| 生物活性 |
DHW-221 is a potent orally active dual PI3K/mTOR inhibitor, exhibiting low nanomolar potency against all four Class I PI3K isoforms and mTOR (PI3Kα, IC50 = 0.50 nM; PI3Kβ, IC50 = 1.9 nM; PI3Kγ, IC50 = 1.8 nM; PI3Kδ, IC50 = 0.74 nM; mTOR, IC50 = 3.9 nM). DHW-221 exerts antitumor effects by blocking the PI3K/Akt/mTOR pathway and inducing mitochondrial apoptosis and paraptosis (via Endoplasmic Reticulum (ER) stress and MAPK signaling) and arrests cell cycle, thereby inhibiting cell migration, invasion and angiogenesis. DHW-221 inhibits tumor growth in both the A549/Taxol (HY-B0015) and the HCC827 xenograft mouse models. DHW-221 can be used for non-small cell lung cancer (NSCLC), colon and breast cancer research[1][2][3].
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| IC50 & Target[1] |
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PI3Kα
0.50 nM (IC50)
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PI3Kβ
1.9 nM (IC50)
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PI3Kδ
0.74 nM (IC50)
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PI3Kγ
1.8 nM (IC50)
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mTOR
3.9 nM (IC50)
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体外研究
(In Vitro) |
DHW-221 (compound 8i) (72 小时) 对典型人类癌细胞表现出抗增殖活性，IC50 值分别为 0.36 μM (T47D)、0.14 μM (HCT116) 和 0.31 μM (MCF-7)[1]。
DHW-221 (0.3-3 μM) 以剂量依赖性方式降低 HCT116 细胞中的磷酸化 Akt (S473)，但对总蛋白 Akt 的表达无显著影响[1]。
DHW-221 (0-5 μM，2 周) 以剂量依赖性方式抑制 HCT116 细胞的增殖[1]。
DHW-221 (0.3-3 μM，0-48 小时) 以剂量依赖性方式抑制 HCT116 细胞迁移和侵袭[1]。
DHW-221 (0.3-3 μM) 以浓度依赖性方式诱导 HCT116 细胞凋亡，并伴有明显的核碎裂和染色质凝聚[1]。
DHW-221 能够与 PI3K 激酶的结合位点结合，与 Val882、Lys833 和 Thr887 位点形成氢键，与 Omipalisib (HY-10297) 类似，并与多个氨基酸残基形成 Pi-Pi 相互作用[1]。
DHW-221 (72 小时) 在 A549/Taxol (IC50 = 0.5274 μM) 和 A549 细胞 (IC50 = 0.4242 μM) 中表现出显著的浓度和时间依赖性细胞毒性[2]。
DHW-221 (0-2.4 μM，48-72 小时) 对 MDR 癌细胞 (A549 和 A549/Taxol 细胞) 具有显著的抑制活性，与对照组相比，48 小时后两种细胞均出现严重的细胞损伤[2]。
DHW-221 (0.15-2.4 μM，48 小时) 以浓度依赖性方式增加 A549/Taxol 细胞内 Rho-123 的积累，并显著下调 P-gp 的表达，表明其抑制 P-gp 的功能和蛋白表达[2]。
DHW-221 (50 nM，48 小时) 显著增强了 Taxol 的细胞毒作用，其作用类似于 P-gp 抑制剂 Verapamil (HY-14275)，表明其既可作为 P-gp 抑制剂，又可作为 MDR 逆转剂[2]。
DHW-221 (0.15-2.4 μM, 12-48 小时) 通过线粒体途径、ER 应激和 MAPK 信号通路引发 A549/Taxol 细胞凋亡和副凋亡[2]。
DHW-221 (0.15-2.4 μM, 48 小时) 通过下调细胞周期蛋白 cyclin D1、CDK4 和 CDK6 的表达，并上调 p21 的表达，将 A549/Taxol 和 A549 细胞的细胞周期阻滞于 G0/G1 期[2]。
DHW-221 (0.05-0.15 μM, 48 小时) 通过逆转上皮-间质转化 (EMT) 表型变化，抑制 A549/Taxol 细胞的迁移和侵袭能力[2]。
DHW-221 (0.15-2.4 μM，48 小时) 显著阻断 A549 和 A549/Taxol 细胞中的 PI3K/Akt 信号通路，表现为 PI3Kp110α 和磷酸化 Akt(p-Akt) 水平降低[2]。
DHW-221 (25-400 nM, 1-3 小时) 通过抑制 PI3K/HIF-1α/VEGF 信号轴来抑制 HUVEC 中的内皮管形成[3]。
DHW-221 (1.56-6.25 µM) 在体外大鼠主动脉环实验中抑制微血管出芽[3]。
DHW-221 (1.25-5 µM) 可抑制鸡胚绒毛尿囊膜中的新血管形成，且不影响胚胎活力[3]。
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
Cell Proliferation Assay[1]
| Cell Line: |
HCT116 cells |
| Concentration: |
0.3, 1 and 3 μM |
| Incubation Time: |
2 weeks |
| Result: |
Inhibited the formation of HCT116 cell clones in a concentration-dependent manner.
Almost completely inhibited the production of cell clones at 3 μM.
|
Cell Migration Assay [1]
| Cell Line: |
HCT116 cells |
| Concentration: |
0.3, 1 and 3 μM |
| Incubation Time: |
0, 24 and 48 h |
| Result: |
Its inhibition ability increased in a concentration-dependent manner.
|
Cell Invasion Assay[1]
| Cell Line: |
HCT116 cells |
| Concentration: |
0.3, 1 and 3 μM |
| Incubation Time: |
48 h |
| Result: |
Significantly inhibited HCT116 cell metastasis and invasiveness in a dose-dependent manner. |
Cell Viability Assay[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.075, 0.15, 0.3, 0.5, 1.2 and 2.4 μM |
| Incubation Time: |
24, 48 and 72 h |
| Result: |
Exhibited significant cytotoxicity in a concentration- and time-dependent manner in A549/Taxol and A549 cells with an resistance index (RI) of 1.2.
|
Cell Proliferation Assay[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
2 weeks |
| Result: |
Decreased the colony formation ability of A549/Taxol and A549 cells in a concentration-dependent manner.
|
Cell Cytotoxicity Assay[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
48 h |
| Result: |
Increased LDH release rates in A549 and A549/Taxol cells in a concentration-dependent manner.
|
Western Blot Analysis[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
3, 6, 12, 24 and 48 h |
| Result: |
Upregulated the expression levels of apoptosis-related proteins (cytochrome C, cleaved caspase 3, and cleaved PARP) in A549/Taxol cells in a concentration-dependent manner.
Downregulated the expression levels of the anti-apoptotic protein, Bcl-2, in A549/Taxol cells in a concentration-dependent manner.
Significantly inhibited the expression of Alix in A549/Taxol cells.
Significantly upregulated the expression levels of ATF4, CHOP (key regulator in ER stress), p-JNK, p-ERK, and p-p38 in A549/Taxol cells.
Decreased the expression levels of cyclin D1, CDK4, and CDK6 and increased p21 levels in A549/Taxol cells in a concentration-dependent manner.
Significantly decreased the expression levels of PI3Kp110a and p-Akt in a concentration-dependent manner, with no obvious change in the total Akt level in both cells.
Blocked the PI3K/Akt signaling pathway in both A549/Taxol and A549 cells.
Significantly downregulated p-FOXO3a (Ser253) expression and upregulated FOXO3a expression, accompanied by an increase in Bim expression in A549/Taxol cells.
Showed no significant changes in FOXO3a and Bim expression levels were observed in A549 cells.
Promoted FOXO3a accumulation in A549/Taxol cells when combined with MG132 (HY-13259).
Interfered with FOXO3a degradation in a proteasome-independent manner.
|
Immunofluorescence[2]
| Cell Line: |
A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
24 and 48 h |
| Result: |
Increased nuclear FOXO3a protein and decreased cytoplasmic phosphorylated FOXO3a at the Ser253 site.
Reversed the EMT phenotype, evidenced by enhanced fluorescence intensity of the epithelial marker occludin and weakened fluorescence of the mesenchymal marker vimentin.
|
Apoptosis Analysis[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
12 h (for vacuolation), and 48 h (for apoptosis/MMP) |
| Result: |
Significantly increased the rates of early and late apoptosis in both A549/Taxol and A549 cells after 48 h.
Its pro-apoptotic ability in A549/Taxol cell was stronger than that of A549 cells at 2.4 μM.
Triggered apoptosis in A549/Taxol and A549 cells in a concentration-dependent manner.
Decreased the intracellular MMP in A549/Taxol cells after 48 h.
Induced the formation of massive vacuoles in A549/Taxol cells at 2.4 μM after 12 h.
Attenuated cytoplasmic vacuolation when combined with Cycloheximide (HY-12320, 20 μM).
|
Cell Cycle Analysis[2]
| Cell Line: |
A549/Taxol cells |
| Concentration: |
0.15, 0.60 and 2.40 μM |
| Incubation Time: |
48 h |
| Result: |
Significantly reduced the proportion of cells in S phase and increased the proportion of cells in G0/G1 phase. |
Cell Migration Assay [2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.05, 0.10 and 0.15 μM |
| Incubation Time: |
0, 24 and 48 h |
| Result: |
Significantly decreased the wound healing rate compared to the control group in both A549 and A549/Taxol cells. |
Cell Invasion Assay[2]
| Cell Line: |
A549 and A549/Taxol cells |
| Concentration: |
0.05, 0.10 and 0.15 μM |
| Incubation Time: |
24 h |
| Result: |
Significantly reduced the number of cells that migrated and invaded through the chamber in A549/Taxol cells. |
Western Blot Analysis[2]
| Cell Line: |
A549/Taxol cells |
| Concentration: |
0.05, 0.10 and 0.15 μM |
| Incubation Time: |
48 h |
| Result: |
Increased the expression levels of E-cadherin and occludin but significantly decreased the expression levels of mesenchymal markers, including vimentin and slug.
Decreased the expression of transcription factor snail.
Significantly downregulated the expression levels of MMP2 and MMP9 in A549/Taxol cells.
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药代动力学
(Parmacokinetics)[3] |
| Species |
Dose |
Route |
Indicator |
value |
| Mice |
10 mg/kg |
i.v. |
T1/2 |
3.7 h |
| Mice |
10 mg/kg |
p.o. |
T1/2 |
7.1 h |
| Mice |
10 mg/kg |
i.v. |
Tmax |
0.083 h |
| Mice |
10 mg/kg |
p.o. |
Tmax |
3.0 h |
| Mice |
10 mg/kg |
i.v. |
Cmax |
67395.0 ng/mL |
| Mice |
10 mg/kg |
p.o. |
Cmax |
11761.7 ng/mL |
| Mice |
10 mg/kg |
i.v. |
AUCall |
261501.9 ng·h/mL |
| Mice |
10 mg/kg |
p.o. |
AUCall |
176495.5 ng·h/mL |
| Mice |
10 mg/kg |
i.v. |
MRTINF_obs |
4.7 h |
| Mice |
10 mg/kg |
p.o. |
MRTINF_obs |
11.4 h |
| Mice |
10 mg/kg |
p.o. |
F |
67.5 % |
|
体内研究
(In Vivo) |
DHW-221 (10、20 和 40 mg/kg，灌胃，每日一次，持续 2 周) 通过 FOXO3a 核转位，在 A549/Taxol 荷瘤小鼠模型中抑制肿瘤生长，且未引起体重变化和毒性[2]。
DHW-221 (10、20 和 40 mg/kg，口服，每日一次，共 21 天) 可抑制 HCC827 异种移植小鼠模型中的肿瘤生长[3]。
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
| Animal Model: |
Male nude mice (4-6 weeks old) intravenously injected with A549/Taxol cells[2] |
| Dosage: |
25 and 50 mg/kg |
| Administration: |
i.g., q.d. for 2 weeks |
| Result: |
Significantly decreased the number of lung nodules.
Downregulated p-FOXO3a expression and upregulated FOXO3a expression.
Showed no change in Ki67 expression.
Showed no significant differences in the body weight and viscera index compared to the control group.
Caused no behavioral abnormalities or loss of appetite.
Exhibited no evident histological abnormalities in the heart, liver, kidney, or spleen.
Resulted in no abnormal toxicity signs in serum biochemical parameters (CK, ALT, AST, CRE), indicating no cardiac, hepatic, or renal toxicity.
|
| Animal Model: |
Nude mice bearing HCC827 xenografts (Subcutaneous and orthotopic injection)[3] |
| Dosage: |
10, 20 and 40 mg/kg |
| Administration: |
p.o., q.d. for 21 days |
| Result: |
Significantly inhibited tumor growth in both subcutaneous and orthotopic models.
Showed no significant effect on body weight.
Exhibited only mild liver toxicity at the high dose (40 mg/kg).
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| 分子量 |
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| Formula |
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| CAS 号 |
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| 运输条件 |
Room temperature in continental US; may vary elsewhere.
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| 储存方式 |
Please store the product under the recommended conditions in the Certificate of Analysis.
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| 纯度 & 产品资料 |
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| 参考文献 |
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