Ifosfamide is an alkylating chemotherapeutic agent with activity against a wide range of tumors.

DNA Alkylator^[1]

Ifosfamide is an alkylating chemotherapeutic agent with activity against a wide range of tumors^[1]. Ifosfamide is activated by the cytochrome P450 family. Ifosfamide (0-5 mM) suppresses the viability of CYP2B1-expressing C8III-1 cells, while it is cytotoxic to the non-CYP2B1-expressing CrFK cells only at high concentration (5 mM)^[2]. CYP BM3 mutants activates Ifosfamide, and Ifosfamide shows inhibitory activity against human U2OS cells^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Ifosfamide (50 mg/kg, i.p.) treatment before mating increases percentage of post-implantation loss and resorbs fetuses in pregnant rats. Ifosfamide also (50 mg/kg, i.p.) decreases the progesterone in the serum of pregnant rats. However, Ifosfamide causes no obvious difference with the control rats at 25 mg/kg. Ifosfamide (25, 50 mg/kg, i.p.) induces apoptosis and histological changes in the placentas and prenatal rats, most sensitive fetal organs are the brain, liver and kidney^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

261.09

C₇H₁₅Cl₂N₂O₂P

3778-73-2

异环磷酰胺

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*该产品在溶液状态不稳定，建议您现用现配，即刻使用。

DMSO : ≥ 50 mg/mL (191.50 mM)

* “≥” means soluble, but saturation unknown.

请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液；该产品在溶液状态不稳定，建议您现用现配，即刻使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 0.5% CMC-Na/saline water

  Solubility: 25 mg/mL (95.75 mM); Clear solution; Need ultrasonic
• 2.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (9.58 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (9.58 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 3.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (9.58 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (9.58 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 4.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (9.58 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (9.58 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Helal M. Prenatal effects of transplacental exposure to ifosfamide in rats. Biotech Histochem. 2016 Jul;91(5):357-68.

  [2]. Karle P, et al. Necrotic, rather than apoptotic, cell death caused by cytochrome P450-activated ifosfamide. Cancer Gene Ther. 2001 Mar;8(3):220-30.

  [3]. Vredenburg G, et al. Activation of the anticancer drugs cyclophosphamide and ifosfamide by cytochrome P450 BM3 mutants. Toxicol Lett. 2015 Jan 5;232(1):182-92.

In a 3-cm dish, 4 × 10⁴ cells are seeded in 2 mL of medium. The next day, Ifosfamide is added to final concentrations from 0 to 5 mM. After six additional days the medium is removed, the cells washed with PBS, and either stained or counted^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Rats^[1]
Prior to mating, female rats are divided into four groups of eight: group 1, untreated negative controls; group 2, rats injected i.p. with 1 mL 0.9% NaCl; group 3, rats injected i.p. with 25 mg/kg Ifosfamide; group 4, rats injected i.p. with 50 mg/kg Ifosfamide. After injection of Ifosfamide once daily for 5 consecutive days, three females are placed in a cage with one untreated male for up to 1 week. Vaginal smears are examined daily to determine pregnancy. The first 24 h period following mating is designated day one of pregnancy if sperm are detected. The pregnant females are housed singly and observed daily for signs of toxicity and abortion. All pregnant animals are sacrificed by decapitation on day 18 of gestation. Cardiac blood (2.5 -3 mL/rat) is collected in nonheparinized test tubes, centrifuged at 3,000× g for 30 min and the serum is decanted and stored at -70°C until used for hormone assay. After blood collection, uteri and both ovaries are removed, washed in saline solution and the corpora lutea of pregnancy are counted visually, and the number of implantation sites, resorption sites and viable fetuses are counted in each uterine horn. Each fetus is removed from its umbilical cord, weighed and the crown rump (CR) length is measured. Fetuses are examined for external malformation and the placental weights are recorded. Fetuses and placentas of control and treated groups are fixed in 10% neutral buffered formalin for immunohistochemical and histological examination^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Helal M. Prenatal effects of transplacental exposure to ifosfamide in rats. Biotech Histochem. 2016 Jul;91(5):357-68.

  [2]. Karle P, et al. Necrotic, rather than apoptotic, cell death caused by cytochrome P450-activated ifosfamide. Cancer Gene Ther. 2001 Mar;8(3):220-30.

  [3]. Vredenburg G, et al. Activation of the anticancer drugs cyclophosphamide and ifosfamide by cytochrome P450 BM3 mutants. Toxicol Lett. 2015 Jan 5;232(1):182-92.