title: Novel chlorambucil-conjugated anionic linear-globular PEG-based second-generation dendrimer: in vitro/in vivo improved anticancer activity.
abstract: Evaluating the efficacy of anticancer drugs is an evolving and research-oriented issue. The objective of this study was to reduce the insolubility of chlorambucil (CBL) in water and improve the anticancer activity of CBL in vitro and in vivo through the conjugation of CBL with anionic linear-globular dendrimer (second generation, G2). In the current study, the anticancer activity among three groups that include CBL, CBL-G2 dendrimer, and control was measured in vitro and in vivo. In vitro studies showed that G2 anionic linear-globular polyethylene-glycol-based dendrimer, which conjugated to the CBL exterior through an ester linkage, was able to significantly improve the treatment efficacy over clinical CBL alone with respect to proliferation assay, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide; half maximal inhibitory concentration (IC50) was calculated to be 141 µg/mL for CBL alone and 27.7 µg/mL for CBL-G2 dendrimer; P<0.05. In addition, CBL-G2 dendrimer conjugate forestalled the growth of MCF-7 cancerous cells in addition to enhancing the number of apoptotic and necrotic cells as demonstrated by an annexin V-fluorescein isothiocyanate assay. CBL-G2 dendrimer conjugate was able to checkmate antiapoptotic Bcl-2 expression and Bcl-2/Bax ratio in a large scale compared with the control group and CBL alone (P<0.005). In vivo studies showed that tumor treatment by CBL-G2 dendrimer conjugate outstrips the efficacy of treatment compared with CBL alone. The evaluation was based on reduction in tumor volume and tumor growth inhibition of murine 4T1 mammary tumor cells. Tumor volume of 140%±8% was measured in the treatment with CBL-G2 dendrimer, whereas 152%±13.5% was calculated in the treatment with free CBL (P<0.05). However, there were no significant differences in histological assay among the three groups. In conclusion, tumor growth suppression potential of CBL-G2 dendrimer, which was assessed in both in vitro and in vivo experiments, has provided empirical evidence to buttress the fact that this compound could be considered for functional cancer treatment with low side effects.
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Artin
authors.initials: A
authors.lastname: Assadi
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Vahideh Sharifi
authors.initials: VS
authors.lastname: Najafabadi
authors.affiliation: Young Researchers and Elite Club, East Tehran Branch, Islamic Azad University.
authors.forename: Seyed Ataollah Sadat
authors.initials: SA
authors.lastname: Shandiz
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Azadah Shayeq
authors.initials: AS
authors.lastname: Boroujeni
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Sepehr
authors.initials: S
authors.lastname: Ashrafi
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Ali Zaman
authors.initials: AZ
authors.lastname: Vaziri
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Seyedeh Masoumeh
authors.initials: SM
authors.lastname: Ghoreishi
authors.affiliation: Department of Hepatitis and AIDS, Pasteur Institute of Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Aghasadeghi
authors.affiliation: Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Seyed Esmaeil Sadat
authors.initials: SE
authors.lastname: Ebrahimi
authors.affiliation: Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Morteza
authors.initials: M
authors.lastname: Pirali-Hamedani
authors.affiliation: Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences.
authors.forename: Mehdi Shafiee
authors.initials: MS
authors.lastname: Ardestani
keywords: CBL; G2; anionic linear-globular dendrimer; chlorambucil; in vitro cytotoxicity; in vivo efficacy