體形成α-定向。一般來說,依據欲合成之神經節苷脂的不同,式(I)之唾液酸供體可在本揭示內容確認之較佳的條件下(即,包含NIS及TfOH之CH2Cl2,反應溫度為-40℃,且非必要性地可加入MS-3A),與具有一級羥基之醣苷基受體反應,藉以製備一具有α-醣苷鍵的雙醣,其可以任何習知合成流程來製備特定的神經節苷脂。
依據本揭示內容之一較佳的實施方式,式(I)之唾液酸供體是作為用以製備Hp-s1的起始材料。較佳地,式(I)之唾液酸供體是先在-40℃溫度下,於包含NIS、TfOH及MS-3A之CH2Cl2中與S-噻唑基(S-thiazolyl)受體反應。反應會產生一僅具α-變旋異構結構之雙醣,其為去保護基之雙醣(即,移除雙醣之甲基砒碇),之後進一步與乙醯植物鞘胺醇(phytoceramide)反應,以得到具保護基之Hp-s1。接著將具保護基之Hp-s1進行一系列去保護基反應,包含去亞異丙基化(deisopropylidenenation)、去苄基化(debenzylation)、去乙醯基化(deacetylation)及皂化(saponification),據以製備特定的神經節苷脂Hp-s1。
下文提出多個實驗例來說明本發明的某些 態樣,以利本發明所屬技術領域中具有通常知識者實作本發明,且不應將這些實驗例視為對本發明範圍的限制。據信習知技藝者在閱讀了此處提出的說明後,可在不需過度解讀的情形下,完整利用並實踐本發明。此處所引用的所有公開文獻,其全文皆視為本說明書的一部分。
材料及方法
細胞株及細胞培養
由美國菌種保存中心(American Type Culture Collection,ATCC;Manassas,VA,USA)取得A549細胞。將A549細胞培養於包含每毫升1.5公克碳酸氫鈉、10%胎牛血清(fetal bovine serum,FBS)、1.0%抗生素(抗黴劑)、L-麩胺酸(2.0×10-3M)及1.0%非必需胺基酸之DMEM(Dulbecco’s minimal essential medium)細胞培養液中。分別利用台酚藍排除法(trypan blue exclusion method)及阿爾瑪藍法(Alamar Blue assay)來分析細胞數量及存活率。
實施例1 篩選唾液酸供體及評估唾液酸供體醣化的最佳條件
為確認適於使醣苷基受體形成α-定向之唾液酸供體,本實驗合成數種不同之唾液酸供體(詳見實施例1.1),並於適當環境下,與一葡苷基受體(glucosyl acceptor)耦合,之後單離並分析產物的α-立體選擇性(詳見實施例1.2)。確認化合物1d及1h為具有較高產 率之較佳唾液酸供體。
1.1 製備唾液酸供體
1.1.1 製備唾液酸供體1c
(苯基5-乙醯胺基-3,5-二脫氧-8,9-O-亞異丙基-2-硫基-D-丙三基-D-半乳-2-吡喃壬酮)甲酯(Methyl(phenyl 5-acetamido-3,5-dideoxy-8,9-O-isopropylidene-2-thio-D-glycero-D-galacto-2-nonulopyranosid)onate)(s2)
於0℃,將2,2-二甲氧基丙烷(4.4毫升,36.15毫莫耳)及CSA(4.454公克,19.28毫莫耳)加至包含化合物s1(10.006公克,24.10毫莫耳)之無水乙腈(240毫升)中。將混合物加熱至室溫,並於該溫度持續攪拌2小時。將Et3N(5.0毫升)加入反應混合物後,進行真空濃縮。利用MeOH及CH2Cl2(1:15, 體積比)作為沖提液,以快速管柱層析於矽膠純化觀察到的黃色漿狀產物,得到10.109克之白色固體s2,產率92%:Rf=0.25(MeOH:CH2Cl2=1:15(體積比));FT-IR(neat)vmax 3299,3083,2993,2953,1737,1660,1557,1439,1377,1263,1202,1135,1065,901,756,696,666,609cm-1;1H NMR(400MHz,CD3OD)δ□7.55-7.53(m,2H),7.39-7.36(m,1H),7.32-7.28(m,2H),4.34(dd,J=10.5,0.9Hz,1H),4.16-4.08(m,2H),3.98-3.82(m,3H),3.56(dd,J=7.5,0.9Hz,1H),3.51(s,3H),2.69(dd,J=13.6,4.7Hz,1H),2.02(s,3H),1.95(dd,J=13.6,11.8Hz,1H),1.39(s,3H),1.38(s,3H),1.30(s,3H),1.21(s,3H);13C NMR(100MHz,CD3OD)δ 173.4(C),169.2(C),136.0(CH),129.8(C),129.3(CH),128.5(CH),108.8(C),90.0(C),74.7(CH),72.6(CH),70.0(CH),66.6(CH2),66.4(CH),52.7(CH),51.8(CH3),48.5(CH3),40.9(CH2),25.9(CH3),24.7(CH3),21.6(CH3);HRMS-ESI[M+Na]+ Calcd for C21H29NO8SNa 478.1506,Found 478.1499.
(苯基5-乙醯胺基-3,5-二脫氧-8,9-O-亞異丙基-4,7-二-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-2-吡喃壬酮)甲酯(Methyl(phenyl 5-acetamido-3,5-dideoxy-8,9
-O-isopropylidene-4,7-di-O-picoloyl-2-thio-D-glycero-D-galacto-2-nonulopyranosid)onate)(s3)
於0℃,將2-吡啶甲酸(6.765公克,55.00毫莫耳)、DMAP(0.268公克,2.20毫莫耳)及DCC(10毫升,66.00毫莫耳)加至包含化合物s2(10.003公克,22.00毫莫耳)之無水二氯乙烷(73毫升)中。將混合物加熱至室溫,並於該溫度持續攪拌2小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用丙酮及乙酸乙酯(1:1,體積比)作為沖提液,以快速管柱層析於矽膠純化觀察到的黑色固體,得到13.919克之白色固體s3,產率92%:Rf=0.23(丙酮:乙酸乙酯=1:1(體積比));FT-IR(neat)vmax 3330,2988,1739,1675,1581,1543,1438,1373,1307,1245,1213,1131,1089,1053,994,751,701cm-1;1H NMR(400MHz,CDCl3)δ8.77(d,J=4.0Hz,1H),8.73(d,J=4.0Hz,1H),8.10(d,J=7.8Hz,1H),8.03(d,J=7.8Hz,1H),7.84(td,J=7.8,1.7Hz,1H),7.79(td,J=7.8,1.7Hz,1H),7.59-7.57(m,2H),7.49(m,5H),6.08(ddd,J=11.6,10.8,4.9Hz,1H),5.94-5.88(m,1H),5.81-5.80(m,1H),5.11(dd,J=10.5,1.8Hz,1H),4.14-4.06(m,2H),3.98-3.91(m,1H),3.73(dd,J=7.2,5.9Hz,1H),3.65(s,3H),2.93(dd,J=13.7,4.9Hz,1H),2.26(dd,J=13.7,11.6Hz,1H),1.90(s,3H),1.29(s,3H),1.21(s,3H);13C NMR(75MHz,CDCl3)δ170.7(C),170.6(C),168.3(C),164.4(C),164.3(C),150.0(CH),149.9(CH),147.6(C),147.2(C),137.2(CH),137.1(CH),136.9(CH),136.3(CH),129.8(CH),129.3(CH), 128.9(CH),127.2(CH),127.0(CH),125.6(CH),125.5(CH),108.5(C),108.4(C),88.8(C),87.7(C),75.4(CH),71.7(CH),71.0(CH),70.7(CH),70.2(CH),65.8(CH2),65.5(CH2),52.9(CH3),52.5(CH3),50.6(CH),38.1(CH2),26.3(CH3),26.2(CH3),25.4(CH3),25.1(CH3),23.3(CH3);HRMS-ESI[M+Na]+ Calcd for C33H35N3O11SNa 688.1935,Found 688.1918.
(苯基5-乙醯胺基-3,5-二脫氧-4,9-二-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-3,5-dideoxy-4,9-di-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(s4)
於0℃,將異亞丙基縮醛s3(8.000公克,12.79毫莫耳)溶於80%AcOH溶液(120毫升)中,於70℃持續攪拌混合物4小時。將反應混合物與甲苯進行共蒸發,以得到一白色固體殘留物。使6.467克之s4再結晶(MeOH/CH2Cl2)後,得到一白色固體化合物,產率86%:Rf=0.28(MeOH:CH2Cl2=1:9(體積比));FT-IR(neat)vmax 3311,3064,3009,2954,1732,1659,1555,1439,1375,1309,1246,1136,1089,1047,998,752,701cm-1;1H NMR(400MHz,CD3OD)δ 8.69-8.68(m,2H),8.22(dt,J=7.9,0.9Hz,1H),8.09(dt,J=7.9,0.9Hz,1H), 8.00(td,J=7.8,1.6Hz,1H),7.91(td,J=7.8,1.6Hz,1H),7.65-7.54(m,4H),7.36-7.31(m,3H),5.68(ddd,J=11.6,8.4,4.8Hz,1H),4.75-4.69(m,2H),4.43-4.36(m,2H),4.16(ddd,J=9.0,6.6,2.3Hz,1H),3.7(dd,J=9.0,0.8Hz,1H),3.50(s,3H),2.94(dd,J=13.5,4.8Hz,1H),2.28(dd,J=13.5,11.6Hz,1H),1.99(s,3H);13C NMR(75MHz,CD3OD)δ173.8(C),173.5(C),172.9(C),169.2(C),168.3(C),164.8(C),164.7(C),164.6(C),150.2(CH),149.7(CH),149.2(CH),147.8(C),147.6(C),146.9(C),137.7(CH),137.5(CH),137.3(CH),136.9(CH),136.1(CH),129.5(CH),129.5(CH),128.9(CH),128.8(CH),127.5(CH),127.3(CH),125.6(CH),125.3(CH),89.6(C),72.5(CH),70.5(CH),69.4(CH),68.7(CH2),68.3(CH),52.5(CH3),50.8(CH3),38.0(CH2),23.0(CH3),22.9(CH3);HRMS-ESI[M+Na]+ Calcd for C30H31N3O10SNa 648.1622,Found 648.1629.
(苯基5-乙醯胺基-7,8-二-O-苯甲醯基-3,5-二脫氧-4,9-二-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-7,8-di-O-benzoyl-3,5-dideoxy-4,9-di-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranosi
de)onate)(1c)
於0℃,將苯甲醯氯(0.08毫升,0.70毫莫耳)加至包含化合物s4(0.200公克,0.32毫莫耳)之無水吡啶(1.6毫升)中。於該溫度持續攪拌混合物5小時。將反應混合物置於冰上後,以乙酸乙酯萃取,並利用飽和K2CO3溶液及鹽水洗滌產物,之後以MgSO4進行乾燥處理及真空濃縮。利用乙酸乙酯及正己烷(2.5:1,體積比)作為沖提液,以快速管柱層析於矽膠純化得到的白色固體殘留物,以製備0.255克之1c白色固體化合物,產率96%:Rf=0.28(乙酸乙酯:正己烷=3:1(體積比));FT-IR(neat)vmax 3324,3066,2919,2852,1731,1674,1546,1443,1369,1266,1097,1035,894,754,712,610cm-1;1H NMR(400MHz,CDCl3)δ 8.92(d,J=4.1Hz,1H),8.77(d,J=4.1Hz,1H),8.07-7.97(m,6H),7.85-7.78(m,2H),7.60(t,J=7.4Hz,1H),7.54-7.44(m,5H),7.40-7.36(m,4H),7.14-7.09(m,3H),6.97(d,J=9.6Hz,1H),6.06(dd,J=4.2,1.8Hz,1H),5,91(ddd,J=13.5,10.1,4.9Hz,1H),5.71(ddd,J=6.5,4.8,1.8Hz,1H),5.12(dd,J=10.1,4.2Hz,1H),5.07(dd,J=11.8,4.8Hz,1H),4.58(dd,J=11.8,7.1Hz,1H),4.38(q,J=10.1Hz,1H),3.54(s,3H),2.84(dd,J=13.5,4.9Hz,1H),2.35(dd,J=13.5,11.8Hz,1H),1.85(s,3H);13C NMR(100MHz,CDCl3)δ170.4(C),168.0(C),165.8(C),164.4(C),164.4(C),163.8(C),150.0(CH),149.9(CH),147.3(C),147.0(C),137.3(CH),136.9(CH),136.1(CH),133.1(CH),133.0(CH),129.8(CH),129.7(CH),129.6(C),129.4(CH),129.3(C),129.0(C),128.8(CH),128.3(CH), 128.3(CH),127.1(CH),126.8(CH),125.7(CH),125.0(CH),88.9(C),73.0(CH),72.7(CH),71.0(CH),70.8(CH),63.8(CH2),52.3(CH3),49.5(CH),37.5(CH2),23.0(CH3);HRMS-ESI[M+Na]+ Calcd for C44H39N3O12SNa 856.2147,Found 856.2146.
1.1.2 製備唾液酸供體1a及1e
(苯基5-乙醯胺基-4,8,9-三-O-苯甲醯基-3,5-二脫氧-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-4,8,9-tri-O-benzoyl-3,5-dideoxy-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate(s5);以及(苯基5-乙醯胺基-4,7,8,9-四-O-苯甲醯基-3,5-二脫氧-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯Methyl(phenyl 5-acetamido-4,7,8,9-tetra-O-benzoyl-3,5-dideoxy-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1e)
於0℃,將苯甲酐(1.085公克,4.80毫莫耳)加至包含化合物s1(0.503公克,1.20毫莫耳)、Im(0.410公克,6.02毫莫耳)、DMAP(0.073公克,0.60毫莫耳)之無水二氯乙烷(6.0毫升)中。將混合物加熱至60℃,並於該溫度持續攪拌11小時。冰浴反應混合物並加入6N HCl後,以乙酸乙酯萃取,之後利用飽和K2CO3溶液及鹽水洗滌產物,以MgSO4進行乾燥處理及真空濃縮。利用乙酸乙酯及正己烷(1:2,體積比)作為沖提液,以快速管柱層析於矽膠純化得到的白色固體殘留物,以製備0.368克之s5白色固體化合物,產率42%,以及0.431克之白色固體化合物1e,產率43%:s5:Rf=0.38(乙酸乙酯:正己烷=1:1(體積比));FT-IR(neat)vmax 3355,3066,2954,1721,1663,1602,1548,1448,1373,1269,1172,1112,1070,1027,755,712cm-1;1H NMR(400MHz,CDCl3)δ 8.11-8.08(m,2H),8.02-8.00(m,2H),7.92-7.89(m,2H),7.62-7.37(m,11H),7.20-7.16(m,2H),7.06-7.02(m,1H),6.41(d,J=8.0Hz,1H),5.76(ddd,J=11.8,10.3,4.8Hz,1H),5.48-5.45(m,1H),4.95(dd,J=12.2,2.2Hz,1H),4.92(d,J=4.1Hz,1H),4.66(dd,J=12.2,8.2Hz,1H),4.48(dd,J=10.3,1.4Hz,1H),4.24-4.17(m,1H),4.10(bs,1H),3.55(s,3H),2.87(dd,J=13.8,4.8Hz,1H),2.41(dd,J=13.8,11.8Hz,1H),1.96(s,3H);13C NMR(75MHz,CDCl3)δ 173.2(C),168.6(C),167.1(C),166.4(C),165.9(C),136.1(CH),133.8(CH),133.2(CH),132.8(CH),130.3(CH),130.0(CH),129.8(CH),129.7(CH),129.6(CH),129.0(CH),128.9(CH),128.6(CH),128.6(CH),128.4(CH),128.3(CH),89.3(C),74.8(CH), 74.4(CH),69.9(CH),69.2(CH),64.0(CH2),52.6(CH3),51.8(CH),37.8(CH2),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C39H37NO11SNa 750.1980,Found 750.1969;1e:Rf=0.50(乙酸乙酯:正己烷=1:1(體積比));FT-IR(neat)vmax 3372,3066,1724,1601,1536,1449,1369,1265,1175,1105,1070,1026,892,755,712cm-1;1H NMR(400MHz,CDCl3)δ 8.12-7.93(m,9H),7.65-7.31(m,15H),7.24-7.22(m,1H),7.11-7.07(m,1H),6.03-6.01(m,1H),5.86-5.86(m,1H),5.76-5.69(m,2H),5.56-5.53(m,2H),5.29-5.22(m,1H),5.03-4.94(m,3H),4.68-4.64(m,1H),4.52(dd,J=12.2,8.8Hz,1H),4.37(q,J=10.5Hz,1H),4.28(d,J=10.5Hz,1H),4.14(q,J=9.9Hz,1H),3.65(s,3H),3.42(s,3H),3.06(dd,J=12.8,4.7Hz,1H),2.95(dd,J=13.8,4.7Hz,1H),2.32(dd,J=13.8,11.8Hz,1H),2.12(dd,J=12.8,11.8Hz,1H),1.81(s,3H),1.79(s,3H);13C NMR(75MHz,CDCl3)δ170.6(C),170.6(C),168.8(C),168.6(C),166.9(C),166.6(C),166.5(C),166.3(C),165.9(C),165.9(C),165.5(C),136.9(CH),136.2(CH),133.5(CH),133.5(CH),133.4(CH),133.4(CH),133.3(CH),133.1(CH),133.0(CH),130.1(CH),130.0(CH),129.9(CH),129.8(CH),129.8(CH),129.7(CH),129.7(CH),129.6(CH),129.3(CH),129.2(CH),129.0(CH),128.8(CH),128.6(CH),128.5(CH),128.4(CH),128.3(CH),89.2(C),88.0(C),75.6(CH),74.2(CH),73.5(CH),71.9(CH),70.8(CH),70.6(CH),70.4(CH),69.3(CH),63.6(CH2),63.2(CH2),52.8(CH3),52.7(CH3),49.6(CH),38.3 (CH2),38.0(CH2),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C46H41NO12SNa 854.2242,Found 854.2253.
(苯基5-乙醯胺基-4,8,9-三-O-苯甲醯基-3,5-二脫氧-7-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-4,8,9-tri-O-benzoyl-3,5-dideoxy-7-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1a)
於0℃,將2-吡啶甲酸(0.076公克,0.62毫莫耳)、DMAP(0.005公克,0.04毫莫耳)及DCC(0.130公克,0.82毫莫耳)加至包含化合物s5(0.305公克,0.41毫莫耳)之無水CH2Cl2(2.0毫升)中。將混合物加熱至室溫,並於該溫度持續攪拌1.5小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯及正己烷(1:1,體積比)作為沖提液,以快速管柱層析於矽膠純化觀察到的黑色固體,得到0.346g克之白色固體1a,產率99%:Rf=0.13(乙酸乙酯:正己烷=1:1(體積比));FT-IR(neat)vmax 3371,3064,2931,2853,1724,1548,1445,1367,1273,1111,1027,892,752,712cm-1;1H NMR(400MHz,CDCl3)δ 8.81-8.79(m,1H),8.79-8.77(m,1H),8.19-8.16(m,1H),8.16-8.13(m,1H),8.03-7.82(m,8H),7.63-7.61(m, 2H),7.55-7.44(m,7H),7.41-7.35(m,7H),7.27-7.24(m,4H),7.14-7.10(m,1H),6.06(t,J=2.2Hz,1H),5.94-5.88(m,2H),5.78-5.71(m,1H),5.70(d,J=9.9Hz,1H),5.61(dt,J=8.4,2.2Hz,1H),5.53(d,J=9.5Hz,1H),5.32-5.25(m,1H),5.07(dd,J=10.9,2.9Hz,1H),5.01(dd,J=12.2,2.2Hz,1H),4.95(dd,J=12.2,2.2Hz,1H),4.68(dd,J=12.3,5.8Hz,1H),4.54(dd,J=12.3,8.6Hz,1H),4.39(q,J=10.2Hz,1H),4.34(dd,J=10.2,2.2Hz,1H),4.14(q,J=10.2Hz,1H),3.62(s,3H),3.67(s,3H),3.09(dd,J=12.8,4.7Hz,1H),2.95(dd,J=13.8,4.7Hz,1H),2.34(dd,J=13.8,11.5Hz,1H),2.13(dd,J=12.8,11.5Hz,1H),1.80(s,3H),1.78(s,3H);13C NMR(75MHz,CDCl3)δ170.5(C),68.6(C),168.4(C),166.6(C),166.3(C),166.2(C),166.2(C),165.8(C),165.6(C),163.9(C),163.9(C),150.0(CH),147.4(C),147.3(C),137.1(CH),136.7(CH),136.2(CH),133.4(CH),133.3(CH),133.0(CH),132.9(CH),129.8(CH),129.7(CH),129.6(CH),129.5(CH),129.2(CH),129.1(CH),128.9(CH),128.8(CH),128.7(CH),128.5(CH),128.4(CH),128.3(CH),128.2(CH),127.1(CH),125.5(CH),89.1(C),88.1(C),75.3(CH),73.7(CH),73.3(CH),71.4(CH),70.2(CH),(CH),63.5(CH2),63.0(CH2),52.7(CH3),52.7(CH3),49.8(CH),48.9(CH),38,4(CH2),38.0(CH2),33.8(CH2),23.0(CH3);HRMS-ESI[M+Na]+ Calcd for C45H40N2O12SNa 855.2194,Found 855.2194.
1.1.3 製備唾液酸供體1d、1f及1g
(苯基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(s6)
於0℃,將苯甲酐(10.848公克,48.12毫莫耳)加至包含化合物s1(5.030公克,12.02毫莫耳)、Im(4.096公克,60.21毫莫耳)及DMAP(0.732公克,0.60毫莫耳)之無水二氯乙烷(120毫升)中。將混合物加熱至40℃,並於該溫度持續攪拌5小時。冰浴反應混合物並加入6N HCl,以乙酸乙酯萃取後,利用飽和K2CO3溶液及鹽水洗滌產物,之後以MgSO4進行乾燥處理及真空濃縮。利用乙酸乙酯及正己烷(1:1,體積比)作為沖提液,以快速管柱層析於矽膠純化得到的白色固體殘留物,得到5.263克之白色固體化合物s6,產率70%,以及1.147克之白色固體化合物s5,產率13%:Rf=0.13(乙酸乙酯:正己烷=1:1(體積比));FT-IR(neat)vmax 3349,3065,2953,1720,1660,1603,1551,1446,1375,1272,1116,1070,1027,754, 713cm-1;1H NMR(400MHz,CDCl3)δ 8.09-8.00(m,4H),7.61-7.53(m,3H),7.50-7.39(m,4H),7.32-7.28(m,4H),6.42(d,J=8.2Hz,1H),6.29(d,J=8.2Hz,1H),5.76-5.69(m,1H),5.27-5.20(m,1H),4.74(dd,J=11.4,2.4Hz,1H),4.63(dd,J=11.4,2.4Hz,1H),4.57(d,J=7.2Hz,1H),4.53-4.52(m,1H),4.50(dd,J=6.4,5.6Hz,1H),4.28-4.20(m,2H),4.18-4.13(m,2H),3.72(dd,J=8.7,0.8Hz,1H),3.61(d,J=8.7Hz,1H),3.53(s,3H),3.51(s,3H),3.00(dd,J=12.9,4.8Hz,1H),2.87(dd,J=13.7,4.8Hz,1H),2.36(dd,J=13.7,11.8Hz,1H),1.95(s,3H),1.92(s,3H);13C NMR(75MHz,CDCl3)δ173.2(C),172.8(C),169.5(C),168.4(C),167.2(C),167.1(C),166.9(C),137.0(CH),135.7(CH),133.8(CH),133.1(CH),130.1(CH),129.9(CH),129.7(CH),129.6(CH),129.6(CH),129.5(CH),129.0(CH),128.9(CH),128.8(CH),128.7(CH),128.6(CH),128.4(CH),128.3(CH),89.4(C),85.8(C),73.0(CH),69.5(CH),69.3(CH),69.2(CH),67.5(CH2),52.9(CH3),52.6(CH3),51.5(CH),51.0(CH),38.1(CH2),37.3(CH2),23.2(CH3),22.9(CH3);HRMS-ESI[M+Na]+ Calcd for C32H33NO10SNa 646.1717,Found 646.1708.
(苯基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧
-7,8-二-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯Methyl(phenyl(5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1d)
於0℃,將2-吡啶甲酸(1.867公克,15.21毫莫耳)、DMAP(0.074公克,0.61毫莫耳)及DCC(3.754公克,18.21毫莫耳)加至包含化合物s6(3.790公克,6.07毫莫耳)之無水二氯乙烷(30毫升)中。將混合物加熱至40℃,並於該溫度持續攪拌4小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯作為沖提液,以快速管柱層析於矽膠純化觀察到的黑色固體,以得到4.938克之白色固體1d,產率97%:Rf=0.30(乙酸乙酯);FT-IR(neat)vmax 3275,3063,1727,1680,1582,1550,1442,1368,1275,1119,1027,996,751,712cm-1;1H NMR(400MHz,CDCl3)δ 8.80-8.78(m,1H),8.77-8.74(m,2H),8.70-8.69(m,1H),8.18(d,J=7.8Hz,1H),8.14(d,J=7.8Hz,1H),8.10(d,J=7.8Hz,1H),8.05(d,J=7.8Hz,1H),7.97-7.76(m,8H),7.63-7.32(m,12H),7.13-7.09(m,1H),6.21(d,J=9.7Hz,1H),6.08(t,J=2.4Hz,1H),6.04(dd,J=6.4,1.3Hz,1H),5.99-5.95(m,2H),5.75-5.68(m,1H),5.64(dt,J=8.4,2.2Hz,1H),5.19-5.13(m,1H),5.06-5.02(m,2H),4.95(dd,J=12.3,2.4Hz,1H),4.66(dd,J=12.3,6.0Hz,1H),4.54(dd,J=12.3,8.4Hz,1H),4.46(q,J=10.1Hz,1H),4.33-4.31(m,1H), 4.25(q,J=10.1Hz,1H),3.64(s,3H),3.32(s,3H),3.07(dd,J=12.9,4.8Hz,1H),2.94(dd,J=13.9,4.8Hz,1H),2.32(dd,J=13.9,11.8Hz,1H),2.12(dd,J=12.9,11.8Hz,1H),1.80(s,3H),1.78(s,3H);13C NMR(75MHz,CDCl3)δ170.7(C),170.5(C),168.6(C),168.2(C),166.5(C),166.3(C),166.1(C),165.7(C),164.4(C),163.9(C),163.8(C),150.0(CH),149.9(CH),149.6(CH),147.5(C),147.1(C),137.5(CH),137.4(CH),137.0(CH),136.8(CH),136.1(CH),133.4(CH),132.9(CH),129.8(CH),129.8(CH),129.6(CH),129.5(CH),129.4(CH),129.1(CH),128.9(CH),128.6(CH),128.5(CH),128.2(CH),127.5(CH),127.3(CH),127.1(CH),126.9(CH),125.9(CH),125.6(CH),88.8(C),88.0(C),75.5(CH),74.8(CH),73.3(CH),72.7(CH),71.0(CH),70.6(CH),63.2(CH2),62.8(CH2),52.6(CH3),52.6(CH3),49.2(CH),49.0(CH),37.7(CH2),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C44H39N3O12SNa 856.2147,Found 856.2147.
(苯基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-7,8-二-O-菸鹼醯基-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-nicotinoyl-2-
thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1f)
於0℃,將菸鹼酸(0.246公克,2.00毫莫耳)、DMAP(0.010公克,0.08毫莫耳)及DCC(0.380公克,2.40毫莫耳)加至包含化合物s6(0.500公克,0.80毫莫耳)之無水二氯乙烷(4.0毫升)中。將混合物加熱至27℃,並於該溫度持續攪拌1小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯作為沖提液,以快速管柱層析於矽膠純化觀察到的黃色固體,以得到0.621克之白色固體1f,產率93%:Rf=0.38(乙酸乙酯);FT-IR(neat)vmax 3269,3062,1728,1684,1591,1543,1475,1446,1369,1272,1201,1109,1026,749,712cm-1;1H NMR(400MHz,CDCl3)δ 9.30(dd,J=2.1,0.7Hz,1H),9.28(dd,J=2.1,0.7Hz,1H),9.21(dd,J=2.1,0.7Hz,1H),9.18(dd,J=2.1,0.7Hz,1H),8.83-8.74(m,4H),8.33-8.29(m,2H),8.26-8.22(m,2H),7.96-7.91(m,6H),7.86-7.84(m,2H),7.63-7.34(m,22H),7.15-7.11(m,1H),5.94(t,J=2.0Hz,1H),5.88-5.84(m,1H),5.80(dd,J=5.5,0.7Hz,1H),5.74-5.67(m,1H),5.56(d,J=10.0Hz,1H),5.53(dt,J=8.5,2.0Hz,1H),5.37-5.34(m,1H),5.17-5.10(m,1H),4.96-4.88(m,3H),4.65(dd,J=12.4,6.3Hz,1H),4.50(dd,J=12.4,8.6Hz,1H),4.39(q,J=10.3Hz,1H),4.26-4.20(m,2H),3.69(s,3H),3.54(s,3H),3.04(dd,J=12.9,4.7Hz,1H),2.90(dd,J=13.9,4.9Hz,1H),2.35(dd,J=13.9,11.7Hz,1H),2.17(dd,J=12.9,11.7Hz,1H),1.79(s,3H),1.76(s,3H);13C NMR(100MHz, CDCl3)δ170.4(C),170.3(C),168.5(C),168.3(C),166.7(C),166.6(C),166.2(C),165.8(C),165.2(C),164.6(C),164.5(C),164.4(C),153.8(CH),151.3(CH),151.2(CH),151.2(CH),137.6(CH),137.5(CH),137.4(CH),137.4(CH),136.9(CH),136.2(CH),133.6(CH),133.6(CH),133.3(CH),133.1(CH),130.3(CH),130.0(CH),129.9(CH),129.9(CH),129.7(C),129.6(CH),129.6(C),129.5(CH),129.3(CH),129.1(CH),128.7(CH),128.7(CH),128.6(C),128.5(CH),128.5(CH),128.5(CH),128.4(C),126.0(C),125.7(C),125.7(C),123.6(CH),123.5(CH),89.0(C),87.7(C),75.8(CH),74.5(CH),73.7(CH),72.1(CH),70.7(CH),70.4(CH),69.9(CH),69.7(CH),63.0(CH2),62.7(CH2),53.0(CH3),52.9(CH3),49.9(CH),49.7(CH),38.5(CH2),38.0(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C44H39N3O12SNa 856.2147,Found 856.2132.
[苯基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-7,8-二-O-異菸鹼醯基-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷]甲酯(Methyl[phenyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-isonicotinoyl-2-thio-D-glycero-D-galacto-non-2-ulopyrano
side]onate)(1g)
於0℃,將異菸鹼酸(0.246公克,2.00毫莫耳)、DMAP(0.010公克,0.08毫莫耳)及DCC(0.380公克,2.40毫莫耳)加至包含化合物s6(0.501公克,0.80毫莫耳)之無水二氯乙烷(4.0毫升)中。將混合物加熱至40℃,並於該溫度持續攪拌2小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯作為沖提液,以快速管柱層析於矽膠純化觀察到的黃色固體,以得到0.601克之白色固體1g,產率90%:Rf=0.33(乙酸乙酯);FT-IR(neat)vmax 3374,3061,1728,1684,1601,1560,1446,1409,1369,1271,1211,1113,1067,1027,997,755,712cm-1;1H NMR(400MHz,CDCl3)δ□8.83-8.74(m,8H),7.95-7.91(m,6H),7.86-7.83(m,6H),7.79-7.78(m,4H),7.63-7.60(m,2H),7.56-7.52(m,6H),7.41-7.28(m,12H),7.18-7.14(m,1H),5.92(t,J=2.0Hz,1H),5.87-5.84(m,1H),5.77(dd,J=5.5,0.7Hz,1H),5.73-5.66(m,1H),5.54(d,J=10.0Hz,1H),5.49(dt,J=8.4,2.1Hz,1H),5.37(d,J=8.2Hz,1H),5.16-5.10(m,1H),4.95-4.88(m,3H),4.64(dd,J=12.6,6.3Hz,1H),4.52(dd,J=12.6,8.4Hz,1H),4.37(q,J=10.2Hz,1H),4.21-4.17(m,2H),3.70(s,3H),3.55(s,3H),3.05(dd,J=12.9,4.7Hz,1H),2,93(dd,J=13.9,4.8Hz,1H),2.34(dd,J=13.9,11.6Hz,1H),2.19(dd,J=12.9,12.0Hz,1H),1.79(s,3H),1.76(s,3H);13C NMR(100MHz,CDCl3)δ170.4(C),168.4(C),168.4(C),168.2(C),166.7(C),166.7(C),166.2(C),165.8(C),165.0(C),164.5(C),164.4(C),164.3(C),150.8(CH),150.7(CH), 150.5(CH),137.3(C),137.0(CH),136.9(C),136.9(C),136.1(CH),133.7(CH),133.4(CH),133.2(CH),130.3(CH),130.1(CH),129.9(CH),129.7(CH),129.6(CH),129.4(CH),129.1(CH),129.1(CH),128.7(CH),128.5(CH),128.4(CH),123.2(CH),123.1(CH),123.1(CH),89.0(C),88.9(C),87.7(C),75.7(CH),74.7(CH),73.7(CH),72.2(CH),71.0(CH),70.3(CH),69.9(CH),69.8(CH),62.8(CH2),62.5(CH2),52.9(CH3),49.9(CH),49.7(CH),38.6(CH2),38.0(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C44H39NO12SNa 856.2147,Found 856.2153.
1.1.4 製備唾液酸供體1d及1b
(苯基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-8-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖
苷)甲酯(Methyl(phenyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-8-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(s7)
於0℃,將2-吡啶甲酸(0.151公克,1.22毫莫耳)、DMAP(0.088公克,0.07毫莫耳)及EDC(0.276公克,1.44毫莫耳)加至包含化合物s6(0.450公克,0.72毫莫耳)之無水CH2Cl2(3.6毫升)中。將混合物加熱至18℃,並於該溫度持續攪拌20小時。以鹽水洗滌反應混合物,之後利用MgSO4進行乾燥處理,並真空濃縮。利用乙酸乙酯及正己烷(3:1,體積比),以快速管柱層析於矽膠純化觀察到的黑色固體殘留物,之後以乙酸乙酯進行沖提,得到0.113克之白色固體s6,產率25%,以及0.226克之白色固體化合物s7,產率43%,以及0.120克之白色固體化合物1d,產率20%:Rf=0.20(乙酸乙酯:正己烷=3:1(體積比));FT-IR(neat)vmax 3288,3065,1726,1662,1582,1539,1446,1375,1263,1176,1108,1069,1025,754,713cm-1;1H NMR(400MHz,CDCl3)δ□8.72(d,J=2.7Hz,1H),8.16(d,J=7.8Hz,1H),8.01(dd,J=8.4,1.3Hz,2H),7.90(dd,J=8.4,1.3Hz,2H),7.80(td,J=7.7,1.7Hz,1H),7.60(t,J=7.4Hz,1H),7.53-7.35(m,8H),7.18(t,J=7.6Hz,2H),7.02(t,J=7.4Hz,1H),6.85-6.80(m,1H),5.78-5.72(m,1H),5.54(dt,J=8.3,2.5Hz,1H),4.96(dd,J=12.3,2.0Hz,1H),4.55(d,J=10.2Hz,1H),4.28-4.18(m,2H),3.57(s,3H),2.85(dd,J=13.8,4.8 Hz,1H),2.35(dd,J=13.8,11.6Hz,1H),1.94(s,3H);13C NMR(75MHz,CDCl3)δ 173.3(C),168.7(C),167.0(C),165.9(C),164.6(C),150.1(CH),149.9(CH),149.6(CH),147.7(C),146.8(C),137.3(CH),137.2(CH),137.1(CH),136.5(CH),136.1(CH),133.9(CH),133.6(CH),133.5(CH),133.1(CH),132.9(CH),132.8(CH),130.3(C),129.9(CH),129.8(CH),129.6(CH),129.1(CH),128.9(CH),128.8(CH),128.7(CH),128.4(CH),127.3(CH),127.2(CH),127.0(CH),125.7(CH),125.6(CH),89.2(C),87.8(C),76.2(CH),75.4(CH),74.7(CH),74.4(CH),72.4(CH),72.1(CH),70.3(CH),70.0(CH),69.7(CH),69.3(CH),67.7(CH),65.2(CH),64.0(CH2),63.2(CH2),52.7(CH3),52.3(CH3),51.7(CH),51.5(CH),37.8(CH2),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C38H36N2O11SNa 751.1932,Found 751.1937.
(苯基5-乙醯胺基-4,7,9-三-O-苯甲醯基-3,5-二脫氧-8-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 5-acetamido-4,7,9-tri-O-benzoyl-3,5-dideoxy-8-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1b)
於0℃,將苯甲酸(0.038公克,0.32毫莫耳)、DMAP(0.003公克,20.58微莫耳)及DCC(0.085公克,0.42毫莫耳)加至包含化合物s7(0.150公克,0.21毫莫耳)之無水CH2Cl2(2.0毫升)中。將混合物加熱至40℃,並於該溫度持續攪拌2小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯及CH2Cl2(1:2,體積比)作為沖提液,以快速管柱層析於矽膠純化觀察到的白色固體,以得到0.145克之白色固體1b,產率83%:Rf=0.45(乙酸乙酯:CH2Cl2=1:1(體積比));FT-IR(neat)vmax 3311,3066,3011,2922,2852,1729,1682,1547,1444,1368,1272,1109,1032,892,755,713,609cm-1;1H NMR(400MHz,CDCl3)δ 8.80(d,J=4.8Hz,1H),8.77(d,J=4.1Hz,1H),8.18(d,J=7.5Hz,1H),8.16(d,J=8.1Hz,1H),8.07-7.93(m,10H),7.88-7.86(m,3H),7.79(td,J=7.8,1.5Hz,1H),7.63-7.23(m,19H),7.11(t,J=7.4Hz,1H),6.29(d,J=8.7Hz,1H),6.08-6.07(m,1H),6.02-6.01(m,1H),5.96-5.90(m,2H),5.79-5.68(m,2H),5.60-5.58(m,1H),5.30-5.24(m,1H),5.02-4.95(m,1H),4.68(dd,J=12.3,6.2Hz,1H),4.52(dd,J=12.3,8.7Hz,1H),4.42(q,J=10.2Hz,1H),4.34-4.32(m,1H),4.16(q,J=10.6Hz,1H),3.63(s,3H),3.36(s,3H),3.09(dd,J=12.5,4.6Hz,1H),2.94(dd,J=13.8,4.5Hz,1H),2.31(dd,J=13.8,11.6Hz,1H),2.13(dd,J=12.5,11.5Hz,1H),1.82(s,3H),1.79(s,3H);13C NMR(100MHz,CDCl3)δ170.5(C),168.6(C),166.6(C),165.8(C),165.5(C),164.4(C),150.0(CH),147.2(C),137.5(CH),136.0(CH),133.4(CH),133.2 (CH),132.9(CH),129.9(CH),129.9(CH),129.8(C),129.7(CH),129.5(CH),129.4(CH),129.0(CH),128.7(C),128.6(CH),128.5(CH),128.3(CH),128.2(CH),127.5(CH),125.8(CH),88.6(C),75.1(CH),73.2(CH),70.8(CH),70.0(CH),63.2(CH2),52.7(CH3),49.2(CH),37.7(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C45H40N2O12SNa 855.2194,Found 855.2181.
1.1.5 製備唾液酸供體1h
(苯基4,9-二-O-苯甲醯基-5-苄氧乙醯胺基-3,5-二脫氧-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 4,9-di-O-benzoyl-5-benzyloxyacetamido-3,5-dideoxy-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(s9);以及(苯基4,8,9-三-O-苯甲醯基-5-苄氧乙醯胺基-3,5-二脫氧-8-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯
(Methyl(phenyl 4,8,9-tri-O-benzoyl-5-benzyloxyacetamido-3,5-dideoxy-8-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(s10)
於0℃,將苯甲酐(3.637公克,16.08毫莫耳)加至包含化合物s8(2.100公克,4.02毫莫耳)、Im(1.368公克,20.11毫莫耳)及DMAP(0.245公克,2.01毫莫耳)之無水二氯乙烷(40毫升)中。將混合物加熱至40℃,並於該溫度持續攪拌2.5小時。於冰上加入6N HCl中止反應後,利用乙酸乙酯進行萃取,之後再以冰飽和K2CO3溶液及鹽水洗滌,以MgSO4乾燥處理,並真空濃縮。利用乙酸乙酯及正己烷(1:1→3:1,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的白色固體殘留物,以得到2.143克之白色固體化合物s9,產率73%,以及0.793克之白色固體化合物s10,產率21%:s9:Rf=0.13(乙酸乙酯:CH2Cl2=1:1(體積比));mp=99-100℃;[α]24 D -28.7(c 0.32,CHCl3);FT-IR(neat)vmax 3343,3064,2951,1719,1665,1602,1534,1447,1375,1313,1273,1117,1070,1026,994,895,753,713,618cm-1;1H NMR(400MHz,CDCl3)δ 8.06-8.00(m,4H),7.62-7.59(m,3H),7.50-7.43(m,4H),7.33-7.29(m,6H),7.25-7.23(m,2H),7.16(d,J=8.5Hz,1H),5.85(ddd,J=13.8,8.7,4.7Hz,1H),4.72(dd,J=11.8,2.4Hz,1H),4.57(d,J=5.6Hz,1H),4.52-4.44(m,4H),4.31-4.23(m,1H),4.15-4.11(m,1H),3.91(d,J=15.6Hz,1H),3.82(d,J=15.6Hz,1H),3.64-3.62(m,1H),3.57(s,31-1),2.89(dd,J=13.8,4.7Hz,1H),2.44(bs,1H),2.37 (dd,J=13.8,11.8Hz,1H);13C NMR(100MHz,CDCl3)δ172.6(C),171.0(C),168.5(C),167.3(C),166.6(C),136.4(C),135.7(CH),133.7(CH),133.6(CH),133.1(CH),130.2(CH),130.1(CH),129.9(CH),129.7(CH),129.7(CH),129.5(CH),129.1(CH),129.0(CH),128.6(CH),128.5(CH),128.4(CH),128.3(CH),127.8(CH),89.1(C),73.5(CH2),73.0(CH),69.4(CH),69.3(CH),68.9(CH),68.6(CH2),67.5(CH2),52.7(CH3),51.2(CH),38.0(CH2);HRMS-ESI[M+Na]+ Calcd for C39H39NO11SNa 752.2136,Found 752.2118;s10:Rf=0.45(乙酸乙酯:CH2Cl2=1:1(體積比));mp=115-117℃;[α]24 D -48.6(c 0.15,CHCl3);FT-IR(neat)vmax 3341,3065,1718,1603,1538,1449,1271,1172,1112,1071,1026,890,749,711cm-1;1H NMR(400MHz,CDCl3)δ 8.11-8.09(m,2H),8.03-8.01(m,2H),7.92-7.90(m,2H),7.64-7.59(m,2H),7.57-7.42(m,8H),7.41-7.36(m,2H),7.31-7.27(m,4H),7.22-7.19(m,2H),7.08-7.04(m,1H),5.91-5.84(m,1H),5.49(ddd,J=8.0,3.4,2.0Hz,1H),4.95(dd,J=12.3,2.0Hz,1H),4.66(dd,J=12.3,8.0Hz,1H),4.53-4.46(m,3H),4.29(d,J=10.3,8.4Hz,1H),4.07(dd,J=3.4,1.5Hz,1H),3.96-3.83(m,2H),3.56(s,3H),2.88(dd,J=13.8,4.8Hz,1H),2.40(dd,J=13.8,11.8Hz,1H);13C NMR(75MHz,CDCl3)δ173.1(C),171.0(C),168.7(C),166.6(C),166.4(C),166.0(C),136.5(C),136.2(CH),133.8(CH),133.6(CH),133.3(CH),132.9(CH),130.3(CH),130.2(CH),130.1(CH),130.0(CH),129.9(CH),129.8(CH), 129.6(CH),129.1(CH),128.9(CH),128,7(CH),128.6(CH),128.5(CH),128.3(CH),128.2(CH),127.8(CH),89.4(C),74.5(CH),74.3(CH),73.5(CH2),69.9(CH),68.9(CH),68.7(CH2),64.1(CH2),52.6(CH3),51.1(CH),37.8(CH2);HRMS-ESI[M+Na]+ Calcd for C46H43NO12SNa 856.2398,Found 856.2419.
(苯基4,9-二-O-苯甲醯基-5-苄氧乙醯胺基-3,5-二脫氧-7,8-二-O-甲基砒碇-2-硫基-D-丙三基-D-半乳-非-2-酮糖苷)甲酯(Methyl(phenyl 4,9-di-O-benzoyl-5-benzyloxyacetamido-3,5-dideoxy-7,8-di-O-picoloyl-2-thio-D-glycero-D-galacto-non-2-ulopyranoside)onate)(1h)
於0℃,將2-吡啶甲酸(0.758公克,6.15毫莫耳)、DMAP(0.030公克,0.25毫莫耳)及DCC(1.524公克,7.38毫莫耳)加至包含化合物s9(1.795公克,2.46毫莫耳)之無水二氯乙烷(12毫升)中。將混合物加熱至32℃,並於該溫度持續攪拌1小時。以矽藻土短墊過濾混合物後,進行真空濃縮。利用乙酸乙酯及正己烷(3:1→6:1,體積比)作為沖提液,以快速管柱層析於矽膠純化觀察到的黃色固體,得到2.126克之白色固體1b,產率92%:Rf=0.20(乙酸乙酯:CH2Cl2=4:1(體積比)); mp=139-140℃;[α]24 D +52.7(c 0.15,CHCl3);FT-IR(neat)vmax 3359,3063,3008,2956,1726,1685,1602,1585,1527,1452,1439,1275,1118,1027,994,893,751,712,619cm-1;1H NMR(400MHz,CDCl3)δ 8.78-8.75(m,2H),8.15(dt,J=7.9,1.0Hz,1H),8.04(dt,J=7.9,1.0Hz,1H),7.95-7.92(m,2H),7.87-7.83(m,3H),7.77(td,J=7.9,1.8Hz,1H),7.56-7.50(m,4H),7.49-7.42(m,2H),7.39-7.28(m,7H),7.21-7.18(m,2H),7.14-7.10(m,1H),6.81(d,J=10.3Hz,1H),6.03(t,J=2.3Hz,1H),5.71(ddd,J=11.4,10.0,4.8Hz,1H),5.66(dt,J=8.4,2.3Hz,1H),5.04(dd,J=10.9,2.5Hz,1H),4.95(dd,J=12.3,2.5Hz,1H),4.54-4.49(m,3H),4.45(q,J=10.0Hz,1H),3.90-3.64(m,5H),2.97(dd,J=13.9,4.8Hz,1H),2.32(dd,J=13.9,11.4Hz,1H);13C NMR(75MHz,CDCl3)δ170.5(C),168.5(C),166.1(C),165.7(C),164.3(C),164.0(C),150.1(CH),150.0(CH),147.6(C),147.3(C),137.2(CH),136.9(CH),136.9(CH),136.1(CH),133.3(CH),132.8(CH),129.8(CH),129.8(CH),129.5(CH),129.3(CH),129.2(CH),128.5(CH),128.3(CH),128.2(CH),127.7(CH),127.7(CH),127.6(CH),127.2(CH),126.9(CH),125.7(CH),125.5(CH),88.8(C),74.4(CH),73.2(CH),73.2(CH2),71.3(CH),70.1(CH),69.3(CH2),63.5(CH2),52.6(CH3),48.9(C),37.8(CH2);HRMS-ESI[M+Na]+ Calcd for C51H45N3O13SNa 962.2565,Found 962.2550.
1.2 將實施例1之唾液酸供體與一葡苷基受體耦合
本實施例係依據下述條件,分別將實施例1之唾液酸供體(即,化合物1a-e)分別與一葡苷基受體2耦合,單離產物(即,化合物3a-e及4a-e)後,以1H-NMR進行分析。表1闡述分析結果。
一般醣化流程:於室溫含氮環境中攪拌包含實施例1之唾液酸供體(1.0當量)、受體(1.2當量)及活化3Å粉末狀分子篩(每毫莫耳1.0公克)之無水CH2Cl2(0.05M)混合物1小時,以移除殘留水份。使反應混合物溫度降至-40℃後,加入N-碘琥珀醯亞胺(NIS)(2當量)及TfOH(1.5當量)。於該溫度持續攪拌直至薄層液相色層分析法(thin-layer liquid chromatography,TLC)顯示反應已完成。加入三乙胺中止反應後,以矽藻土短墊進行過濾。利用飽和Na2S2O3溶液及鹽水洗滌產物後,以MgSO4乾燥處理、過濾及真空濃縮產物。
雙醣3a及烯糖4a
甲基5-乙醯胺基-4,8,9-三-O-苯甲醯基-3,5-二脫氧-7-O-甲基砒碇-D-丙三基-D-半乳-非-2-ulopyranosylonate-(2→6)-甲基2,3,4-三-O-苄基-α-D-哌喃糖苷(Methyl 5-acetamido-4,8,9-tri-O-benzoyl-
3,5-dideoxy-7-O-picoloyl-D-glycero-D-galacto-non-2-ulopyranosylonate-(2→6)-methyl 2,3,4-tri-O-benzyl-α-D-哌喃糖苷)(3a);以及5-乙醯胺基-2,6-脫水-4,8,9-三-O-苯甲醯基-3,5-二脫氧-7-O-甲基砒碇-D-丙三基-D-半乳-非-2-酮酸甲酯(5-acetamido-2,6-anhydro-4,8,9-tri-O-benzoyl-3,5-dideoxy-7-O-picoloyl-D-glycero-D-galacto-non-2-enonate)(4a)
依據一般流程,將包含NIS(0.162公克,0.72毫莫耳)及TfOH(0.048公克,0.54毫莫耳)之CH2Cl2(7.0毫升)加入硫唾液酸苷1a(0.304公克,0.36毫莫耳)、哌喃糖苷受體2(0.201公克,0.43毫莫耳)及活化3Å粉末狀分子篩(0.360克)中,反應2小時。利用乙酸乙酯及CH2Cl2(1:2,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.243克之無色漿狀雙醣3a,產率57%,以及0.104克之無色漿狀烯糖4a,產率40%:3a:Rf=0.23(乙酸乙酯:CH2Cl2=1:2(體積比));FT-IR(neat)vmax 3064,3031,2934,1723,1685,1602,1452,1364,1312,1278,1214,1165,1114,1025,751,713cm-1;1H NMR(400MHz,CDCl3)δ 8.81-8.80(m,1H),8.70-8.69(m,1H),8.21(td,J=4.3,0.9Hz,1H),8.18-8.16(m,1H),8.00-7.85(m,8H),7.66(td,J=7.7,1.7Hz,1H),7.55-7.27(m,29H),6.01(t,J=1.5Hz,1H),5.94(td,J=5.6,1.6Hz,1H),5.89(dd,J=8.3,2.0Hz,1H), 5.58(dd,J=12.3,2.4Hz,1H),5.55-5.48(m,1H),5.42-5.38(m,1H),5.26-5.20(m,1H),5.02-4.63(m,11H),4.53(dd,J=10.9,2.0Hz,1H),4.40(q,J=10.3Hz,1H),4.32(dd,J=10.3,4.6Hz,1H),4.09(d,J=10.7Hz,1H),4.05-3.99(m,1H),3.95(d,J=10.7Hz,1H),3.77(s,3H),3.72(dd,J=9.5,3.5Hz,1H),3.67(dd,J=10.9,2.4Hz,1H),3.61(t,J=9.5Hz,1H),3.53(dd,J=9.9,3.5Hz,1H),3.38(s,3H),3.31(s,3H),2.85(dd,J=12.7,4.8Hz,1H),2.76(dd,J=12.9,5.0Hz,1H),2.08(dd,J=12.9,11.4Hz,1H),1.79(s,3H),1.77(s,3H);13C NMR(100MHz,CDCl3)δ170.2(C),167.4(C),166.4(C),166.2(C),166.0(C),165.9(C),164.0(C),150.1(CH),147.6(C),138.9(C),138.5(C),138.5(C),137.1(CH),133.3(CH),133.2(CH),133.0(CH),129.9(CH),129.7(CH),129.5(CH),129.4(CH),128.5(CH),128.4(CH),128.4(CH),128.4(CH),128.3(CH),128.3(CH),128.8(CH),128.0(CH),127.9(CH),127.9(CH),127.7(CH),127.7(CH),127.5(CH),127.0(CH),125.6(CH),98.3(C),97.9(CH),82.2(CH),80.3(CH),75.8(CH2),75.2(CH2),74.2(CH),73.2(CH2),72.1(CH),71.4(CH),69.8(CH),69.2(CH),63.5(CH2),62.4(CH2),55.0(CH),52.8(CH),49.7(CH),38.0(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C67H66N2O18Na 1209.4203,Found 1209.4214;4a:Rf=0.20(乙酸乙酯:CH2Cl2=1:2(體積比));[α]24 D -99.0(c 0.45,CHCl3);FT-IR(neat)vmax 3320,3063,3006,2931,2851,1727,1664,1593,1541,1445,1372,1269,1107,1022,756,713cm-1;1H NMR(400MHz,CDCl3)δ 8.74-8.71(m,1H),8.13(d,J=7.8Hz,1H),8.01-7.96(m,6H),7.82(t,J=7.8Hz,1H),7.59-7.30(m,10H),6.33(d,J=5.7Hz,1H),6.23-6.20(m,2H),6.09-6.06(m,1H),5.54(dd,J=5.7,3.9Hz,1H),5.30(dd,J=12.4,2.8Hz,1H),4.86(ddd,J=12.4,8.6,3.9Hz,1H),4.76-4.70(m,2H),3.80(s,3H),1.87(s,3H);13C NMR(100MHz,CDCl3)δ 170.1(C),166.2(C),166.1(C),165.4(C),163.8(C),161.8(C),150.0(CH),147.1(C),146.6(C),137.1(CH),133.5(CH),133.4(CH),133.0(CH),129.8(CH),129.8(CH),129.7(CH),129.6(CH),129.3(CH),128.5(CH),128.5(CH),128.3(CH),127.2(CH),125.6(CH),106.2(CH),74.3(CH),72.5(CH),70.3(CH),65.1(CH),63.2(CH2),52.6(CH3),45.2(CH),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C39H34N2O12Na 745.2004,Found 745.2005.
雙醣3b及烯糖4b
甲基5-乙醯胺基-4,7,9-三-O-苯甲醯基-3,5-二脫氧-8-O-甲基砒碇-D-丙三基-D-半乳-非-2-ulopyranosylonate-(2→6)-甲基2,3,4-三-O-苄基-α-D-哌喃糖苷(Methyl 5-acetamido-4,7,9-tri-O-benzoyl-3,5-dideoxy-8-O-picoloyl-D-glycero-D-galacto-
non-2-ulopyranosylonate-(2→6)-methyl 2,3,4-tri-O-benzyl-α-D-哌喃糖苷)(3b);以及5-乙醯胺基-2,6-脫水-4,7,9-三-O-苯甲醯基-3,5-二脫氧-8-O-甲基砒碇-D-丙三基-D-半乳-非-2-酮酸甲酯(methyl 5-acetamido-2,6-anhydro-4,7,9-tri-O-benzoyl-3,5-dideoxy-8-O-picoloyl-D-glycero-D-galacto-non-2-enonate)(4b)
依據一般流程,將包含NIS(0.027公克,0.12毫莫耳)及TfOH(0.008毫升,90.03毫莫耳)之CH2Cl2(1.2毫升)加入硫唾液酸苷1b(0.050公克,60.02微莫耳)、哌喃糖苷受體2(0.033公克,72.03微莫耳)及活化3Å粉末狀分子篩(0.060克),反應2小時。利用丙酮及Et2O(1:6,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.044克之無色漿狀雙醣3b,產率62%,以及0.010克之無色漿狀烯糖4b,產率24%:3b:Rf=0.43(丙酮:正己烷=1:1(體積比));FT-IR(neat)vmax 3306,2920,2854,1728,1641,1548,1458,1275,1084,751,711cm-1;1H NMR(400MHz,CDCl3)δ 8.76(d,J=4.2Hz,1H),8.07(d,J=7.8Hz,1H),7.97-7.90(m,6H),7.80(td,J=7.7,1.5Hz,1H),7.55-7.47(m,4H),7.39-7.24(m,18H),7.18-7.12(m,3H),5.99-5.89(m,2H),5.52(d,J=9.7Hz,1H),5.11-4.93(m,3H),4.86-4.75(m,3H),4.69-4.64(m,2H),4.49-4.45(m,2H),4.38(dd,J=10.2,3.7Hz,1H),4.27(q,J=10.2Hz,1H),4.11(dd,J=12.5,6.2Hz,1H),3.99(t,J=9.2Hz,1H), 3.84-3.31(m,1H),3.67(t,J=9.4Hz,1H),3.59(dd,J=10.4,1.7Hz,1H),3.54(dd,J=9.7,3.4Hz,1H),3.48(s,3H),3.38(s,3H),2.86(dd,J=12.7,4.5Hz,1H),2.10(t,J=12.7Hz,1H),1.79(s,3H);13C NMR(100MHz,CDCl3)δ170.1(C),168.0(C),166.5(C),166.1(C),165.6(C),163.9(C),149.7(CH),148.2(C),139.1(C),138.5(C),138.4(C),137.1(CH),133.5(CH),133.4(CH),133.1(CH),130.0(CH),129.9(CH),129.9(C),129.6(C),129.4(C),128.6(CH),128.6(CH),128.5(CH),128.5(CH),128.4(CH),128.3(CH),128.1(CH),128.0(CH),127.7(CH),127.7(CH),127.0(CH),125.7(CH),125.7(CH),99.2(C),98.3(CH),82.2(CH),79.6(CH),75.9(CH2),74.9(CH2),73.5(CH2),72.8(CH),70.1(CH),69.7(CH),69.7(CH),68.2(CH),63.7(CH2),63.1(CH2),55.3(CH3),52.6(CH3),49.8(CH),38.5(CH2),30.5(CH3),23.3(CH3);HRMS-ESI[M+Na]+ Calcd for C67H66N2O18Na 1209.4203,Found 1209.4181;4b:Rf=0.38(丙酮:正己烷=1:1(體積比));[α]30 D +126.7(c 0.20,CHCl3);FT-IR(neat)vmax 3277,3066,3016,2960,2925,2855,1729,1671,1593,1546,1445,1368,1262,1104,1029,989,908,857,755,711,610cm-1;1H NMR(400MHz,CDCl3)δ8.67(d,J=4.4Hz,1H),8.09(d,J=7.4Hz,2H),8.06(d,J=7.9Hz,1H),7.98(t,J=6.4Hz,4H),7.81(t,J=7.7Hz,1H),7.59(t,J=7.4Hz,1H),7.54-7.45(m,5H),7.40-7.35(m,4H),7.00(bs,1H),6.17(dd,J=5.0,2.3Hz,1H),6.09(d,J=2.3Hz,1H),6.07-6.04(m,1H),5.93(dd,J=8.8,2.3Hz,1H),5.25(dd, J=12.4,2.9Hz,1H),4.94(dd,J=10.7,1.4Hz,1H),4.72-4.60(m,2H),3.77(s,3H),1.84(s,3H);13C NMR(100MHz,CDCl3)δ170.4(C),166.6(C),166.3(C),165.7(C),164.1(C),161.9(C),149.9(CH),147.2(C),145.5(C),137.6(CH),133.5(CH),133.5(CH),133.2(CH),130.1(CH),130.0(CH),129.8(CH),129.6(C),129.5(C),128.7(CH),128.6(CH),128.5(CH),127.6(CH),125.9(CH),109.0(CH),76.9(CH),73.4(CH),70.2(CH),68.9(CH),63.0(CH2),52.6(CH3),47.2(CH),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C39H34N2O12Na 745.2004,Found 745.2005.
雙醣3c及烯糖4c
甲基5-乙醯胺基-7.8-二-O-苯甲醯基-3,5-二脫氧-4,9-二-O-甲基砒碇-D-丙三基-D-半乳-非-2-ulopyranosylonate-(2→6)-甲基2,3,4-三-O-苄基-α-D-哌喃糖苷(Methyl 5-acetamido-7.8-di-O-benzoyl-3,5-dideoxy-4,9-di-O-picoloyl-D-glycero-D-galacto-non-2-ulopyranosylonate-(2→6)-methyl 2,3,4-tri-O-benzyl-α-D-glucopyranoside)(3c);以及5-乙醯胺基-2,6-脫水-7,8-二-O-苯甲醯基-3,5-二脫
氧-4,9-二-O-甲基砒碇-D-丙三基-D-半乳-非-2-酮酸甲酯(methyl 5-acetamido-2,6-anhydro-7,8-di-O-benzoyl-3,5-dideoxy-4,9-di-O-picoloyl-D-glycero-D-galacto-non-2-enonate)(4c)
依據一般流程,將包含NIS(0.043公克,0.19毫莫耳)及TfOH(0.013毫升,0.14毫莫耳)之CH2Cl2(1.9毫升)加入硫唾液酸苷1c(0.080公克,0.10毫莫耳)、葡哌喃糖苷受體2(0.060公克,0.12毫莫耳)及活化3Å粉末狀分子篩(0.120克),反應2小時。利用乙酸乙酯及丙酮/CH2Cl2(1/1,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.049克之無色漿狀雙醣3c,產率43%,以及0.034克之無色漿狀烯糖4c,產率43%:3c:Rf=0.13(乙酸乙酯);FT-IR(neat)vmax 3326,2922.2854,1733,1678,1546,1446,1366,1280,1095,752,708,613cm-1;1H NMR(400MHz,CDCl3)δ 8.79(d,J=4.3Hz,1H),8.76(d,J=4.9Hz,1H),8.74(d,J=4.7Hz,2H),8.12(dd,J=7.4Hz,2H),8.06-7.93(m,12H),7.81-7.77(m,4H),7.70(td,J=7.8,1.5Hz,1H),7.62-7.30(m,37H),7.23-7.22(m,6H),6.29(dd,J=8.6,7.6Hz,1H),5.97-5.86(m,4H),5.75-5.68(m,1H),5.55(dd,J=11.9,3.7Hz,1H),5.44-5.37(m,1H),4.97-4.58(m,20H),4.38(dd,J=12.2,5.8Hz,1H),4.34(q,J=10.0Hz,1H),4.22(q,J=9.8Hz,1H),4.15(dd,J=10.4,4.5Hz,1H),4.05(d,J=10.5Hz,1H),4.00-3.92(m,2H),3.85(d,J=11.0Hz,1H),3.80-3.78(m,2H),3.73-3.70(m, 4H),3.61-3.57(m,2H),3.52-3.47(m,3H),3.43(s,3H),3.34(s,6H),2.82(dd,J=12.6,4.9Hz,1H),2.72(dd,J=13.1,5.0Hz,1H),2.16-2.08(m,2H),1.81(s,3H),1.78(s,3H);13C NMR(100MHz,CDCl3)δ170.5(C),170.4(C),170.0(C),167.4(C),166.2(C),165.7(C),165.6(C),165.5(C),164.6(C),164.5(C),164.5(C),164.3(C),150.2(CH),150.1(CH),150.0(CH),147.6(C),147.5(C),147.4(C),147.2(C),139.0(C),138.8(C),138.6(C),138.6(C),138.4(C),138.3(C),137.4(CH),137.3(CH),137.2(CH),137.1(CH),133.3(CH),133.1(CH),130.0(CH),129.9(CH),129.8(C),129.7(C),129.5(C),128.6(CH),128.5(CH),128.4(CH),128.4(CH),128.3(CH),128.2(CH),128.1(CH),128.0(CH),127.9(CH),127.8(CH),127.7(CH),127.7(CH),127.6(CH),127.3(CH),127.2(CH),125.7(CH),125.6(CH),125.4(CH),125.0(CH),99.1(C),98.2(C),98.1(CH),98.0(CH),82.3(CH),82.0(CH),80.1(CH),79.7(CH),77.6(C),77.4(C),75.8(CH2),75.8(CH2),75.2(CH2),74.9(CH2),73.9(CH),73.4(CH2),73.2(CH2),72.5(CH),71.5(CH),71.0(CH),70.8(CH),70.7(CH),69.6(CH),69.5(CH),69.4(CH),69.4(CH),64.0(CH2),63.7(CH2),63.6(CH2),62.6(CH2),55.2(CH3),55.1(CH3),52.8(CH3),52.5(CH3),50.1(CH),49.9(CH),38.3(CH2),37.9(CH2),23.2(CH3),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C66H65N3O18Na 1210.4155,Found 1210.4135;4c:Rf=0.13(乙酸乙酯);[α]30 D +162.8(c 0.31,CHCl3);FT-IR(neat)vmax 3318,3064,3006,2923, 2853,1732,1673,1578,1545,1444,1369,1254,1100,1035,991,907,855,755,711,614cm-1;1H NMR(400MHz,CDCl3)δ8.85(d,J=4.0Hz,1H),8.72(d,J=4.3Hz,1H),8.10(d,J=8.0Hz,4H),7.94(d,J=8.0Hz,2H),7.83(td,J=8.0,1.2Hz,1H),7.78(td,J=8.0,1.2Hz,1H),7.60(t,J=7.5Hz,1H),7.53-7.43(m,5H),7.35(t,J=7.7Hz,2H),7.13(d,J=9,3Hz,1H),6.17(dd,J=8.4,2.5Hz,1H),6.11(t,J=3.4Hz,1H),6.07(d,J=2.6Hz,1H),6.00-5.97(m,1H),5.19(dd,J=12.2,3.4Hz,1H),5.06(dd,J=10.5,2.8Hz,1H),4.75(dd,J=12.2,5.9Hz,1H),4.53(q,J=9.5Hz,1H),3.65(s,3H),1.89(s,3H);13C NMR(100MHz,CDCl3)δ171.8(C),165.6(C),165.6(C),164.9(C),164.7(C),161.8(C),150.1(CH),150.0(CH),147.4(C),147.0(C),145.7(C),137.5(CH),137.3(CH),133.4(CH),133.3(CH),130.0(CH),129.8(CH),129.5(C),129.3(C),128.6(CH),128.4(CH),127.5(CH),127.3(CH),125.9(CH),125.4(CH),108.3(CH),76.6(CH),71.4(CH),71.2(CH),69.1(CH),64.2(CH2),52.5(CH3),47.0(CH),23.1(CH3);HRMS-ESI[M+Na]+ Calcd for C38H33N3O12Na 746.1956,Found 746.1952.
雙醣3d及烯糖4d
甲基5-乙醯胺基-4,9-二-O-苯甲醯基
-3,5-二脫氧-7,8-二-O-甲基砒碇-D-丙三基-α-D-半乳-非-2-ulopyranosylonate-(2→6)-甲基2,3,4-三-O-苄基-α-D-哌喃糖苷(Methyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-picoloyl-D-glycero-α-D-galacto-non-2-ulopyranosylonate-(2→6)-methyl 2,3,4-tri-O-benzyl-α-D-glucopyranoside)(3d);以及5-乙醯胺基-2,6-脫水-4,9-二-O-苯甲醯基-3,5-二脫氧-7,8-二-O-甲基砒碇-D-丙三基-D-半乳-非-2-酮酸甲酯(methyl 5-acetamido-2,6-anhydro-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-picoloyl-D-glycero-D-galacto-non-2-enonate)(4d)
依據一般流程,將包含NIS(0.016公克,71.94微莫耳)及TfOH(0.002毫升,53.96微莫耳)之CH2Cl2(0.7毫升)加入硫唾液酸苷1d(0.030公克,35.97微莫耳)、葡哌喃糖苷受體2(0.020公克,43.16微莫耳)及活化3Å粉末狀分子篩(0.040克),反應2小時。利用乙酸乙酯作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.030克之白色固體雙醣3d,產率70%,以及0.007克之白色固體烯糖4d,產率27%:3d:Rf=0.28(乙酸乙酯);mp=111-112℃;[α]24 D +52.4(c 0.35,CHCl3);FT-IR(neat)vmax 3330,3062,3008,2926,1730,1678,1579,1446,1366,1280,1121,752,709cm-1;1H NMR(400MHz,CDCl3)δ 8.73 (d,J=4.4Hz,1H),8.66(d,J=4.2Hz,1H),8.06(d,J=7.8Hz,1H),7.98-7.90(m,5H),7.79(td,J=7.7,1.6Hz,1H),7.64(td,J=7.7,1.7Hz,1H),7.54-7.46(m,3H),7.42-7.29(m,16H),7.26-7.11(m,4H),6.01-5.95(m,2H),5.66(d,J=9.8Hz,1H),5.12-5.05(m,1H),5.03-4.64(m,7H),4.60-4.57(m,1H),4.49(d,J=10.8Hz,1H),4.37(dd,J=10.6,3.9Hz,1H),4.29(q,J=10.2Hz,1H),4.07(dd,J=12.5Hz,1H),4.00(t,J=9.2Hz,1H),3.83-3.80(m,1H),3.66(t,J=9.4Hz,1H),3.61(dd,J=10.6,1.7Hz,1H),3.54(dd,J=9.7,3.4Hz,1H),3.45(s,3H),3.38(s,3H),2.85(dd,J=12.7,4.6Hz,1H),2.11(t,J=12.7Hz,1H),1.79(s,3H);13C NMR(100MHz,CDCl3)δ170.4(C),168.0(C),166.4(C),166.0(C),164.0(C),163.8(C),150.1(CH),149.5(CH),148.0(C),147.4(C),139.1(C),138.7(C),138.3(C),137.2(CH),136.9(CH),133.4(CH),133.0(CH),129.9(CH),129.9(CH),129.8(C),129.4(C),128.6(CH),128.5(CH),128.4(CH),128.4(CH),128.2(CH),128.0(CH),128.0(C),127.7(CH),127.6(CH),127.6(CH),127.1(CH),127.0(CH),125.7(CH),125.5(CH),99.1(C),98.2(CH),82.1(CH),79.6(CH),75.8(CH2),74.8(CH2),73.4(CH2),72.6(CH),70.2(CH),69.7(CH),69.7(CH),68.8(CH),63.7(CH2),62.6(CH2),55.2(CH3),52.5(CH3),49.7(CH),38.5(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C66H65N3O18Na 1210.4155,Found 1210.4101.
雙醣3e
甲基5-乙醯胺基-4,7,8,9-四-O-苯甲醯基-3,5-二脫氧-D-丙三基-D-半乳-非-2-ulopyranosylonate-(2→6)-甲基2,3,4-三-O-苄基-α-D-哌喃糖苷(Methyl 5-acetamido-4,7,8,9-tetra-O-benzoyl-3,5-dideoxy-D-glycero-D-galacto-non-2-ulopyranosylonate-(2→6)-methyl 2,3,4-tri-O-benzyl-α-D-glucopyranoside)(3e)
依據一般流程,將包含NIS(0.162公克,0.72毫莫耳)及TfOH(0.048公克,0.54毫莫耳)之CH2Cl2(7.0毫升)加入硫唾液酸苷1e(0.300公克,0.36毫莫耳)、葡哌喃糖苷受體2(0.201公克,0.43毫莫耳)及活化3Å粉末狀分子篩(0.467克),反應2小時。利用乙酸乙酯及正己烷(1:2→1:1,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.398克之白色固體雙醣3e,產率90%:Rf=0.25(乙酸乙酯:正己烷=1:2(體積比));FT-IR(neat)vmax 3395,3065,3031,2935,1724,1602,1527,1452,1364,1315,1270,1215,1171,1100,1071,1025,754,713cm-1;1H NMR(400MHz,CDCl3)δ 8.14-8.12(m,2H),7.99-7.97(m,2H),7.95-7.92(m,2H),7.88-7.85(m,2H), 7.62-7.29(m,27H),5.93(t,J=1.7Hz,1H),5.90-5.87(m,1H),5.60-5.57(m,2H),5.52-5.45(m,1H),5.02-4.79(m,7H),4.70-4.63(m,2H),4.35(q,J=10.2Hz,1H),4.09-4.00(m,2H),3.96-3.94(m,1H),3.78(s,3H),3.72(dd,J=9.6,3.6Hz,1H),3.29(s,3H),2.74(dd,J=12.9,5.0Hz,1H),2.05(dd,J=12.9,11.9Hz,1H),1.77(s,3H);13C NMR(75MHz,CDCl3)δ170.1(C),167.4(C),166.5(C),166.4(C),166.1(C),165.5(C),138.9(C),138.6(C),138.5(C),133,3(CH),133.3(CH),133.2(CH),133.0(CH),130.0(CH),129.9(CH),129.8(CH),129.8(CH),129.5(CH),128.6(CH),128.5(CH),128.4(CH),128.4(CH),128.3(CH),128.1(CH),128.0(CH),127.8(CH),127.7(CH),127.6(CH),98.3(CH),98.0(CH),82.2(CH),80.1(CH),75.8(CH2),75.2(CH2),74.8(CH),73,2(CH2),72.2(CH),70.4(CH),69.9(CH),69.4(CH),63.5(CH2),62.5(CH2),55.0(CH3),52.8(CH3),49.6(CH),38.0(CH2),23.2(CH3);HRMS-ESI[M+Na]+ Calcd for C68H67NO18Na 1208.4250,Found 1208.4226.
表1之數據證明唾液酸供體1d與受體2之最佳醣化反應溫度為-40℃,其中產物3d的產率約為70%,且具有優異的立體選擇性(僅α),伴隨產率為27%之產物4d。唾液酸供體1b亦具有高度α-立體選擇性(α/β=8:1)及中等產率之雙醣3b(62%),伴隨產率為24%之產物4b。相較之下,唾液酸化合物1a及1c則對雙醣3a(α/β=1:1.2,產率為57%)及3c(α/β=1.3:1,產率為43%)不具顯著的立體選擇性,其中化合物4a及4c的產率分別約為40%及49%。唾液酸供體1e可高產率(90%)及低α-立體選擇性(α/β=1:3.3) 地製備雙醣3e。
1.3 篩選最佳醣化條件
為分析實施例1.2之耦合反應的最佳醣化條件,依據與實施例1,2所述之相似流程(除了調整溶劑及溫度)來耦合唾液酸供體1d及受體2;之後單離及分析產物3d及4d。表2總結分析結果。
如表2結果所示,當將耦合溫度由-40℃提升至-20℃時,會降低α-立體選擇性(僅α與α/β=5.3:1)及產率(70%與24%)。相較之下,若進一步將耦合溫度降低至-60℃時,除了反應時間較長(約5小時),雙醣3d的產率(69%)及α-立體選擇性(僅α)會維持與溫度為-40℃時相似的結果。
於CH3CN或CHCl3中進行耦合反應,而非CH2Cl2,則依然可維持優異的α-立體選擇性(僅α),惟雙醣3d的產率卻不盡理想(56%或20%)。當將溶劑 置換為甲苯時,則完全不會發生醣化反應。
實施例2 篩選實施例1之唾液酸供體的醣苷基受體
本實施例將利用實施例1.3確認之最佳條件來製備不同的糖基受體,並測試其與唾液酸供體1d耦合的效率。表3總結代表性結果。
表3之數據指出,唾液酸供體1d及一級醇5a的醣耦合可最高產率地(97%)製備雙醣6a,其為單一α-立體異構物。一級醇5b亦具有優異的(98%)產率;然而,其α-立體選擇性卻不盡理想(α/β=2.8:1)。另外二種一級醇5c及5d僅能中等產率地製備雙醣6c及6d(分別為68%及73%),其具有優異的α-立體選擇性(僅α)。當以一級糖5e作為受體時,未發現對應的雙醣6e,而僅有烯糖副產物4d(99%)。
實施例3 以實施例1確認之唾液酸供體及最佳醣化條件來合成Hp-s1
為確認本揭示內容的應用性,依據流程圖I所述流程,以實施例1確認之方法及唾液酸供體來合成天然神經節苷脂Hp-s1。一般來說,以實施例1確認之最佳條件來進行S-噻唑基(S-thiazolyl,STaz)受體8與唾液酸供體1d的唾液酸化反應,其中以NIS/TfOH作為促進劑,且於-40℃之包含粉末狀3Å分子篩(MS-3Å)的CH2Cl2中進行耦合反應。耦合反應會生成α-立體異構物雙醣9(產率為76%),之後於0℃之包含促進劑(例如,AgOTf)之無水CH2Cl2中,與具保護基之乙醯植物鞘胺醇11進行耦合反應,僅能製得具保護基之Hp-s1 12的β-變旋異構物,產率為25%。另一方面,若先於-10℃,以包含Cu(OAc)2之MeOH處理化合物9移除甲基砒碇基後,將製得的雙醣10與具保護基之乙醯植物鞘胺醇11進行醣化反應,在與上述用以製備雙醣 9之相同條件下,產率可提升至93%。Hp-s1相似物13的產率亦高達87%,且具有優異的立體選擇性(僅β)。接著使具保護基之Hp-s1相似物12及13進行去保護基反應,以製備神經節苷脂Hp-s1,其產率分別為75%及89%。
2-噻唑啉基(甲基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-7,8-二-O-甲基砒碇-D-丙三基-α-D-半乳-2-吡喃壬酮糖甲酯)-(2→6)-2-O-乙醯基-3,4-二-O-苄基-β-D-哌喃糖苷(2-Thiazolinyl(methyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-picoloyl-D-glycero-α-D-galacto-2-nonulopyranoylonate
)-(2→6)-2-O-acetyl-3,4-di-O-benzyl-β-D-glucopyranoside)(9)
依一般流程,將包含NIS(0.271公克,1.20毫莫耳)及TfOH(0.080毫升,0.90毫莫耳)之CH2Cl2(12毫升)加入硫唾液酸苷1d(0.500公克,0.60毫莫耳)、葡哌喃糖苷受體8(0.360公克,0.72毫莫耳)及活化3Å粉末狀分子篩(0.600克),反應1小時c利用乙酸乙酯及丙酮/CH2Cl2(1/1,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.560克之無色漿狀雙醣9,產率76%:Rf=0.43(丙酮:CH2Cl2=1:1(體積比));[α]30 D +72.1(c 0.13,CHCl3);FT-IR(neat)vmax 3318,2922,2854,1736,1674,1576,1445,1370,1280,1237,1122,1078,753,710,612cm-1;1H NMR(400MHz,CDCl3)δ 8.74(d,J=4.4Hz,1H),8.67(d,J=4.4Hz,1H),8.07(d,J=7.8Hz,1H),7.99-7.92(m,5H),7.80(td,J=7.7,1.4Hz,1H),7.62(td,J=7.7,1.6Hz,1H),7.53-7.46(m,3H),7.41-7.20(m,16H),6.05-6.04(m,2H),5.72(d,J=9.6Hz,1H),5.27(d,J=10.4Hz,1H),5.14(ddd,J=12.7,8.8,4.6Hz,1H),5.08(dd,J=10.4,9.2Hz,1H),4.97-4.29(m,9H),4.27-4.09(m,2H),3.83-3.69(m,3H),3.60(dd,J=9.6,2.2Hz,1H),3.49(s,3H),3.32(t,J=8.1Hz,2H),2.87(dd,J=12.7,4.6Hz,1H),2.09(t,J=12.7Hz,1H),1.95(s,3H),1.79(s,3H);13C NMR(100MHz,CDCl3)δ170.4(C),169.5(C),167.6(C),166.1(C),165.8(C),163.8(C),163.7(C),163.5(C),149.9(CH),149.5(CH),147.7(C),147.1(C),138.0(C),138.0(C), 137.0(CH),136.9(CH),133.2(CH),132.8(CH),129.6(CH),129.6(C),129.3(C),128.3(CH),128.3(CH),128.2(CH),127.9(CH),127.7(CH),127.6(CH),126.9(CH),126.9(CH),125.5(CH),125.4(CH),99.1(C),84.0(CH),83.1(CH),78.4(CH),77.0(CH),75.1(CH2),74.8(CH2),72.7(CH),71.2(CH),70.5(CH),70.1(CH),69.1(CH),64.0(CH2),63.4(CH2),62.8(CH2),52.5(CH3),49.3(CH),41.8(CH2),38.1(CH2),34.9(CH2),26.9(CH2),24.8(CH2),23.0(CH3),20.8(CH3);HRMS-ESI[M+Na]+ Calcd for C63H62N4O18S2Na 1249.3394,Found 1249.3369.
(甲基5-乙醯胺基-4,9-二-O-苯甲醯基-3,5-二脫氧-7,8-二-O-甲基砒碇-D-丙三基-α-D-半乳-2-吡喃壬酮糖甲酯)-(2→6)-2-O-乙醯基-3,4-二-O-苄基-β-D-葡萄呱喃糖苷-(1→1)-(2S,3S,4R)-3,4-O-鄰異亞丙基-2-十八醯氨基十八烷-1,3,4-三醇((Methyl 5-acetamido-4,9-di-O-benzoyl-3,5-dideoxy-7,8-di-O-picoloyl-D-glycero-α-D-galacto-2-nonulopyranoylonate)-(2→6)-2-O-acetyl-3,4-di-O-benzyl-β-D-glucopyranosyl-(1→1)-(2S,3S,4R)-3,4-O-isopropylidenyl-
2-octadecanoylaminooctadecane-1,3,4-triol)(12)
在含氮環境中,攪拌溶於無水CH2Cl2(0.9毫升)中的唾液酸供體9(0.110公克,0.09毫莫耳)、受體11(0.067公克,0.11毫莫耳)及活化3Å粉末狀分子篩(0.124克),以移除殘留水份。使反應混合物降溫至0℃後,加入AgOTf(0.500公克,0.27毫莫耳)。於該溫度持續攪拌直到TLC顯示完全反應;將三乙胺加入反應混合物後,以矽藻土短墊進行過濾。以冰飽和Na2S2O3溶液及鹽水洗滌過濾物,利用MgSO4乾燥處理後,進行過濾及真空濃縮。利用丙酮及CH2Cl2(1/2,體積比)作為沖提液,以快速管柱層析於矽膠純化製得的淡黃色漿狀殘留物,得到0.038克之無色漿狀雙醣12,產率25%:Rf=0.30(乙酸乙酯);[α]30 D +45.5(c 0.21,CHCl3);FT-IR(neat)vmax 3306,3065,2925,2855,1736,1670,1544,1458,1371,1279,1120,753,709,614cm-1;1H NMR(400MHz,CDCl3)δ8.75(d,J=4.0Hz,1H),8.67(d,J=4.0Hz,1H),8.07(d,J=7.8Hz,1H),7.98-7.91(m,5H),7.80(td,J=7.8,1.7Hz,1H),7.63(td,J=7.8,1.7Hz,1H),7.53-7.47(m,3H),7.42-7.19(m,15H),6.10-6.03(m,2H),5.85(d,J=8.2Hz,1H),5.70(d,J=9.6Hz,1H),5.15-5.09(m,1H),4.97-4.62(m,6H),4.50(dd,J=10.9,1.4Hz,1H),4.36-4.28(m,4H),4.12-4.03(m,3H),3.96(dd,J=10.0,3.6Hz,1H),3.77-3.72(m,2H),3.64(t,J=9.1Hz,1H),3.57(dd,J=10.0,1.8Hz,1H),3.49(s,3H),2.84(dd,J=12.7,4.4Hz,1H),2.14-2.11(m,3H),1.95(s,3H),1.80(s,3H),1.58-1.47(m,4H),1.35(s,3H), 1.25(s,55H),0.88(t,J=6.8Hz,6H);13C NMR(100MHz,CDCl3)δ172.7(C),170.3(C),169.8(C),167.9(C),166.5(C),166.1(C),164.1(C),164.0(C),150.2(CH),149.8(CH),148.1(C),148.1(C),147.4(C),138.4(C),138.2(C),137.1(CH),137.0(CH),133.5(CH),133.1(CH),130.0(CH),129.9(CH),129.8(C),129.4(C),128.6(CH),128.6(CH),128.5(CH),128.4(CH),128.2(CH),128.0(CH),127.9(CH),127.8(CH),127.1(CH),127.0(CH),125.8(CH),125.6(CH),107.9(C),101.3(CH),99.3(C),82.9(CH),77.9(CH),77.6(CH),76.3(CH),75.2(CH2),75.0(CH2),74.3(CH),73.5(CH),72.7(CH),69.9(CH),69.8(CH),69.2(CH2),69.1(CH),63.7(CH2),63.0(CH2),52.7(CH3),50.0(CH),48.2(CH),38.5(CH2),36.9(CH2),32.1(CH2),29.9(CH2),29.8(CH2),29.8(CH2),29.7(CH2),29.7(CH2),29.6(CH2),29.6(CH2),29.5(CH2),29.2(CH2),28.1(CH3),26.7(CH2),25.8(CH3),25.8(CH2),23.3(CH3),22.8(CH2),21.2(CH3),14.3(CH3);HRMS-ESI[M+Na]+ Calcd for C99H134N4O22Na 1753.9382,Found 1753.9383.
合成標的化合物Hp-s1於0℃,將異亞丙基縮醛12(0.050公克,28.90微莫耳)溶於80%之AcOH溶 液(10毫升)中,並於85℃持續攪拌混合物4小時。使反應混合物與甲苯共蒸發。不需進一步純化,於室溫將製得的無色漿狀二醇溶於MeOH(10毫升)後,加入20%Pd(OH)2/C(0.003克)。於室溫及含氫(50psi)環境中,持續攪拌反應混合物1小時。以SiO2/矽藻土短墊移除Pd(OH)2/C後,利用MeOH洗滌過濾物。之後於0℃真空濃縮過濾物。不需進一步純化,將製得的無色漿狀四醇溶於無水MeOH(0.27毫升)後,加入MeONa(1毫克,0.027毫莫耳)。於室溫持續攪拌3小時後,加入H2O(0.15毫升)。在完成皂化反應後,以Dowex 50w X 8[H+]中和溶液。過濾樹脂,並以MeOH/CH2Cl2(3:1,體積比)洗滌。於減壓環境中濃縮過濾物,以得到白色固體殘留物。使22毫克之Hp-s1再結晶(MeOH/CH2Cl2/EtOAc)後,得到一白色固體化合物,三步驟產率75%:Rf=0.23(MeOH:CH2Cl2=1:1(體積比));[α]24 D -1.3(c 0.20,MeOH:CHCl3=3:1);FT-IR(neat)vmax 3373,2921,2853,1639,1552,1481,1372,1314,1204,1116,1080,1040,760cm-1;1H NMR(400MHz,CD3OD:CDCl3=3:1)δ 4.26(d,J=7.7Hz,1H),4.17-4.05(m,3H),3.86-3.82(m,2H),3.77-3.61(m,6H),3.59-3.51(m,3H),3.45-3.32(m,3H),3.21(t,J=8.4Hz,1H),2.74(dd,J=12.3,4.2Hz,1H),2.21(t,J=7.4Hz,2H),2.01(s,3H),1.76(t,J=12.3Hz,1H),1.61-1.39(m,4H),1.28(s,52H),0.89(t,J=6.8Hz,6H);13C NMR(100MHz,CD3OD:CDCl3=3:1)δ 174.6(C),174.2(C),170.5(C),103.4(CH),98.2(C),76.1(CH),74.9(CH),73.6(CH),73.5(CH),73.4(CH),71.8(CH),71.1(CH),69.3(CH),69.1(CH2),68.7(CH),67.3(CH),63.2(CH2),62.3(CH2), 52.5(CH),50.4(CH),40.2(CH2),36.2(CH2),31.8(CH2),31.8(CH2),31.1(CH2),29.7(CH2),29.6(CH2),29.6(CH2),29.6(CH2),29.6(CH2),29.6(CH2),29.5(CH2),29.5(CH2),29.4(CH2),29.4(CH2),29.2(CH2),29.2(CH2),25.9(CH2),25.8(CH2),22.5(CH2),21.8(CH3),13.6(CH3);HRMS-ESI[M-H]- Calcd for C53H99N2O17 1035.6949,Found 1035.6939.
雖然上文實施方式中揭露了本發明的具體實施例,然其並非用以限定本發明,本發明所屬技術領域中具有通常知識者,在不悖離本發明之原理與精神的情形下,當可對其進行各種更動與修飾,因此本發明之保護範圍當以附隨申請專利範圍所界定者為準。
Claims (9)
- 如請求項1所述之唾液酸供體,其中在式(I)之唾液酸供體中,R 1及R 2分別是苯甲醯基,且R 3是乙醯基。
- 如請求項4所述之方法,其中步驟(a)是於介於-20℃到-60℃的溫度下,在一選自CH 3CN、CH 3Cl及CH 2Cl 2所組成之群組的溶劑中進行耦合。
- 如請求項5所述之方法,其中步驟(a)是於-40℃的溫度,在CH 2Cl 2中進行耦合。
- 如請求項4所述之方法,其中步驟(a)是以一粉末分子篩來進行該耦合反應。
- 如請求項4所述之方法,其中在式(I)之唾液酸供體中,R 1及R 2分別是苯甲醯基,且R 3是乙醯基。
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