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Cliv-92-Loaded Glycyrrhetinic Acid-Modified Chitosan Nanoparticles for Enhanced Hepatoprotection–Preparation, Characterization, and In Vivo Evaluation

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Abstract

Cliv-92 is a mixture of three structurally similar coumarinolignoids and a proven hepatoprotective agent. Low aqueous solubility and poor bioavailability are notable hindrances for its further use. Therefore, glycyrrhetinic acid-linked chitosan nanoparticles loaded with Cliv-92 were prepared for active targeting to the liver. The nanoparticles were prepared by the ionic gelation method to avoid the use of toxic solvents/rigorous agitation. The method of preparation was optimized using a central composite design with independent variables, namely polymer: drug ratio (3:1, w/w), crosslinker concentration (0.5%), and stirring speed (750 rpm). The optimized nanoparticles had a mean particle size of 185.17 nm, a polydispersity index of 0.41, a zeta potential of 30.93 mV, and a drug loading of 16.30%. The prepared formulation showed sustained release of approximately 63% of loaded Cliv-92 over 72 h. The nanoparticles were freeze-dried for long-term storage and further characterized. The formulation was found to be biocompatible for parenteral delivery. In vivo imaging study showed that optimized nanoparticles were preferentially accumulated in the liver and successfully targeting the liver. The present study successfully demonstrated the improved pharmacokinetic properties (≈12% relative bioavailability) and efficacy profile (evidenced by in vivo and histopathological studies) of fabricated Cliv-92 nanoparticles.

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Abbreviations

ALB:

Albumin

ALP:

Alkaline phosphatase

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

BIL:

Total bilirubin

CAT:

Catalase

CDCl3 :

Chloroform-d

CH3COOD:

Acetic (acid-d)

CNP:

Cliv-92 nanoparticles

CPCSEA:

Committee for the Purpose of Control and Supervision of Experiments on Animals

CTS:

Chitosan

DPX:

Dibutylphthalate polystyrene xylene

DSC:

Differential scanning calorimetry

EDC·HCl:

N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride

FTIR:

Fourier transform infrared

GA:

Glycyrrhetinic acid

GA-CTS complex:

Glycyrrhetinic acid-modified chitosan

GPx:

Glutathione peroxidase

GR:

Glutathione reductase

GSH:

Reduced glutathione

GST:

Glutathione S-transferase

H&E:

Hematoxylin and eosin

ICG:

Indocyanine green

ICMR:

Indian Council of Medical Research

IL:

Interleukin

KBr:

Potassium bromide

LDH:

Lactate dehydrogenase

NaCl:

Sodium chloride

NaOH:

Sodium hydroxide

NDDS:

Novel drug delivery systems

NHS:

N-hydroxysuccinimide

NMR:

Nuclear magnetic resonance

PBS:

Phosphate-buffered saline

RT:

Room temperature

SOD:

Superoxide dismutase

TNF-α :

Tumor necrosis factor-α

TP:

Total proteins

TPP:

Sodium tripolyphosphate

UV–Vis:

Ultraviolet–visible

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Acknowledgements

KSY is thankful to the Indian Council of Medical Research, New Delhi, for awarding Senior Research Fellowship. The authors are grateful to the CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow for providing necessary facilities. The authors would like to acknowledge the support of the Malaviya National Institute of Technology, Jaipur for the TEM analysis, and the Indian Institute of Technology, Kanpur for the DSC analysis.

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Authors and Affiliations

  1. Division of Bioprospection and Product Development, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O.-CIMAP, Lucknow, 226015, Uttar Pradesh, India

    Kuldeep Singh Yadav, Nidhi Srivastava, Vineet Kumar Rai, Pooja Rani Mina, Debabrata Chanda, Mahendra Pandurang Darokar & Narayan Prasad Yadav

  2. Division of Phytochemistry, CSIR-Central Institute of Medicinal and Aromatic Plants, P.O.-CIMAP, Lucknow, 226015, Uttar Pradesh, India

    Ranjana, Sudeep Tandon, P. V. Ajayakumar & Karuna Shanker

  3. Division of Toxicology and Experimental Medicine, CSIR-Central Drug Research Institute, Lucknow, 226031, Uttar Pradesh, India

    Navodayam Kalleti & Srikanta Kumar Rath

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  1. Kuldeep Singh Yadav
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Correspondence to Narayan Prasad Yadav.

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Yadav, K.S., Srivastava, N., Rai, V.K. et al. Cliv-92-Loaded Glycyrrhetinic Acid-Modified Chitosan Nanoparticles for Enhanced Hepatoprotection–Preparation, Characterization, and In Vivo Evaluation. AAPS PharmSciTech 22, 259 (2021). https://doi.org/10.1208/s12249-021-02130-7

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