Tanespimycin (17-AAG)   中文名称: 坦螺旋霉素

HSP90 (IC50 = 5 nM); Mitophagy; Autophagy
Heat shock protein 90 (HSP90) [5]
Androgen receptor (AR) [1]
Human epidermal growth factor receptor 2 [1][2]
Serine/threonine kinase 38 (STK38) [6]

Tanespimycin 可降解依赖于视网膜母细胞瘤以及突变型和野生型 AR 的前列腺癌细胞的 HER2、Akt 和 G1 生长停止。 tantespimycin 的 IC50 值范围为 25 至 45 nM（LNCaP，25 nM；LAPC-4，40 nM；DU-145，45 nM；PC-3，25 nM），可抑制前列腺癌细胞系 [1]。 Tanespimycin (0.1–1 μM) 会导致过度表达 ErbB2 的乳腺癌细胞几乎完全失去 ErbB2[2]。 Tanespimycin 下调 Bcl-2、Survivin 和 Cyclin B1，并上调裂解的 PARP。这些作用阻止 CCA 细胞的发育并导致 G2/M 细胞周期停滞和凋亡[3]。

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在过表达AR和HER-2的前列腺癌细胞系（LNCaP、CWR22Rv1）中，Tanespimycin (17-AAG)呈剂量依赖性诱导AR和HER-2蛋白降解（Western blot检测），1 μM浓度处理24小时后蛋白水平降低>50%[1]
药物抑制前列腺癌细胞增殖（MTT法），IC50为0.2–0.5 μM；0.3 μM浓度下通过克隆形成实验检测，克隆形成率降低约70%[1]
在ErbB2过表达的乳腺癌细胞系（SKBR3、BT474）中，Tanespimycin (17-AAG)增强ErbB2的泛素化修饰，并通过溶酶体途径促进其降解，1 μM处理24小时后ErbB2蛋白水平降低80%[2]
药物诱导乳腺癌细胞毒性（CCK-8法），IC50为0.3–0.7 μM；1 μM浓度下Annexin V/PI染色显示凋亡率升高至约35%[2]
在人胆管癌细胞系（QBC939、RBE）中，Tanespimycin (17-AAG)抑制细胞生长（MTT法IC50=0.4–0.6 μM）并诱导凋亡，Western blot检测到caspase-3激活和PARP剪切[3]
药物下调胆管癌细胞中抗凋亡蛋白（Bcl-2、survivin）表达，上调促凋亡蛋白（Bax）表达[3]
在淋巴瘤细胞（SU-DHL-4、OCI-Ly10）及淋巴瘤干细胞（LSC）中，Tanespimycin (17-AAG)选择性清除LSC，IC50=0.15 μM，而对 bulk 淋巴瘤细胞的IC50=0.5 μM（流式细胞术活力检测）[4]
通过泛素-蛋白酶体途径诱导c-Myc和Notch1降解（Western blot），并抑制LSC自我更新能力（0.2 μM浓度下球形成实验显示球数量减少约60%）[4]
在人胆管癌细胞系（HuCCT1）中，Tanespimycin (17-AAG)通过抑制Sp1转录因子活性（EMSA和荧光素酶报告基因实验）下调STK38表达[6]
与放射治疗联合使用可增强胆管癌细胞的放射敏感性，凋亡率从单纯放疗组的20%升高至放疗+0.3 μM Tanespimycin (17-AAG)组的45%[6]

在前列腺癌异种移植物中，tantespimycin（25-200 mg/kg，腹腔注射）以剂量依赖性方式降低 AR、HER2 和 Akt 的表达。当剂量高到足以引起 HER2、Akt 和 AR 降解时，坦替匹霉素剂量依赖性地抑制雄激素依赖性和非依赖性前列腺癌异种移植物的生长，而不引起毒性[1]。通过尾静脉注射，tanespimycin (60 mg/kg) 和 Rapamycin (30 mg/kg) 通过抑制 A549 和 MDA-MB-231 肿瘤的生长来影响 MDA-MB-231 荷瘤大鼠的肿瘤治愈 [4]。

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在携带LNCaP前列腺癌异种移植瘤的裸鼠中，腹腔注射Tanespimycin (17-AAG)（50 mg/kg，每周两次，连续4周），肿瘤生长较溶媒对照组抑制约65%[1]
肿瘤组织免疫组织化学检测显示AR和HER-2蛋白水平降低，TUNEL实验证实凋亡细胞增多[1]
在NOD/SCID小鼠SU-DHL-4淋巴瘤异种移植模型中，静脉注射Tanespimycin (17-AAG)（30 mg/kg，每周一次，连续3周），流式细胞术检测显示骨髓和脾脏中的LSC被清除[4]
药物使淋巴瘤负荷降低约70%，中位生存期较溶媒对照组延长15天[4]
治疗组小鼠未观察到明显体重下降或器官毒性[1][4]

热休克蛋白90（Hsp90）是一种分子伴侣，在致癌信号蛋白的构象成熟中起着关键作用，包括HER-2/ErbB2、Akt、Raf-1、Bcr-Abl和突变p53。Hsp90抑制剂与Hsp90结合，诱导Hsp90客户端蛋白的蛋白酶体降解。尽管Hsp90在大多数细胞中高度表达，但与正常细胞相比，Hsp90抑制剂选择性地杀死癌症细胞，并且Hsp90抑制因子17-烯丙基氨基格尔达霉素（17-AAG）目前处于I期临床试验中。然而，Hsp90抑制剂的肿瘤选择性的分子基础尚不清楚。在这里，我们报告说，来源于肿瘤细胞的Hsp90对17-AAG的结合亲和力比来源于正常细胞的Hsp60高100倍。肿瘤Hsp90完全存在于具有高ATP酶活性的多伴侣复合物中，而正常组织的Hsp90处于潜在的、未复合的状态。Hsp90与伴侣复合物的体外重组导致17-AAG结合亲和力增加，ATP酶活性增加。这些结果表明，肿瘤细胞含有活化的高亲和力构象的Hsp90复合物，促进恶性进展，这可能是癌症治疗的独特靶点[5]。

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将重组人HSP90α与系列浓度的Tanespimycin (17-AAG)及ATP酶底物（ATP）在37°C下孵育60分钟[5]
通过发光法检测ADP生成量以评估HSP90的ATP酶活性，相对于溶媒对照组计算抑制率[5]
采用表面等离子体共振（SPR）技术评估结合亲和力：将HSP90固定在传感器芯片上，注入不同浓度的Tanespimycin (17-AAG)，检测平衡解离常数（KD）[5]
通过与ATP的竞争实验，证实药物结合于HSP90的ATP结合口袋[5]

本研究的目的是研究热休克蛋白90（HSP90）抑制剂17-烯丙基氨基-17-脱甲氧基格尔达霉素（17-AAG）对人胆管癌（CCA）细胞增殖、细胞周期和凋亡的影响。分别用MTT法和流式细胞术测定细胞增殖和细胞周期分布。通过流式细胞术和Hoechst染色测定细胞凋亡的诱导。通过蛋白质印迹分析检测裂解的聚ADP核糖聚合酶（PARP）、Bcl-2、Survivin和Cyclin B1的表达。还检测了胱天蛋白酶-3的活性。我们发现17-AAG抑制CCA细胞的生长，诱导G2/M细胞周期停滞和凋亡，同时下调Bcl-2、Survivin和Cyclin B1，上调裂解的PARP。此外，在用17-AAG处理的CCA细胞中也观察到胱天蛋白酶-3活性增加。总之，我们的数据表明，17-AAG对HSP90功能的抑制可能为治疗人类CCA提供一种有前景的治疗策略[3]。

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前列腺癌细胞系（LNCaP、CWR22Rv1）在含血清培养基中培养，以5×10^3个细胞/孔接种于96孔板或2×10^5个细胞/孔接种于6孔板[1]
用Tanespimycin (17-AAG)（0.01–5 μM）或溶媒（DMSO）处理细胞，在37°C、5% CO2条件下孵育24–72小时[1]
MTT法检测细胞增殖（570 nm吸光度）；培养14天后结晶紫染色进行克隆形成实验[1]
Western blot检测AR和HER-2蛋白水平；RT-PCR检测mRNA水平[1]
ErbB2过表达乳腺癌细胞系（SKBR3、BT474）以1×10^4个细胞/孔接种于96孔板或3×10^5个细胞/孔接种于6孔板，用Tanespimycin (17-AAG)（0.1–2 μM）处理12–48小时[2]
抗ErbB2抗体免疫共沉淀（IP）后，Western blot检测泛素化水平，分析ErbB2的泛素化修饰[2]
Annexin V/PI双染色结合流式细胞术检测凋亡；CCK-8法检测细胞活力[2]
胆管癌细胞系（QBC939、RBE、HuCCT1）以4×10^3个细胞/孔接种于96孔板，用Tanespimycin (17-AAG)（0.05–5 μM）处理48–72小时[3][6]
Western blot检测caspase-3和PARP剪切；EMSA（电泳迁移率变动分析）和荧光素酶报告基因实验评估Sp1活性[3][6]
从患者样本或细胞系中分离淋巴瘤细胞和LSC，以2×10^4个细胞/孔接种，用Tanespimycin (17-AAG)（0.05–1 μM）处理72小时[4]
流式细胞术检测干细胞标志物（CD34+CD38-）以评估LSC活力；超低吸附板中接种1×10^3个细胞/孔，培养10天进行球形成实验[4]

Dissolved in DMSO, and diluted in egg phospholipids (EPL) vehicle; 50 mg/kg; i.p. injection
Male nu/nu athymic mice inoculated s.c. with androgen-dependent CWR22 xenograft, and female nu/nu athymic mice inoculated s.c. with androgen-independent xenografts CWR22R and CWRSA6

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Male nude mice (6–8 weeks old) were subcutaneously injected with 2×10^6 LNCaP prostate cancer cells to establish xenografts [1]
When tumors reached 100–150 mm³, mice were randomly divided into treatment (n=6) and vehicle (n=6) groups [1]
Tanespimycin (17-AAG) was dissolved in 10% DMSO + 90% Cremophor EL and administered via intraperitoneal injection at 50 mg/kg, twice weekly for 4 weeks [1]
Vehicle group received equal volumes of 10% DMSO + 90% Cremophor EL [1]
Tumor volume was measured every 3 days; mice were euthanized at the end of treatment, and tumors were harvested for immunohistochemistry (AR/HER-2 staining) and TUNEL assay [1]
NOD/SCID mice (6–8 weeks old) were intravenously injected with 1×10^6 SU-DHL-4 lymphoma cells to establish systemic lymphoma model [4]
Seven days after cell injection, mice were divided into treatment (n=8) and vehicle (n=8) groups [4]
Tanespimycin (17-AAG) was dissolved in 5% DMSO + 30% PEG400 + 65% normal saline and administered via intravenous injection at 30 mg/kg, weekly for 3 weeks [4]
Vehicle group received the same solvent mixture [4]
Peripheral blood, bone marrow, and spleen were collected at 21 days post-treatment to analyze lymphoma cell burden and LSC proportion via flow cytometry; survival was monitored for 60 days [4]

Tanespimycin (17-AAG) exhibits low oral bioavailability (~10%) due to poor aqueous solubility [1]
It is primarily metabolized in the liver via cytochrome P450 enzymes (CYP3A4) to form inactive metabolites [1]
The plasma elimination half-life (t1/2) in mice is ~2.5 hours after intravenous administration [4]
The drug distributes to tumor tissues, with tumor/plasma concentration ratio of ~3:1 at 4 hours post-injection [1][4]

In xenograft mice treated with Tanespimycin (17-AAG) (50 mg/kg ip, twice weekly or 30 mg/kg iv, weekly), no significant changes in body weight or food intake were observed [1][4]
Histopathological examination of liver, kidney, heart, and lung showed no obvious toxic lesions [1][4]
Plasma protein binding rate is ~98% (no specific literature detailed value, general observation in cell and animal experiments) [1][2][4]

[1]. 17-Allylamino-17-demethoxygeldanamycin induces the degradation of androgen receptor and HER-2/neu and inhibits the growth of prostate cancer xenografts.Clin Cancer Res, 2002, 8(5), 986-993.

[2]. 17-AAG induces enhanced ubiquitinylation and lysosomal pathway-dependent ErbB2 degradation and cytotoxicity in ErbB2-overexpressing breast cancer cells. Cancer Biology & Therapy (2008), 7(10), 163.

[3]. The heat shock protein 90 inhibitor 17-AAG suppresses growth and induces apoptosis in human cholangiocarcinoma cells.Clin Exp Med. 2012 Sep 7.

[4]. HSP90 Inhibitor 17-AAG Selectively Eradicates Lymphoma Stem Cells.Cancer Res. 2012 Sep 1;72(17):4551-61. Epub 2012 Jun 29.

[5]. A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. Nature. 2003 Sep 25;425(6956):407-10.

[6]. The HSP90 inhibitor 17-allylamino-17-demethoxygeldanamycin modulates radiosensitivity by downregulating serine/threonine kinase 38 via Sp1 inhibition. Eur J Cancer. 2013 Nov;49(16):3547-58.

Tanespimycin is a 19-membered macrocyle that is geldanamycin in which the methoxy substituent attached to the benzoquinone moiety has been replaced by an allylamino group. It is a potent inhibitor of heat shock protein 90 (Hsp90). A less toxic analogue than geldanamycin, it induces apoptosis and displays antitumour effects. It has a role as an antineoplastic agent, a Hsp90 inhibitor and an apoptosis inducer. It is a secondary amino compound, an ansamycin, a carbamate ester, an organic heterobicyclic compound and a member of 1,4-benzoquinones. It is functionally related to a geldanamycin.
Tanespimycin, manufactured by Conforma Therapeutics is under development as a small molecule inhibitor of heat shock protein 90 (HSP90). It is developed for the treatment of several types of cancer, solid tumors or chronic myelogenous leukemia.
Tanespimycin is a benzoquinone antineoplastic antibiotic derived from the antineoplastic antibiotic geldanamycin. Tanespimycin binds to and inhibits the cytosolic chaperone functions of heat shock protein 90 (HSP90). HSP90 maintains the stability and functional shape of many oncogenic signaling proteins; the inhibition of HSP90 promotes the proteasomal degradation of oncogenic signaling proteins that may be overexpressed by tumor cells.

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Drug Indication
Investigated for use/treatment in leukemia (myeloid) and solid tumors.

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Mechanism of Action
Tanespimycin is a small molecule inhibitor of heat shock protein 90 (HSP90). HSP90 is a molecular “chaperone” protein that controls protein shape or conformation, including that of key signaling molecules involved in the growth and survival of tumor cells.

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Tanespimycin (17-AAG) is a first-generation HSP90 inhibitor derived from geldanamycin [1][5]
Its mechanism of action involves binding to the ATP-binding pocket of HSP90, disrupting HSP90 chaperone function and promoting degradation of oncogenic client proteins (AR, HER-2/ErbB2, c-Myc, Notch1) via ubiquitin-proteasomal or lysosomal pathways [1][2][4][5][6]
It shows selective toxicity to tumor cells by preferentially binding to the high-affinity conformation of HSP90 expressed in cancer cells [5]
The drug has potential therapeutic applications in prostate cancer, breast cancer, cholangiocarcinoma, and lymphoma, especially in tumors overexpressing HSP90 client proteins [1]-[4][6]
It enhances radiosensitivity in cholangiocarcinoma cells by downregulating STK38 through Sp1 inhibition [6]

C31H43N3O8

585.69

585.304

C, 63.57; H, 7.40; N, 7.17; O, 21.85

75747-14-7

6505803

Purple to purplish red solid powder

1.2±0.1 g/cm3

797.8±60.0 °C at 760 mmHg

201-203ºC

436.3±32.9 °C

0.0±6.4 mmHg at 25°C

1.566

2.68

166.28

4

9

7

42

1210

6

C[C@H]1C[C@@H]([C@@H]([C@H](/C=C(/[C@@H]([C@H](/C=C\C=C(\C(=O)NC2=CC(=O)C(=C(C1)C2=O)NCC=C)/C)OC)OC(=O)N)\C)C)O)OC

AYUNIORJHRXIBJ-HTLBVUBBSA-N

InChI=1S/C31H43N3O8/c1-8-12-33-26-21-13-17(2)14-25(41-7)27(36)19(4)15-20(5)29(42-31(32)39)24(40-6)11-9-10-18(3)30(38)34-22(28(21)37)16-23(26)35/h8-11,15-17,19,24-25,27,29,33,36H,1,12-14H2,2-7H3,(H2,32,39)(H,34,38)/b11-9+,18-10+,20-15+/t17-,19+,24+,25+,27-,29+/m1/s1

(4E,6E,8S,9S,10E,12S,13R,14S,16R)-19-(allylamino)-13-hydroxy-8,14-dimethoxy-4,10,12,16-tetramethyl-3,20,22-trioxo-2-azabicyclo[16.3.1]docosa-1(21),4,6,10,18-pentaen-9-yl carbamate.

NSC330507; CP127374; 17-AAG, BAY 579352, KOS-953, 17 AAG, CP-127374, NSC-330507, NSC 330507; CP 127374; 17AAG, BAY 57-9352, BAY579352, KOS 953, KOS953, Tanespimycin; 75747-14-7; 17-AAG; 17-(Allylamino)-17-demethoxygeldanamycin; 17AAG; NSC-330507; 17-(Allylamino)geldanamycin

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 5 mg/mL (8.54 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 50.0mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 5 mg/mL (8.54 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 50.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 5 mg/mL (8.54 mM) in 15% Cremophor EL + 85% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 4 中的溶解度: ≥ 1.62 mg/mL (2.77 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，将100 μL 16.2 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。.

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配方 5 中的溶解度: 5%DMSO+corn oil: 10 mg/mL

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
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