1. Cell Cycle/DNA Damage
  2. DNA Alkylator/Crosslinker
  3. Cyclophosphamide hydrate

Cyclophosphamide hydrate (Synonyms: Cyclophosphamide monohydrate)

Cat. No.: HY-17420A Purity: >98.0%
Handling Instructions

Cyclophosphamide hydrate is a synthetic alkylating agent chemically related to the nitrogen mustards with antineoplastic and immunosuppressive activities.

For research use only. We do not sell to patients.

Cyclophosphamide hydrate Chemical Structure

Cyclophosphamide hydrate Chemical Structure

CAS No. : 6055-19-2

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Based on 1 publication(s) in Google Scholar

Other Forms of Cyclophosphamide hydrate:

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Description

Cyclophosphamide hydrate is a synthetic alkylating agent chemically related to the nitrogen mustards with antineoplastic and immunosuppressive activities.

In Vitro

Cyclophosphamide induces outer membrane blebbing, leads to DNA fragmentation, as revealed by TUNEL staining of free 3'-OH DNA ends, and induces cleavage of the caspase 3 and caspase 7 substrate PARP in 9L/P450 cells. Bcl-2 expression fully blocks the activation of both initiator caspases as well as the effector caspase 3 in cells treated with activated Cyclophosphamide. Bcl-2 inhibits the cytotoxic effects but not the cytostatic effects of activated Cyclophosphamide[1]. Cyclophosphamide inhibits the AChE reversibly with an IC50 of 511 μM[2]. Carbon tetrachloride does not affect the direct cytotoxicity of cyclophosphamide or 4-hydroxycyclophosphamide to cells in culture[3].

Clinical Trial
Storage
Powder -20°C 3 years
  4°C 2 years

*The compound is unstable in solutions, freshly prepared is recommended.

Solvent & Solubility
In Vitro: 

H2O : ≥ 50 mg/mL (179.15 mM)

DMSO : ≥ 38 mg/mL (136.15 mM)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 3.5829 mL 17.9147 mL 35.8295 mL
5 mM 0.7166 mL 3.5829 mL 7.1659 mL
10 mM 0.3583 mL 1.7915 mL 3.5829 mL
*Please refer to the solubility information to select the appropriate solvent.
References
Kinase Assay
[1]

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.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[1]

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.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

279.10

Formula

C₇H₁₇Cl₂N₂O₃P

CAS No.

6055-19-2

SMILES

ClCCN(CCCl)P1(OCCCN1)=O.[H]O[H]

Shipping

Room temperature in continental US; may vary elsewhere

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Product Name:
Cyclophosphamide hydrate
Cat. No.:
HY-17420A
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Cyclophosphamide hydrate

Cat. No.: HY-17420A