p110α (IC50 = 0.5 μM); p110δ (IC50 = 0.57 μM); p110β (IC50 = 0.97 μM); human CK2 (IC50 = 98 nM); human CK2α2 (IC50 = 3.869 μM); DNA-PK (IC50 = 1.4 μM)

LY294002 不仅对 PI3K 具有选择性；它也可能作用于其他脂质激酶和其他看似无关的蛋白质。除了 mTOR 和 DNA-PK 之外，LY294002 已被证明可以抑制 CK2（酪蛋白激酶 2）和 Pim-1 以及其他蛋白激酶。 LY294002 使 Akt/PKB 失活，从而触发细胞凋亡并抑制细胞增殖。在这些结肠癌细胞系中，LY294002 表现出显着的生长抑制和细胞凋亡诱导作用，同时磷酸化 Akt (Ser473) 的表达降低。 [2] 在肿瘤细胞中，LY294002 显着增加核固缩并减少细胞质体积。因此，LY294002 显着降低培养物中卵巢癌细胞的增殖。 LY294002 诱导细胞生长特异性 G1 期停滞，从而几乎完全抑制黑色素瘤细胞增殖，并部分抑制 MG-63（骨肉瘤细胞系）增殖。 LY294002 对细胞周期进程的影响可能揭示 PI3K 激活途径与癌细胞周期控制之间的潜在联系。 [3]

LY294002 在小鼠癌性腹膜炎模型中表现出显着的有效性，因为它还诱导细胞凋亡并抑制肿瘤生长，特别是在 LoVo 肿瘤中。 [2] LY294002 显着减少卵巢癌生长和腹水形成。[3] LY294002 显着减缓腹水和卵巢癌的发展。[3]

LY294002 对 PI3K 的抑制作用是使用纯化的重组酶和 1μM ATP 在放射测定中测定的。在室温 (24oC) 下，激酶反应持续一小时，然后通过添加 PBS 停止。然后，通过拟合可变斜率 S 形剂量反应曲线来计算 IC50 值。激酶选择性筛选用于确定 CK2 和 GSK3β（糖原合酶激酶 3β）的抑制作用。在 10μM ATP 中，根据 Upstate 激酶组对 LY294002 进行评估。

将1.0×105个细胞（100μL体积/孔）接种到96孔微量滴定板中。添加LY294002后，将一式三份的孔在37℃下培养0-48小时。处理后，将 10 μL Premix WST-1 添加到每个微培养孔中，然后将板在 37 °C 下孵育 60 分钟。此时，使用酶标仪在 450 nm 处进行吸光度测量。

Two groups of athymic nude mice (5–7 weeks) are inoculated i.p. with OVCAR-3 cells
0–100 mg/kg
Administered via i.p.

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Athymic mice were inoculated i.p. with the ovarian cancer cell line OVCAR-3. Seven days after inoculation, mice were treated with or without LY294002 (100 mg/kg of body weight) for 3 weeks. Body weight and abdominal circumference were measured twice weekly. At the end of the experiment, mice were sacrificed, ascites volume was measured, and tumors were excised. Mean tumor burden in the LY294002-treated group was reduced by approximately 65% versus controls. Virtually no ascites developed in the treatment group; mean volume of ascites in controls was 3.3 +/- 0.38 ml. OVCAR-3 cells also were cultured in vitro without and with LY294002 (1, 5, and 10 microM) for 24 h. The number of cells in 1, 5, and 10 microM LY294002-treated wells was reduced by 27, 56, and 75%, respectively, versus controls. In vivo and in vitro morphological studies demonstrated that LY294002 induced marked nuclear pyknosis and diminished cytoplasmic volume in the tumor cells, confirmed as apoptosis. Thus, LY294002 significantly inhibits growth and ascites formation of ovarian carcinoma in vivo and markedly inhibits ovarian cancer cell proliferation in vitro, suggesting an important role of PI3-K inhibitors as a potentially useful treatment for women with ovarian carcinoma.[3]

[1]. Biochem J. 2007 Jun 15;404(3):449-58.

[2]. Clin Cancer Res. 2002 Jun;8(6):1957-63.

[3]. Clin Cancer Res. 2000 Mar;6(3):880-6.

LY294002 is a chromone substituted with a phenyl group at position 8 and a morpholine group at position 2. It has a role as an EC 2.7.1.137 (phosphatidylinositol 3-kinase) inhibitor, an autophagy inhibitor and a geroprotector. It is a member of chromones, a member of morpholines and an organochlorine compound.
Specific inhibitor of phosphatidylinositol 3-kinase.
2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one has been reported in Hexagonia apiaria and Dietes bicolor with data available.
PI3K/BET Inhibitor LY294002 is a morpholine-based inhibitor of phosphatidylinositol 3-kinase (PI3K) and the bromodomain and extra-terminal (BET) family of proteins, with potential antineoplastic activity. Upon administration, the PI3K/BET inhibitor LY294002 specifically targets and binds to both PI3K and the acetylated lysine recognition motifs in the bromodomains of BET proteins. Inhibition of PI3K activity inhibits the PI3K/AKT kinase signaling pathway. This may result in inhibition of growth and survival for tumor cells in which the PI3K-mediated signaling pathway is overactivated. Inhibition of BET proteins prevents their interaction with acetylated histones, disrupts chromatin remodeling and inhibits the expression of oncogenic drivers that are important for cell proliferation and survival, which together may lead to an inhibition of proliferation in BET-overexpressing tumor cells. Activation of the PI3K signaling pathway is frequently associated with tumorigenesis. BET proteins, comprised of BRD2, BRD3, BRD4 and BRDT, are transcriptional regulators and play an important role during development and cellular growth. In tumor cells, BET proteins play a key role in the regulation of oncogene transcription and tumor cell proliferation.

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Purpose: Phosphatidylinositol 3'-kinase (PI3K) and Akt/protein kinase B(PKB) allow for escape from apoptosis in various human cancer cells. We postulated that 2-(4-morpholinyl)-8-phenyl-chromone (LY294002), a PI3K inhibitor, should inactivate Akt/PKB, consequently inhibiting cell proliferation and inducing apoptosis in vitro and in vivo. Experimental design: Human colon cancer cell lines (DLD-1, LoVo, HCT15, and Colo205) and their mouse xenografts (DLD-1 and LoVo) were used in this study. The expression of phosphorylated Akt (Ser(473)) and apoptosis in cancer cells were determined by immunoblotting and immunohistochemistry. To evaluate the activity of caspase-3 in culturing cells, the caspase colorimetric assay was also performed. Results: LY294002 demonstrated a remarkable growth-inhibitory and apoptosis-inducing effect in these colon cancer cell lines, with decreased expression of phosphorylated Akt (Ser(473)). However, there was a great discrepancy between the sensitivity for LY294002 and the level of expression of phosphorylated Akt. Although the LoVo and Colo205 cells exhibited high sensitivity to LY294002 with increased apoptosis, the DLD-1 and HCT15 cells did not show rapid induction of apoptosis. The caspase-3 activity was significantly high in the LoVo cells but not in the DLD-1 cells. In the experiments using mouse xenografts, we found that LY294002 administration in vivo also resulted in suppression of tumor growth and induction of apoptosis, especially in the LoVo tumors, and therefore showed remarkable effectiveness in the mouse peritonitis carcinomatosa model. Conclusions: PI3K-Akt/PKB plays an important role in colon cancer development and progression by helping to promote cell growth and allowing cells to escape apoptosis. These results propose the usefulness of LY294002 as an antitumoral agent for patients with colorectal cancer.[2]

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hosphatidylinositol 3-kinase (PI3-K) induces mitogenesis, cell growth, and cell transformation. Amplification of the gene encoding the P110alpha subunit likely is an important event in ovarian cancer progression, and PI3-K inhibitors are possible therapeutic agents for this disease. We evaluated effects of LY294002, a potent inhibitor of PI3-K, on growth of ovarian carcinoma in vivo and in vitro, and on ascites formation in vivo. Athymic mice were inoculated i.p. with the ovarian cancer cell line OVCAR-3. Seven days after inoculation, mice were treated with or without LY294002 (100 mg/kg of body weight) for 3 weeks. Body weight and abdominal circumference were measured twice weekly. At the end of the experiment, mice were sacrificed, ascites volume was measured, and tumors were excised. Mean tumor burden in the LY294002-treated group was reduced by approximately 65% versus controls. Virtually no ascites developed in the treatment group; mean volume of ascites in controls was 3.3 +/- 0.38 ml. OVCAR-3 cells also were cultured in vitro without and with LY294002 (1, 5, and 10 microM) for 24 h. The number of cells in 1, 5, and 10 microM LY294002-treated wells was reduced by 27, 56, and 75%, respectively, versus controls. In vivo and in vitro morphological studies demonstrated that LY294002 induced marked nuclear pyknosis and diminished cytoplasmic volume in the tumor cells, confirmed as apoptosis. Thus, LY294002 significantly inhibits growth and ascites formation of ovarian carcinoma in vivo and markedly inhibits ovarian cancer cell proliferation in vitro, suggesting an important role of PI3-K inhibitors as a potentially useful treatment for women with ovarian carcinoma.[3]

C19H17NO3

307.3432

307.12

C, 74.25; H, 5.58; N, 4.56; O, 15.62

154447-36-6

LY294002 hydrochloride;934389-88-5

3973

White to yellow solid powder

1.3±0.1 g/cm3

494.6±45.0 °C at 760 mmHg

182-184ºC

253.0±28.7 °C

0.0±1.3 mmHg at 25°C

1.627

3.82

42.68

0

4

2

23

463

0

O1CCN(C2=CC(=O)C3C=CC=C(C4C=CC=CC=4)C=3O2)CC1

CZQHHVNHHHRRDU-UHFFFAOYSA-N

InChI=1S/C19H17NO3/c21-17-13-18(20-9-11-22-12-10-20)23-19-15(7-4-8-16(17)19)14-5-2-1-3-6-14/h1-8,13H,9-12H2

2-morpholin-4-yl-8-phenylchromen-4-one

LY-294002; LY 294002; 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; LY-294002; 2-morpholino-8-phenyl-4H-chromen-4-one; 2-morpholin-4-yl-8-phenyl-4H-chromen-4-one; 2-morpholin-4-yl-8-phenylchromen-4-one; LY294002

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: ~36 mg/mL (117.1 mM)
Water: <1 mg/mL (slightly soluble or insoluble)
Ethanol: ~21 mg/mL (~68.3 mM)

配方 1 中的溶解度: 2.87 mg/mL (9.34 mM) 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), 悬浮液；超声助溶。
*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: ≥ 2.25 mg/mL (7.32 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 22.5 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 3 中的溶解度: ≥ 2.25 mg/mL (7.32 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 22.5mg/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 4 中的溶解度: ≥ 2.25 mg/mL (7.32 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 22.5 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。

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配方 5 中的溶解度: 4%DMSO+30%PEG 300+5%Tween 80+ddH2O: 5mg/mL

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配方 6 中的溶解度: 15.71 mg/mL (51.12 mM) in 0.5% CMC-Na/saline water (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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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。