DEVELOPMENT OF CANCER THERAPEUTICS BASED ON REGULATION OF CHROMATIN STRUCTURE AND FUNCTION

基于染色质结构和功能调控的癌症治疗方法的开发

• 批准号: 12219205
• 负责人: YOSHIDA Minoru
• 金额: $ 83.01万
• 依托单位: RIKEN (2002-2004)The University of Tokyo (2000-2001)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12219205/
• 关键词: ACETYLATION; HDAC; MICROTUBULE; CYCLIC TETRAPEPTIDES; DRUG DESIGN; THIOL; FK228; SCOP; プロドラッグ; エンドサイトーシス; トラポキシン; ヒストンデアセチラーゼ; 酸化還元; トリコスタチン; チューブリン; ヒストン; グルタチオン; 細胞周期; ラディシコール; ヒストンアセチル化; 転写制御; クロマチン

We have identified histone deacetylases (HDAC) as the molecular target of trichostatin A (TSA) and trapoxin (TPX), microbial metabolites that inhibit mammalian cell cycle. Recently, we have shown that FK228, a potent anti-cancer agent under the phase II clinical trials in the US, also specifically inhibits HDAC. Since accumulating evidence suggests that HDAC is a novel molecular target for anti-cancer drugs, we developed new types of HDAC inhibitors in this project The X-ray crystallographic studies suggested that TSA inhibits HDAC by chelating the active-site zinc with its hydroxamic acid group, while TPX covalently modifies an active site residue via its epoxiketone moiety. We synthesized a hybrid inhibitor by coupling the cyclic tetrapeptide of TPX with the hydroxamic acid of TSA, named CHAP CHAP was shown to be a potent HDAC inhibitor. In contrast to TSA or TPX, CHAP was highly stable in serum and a derivative (CHAP31) showed potent antitumor activity in the xenograft models. FK228 has an intramolecular disulfide. We showed that FK228 is a natural prodrug, which can be converted to the reduced form with a thiol group interacting with the active-site zinc by cellular reducing activity. We therefore designed a new type of inhibitors by introducing the thiol group into the TPX-like cyclic tetrapeptide inhibitors named SCOP. SCOP was normally oxidized and inactivated but became active when it is reduced in cells. The HDAC family includes more than ten species of enzymes. TSA inhibited almost all the enzymes but TPX failed to inhibit HDAC6. By using the differential sensitivity, we identified that HDAC6 is the tubulin deacetylase. Since each member of HDAC family may have each distinct function, it is important to develop subtype-specific inhibitors. During the course of screening forth subtype-specific inhibitors, we found a SCOP derivative that specifically inhibits HDAC4.

我们已确定组蛋白脱乙酰酶 (HDAC) 是曲古抑菌素 A (TSA) 和曲古霉素 (TPX) 的分子靶标，这两种微生物代谢物可抑制哺乳动物细胞周期。最近，我们发现在美国进行II期临床试验的强效抗癌药物FK228也能特异性抑制HDAC。由于越来越多的证据表明HDAC是抗癌药物的新型分子靶标，因此我们在该项目中开发了新型HDAC抑制剂。X射线晶体学研究表明，TSA通过与其异羟肟酸基团螯合活性位点锌来抑制HDAC ，而 TPX 通过其环氧酮部分共价修饰活性位点残基。我们通过将 TPX 的环状四肽与 TSA 的异羟肟酸偶联合成了一种混合抑制剂，命名为 CHAP CHAP，它被证明是一种有效的 HDAC 抑制剂。与 TSA 或 TPX 相比，CHAP 在血清中高度稳定，并且其衍生物 (CHAP31) 在异种移植模型中显示出有效的抗肿瘤活性。 FK228具有分子内二硫键。我们证明FK228是一种天然前药，可以通过细胞还原活性将其转化为带有硫醇基团与活性位点锌相互作用的还原形式。因此，我们通过在类TPX环四肽抑制剂中引入硫醇基团，设计了一种新型抑制剂，命名为SCOP。 SCOP通常被氧化并失活，但当它在细胞中被还原时就会变得活跃。 HDAC 家族包括十多种酶。 TSA 抑制几乎所有酶，但 TPX 未能抑制 HDAC6。通过使用差异敏感性，我们确定 HDAC6 是微管蛋白脱乙酰酶。由于 HDAC 家族的每个成员可能具有各自不同的功能，因此开发亚型特异性抑制剂非常重要。在筛选亚型特异性抑制剂的过程中，我们发现了一种特异性抑制HDAC4的SCOP衍生物。

项目成果

FK228 (depsipeptide) as a natural prodrug that inhibits class I histone deacetylases.

• 发表时间: 2002-09
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: R. Furumai;Akihisa Matsuyama;N. Kobashi;Kun-Hyung Lee;M. Nishiyama;H. Nakajima;A. Tanaka;Y. Komatsu;N. Nishino;Minoru Yoshida;S. Horinouchi

Bradney, C., et al.: "Regulation of E2A activities by histone acetyltransferases in B lymphocyte development."J.Biol.Chem.. 278. 2370-2376 (2003)

Bradney, C., 等人：“B 淋巴细胞发育中组蛋白乙酰转移酶对 E2A 活性的调节。”J.Biol.Chem.. 278. 2370-2376 (2003)

Komatsu et al.: "Cyclic Hydroxamic-acid-containing Peptide 31, a potent synthetic histone deacetylase inhibitor with antitumor activity"Cancer Res.. 61. 4459-4466 (2001)

Komatsu 等人：“含有环异羟肟酸的肽 31，一种具有抗肿瘤活性的有效合成组蛋白脱乙酰酶抑制剂”Cancer Res.. 61. 4459-4466 (2001)

Akakura,S. et al.: "A role Hsc70 in regulating nucleo-cytoplasmic transport of a temperature-sensitive p53 (p53^<Vall35>)."J.Biol.Chem.. 276(印刷中). (2001)

Akakura, S. 等人：“Hsc70 在调节温度敏感 p53 (p53^<Vall35>) 中的作用。J.Biol.Chem.. 276（出版中）。”

Verdel,A. et al.: "Active maintenance of mHDA2/mHDAC6 histone-deacetylase in the cytoplasm."Curr.Biol.. 10. 747-749 (2000)

韦尔德尔，A.