Korean J Physiol Pharmacol.  2023 Jan;27(1):31-38. 10.4196/kjpp.2023.27.1.31.

Characterization of a conjugated polysuccinimide-carboplatin compound

• Lee SY
• Chae CH
• Zrínyi M
• Che X
• Choi JY
• Cho DH

• KMID: 2537501
• DOI: http://doi.org/10.4196/kjpp.2023.27.1.31

Abstract

Carboplatin, an advanced anticancer drug with excellent efficacy against ovarian cancer, was developed to alleviate the side effects that often occur with cisplatin and other platinum-based compounds. Our study reports the in vitro characteristics, viability, and activity of cells expressing the inducible nitric oxide synthase (iNOS) gene after carboplatin was conjugated with polysuccinimide (PSI) and administered in combination with other widely used anticancer drugs. PSI, which has promising properties as a drug delivery material, could provide a platform for prolonging carboplatin release, regulating its dosage, and improving its side effects. The iNOS gene has been shown to play an important role in both cancer cell survival and inhibition. Herein, we synthesized a PSI-carboplatin conjugate to create a modified anticancer agent and confirmed its successful conjugation. To ensure its solubility in water, we further modified the structure of the PSI-carboplatin conjugate with 2-aminoethanol groups. To validate its biological characteristics, the ovarian cancer cell line SKOV-3 and normal ovarian Chinese hamster ovary cells were treated with the PSI-carboplatin conjugate alone and in combination with paclitaxel and topotecan, both of which are used in conventional chemotherapy. Notably, PSI-carboplatin conjugation can be used to predict changes in the genes involved in cancer growth and inhibition. In conclusion, combination treatment with the newly synthesized polymer-carboplatin conjugate and paclitaxel displayed anticancer activity against ovarian cancer cells but was not toxic to normal ovarian cancer cells, resulting in the development of an effective candidate anticancer drug without severe side effects.

Keyword

Carboplatin; Conjugation; iNOS; Ovarian cancer; Polysuccinimide

Figure

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  Fig. 1 Synthesis of polysuccinimide (PSI) and polyaspartic acid (PSAP). (A) Thermal polycondensation of aspartic acid into polysuccinimide. (B) Ring-opening reaction of polysuccinimide to form poly(aspartic acid).

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  Fig. 2 Synthesis of PSI-conjugated compounds. (A) Synthesis of the PSI-carboplatin conjugate with aminoethanol modifying groups. (B) Synthesis of the PSI-carboplatin conjugate with 4-dimethylaminopyridine (DMAP) modifying groups.

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  Fig. 3 1H NMR spectra of (A) carboplatin, (B) PSI-carboplatin, and (C) PSI-2-aminoethanol. PSI, polysuccinimide.

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  Fig. 4 Results of cell viability assays. (A) Viability of SKOV-3 cancer cells determined by MTS assay. (B) Viability of normal CHO cells determined by MTS assay. OD values are presented as the mean ± SE in each treatment group. “*” denotes a statistically significant difference (p < 0.05) vs. the control group. PSI, polysuccinimide; MTS, 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium; CHO, Chinese hamster ovary; OD, optical density.

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  Fig. 5 Results of iNOS mRNA and Western blot analyses. (A) iNOS mRNA expression in SKOV-3 cancer cells. iNOS mRNA levels are presented as median values with interquartile ranges. Relative expression was determined based on the ΔΔCt method. “*” denotes a statistically significant difference (p < 0.05) vs. the control group. (B) Western blotting results. (C) Densitometry analysis of the Western blotting results. Relative iNOS protein levels are expressed as the mean relative quantities (RQ values) ± SE based on comparisons between the control (n = 3) and treatment groups (n = 3). iNOS, inducible nitric oxide synthase; PSI, polysuccinimide.

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