α-tert-butyl γ-methyl N'-<4--N-methylamino>benzoyl>-L-glutamate | 79648-88-7

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 羧酸及其衍生物
• 英文名称: α-tert-butyl γ-methyl N'-<4--N-methylamino>benzoyl>-L-glutamate
• 英文别名: methotrexate α-tert-butyl γ-methyl ester；1-O-tert-butyl 5-O-methyl (2S)-2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]pentanedioate
• CAS: 79648-88-7
• 化学式: C₂₅H₃₂N₈O₅
• 分子量: 524.58
• InChiKey: GYZQDMZIFVWTDV-KRWDZBQOSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  1.2
• 重原子数：
  38
• 可旋转键数：
  12
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.4
• 拓扑面积：
  189
• 氢给体数：
  3
• 氢受体数：
  12

反应信息

• 作为反应物：
  描述：

  α-tert-butyl γ-methyl N'-<4--N-methylamino>benzoyl>-L-glutamate 在 三氟化硼乙醚 作用下， 以 1,2-二氯乙烷 为溶剂， 反应 18.0h， 以88%的产率得到甲氨蝶呤gamma-甲酯
  参考文献：
  名称：

  Synthesis of methotrexate-antibody conjugates by regiospecific coupling and assessment of drug and antitumor activities

  摘要：

  In order to increase the retention of drug activity, regiospecific coupling has been used to synthesize conjugates of methotrexate (MTX, 1) with normal rabbit IgG (NRG) and a mouse anti-human renal cancer monoclonal IgG (Dal K-20). MTX gamma-methyl ester (4) was produced either by selective esterification of MTX or by coupling of 4-amino-4-deoxy-N10-methylpteroic acid (2) with suitable glutamic acid derivatives. The MTX gamma-methyl ester (4) was then converted to the corresponding hydrazide 6. An amide-linked conjugate was formed when the MTX gamma-hydrazide (6) was converted to reactive acylating species 7 by using tert-butyl nitrite or trifluoroacetaldehyde, which were reacted with nucleophilic centers, presumably epsilon-amino groups, in native IgG. A hydrazone-linked conjugate was formed when MTX gamma-hydrazide (6) was reacted directly with IgG that had first been oxidized with periodate to form polyaldehyde IgG. The regiospecifically synthesized conjugates were somewhat more effective inhibitors in vitro of dihydrofolate reductase and of colony formation by human renal cancer (Caki-1) cells than were control nonregiospecific conjugates.

  DOI：

  10.1021/jm00131a003
• 作为产物：
  描述：

  L-谷氨酸-5-甲酯 在 高氯酸 、 氰基磷酸二乙酯 、 三乙胺 作用下， 反应 74.03h， 生成 α-tert-butyl γ-methyl N'-<4--N-methylamino>benzoyl>-L-glutamate
  参考文献：
  名称：

  Methotrexate analogs. 14. Synthesis of new .gamma.-substituted derivatives as dihydrofolate reductase inhibitors and potential anticancer agents

  摘要：

  The gamma-tert-butyl ester (1), gamma-hydrazide (2), gamma-n-butylamide (3), and gamma-benzylamide (4) derivatives of methotrexate (MTX) were synthesized from 4-amino-4-deoxy-N10-methylpteroic acid (APA) and the appropriate blocked L-glutamic acid precursors with the aid of the peptide bond forming reagent diethyl phosphorocyanidate. The affinity of these side chain modified products for dihydrofolate reductase (DHFR) from Lactobacillus casei and L1210 mouse leukemic cells was determined spectrophotometrically or by competitive radioligand binding assay, and their cytotoxicity was evaluated against L1210 leukemic cells in culture. The results provide continuing support for the view that the "gamma-terminal region" of the MTX side chain is an attractive site for molecular modification of this anticancer agent.

  DOI：

  10.1021/jm00144a016

文献信息

• Synthesis of methotrexate-containing heterodimeric molecules
  申请人：GPC Biotech, Inc.

  公开号：US07230101B1

  公开（公告）日：2007-06-12

  The present invention relates to novel compositions of methotrexate-containing heterodimeric probe molecules, also known as chemical inducers of dimerization (CID), useful in three-hybrid assays. The invention further relates to synthesis of said compositions and their intermediates. Another aspect of the invention is a method for using the heterodimeric probe molecules described herein in drug screens to identify potential protein targets to a given ligand, optimize protein-ligand interactions, or identify potential ligands for a given protein target. In certain embodiments, the invention contemplates the synthesis of the following methotrexate-containing heterodimeric probe:

  本发明涉及含有甲氧苄氨甲酸的异二聚体探针分子的新型组合物，也称为二聚体诱导剂（CID），在三杂交分析中有用。本发明还涉及合成所述组合物及其中间体的方法。本发明的另一个方面是使用本文所述的异二聚体探针分子在药物筛选中的方法，以识别给定配体的潜在蛋白靶标，优化蛋白质-配体相互作用或识别给定蛋白靶标的潜在配体。在某些实施方式中，本发明考虑合成以下含有甲氧苄氨甲酸的异二聚体探针：
• Synthesis, Cleavage Profile, and Antitumor Efficacy of an Albumin-Binding Prodrug of Methotrexate that is Cleaved by Plasmin and Cathepsin B
  作者：André Warnecke、Iduna Fichtner、Gretel Saß、Felix Kratz

  DOI：10.1002/ardp.200700025

  日期：2007.8

  The albumin‐bound form of the prodrug was shown to be efficiently cleaved by cathepsin B and plasmin as well as in an ovarian carcinoma homogenate (OVCAR‐3) liberating a methotrexate‐lysine derivative. In an OVCAR‐3 xenograft model, the prodrug at a dose of 4×15 mg/kg methotrexate equivalents demonstrated distinctly superior antitumor efficacy compared to free methotrexate at a dose of 4×100 mg/kg [T/C(%)

  组织蛋白酶 B 和纤溶酶是细胞内或细胞外蛋白酶，在多种实体瘤中过度表达。为了利用这两种蛋白酶作为前药肿瘤特异性裂解的分子靶点，开发了一种结合白蛋白的甲氨蝶呤制剂，其中结合了肽序列 D-Ala-Phe-Lys 作为蛋白酶底物。由于在实体瘤中被动积累，白蛋白是细胞抑制剂的合适载体。通过将肽接头 EMC-D-Ala-Phe-Lys (Boc)-Lys-OH（EMC = ε-马来酰亚胺己酸）与 α-叔丁基保护的甲氨蝶呤的 γ-COOH 基团偶联来进行合成。在切割保护基团并在反相 HPLC 上纯化后，获得了高度水溶性的甲氨蝶呤 - 肽衍生物，可快速且选择性地与人血清白蛋白结合。前药的白蛋白结合形式被证明可以被组织蛋白酶 B 和纤溶酶以及在卵巢癌匀浆 (OVCAR-3) 中有效裂解，释放出甲氨蝶呤 - 赖氨酸衍生物。在 OVCAR ‐3 异种移植模型中，与剂量为 4 × 100 mg / kg [T /
• The use of Tris-Lipidation to modify drug cytotoxicity in multidrug resistant cells expressing P-glycoprotein or MRP1
  作者：Ross A Davey、Mary W Davey、Karen V Cullen、Xanthe E Wells、Craig L Francis、Hua-Ming Williams、Qi Yang、Minoo J Moghaddam、Fred Widmer、Robert G Whittaker

  DOI：10.1038/sj.bjp.0704983

  日期：2002.12

  Increasing the lipophilicity is a strategy often used to improve a compound's cellular uptake and retention but this may also convert it into a substrate for an ATP‐dependent transporter such as P‐glycoprotein or the multidrug resistance‐associated protein (MRP1), which are involved in cellular efflux of drugs. Tris‐Lipidation of compounds is a convenient way of modifying drug lipophilicity and generating an array of derivatives with diverse properties. To determine the effect of Tris‐Lipidation on a drug's cytoxicity in multidrug resistant cells, various glycyl‐Tris‐mono‐ (GTP1), di‐ (GTP2) and tri‐palmitate (GTP3) derivatives were prepared of the cancer chemotherapeutic drugs chlorambucil and methotrexate, and of the anti‐HIV drug AZT. The cytotoxicity of these derivatives and their parent compounds was determined in the CEM/VLB100 cells with increased P‐glycoprotein expression, the CEM/E1000 cells that overexpress MRP1 and the parent, drug‐sensitive CCRF‐CEM cells. Increasing the lipophilicity of AZT increased its cytotoxicity in the sensitive CCRF‐CEM parental cell line while decreased cytotoxicity was observed for the methotrexate derivatives. For the chlorambucil derivatives, both increased (GTP1) and decreased (GTP2) cytotoxicity occurred in the CCRF‐CEM cells. With the exception of AZT‐GTP1, all GTP1 and GTP2 derivatives of chlorambucil, methotrexate and AZT had decreased cytotoxicity in the P‐glycoprotein‐expressing CEM/VLB100 cells while chlorambucil‐GTP1, methotrexate‐GTP2 and methotrexate‐GTP3 were the only compounds with decreased cytotoxicity in the MRP1‐overexpressing CEM/E1000 cells. The number of palmitate residues, the position of derivatisation and the type of linkage all may affect the P‐glycoprotein and MRP1 substrate properties. Tris‐Lipidation may therefore provide a useful way of manipulating the pharmacokinetic properties of drugs. British Journal of Pharmacology (2002) 137, 1280–1286. doi:10.1038/sj.bjp.0704983

  增加疏水性是改善化合物细胞摄取和保留的常见策略，但也可能引发化合物成为ATP依赖性转运蛋白（如P-糖蛋白或多药耐药相关蛋白（MRP1））的底物，这些蛋白参与药物的细胞外排。将化合物进行三酰化是调节药物疏水性的一种简便方法，能够生成一系列具有不同特性的衍生物。 为了研究药物三酰化对多重耐药细胞毒性的影响，制备了抗癌化疗药物卡奇霉素和甲氨蝶呤以及抗HIV药物AZT的甘氨酰-Tris-单（GTP1）、双（GTP2）和三棕榈酰（GTP3）衍生物。在具有P-糖蛋白高表达的CEM/VLB100细胞、MRP1高表达的CEM/E1000细胞及其亲本药物敏感的CCRFR-CEM细胞中，研究了这些衍生物及其母体化合物的细胞毒性。 AZT的疏水性增加提高了其在敏感的CCRFR-CEM亲本细胞系中的细胞毒性，而甲氨蝶呤衍生物的细胞毒性有所降低。对于卡奇霉素衍生物，在CCRFR-CEM细胞中，GTP1增加细胞毒性，GTP2降低。除了AZT-GTP1，所有GTP1和GTP2的卡奇霉素、甲氨蝶呤和AZT衍生物在P-糖蛋白表达的CEM/VLB100细胞中细胞毒性降低，而在MRP1过表达的CEM/E1000细胞中仅有卡奇霉素-GTP1、甲氨蝶呤-GTP2 和甲氨蝶呤-GTP3 的细胞毒性降低。 棕榈酸残基的数量、修饰的位置和连接方式均可能影响P-糖蛋白和MRP1底物特性。 因此，药物的三酰化可能提供一种有效手段，用于调节药物的药代动力学特性。 英国药理学杂志 (2002) 137, 1280–1286。doi:10.1038/sj.bjp.0704983
• Tight binding ligand approach to oligosaccharide-grafted protein
  作者：Kiichiro Totani、Ichiro Matsuo、Yukishige Ito

  DOI：10.1016/j.bmcl.2004.01.106

  日期：2004.5

  A novel type of artificial glycoprotein was developed, by using dihydrofolate reductase (DHFR) and methotrexate (MTX) as a protein-ligand pair. Various oligosaccharides linked to MTX were shown to bind tightly with DHFR and afforded oligosaccharide-grafted protein, which could be isolated easily by lectin beads. (C) 2004 Elsevier Ltd. All rights reserved.
• KRALOVEC, J.;SPENCER, G.;BLAIR, A. H.;MAMMEN, M.;SINGH, M.;GHOSE, T., J. MED. CHEM., 32,(1989) N1, C. 2426-2431
  作者：KRALOVEC, J.、SPENCER, G.、BLAIR, A. H.、MAMMEN, M.、SINGH, M.、GHOSE, T.

  DOI：——

  日期：——