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    • 3. 发明授权
    • Vitamin B12 conjugates with EPO, analogues thereof and pharmaceutical
compositions
    • 维生素B12与EPO的缀合物,其类似物和药物组合物
    • US5548064A
    • 1996-08-20
    • US64873
    • 1993-05-24
    • Gregory J. Russell-JonesSteven W. Westwood
    • Gregory J. Russell-JonesSteven W. Westwood
    • A61K31/714A61K38/00A61K47/48C07K1/113C07K14/505C07K14/535C07K17/02C07K17/08C07K17/10C07H23/00C07K2/00
    • A61K47/48169A61K47/48023A61K47/48176C07K14/505C07K14/535C07K17/02A61K38/00
    • The invention describes complexes between VB.sub.12 analogues and either GCSF or EPO that retain both significant affinity for intrinsic factor (IF) in the VB.sub.12 portion of the complex and significant bioactivity of the GCSF or EPO portion of the complex. The invention also concerns a process for the synthesis of these complexes. This is achieved at least in part, by using a spacer compound, which is linked covalantly between the VB.sub.12 portion and the GCSF or EPO. The complexes preferably have the formulaV--X--A--Y--ZwhereinV is vitamin B.sub.12 or a vitamin B.sub.12 analogue, or derivative, bonded to X either through a carboxylate group pendant to the corrin nucleus of VB.sub.12 or through the central cobalt atom or to a functional group introduced onto the VB.sub.12 molecule, X is selected from: --NHNH--, --NH--, --O--, --S--, --SS--or --CH.sub.2 --, and A is an optionally substituted, saturated or unsaturated, branched or linear, C.sub.1-50 alkylene, cycloalkylene or aromatic group, optionally with one or more carbons within the linear chain being replaced with N, O or S, and wherein the optional substituents are selected from carbonyl, carboxy, hydroxy, amino and other groups, and Y is the covalent linkage between A and Z where Y is selected from --NHCO--, --CONH--, --CONHNHCO--, --N.dbd.N--, --N.dbd.CH--, --NHCH.sub.2 --, --NHN.dbd.CH--, --NHNHCH.sub.2 --, --SS--, --SCH.sub.2 --, --CH.sub.2 S--, --NHCRNH--, --COO--, --OCO--, and R is O, S or NH.sub.2, and Z is GCSF or EPO. The invention also describes reagents that can be used as probes for the detection of buried thiol groups of a protein or peptide, said reagent comprising a complex of either vitamin B.sub.12 (or an analogue thereof) or more generally of any instrumentally or visually detectable label, covalently linked to a diradical spacer, said spacer having a terminal reactive group capable of forming a disulphide bond with a free thiol in said protein or peptides.
    • 本发明描述了VB12类似物和GCSF或EPO之间的复合物,其在复合物的VB12部分中保留对内在因子(IF)的显着亲和力,并且复合物的GCSF或EPO部分具有显着的生物活性。 本发明还涉及合成这些络合物的方法。 这通过使用在VB12部分和GCSF或EPO之间共价连接的间隔化合物至少部分地实现。 络合物优选具有式V-XAY-Z,其中V是维生素B12或维生素B12类似物或衍生物,其通过悬浮于VB12的corrin核的羧酸酯基团或通过中心钴原子或引入的官能团 在VB12分子上,X选自:-NHN - , - NH - , - O - , - S - , - S - 或-CH 2 - ,A是任选取代的饱和或不饱和的支链或直链C1 -50个亚烷基,亚环烷基或芳基,任选与一个或多个直链内的碳被N,O或S取代,其中任选的取代基选自羰基,羧基,羟基,氨基和其它基团,Y是 其中Y选自-NHCO - , - CONH - , - CONHNHCO - , - N = N - , - N = CH - , - NHCH 2 - , - NHN = CH - , - NHNHCH 2 - , - -SS - , - CH 2 - , - CH 2 S - , - NHCRNH-,-COO-,-OCO-,R是O,S或NH 2,Z是GCSF或EPO。 本发明还描述了可用作检测蛋白质或肽的掩埋硫醇基团的探针的试剂,所述试剂包含维生素B12(或其类似物)或更通常的任何仪器上或视觉上可检测的标记物的复合物, 共价连接到双基间隔物上,所述间隔物具有能与所述蛋白质或肽中的游离硫醇形成二硫键的末端反应基团。