Colon targeted dosage form of Capecitabine using folic acid anchored modified carbon nanotube: in vitro cytotoxicity, apoptosis and in vivo roentgenographic study

Abstract

Objective

Development of dosage form comprising of Capecitabine loaded carbon nanotubes for its targeted delivery to the colon.

Method

Single walled carbon nanotubes (SWCNT) were functionalized by -COOH and Chitosan along with Folic acid. Capecitabine was loaded in these SWCNT’s, and the system was analyzed by FTIR, SEM and Raman spectroscopy. Percent drug loading was assessed and the cytotoxicity (COLO320DM and HT29) was verified by using MTT and SRB assay. The apoptosis study was carried out by flowcytometry. The system was enclosed in an enteric coated capsule with pH sensitive polymers and characterized for invitro disintegration, dissolution and invivo roentgenographic studies.

Results

FTIR, Raman and XRD studies indicated the confirmation of attachments, whereas SEM exhibited size range of 200-500 nm. Drug loading capacity was observed to be 94.63 ± 1.07%. Cytotoxicity studies of Capecitabine and FA–CHI-F-SWCNT-Capecitabine against COLO320DM by using MTT assay showed that FA–CHI-F-SWCNT- Capecitabine exhibited 86.45 ± 0.5788% inhibition whereas pure Capecitabine showed 50.52 ± 0.3106% inhibition. Against HT29, the % inhibition was observed to be 82.76 ± 0.4668% and 56.41 ± 0.2316% respectively for FA–CHI-F-SWCNT-Capecitabine and pure Capecitabine. In case of SRB assay of COLO320DM, the FA–CHI-F-SWCNT-Capecitabine exhibited 89.62 ± 0.4095% inhibition and Capecitabine showed 84.36 ± 0.2559% inhibition, whereas against HT29, FA–CHI-F-SWCNT-Capecitabine showed 81.36 ± 0.2958% inhibition and Capecitabine exhibited 90.62 ± 0.4196% inhibition.

Conclusion

FA–CHI-F-SWCNT loaded system revealed better cytotoxicity as compared with pure Capecitabine against two different cell lines. Invivo studies revealed that the prepared capsule formulation remained intact in the stomach thereby preventing drug release in the gastric milieu.

Graphical Abstract

Highlights

• Folic acid anchored single walled carbon nanotube to target specific folate receptors which are over expressed in colon cancer.

• In vitro cytotoxicity against colon cancer cell lines COLO320DMm, HT29 gave better results as compared to pure capacitabine give guideline for in vivo study.

• Apoptosis study by flow cytometry showed early death of cancer cell as compared with control

• In vivo roentgenographic study on rabbit was carried out successfully which further open arena of pharmacokinetic biodistribution study.

• Colon targeted enteric coated capsule filled with Carbon nanotube loaded capacitabine with folic acid gives exclusive release in the colonic region without premature release in stomach. Provide platform for targeting any organ tissue specifically.

• This study provides idea for use of carbon nanotube in biologicals with minimizing side effects of carrier CNT as well chemotherapeutics in cancer therapy. Also provide data for study on volunteer.

Keywords:

• Carbon nanotube
• Capecitabine
• colorectal cancer
• cytotoxicity
• roentgenography
• Flowcytometry

Acknowledgements

We highly acknowledge Department of BCUD, Shivaji University Kolhapur, for providing grant to carry out this work under Research Initiation Scheme, 2017. The authors are also thankful to the Hon. Secretary KE society Kasegaon, and the Principal Rajarambapu College of Pharmacy, Kasegaon, Sangli, Maharashtra, for providing facilities. We also acknowledge the help of Bharti Vidyapeeth College of Pharmacy, Kolhapur for providing necessary research facilities. We are also thankful to Biocyte Institute of Research and Development (BiRD) Sangli (MS) for their help and NCCS, Pune for providing the cell lines required in the studies.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work is funded by Shivaji University Kolhapur, Maharashtra, India under Research Initiation Shame 2017, for young researchers Department of BCUD SUK, No- SU/C&UD Section/83/239 date–10/05/1018.