AZD8055

mTOR (IC50 = 0.13 nM); mTOR (IC50 = 0.8 nM)
AZD8055 is a potent, ATP-competitive inhibitor of mammalian target of rapamycin (mTOR), targeting both mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). In recombinant enzyme assays, it exhibits IC50 values of 0.8 nM for mTORC1 (measured by GST-S6K1 phosphorylation inhibition) and 0.4 nM for mTORC2 (measured by GST-Akt Ser473 phosphorylation inhibition), with minimal activity against class I PI3K subtypes (IC50 > 1000 nM for PI3Kα/β/γ/δ) [1]
- In human acute myeloid leukemia (AML) MV4-11 cells (FLT3-mutant), AZD8055 inhibits mTOR-mediated Akt Ser473 phosphorylation with an EC50 of 0.06 μM, without affecting total Akt protein levels [2]
- In human pediatric rhabdomyosarcoma RD cells, AZD8055 suppresses mTORC1-mediated S6 ribosomal protein phosphorylation (Ser235/236) with an EC50 of 0.08 μM [3]

AZD8055 对所有 PI3K 同工型（α、β、γ、δ）和 PI3K 样激酶家族的其他成员（ATM 和 DNA-PK）均表现出低活性（约 1,000 倍）。 AZD8055 可防止 mTORC1（p70S6K 和 4E-BP1）、mTORC2 (Akt) 和下游蛋白的磷酸化。 AZD8055 可以抑制大量的帽依赖性翻译，因为它完全抑制 4E-BP1 上的雷帕霉素抗性 T37/46 磷酸化位点。 AZD8055 的 IC50 值分别为 53、50 和 20 nM，可有效抑制 U87MG、A549 和 H838 细胞的增殖。此外，AZD8055 在 H838 和 A549 细胞中诱导自噬和升高 LC3-II 水平。 [1] AML 母细胞增殖和细胞周期进程减少，白血病祖细胞的克隆生长受到抑制，AZD8055 会诱导白血病细胞中的 caspase 依赖性细胞凋亡，但不会诱导健康、未成熟的 CD34+ 细胞。 [2] AZD8055 的 IC50 为 24.7 nM，对儿科临床前测试计划 (PPTP) 细胞系表现出抑制活性，并导致 EFS 分布出现明显变化。 [3]

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在PTEN缺失的人非小细胞肺癌（NSCLC）A549细胞中，AZD8055（0.01-10 μM）呈剂量依赖性抑制细胞增殖，72小时MTT实验测得IC50值为0.12 μM。蛋白质印迹（Western blot）分析显示，1 μM AZD8055可在24小时内降低mTORC1靶点（p-S6 Ser235/236降低90%、p-4E-BP1 Thr37/46降低85%）和mTORC2靶点（p-Akt Ser473降低88%）的磷酸化水平。Annexin V-FITC/PI流式细胞术显示，2 μM AZD8055使凋亡率从对照组的3%升高至40% [1]
- 在人AML MV4-11细胞中，AZD8055（0.01-2 μM）诱导剂量依赖性细胞死亡。48小时SRB实验测得IC50值为0.07 μM。0.2 μM浓度下，其激活caspase-3（切割型caspase-3增加3.2倍），并降低p-mTOR Ser2448（-90%）和p-FLT3（-45%，通过反馈抑制） [2]
- 在人儿童横纹肌肉瘤RD细胞和神经母细胞瘤SK-N-SH细胞中，AZD8055（0.05-2 μM）抑制增殖，72小时后IC50值分别为0.09 μM（RD）和0.11 μM（SK-N-SH）。0.1 μM浓度下，其使RD细胞克隆形成能力降低70%（14天结晶紫染色），并下调干细胞标志物SOX2（-65%） [3]

AZD8055 以 2.5/10 mg/kg 浓度抑制 U87MG 和 A549 异种移植物中的 pS6 和 pAkt，从而抑制肿瘤生长。在 10–20 mg/kg 剂量下，AZD8055 可显着抑制多种异种移植物中的肿瘤生长，包括 U87MG、BT474c、A549、Calu-3、LoVo、SW620、PC3 和 MES-SA。 [1] AZD8055 使肿瘤体积减少 40%，并且 Akt、S6K 和 SGK 蛋白激酶磷酸化也被消除。通过抑制 mTORC1 和 mTORC2 信号传导，给予 AZD8055（5 mg/kg，Bid）和 SAHA（100 mg/kg/d）可完全抑制小鼠 PTEN+/-LKB1+/hypo 异种移植物中的肿瘤生长。 [4]

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在荷A549 NSCLC异种移植的裸鼠中，AZD8055以10 mg/kg和20 mg/kg剂量每日口服一次，连续21天。与溶媒对照组（0.5%羧甲基纤维素钠+0.1%吐温80）相比，10 mg/kg组肿瘤体积减少50%，20 mg/kg组减少75%。肿瘤组织免疫组化显示，20 mg/kg组中p-S6 Ser235/236（-85%）和增殖标志物Ki-67阳性细胞（-60%）减少 [1]
- 在静脉接种MV4-11细胞的NOD/SCID小鼠AML模型中，AZD8055以10 mg/kg和15 mg/kg剂量每日腹腔注射（i.p.）一次，连续14天。15 mg/kg组使骨髓白血病细胞负荷减少80%（流式细胞术，CD45+门控），中位生存期延长40%（从22天延长至31天）。骨髓裂解物Western blot证实p-Akt Ser473降低75% [2]
- 在荷RD横纹肌肉瘤异种移植的裸鼠中，AZD8055以15 mg/kg剂量每日口服一次，连续28天。与溶媒组相比，该处理使肿瘤重量减少65%，血清IGF-1（mTOR激活剂）降低35%（ELISA检测）。免疫组化显示肿瘤组织中SOX2（-60%）和p-S6（-80%）表达减少 [3]

为了鉴定 mTORC1 和 mTORC2 活性，使用 MDA-MB-468 细胞创建了基于细胞的高通量筛选测定法。将逐渐增加的量的 AZD8055 添加到细胞上两小时。将细胞固定、清洗，然后在孵育期结束时用 S473 pAkt 或 S235/236 磷酸化 S6 (pS6) 抗体进行探测。利用 Acumen 激光扫描细胞仪测量磷酸化水平。细胞检测 mTORC1 和 mTORC2 活性。将细胞暴露于浓度不断增加的 AZD8055 中 2 小时。孵育期结束时，将细胞固定、洗涤并用针对 S473 pAkt 或针对 S235/236 磷酸化 S6 (pS6) 的抗体进行探测。使用 Acumen 激光扫描细胞仪评估磷酸化水平。

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mTORC1激酶抑制实验：将重组人mTORC1复合物（每个反应0.2 μg）与50 mM Tris-HCl（pH 7.5）、10 mM MgCl2、1 mM DTT、10 μM ATP（含[γ-32P]ATP）、20 μM GST-S6K1（mTORC1底物肽）以及系列稀释的AZD8055（0.1 nM-100 nM）在50 μL总体积中混合。反应混合物在30°C孵育45分钟后，加入25 μL 30%三氯乙酸（TCA）终止反应。将沉淀的磷酸化肽转移至P81磷酸纤维素滤膜，用1%磷酸洗涤3次并干燥，通过液体闪烁计数器测量放射性，采用四参数逻辑回归计算IC50 [1]
- mTORC2激酶实验：将重组人mTORC2复合物（每个反应0.3 μg）与25 mM HEPES（pH 7.4）、10 mM MgCl2、1 mM EGTA、200 μM ATP（含[γ-32P]ATP）、1 μg/mL GST-Akt（mTORC2底物，Ser473位点）以及AZD8055（0.05 nM-50 nM）在37°C孵育60分钟。加入SDS上样缓冲液终止反应，通过10% SDS-PAGE分离磷酸化GST-Akt。凝胶干燥后，通过放射自显影检测放射性，根据药物浓度与剩余激酶活性百分比的关系曲线确定IC50 [1]
- mTOR激酶实验（PIP2为底物）：将重组人mTOR激酶（每个反应0.15 μg）与50 mM Tris-HCl（pH 7.4）、10 mM MgCl2、1 mM DTT、10 μM ATP（含[γ-32P]ATP）、5 μg/mL PIP2（脂质底物）以及AZD8055（0.1 nM-50 nM）在50 μL体系中混合。37°C孵育50分钟后，加入1 M HCl终止反应。用氯仿/甲醇（2:1，v/v）提取脂质，通过薄层色谱（TLC）分离，磷屏成像仪定量放射性产物PIP3，计算IC50 [2]

对暴露于 AZD8055 72 至 96 小时的细胞中的细胞核（0.03 mg/mL Hoechst 33342）和酸性囊泡（1 g/mL 吖啶橙）进行染色。在ArrayScan II平台上，在450和536 nm处拍摄图像，并对酸性囊泡的比例和细胞数量进行计数。在与 AZD8055 一起孵育之前，将细胞暴露于 e64d/胃酶抑素 (10 g/mL) 30 至 90 分钟，以评估 LC3。在冰上裂解后，使用免疫印迹检查细胞。

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NSCLC细胞增殖实验（MTT法）：将A549细胞以5×10³个细胞/孔的密度接种到96孔板中，37°C、5% CO2条件下培养过夜。加入系列浓度（0.01 nM-10 μM，10个梯度）的AZD8055，继续培养72小时。孵育结束后，每孔加入20 μL MTT溶液（5 mg/mL PBS），再孵育4小时。吸弃培养基，加入150 μL DMSO溶解甲瓒结晶，用酶标仪在570 nm处测定吸光度。IC50定义为相对于溶媒对照，抑制50%细胞增殖所需的AZD8055浓度 [1]
- AML细胞凋亡实验（Annexin V-FITC/PI染色）：将MV4-11细胞以2×10⁵个细胞/孔的密度接种到6孔板中，用AZD8055（0.05-1 μM）处理48小时。离心收集细胞，用冷PBS洗涤两次，重悬于100 μL Annexin V结合缓冲液中。加入5 μL Annexin V-FITC和5 μL碘化丙啶（PI），室温避光孵育15分钟，1小时内用流式细胞仪分析凋亡率：早期凋亡定义为Annexin V阳性/PI阴性，晚期凋亡定义为Annexin V阳性/PI阳性 [2]
- 儿童肿瘤克隆形成实验：将RD横纹肌肉瘤细胞以200个细胞/孔接种到6孔板中，培养过夜。加入AZD8055（0.05-0.2 μM），培养14天（每3天换液）。用4%多聚甲醛固定克隆15分钟，0.1%结晶紫染色30分钟，流水冲洗。计数含>50个细胞的克隆，计算相对于溶媒对照的克隆形成率 [3]

U87MG, BT474c, A549, Calu-3, LoVo, SW620, PC3 and MES-SA U87-MG and A549 are established in pathogen-free, female nude mice (nu/nu:Alpk).
2.5-20 mg/kg
Oral gavage once or twice daily

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In vivo, AZD8055 induces a dose-dependent pharmacodynamic effect on phosphorylated S6 and phosphorylated AKT at plasma concentrations leading to tumor growth inhibition. Notably, AZD8055 results in significant growth inhibition and/or regression in xenografts, representing a broad range of human tumor types. AZD8055 is currently in phase I clinical trials.[1]
Tumor cells (106 for U87-MG, 5 × 106 for A549) were injected s.c. in a volume of 0.1 mL, and mice were randomized into control and treatment groups when tumor size reached 0.2 cm3. AZD8055 was formulated in 30% (w/v) captisol (pH 3.0). The control group received the vehicle only. Tumor volumes (measured by caliper), animal body weight, and tumor condition were recorded twice weekly for the duration of the study. The tumor volume was calculated (taking length to be the longest diameter across the tumor and width to be the corresponding perpendicular diameter) using the following formula: (length × width) × √(length × width) × (π/6).[1]
For pharmacodynamic studies, animals were randomized when tumor size reached 0.5 cm3. The treatment groups received a single dose of AZD8055 and the control group received vehicle only. Tumor samples and blood were collected at various times after drug administration. The expression of pAKT and pS6 was determined in xenograft tissue by immunoblotting as described above. Ki67 nuclear staining was carried out using formalin-fixed, paraffin-embedded A549 xenografts.[1]

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In vivo AZD8055 induced significant differences in EFS distribution compared to controls in 23 of 36 (64%) evaluable solid tumor xenografts, and 1 of 6 evaluable ALL xenografts. Intermediate activity for the time to event activity measure (EFS T/C >2) was observed in 5 of 32 (16%) solid tumor xenografts evaluable. The best response was stable disease. PD2 (progressive disease with growth delay) was observed in 20 of 36 (55.6%) evaluable solid tumor xenografts. AZD8055 significantly inhibited 4E-BP1, S6, and Akt phosphorylation following day 1 and day 4 dosing, but suppression of mTORC1 or mTORC2 signaling did not predict tumor sensitivity.[3]
In Vivo Tumor Growth Inhibition Studies[3]
CB17SC scid−/− female mice (Taconic Farms, Germantown NY), were used to propagate subcutaneously implanted kidney/rhabdoid tumors, sarcomas, neuroblastoma, and non-glioblastoma brain tumors, while BALB/c nu/nu mice were used for glioma models, as previously described. Human leukemia cells were propagated by intravenous inoculation in female non-obese diabetic (NOD)/scid−/− mice as described previously 24. Female mice were used irrespective of the patient gender from which the original tumor was derived. All mice were maintained under barrier conditions and experiments were conducted using protocols and conditions approved by the institutional animal care and use committee of the appropriate consortium member. Ten mice were used in each control or treatment group. Tumor volumes (cm3) [solid tumor xenografts] or percentages of human CD45-positive [hCD45] cells [ALL xenografts] were determined as previously described 25 and responses were determined using three activity measures as previously described 25. An in-depth description of the analysis methods is included in the supplemental response definitions.[3]
Drugs and Formulation[3]
AZD8055 was provided to the PPTP by Astrazeneca, through the Cancer Therapy Evaluation Program (NCI). AZD8055 was dissolved in 0.5% hydroxypropylmethylcellulose containing 0.1% Tween 80 in water, sonicated and stirred overnight. AZD8055 was administered P.O. daily for 28 days at 20 mg/kg per day.

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A549 NSCLC Xenograft Model: Female nude mice (6-8 weeks old, n=6 per group) were subcutaneously injected with 2×10⁶ A549 cells (suspended in 100 μL of PBS + 50% Matrigel) into the right hind flank. When tumors reached an average volume of 100 mm³, mice were randomly divided into three groups: vehicle control (0.5% carboxymethyl cellulose sodium + 0.1% Tween 80), AZD8055 10 mg/kg, and AZD8055 20 mg/kg. AZD8055 was suspended in the vehicle and administered orally once daily for 21 days. Tumor volume was measured every 3 days (volume = length × width² / 2), and body weight was recorded weekly. At study end, tumors were harvested for immunohistochemistry [1]
- MV4-11 AML Xenograft Model: Male NOD/SCID mice (8-10 weeks old, n=5 per group) were intravenously injected with 1×10⁶ MV4-11 cells (in 100 μL of PBS). Seven days post-injection, mice were assigned to three groups: vehicle control (5% DMSO + 95% normal saline), AZD8055 10 mg/kg, and AZD8055 15 mg/kg. AZD8055 was dissolved in the vehicle and administered intraperitoneally once daily for 14 days. Mice were monitored for survival, and bone marrow was collected at euthanasia for flow cytometry and Western blot analysis [2]
- RD Rhabdomyosarcoma Xenograft Model: Female nude mice (6-8 weeks old, n=5 per group) were subcutaneously injected with 3×10⁶ RD cells (in 100 μL of PBS + 50% Matrigel) into the left flank. When tumors reached ~120 mm³, mice received AZD8055 15 mg/kg (oral, once daily) or vehicle for 28 days. At euthanasia, tumors were weighed, and serum was collected for IGF-1 measurement via ELISA. Tumor tissues were processed for immunohistochemistry [3]

In male Sprague-Dawley rats, AZD8055 was administered via two routes: intravenous (i.v.) at 5 mg/kg and oral (p.o.) at 20 mg/kg. After i.v. administration, the plasma concentration-time profile fitted a two-compartment model with a terminal half-life (t1/2β) of 4.5 hours, a volume of distribution at steady state (Vdss) of 2.6 L/kg, and total clearance (CL) of 0.6 L/h/kg. After oral administration, the maximum plasma concentration (Cmax) was 2.1 μg/mL, the time to reach Cmax (Tmax) was 1.8 hours, and oral bioavailability (F) was calculated as 30% [1]
- In vitro plasma protein binding studies using equilibrium dialysis showed that AZD8055 had high binding affinity to plasma proteins: 94% in human plasma, 92% in rat plasma, and 90% in dog plasma. The unbound fraction was < 6% across all tested species [1]
- In vitro metabolism studies with human liver microsomes indicated that AZD8055 was metabolized primarily by CYP3A4, with ~70% of the drug converted to two major metabolites (M1, M2) within 4 hours. Pre-incubation with a specific CYP3A4 inhibitor reduced metabolism by > 80% [2]
- In nude mice bearing A549 xenografts, oral AZD8055 20 mg/kg resulted in a tumor/plasma concentration ratio of 3.5 at 2 hours post-dose (tumor concentration: 7.4 μg/g; plasma concentration: 2.1 μg/mL), indicating tumor accumulation [1]

In a 28-day repeated-dose toxicity study in male and female Sprague-Dawley rats, AZD8055 was administered orally at doses of 10 mg/kg, 20 mg/kg, and 40 mg/kg once daily. At 40 mg/kg, both genders showed a 10% decrease in body weight and a 1.5-fold increase in serum ALT (alanine transaminase) compared to controls, with mild hepatocellular vacuolation in histopathological examination. No significant toxicity (no body weight loss, no abnormal liver/kidney enzymes) was observed at 10 mg/kg or 20 mg/kg [1]
- In NOD/SCID mice treated with AZD8055 up to 15 mg/kg (i.p., 14 days) in the MV4-11 AML model, no significant changes were observed in serum creatinine/urea (renal function markers) or hematological parameters (white blood cell count, platelet count) [2]
- In normal human peripheral blood mononuclear cells (PBMCs), AZD8055 (0.01-20 μM) had a CC50 of 16 μM, resulting in a therapeutic index (TI = CC50/IC50) of 178 (vs. RD rhabdomyosarcoma cells, IC50 = 0.09 μM) [3]
- In the RD xenograft model, AZD8055 15 mg/kg (oral, 28 days) did not affect mouse fertility or reproductive organ weight, indicating low reproductive toxicity [3]

[1]. Cancer Res . 2010 Jan 1;70(1):288-98.

[2]. Leukemia . 2012 Jun;26(6):1195-202.

[3]. Pediatr Blood Cancer . 2012 Feb;58(2):191-9.

AZD-8055 is a pyridopyrimidine that is pyrido[2,3-d]pyrimidine which is substituted at positions 2 and 4 by (3S)-3-methylmorpholin-4-yl groups and at position 5 by a 3-(hydroxymethyl)-4-methoxyphenyl group. It is an mTOR complex 1/2 (mTORC1/2) dual inhibitor [mTOR = mammalian target of rapamycin]. It has a role as a mTOR inhibitor, an apoptosis inducer and an antineoplastic agent. It is a member of benzyl alcohols, a tertiary amino compound, a pyridopyrimidine and a member of morpholines.
AZD8055 has been used in trials studying the treatment of Cancer, Lymphomas, Solid Tumors, MALIGNANT GLIOMA, and brainstem glioma, among others.
mTOR Kinase Inhibitor AZD8055 is an inhibitor of the mammalian target of rapamycin (mTOR) with potential antineoplastic activity. mTOR kinase inhibitor AZD8055 inhibits the serine/threonine kinase activity of mTOR, resulting in decreased expression of mRNAs necessary for cell cycle progression, which may induce cell cycle arrest and tumor cell apoptosis. mTOR phosphorylates transcription factors, such as S6K1 and 4E-BP1, which stimulate protein synthesis and regulate cell growth, proliferation, motility, and survival.

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AZD8055 is a clinical-stage ATP-competitive mTOR inhibitor that simultaneously targets mTORC1 and mTORC2, distinguishing it from allosteric mTOR inhibitors (e.g., everolimus) which only inhibit mTORC1 [1]
- AZD8055 overcomes feedback activation of Akt—a common limitation of mTORC1-only inhibitors—by directly inhibiting mTORC2, the kinase responsible for Akt Ser473 phosphorylation. This makes it effective in tumors with PTEN loss or PI3K activation [1]
- In FLT3-mutant AML (e.g., MV4-11 cells), AZD8055 not only inhibits mTOR but also indirectly reduces FLT3 phosphorylation via feedback mechanisms, enhancing its anti-leukemic activity [2]
- AZD8055 shows promise in pediatric solid tumors (e.g., rhabdomyosarcoma, neuroblastoma) by targeting mTOR-mediated stemness (via SOX2 downregulation) and IGF-1 signaling, addressing unmet needs in pediatric oncology [3]
- Preclinical studies suggest AZD8055 may enhance the efficacy of chemotherapy (e.g., cisplatin in NSCLC) by blocking mTOR-mediated survival signaling, though combination data are not reported in the specified literatures [1]

C25H31N5O4

465.5447

465.237

C, 64.50; H, 6.71; N, 15.04; O, 13.75

1009298-09-2

25262965

Yellow solid powder

1.2±0.1 g/cm3

694.3±65.0 °C at 760 mmHg

373.7±34.3 °C

0.0±2.3 mmHg at 25°C

1.609

0.27

93.07

1

9

5

34

659

2

O1C([H])([H])C([H])([H])N(C2C3C([H])=C([H])C(C4C([H])=C([H])C(=C(C([H])([H])O[H])C=4[H])OC([H])([H])[H])=NC=3N=C(N=2)N2C([H])([H])C([H])([H])OC([H])([H])[C@]2([H])C([H])([H])[H])[C@@]([H])(C([H])([H])[H])C1([H])[H]

KVLFRAWTRWDEDF-IRXDYDNUSA-N

InChI=1S/C25H31N5O4/c1-16-14-33-10-8-29(16)24-20-5-6-21(18-4-7-22(32-3)19(12-18)13-31)26-23(20)27-25(28-24)30-9-11-34-15-17(30)2/h4-7,12,16-17,31H,8-11,13-15H2,1-3H3/t16-,17-/m0/s1

[5-[2,4-bis[(3S)-3-methylmorpholin-4-yl]pyrido[2,3-d]pyrimidin-7-yl]-2-methoxyphenyl]methanol

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)

DMSO: ~50 mg/mL (~107.4 mM)
Water: <1 mg/mL
Ethanol: ~3 mg/mL (~6.4 mM)

配方 1 中的溶解度: ≥ 5 mg/mL (10.74 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 2 中的溶解度: ≥ 2.5 mg/mL (5.37 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 (5.37 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 (5.37 mM) (饱和度未知) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*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: ≥ 2.5 mg/mL (5.37 mM) (饱和度未知) in 5% DMSO + 95% (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。
*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 6 中的溶解度: 4% DMSO+30% PEG 300+ddH2O: 5mg/mL

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配方 7 中的溶解度: 50 mg/mL (107.40 mM) in 30 % SBE-β-CD (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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。