Cyclophosphamide monohydrate

Cyclophosphamide monohydrate is a synthetic DNA alkylating agent, an inhibitor of DNA synthesis^[1]. Cyclophosphamide monohydrate also inhibited the AChE reversibly with an IC₅₀ of 511μM^[2]. Cyclophosphamide monohydrate, after being metabolized by cytochrome P-450 enzyme to become active, exerts anti-tumor and immunosuppressive effects by inhibiting DNA synthesis, and is usually used in cancer and immune-related researches ^[3][4].

In vitro, treatment of Monocyte Macrophage Cells Raw 264.7 with Cyclophosphamide monohydrate (10–250μg/mL) reduced cell viability, with a cell cytotoxicity of 69.58% and an IC₅₀ of 145.44μg/mL^[5]. Treatment of human breast cancer cells MDA-MB-231 and MDA-MB-435S with Cyclophosphamide monohydrate (0.25–1mM; 24h) revealed that Cyclophosphamide monohydrate enhanced the migration ability of MDA-MB-231 cells by inducing cell-surface CXCR4 expression (without affecting total CXCR4 levels) and increasing the protein expression of MMP9 and MMP13, while having no effect on MDA-MB-435S cells with low CXCR4 expression^[6].

In vivo, combination treatment of Cyclophosphamide monohydrate (50mg/kg; once; i.p.) and AdIL-12 in BALB/c mice carrying mouse colorectal tumor CT26 induced complete tumor regression in >50% of mice, significantly prolonged survival, reduced CD4⁺CD25⁺Foxp3⁺ regulatory T cells, increased activated dendritic cells, induced IFN-γ-secreting CD4-positive T lymphocytes, generated a powerful tumor-specific CTL response, and reduced IL-10 levels^[7]. Oral administration of Cyclophosphamide monohydrate (140mg/kg; once every six days for eighteen days) to Balb/c mice carrying mouse breast cancer tumor 4T1 significantly inhibited tumor growth^[8].

References:
[1] Schwartz PS, Waxman DJ. Cyclophosphamide induces caspase 9-dependent apoptosis in 9L tumor cells. Mol Pharmacol. 2001;60(6):1268-1279.
[2] al-Jafari AA, Duhaiman AS, Kamal MA. Inhibition of human acetylcholinesterase by cyclophosphamide. Toxicology. 1995;96(1):1-6.
[3] Liu P, Jaffar J, Hellstrom I, Hellstrom KE. Administration of cyclophosphamide changes the immune profile of tumor-bearing mice. J Immunother. 2010;33(1):53-59.
[4] Emadi A, Jones RJ, Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nat Rev Clin Oncol. 2009;6(11):638-647.
[5] Yadav A, Mandal MK, Dubey KK. In Vitro Cytotoxicity Study of Cyclophosphamide, Etoposide and Paclitaxel on Monocyte Macrophage Cell Line Raw 264.7. Indian J Microbiol. 2020;60(4):511-517.
[6] Hung CM, Hsu YC, Chen TY, Chang CC, Lee MJ. Cyclophosphamide promotes breast cancer cell migration through CXCR4 and matrix metalloproteinases. Cell Biol Int. 2017;41(3):345-352.
[7] Malvicini M, Rizzo M, Alaniz L, et al. A novel synergistic combination of cyclophosphamide and gene transfer of interleukin-12 eradicates colorectal carcinoma in mice. Clin Cancer Res. 2009;15(23):7256-7265.
[8] Shi H, Hou B, Li H, Zhou H, Du B. Cyclophosphamide Induces the Ferroptosis of Tumor Cells Through Heme Oxygenase-1. Front Pharmacol. 2022;13:839464.