| 名稱 | Cyclophosphamide hydrate |
| 描述 | Cyclophosphamide hydrate is a DNA alkylating agent, an inhibitor of DNA synthesis. Cyclophosphamide hydrate has antitumor and immunosuppressive activities. |
| 細(xì)胞實(shí)驗(yàn) | 9L/pBabe, 9L/Bax, and 9L/Bcl-2 cells are treated with 12, 24, or 50 μM MFA for 72 h. Cells remaining on the plates at 0, 24, 48, and 72 h are washed twice with cold PBS and then stained for 5 min with crystal violet [1.25 g of crystal violet dissolved in a solution containing 50 mL of 37% formaldehyde and 450 mL of methanol]. The stained cells are washed three times in tap water and the plates are allowed to dry. The stain is eluted from the cells with 70% ethanol and the absorbance is then read at 595 nm. The staining intensity of each drug-treated sample (A?595) is then graphed as a percentage of the staining intensity at the 0-h time point. |
| 激酶實(shí)驗(yàn) | 9L cells are treated with drug for the times indicated in each experiment. Floating and attached cells are collected, pooled, resuspended in lysis buffer (10 mM HEPES buffer, pH 7.4, containing 2 mM EDTA, 0.1% CHAPS detergent, 5 mM DTT, 350 ng/mL phenylmethylsulfonyl fluoride, 10 ng/mL pepstatin A, 10 ng/mL aprotinin, and 20 ng/mL leupeptin) and lysed by three freeze-thaw cycles (alternating between a dry ice isopropanol bath and a 37°C water bath). Lysates are spun in a bench top centrifuge at full speed for 15 min and the supernatant (cell extract) fraction transferred to a new tube. Cell extracts (20 μL) are assayed for caspase 9, caspase 8, and caspase 3 activity by incubation at 37°C for either 1 h (caspase 3) or 3 h (caspase 9 and caspase 8) in 500 μL of reaction buffer (10 mM HEPES, pH 7.4, 2 mM EDTA, 0.1% CHAPS, and 5 mM DTT) containing 50 μM caspase form-selective substrate: Ac-LETD-AFC for caspase 8; Ac-LEHD-AFC for caspase 9; and Ac-DEVD-AMC for caspase 3. Background activity is determined for each sample as follows. Cell extracts are preincubated for 15 min at room temperature, with or without caspase form-selective inhibitor: 1 μM z-LETD-FMK for caspase 8, 1 μM z-LEHD-FMK for caspase 9, and 5 μL of Casputin for caspase 3. Caspase activity measured in the absence of inhibitor is divided by the background caspase activity measured in the presence of inhibitor. A value of 1 is subtracted from each measured activity, such that a caspase activity of 0 corresponds to no increase in the specific caspase activity with drug treatment. Fluorescence of the caspase product (excitation at 395 nm and emission at 525 nm for AFC substrates, and excitation at 380 nm and emission at 460 nm for the AMC substrate) is measured using a Shimadzu model RF-1501 spectrofluorophotometer and the manufacturer's PC-1501 software package. |
| 體外活性 | 方法：巨噬細(xì)胞 Raw 264.7 用 Cyclophosphamide hydrate (10-250 μg/mL) 處理 48 h，使用 MTT assay 檢測(cè)細(xì)胞活力。
結(jié)果：Cyclophosphamide hydrate 對(duì) Raw 264.7 有細(xì)胞毒性，IC50 為 145.44 μg/mL。[1]
方法：人乳腺癌細(xì)胞 MDA-MB-231 和 MDA-MB-435S 用 Cyclophosphamide hydrate (0.25-1 mM) 處理 4-24 h，使用 Wound healing assay 檢測(cè)細(xì)胞遷移情況。
結(jié)果：遷移的 MDA-MB-231 細(xì)胞數(shù)量的增加取決于 Cyclophosphamide 的濃度，而到 MDA-MB-435S 細(xì)胞的遷移顯著減少，并且與 Cyclophosphamide 濃度無(wú)關(guān)。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Cyclophosphamide hydrate (50-150 mg/kg) 單次腹腔注射給攜帶小鼠結(jié)直腸腫瘤 CT26 的 BALB/c 小鼠。
結(jié)果：Cyclophosphamide hydrate 誘導(dǎo)腫瘤體積顯著減少。[3]
方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Cyclophosphamide hydrate (140 mg/kg) 口服給藥給攜帶小鼠乳腺癌腫瘤 4T1 的 Balb/c 小鼠，每六天一次，持續(xù)十八天。
結(jié)果：Cyclophosphamide hydrate 治療顯著抑制了小鼠的腫瘤生長(zhǎng)。[4] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | Shipping with blue ice. |
| 溶解度 | DMSO : 50 mg/mL (179.15 mM), The compound is unstable in solution. Please use soon.
H2O : 7 mg/mL (25.08 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | DNA Alkylator/Crosslinker | Cyclophosphamide Monohydrate | inhibit | Cyclophosphamide | Cyclophosphamide hydrate | Cyclophosphamide Hydrate | Inhibitor |
| 相關(guān)產(chǎn)品 | Crystal Violet | Benzocaine | Flurbiprofen | Cisplatin | N-Nitroso-N-methylurea | Clarithromycin | Methoxsalen | Temozolomide | Busulfan | Cholesterol | Prednisone | Carboplatin |