Inhibition of the Growth of Calcium Oxalate Monohydrate by Glycyrrhizic Acid

doi:10.16553/j.cnki.issn1000-985x.2025.0017

Abstract

Abstract: Calcium oxalate monohydrate （COM） is the main component of kidney stones. This study explores the inhibitory effects of glycyrrhizic acid （GlA）， a multifunctional natural compound， on the crystallization process of COM and elucidates its mechanism of action. The crystallization kinetics of COM were examined by a calcium ion selective electrode， the impact of GlA on the size and morphology of COM crystals was quantitatively assessed through optical microscopy and scanning electron microscopy. Additionally， molecular simulations were employed to calculate the adsorption energy of GlA molecules on the COM crystal surface. The findings indicate that GlA serves as a highly effective inhibitor of COM crystallization， with the degree of inhibition positively correlating with GlA concentration. This inhibition is attributed to the specific adsorption of GlA on the ｛021｝ and ｛12 $1 ˉ$ ｝surfaces of COM， which significantly reduces the crystal aspect ratio. During the COM crystallization process， GlA plays a dual role： it complexes with Ca²⁺ in solution， thereby reducing their effective concentration， and it adsorbs onto the crystal surface via COO^-_（COM）…Ca²⁺…^-OOC_（GlA） interactions， hindering further crystal growth. These findings provide new ideas and potential therapeutic strategies for the prevention and treatment of kidney stones.

Key words: calcium oxalate monohydrate; glycyrrhizic acid; crystallization kinetics; inhibition; specific adsorption; complexation; kidney stone

CLC Number:

REN Yaqi, WANG Lei, LI Huimin, QU Yaqian, HAN Peizhuo, TAO Xutang. Inhibition of the Growth of Calcium Oxalate Monohydrate by Glycyrrhizic Acid[J]. Journal of Synthetic Crystals, 2025, 54(6): 1005-1012.

Figures/Tables 9

Fig.1 Molecular structure of GlA

Fig.2 COM microscopy images， diagram of crystal habit and their XRD patterns in the presence of different concentrations of GlA.（a） XRD patterns；（b） diagram of COM crystal habit in the absence of GlA；（c）~（h） microscopic crystal morphology images of COM at GlA concentrations of 0， 10， 30， 50， 100 and 200 μg/mL， respectively

Fig.3 Quantitative statistics of the size and morphology of COM crystals at different concentrations of GlA.（a） Variation of （100） crystal area of COM with GlA concentration；（b） plot of the thickness of COM as a function of GlA concentration；（c） aspect ratio of COM as a function of GlA concentration

Fig.4 Relationship between the inhibition effect of COM crystallization and the concentration of GlA.（a） Calcium ISE measurements of COM crystallization in the presence of GlA at varying concentrations；（b） percentage inhibition of COM crystallization by GlA

Fig.5 Crystal growth rate of COM with GlA concentration.（a） Schematic diagram of COM single crystal （left） and contact cross dual crystal （right）；（b）~（e） COM microscope images at 0 h and microscope images with the addition of 0 and 200 μg/mL GlA for 3 h；（f） relative growth rates of COM along ［001］，［010］ and ［100］ at different GlA concentrations （data are reported as the average of more than 20 crystals from 3 separate batches， the error bar is equal to two standard deviations）

Table 1 Ability to complex Ca2+ at different concentrations of GlA

C_GlA/（μg·mL^-1）	0	10	30	50	100	200
Ca²⁺ consumption rate/（10^-6 min^-1）	0	0.211 9±0.053 5	0.573 6±0.103 7	0.704 8±0.119 3	0.884 0±0.185 2	0.949 3±0.175 7

Table 2 Adsorption energy of the GlA and COM crystal planes

Crystal plane	（100）	（010）	（021）	（12 $1 ¯$ ）
Adsorption energy/eV	-3.491 65	-2.525 35	-4.377 63	-2.954 66

Table 2 Adsorption energy of the GlA and COM crystal planes

Crystal plane	（100）	（010）	（021）	（12 $1 ¯$ ）
Adsorption energy/eV	-3.491 65	-2.525 35	-4.377 63	-2.954 66

Fig.6 Schematic diagram of the atomic arrangement of the （100），（010），（021） and （121ˉ） plane of COM crystal and their interaction with GlA

Fig.7 Schematic diagram of GlA's inhibition of COM crystallization

References 19

1	KHAN S R， PEARLE M S， ROBERTSON W G， et al. Kidney stones［J］. Nature Reviews Disease Primers， 2016， 2（1）： 16008-16030.
2	朱佐轩，李　斯，汤伟伟，等. 人体生物矿化与病态结晶研究进展［J］. 科学通报， 2023， 68（4）： 339-355.
	ZHU Z X， LI S， TANG W W， et al. Biomineralization and pathological crystallization in vivo ［J］. Chinese Science Bulletin， 2023， 68（4）： 339-355 （in Chinese）.
3	ANG A J S， SHARMA A A， SHARMA A. Nephrolithiasis： approach to diagnosis and management［J］. Indian Journal of Pediatrics， 2020， 87（9）： 716-725.
4	STANKOVIĆ A， ŠAFRANKO S， JURIŠIĆ K， et al. Investigation of system complexity and addition of vitamin C on calcium oxalate precipitation［J］. Chemical Papers， 2020， 74（10）： 3279-3291.
5	BOTTRILL O， GRAHAM A， BOON M， et al. The impact of sugars on kidney stone formation［J］. Crystal Growth and Design， 2023， 23（11）： 8415-8425.
6	SUN Y Y， LIANG J， ZHANG Z R， et al. Extraction， physicochemical properties， bioactivities and application of natural sweeteners： a review［J］. Food Chemistry， 2024， 457： 140103.
7	ZHAO Z W， CAI M H， WANG P， et al. Application of glycyrrhizic acid and glycyrrhetinic acid in the drug delivery system［J］. Journal of Drug Delivery Science and Technology， 2024， 97： 105754.
8	CHUNG J， GRANJA I， TAYLOR M G， et al. Molecular modifiers reveal a mechanism of pathological crystal growth inhibition［J］. Nature， 2016， 536（7617）： 446-450.
9	FARMANESH S， CHUNG J， CHANDRA D， et al. High-throughput platform for design and screening of peptides as inhibitors of calcium oxalate monohydrate crystallization［J］. Journal of Crystal Growth， 2013， 373： 13-19.
10	CHAIYARIT S， PHUANGKHAM S， THONGBOONKERD V. Quercetin inhibits calcium oxalate crystallization and growth but promotes crystal aggregation and invasion［J］. Current Research in Food Science， 2024， 8： 100650.
11	KÜHNE T D， IANNUZZI M， DEL BEN M， et al. CP2K： an electronic structure and molecular dynamics software package-quickstep： efficient and accurate electronic structure calculations［J］. Journal of Chemical Physics， 2020， 152（19）： 194103.
12	VANDEVONDELE J， KRACK M， MOHAMED F， et al. Quickstep： fast and accurate density functional calculations using a mixed Gaussian and plane waves approach［J］. Computer Physics Communications， 2005， 167（2）： 103-128.
13	VANDEVONDELE J， HUTTER J. Gaussian basis sets for accurate calculations on molecular systems in gas and condensed phases［J］. The Journal of Chemical Physics， 2007， 127（11）： 114105.
14	KRACK M. Pseudopotentials for H to Kr optimized for gradient-corrected exchange-correlation functionals［J］. Theoretical Chemistry Accounts， 2005， 114（1）： 145-152.
15	HARTWIGSEN C， GOEDECKER S， HUTTER J. Relativistic separable dual-space Gaussian pseudopotentials from H to Rn［J］. Physical Review B， 1998， 58（7）： 3641-3662.
16	PERDEW J P， BURKE K， ERNZERHOF M. Generalized gradient approximation made simple［J］. Physical Review Letters， 1996， 77（18）： 3865-3868. DOI PMID
17	GRIMME S， EHRLICH S， GOERIGK L. Effect of the damping function in dispersion corrected density functional theory［J］. Journal of Computational Chemistry， 2011， 32（7）： 1456-1465. DOI PMID
18	LU T， CHEN F W. Multiwfn： a multifunctional wavefunction analyzer［J］. Journal of Computational Chemistry， 2012， 33（5）： 580-592.
19	SU Y， LI S， LI X， et al. Tartronic acid as a potential inhibitor of pathological calcium oxalate crystallization［J］. Advanced Science， 2024， 11（21）： 2400642.

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