Predicting cerebral acetylcholine dynamics with huperzine A pharmacokinetics in blood via mPBPK-PD modeling

Abstract

Huperzine A (HupA) is a highly selective, reversible acetylcholinesterase (AChE) inhibitor that exhibits neuroprotective effects and is clinically used to manage benign memory decline. However, the specific relationship between the pharmacokinetic (PK) profile of HupA and cerebral acetylcholine (ACh) dynamics remains poorly characterized. Here, we characterize the PK-pharmacodynamic (PD) properties of HupA in rats under both physiological and pathological conditions. Following a single intramuscular injection, HupA exhibits a short half-life but rapid brain penetration, while multiple dosing significantly enhances its brain exposure. In a middle cerebral artery occlusion (MCAO) rat model, HupA demonstrates increased brain distribution. Furthermore, HupA elevates ACh concentrations across multiple brain regions, concurrently modulating several monoamine neurotransmitters. Using a minimal physiologically based pharmacokinetic-pharmacodynamic (mPBPK-PD) modeling approach, cerebral ACh dynamics were accurately predicted based on the pharmacokinetics of HupA in systemic circulation. The developed mPBPK-PD model exhibits robust predictive performance and holds potential for guiding the optimization of clinical dosing regimens and improving the therapeutic efficacy of HupA.

Keywords

Huperzine A / Acetylcholinesterase / Neuroprotection / Acetylcholine / Ischemic stroke / mPBPK-PD / MCAO model

Funding

This work was supported by the National Key Research and Development Program of China (No. 2024YFA1308200), the National Natural Science Foundation of China (Nos. 82274009 and 81973556).

Declaration of competing interest

These authors have no conflict of interest to declare.

Acknowledgments

We gratefully acknowledge the use of BioRender for the creation of the scientific illustrations in this study.

References

[1] Le TTM, Hoang ATH, Nguyen NP, et al. A novel huperzine A-producing endophytic fungus Fusarium sp. Rsp5.2 isolated from Huperzia serrate. Biotechnol Lett. 2020; 42(6):987-995. https://doi.org/10.1007/s10529-020-02836-x.

[2] Ferreira A, Rodrigues M, Fortuna A, et al. Huperzine A from Huperzia serrata: a review of its sources, chemistry, pharmacology and toxicology. Phytochem Rev. 2016; 15(1):51-85. https://doi.org/10.1007/s11101-014-9384-y.

[3] Luo ZW, Yin FC, Wang XB, et al. Progress in approved drugs from natural product resources. Chin J Nat Med. 2024; 22(3):195-211. https://doi.org/10.1016/s1875-5364(24)60582-0.

[4] Zhang RW, Tang XC, Han YY, et al. Drug evaluation of huperzine A in the treatment of senile memory disorders. Acta Pharmacol Sin. 1991; 12(3):250-252.

[5] Tsai SJ. Huperzine A, a versatile herb, for the treatment of Alzheimer’s disease. J Chin Med Assoc. 2019; 82(10):750-751. https://doi.org/10.1097/jcma.0000000000000151.

[6] Wang LS, Zhou J, Shao XM, et al. Huperzine A attenuates cognitive deficits and brain injury in neonatal rats after hypoxia-ischemia. Brain Res. 2002; 949(1-2):162-170. https://doi.org/10.1016/s0006-8993(02)02977-3.

[7] Hu Q, Zhang R, Dong X, et al. Huperzine A ameliorates neurological deficits after spontaneous subarachnoid hemorrhage through endothelial cell pyroptosis inhibition. Acta Biochim Biophys Sin. 2024; 56(4):645-656. https://doi.org/10.3724/abbs.2024037.

[8] Zhang HY, Zheng CY, Yan H, et al. Potential therapeutic targets of huperzine A for Alzheimer’s disease and vascular dementia. Chem Biol Interact. 2008; 175(1-3):396-402. https://doi.org/10.1016/j.cbi.2008.04.049.

[9] Damar U, Gersner R, Johnstone JT, et al. Huperzine A as a neuroprotective and antiepileptic drug: a review of preclinical research. Expert Rev Neurother. 2016; 16(6):671-680. https://doi.org/10.1080/14737175.2016.1175303.

[10] Du Y, Liang HY, Zhang LM, et al. Administration of Huperzine A exerts antidepressant-like activity in a rat model of post-stroke depression. Pharmacol Biochem Behav. 2017; 158:32-38. https://doi.org/10.1016/j.pbb.2017.06.002.

[11] Zhou LY, Chen D, Guo XR, et al. Intravitreal injection of Huperzine A promotes retinal ganglion cells survival and axonal regeneration after optic nerve crush. Front Cell Neurosci. 2023; 17:1145574. https://doi.org/10.3389/fncel.2023.1145574.

[12] Yang XY, Geng L, Li R, et al. Huperzine A-liposomes efficiently improve neural injury in the hippocampus of mice with chronic intermittent hypoxia. Int J Nanomed. 2023; 18:843-859. https://doi.org/10.2147/ijn.s393346.

[13] An JR, Zhao YS, Luo LF, et al. Huperzine A, reduces brain iron overload and alleviates cognitive deficit in mice exposed to chronic intermittent hypoxia. Life Sci. 2020; 250:117573. https://doi.org/10.1016/j.lfs.2020.117573.

[14] Liang YQ, Tang XC. Comparative studies of huperzine A, donepezil, and rivastigmine on brain acetylcholine, dopamine, norepinephrine, and 5-hydroxytryptamine levels in freely-moving rats. Acta Pharmacol Sin. 2006; 27(9):1127-1136. https://doi.org/10.1111/j.1745-7254.2006.00411.x.

[15] Li F, Hu RF, Wang B, et al. Self-microemulsifying drug delivery system for improving the bioavailability of huperzine A by lymphatic uptake. Acta Pharm Sin B. 2017; 7(3):353-360. https://doi.org/10.1016/j.apsb.2017.02.002.

[16] Zhang RH, Wang C, Shi T, et al. Pharmacokinetics of HupA-PLGA-NPs of different sizes in the mouse blood and brain determined by LC-MS/MS. Eur Rev Med Pharmacol Sci. 2022; 26(4):1183-1195. https://doi.org/10.26355/eurrev_202202_28111.

[17] Singh YP, Kumar H. A recent update on huprine and its hybrids as a potential multifunctional agent for the treatment of Alzheimer’s disease. Chem Biol Drug Des. 2024; 103(2):e14478. https://doi.org/10.1111/cbdd.14478.

[18] 王许杰, 章海燕. 石杉碱甲改善认知功能的临床研究进展[J]. 中国新药与临床杂志, 2012, 31(12):707-712.

[19] Xu WJ, Zhang HY. Progress in clinical studies of huperzine a on improvement of cognitive function. Chin J New Drugs Clin Remed. 2012; 31(12):707-712. https://doi.org/10.1007/s11783-011-0280-z.

[20] Guo XR, Wu YH, Wang QQ, et al. Huperzine A injection ameliorates motor and cognitive abnormalities via regulating multiple pathways in a murine model of Parkinson’s disease. Eur J Pharmacol. 2023; 956:175970. https://doi.org/10.1016/j.ejphar.2023.175970.

[21] Judd JM, Jasbi P, Winslow W, et al. Inflammation and the pathological progression of Alzheimer’s disease are associated with low circulating choline levels. Acta Neuropathol. 2023; 146(4):565-583. https://doi.org/10.1007/s00401-023-02616-7.

[22] Cao Y, Jusko WJ. Applications of minimal physiologically-based pharmacokinetic models. J Pharmacokinet Pharmacodyn. 2012; 39(6):711-723. https://doi.org/10.1007/s10928-012-9280-2.

[23] Ayyar VS, Lee JB, Wang W, et al. Minimal physiologically-based pharmacokinetic (mPBPK) metamodeling of target engagement in skin informs anti-IL17A drug development in psoriasis. Front Pharmacol. 2022; 13:862291. https://doi.org/10.3389/fphar.2022.862291.

[24] Krzyzanski W, Milad MA, Jobe AH, et al. Minimal physiologically-based hybrid model of pharmacokinetics in pregnant women: application to antenatal corticosteroids. CPT Pharmacom Syst Pharma. 2023; 12(5):668-680. https://doi.org/10.1002/psp4.12899.

[25] Ogawa Y, Tsugita S, Torii Y, et al. Microdialysis-integrated HPLC system with dual-electrode detection using track-etched membrane electrodes for in vivo monitoring of dopamine dynamics. J Chromatogr B. 2024; 1247:124318. https://doi.org/10.1016/j.jchromb.2024.124318.

[26] Halfhide C, Cammarano TL, Anderson KA, et al. Using microdialysis to monitor dopaminergic support of limb-use control following mesencephalic neurosphere transplantation in a rodent model of Parkinson’s Disease. Behav Brain Res. 2024; 471:115121. https://doi.org/10.1016/j.bbr.2024.115121.

[27] Longa EZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 1989; 20(1):84-91. https://doi.org/10.1161/01.str.20.1.84.

[28] Bederson JB, Pitts LH, Tsuji M, et al. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke. 1986; 17(3):472-476. https://doi.org/10.1161/01.str.17.3.472.

[29] Riglet F, Mentre F, Veyrat-Follet C, et al. Bayesian individual dynamic predictions with uncertainty of longitudinal biomarkers and risks of survival events in a joint modelling framework: a comparison between stan, monolix, and NONMEM. AAPS J. 2020; 22(2):50. https://doi.org/10.1208/s12248-019-0388-9.

[30] Sang L, Yuan Y, Zhou Y, et al. A quantitative systems pharmacology approach to predict the safe-equivalent dose of doxorubicin in patients with cardiovascular comorbidity. CPT Pharmacom Syst Pharma. 2021; 10(12):1512-1524. https://doi.org/10.1002/psp4.12719.

[31] Djerada Z, Feliu C, Cazaubon Y, et al. Population pharmacokinetic-pharmacodynamic modeling of ropivacaine in spinal anesthesia. Clin Pharmacokinet. 2018; 57(9):1135-1147. https://doi.org/10.1007/s40262-017-0617-2.

[32] Van Belle K, Sarre S, Ebinger G, et al. Brain, liver and blood distribution kinetics of carbamazepine and its metabolic interaction with clomipramine in rats: a quantitative microdialysis study. J Pharmacol Exp Ther. 1995; 272(3):1217-1222. https://doi.org/10.1016/s0022-3565(25)24549-9.

[33] Wang H, Tang XC. Anticholinesterase effects of huperzine A, E2020, and tacrine in rats. Acta Pharmacol Sin. 1998; 19(1):27-30.

[34] Li C, Shi S. Neuroprotective effect of huperzine a on d-galactose-induced hearing dysfunction. Ear Nose Throat J. 2021; 100(3_suppl):269S-276S. https://doi.org/10.1177/0145561319864570.

[35] Mak SH, Li WM, Fu HJ, et al. Promising tacrine/huperzine A-based dimeric acetylcholinesterase inhibitors for neurodegenerative disorders: From relieving symptoms to modifying diseases through multitarget. J Neurochem. 2021; 158(6):1381-1393. https://doi.org/10.1111/jnc.15379.

[36] Dineley KT, Pandya AA, Yakel JL. Nicotinic ACh receptors as therapeutic targets in CNS disorders. Trends Pharmacol Sci. 2015; 36(2):96-108. https://doi.org/10.1016/j.tips.2014.12.002.

[37] Jing M, Li YX, Zeng JZ, et al. An optimized acetylcholine sensor for monitoring in vivo cholinergic activity. Nat Methods. 2020; 17(11):1139-1146. https://doi.org/10.1038/s41592-020-0953-2.

[38] Tang CY, Chen T, Kapadnis S, et al. Neuropharmacokinetics of two investigational compounds in rats: divergent temporal profiles in the brain and cerebrospinal fluid. Biochem Pharmacol. 2014; 91(4):543-551. https://doi.org/10.1016/j.bcp.2014.07.023.

[39] Si JC, Chen X, Qi KR, et al. Shengmaisan combined with Liuwei Dihuang Decoction alleviates chronic intermittent hypoxia-induced cognitive impairment by activating the EPO/EPOR/JAK2 signaling pathway. Chin J Nat Med. 2024; 22(5):426-440. https://doi.org/10.1016/s1875-5364(24)60640-0.

[40] Wang D, Yan B, Wang A, et al. Tu-Xian Decoction ameliorates diabetic cognitive impairment by inhibiting DAPK-1. Chin J Nat Med. 2023; 21(12):950-960. https://doi.org/10.1016/s1875-5364(23)60428-5.

[41] Zhou JC, Fan QL, Cai XY, et al. Ginkgo biloba extract protects against depression-like behavior in mice through regulating gut microbial bile acid metabolism. Chin J Nat Med. 2023; 21(10):745-758. https://doi.org/10.1016/s1875-5364(23)60496-0.

[42] Coleman BR, Ratcliffe RH, Oguntayo SA, et al. [+]-Huperzine A treatment protects against N-methyl-D-aspartate-induced seizure/status epilepticus in rats. Chem Biol Interact. 2008; 175(1-3):387-395. https://doi.org/10.1016/j.cbi.2008.05.023.

[43] Lu HY, Lu L, Jiang M, et al. Ultrastructural mitochondria changes in perihematomal brain and neuroprotective effects of Huperzine A after acute intracerebral hemorrhage. Neuropsychiatr Dis Treat. 2015:2649. https://doi.org/10.2147/ndt.s92158.

[44] Lin PP, Li XN, Yuan F, et al. Evaluation of the in vitro and in vivo metabolic pathway and cytochrome P450 inhibition/induction profile of Huperzine A. Biochem Biophys Res Commun. 2016; 480(2):248-253. https://doi.org/10.1016/j.bbrc.2016.10.039.

[45] Wu SL, Gan J, Rao J, et al. Pharmacokinetics and tolerability of oral dosage forms of huperzine a in healthy Chinese male volunteers: a randomized, single dose, three-period, six-sequence crossover study. Curr Med Sci. 2017; 37(5):795-802. https://doi.org/10.1007/s11596-017-1807-8.

[46] Becchi S, Chieng B, Bradfield LA, et al. Cognitive effects of thalamostriatal degeneration are ameliorated by normalizing striatal cholinergic activity. Sci Adv. 2023; 9(25):eade8247. https://doi.org/10.1126/sciadv.ade8247.

[47] Valdés Hernández MDC, Case T, Chappell FM, et al. Association between striatal brain iron deposition, microbleeds and cognition 1 year after a minor ischaemic stroke. Int J Mol Sci. 2019; 20(6):1293. https://doi.org/10.3390/ijms20061293.

[48] Chen APF, Chen L, Shi KW, et al. Nigrostriatal dopamine modulates the striatal-amygdala pathway in auditory fear conditioning. Nat Commun. 2023; 14:7231. https://doi.org/10.1038/s41467-023-43066-9.

[49] Oddo M, Hutchinson PJ. Understanding and monitoring brain injury: the role of cerebral microdialysis. Intensive Care Med. 2018; 44(11):1945-1948. https://doi.org/10.1007/s00134-017-5031-6.

[50] Peerdeman SM, Girbes ARJ, Vandertop WP. Cerebral microdialysis as a new tool for neurometabolic monitoring. Intensive Care Med. 2000; 26(6):662-669. https://doi.org/10.1007/s001340051230.

[51] Eiden M, Christinat N, Chakrabarti A, et al. Discovery and validation of temporal patterns involved in human brain ketometabolism in cerebral microdialysis fluids of traumatic brain injury patients. eBioMedicine. 2019; 44:607-617. https://doi.org/10.1016/j.ebiom.2019.05.054.

[52] Wei CC, Kong YY, Hua X, et al. NAD replenishment with nicotinamide mononucleotide protects blood-brain barrier integrity and attenuates delayed tissue plasminogen activator-induced haemorrhagic transformation after cerebral ischaemia. British J Pharmacol. 2017; 174(21):3823-3836. https://doi.org/10.1111/bph.13979.

[53] Li WL, Shi JF, Yu ZY, et al. SLC22A17 as a cell death-linked regulator of tight junctions in cerebral ischemia. Stroke. 2024; 55(6):1650-1659. https://doi.org/10.1161/strokeaha.124.046736.

[54] Liu SM, Jia XP, Liu B, et al. Suppression of cerebral ischemia injury induced blood brain barrier breakdown by dexmedetomidine via promoting CCN1. Aging. 2024; 16(4):3750-3762. https://doi.org/10.18632/aging.205557.

[55] Gao HM, Chen H, Cui GY, et al.Damage mechanism and therapy progress of the blood-brain barrier after ischemic stroke. Cell Biosci. 2023; 13(1):196. https://doi.org/10.1186/s13578-023-01126-z.

[56] Wang XN, Liu YS, Sun YY, et al. Blood brain barrier breakdown was found in non-infarcted area after 2-h MCAO. J Neurol Sci. 2016; 363:63-68. https://doi.org/10.1016/j.jns.2016.02.035.

[57] Chen XY, Wan SF, Yao NN, et al. Inhibition of the immunoproteasome LMP2 ameliorates ischemia/hypoxia-induced blood-brain barrier injury through the Wnt/β-catenin signalling pathway. Mil Med Res. 2021; 8(1):62. https://doi.org/10.1186/s40779-021-00356-x.

[58] Lochhead JJ, Ronaldson PT, Davis TP. The role of oxidative stress in blood-brain barrier disruption during ischemic stroke: Antioxidants in clinical trials. Biochem Pharmacol. 2024; 228:116186. https://doi.org/10.1016/j.bcp.2024.116186.

[59] Wang HY, Wu M, Diao JL, et al. Huperzine A ameliorates obesity-related cognitive performance impairments involving neuronal insulin signaling pathway in mice. Acta Pharmacol Sin. 2020; 41(2):145-153. https://doi.org/10.1038/s41401-019-0257-1.

[60] Li JJ, Meng XG, Li F, et al. Huperzine A combined with hyperbaric oxygen on the effect on cognitive function and serum hypoxia-inducible factor-1α Level in elderly patients with vascular dementia. Am J Transl Res. 2021; 13(6):6897-6904.

[61] Li J, Wu HM, Zhou RL, et al.Huperzine A for Alzheimer's disease. In:. Cochrane Database Syst Rev. 2008;(2): CD005592. https://doi.org/10.1002/14651858.CD005592.pub2.

[62] Xing SH, Zhu CX, Zhang R, et al. Huperzine a in the treatment of Alzheimer’s disease and vascular dementia: a meta-analysis. Evid-Based Compl Alt. 2014; 2014:363985. https://doi.org/10.1155/2014/363985.

[63] Huang WP, Zhu SZ, Liu XQ, et al. Cholinergic anti-inflammatory pathway involves in the neuroprotective effect of huperzine A on sepsis-associated encephalopathy. Chin Crit Care Med. 2016; 28(5):450-454.

[64] Wang ZF, Wang J, Zhang HY, et al. Huperzine A exhibits anti-inflammatory and neuroprotective effects in a rat model of transient focal cerebral ischemia. J Neurochem. 2008; 106(4):1594-1603. https://doi.org/10.1111/j.1471-4159.2008.05504.x.

[65] Zhang C, Li MG, Xie W, et al. Administration of Huperzine A microspheres ameliorates myocardial ischemic injury via α7nAChR-dependent JAK2/STAT3 signaling pathway. Eur J Pharmacol. 2023; 940:175478. https://doi.org/10.1016/j.ejphar.2022.175478.

[66] Su JQ, Chen KS, Sang X, et al. Huperzine a ameliorates sepsis-induced acute lung injury by suppressing inflammation and oxidative stress via α7 nicotinic acetylcholine receptor. Int Immunopharmacol. 2024; 141:112907. https://doi.org/10.1016/j.intimp.2024.112907.

[67] Liu C, Cai X, Ritzau-Jost A, et al. An action potential initiation mechanism in distal axons for the control of dopamine release. Science. 2022; 375(6587):1378-1385. https://doi.org/10.1126/science.abn0532.

[68] Krok AC, Maltese M, Mistry P, et al. Intrinsic dopamine and acetylcholine dynamics in the striatum of mice. Nature. 2023; 621(7979):543-549. https://doi.org/10.1038/s41586-023-05995-9.

[69] Delaney J, Nathani S, Tan V, et al. Enhanced cognitive flexibility and phasic striatal dopamine dynamics in a mouse model of low striatal tonic dopamine. Neuropsychopharmacology. 2024; 49(10):1600-1608. https://doi.org/10.1038/s41386-024-01868-5.

[70] Scognamiglio S, Aljohani YM, Olson TT, et al. Restoration of norepinephrine release, cognitive performance, and dendritic spines by amphetamine in aged rat brain. Aging Cell. 2024; 23(4):e14087. https://doi.org/10.1111/acel.14087.

[71] Griem-Krey N, Klein AB, Clausen BH, et al. The GHB analogue HOCPCA improves deficits in cognition and sensorimotor function after MCAO via CaMKIIα. J Cereb Blood Flow Metab. 2023; 43(8):1419-1434. https://doi.org/10.1177/0271678x231167920.

[72] Sood A, Mehrotra A, Dhawan DK, et al. Neuroprotective effects of Withania somnifera on ischemic stroke are mediated via anti-inflammatory response and modulation of neurotransmitter levels. Neurochem Int. 2024; 180:105867. https://doi.org/10.1016/j.neuint.2024.105867.

[73] Harik SI, Yoshida S, Busto R, et al. Monoamine neurotransmitters in diffuse reversible forebrain ischemia and early recirculation: Increased dopaminergic activity. Neurology. 1986; 36(7):971. https://doi.org/10.1212/wnl.36.7.971.

[74] Zhan QY, Kong FY. Mechanisms associated with post-stroke depression and pharmacologic therapy. Front Neurol. 2023; 14:1274709. https://doi.org/10.3389/fneur.2023.1274709.