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    • 4. 发明申请
    • SYNTHESIS OF OLIGONUCLEOTIDES
    • 合成寡核苷酸
    • WO2008141682A1
    • 2008-11-27
    • PCT/EP2007/062660
    • 2007-11-21
    • GIRINDUS AGLANGE, MeinolfHOHLFELD, AndreasSCHÖNBERGER, AndreasKIRCHHOFF, ChristinaGRÖSSEL, Olaf
    • LANGE, MeinolfHOHLFELD, AndreasSCHÖNBERGER, AndreasKIRCHHOFF, ChristinaGRÖSSEL, Olaf
    • C07H21/00
    • C07H21/00
    • A method for preparing an oligonucleotide comprising the steps of a) providing a hydroxyl containing compound having the formula (1), wherein B is a heterocyclic base and the radicals R 2 , R 3 and R 5 are as defined in the description; b) reacting said compound with a phosphitylating agent in the presence of an activator having the formula (I) (activator I)), wherein R = alkyl, cycloalkyl, aryl, aralkyl, heteroalkyl, heteroaryl; R 1 , R 2 = either H or form a 5 to 6-membered ring together; X 1 , X 2 = independently either N or CH; Y = H or Si(R 4 ) 3 , with R 4 = alkyl, cycloalkyl, aryl, aralkyl, heteroalkyl, heteroaryl; B = deprotonated acid; to prepare a phosphitylated compound; c) reacting said phosphitylated compound without isolation with a second compound having the formula (1), wherein R 5 , R 3 , R 2 , B are independently selected, but have the same definition as above in the presence of an activator II selected from the group of imidazole imidazolium salts and mixtures thereof.
    • 一种制备寡核苷酸的方法,包括以下步骤:a)提供具有式(1)的含羟基化合物,其中B是杂环碱基,R 2,R 3, SUB>和R 5 5如说明书中所定义; b)在具有式(I)的活化剂(活化剂I))的存在下使所述化合物与磷酸酯化剂反应,其中R =烷基,环烷基,芳基,芳烷基,杂烷基,杂芳基; R 1,R 2,H或H或一起形成5至6元环; X 1,X 2独立地是N或CH; Y = H或Si(R 4)3,其中R 4 =烷基,环烷基,芳基,芳烷基,杂烷基,杂芳基; B =去质子酸 制备磷酸化化合物; c)使所述磷化物化合物与所述式(1)的第二化合物分离而不分离,其中R 5,R 3,R 2, B独立选择,但在选自咪唑咪唑盐及其混合物的活化剂II的存在下具有与上述相同的定义。
    • 5. 发明申请
    • SYNTHESIS OF OLIGONUCLEOTIDES
    • 合成寡核苷酸
    • WO2006094963A1
    • 2006-09-14
    • PCT/EP2006/060489
    • 2006-03-06
    • GIRINDUS AGLANGE, MeinolfSCHÖNBERGER, AndreasHOHLFELD, AndreasGRÖSSEL, OlafKIRCHHOFF, ChristineLINK, Fritz
    • LANGE, MeinolfSCHÖNBERGER, AndreasHOHLFELD, AndreasGRÖSSEL, OlafKIRCHHOFF, ChristineLINK, Fritz
    • C07H21/00
    • C07H21/00Y02P20/55
    • A method for preparing an oligonucleotide comprising the steps of a) providing a hydroxyl containing compound having the formula (A), wherein B is a heterocyclic base and i) R 2 is H, a protected 2'-hydroxyl group, F, a protected amino group, an O-alkyl group, an O-substituted alkyl, a substituted alkylamino or a C4 '-O2 ' methylen linkage; R 3 is OR' 3 , NHR" 3 , NR" 3 R' 3 , wherein R 3 is a hydroxyl protecting group, a protected nucleotide or a protected oligonucleotide, R" 3 , R" 3 are independently amine protecting groups, and R 5 is OH or ii) R 2 is H, a protected 2'-hydroxyl group, F, a protected amino group, an O-alkyl group, an O-substituted alkyl, a substituted alkylamino or a C4'-O2 'methylen linkage; R 3 is OH and R 5 is OR' 5 and R' 5 is a hydroxyl protecting group, a protected nucleotide or a protected oligonucleotide or iii) R 2 is OH, R 3 is OR' 3 , NHR" 3 , NR" 3 R'" 3 , wherein R 3 is a hydroxyl protecting group, a protected nucleotide or a protected oligonucleotide, R" 3 , R" 3 are independently amine protecting groups, and R 5 is OR' 5 and R' 5 is a hydroxyl protecting group, a protected nucleotide or a protected oligonucleotide b) reacting said compound with a phosphitylating agent in the presence of an activator having the formula (I) (activator I) wherein R = alkyl, cycloalkyl, aryl, aralkyl, heteroalkyl, heteroaryl; R 1 , R 2 = either H or form a 5 to 6-membered ring together; X 1 , X 2 = independently either N or CH; Y = H or Si(R 4 ) 3 , with R 4 = alkyl, cycloalkyl, aryl, aralkyl, heteroalkyl, heteroaryl; B = deprotonated acid to prepare a phosphitylated compound; c) reacting said phosphitylated compound without isolation with a second compound having the formula (A) wherein R 5 , R 3 , R 2 , B are independently selected, but have the same definition as above in the presence of an activator II different from activator I.
    • 一种制备寡核苷酸的方法,包括以下步骤:a)提供具有式(A)的含羟基化合物,其中B是杂环碱基,i)R 2是H,被保护的2'- 羟基,F,被保护的氨基,O-烷基,O-取代的烷基,取代的烷基氨基或C 4'-O 2'亚甲基键; R 3是OR 3,NHR“3”,“NR”3 R“3” 其中R 3是羟基保护基,被保护的核苷酸或被保护的寡核苷酸,R“3,R”3个独立地是 胺保护基,R 5是OH或ii)R 2是H,被保护的2'-羟基,F,被保护的氨基,O-烷基 基团,O-取代的烷基,取代的烷基氨基或C4'-O2'亚甲基键; R 3是OH,R 5是OR 5,并且R 5是羟基保护基, 受保护的核苷酸或受保护的寡核苷酸或iii)R 2是OH,R 3是OR 3,NHR“3, 其中R 3是羟基保护基,受保护的核苷酸或受保护的寡核苷酸,R 3,R 3,R 3,R 3, “3”,R“3”3“独立地为胺保护基,R 5为OR'5和R' b)羟基保护基,受保护的核苷酸或受保护的寡核苷酸b)使所述化合物与磷酸化剂在存在下具有式(I)的活化剂(活化剂I)(其中R =烷基 ,环烷基,芳基,芳烷基,杂烷基,杂芳基; R 1,R 2 = H或一起形成5至6元环; X 1, X 2独立地是N或CH; Y = H或Si(R 4)3,其中R 1, 4 = alk 芳基,芳烷基,杂烷基,杂芳基; B =脱质子酸以制备磷酸化化合物; c)使所述磷化物化合物与不与其分离的具有式(A)的第二化合物反应,其中R 5,R 3,R 2,B, 在不同于活化剂I的活化剂II的存在下,它们具有与上述相同的定义。
    • 7. 发明申请
    • PURIFICATION OF OLIGONUCLEOTIDES
    • 纯化寡核苷酸
    • WO2006079660A1
    • 2006-08-03
    • PCT/EP2006/050503
    • 2006-01-30
    • GIRINDUS AGLANGE, MeinolfGRÖSSEL, OlafLINK, FritzSCHÖNBERGER, AndreasHOHLFELD, Andreas
    • LANGE, MeinolfGRÖSSEL, OlafLINK, FritzSCHÖNBERGER, AndreasHOHLFELD, Andreas
    • C07H21/00
    • C07H21/00
    • A method for purifying a protected oligonucleotide comprising the steps of: a) - providing a solution of the protected oligonucleotide in at least one solvent A having a boiling point below the boiling point of a solvent B, - heating the solution at a temperature of at least 30°C and below the boiling point of the at least solvent A, - adding solvent B until precipitation of a material is visible in the solution, said solvent B being an alcohol having 1 to 6 C-atoms or a diol having 2 to 6 C-atoms, - allowing the solution to cool down under stirring until formation of a supernatant and a residue, - removing the supernatant or b) - providing solvent B, said solvent B being an alcohol having 1 to 6 C- atoms or a diol having 2 to 6 C-atoms, - heating solvent B at a temperature above 30°C and below the boiling point of solvent B, - adding a solution of a protected oligonucleotide in at least one solvent A until precipitation of a material is visible in the solution, - allowing the solution to cool down under stirring until formation of a supernatant and a residue, - removing the supernatant.
    • 一种用于纯化受保护的寡核苷酸的方法,包括以下步骤:a) - 将受保护的寡核苷酸的溶液提供在沸点低于溶剂B的沸点的至少一种溶剂A中, - 在 至少30℃且低于至少溶剂A的沸点, - 加入溶剂B,直到在溶液中可见材料沉淀,所述溶剂B是具有1至6个C原子的醇或具有2至 6个C原子, - 允许溶液在搅拌下冷却直到形成上清液和残余物, - 去除上清液或b)提供溶剂B,所述溶剂B是具有1-6个C原子的醇或 具有2至6个C原子的二醇, - 在高于30℃且低于溶剂B的沸点的温度下加热溶剂B, - 在至少一种溶剂A中加入受保护的寡核苷酸的溶液,直到材料沉淀可见 在解决方案中, - 允许soluti 在搅拌下冷却直到形成上清液和残余物, - 除去上清液。