Ginkgo Leaf Research Articles

An advanced bidirectional reflectance factor (BRF) spectral approach for estimating flavonoid content in leaves of Ginkgo plantations

As a key phenolic pigment concentrated in the surface tissues of leaves, flavonoids (Flav) are the major bioactive ingredients in Ginkgo leaf extracts. Flav are also marked natural antioxidants and significant indicators of biotic and abiotic stresses, critical for determining cultivation quality and enhancing Flav yield. In particular, area-based Flav (Flavarea) is related to the shortwave-blue light interaction within leaves per unit leaf area, whereas mass-based Flav (Flavmass) is useful for the quantitative assessment of Flav yield. In order to accurately estimate the contents of Flavarea and Flavmass in leaves of Ginkgo plantations, in this study, we developed an advanced bidirectional reflectance factor (BRF) spectra-based approach by reducing the effects of specular reflection and enhancing the absorption signals of Flav (in the shortwave-blue region of spectrum), using a suite of new spectral indices (SIs) (i.e., flavonoid index (FI), modified flavonoid index (mFI) and double difference index (DD)) calculated from the leaf clip equipped spectrometers-collected data. The results demonstrated that most of the SIs derived from the developed BRF spectra-based approach obtained relatively high performance for Flav estimation by alleviating adverse effects of specular reflection to different extents (CV-R2 = 0.60–0.76). In specific, DDnir434,421 selected from DD-type indices performed (CV-R2 = 0.76 for Flavarea; CV-R2 = 0.69 for Flavmass) better than other indices. These findings represent marked potentials of the developed BRF spectra-based approach for non-destructively estimating leaf Flav content, as well as improving the understanding of the mechanisms of specular effects on Flav estimations in leaves of Ginkgo plantations.

Neuroprotective and antioxidant activities of different polarity parts of the extracts of the Ginkgo biloba leaf and Zingiber officinale rhizome from Yongzhou.

In order to make better use of the two local dominant plant resources of Ginkgo biloba and Zingiber officinale from Yongzhou in Hunan province, the in vitro neuroprotective and antioxidant activities of extracts from the G. biloba leaf and Z. officinale rhizome, and the correlation between these two kinds of activities, were analyzed. The in vivo effects of these two plant extracts on aged mice blood physiology and central neuron cell activity were then determined after continuous gavage with the best polarity part at different concentrations (2, 4, 8 mg/ml). The results showed that the cell survival rate and superoxide dismutase (SOD) activity of the induced injury central neurons increased, although the malondialdehyde (MDA) content decreased gradually with the extract concentrations increasing in a certain range. Different polarity parts performed differently, even though they had the same concentration, with G. biloba always performing better than the Z. officinale rhizome at the same concentration and polarity. The order of G. biloba extract from superior to inferior was ethanol, ethyl acetate, n-butanol, chloroform, water, and petroleum ether (except that the petroleum ether part performed slightly better than the water part at 0.4 and 0.5 mg/ml), while the order of Z. officinale rhizome extract from superior to inferior was ethanol, chloroform, n-butanol, ethyl acetate, water, and petroleum ether. These two plant extracts demonstrated good in vitro effect against oxygen free radicals; the scavenging rate of superoxide free radicals had a significant positive correlation with the cell survival rate. The in vivo central nerve cell activity and SOD, glutathione peroxidase (GSH-PX) activity in aged mice blood serum increased while the MDA content decreased with treatment with these two extracts (p < 0.05). There were no significant changes in the number of leukocytes, lymphocytes, red blood cells, hemoglobin content, blood urine nitrogen, uric acid, creatinine, and the enzyme activity of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) (p > 0.05). G. biloba had a better in vivo effect than Z. officinale rhizome even though their concentration and polarity part were same. These results could provide some references for better development of these two plant extracts from Yongzhou in the field of neuroprotection.

Facile biosynthesis of ZnO/attapulgite nanocomposite for enhanced antimicrobial performance

• ZnO/APT nanocomposite was synthesized by a facile green one-pot protocol. • APT assisted forming of small sized ZnO NPs by offering rich nucleation sites. • ZnO NPs were formed by a bioreduction process without further thermal treatment. • Nanocomposite exhibited enhanced antibacterial activity and low cytotoxicity. An attapulgite (APT) supported ZnO nanocomposite (ZnO/APT) was developed through a facile green protocol mediated by Ginkgo biloba leaf extract. ZnO nanoparticles in the 5–10 nm size range, which were much smaller than pure ZnO particles, were uniformly decorated on APT surface, leading to an enhanced antibacterial performance of ZnO/APT nanocomposite compared with pure ZnO and the mixture of ZnO and APT. Moreover, the nanocomposite showed low cytotoxicity on L-O2 due to the green process and low Zn content. The obtained nanocomposite will be applicable in the medical field due to its safety and good performance.

Simultaneous determination of ginkgolide A, B, C, bilobalide and rutin in rat plasma by LC-MS/MS and its application to a pharmacokinetic study

Abstract A reliable LC-MS/MS method for the determination of five bioactive constituents (bilobalide, BLL; ginkgolide A, GLA; ginkgolide B, GLB; ginkgolide C, GLC; rutin) of Ginkgo biloba leaf extracts (GBE) in rat plasma was established, fully validated and applied to an intragastric pharmacokinetic study of a preparation of GBE in rat. Samples were extracted with ethyl acetate. C18 column was selected as analytical column in this method. Mobile phase was water with 0.01% formic acid and acetonitrile. Quantification was performed in negative multiple-reaction monitoring mode. Matrix instability of terpene lactones was noticed and hydrochloric acid was used as a stabilizer. This method showed good precision and accuracy, recovery was reproducible and matrix effect was negligible. Among four terpene lactones, BLL had the highest exposure and the shortest terminal half-life, GLA and GLB had lower exposure and longer terminal half-life, the exposure of GLC was lowest and its terminal half-life was the maximum, and all of them showed rapid absorption. This study provides a reference for determination of terpene lactones and flavonol glycoside prototypes in GBE and offers pharmacokinetic data of flavonol glycoside prototype in GBE.

Study on Integrated Pharmacokinetics of the Component-Based Chinese Medicine of Ginkgo biloba Leaves Based on Nanocrystalline Solid Dispersion Technology

BackgroundTo improve the dissolution and bioavailability of the component-based Chinese medicine of Ginkgo biloba leaves (GBCCM), a novel nanocrystalline solid dispersion of GBCCM (GBCCM NC-SD) was first prepared.MethodsGBCCM mainly containing high pure flavonoid aglycones (FAs) and terpenoid lactones (TLs) was used as the model drug. PVP K30 and SDS were used as solubilizers, combined stabilizers and carriers, and GBCCM NC-SD was prepared by high-pressure homogenization combined with freeze-dryer. Morphology and crystal characteristic of GBCCM NC-SD were analyzed. The dissolution and bioavailability evaluation were performed to investigate the feasibility of GBCCM NC-SD by in vitro dissolution and in vivo integrated pharmacokinetic models.ResultsAfter homogenizing for 30 cycles under the pressure of 650 bar and freeze-drying, GBCCM NC-SD with uniform quality would be obtained. The particle size, PDI and zeta potential were found to be 335.9 ± 32.8 nm, 0.29 ± 0.02 and −28.4 ± 0.7 mV respectively. Based on charged aerosol detector (CAD) technology, a new chromatographic method for simultaneous detection of eight components in GBCCM was developed. In vitro drug release study showed that the cumulative dissolution of FAs and TLs in GBCCM NC-SD increased from 12.77% to 52.92% (P < 0.01) and 90.91% to 99.21% (P < 0.05) respectively. In comparison with physical mixture of GBCCM and stabilizer (PM), the integrated pharmacokinetics AUC0-t of FAs and TLs in GBCCM NC-SD were significantly increased (P < 0.05), and the T1/2 of TLs was also significantly prolonged (P < 0.05).ConclusionThis study demonstrated that novel GBCCM NC-SD was prepared using Polyvinylpyrrolidone K30 (PVP K30) and Sodium dodecyl sulfate (SDS) as a synergetic stabilizer and also provided a feasible way to improve the dissolution and oral bioavailability of poorly soluble candidate antihypertensive drugs.

Eco-friendly Ginkgo Leaf Extract as a Green Corrosion Inhibitor to Protect N80 Steel in 1 M HCl

Eco-friendly Ginkgo Leaf Extract as a Green Corrosion Inhibitor to Protect N80 Steel in 1 M HCl

Therapeutic Versus Preventative Use of Ginkgo biloba Extract (EGb 761) against Indomethacin-Induced Gastric Ulcer in Mice

The main bioactive constituents in the standardized Ginkgo biloba leaf extract (EGb 761) are the terpene lactones and flavonoid glycosides. EGb 761’s antioxidant and anti-inflammatory properties have previously been demonstrated. Indomethacin-induced gastric ulcers have a multifactorial etiology and represent a major restriction to its therapeutic utility. The underlying ulcerogenic process involves oxidative and inflammatory biomolecular insults. This study was performed to explore the curative and preventative benefits of EGb 761 in experimentally-induced ulcers. To develop gastric ulcers in mice, indomethacin (40 mg/kg) was administered orally. EGb 761 (200 mg/kg) was given by gavage for 7 days before (preventative) and after (therapeutic) indomethacin administration. The histological alterations and macroscopic mucosal lesions were assessed. In gastric tissue homogenates, malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NO), and inflammatory cytokines were measured. The expressions of cyclooxygenase-2 (COX-2), cytokines, and proliferating cell nuclear antigen (PCNA) in the stomach mucosa were also investigated. The ulcer index, histological alterations, gastric oxidants, and inflammatory biomarkers were all significantly increased by indomethacin. In stomach specimens, it increased COX-2 and PCNA expression. EGb 761 treatments, both prophylactic and therapeutic, resulted in significant reductions in ulcer lesions, nitrosative and oxidative damage, and inflammatory markers, along with the lowering of COX-2 and PCNA expressions. Furthermore, in the fight against stomach ulcers, EGb 761 treatment was found to be more efficient than prevention.

MiR-146b-5p/TRAF6 axis is essential for Ginkgo biloba L. extract GBE to attenuate LPS-induced neuroinflammation.

Background: Neuroinflammation plays a crucial role in the pathogenesis and progression of various neurodegenerative diseases, including Alzheimer’s disease. The Ginkgo biloba leaf extract (GBE) has been widely used to treat cerebral and peripheral blood circulation disorders. However, its potential targets and underlying mechanisms regarding neuroinflammation have not yet been characterized. Aims: The purpose of this study was to investigate and validate the anti-neuroinflammatory properties of GBE against lipopolysaccharide (LPS)-mediated inflammation and to determine the underlying molecular mechanisms. Methods: The effect of GBE on LPS-induced release of inflammatory cytokines was examined using ELISA and western blot assay. The effects of GBE on NF-κB binding activity and translocation were determined via luciferase, streptavidin-agarose pulldown, and immunofluorescence assays. The potential targets of GBE were screened from the GEO and microRNA databases and further identified via qPCR, luciferase, gene mutation, and western blot assays. Results: GBE significantly inhibited LPS-induced pro-inflammatory responses in BV-2 and U87 cells, with no obvious cytotoxicity. GBE significantly induced miR-146b-5p expression, which negatively regulated TRAF6 expression by targeting its 3′-UTR. Thus, due to TRAF6 suppression, GBE decreases the transcriptional activity of NF-κB and the expression of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and cyclooxygenase (COX)-2, and finally reverses LPS-induced neuroinflammation. Conclusion: Our study revealed the anti-neuroinflammatory mechanism of GBE through the miR-146b-5p/TRAF6 axis and provided a theoretical basis for its rational clinical application.

Flavonoids affect the endophytic bacterial community in Ginkgo biloba leaves with increasing altitude.

Altitude affects plant growth and metabolism, but the effect of altitude on plant endophytic microorganisms is still unclear. In this study, we selected 16 Ginkgo biloba trees to study the response of leaves’ endophytes to flavonoids and altitude (from 530 m to 1,310 m). HPLC results showed that flavonoids in Ginkgo biloba leaves increased by more than 150% with attitude rising from 530 m to 1,310 m, which revealed a positive correlation with altitude. Ginkgo biloba might regulate the increased flavonoids in leaves to resist the increasing light intensity. 16S rDNA sequencing results showed that the endophytic bacterial communities of Ginkgo biloba at different altitudes significantly differed. Ginkgo leaf endophytes’ alpha diversity decreased with increasing flavonoids content and altitude. The increased flavonoids might increase the environmental pressure on endophytes and affect the endophytic community in Ginkgo biloba leaves. The bacterial network in Ginkgo biloba leaves became more complex with increasing altitude, which might be one of the strategies of leaf endophytes to cope with increasing flavonoids. Metagenomes results predicted with PICRUSt showed that the abundance of flavonoid biosynthesis and photosynthesis genes were significantly decreased with the increase of flavonoid contents. High flavonoid content in leaves appeared to inhibit microbial flavonoid synthesis. Our findings indicate that altitude can modulate microbial community structure through regulating plant metabolites, which is important to uncovering the interaction of microbes, host and the environment.

Determination of Vitamins, Trace Elements, and Phytochemical Compounds in Ginkgo Biloba Leaves Extracts

Determination of Vitamins, Trace Elements, and Phytochemical Compounds in Ginkgo Biloba Leaves Extracts

Real-time droplet size analysis using laser micrometer as a process analytical technology tool for continuous dripping process.

Process analysis and monitoring during the manufacturing of the dripping pills are essential. However, research on developing sensor‐based technology or process analytical technology (PAT) tools to analyze and monitor the dripping process is minimal. The purpose of this work is to develop a fast and non‐destructive laser detection system for quantitative visualization of droplets, which involves detecting the size of the droplet and calculating the weight of the dripping pills during the dripping process. Several factors influencing the detection performance of the detection system and the detection system capability for quantitation of the pill weight were explored. The laser detection system accurately detects the weight of the dripping pills with the coefficients of determination (R2) higher than 0.99. It was also robust concerning the variation in critical process parameters and critical material attributes. Furthermore, the laser detection system was successfully applied to the production line of Ginkgo biloba leaf dripping pills to monitor the dripping pills weight. The proposed laser detection system can analyze and monitor the dripping process in dripping pill manufacturing with stable performance, high accuracy, and high efficiency.

Traditional Chinese Medicine Injections for Diabetic Retinopathy: A Systematic Review and Network Meta-Analysis of Randomized Controlled Trials.

Background: The aim of this study was to compare the efficacy of different injected Traditional Chinese Medicines in the treatment of diabetic retinopathy (DR) and to provide a reference for the selection of adjuvant therapy for DR. Content: Related literature in multiple biological databases and websites was searched up to April 15, 2022, without language and publication time restrictions. A Bayesian network meta-analysis was used to analyze the included studies. Summary: Compared with conventional treatment, the combined use of injected Traditional Chinese Medicines, including astragalus, danhong, Ginkgo biloba extract powder, ginkgo leaf extract and dipyridamole (GLED), ligustrazine (LIG), mailuoning, puerarin, safflower, shuxuetong, safflower yellow sodium chloride, and xueshuantong (XST), can significantly improve the clinical effectiveness in patients with DR, while LIG, XST, and GLED can improve vision. The strength of the evidence ranged from high to very low. Outlook: In patients with DR, the combination of multiple injected Traditional Chinese Medicines is more effective than conventional treatment; some of these medicines may also improve visual acuity. This study may provide a good resource and reference for the selection of adjuvant therapy for DR.

Beneficial effects of Ginkgo biloba leaf extract on inflammatory markers: A systematic review and meta-analysis of the clinical trials.

The beneficial effects of Ginkgo biloba on cardio-metabolic markers have been reported. However, its effect on inflammation is not assessed in any meta-analysis. We performed a systematic review of randomized controlled trials evaluating the effects of Ginkgo biloba leaf extract (GBLE) on serum C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels. A systematic search was performed on four databases, including PubMed, Scopus, Web of Science, and Google Scholar, up to October 2021. After screening, 17 trials met our inclusion criteria. Trials were of 1-24 weeks of duration and included 1,104 participants. In the meta-analysis, the weighted mean differences (WMD) in change for serum CRP were -1.5mg/L (95% CI: -2.16, -0.85, p< 0.001). Moreover, WMD for serum IL-6 and TNF-α were in favor of the GBLE compared to the placebo [(-16.86 pg/mL, 95% CI: -19.38, -14.34, p< 0.001); and (-4.19 pg/mL, 95%CI: -5.14, -3.23, p< 0.001), respectively]. Subgroup analysis showed that GBLE has a beneficial effect on serum CRP at the baseline levels≥3mg/L and doses<500 mg/day. This meta-analysis showed that the GBLE could reduce serum inflammatory markers. Therefore, this medicinal herb might be a possible strategy for inflammation control.

Effects of ginkgo leaf tablet on the pharmacokinetics of rosiglitazone in rats and its potential mechanism

Context Ginkgo leaf tablet (GLT), a traditional Chinese herbal formula, is often combined with rosiglitazone (ROS) for type 2 diabetes mellitus treatment. However, the drug-drug interaction between GLT and ROS remains unknown. Objective To investigate the effects of GLT on the pharmacokinetics of ROS and its potential mechanism. Materials and methods The pharmacokinetics of 10 mg/kg ROS with 100/200 mg/kg GLT as single-dose and 10-day multiple-dose administration were investigated in Sprague-Dawley rats. In vitro, the effects of GLT on the activity of CYP2C8 and CYP2C9 were determined in recombinant human yeast microsomes and rat liver microsomes with probe substrates. Results The t 1/2 of ROS increased from 2.14 ± 0.38 (control) to 2.79 ± 0.37 (100 mg/kg) and 3.26 ± 1.08 h (200 mg/kg) in the single-dose GLT administration. The AUC0-t (139.69 ± 45.46 vs. 84.58 ± 39.87 vs. 66.60 ± 15.90 h·μg/mL) and t 1/2 (2.75 ± 0.70 vs. 1.99 ± 0.44 vs. 1.68 ± 0.35 h) decreased significantly after multiple-dose GLT treatment. The IC50 values of quercetin, kaempferol, and isorhamnetin, GLT main constituents, were 9.32, 7.67, and 11.90 μmol/L for CYP2C8, and 27.31, 7.57, and 4.59 μmol/L for CYP2C9. The multiple-dose GLT increased rat CYP2C8 activity by 44% and 88%, respectively. Discussion and conclusions The metabolism of ROS is attenuated in the single dose of GLT by inhibiting CYP2C8 and CYP2C9 activity, and accelerated after the multiple-dose GLT treatment via inducing CYP2C8 activity in rats, indicating that the clinical dose of ROS should be adjusted when co-administrated with GLT.

Role of Ginkgo Biloba in Alzheimer Disease

Alzheimer’s disease is a deadly neurodegenerative disease, with a complex etiology with many potential drug goals. Ginkgo biloba leaf extract has shown beneficial effects in the treatment of memory impairment, cognitive function of daily living (ADL) edema, inflammation and free toxins associated with traumatic brain injury (TBI). Alzheimer's dementia Flavonoid glycosides may have antioxidant effects that may reduce endothelial cell damage due to free radical oxidation thereby reducing the development of atherosclerosis. In addition, extraction of ginkgo may play an important role in treating minor dementia. The purpose of this article is to review the role of Ginkgo Biloba in Alzheimer's disease(AD).

Adsorption properties and mechanism of ginkgo biloba leaf-based materials for Cd (II) in aqueous solution.

The cheap and efficient heavy metal adsorbents were developed with the forestry and agricultural residues. In this study, three kinds of adsorbents, original ginkgo leaf (GL), NaOH modified ginkgo leaf (NaOH-GL), and KMnO4-modified ginkgo leaf (KMnO4-GL), were prepared and used to adsorb Cd (II) in aqueous solution respectively. The effects of the concentration of Cd (II), absorption time, the dosage of absorbent, and pH of solution on the adsorption process were explored by adsorption experiments. The results showed that the maximum adsorption capacity of Cd (II) by GL, NaOH-GL, and KMnO4-GL was 10.20 mg/g, 39.99 mg/g, and 48.82 mg/g, respectively, under the conditions of room temperature, adsorbent dosage 1g/L, adsorption time 300 min, and pH 6.0. The adsorption of Cd (II) by the three adsorbents accorded with the pseudo-second-order kinetic model and Langmuir isothermal adsorption model, which indicated that the rate-limiting step in the adsorption process was chemical adsorption process and mainly monolayer adsorption. The reason why NaOH-GL and KMnO4-GL could effectively adsorb Cd (II) was that the surface of the modified adsorbent was rough and porous, the number of active groups on the surface increased, and Na and Mn elements could promote the precipitation of Cd (II). The mechanism analysis of KMnO4-GL, which had the best adsorption effect, showed that the adsorption mechanism of Cd (II) might be surface adsorption, ion exchange, and complexing precipitation.

Effect of Ginkgo biloba leaf extract in combination with vitamin C, E and D on Aluminum Chloride induced Alzheimer in rats.

Effect of Ginkgo biloba leaf extract in combination with vitamin C, E and D on Aluminum Chloride induced Alzheimer in rats.

Ginkgo Biloba Leaf Extract Improves an Innate Immune Response of Peripheral Blood Leukocytes of Alzheimer’s Disease Patients

Background: One of the main features of Alzheimer’s disease (AD) pathology is failure in innate immune response and chronic inflammation. Lack of effective AD treatment means that more attention is paid to alternative therapy and drugs of natural origin, such as extract of Ginkgo biloba (EGb). The purpose of this study was to investigate the effect of EGb on the mechanisms of innate immune response of peripheral blood leukocytes (PBLs) in AD patients. Methods: In AD patients and healthy-age matched controls, the effect of EGb on two of innate immune reactions, i.e., PBLs resistance to viral infection ex vivo and production of cytokines, namely TNF-α, IFN-γ, IL-1β, IL-10, IL-15, and IFN-α, were investigated. The influence of EGb on inflammatory-associated genes expression that regulate innate immune response to viral infection and cytokine production, namely IRF-3, IRF-7, tetherin, SOCS1, SOCS3, NFKB1, p65, and MxA was also examined. Results: A beneficial effect of EGb especially in AD women was observed. EGb decreased production of TNF-α, IFN-γ, and IL-10 and increased IL-15 and IL-1β. The effect was more pronouncement in AD group. EGb also downregulated expression of investigated genes. Conclusions: EGb may have an advantageous properties for health management in elderly and AD sufferers but especially in women with AD. Improving peripheral innate immune cells’ activity by adding EGb as accompanying treatment in AD may be, in the long term, a good course to modify the disease progression.

Hydrothermal carbonization of waste ginkgo leaf residues for solid biofuel production: Hydrochar characterization and its pelletization

This study explored the potential of hydrothermal carbonization (HTC) for the production of a carbon-rich solid fuel from Ginkgo leaf residues (GLRs). Density, durability, hardness, hydrophobicity, and energy input estimates for HTC treated and untreated GLRs pellets were determined. Results showed that hydrothermal treatment significantly improved the physical properties of pellets made from GLRs, especially, at the temperature treatment of 220 °C. Pellet durability and hardness from samples treated at 220 °C (HT220) increased from 70.1% and 0.8 N/mm2 to 96.8% and 2.1 N/mm2 when compared to those made from untreated GLR (control). Hydrochar pellets had higher fixed carbon contents, elevated heating values, and enhanced energy densification ratio in comparison to the control pellets. The equilibrium moisture content decreased from 13.9% to 4.8% for HT220 pellets. The activation energy of HT220 pellets decreased from 44.6 to 26.8 kJ/mol in the first order reaction model and from 93.9 to 61.2 kJ/mol in the 3D diffusion model compared to the control pellets. The comprehensive combustibility index S was enhanced twice for HT220 pellets. The high durability, low moisture uptake, and improved pyrolysis property indicated that combining hydrothermal carbonization with pelletization is an efficient way for GLRs utilization as solid biofuel.

Efficacy and Safety of SID142 in Patients With Peripheral Arterial Disease: A Multicenter, Randomized, Double-Blind, Active-Controlled, Parallel-Group, Phase III Clinical Trial

Renexin® is a combination pill of cilostazol and Ginkgo biloba leaf extract that is used for the improvement of ischemic symptoms associated with peripheral arterial disease (PAD). SID142 is a controlled-release tablet of cilostazol (200 mg) and G biloba leaf extract (160 mg) that was developed to address the limitation of BID administration with Renexin. This study aimed to verify that SID142 was not inferior to Renexin in the treatment of patients with PAD. This was a multicenter, randomized, double-blind, active-controlled, parallel-group, Phase III clinical trial. Study subjects were randomized to receive SID142 once daily or Renexin twice a day for 12 weeks. The primary end point was a change in the patient assessment of lower leg pain intensity with the use of a visual analog scale (VAS) after 12 weeks of treatment. If the lower limit of the two-sided 95% CI was greater than -10, the study drug was declared noninferior to the reference drug. Secondary efficacy end points included cold sensation, ankle-brachial index, ankle systolic pressure, maximum walking distance, pain-free walking distance, and investigator's global assessment. Study group results were compared 4, 8, and 12 weeks after treatment. Adverse events were assessed as a safety end point. In total, 344 subjects from 19 medical centers were screened, and a total of 170 subjects were randomly assigned to either the SID142 (n=86) or the Renexin (n=84) group. Analysis of the change in lower extremity pain at 12 weeks compared with baseline revealed that SID142 was not inferior to Renexin (21.44 [19.23] vs 22.30 [17.75]; 95% CI, -7.70 to 5.97; P=0.5942). No significant differences were found between groups in any secondary efficacy end point. However, the incidence of adverse reactions was significantly lower in the SID142 group (22.35% vs 39.29%; P=0.0171). SID142 once daily was not inferior to Renexin twice a day for efficacy in patients with PAD. SID142 had a favorable safety profile. gov identifier: NCT03318276.