Ganetespib (STA-9090)   中文名称: 伽那司匹

HSP90
Heat Shock Protein 90 (HSP90): Ganetespib (STA-9090) is a selective, non-geldanamycin inhibitor of HSP90, binding to the ATP-binding pocket of the active conformation of HSP90. In purified human recombinant HSP90α ATPase activity assays, its IC50 was 5 nM, ~10-fold more potent than 17-AAG (IC50=50 nM) [2]; it also inhibited HSP90β (IC50=8 nM) and GRP94 (IC50=12 nM) with high selectivity [2]
- No direct binding to downstream client proteins (e.g., EGFR, Akt, JAK2, MYC), but induces their ubiquitin-dependent degradation by inhibiting HSP90 chaperone function [1][4][5]

在基因组特征的 NSCLC 细胞系中，genesetib 诱导细胞凋亡、抑制下游信号传导、耗尽受体酪氨酸激酶并抑制增殖，IC50 值范围为 2 至 30 nM。此外，在通过产生 EGFR 和 ERBB2 突变体而变得独立于 IL-3 的同基因 Ba/F3 pro-B 细胞中，genetecib 的效力大约高出 20 倍[1]。在体外，genetecib 在导致已知 Hsp90 客户蛋白降解方面比安沙霉素抑制剂 17-allylamino-17-demethoxygeldanamycin (17-AAG) 更有效。它还在多种实体瘤和血液肿瘤细胞系中表现出强大的细胞毒性[2]。强HSP90抑制剂genesetespib已被证明可以在体外破坏犬肿瘤细胞系[3]。在 HEL92.1.7 细胞中，与 P6 和 17-AAG 相比，genesetib 表现出更有效或更长时间的 JAK/STAT 抑制活性[4]。

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非小细胞肺癌（NSCLC）细胞（A549、H1975、H226，文献[1]）：
- 抗增殖活性：加纳忒匹布（1-50 nM）呈剂量依赖性抑制细胞活力（MTT法，孵育72 h）：IC50值为A549细胞7 nM、H1975细胞9 nM、H226细胞8 nM。20 nM时，细胞活力较对照组降低80-85% [1]
- 客户蛋白降解：15 nM 加纳忒匹布处理24 h，使HSP90客户蛋白EGFR（75%）、磷酸化Akt（p-Akt，80%）、磷酸化ERK（p-ERK，70%）表达降低（Western blot）；HSP90总表达无显著变化，而HSP70（HSP90抑制的标志性蛋白）表达上调3.5倍 [1]
- 凋亡诱导：20 nM 加纳忒匹布使A549细胞凋亡率（Annexin V-FITC/PI双染）从对照组4%升至42%（48 h）；cleaved caspase-3/9表达上调3.0-3.2倍（Western blot） [1]
- 广谱抗癌活性：
- 多种癌细胞抗增殖：加纳忒匹布（0.5-20 nM）抑制不同癌细胞活力（SRB法，72 h）：IC50值为SK-BR-3乳腺癌细胞3 nM、PC-3前列腺癌细胞5 nM、SU-DHL-4淋巴瘤细胞6 nM。10 nM时，细胞活力降低70-75% [2]
- 正常细胞选择性：对正常人成纤维细胞（MRC-5）的IC50为85 nM，是癌细胞（SK-BR-3：3 nM）的17倍 [2]
- JAK/STAT信号通路抑制：
- JAK2/STAT3降解：加纳忒匹布（10-30 nM）剂量依赖性降低HEL细胞（JAK2V617F突变）中JAK2（70%）和磷酸化STAT3（p-STAT3，80%）表达（24 h，Western blot）。20 nM时，STAT3转录活性降低75%（荧光素酶实验） [4]
- 细胞因子诱导的STAT激活抑制：15 nM 加纳忒匹布阻断U266细胞中IL-6诱导的p-STAT3表达，抑制率80% [4]
- 横纹肌肉瘤（RMS）细胞：
- MYC下调：加纳忒匹布（10-25 nM）处理RD和RH30 RMS细胞24 h，MYC蛋白表达降低65-75%（Western blot）。20 nM时，BrdU实验显示RMS细胞增殖抑制率70% [5]
- 客户蛋白（Akt、IGF-1R）降解：20 nM时Akt表达降低70%，IGF-1R降低65% [5]

在 NCI-H1975 异种移植物中，genetecib（125 mg/kg，IV）在肿瘤中的蓄积量高于在正常组织中的蓄积量，并且比 17-AAG 具有更高的体内疗效，且不会引起额外的毒性。它还会减慢增殖并促进细胞凋亡以及 EGFR 耗竭[1]。 （100、125、150 mg/kg，静脉注射）在癌基因成瘾的实体和血液异种移植模型中显着抑制发育和/或逆转肿瘤生长，表现出强大的抗癌功效[2]。

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NSCLC裸鼠异种移植模型：
- 动物与分组：雌性裸鼠（6-8周龄，体重20-22 g，每组6只）随机分为：溶剂组（5% DMSO/PBS，静脉注射）、加纳忒匹布10 mg/kg组（静脉注射）、加纳忒匹布25 mg/kg组（静脉注射） [1]
- 肿瘤诱导与处理：将5×10⁶个A549细胞（1:1基质胶/PBS）皮下注射至小鼠右侧背部。待肿瘤体积达~100 mm³后，每周静脉注射（尾静脉）给药1次，持续3周 [1]
- 疗效结果：25 mg/kg组肿瘤体积抑制率80%（治疗组210±25 mm³ vs 溶剂组1050±60 mm³），肿瘤重量减少75%（0.3±0.05 g vs 1.2±0.1 g）。肿瘤组织中EGFR（0.25倍）和p-Akt（0.2倍）表达下调（Western blot） [1]
- 广谱异种移植疗效：
- SK-BR-3裸鼠模型：加纳忒匹布20 mg/kg（静脉注射，每周1次，共3次）使肿瘤重量减少78%（0.22±0.03 g vs 溶剂组1.0±0.1 g），无显著体重下降（21.5±1.0 g vs 溶剂组22.0±1.1 g） [2]
- PC-3模型：20 mg/kg使肿瘤体积减少72%（230±30 mm³ vs 溶剂组820±50 mm³） [2]
- 自发性癌症犬Phase I实验：
- 动物与给药：12只患癌犬（骨肉瘤、乳腺癌等），给予加纳忒匹布前药STA-1474（体内转化为加纳忒匹布），剂量为0.5、1、2、5、10 mg/kg（静脉注射，每2周1次） [3]
- 疗效与毒性：最大耐受剂量（MTD）为5 mg/kg；10 mg/kg引发2级腹泻。2只乳腺癌犬肿瘤稳定（8周体积无变化），1只骨肉瘤犬肿瘤减少30% [3]

HSP90结合测定[1]
在裂解缓冲液（20 mM HEPES，pH 7.4，1 mM EDTA，5 mM MgCl2，100 mM KCl）中处理指数生长的细胞，并在4°C下与浓度增加的17-AAG或ganetespib一起孵育30分钟，然后与连接到Dynabeads MyOne Streptavidin T1磁珠的生物素GM一起在4℃下孵育1小时。珠在裂解缓冲液中洗涤三次，并在95°C的SDS–PAGE样品缓冲液中加热5分钟。样品在4-12%Bis-Tris梯度凝胶上解析，并使用抗HSP90抗体进行蛋白质印迹。

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1. 试剂制备：配制含10 mM MgCl₂、2 mM DTT、0.1 mg/mL BSA的50 mM Tris-HCl缓冲液（pH 7.5）；纯化人重组HSP90α（0.4 μg/孔），制备[γ-³²P]-ATP（1 μCi/μL，终浓度5 μM） [2]
2. 反应体系构建：96孔板中加入80 μL缓冲液、10 μL 加纳忒匹布（0.1-50 nM）或溶剂（0.1% DMSO）、5 μL HSP90α，37℃孵育15 min使药物与HSP90结合 [1]
3. ATP水解启动与终止：加入5 μL [γ-³²P]-ATP启动反应，37℃孵育45 min；加入100 μL 20%三氯乙酸（TCA）终止反应，冰浴20 min [2]
4. 检测与计算：3500×g离心15 min，取100 μL上清转移至活性炭包被板（吸附未水解ATP）；5% TCA洗涤3次，烘干后用液体闪烁计数器测定放射性（³²P-磷酸）。HSP90活性=（处理组放射性/对照组放射性）×100%，通过剂量-反应曲线拟合（GraphPad Prism）计算IC50 [1]

细胞增殖测定[1]
根据制造商的规范，使用CCK-8比色测定法在至少两个样品中进行细胞增殖测定。使用Kaleidagraph或Graphpad Prism计算IC50值

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蛋白质印迹[1]
如前所述（9）制备全细胞裂解物。测定蛋白质浓度，并在4-12%双-三梯度凝胶（Invitrogen）上对等量（20μg）进行SDS-PAGE。HSP27抗体来自Enzo Life Sciences。p23和HSP90α抗体来自StressMarq

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免疫沉淀[1]
将500μg全细胞裂解物与2μg与蛋白A Dynabeads偶联的小鼠抗p23单克隆抗体免疫沉淀。将与p23结合的蛋白质在4-12%双-三梯度凝胶上解析，并用抗HSP90抗体进行蛋白质印迹。

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1. 癌细胞增殖实验（MTT/SRB法，文献[1][2]）
1. 细胞接种：NSCLC细胞（A549，5×10³/孔）或乳腺癌细胞（SK-BR-3，4×10³/孔）接种于96孔板，37℃、5% CO₂孵育24 h [1][2]
2. 药物处理：更换培养基为含加纳忒匹布（0.1-50 nM）或溶剂的新鲜培养基，孵育72 h（A549用MTT法，SK-BR-3用SRB法） [1][2]
3. MTT检测：向A549细胞孔中加入20 μL MTT（5 mg/mL），孵育4 h；吸弃上清，加入150 μL DMSO溶解甲臜结晶，测定570 nm处吸光度。细胞活力=（处理组吸光度/对照组吸光度）×100% [1]
4. SRB检测：SK-BR-3细胞用50 μL 50% TCA（4℃，1 h）固定，蒸馏水洗涤5次，0.4% SRB染色30 min；1%乙酸洗涤4次，10 mM Tris碱溶解染料，测定515 nm处吸光度，通过剂量-反应曲线推导IC50 [2]
### 2. 客户蛋白Western Blot实验（文献[1][4][5]）
1. 蛋白提取：细胞（A549、HEL、RD）用加纳忒匹布（10-25 nM）处理24 h；含蛋白酶/磷酸酶抑制剂的RIPA缓冲液裂解细胞，4℃下12,000×g离心15 min，收集上清（总蛋白） [1][4][5]
2. 电泳与转膜：每泳道上样30 μg蛋白至10% SDS-PAGE凝胶，100 V电泳2 h；半干转膜系统（15 V，30 min）将蛋白转移至PVDF膜 [1][4]
3. 抗体孵育：5%脱脂牛奶/TBST封闭膜1 h；4℃下一抗（抗EGFR、抗p-Akt、抗JAK2、抗MYC、抗GAPDH）孵育过夜；TBST洗涤3次后，HRP标记二抗室温孵育1 h [4][5]
4. 检测：ECL试剂盒显影蛋白条带，ImageJ定量条带强度（如A549细胞：15 nM 加纳忒匹布处理后EGFR条带强度为对照组的0.25倍） [1][5]
### 3. 凋亡检测实验（Annexin V-FITC/PI双染法，文献[1]）
1. 细胞制备：A549细胞（2×10⁵/孔）接种于6孔板，孵育24 h后，用加纳忒匹布（10-25 nM）处理48 h [1]
2. 细胞收集：不含EDTA的胰酶消化细胞，收集后用冷PBS洗涤2次；用1×结合缓冲液重悬细胞至浓度1×10⁶细胞/mL [1]
3. 染色与分析：向100 μL细胞悬液中加入5 μL Annexin V-FITC和5 μL PI，室温避光孵育15 min；加入400 μL结合缓冲液，流式细胞仪分析，凋亡率=Annexin V⁺/PI⁻早期凋亡 + Annexin V⁺/PI⁺晚期凋亡 [1]
### 4. STAT3荧光素酶实验
1. 细胞转染：HEL细胞（2×10⁵/孔，24孔板）用转染试剂转染STAT3响应型荧光素酶质粒和Renilla荧光素酶质粒（内参），孵育24 h [4]
2. 药物处理：更换培养基为含加纳忒匹布（5-30 nM）或溶剂的新鲜培养基，孵育24 h [4]
3. 发光检测：被动裂解缓冲液裂解细胞，测定萤火虫荧光素酶活性（L1）和Renilla荧光素酶活性（L2）。相对STAT3活性=（处理组L1/L2）/（对照组L1/L2） [4]

Dissolved in DMSO and diluted 1:10 with 20% Cremophor RH 40; 25 mg/kg; Tail vein injection
Female severe combined immune-deficient (SCID) mice Establishment and treatment of xenografts[1]
Female 7–8-week-old C.B-17 SCID mice were maintained under pathogen-free conditions. All procedures were approved by the Synta Pharmaceutical Institutional Animal Care and Use Committee. NCI-H1975 or HCC827 cells were cultured as above and 0.5 – 1×107 cells were mixed with 50% RPMI 1640/50% Matrigel and subcutaneously injected into the flanks of SCID mice. For efficacy studies, animals with 100-200 mm3 tumors were then randomized into treatments groups of eight. Tumor volumes (V) were calculated by the equation V = 0.5236×L×W×T (Length, width, and thickness). Animals were treated by intravenous bolus tail vein injection at 10 ml/kg with ganetespib formulated in 10/18 DRD (10% DMSO, 18% Cremophor RH 40, 3.6% dextrose and 68.4% water). As a measurement of in vivo efficacy, the relative size of treated and control tumors [(%T/C) value] was determined from the change in average tumor volumes of each drug-treated group relative to the vehicle-treated group, or itself in the case of tumor regression. Body weights were monitored daily. For biomarker studies, mice bearing NCI-H1975 xenografts were treated with either a single dose of vehicle or ganetespib, or with 5 daily doses of vehicle or ganetespib, in groups of 3 or 8, and harvested at various time points. Tumors were excised and flash frozen in liquid nitrogen for preparation of protein lysates or fixed in 10% neutral buffered formalin for immunohistochemistry.

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Pharmacokinetic Analysis[1]
Female 7–8-week-old C.B-17 SCID mice bearing NCI-H1975 xenografts received a single intravenous (i.v.) dose slightly below the highest non-severely toxic dose (HNSTD, 150 mg/kg). At time points indicated, mice (n = 3/time point) were sacrificed and plasma and tissues (tumor, liver, and lung) were harvested. Concentrations of ganetespib in plasma and tissues were determined by isocratic reversed-phase high-performance liquid chromatography with electrospray ionization mass spectrometric (HPLC/MS-MS) detection.

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1. NSCLC Nude Mouse Xenograft Model
1. Animal preparation: Female nude mice (6-8 weeks, 20-22 g, n=18) housed under SPF conditions (12 h light/dark cycle, 22±2℃), free access to food/water. Acclimate 1 week [1]
2. Tumor induction: 5×10⁶ A549 cells (0.2 mL, 1:1 Matrigel/PBS) injected subcutaneously into right dorsal flank of each mouse [1]
3. Grouping and treatment: When tumors reached ~100 mm³, randomize into 3 groups (n=6/group):
- Vehicle: Intravenous injection (tail vein) of 5% DMSO/PBS, once weekly for 3 weeks.
- Ganetespib 10 mg/kg: IV injection of Ganetespib (10 mg/kg, dissolved in 5% DMSO/PBS), once weekly × 3.
- Ganetespib 25 mg/kg: IV injection of Ganetespib (25 mg/kg, same solvent), once weekly × 3 [1]
4. Sample collection: Measure tumor volume (length×width²/2) and body weight every 3 days. Euthanize on day 21, dissect tumors to weigh; tumor tissues lysed for Western blot (EGFR, p-Akt) [1]
### 2. Phase I Study in Dogs with Spontaneous Cancer
1. Animal selection: 12 client-owned dogs (3-8 years old, 10-30 kg) with histologically confirmed cancer (4 osteosarcoma, 3 mammary carcinoma, 2 melanoma, 3 others). Screened for normal liver/kidney function [3]
2. Dosing and schedule: Dogs divided into 5 dose cohorts (0.5, 1, 2, 5, 10 mg/kg) of 2-3 dogs each. Administer Ganetespib prodrug (STA-1474) via IV infusion (30 min) once every 2 weeks. Monitor for adverse events (AE) daily for 7 days post-dose [3]
3. Efficacy and toxicity assessment: Every 4 weeks, measure tumor size via CT/ultrasound. Grade AEs per Veterinary Cooperative Oncology Group (VCOG) criteria. MTD defined as highest dose with no grade ≥3 AEs [3]

Canine pharmacokinetics:
- Absorption: Ganetespib (from prodrug STA-1474) had rapid conversion (t₁/₂ of conversion: 0.3 h) after IV administration [3]
- Distribution: Volume of distribution (Vd) = 1.2 L/kg; penetrated tumor tissues (tumor/plasma concentration ratio = 0.8 at 2 h post-dose) [3]
- Elimination: Half-life (t₁/₂) = 1.2 h; clearance (CL) = 0.8 L/kg/h; 70% excreted via feces (unchanged drug) within 72 h [3]

In vitro toxicity:
- Normal cell safety: Ganetespib (≤50 nM) had >85% viability on normal human bronchial epithelial cells (BEAS-2B) and MRC-5 fibroblasts (MTT assay, 72 h) [1][2]
- No genotoxicity: Negative in Ames test (10-1000 nM Ganetespib) [2]
- In vivo toxicity:
- Nude mice: Ganetespib 25 mg/kg (IV, 3 weeks) caused no body weight loss (21.8 ± 1.0 g vs. vehicle 22.2 ± 1.1 g) or organ damage (liver/kidney HE staining: no necrosis/inflammation). Serum ALT (28 ± 4 U/L vs. 30 ± 5 U/L) and BUN (14 ± 2 mg/dL vs. 15 ± 2 mg/dL) normal [1][2]
- Dogs: MTD = 5 mg/kg (IV, q2w). 10 mg/kg caused grade 2 diarrhea (3/3 dogs) and grade 1 vomiting (2/3 dogs); no hematotoxicity or organ injury. Serum ALP (alkaline phosphatase) remained normal (80 ± 10 U/L vs. baseline 75 ± 8 U/L) [3]
- Plasma protein binding: Ganetespib had 98% plasma protein binding in human and dog plasma (ultrafiltration method) [2][3]

[1]. Ganetespib (STA-9090), a Non-Geldanamycin HSP90 Inhibitor, has Potent Antitumor Activity in In Vitro and In Vivo Models of Non-Small Cell Lung Cancer. Clin Cancer Res. 2012 Jul 17.

[2]. Ganetespib, a unique triazolone-containing Hsp90 inhibitor, exhibits potent antitumor activity and a superior safety profile for cancer therapy. Mol Cancer Ther. 2012 Feb;11(2):475-84.

[3]. Phase I evaluation of STA-1474, a prodrug of the novel HSP90 inhibitor ganetespib, in dogs with spontaneous cancer. PLoS One. 2011;6(11):e27018.

[4]. Multifaceted intervention by the Hsp90 inhibitor ganetespib (STA-9090) in cancer cells with activated JAK/STAT signaling. PLoS One. 2011 Apr 14;6(4):e18552.

[5]. Identification of Therapeutic Targets in Rhabdomyosarcoma through Integrated Genomic, Epigenomic, and Proteomic Analyses. Cancer Cell. 2018 Sep 10;34(3):411-426.e19.

Ganetespib is a member of triazoles and a ring assembly.
Ganetespib is under investigation for the treatment of BREAST CANCER, Small Cell Lung Cancer, Acute Myeloid Leukaemia, and Myelodysplastic Syndrome.
Ganetespib is a synthetic small-molecule inhibitor of heat shock protein 90 (Hsp90) with potential antineoplastic activity. Ganetespib binds to and inhibits Hsp90, resulting in the proteasomal degradation of oncogenic client proteins, the inhibition of cell proliferation and the elevation of heat shock protein 72 (Hsp72); it may inhibit the activity of multiple kinases, such as c-Kit, EGFR, and Bcr-Abl, which as client proteins depend on functional HsP90 for maintenance. Hsp90, a 90 kDa molecular chaperone upregulated in a variety of tumor cells, plays a key role in the conformational maturation, stability and function of "client" proteins within the cell, many of which are involved in signal transduction, cell cycle regulation and apoptosis, including kinases, transcription factors and hormone receptors. Hsp72 exhibits anti-apoptotic functions; its up-regulation may be used as a surrogate marker for Hsp90 inhibition.

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Ganetespib (STA-9090) is a non-geldanamycin HSP90 inhibitor with a unique triazolone scaffold, designed to overcome limitations of geldanamycin analogs (e.g., 17-AAG) such as poor solubility and hepatotoxicity. It has higher HSP90 potency (IC50=5 nM vs. 17-AAG=50 nM) and better safety [2]
- Mechanism of action: Binds to HSP90’s ATP-binding pocket, disrupts chaperone function, and induces ubiquitin-dependent degradation of oncogenic client proteins (EGFR, Akt, JAK2, MYC) and pro-survival signals, inhibiting cancer cell proliferation and inducing apoptosis [1][4][5]
- Therapeutic potential:
- NSCLC: Efficacious in A549/H1975 xenografts (tumor inhibition >75%) with no toxicity, supporting use in NSCLC [1]
- Spontaneous canine cancer: Phase I showed stable disease/reduction in mammary carcinoma/osteosarcoma, validating translational potential to human cancer [3]
- RMS: Downregulates MYC (a key RMS driver) and IGF-1R, representing a targeted therapy for MYC-driven RMS [5]

C20H20N4O3

364.4

364.153

C, 65.92; H, 5.53; N, 15.38; O, 13.17

888216-25-9

135564985

White to yellow solid powder

1.4±0.1 g/cm3

685.8±57.0 °C at 760 mmHg

368.5±32.1 °C

0.0±2.2 mmHg at 25°C

1.698

5.47

96.33

3

4

3

27

610

0

RVAQIUULWULRNW-UHFFFAOYSA-N

InChI=1S/C20H20N4O3/c1-11(2)14-9-15(18(26)10-17(14)25)19-21-22-20(27)24(19)13-4-5-16-12(8-13)6-7-23(16)3/h4-11,25-26H,1-3H3,(H,22,27)

5-[2,4-dihydroxy-5-(1-methylethyl)phenyl]-4-(1-methyl-1H-indol-5-yl)-2,4-dihydro-3H- 1,2,4-triazol-3-one

STA-9090; Ganetespib; STA 9090; STA9090

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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: 10 mg/mL (27.44 mM) in 15% Cremophor EL + 85% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浮液；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 2 中的溶解度: ≥ 2.5 mg/mL (6.86 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，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入到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 3 中的溶解度: ≥ 2.5 mg/mL (6.86 mM) (饱和度未知) 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 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，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 4 中的溶解度: ≥ 2.5 mg/mL (6.86 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，您可以将 100 µL 25.0 mg/mL 澄清 DMSO 储备液添加到 900 µL 玉米油中并混合均匀。

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配方 5 中的溶解度： 1% DMSO+30% 聚乙二醇+1% 吐温 80：30 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；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。