Quality Markers of Traditional Chinese Medicine:Concept,Progress,and Perspective

As the most important complementary medication against a variety of diseases,traditional Chinese medicines(TCMs)have been extensively applied over thousands of years.Current quality control of herbal medicines,however,is in great dispute.Unlike chemical drugs,which have clear and validated quality standards,the content of only one(or a few)compounds of many herbs and preparations is currently assessed as an indicator of quality,even though the assessed compound(s)is neither closely associated with the efficacy nor representative of the medicine as a whole.Based on the clinical use,compatibility of multiple component prescriptions,and manufacturing process of TCM,the new concept of a TCM quality marker that was proposed in previous work is discussed further here.In addition,practical technological approaches are described for the qualitative analysis and quantification of TCMs including herbs,processed products,and preparations,which lead to the discovery and identification of specific chemicals as quality markers and new quality control patterns.The progress that has been made in TCM quality control is also addressed.This work provides useful information for the quality control of herbal medicines in the future.

Development of an LC–MS/MS method for the quantitation of deoxyglycychloxazol in rat plasma and its application in pharmacokinetic study

Deoxyglycychloxazol （TY501） is a glycyrrhetinic acid derivative which exhibits high anti-inflammatory activity and reduced pseudoaldosteronism compared to glycyrrhetinic acid. In this study, a sensitive and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was established for the quantitation of TY501 in rat plasma. Plasma samples were treated by precipitating protein with methanol and supernatants were separated by a Symmetry C8 column with the mobile phase consisting of methanol and 10 mM ammonium formate （containing 0.1% of formic acid） （90：10, v/v）. The selected reaction monitoring （SRM） transitions were performed at m/z 647.4→191.2 for TY501 and m/z 473.3→143.3 for astragaloside aglycone （IS） in the positive ion mode with atmospheric pressure chemical ionization （APCI） source. Calibration curve was linear over the concentration range of 5-5000 ng/mL. The lower limit of quantification was 5 ng/mL. The mean recovery was over 88%. The intra- and inter-day precisions were lower than 6.0% and 12.8%, respectively, and the accuracy was within ± 1.3%. TY501 was stable under usual storage conditions and handling procedure. The validated method has been successfully applied to a pharmacokinetic study after oral administration of TY501 to rats at a dosage of 10 mg/kg.

Potential synergic mechanism of Wutou-Gancao herb-pair by inhibiting efflux transporter P-glycoprotein

Wutou-Gancao herb-pair is extensively used to attenuate the toxicity and enhance the efficacy of aconite.In this study,potential synergic mechanism of the herb pair was investigated by utilizing multiple ap-proaches.In silico and in vitro Caco-2 cell models were applied to study the potential binding mode of bioactive ingredients existing in liquorice with P-glycoprotein(P-gp),as well as the inhibition effects on P-gp.Additionally,anti-inflammatory activity of aconitine(AC)combined with active ingredients of liquorice,as well as pharmacokinetic patterns of AC after co-administration was investigated.Anti-inflammatory effect of AC(1 mg/kg)in rats was enhanced in combination with bioactive ingredients of liquorice(10 mg/kg).In the meanwhile,the exposure of AC in vivo was altered,in terms of Cmax and AUC.For instance,the Cmax and AUC were increased to 1.9 and 1.3 folds,respectively,when used in combination with liquiritigenin.The in silico study revealed the potential binding mode with outward facing conformation of P-gp.The resulting data obtained from transport of rhodamine-123(Rh-123)across Caco-2 cell monolayer further indicated that the function of P-gp was inhibited by chemicals in liquorice.The synergic effect was therefore proposed to be attributed to inhibition of P-gp by liquorice since AC has been demonstrated to be the substrate of P-gp.The resuls revealed that potential synergic mechanism of Wutou-Gancao herb-pair by inhibiting function of key efflux transporter P-gp to enhance the exposure of AC in systematic circulation,and further the anti-inflammatory effect,which helps clarify the compatibility rationale of these two herbs.

Novel assays for quality evaluation of XueBiJing:Quality variability of a Chinese herbal injection for sepsis management

XueBiJing is an intravenous five-herb injection used to treat sepsis in China.The study aimed to develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)-or liquid chromatography-ultraviolet(LC-UV)-based assay for quality evaluation of XueBiJing.Assay development involved identifying marker constituents to make the assay therapeutically relevant and building a reliable one-point calibrator for monitoring the various analytes in parallel.Nine marker constituents from the five herbs were selected based on XueBiJing's chemical composition,pharmacokinetics,and pharmacodynamics.A selectivity test(for“similarity of response”)was developed to identify and minimize interference by nontarget constituents.Then,an intercept test was developed to fulfill“linearity through zero”for each analyte(absolute ratio of intercept to C response,<2%).Using the newly developed assays,we analyzed samples from 33 batches of XueBiJing,manufactured over three years,and found small batch-to-batch variability in contents of the marker constituents(4.1%-14.8%),except for senkyunolide I(26.5%).

A time-released osmotic pump fabricated by compression-coated method: Formulation screen, mechanism research and pharmacokinetic study

In this investigation,time-released monolithic osmotic pump(TMOP)tablets containing diltiazem hydrochloride(DIL)were prepared on the basis of osmotic pumping mechanism.The developed dosage forms were coated by Kollidon®SR-Polyethylene Glycol(PEG)mixtures via compression-coated technology instead of spray-coating method to form the outer membrane.For more efficient formulation screening,a three-factor five-level central composite design(CCD)was introduced to explore the optimal TMOP formulation during the experiments.The in vitro tests showed that the optimized formulation of DIL-loaded TMOP had a lag time of 4 h and a following 20-h drug release at an approximate zero-order rate.Moreover,the releasemechanismwas proven based on osmotic pressure and its profile could be well simulated by a dynamic equation.After oral administration by beagle dogs,the comparison of parameters with the TMOP tablets and reference preparations show no significant differences for C_(max)(111.56±20.42,128.38±29.46 ng/ml)and AUC_(0-48 h)(1654.97±283.77,1625.10±313.58 ng h/ml)but show significant differences for T_(max)(13.00±1.16,4.00±0.82 h).These pharmacokinetic parameters were consistent with the dissolution tests that the TMOP tablets had turned out to prolong the lag time of DIL release.

Application of immune checkpoint targets in the anti-tumor novel drugs and traditional Chinese medicine development

Immune checkpoints are the crucial regulators of immune system and play essential roles in maintaining self-tolerance,preventing autoimmune responses,and minimizing tissue damage by regulating the duration and intensity of the immune response.Furthermore,immune checkpoints are usually overexpressed in cancer cells or noninvasive cells in tumor tissues and are capable of suppressing the antitumor response.Based on substantial physiological analyses as well as preclinical and clinical studies,checkpoint molecules have been evaluated as potential therapeutic targets for the treatment of multiple types of cancers.In the last few years,extensive evidence has supported the immunoregulatory effects of traditional Chinese medicines(TCMs).The main advantage of TCMs and natural medicine is that they usually contain multiple active components,which can act on multiple targets at the same time,resulting in additive or synergistic effects.The strong immune regulation function of traditional Chinese medicine on immune checkpoints has also been of great interest.For example,Astragalus membranaceus polysaccharides can induce anti-PD-1 antibody responses in animals,and these antibodies can overcome the exhaustion of immune cells under tumor immune evasion.Furthermore,many other TCM molecules could also be novel and effective drug candidates for the treatment of cancers.Therefore,it is essential to assess the application of immune checkpoints in the development of new drugs and TCMs.In this review,we focus on research progress in the field of immune checkpoints based on three topics:(1)immune checkpoint targets and pathways,(2)development of novel immune checkpoint-based drugs,and(3)application of immune checkpoints in the development of TCMs.

Rapid Analysis and Identification of Absorbed Components and Their Metabolites of Yuanhu Zhitong Dropping Pill in Rat Plasma and Brain Tissue Using UPLC-Q-TOF/MS with Multivariate Statistical Analysis

Objective To establish an effective approach for rapid and comprehensive analysis on the absorbed and metabolic components in rats after ig administration of Yuanhu Zhitong Dropping Pill（YHZT）. Methods Based on the combination of UPLC-Q-TOF/MS and multivariate statistical analysis, the absorbed prototype constituents and their metabolites in rat plasma were rapidly analyzed and identified, and the components absorbed into brain were further identified by comparing the extracted ion chromatograms（EICs） of control and brain tissue samples of dosed rats. Results A total of 38 YHZT-related xenobiotic compounds were detected and identified as the potential bioactive constituents in rat plasma, including 24 absorbed prototype constituents and 14 metabolites. In particular, of all prototype constituents, 14 were also detected in rat brain tissue, indicating that they could penetrate the blood-brain barrier and enter into brain. Conclusion An effective method is established and applied to analyze the potential bioactive constituents in YHZT, which provides a pathway to further investigate the pharmacological pattern and mechanism of YHZT.

In Silico Molecular Docking Study of Repensine and Bentysrepinine against HBV DNA Polymerase

Bentysrepinine （Y101 ）, a derivative of repensine, is a novel di-peptide structure isolated from Dichondra repens. In vitro and in vivo tests exhibited that bentysrepinine markedly inhibited DNA-HBV and cccDNA activities. The binding mode of Y101 and repensine with DNA polymerase was driven by hydrophobic interactions. This might provide novel recognition of inhibitory effect of Y1 01 against HBV, though its inhibition mechanism needs to be validated by bio-assay at cellular level and of polymerase activity. Preliminary docking study suggested that Y101 might be able to inhibit HIV inverse transcriptase, also have the potential to interact with DNA polymerase and HCV NS5B polymerase.

Pay attention to situation of SARS-CoV-2 and TCM advantages in treatment of novel coronavirus infection

Since the outbreak of the new coronavirus epidemic,novel coronavirus has infected nearly 100,000 people in more than 110 countries.How to face this new coronavirus epidemic outbreak is an important issue.Basic reproduction number(R0)is an important parameter in epidemiology;The basic reproduction number of an infection can be thought of as the expected number of cases directly generated by one case in a population where all individuals are susceptible to infection.Epidemiology dynamics is a mathematical model based on a susceptibility-infection-recovery epidemic model.Researchers analyzed the epidemiological benefits of different transmission rates for the establishment of effective strategy in prevention and control strategies for epidemic infectious diseases.In this review,the early use of TCM for light and ordinary patients,can rapidly improve symptoms,shorten hospitalization days and reduce severe cases transformed from light and normal.Many TCM formulas and products have wide application in treating infectious and non-infectious diseases.The TCM theoretical system of treating epidemic diseases with TCM and the treatment scheme of integrated Chinese and Western medicine have proved their effectiveness in clinical practice.TCM can cure COVID-19 pneumonia,and also shows that the role of TCM in blocking the progress of COVID-19 pneumonia.

Development and Application of Network Toxicology in Safety Research of Chinese Materia Medica

Network toxicology that is an important branch of the network pharmacology emerges on the basis of network biology. It refers to study on the toxicological features of a constructed network model which is used to analyze toxic substances and their interaction and regulation in biological systems, particularly investigate the toxic effects of drugs and/or compatibility of medicines on body, and clarify the mechanism of toxicity. Network toxicology currently develops rapidly in safety prediction of Chinese materia medica （CMM）. The application of network toxicology to safety and toxicology study on CMM is extremely beneficial to identify the toxic components and potential incompatibility of CMM. Since CMM is a complex system with multi-components, multi-targets, and multi-interactions, the network toxicology in safety prediction of CMM faces three great challenges, including integration studies of bioinformatics, innovation of methods, and tools and risk assessment in future development of the network toxicology in CMM research. In this paper, relevant database, approaches and tools that network toxicology utilized in the safety study of CMM were carefully reviewed. Based on the progress made, the scientific development and modernization of CMM will be greatly enhanced.

Effect of age on the pharmacokinetics of polymorphic nimodipine in rats after oral administration

The previous investigation has proved that their existed pharmacokinetic difference between the different crystal forms of the polymorphic drugs after oral administration.However,no systemic investigations have been made on the change of this pharmacokinetic difference,resulted either from the physiological or from the pathological factors.In this paper,we used polymorphic nimodipine(Nim) as a model drug and investigated the effect of age difference(2- and 9-month old) on the pharmacokinetics after oral delivery in rats.As the results shown,for L-form of Nim(L-Nim),the AUC0–24 hin 2-month-old rats was 343.68747.15 ng h/m L,which is 23.36% higher than that in 9-month-old rats.For H-form of Nim(H-Nim),the AUC0–24 hin 2-monthold rats was 140.91719.47 ng h/m L,which is 54.64% higher than that in 9-month-old rats.The AUC0–24 h ratio between H-Nim and L-Nim was 2.44 in 2-month-old rats and 3.06 in 9-month-old rats.Since age difference could result in unparallelled change of the absorption and bioavailability of the polymorphic drugs,the results in this experiment are of value for further investigation of crystal form selection in clinical trials and rational clinical application of the polymorphic drugs.

Effects of total iridoid glycosides of Picrorhiza scrophulariiflora against non-alcoholic steatohepatitis rats induced by high-fat and high-sugar diet through regulation of lipid metabolism

Objective:To investigate the therapeutic effect of total iridoid glycosides of Picrorhiza scrophulariiflora(TIGP)on non-alcoholic steatohepatitis(NASH).Methods:SD rats were fed with high-fat and high-sugar diet for 8 weeks to establish NASH.TIGP were given orally at doses of 20,40 and 80 mg/kg/d for 4 weeks.Triglycerides assay(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),aspartate aminotransferase(AST),alanine aminotransferase(ALT),fasting plasma glucose(FPG),fasting insulin(FINS),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),chemokine-1(MCP-1),leptin(LEP)in serum were tested.TG,TC,superoxide dismutase(SOD),malondialdehyde(MDA),and free fatty acid(FFA)in liver tissue were determined by colorimetric methods.Steatosis of hepatocytes and inflammation was performed by pathological examination.Results:The results showed that TIGP significantly decreased TC,TG and FFA in liver tissue,increased SOD activity,decreased MDA content,decreased serum levels of TG,TC,HDL-C/LDL-C,ALT,AST,GLU,HOMA-IR,TNF-αand LEP,and in addition,improved steatosis of liver cells compared to NASH.Conclusion:TIGP had anti-fatty liver effect against NASH rats induced by high-fat and high-sugar diet.Its mechanism was related to the regulation of lipid metabolism and reduction of insulin resistance,through inhibition of oxidative stress and inflammation.

Application and prospect of toxicity quality markers of Chinese materia medica based on metabolomics

In view of the problem that the quality evaluation indicators divorced from effectiveness and safety in Chinese materia medica（CMM）, Prof. Chang-xiao Liu et al have put forward a concept of quality markers（Q-markers） of CMM, which provided a new thought for improving the CMM quality control method. The toxicity Q-markers of CMM are described as follows, the early toxicity prediction of CMM, the discovery, formation and susceptibility research of toxicity Q-markers of CMM. Metabolomics is a systematic biology approach, closely related to the toxicity phenotype of organism, has unique advantages in toxicity Q-markers research. The toxicity Q-markers of CMM are explained from the following four aspects in this review： 1） the establishment of toxicity prediction model of CMM, which is based on metabolomics,pattern recognition, and other techniques; 2） the discovery of toxicity Q-markers of CMM based on metabolomic, which is meaning that the toxicity Q-markers are interpreted regarding to the chemical composition analysis of CMM, preliminary screening and verification of the toxicity Q-markers and toxicity mechanism research; 3） the research on the formation of toxicity Q-markers based on metabolomics,which is described from biosynthetic pathway analysis and the secondary metabolites analysis of toxicity Q-markers; and 4） the susceptibility study of toxicity Q-markers based on metabolomics, namely, the key targets, and metabolic pathways of toxicity Q-markers are explained according to the target organ toxicity susceptibility caused by CMM, and then the toxicity susceptibility mechanism of CMM is clarified. Therefore, the application of metabolomics helps to systematically and comprehensively discover the quality control indicators related to the CMM safety, which further provides a rational evidence for the clinical application of toxicity of CMM.

Anti-mutagenicity of Swertiamarin and Its Metabolite in Incubated System of Human Intestinal Flora

Objective To study the biotransformation regulation and pharmacological effect of swertiamarin and its metabolite in incubated system of human intestinal flora. Methods Incubated system of human intestinal flora was utilized to research the intestinal metabolism of swertiamarin. Furthermore, mutagenic test and anti-mutagenic test were carried out to research the activity relationship of swertiamarin and its metabolite. Results Gentianine was found in the metabolites of swertiamarin. The pharmacological experiment indicated that swertiamarin and its metabolite both had good anti-mutagenic effect. Conclusion Swertiamarin is partly metabolized to gentianine after oral administration. They show similar anti-mutagenicity effects.

Investigation of the metabolites of five major constituents from Berberis amurensis in normal and pseudo germ-free rats

Berberis amurensis(Berberidaceae)is a traditional Chinese medicine,which is often used to treat hypertension,inflammation,dysentery and enteritis.It contains alkaloids,mainly including berberine,berbamine,magnoflorine,jatrorrhizine and palmatine.Berberis amurensis extracts(BAEs)is often orally taken.Oral herbs might be metabolized by intestinal bacteria in the small intestine.However,the interaction between the herb and the gut microbiota is still unknown.In the current study,UPLC/Q-TOFMS/MS combined with Metabolitepilot and Peakview software was used to identify the metabolites of BAEs in anti-biotic cocktail induced pseudo germ-free rats and normal rats.As a result,a total of 46 metabolites in normal rats were detected and its main metabolic pathways include demethylation,dehydrogenation,methylation,hydroxylation,sulfation and glucuronidation.Only 29 metabolites existed in pseudo germ-free rats.Dehydrogenated metabolites(M29,M30,M34 and M36),methylated metabolites(M33,M41 and M46)and other metabolites were not detected in pseudo germ-free rats.The result implied that the intestinal bacteria have an influence on the metabolism of BAEs.Furthermore,this investigation might contribute to the understanding of the metabolism of BAEs,and further promote its clinical application.

Phytochemistry,Metabolism, and Metabolomics of Ginseng

Ginseng, as a medicinal plant, has been used for thousands of years in China, Korea, and Japan, and the study on ginseng is a hotspot in the research field as evidenced by about 7000 scientific papers in PUBMED. In recent decades, many ginseng studies focused on the metabolism and metabolomics of ginseng or its active ingredients using modern bioanalytical technologies. To date, more than 200 ginsenosides and non-saponin constituents have been isolated and identified. In the past decades, rapid development of analytical technologies has facilitated the advancement of ginseng research in many ways. In this review, we focus on the advances of ginseng research in chemistry, pharmacology, and metabolomics. We also provide the comments on the significance as well as challenges of metabolomics-based ginseng studies.

Research and Application of Adlay in Medicinal Field

The traditional Chinese medicine（TCM） adlay has been used as a dietary supplement to promote health and treat various ailments for thousands of years.The effective and safe ingredients of TCM could be used as sources for developing new drugs.This paper reviews the main research and application of adlay seed in medicinal field in the following aspects：botanical resource,ethnopharmacological function,chemical constituents,pharmacology and pharmacokinetics,safety evaluation and toxicity,and clinical application.We hope that the review could help researchers mine the scientific values of adlay seed,innovative drug design,provide the guidance for the application in clinical therapy,and enhance the academic level and clinical efficacy of adlay seed.

Significance of neutralizing antibodies in COVID-19 therapy:progress and prospect

SARS-CoV-2 infection is a serious threat to human life and health all over the world,and COVID-19 is a global epidemic caused by SARS-CoV-2 infection.SARS-CoV-2 is highly infectious,strange and variable.Therefore,the treatment of COVID-19 must be urgent and targeted.However,vaccines and currently used drugs generally do not have the above-mentioned characteristics.Although convalescent plasma of COVID-19 has shown a clinical application value in the emergency treatment of critical patients,it shows great limitations.All human recombinant multivalent neutralizing nano-antibodies may meet the deficiency of COVID-19 therapy.Gene engineering technologies have been used to develop specific neutralizing antibody(nAB)drugs for the treatment of COVID-19 worldwide.Some of the candidate nAB drugs have been entered the clinical trials and can be used for the therapy of COVID-19 shortly.In the present review,we studied and analyzed n ABs for the treatment of COVID-19 and the progress and prospect from the following five aspects:1)The biological and clinical characteristics of SARS-CoV-2 infection;2)The feasibility of plasma therapy for convalescents with COVID-19;3)The technical routes of developing n Ab drugs;4)The current status of developing global COVID-19 antibodies;5)The difficulties and clinical use.

Discovery and Development of Artemisinin and Related Compounds

Artemisinin is isolated from the plant Artemisia annua,sweet wormwood,an herb employed in traditional Chinese medicine.Prof.You-you Tu discovered artemisinin in the 1960 s,so she was awarded the 2015 Nobel Prize in Physiology or Medicine.Artemisinin and its semi-synthetic derivatives are a group of drugs that possess the most rapid action of all current drugs against Plasmodium falciparum malaria.In this review,the author investigated history on discovery of artemisinin,ethnopharmacology of Artemisia plants,chemistry and pharmacological activities of the relative compounds,and introduced Tu and other Chinese and world scientists＇ contribution,development of artemisinin and the related compounds and registered and marketed artemisinin drugs in China,UK,and USA.The author also recalled the studies on the mechanism of action of artemisinins and artemisinin combination therapies and summed up the resistance issues.In Current Recommendations and the Global Plan for Insecticide Resistance Management in Malaria Vectors（CPIRM）,that the WHO prevents the development and manages the spread of insecticide resistance is summarized in the technical basis for coordinated action against insecticide resistance：preserving the effectiveness of modern malaria vector control.Prof.Tu re-emphasized the artemisinin resistant on five principles to the WHO.She called on the world＇s scientists to pay attention to the study of drug resistance,and hopes scientists to contribute to break resistance of artemisinins.

A review of lignans from genus Kadsura and their spectrum characteristics

Kadsura belongs to the Schisandroideae subfamily of Magnoliaceae.Plants from genus Kadsura are widely distributed in the South and Southwest of China.The plants of the genus are widely used as folk medicine for a long time in history,with the functions of relieving pain,promoting 'qi' circulation,activating blood resolve stasis,and applications in the treatment of rheumatoid arthritis and gastroenteric disorders.Lignans are the primary characteristic constituents with various biological activities of plants from genus Kadsura.This paper summarized 81 lignans isolated from the plants of genus Kadsura over the past eight years(from 2014 to 2021),which belong to five types:dibenzocyclooctadienes,spirobenzofuranoid dibenzocyclooctadienes,aryltetralins,diarylbutanes and tetrahydrofurans.Each type of these lignans possess typical characteristics in proton magnetic resonance(^(1)H NMR) and carbon-13 nuclear magnetic resonance(^(13)C NMR) spectra,the NMR regularities of these types of lingans were summarized,which provided a useful reference for the structural analysis of lignans.The relationships between lignans and pharmacodynamics were also systematically analyzed,lignans were predicted to be the quality markers(Q-marker) of Kadsura genus.