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As crude drugs are natural products, their quality may vary. However, the degradation of the active ingredients in the compositional changes that occur during processing and preparation also affects the medicinal properties of the Kampo formula, which uses herbal medicines; therefore, a detailed investigation of the effects of compositional changes during preparation is required. Plant constituents vary in content depending on the year of cultivation and the plant part; however, detailed studies have rarely been reported for some crude drugs. Liquid chromatography-nuclear magnetic resonance/mass spectrometry revealed the degradation process of saponins, which are unstable components of the crude drug "Achyranthes root." The presence of diterpenes unstable with respect to drying temperature in the leaves of the crude drug "Leonurus herb" was revealed and their structures were elucidated. At the examination stage of the degradation process of perillaldehyde, the characteristic aromatic component of Perilla herb, it was elucidated that some specimens contained a small amount of perillaldehyde and that they contained more α-asarone. A trend toward lower ephedrine content was observed toward the tip of the above-ground branching of the Ephedra herb. Multivariate analysis was also introduced into the quality assessment of crude drugs and was established as a tool to identify bioactive compounds using the component diversity of crude drugs and to elucidate component differences due to the cultivation environment. [ABSTRACT FROM AUTHOR]

Erycibe paniculata Roxb. Is an ethno medicinal climber belongs to family Convolvulaceae commonly called Unamkodi in Tamil and Putta palatige in Telugu. It is found in India, Himalayas and Andaman Islands, has many ethno medicinal properties. Survey of literature on pharmacognosy and phytochemistry has elucidated that there are several ethnomedicinal plants which have been systematically studied for their pharmacognostic and phytochemical profiles. Crude drugs of a large number of ethnomedicinal plants are not investigated scientifically. Keeping this scenario in mind, the present study is planned to investigate phytochemical profile and also antibacterial activity of leaf extracts of Erycibe paniculata Roxb. Solvent extraction is carried by soxhlet method using different solvents based on their polarity and their yield percentages are calculated. Acetone and n-hexane showed high yield percentages compared to other solvents. The phytochemical tests were performed using standard methods that were reported and antibacterial activity is done by agar well diffusion method. For antibacterial activity we used two gram positive methicillin resistant Staphylococcus aureus and Bacillus subtilis and two gram negative bacteria-Pseudomonas aeruginosa and Escherichia coli. Ethyl acetate has shown maximum zone of inhibition towards all the bacteria tested and least is shown by toluene. [ABSTRACT FROM AUTHOR]

Background: The rhizome of Atractylodes chinensis (Asteraceae), a crude drug of Japanese Kampo medicines, has been administered to patients with edema, nephrotic syndrome, and gastrointestinal disorders. Essential oils, such as sesquiterpenoids (e.g., ß-eudesmol and hinesol) and atractylodin, are rich in the rhizomes. Previously, we discovered that atractylodin, a polyacetylene compound, found in an ethyl acetate (EtOAc)-soluble fraction from a methanol extract of A. chinensis rhizomes, possessed marked anti-inflammatory activities. Oral administration of the EtOAc-soluble fraction reduced immunoglobulin A (IgA) deposition in the renal glomeruli of high immunoglobulin A (HIGA) mice, a model of human IgA nephropathy. An increased serum IgA forms immune complexes and causes the deposition on renal glomeruli, leading to inflammation by the complement-mediated pathway. Objective: The aim of this study is to identify the compounds that reduce IgA deposition in an EtOAc-soluble fraction of A. chinensis rhizomes. Methods: Metabolites in the serum of HIGA and control BALB/c mice were analyzed by gas chromatography-mass spectrometry. A standard diet including each compound was fed to HIGA and BALB/c mice for 20 weeks to evaluate the improvement of glomerular IgA deposition. Results: Metabolomic analysis of serum suggested that the HIGA mice exhibit a state near the early stage of chronic kidney disease, compared with the BALB/c mice. When mice were orally administered each hydrophobic compound of chinensis rhizomes, it was revealed that atractylodin, as well as ß-eudesmol and (+)-hinesol, efficiently inhibited glomerular IgA deposition. Furthermore, the renal levels of complement component 3 (C3) and proinflammatory cytokine mRNAs were decreased, when the hydrophobic compounds were orally administered to HIGA mice. Conclusion: Atractylodin, ß-eudesmol, and (+)-hinesol may inhibit glomerular IgA deposition, probably by attenuating complement-mediated injuries and suppressing proliferation of mesangial cells in the renal glomeruli. These compounds might improve the pathological findings of human IgA nephropathy. [ABSTRACT FROM AUTHOR]

Human norovirus (HuNoV) is a major cause of acute gastroenteritis and foodborne diseases worldwide with public health concern, yet no antiviral therapies have been developed. In this study, we aimed to screen crude drugs, which are components of Japanese traditional medicine, "Kampo" to see their effects on HuNoV infection using a reproducible HuNoV cultivation system, stem-cell derived human intestinal organoids/enteroids (HIOs). Among the 22 crude drugs tested, Ephedra herba significantly inhibited HuNoV infection in HIOs. A time-of-drug addition experiment suggested that this crude drug more preferentially targets post-entry step than entry step for the inhibition. To our knowledge, this is the first anti-HuNoV inhibitor screen targeting crude drugs, and Ephedra herba was identified as a novel inhibitor candidate that merits further study. • 22 crude drugs were tested for their ability to inhibit HuNoV infection in HIOs. • Ephedra herba was identified as a novel anti-HuNoV inhibitor candidate. • Ephedra herba preferentially targets post-entry step for the inhibition. [ABSTRACT FROM AUTHOR]

In this study, we developed a convenient and easy-to-use origin identification method for antler velvets based on a simple DNA extraction technique and single-stranded tag hybridization chromatographic printed-array strip (STH-PAS). The primer sets used to detect Cervus elaphus, Rangifer tarandus, and 12S rRNA did not engage in non-specific reactions such as primer dimer formation. In both the triplex and singleplex assays, the sensitivity was < 1 ng DNA. Moreover, Cervus elaphus DNA could be detected in OTC crude drug products. Although the detection sensitivity resulting from the simplified extraction was slightly lower than that obtained with extraction by conventional methods, the amount of DNA was sufficient even from a small sample. The choice of a triplex or singleplex assay will depend on the purpose of the test. For example, if it is important to determine whether the antler velvet is derived from Cervus elaphus or Rangifer tarandus, a triplex assay is appropriate. If it is necessary to explore whether antler velvet from Cervus elaphus is included in an OTC crude drug product, a singleplex assay using the Cervus elaphus primer set is informative. If it is necessary to explore whether powdered antler velvet includes counterfeit products (from Rangifer tarandus), a singleplex assay employing the Rangifer tarandus primer is appropriate. The singleplex assay detects minor components even at a 1,000:1 ratio. Our study thus demonstrated the utility of a method combining simple DNA extraction with STH-PAS for efficient identification of the origin of antler velvets. [ABSTRACT FROM AUTHOR]

Context The mammalian circadian clock system regulates physiological function. Crude drugs, containing Polygalae Radix, and Kampō, combining multiple crude drugs, have been used to treat various diseases, but few studies have focussed on the circadian clock.Objective We examine effective crude drugs, which cover at least one or two of Kampō, for the shortening effects on period length of clock gene expression rhythm, and reveal the mechanism of shortening effects.Materials and methods We prepared 40 crude drugs. In the in vitro experiments, we used mouse embryonic fibroblasts from PERIOD2::LUCIFERASE knock-in mice (background; C57BL/6J mice) to evaluate the effect of crude drugs on the period length of core clock gene, Per2, expression rhythm by chronic treatment (six days) with distilled water or crude drugs (100 μg/mL). In the in vivo experiments, we evaluated the free-running period length of C57BL/6J mice fed AIN-93M or AIN-93M supplemented with 1% crude drug (6 weeks) that shortened the period length of the PERIOD2::LUCIFERASE expression rhythm in the in vitro experiments.Results We found that Polygalae Radix (ED50: 24.01 μg/mL) had the most shortened PERIOD2::LUCIFERASE rhythm period length in 40 crude drugs and that the CaMKII pathway was involved in this effect. Moreover, long-term feeding with AIN-93M+Polygalae Radix slightly shortened the free-running period of the mouse locomotor activity rhythm.Discussion and conclusions Our results indicate that Polygalae Radix may be regarded as a new therapy for circadian rhythm disorder and that the CaMKII pathway may be regarded as a target pathway for circadian rhythm disorders.

Aim: Drug‐induced liver injury (DILI) is a severe and life‐threatening immune‐mediated adverse effect, occurring rarely among treated patients. We examined genomic biomarkers in the Japanese population that predict the onset of DILI after using a certain class of drugs, such as Kampo products (Japanese traditional medicines). Methods: A total of 287 patients diagnosed as DILI by hepatology specialists were recruited after written informed consent was obtained. A genome‐wide association analysis and human leukocyte antigen (HLA) typing in four digits were performed. Results: We found a significant association (p = 9.41 × 10−10) of rs146644517 (G > A) with Kampo product–related DILI. As this polymorphism is located in the HLA region, we evaluated the association of HLA types and found that 12 (63.2%) of 19 Kampo‐DILI patients contained HLA‐B*35:01, whereas only 15.2% were positive for this HLA among healthy volunteers. The odds ratio was 9.56 (95% confidence interval 3.75–24.46; p = 2.98 × 10−6, corrected p = 4.17 × 10−5), and it increased to 13.55 compared with the DILI patients not exposed to Kampo products. The individual crude drug components in the Kampo products, including Scutellaria root (ougon in Japanese), rhubarb (daiou), Gardenia fruit (sanshishi), and Glycyrrhiza (kanzou), were significantly associated with HLA‐B*35:01. Conclusions: HLA‐B*35:01 is a genetic risk factor and a potential predictive biomarker for Kampo‐induced DILI in the Japanese population. [ABSTRACT FROM AUTHOR]

The current study aimed to identify the crude drugs associated with drug-induced liver injury (DILI) in 148 Kampo medicines prescribed throughout Japan using the Japanese Adverse Drug Event Report (JADER) database, a large-scale spontaneous reporting system in Japan. First, we tabulated the number of DILI reports from the report-based dataset and the background information from the patient-based dataset. Thereafter, we combined the 126 crude drugs into 104 crude drug groups to examine multicollinearity. Finally, the reporting odds ratios (RORs), 95% confidence intervals, p values for Fisher's exact test, and number of reports were calculated for each crude group to identify those associated with DILI. Notably, the number of adverse event reports for DILI (63,955) exceeded that for interstitial lung disease (51,347), the most common adverse event. In total, 78 crude drug groups (90 crude drugs) were reported to have an ROR > 1, a p < 0.05, and ≥10 reported cases. Our results highlight DILI as an essential issue, given that it was among the most frequently reported adverse drug reactions. We were able to clearly identify the crude drugs associated with DILI, which could help manage adverse drug reactions attributed to Kampo medicines and crude drugs. [ABSTRACT FROM AUTHOR]

We evaluated whether MinION, an inexpensive portable sequencer, can be used to identify the origin of crude drugs derived from animals. Standard and nonstandard crude drugs with different species of origin were examined. In addition, standards mixed with nonstandard samples were used. As a target gene, cytochrome c oxidase I was amplified and sequenced. The fast mode results had a slightly lower match ratio than high-accuracy mode, but the animals of origin were correctly determined by BLAST for all samples. For antler velvet derived from Rangifer tarandus, even when the sequences were aligned based on Cervus elaphus, the animal of origin was determined correctly. Minor contents could be detected from mixtures of two animals, if the mixtures contained at least 19:1 mtDNA when the coverage allele-fraction threshold was 0.05. By contrast, in fast mode, two sequences could not be separated due to the low accuracy of the base-calling for each read. For fieldwork, the species of origin of crude drugs could be identified with only simple DNA extraction and library preparation. Therefore, MinION appears to be a convenient tool for identifying the origins of crude drugs derived from animals. [ABSTRACT FROM AUTHOR]

Abstract Background Whole chloroplast genome (cpDNA) sequence is becoming widely used in the phylogenetic studies of plant and species identification, but in most cases the cpDNA were acquired from silica gel dried fresh leaves. So far few reports have been available to describe cpDNA acquisition from crude drugs derived from plant materials, the DNA of which usually was seriously damaged during their processing. In this study, we retrieved cpDNA from the commonly used crude drug Eriobotryae Folium (Pipaye in Chinese, which is the dried leaves of Eriobotrya japonica, PPY) using genome skimming technique. Results We successfully recovered cpDNA sequences and rDNA sequences from the crude drug PPY, and bioinformatics analysis showed a high overall consistency between the cpDNA obtained from the crude drugs and fresh samples. In the ML tree, each species formed distinct monophyletic clades based on cpDNA sequence data, while the phylogenetic relationships between Eriobotrya species were poorly resolved based on ITS and ITS2. Conclusion Our results demonstrate that both cpDNA and ITS/ITS2 are effective for identifying PPY and its counterfeits derived from distantly related species (i.e. Dillenia turbinata and Magnolia grandiflora), but cpDNA is more effective for distinguishing the counterfeits derived from the close relatives of Eriobotrya japonica, suggesting the potential of genome skimming for retrieving cpDNA from crude drugs used in Traditional Chinese Medicine for their identification.

Poria, the dried sclerotium of Wolfiporia cocos, is a medicinal mushroom that is widely used in traditional Japanese medicine. The fruit body of W. cocos is rarely found in the natural environment in Japan, therefore an optimized technique for fruit body formation is essential for producing new strains through crossbreeding and for biological research. Here, we developed a cultivation technique for fruit body formation of W. cocos using three strains collected from different areas of Japan. When mycelia were cultured on sawdust-based medium after liquid medium culture, all strains successfully formed fruit bodies as a brown honeycomb-like structure. Furthermore, we analyzed single nucleotide polymorphisms of the three strains using the STE3-like pheromone receptor protein gene, STE3.2, and found a genetic marker for discriminating one strain from the others. The results are expected to promote extensive studies on crossbreeding and domestic production of W. cocos. [ABSTRACT FROM AUTHOR]

Several traditional Japanese Kampo formulas are known to have inhibitory effects on infections with viruses that cause respiratory symptoms. Although some herbs and their components have been reported to suppress SARS-CoV-2 replication in vitro , it is difficult to compare effective Kampo formulas because of the different methods used in studies. Thus, we carried out in vitro experiments on the suppression of SARS-CoV-2 infection by Kampo formulas and crude drugs used for the common cold to compare their suppressive effects on virus infection. After infecting VeroE6/TMPRSS2 cells with SARS-CoV-2, lysates of the Kampo formulas and crude drugs were added, and after 24 h, the infectious titer in the medium was measured by the TCID50 method. Maoto was the most effective among the Kampo formulas, and Ephedrae herba was the most effective among the constituent crude drugs. However, a comparison of the suppressive effects of Ephedrae herba and Kampo formulas containing Ephedrae herba showed that the suppressive effect on virus infection did not depend on the content of Ephedrae herba. Based on the results, we believe that the use of Maoto among Kampo formulas is suitable as a countermeasure against COVID-19. [ABSTRACT FROM AUTHOR]

Several traditional Japanese Kampo formulas are known to have inhibitory effects on infections with viruses that cause respiratory symptoms. Although some herbs and their components have been reported to suppress SARS-CoV-2 replication in vitro, it is difficult to compare effective Kampo formulas because of the different methods used in studies. Thus, we carried out in vitro experiments on the suppression of SARS-CoV-2 infection by Kampo formulas and crude drugs used for the common cold to compare their suppressive effects on virus infection. After infecting VeroE6/TMPRSS2 cells with SARS-CoV-2, lysates of the Kampo formulas and crude drugs were added, and after 24 h, the infectious titer in the medium was measured by the TCID50 method. Maoto was the most effective among the Kampo formulas, and Ephedrae herba was the most effective among the constituent crude drugs. However, a comparison of the suppressive effects of Ephedrae herba and Kampo formulas containing Ephedrae herba showed that the suppressive effect on virus infection did not depend on the content of Ephedrae herba. Based on the results, we believe that the use of Maoto among Kampo formulas is suitable as a countermeasure against COVID-19.

Kampo is a form of traditional Japanese medicine, and its therapeutic strategy has been validated empirically over millennia, mainly in Asia. Kampo therapy aims to holistically prevent and treat disease based on the specific diagnosis Sho (in Japanese), in contrast with modern medical treatment which focuses on a patient's affected parts and local conditions. The medicines formulated using crude drugs derived from natural sources (Kampo formulas) are prescribed for patients according to their specific Sho, and thus the Kampo medication system is very complex. However, our previous study strongly suggested that Kampo medication theory could be explained by chemometrics and informatic approaches [Okada et al. in J Nat Med 70:107–114, 2016]. Here, we studied a group of seven formulas with Bupleurum Root and Scutellaria Root as the principal crude drugs. First, decoctions of the formulas were prepared and their supernatants were analyzed by non-targeted direct infusion mass spectrometry (MS) and principal component analysis, which is a type of unsupervised machine learning. Next, supervised machine learning was used to perform partial least squares modeling of the MS data matrix trained on the patients' constitution of Excess, Deficiency, or Medium between these two states (EDM) in Sho. The results showed that the correlation between the chemical fingerprints obtained by MS analysis and EDM could be modeled well using this approach. This cheminformatics modeling approach successfully interpreted part of the complex Kampo medication system studied using the fingerprints of formulas obtained by MS analysis and was consistent with the predicted Sho. [ABSTRACT FROM AUTHOR]

Background: Whole chloroplast genome (cpDNA) sequence is becoming widely used in the phylogenetic studies of plant and species identification, but in most cases the cpDNA were acquired from silica gel dried fresh leaves. So far few reports have been available to describe cpDNA acquisition from crude drugs derived from plant materials, the DNA of which usually was seriously damaged during their processing. In this study, we retrieved cpDNA from the commonly used crude drug Eriobotryae Folium (Pipaye in Chinese, which is the dried leaves of Eriobotrya japonica, PPY) using genome skimming technique. Results: We successfully recovered cpDNA sequences and rDNA sequences from the crude drug PPY, and bioinformatics analysis showed a high overall consistency between the cpDNA obtained from the crude drugs and fresh samples. In the ML tree, each species formed distinct monophyletic clades based on cpDNA sequence data, while the phylogenetic relationships between Eriobotrya species were poorly resolved based on ITS and ITS2. Conclusion: Our results demonstrate that both cpDNA and ITS/ITS2 are effective for identifying PPY and its counterfeits derived from distantly related species (i.e. Dillenia turbinata and Magnolia grandiflora), but cpDNA is more effective for distinguishing the counterfeits derived from the close relatives of Eriobotrya japonica, suggesting the potential of genome skimming for retrieving cpDNA from crude drugs used in Traditional Chinese Medicine for their identification. [ABSTRACT FROM AUTHOR]

The current resurgence in crude drug research and its moorings in underpinning the importance of phytochemicals provide the cue to study plants implicated as remedies in traditional heath care assists. The two exotic trees Adansonia digitata and Crescentia cujete and the medicinal seed legume Caesalpinia bonduc known for long in regional and local traditions investigated here show the species to contain constituents such as glycosides, flavonoids, phytosterols, tannins, free aminoacids, fixed oils and fats in somatic tissues, fruit pulp and seeds which can possibly be used as medicaments. As preliminary report on histochemical aspects alongside observations made on the feasibility quelling and controlling certain common potential pathogens lends credence to age old belief and claim that the said species can be hired in an organ specific and case specific manner to treat ailments. This fortifies the significance of TM in dealing with infections and treating vexing health disorders. [ABSTRACT FROM AUTHOR]

The clinical trial to evaluate the estrogenic effects of the crude drug derived from dry powder of a phytoestrogen-rich Thai herb Pueraria mirifica (White Kwao Krua) in five female volunteers with menopausal symptoms showed that the crude drug clearly improved the signs and symptoms related to menopause such as, hot flushes, frustration, sleep disorder, skin dryness, high blood cholesterol, oligomenorrhoea and amenorrhoea, with no change in the blood cells, liver and kidney functions, as well as other physiological status after four months of treatment. In four volunteers, treatments were continued to complete the one-year test period with half the dose and was found to maintain their satisfied menopausal relief status. The crude drug dosage was administered at 200 mg daily for three weeks a month during the first four months of treatment and 200 mg every other day for 20 days per month for the remaining of eight months. These doses were effective and safe as phytoestrogen treatment of menopausal symptoms.

Objective: This study aimed to assess the effectiveness of Kampo-sommelier practice, an active learning program on crude drugs used in Kampo formulations. Methods: The participants were fourth-year Tokai University School of Medicine students as of 2017. Eighteen small teams attended a 20-minute Kampo-sommelier practice session and were provided 10 kinds of crude drugs (Licorice, Cinnamon, Ginger, etc.) in three forms, original, cut, and powdered, while blinded to the drugs. Each team was asked to distinguish each drug in terms of form, scent, flavor, and color with reference to described characteristics. The ability to match the names of the drugs with their descriptions was assessed in the participants one month later, and also in human science "A" and medicine "B" students, without prior education, and pharmacy "C" students, with professional education. Results: The 117 participants received an average score of 6.2 ± 2.4 (mean ± S.D.) out of 10, which was significantly higher than 3.4 ± 1.8 in 97 "A" students and 3.1 ± 2.4 in 85 "B" students and lower than 8.4 ± 2.1 in 135 "C" students (p < 0.05 for all). Conclusions: The effectiveness of this team-based learning approach is suggested by the significantly higher scores of the participants. [ABSTRACT FROM AUTHOR]

The World Health Organization (WHO) has declared the coronavirus disease 2019 (COVID‐19) caused by the novel coronavirus SARS‐CoV‐2 a pandemic. There is, however, no confirmed anti‐COVID‐19 therapeutic currently. In order to assist structure‐based discovery efforts for repurposing drugs against this disease, we constructed knowledge‐based models of SARS‐CoV‐2 proteins and compared the ligand molecules in the template structures with approved/experimental drugs and components of natural medicines. Our theoretical models suggest several drugs, such as carfilzomib, sinefungin, tecadenoson, and trabodenoson, that could be further investigated for their potential for treating COVID‐19. [ABSTRACT FROM AUTHOR]

Purpose: The research was to determine specific parameters (macroscopy and microscopy) and non-specific parameters (loss on drying, total ash content, acid insoluble ash content, water-soluble extractive matter, ethanol-soluble extractive matter), heavy metal contaminant (Pb, Hg, As, Cu) and microbial contaminant (Total Plate Count and Total Yeast Mold). Research also determined a total flavonoid of M. oleifera leaf. Methodology: The method carried out according to the procedure stated in the Materia Medika Indonesia 5th edition. Result test compared to the standard stated in Indonesian Herbal Pharmacopoeia 2nd edition. Methods to detection Contaminants were referred to WHO guidelines for assessing the quality of herbal medicines. The determination of total flavonoid was performed by spectroscopic method. Results: Crude drug of M. oleifera leaf meet the specific parameters (macroscopy and microscopy). Nonspecific parameters of Africa, India, Local M. oleifera leaf shows that loss on drying (8.06 ± 0.03; 8.89 ± 0.31; 7.56 ± 0.17) %, total ash content (8.64 ± 0.43; 10.64 ± 0.90; 15.31 ± 0.87) %, acid insoluble ash content (0.56 ± 0.08; 0.35 ± 0.01; 0.36 ± 0.06)%, water-soluble extractive matter (21.38 ± 1.39; 30.12 ± 2.06; 12.68 ± 1.12) %, ethanol-soluble extractive matter (39.37 ± 1.51; 27.74 ± 2.44; 27.09 ± 1.43)%, contaminant test including heavy metal contaminant shows that Pb, Cd, As and Hg were not to be detected. Microbial contaminant (Total Plate Count and Total Yeast Mold) under limits of WHO standart. Total flavonoid content of Africa, India, Local M. oleifera leaf were 8.12 ± 0.52 mg/100 mg QE; 10.69 ± 0.15 mg/100 mg QE dan 13.08 ± 0.08 mg/100 mg QE respectively. Conclusion: Moringa Leaves meet specific parameters (macroscopy and microscopy test). Non-specific parameter tests of Africa, India, Local Moringa Leaf shows that loss on drying, acid insoluble ash content, water-soluble extractive matter, ethanol-soluble extractive matter, contaminant test including heavy metal contaminant shows that (Pb, Cd, As and Hg) and microbial contaminant (Total Plate Count and Total Yeast Mold) meet the standard required, only total ash content did not meet the standard. Total Flavonoid content shows that the extract of M. oleifera from local (Indonesia) variety is higher than others. [ABSTRACT FROM AUTHOR]

Aim: Many kampo formulas have been developed by combining crude drugs in various ways, providing diverse medicinal effects. Similarity between kampo formulas is usually determined based on their compositions and medicinal effects, but the entire network structure of Kampo formulas has not been analyzed systematically. In this study, I investigated the associations between kampo formulas based on their compositions and indications in order to provide an overview of the complex network structure of kampo formulas. In addition, relationships between crude drugs and indications were also investigated in a similar manner. Methods: The list of the 294 over‐the‐counter kampo formulas defined by the Ministry of Health, Labour and Welfare, Japan was analyzed to construct networks of kampo formulas. Kampo formulas were connected by edges if their crude drug compositions or indications had significant overlap. A network of crude drugs and indications was also constructed by linking crude drugs and indications if they co‐occurred in significantly many kampo formulas. Results: Three groups related to keishito, shosaikoto, and rikkunshito were identified in the network of kampo formulas based on crude drug composition. Three groups related to the treatment of gastric disorders, women's diseases, and neuralgia were also identified in the network of kampo formulas based on indications. Major groups were also identified in the network of crude drugs and indications. Conclusion: The networks of kampo formulas as well as crude drugs and indications were successfully obtained, which may help to facilitate our understanding of kampo formulas and crude drugs. [ABSTRACT FROM AUTHOR]

Crude drugs and Kampo formulations derived from natural materials such as plants, animals, and minerals are multicomponent medicines that contain numerous chemical constituents. Quantitative determination of characteristic constituents for quality control is crucial for the standardization and quality assurance of natural medicines. Quantitative assays to determine marker compound contents are commonly performed using HPLC systems. In order to achieve accurate quantitative determination, it is essential to use standard materials with well-defined purities corresponding to the target analytes. Many marker compounds used as standard materials must be purified and isolated from natural products while ensuring sufficient purity. However, the composition of impurities in the standard material differs among different batches due to differences in the raw materials and their extraction, separation, and purification processes. Therefore, controlling the purity of standard materials derived from natural products is more complex than that of synthetic substances. Quantitative NMR (qNMR), which has become widely used as an absolute quantitative method for low-molecule organic compounds, makes it possible to solve these issues. qNMR has been introduced into the crude drug section of the Japanese Pharmacopoeia (JP) for evaluating the purity of standard materials used for the assay. This review outlines an example of quantitative determination using relative molar sensitivity (RMS) based on qNMR adopted in the JP and introduces the latest efforts toward the application of qNMR to standard materials used for crude drugs in this context.

The root of Angelica acutiloba Kitagawa is an important crude drug in Kampo medicines (traditional Japanese medicine). Chemical evaluation of crude drugs is crucial to ensuring the safety and efficacy of herbal medicine; however, there is currently no chemical standard for the A. acutiloba crude drug in Japanese pharmacopoeia. (E)-ferulic acid (FA) is an important active ingredient of Angelica spp., including A. sinensis (Oliv.) Diels, and has been suggested as a marker for quality evaluation of those crude drugs. However, it has been controversial whether FA is a reliable marker constituent of A. acutiloba. To achieve effective extraction of FA from A. acutiloba, we compared three different extraction methods: alkaline hydrolysis, ethanol extraction, and hexane extraction. FA levels in these extracts were assessed using high performance liquid chromatography (HPLC), and alkaline hydrolysis was found to be the most effective. Furthermore, in the hydrolysate, FA was distinctly identified by thin layer chromatography (TLC) analysis. These results provide useful information for the quality control of the A. acutiloba crude drug. [ABSTRACT FROM AUTHOR]

Dengue virus (DENV) is transmitted to humans by Aedes mosquitoes and is a public health issue worldwide. No antiviral drugs specific for treating dengue infection are currently available. To identify novel DENV inhibitors, we analyzed a library of 95 compounds and 120 extracts derived from crude drugs (herbal medicines). In the primary screening, A549 cells infected with DENV-1 were cultured in the presence of each compound and extract at a final concentration of 10 μM (compound) and 100 μg/mL (extract), and reduction of viral focus formation was assessed. Next, we eliminated compounds and extracts which were cytotoxic using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hirsutine, an indole alkaloid of Uncaria rhynchophylla, was identified as a potent anti-DENV compound exhibiting high efficacy and low cytotoxicity. Hirsutine showed antiviral activity against all DENV serotypes. Time-of-drug-addition and time-of-drug-elimination assays indicated that hirsutine inhibits the viral particle assembly, budding, or release step but not the viral translation and replication steps in the DENV lifecycle. A subgenomic replicon system was used to confirm that hirsutine does not restrict viral genome RNA replication. Hirsutine is a novel DENV inhibitor and potential candidate for treating dengue fever.