(S)-3-(3,4-Dihydroxybenzyl) piperazine-2,5-dione (cyclo-Gly-L-DOPA or CG-Nio-CGLD) peptide loaded in Chitosan Glutamate-Coated Niosomes as anti-Colorectal cancer activity

Abstract

Background: Colorectal cancer (CRC), now the second most prevalent malignant tumor worldwide, is more prevalent in young adults. In recent decades, there has been progress in creating anti-colorectal cancer medications, including cytotoxic compounds.

Objectives: Novel anticancer drugs are needed to surmount existing obstacles. A recent study investigated the effectiveness of novel formulations in preventing colorectal cancer.

Methods: During this study, we assessed a new kind of niosome called cyclo-Gly-L-DOPA (CG-Nio-CGLD) made from chitosan glutamate. We evaluated the anti-colorectal cancer properties of CG-Nio-CGLD utilizing CCK-8, invasion assay, MTT assay, flow cytometry, and cell cycle analysis. The transcription of genes associated with apoptosis was analyzed using quantitative real-time PCR. At the same time, the cytotoxicity of nanomaterials on both cancer and normal cell lines was assessed using MTT assays. Novel anticancer drugs are needed to surmount existing obstacles. A recent study investigated the effectiveness of newly developed formulations in preventing colorectal cancer.

Results: The Nio-CGLD and CG-Nio-CGLD were spherical mean diameters of 169.12 ± 1.87 and 179.26 ± 2.17 nm, respectively. Entrapment efficiency (EE%) measurements of the Nio-CGLD and CG-Nio-CGLD were 63.12 ± 0.51 and 76.43 ± 0.34%, respectively. In the CG-Nio-CGLD group, the percentages of early, late, necrotic, and viable CL40 cells were 341.93%, 23.27%, 9.32%, and 25.48%. The transcription of the genes PP53, cas3, and cas8 was noticeably higher in the treatment group compared to the control group (P > 0.001). Additionally, the treatment group had lower BCL2 and survivin gene expression levels than the control group (P < 0.01). Additionally, CG-Nio-CGLD formulations demonstrated a biocompatible nanoscale delivery mechanism and displayed little cytotoxicity toward the CCD 841 CoN reference cell line.

Conclusion: These findings indicate that chitosan-based noisome encapsulation may enhance the effectiveness of CG-Nio-CGLD formulations in fighting cancer.

Keywords: Antiapoptotic activity; Cyclo-Gly-L-DOPA; Cytotoxicity; Niosome nanoparticles.

Fig. 1

A) The blending of Boc-Gly and L-DOPA-OMe hydrochloride, followed by the creation of cyclo-Gly-L-DOPA are the steps in the process. B) The chart of GC-MS analysis of cyclo-Gly-L-DOPA

Fig. 2

TEM, FeSEM electron microscopy images (A) and the FTIR spectra (B) of synthetic versions of Free noisome, CG-Nio, Nio-CGLD, and CG-Nio-CGLD. The images show the bilayer structure produced by the formulation of glutamate nanochitosan by Niosome

Fig. 3

(A) Drug-Release examination; (B) impact of storage time and temperature on the median diameter of Nio-CGLD, and CG-Nio-CGLD. Data represent the average of three separate studies, plus or minus one standard deviation. **p < 0.01, *p < 0.05. n = 3

Fig. 4

Malignant cell growth and invasion are inhibited in vitro by Nio-CGLD and CG-Nio-CGLD. In the CCK-8 test, it was shown that (A) free niosome, (B) CG-Nio, didn’t promoted cell proliferation but (C) Nio-CGLD and, (D) CG-Nio-CGLD promoted cell proliferation in CL40 and SW1417 cells

Fig. 5

Free noisome, CG-Nio, Nio-CGLD and CG-Nio-CGLD dilute solutions on SW1417 (A), CL40 (B), and CCD841 CoN (C) cell lines were examined for their effects on cell viability. (D) Flow cytometric investigation diagrams were shown in SW1417 and CL40 colorectal cancer cell lines. The cell cycle was regulated in SW1417 (E) and CL40 (F) colorectal cancer cell lines. Results are presented as mean ± SEM. The means with various superscript letters differ significantly (P ≤ 0.05)

Fig. 6

The effect of Nano-drug on cancer genes in CL40 and SW1417 cell line leads to induction of expression of pro-apoptotic genes P53, cas3, and cas8 at a significant level ***P < 0.001 and reduction of expression of anti-apoptotic genes SURVIVIN and BCL2 at a substantial level **P < 0.01. The GAPDH reference gene normalized data. There was no significant difference in the expression of pro/anti-apoptotic genes in the Blank control groups