Beijing HONGHUI Meditech Co., Ltd.

Stroke, a destructive and burdensome disease, continues to affect an increasing number of families. HH-A, also known as (S)-6-((3',5-diallyl-2,4'-dihydroxy-[1,1'-biphenyl]-3-yl)amino)-6-oxohexane-1,5-diaminium chloride, has attracted attention for its ability to increase the oxygen-carrying capacity of hemoglobin and strongly alleviates ischemic brain injury in rat models of hypoxia. Owing to its low recovery rate after radioactive drug extraction, HH-A is thought to covalently bind to rat plasma proteins. In this study, intact protein, peptide, and amino acid analyses were performed on plasma samples via deconvolution of the intact protein, protein modification via proteomics, and protein mass spectrometry. HH-A was found to covalently bind to Lys413 of rat albumin. Molecular docking technology was used to simulate the binding mode of HH-A to rat albumin, the amino group of lysine is most likely to bind to the ortho-phenolic hydroxyl group judging from the reaction distance and the number of hydrogen bonds. This study demonstrated for the first time that honokiol and its derivatives (HH-A) covalently bind to rat plasma proteins and that the formation of new α,β-unsaturated aldehyde and ketone moieties in the drug is the key to these binding interactions. This experiment explains the binding mechanism of traditional Chinese medicine honokiol with plasma albumin at the molecular level, support the clinical research and development for HH-A. SIGNIFICANCE STATEMENT: This study demonstrated that honokiol and its derivatives (a new drug HH-A, also known as (S)-6-((3',5-diallyl-2,4'-dihydroxy-[1,1'-biphenyl]-3-yl)amino)-6-oxohexane-1,5-diaminium chloride) covalently bind to rat plasma proteins via deconvolution of the intact protein, protein modification via proteomics, and protein mass spectrometry. This work supports the clinical research and development for HH-A.

Honokiol, a bioactive component found in Magnolia officinalis, has shown in protecting against ischemic stroke in animal models. However, its poor water solubility has limited its clinical applications. In this study, we introduced a hydrophilic building block on the aromatic ring of honokiol, resulting in the synthesis of four new compounds (HH-A, -B, -C and -D) with significantly improved water solubility. We then investigated the neuroprotective effects of these compounds in mouse and rat models of transient middle cerebral artery occlusion/reperfusion (tMCAO/R) brain injury. Among the compounds tested, HH-A, also known as (S)-6-((3',5-diallyl-2,4'-dihydroxy-[1,1'-biphenyl]-3-yl)amino)-6-oxohexane-1,5-diaminium chloride, showed the most promising results. HH-A was found to significantly reduced the infarct volume and brain edema in mice. It also outperformed the other three compounds and honokiol, even surpassing the effects of edaravone dexborneol. Additionally, HH-A demonstrated dose-dependent improvements in body weight, neurological deficits, and infarct volume. Further analysis in tMCAO/R rat model revealed that HH-A treatment led to significant upregulations of Nrf2 and HO-1 in the brain. HH-A also significantly reduced the expression of HNE, and exhibited anti-apoptotic effects by decreasing the expression of Bax and increasing the expression of Bcl-2. This was further supported by a decrease in the number of TUNEL positive cells. Taken together, the neuroprotective effects of HH-A may be attributed to its ability to target the Nrf2/HO-1 signaling pathway, leading to reduced oxidative stress and apoptosis in the brain. These findings suggest that HH-A has potential as a therapeutic agent for the treatment of ischemic stroke.

Lipids metabolism was involved in the process of many types of tumor and alkylglycerone phosphate synthase (AGPS) was considered implicated in tumor process. Benzyl isothiocyanate (BITC) showed the inhibitory effect of tumor and AGPS activity, therefore, we screened a group of small molecular compound based on BITC by computer-aid design targeting AGPS and the results showed that the derivants could suppress the proliferation, the expression of tumor related genes such as survivin and Bcl-2, and the level of ether lipids such as lysophosphatidic acid ether (LPAe) and platelet activating factor ether (PAFe); however, the activity of caspase-3/8 was improved in glioma U87MG and hepatic carcinoma HepG2 cells in vitro.