Cyclophosphamide

Please contact us at for specific academic pricing.

Background

Cyclophosphamide, an inactive prodrug, is a kind of nitrogen mustard alkylating agent. Cyclophosphamide requires enzymatic and chemical activation. As a result, nitrogen mustard is produced. It causes DNA cross-linking that accounts for its cytotoxic properties. [1] IC50 of cytotoxicity in mouse embryo BALB/c 3T3 cells is 37.6 μM, [2] IC50 of cytotoxicity against human HL60 cells is 8.79 μM measured by MTT assay. [3]
Cyclophosphamide attaches the alkyl group to the guanine base of DNA causing its crosslinking, strand breakage and inducing mutations.
In vitro, cyclophosphamide has a dose-dependent, bimodal effect on the immune system. Low-dose cyclophosphamide not only decreases cell number but leads to decreased functionality of regulatory T cells (TREGs). Cyclophosphamide treatment enhances apoptosis and decreases homeostatic proliferation of these cells. Expression of GITR and FoxP3, which are involved in the suppressive activity of TREGs, is down-regulated after cyclophosphamide administration.[4] In primary human hepatocyte cultures, cyclophosphamide increases CYP3A4, CYP2C8, and CYP2C9 protein levels, causing its 4-hydroxylation rate enhance.[5] In somatic cells, cyclophosphamide produces gene mutations, chromosome aberrations, micronuclei and sister chromatid exchanges in a variety of cultured cells in the presence of metabolic activation as well as sister chromatid exchanges without metabolic activation. [6]
In vivo, it has produced chromosome damage and micronuclei in rats, mice and Chinese hamsters, and gene mutations in the mouse spot test and in the transgenic lacZ construct of Muta™Mouse. [6]

[1] Emadi A, Jones RJ, Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nat Rev Clin Oncol. 2009 Nov; 6 (11):638-47.
[2] Moon KY, Kwon CH. N3-methyl-mafosfamide as a chemically stable, alternative prodrug of mafosfamide. Bioorg Med Chem Lett. 1998 Jul 7; 8 (13):1673-8.
[3] Patel MM, Mali MD, Patel SK. Bernthsen synthesis, antimicrobial activities and cytotoxicity of acridine derivatives. Bioorg Med Chem Lett. 2010 Nov 1; 20 (21):6324-6.
[4] Lutsiak ME, Semnani RT, De Pascalis R,et al. Inhibition of CD4(+)25+ T regulatory cell function implicated in enhanced immune response by low-dose cyclophosphamide. Blood. 2005 Apr 1; 105 (7):2862-8. Epub 2004 Dec 9.
[5] Chang TK, Yu L, Maurel P, Waxman DJ. Enhanced cyclophosphamide and ifosfamide activation in primary human hepatocyte cultures: response to cytochrome P-450 inducers and autoinduction by oxazaphosphorines. Cancer Res. 1997 May 15; 57 (10):1946-54.
[6] Anderson D, Bishop JB, Garner RC, et al. Cyclophosphamide: review of its mutagenicity for an assessment of potential germ cell risks. Mutat Res. 1995 Aug; 330 (1-2):115-81.