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.
Cyclophosphamide monohydrate 是一种合成的DNA烷化剂，是DNA合成的抑制剂[1]。Cyclophosphamide monohydrate还能够可逆地抑制乙酰胆碱酯酶（AChE），其 IC50为511μM[2]。Cyclophosphamide monohydrate 在被细胞色素P-450酶代谢活化后，通过抑制DNA合成发挥抗肿瘤和免疫抑制作用，通常用于癌症和免疫相关研究[3][4]。
在体外实验中，用 Cyclophosphamide monohydrate（10–250μg/mL）处理单核巨噬细胞Raw 264.7降低了细胞活性，细胞毒性为69.58%，IC50为145.44μg/mL[5]。用Cyclophosphamide monohydrate（0.25–1mM；24h）处理人乳腺癌细胞MDA-MB-231和MDA-MB-435S的结果显示，Cyclophosphamide monohydrate通过诱导细胞表面CXCR4表达（不影响总CXCR4水平）和增加MMP9和MMP13的蛋白表达，增强了MDA-MB-231细胞的迁移能力，而对CXCR4表达较低的MDA-MB-435S细胞则无影响[6]。
在体内实验中，对携带小鼠结直肠肿瘤CT26的BALB/c小鼠进行Cyclophosphamide monohydrate（50mg/kg；一次；腹腔注射）和AdIL-12的联合治疗，诱导了超过50%的小鼠出现完全的肿瘤消退，显著延长了生存时间，减少了外周和脾脏中的CD4⁺CD25⁺Foxp3⁺ 调节性T细胞数量，增加了活化的树突状细胞数量，诱导了分泌IFN-γ的CD4 阳性T淋巴细胞，产生了强大的肿瘤特异性细胞毒性T淋巴细胞（CTL）反应，并降低了IL-10水平[7]。对携带小鼠乳腺癌肿瘤4T1的Balb/c小鼠口服给药Cyclophosphamide monohydrate（140mg/kg；每六天一次，持续十八天），显著抑制了肿瘤生长[8]。