Pterocarpans and their biological activities: a review / 中国中药杂志

Pterocarpans, ubiquitous in Fabaceae, are protective active substances produced by the chemical defense system of plants. A total of 144 pterocarpans had been discovered before 2006. For the first time, we reported the 89 pterocarpans identified in 2006-2020. These pterocarpans not only demonstrate novel complex diversified genus-specific stereostructures but also display strong anti-microbial, anti-tumor, antioxidant, insecticidal, and anti-inflammatory activities. Through the projection of their biogenetic pathways and study of the pharmacological activities, the structure-activity correlation was further confirmed. The distribution of Leguminosae plants rich in pterocarpans has obvious regional characteristics. Therefore, the research and utilization of Leguminosae plant resources in China should be strengthened, and the popularity and application value of the geographical indicator plant resources should be improved. This paper serves as a reference for further research, development, and utilization of pterocarpans and their plant sources.

Metabolites and metabolic pathways of maackiain in rats based on UPLC-Q-TOF-MS / 中国中药杂志

Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to investigate the metabolites of maackiain in rats based on the prediction function of UNIFI data processing system and liver microsomal incubation in vitro. Ten metabolites of maackiain after oral absorption were reasonably deduced and characterized. It was found that the biotransformation of maackiain mainly included phase Ⅰ oxidation, dehydrogenation, phase Ⅱ sulfate conjugation, glucosylation conjugation, and glucuronic acid conjugation. Among them, the product of glucosylation conjugation, trifolirhizin, was identified by comparison with the reference for the first time. Liver microsomal incubation in vitro further confirmed the metabolites and metabolic pathways of maackiain in rats. The metabolites in the blood, urine, and feces complemented each other, which revealed the migration, metabolism, and excretion modes of maackiain in rats. This study lays a foundation for the further investigation of the metabolic mechanism of maackiain in vivo and the in-depth research on the mechanism of pharmacodynamics and toxicity.

Second-generation pterocarpanquinones: synthesis and antileishmanial activity

Despite the development of new therapies for leishmaniasis, among the 200 countries or territories reporting to the WHO, 87 were identified as endemic for Tegumentary Leishmaniasis and 75 as endemic for Visceral Leishmaniasis. The identification of antileishmanial drug candidates is essential to fill the drug discovery pipeline for leishmaniasis. In the hit molecule LQB-118 selected, the first generation of pterocarpanquinones was effective and safe against experimental visceral and cutaneous leishmaniasis via oral delivery. In this paper, we report the synthesis and antileishmanial activity of the second generation of pterocarpanoquinones. Methods: The second generation of pterocarpanquinones 2a-f was prepared through a palladium-catalyzed oxyarylation of dihydronaphtalen and chromens with iodolawsone, easily prepared by iodination of lawsone. The spectrum of antileishmanial activity was evaluated in promastigotes and intracellular amastigotes of L. amazonensis, L. braziliensis, and L. infantum. Toxicity was assessed in peritoneal macrophages and selective index calculated by CC50/IC50. Oxidative stress was measured by intracellular ROS levels and mitochondrial membrane potential in treated cells. Results: In this work, we answered two pertinent questions about the structure of the first-generation pterocarpanquinones: the configuration and positions of rings B (pyran) and C (furan) and the presence of oxygen in the B ring. When rings B and C are exchanged, we noted an improvement of the activity against promastigotes and amastigotes of L. amazonensis and promastigotes of L. infantum. As to the oxygen in ring B of the new generation, we observed that the oxygenated compound 2b is approximately twice as active against L. braziliensis promastigotes than its deoxy derivative 2a. Another modification that improved the activity was the addition of the methylenedioxy group. A variation in the susceptibility among species was evident in the clinically relevant form of the parasite, the intracellular amastigote. L. amazonensis was the species most susceptible to novel derivatives, whilst L. infantum was resistant to most of them. The pterocarpanoquinones (2b and 2c) that possess the oxygen atom in ring B showed induction of increased ROS production. Conclusions: The data presented indicate that the pterocarpanoquinones are promising compounds for the development of new leishmanicidal agents.(AU)

Estudo da regulação das DNA Estudo da regulação das DNA topoisomerases durante o desenvolvimento de resistência ao etoposídeo e do papel antitumoral de novos compostos em leucemias agudas / DNA topoisomerases's regulation study during resistance development toetoposide and antitumor role of new compounds in acute leukemias

Entre as neoplasias hematológicas, as leucemias agudas configuram o maior número de mortes a cada ano. A quimioterapia para estas neoplasias envolve inibidores de topoisomerase, como as antraciclinas, associados a outros fármacos. Entretanto, alguns pacientes não respondem ao tratamento devido ao desenvolvimento do fenótipo de resistência a múltiplas drogas (MDR), considerada a principal causa de refratariedade e falha no tratamento. Devido à alta taxa de proliferação celular, os tumores super expressam as DNA topoisomerases I e IIα humana (hTopo I e IIα), tornando essas enzimas bons alvos para o desenvolvimento de novos fármacos. Neste contexto, a procura por novos compostos capazes de aumentar a taxa de sobrevida global em pacientes com leucemias agudas e que sejam eficazes em células com fenótipo MDR se faz necessária. Os objetivos deste trabalho foram: a)desenvolver linhagens celulares de leucemias agudas resistentes ao etoposido (VP-16); b)caracterizar o fenótipo de resistência, mediado por alterações nas enzimas hTopo I e IIα; c)avaliar o mecanismo de ação dos novos compostos LQBs (LQB-118, -192, -223, -266, -268 e -326) e ácido pomólico (PA), como potenciais inibidores de hTopo I e/ou IIα; e d)investigar a atividade antitumoral dos compostos mais promissores nas linhagens de leucemias agudas resistentes em comparação às parentais. Foi demonstrado que dentre os compostos LQBs avaliados apenas LQB-118 e LQB-223 foram efetivos e específicos em inibir hTopoIIα. PA demonstrou ser um composto dual inibindo hTopo I e IIα. Os três compostos ativos não intercalam no DNA e atuam como inibidores catalíticos, não apresentando afinidade de interação ao sítio de ligação entre o DNA e camptotecina ou VP-16. Os estudos de modelagem molecular também sugeriram que LQB-118 e LQB-223 apresentam alta afinidade de ligação à região ATPase de hTopoIIα...

Acute leukemias represent the largest number of annual deaths from hematologic malignancy. The chemotherapy for these neoplasms involves topoisomerase inhibitors, such as anthracyclines, associated with other drugs. However, some patients do not respond to this treatment scheme because of the development of multiple drug resistance (MDR) phenotype.MDR phenotype is the main cause of refractoriness and treatment failure in acute leukemias. Due to the high rate of cell proliferation, tumors overexpress human DNA topoisomerases I and IIα (hTopo I and IIα), leading these enzymes as good targets for the development of new anticancer drugs. In this context, the searching for novel compounds capable of increase theoverall survival rate in patients with acute leukemias and be effective on cells with MDR phenotype is urgent. The objectives of this research are: a) todevelop acute leukemia cell linesresistant to etoposide (VP-16); b) to characterize the resistance phenotype mediated by changes on hTopo I and IIα; c) to evaluate themechanism of action of new compounds LQBs(LQB-118, -192, -223, -266, -268 and -326) and pomolic acid (PA), as potential inhibitors of hTopo I and/or IIα; and d) to investigate the antitumor activity of the most promising compounds on parental or resistant acute leukemia cell lines. It was demonstrated that among the LQBs evaluated only LQB-118 and LQB-223 were effective and specific to hTopo IIα and that PA inhibited both hTopo I and IIα. They did not intercalate into DNA and acted as catalytic inhibitors with poor affinity to interact with camptothecin or VP-16 biding site. The molecular modeling studies also suggested that LQB-118 and LQB-223 presented a high affinity to bindto ATPase region of hTopo IIα. Acute lymphoid CEM-R and myeloid leukemia U937-R cell lines were developed by exposition to increasing concentrations of VP-16...

Estudos pré-clínicos da pterocarpanoquinona LQB-118: toxicidade subaguda, eficácia em leishmaniose visceral experimental e mecanismo de ação / Preclinical studies of pterocarpanquinone lqb-118: subacute toxicity, efficacy in experimental visceral leishmaniasis and mechanism of action

A leishmaniose visceral (LV) é a forma mais severa de leishmaniose e é a segunda maior causa de mortes por doenças parasitarias depois da malária. O arsenal terapêutico contra a leishmaniose é pequeno, e cada um dos medicamentos disponíveis apresenta ao menos uma das desvantagens: toxicidade, eficácia, preço ou regime de tratamento. Nós temos concentrado esforços em estudar novos candidatos a fármacos como alternativas aos tratamentos atuais. A pterocarpanquinona LQB-118 foi desenhada e sintetizada com base em hibridação molecular e apresentou atividade em protozoários e linhagens celulares de leucemia. Resultados prévios demonstraram que a LQB-118 foi eficaz no tratamento da leishmaniose cutânea experimental e que o mecanismo de ação envolve a indução do estresse oxidativo com eventos característicos da morte celular por apoptose em Leishmania amazonensis. Neste estudo, foi observado que o tratamento com 10 mg/kg/dia por via oral de LQB- 118 inibiu o desenvolvimento de hepatoesplenomegalia em camundongos infectados com L. infantum, com uma redução de 99 por cento na carga parasitária. A análise toxicológica in vivo não apresentou nenhuma mudança nos parâmetros clínicos, bioquímicos ou hematológicos. A análise histológica evidenciou que os órgãos não apresentaram anormalidades, com a exceção do fígado, no qual foi observado focos de necrose com infiltração leucocitária com uma dose cinco vezes maior do que a dose terapêutica. Entretando, estas alterações não foram acompanhadas por aumento das transaminases. Para avaliar os eventos iniciais do mecanismo de ação da LQB- 118, promastigotas de L. amazonensis foram incubados com LQB-118 e antioxidantes. As perdas da viabilidade celular e do potencial de membrana mitocondrial não foram revertidas com os antioxidantes, embora a produção de EROs tenha sido prevenida, sugerindo que a produção de EROs não é a causa primária de morte do parasito...

Visceral leishmaniasis (VL) is the most severe form of leishmaniasis and is the second major cause of deaths byparasites after malaria. The arsenal of drugs against leishmaniasis is small, and each has a disadvantage in termsof toxicity, efficacy, price or treatment regimen. Our group has focused on studying new drug candidates asalternatives to current treatments. The pterocarpanquinone LQB-118 was designed and synthesized based onmolecular hybridization and exhibited antiprotozoal and anti-leukemic cell line activity. Our previous workdemonstrated that LQB-118 was an effective treatment of experimental cutaneous leishmaniasis and that themechanism of action involves induction of oxidative stress with characteristic events of cell death via apoptosisin Leishmania amazonensis. In this study, we observed that treatment with 10 mg/kg/day LQB-118 orallyinhibited the development of hepatosplenomegaly in L. infantum-infected mice, with a 99 percent reduction in parasiteload. The in vivo toxicological analysis showed no change in clinical, biochemical or hematological parameters.Histologically, all of the analyzed organs were normal with the exception of liver, in which focal points ofnecrosis with leukocyte infiltration were observed with a dose five times higher than the therapeutic dose.However, these changes were not accompanied by increase in transaminases. To assess the early effects of themechanism of action of LQB-118, promastigotes of L. amazonensis were incubated with LQB-118 andantioxidants. Cell viability and mitochondrial membrane potential were not reversed by the antioxidants,although the ROS production was, suggesting that ROS production is not the primary cause of parasite death...

Chemical constituents of Dalbergia odorifera / 中国中药杂志

Fourteen compounds were isolated from Dalbergia odoriferae and purified by repeated column chromatography on silica and sephadex LH-20 gel and structurally identified by spectral analysis. These compounds were identified as 4, 9-dimethoxy-3-hydroxypterocarpan (1), medicarpin (2), 2', 4', 5-trihydroxy-7-methoxyisoflavone (3), 2', 3', 7-trihydroxy-4'-methoxyisoflavan (4), formononetin (5), 3, 8-dihydroxy-9-methoxypterocarpan (6), koparin (7), 3-hydroxy-9-methoxypterocarp-6a-ene (8), 2'-hydroxyformononetin (9), stevenin (10), 2', 7-dihydroxy-4', 5'-dimethoxyisoflavone (11), lyoniresinol (12), 2, 4-dihydroxy-5-methoxy-benzophenone (13) and neokhriol A (14). Compounds 1, 3, 4, 6, 8, 12 and 14 were isolated from this plant for the first time. Antibacterial activity assay showed that compound 4 had inhibitory effect on Ralstonia solanacearum.

Simultaneous determination of 4 alkaloids and a flavonoid in Picrasmae Ramulus et Folium by RP-HPLC / 中国中药杂志

A RP-HPLC method was developed to evaluate the quality of Picrasmae Ramulus et Folium by simultaneous determination of five constituents including 1-hydroxymethyl-beta-carboline (1), 1-methoxicabony-beta-carboline (2), 4-methoxy-5-hydroxy-canthin-6-one (3), 4, 5-dimethoxy-canthin-6-one (4) and maackiain (5) in Picrasmae Ramulus et Folium. The samples were separated on a Kromasil RP-C18 (4.6 mm x 250 mm, 5 microm) column eluted with acetonitrile and 0.1% phosphoric acid as mobile phases in gradient mode. The detection wavelength was set at 254 nm. The calibration curves and linearity of the above five standards were determined as (1) Y = 6 525.6X + 37.25 (0.009-1.780 microg, r = 0.996 8), (2) Y = 3 662.3X + 41.55 (0.005-0.920 microg, r = 0.999 5), (3) Y = 3763.1X + 146.87 (0.015-3.060 microg, r = 0.999 0), (4) Y = 2 174.1X + 21.52 (0.003-0.620 microg, r = 0.999 5), and (5) Y = 276.25X + 7.65 (0.010-1.960 microg, r = 0.998 9), respectively. The method is simple and repeatable, and can be used for the quality assessment of Picrasmae Ramulus et Folium.

Glyceollins, a novel class of soybean phytoalexins, inhibit SCF-induced melanogenesis through attenuation of SCF/c-kit downstream signaling pathways

The anti-melanogenesis effect of glyceollins was examined by melanin synthesis, tyrosinase activity assay in zebrafish embryos and in B16F10 melanoma cells. When developing zebrafish embryos were treated with glyceollins, pigmentation of the embryos, melanin synthesis and tyrosinase activity were all decreased compared with control zebrafish embryos. In situ expression of a pigment cell-specific gene, Sox10, was dramatically decreased by glyceollin treatment in the neural tubes of the trunk region of the embryos. Stem cell factor (SCF)/c-kit signaling pathways as well as expression of microphthalmia-associated transcription factor (MITF) were determined by western blot analysis. Glyceollins inhibited melanin synthesis, as well as the expression and activity of tyrosinase induced by SCF, in a dose-dependent manner in B16F10 melanoma cells. Pretreatment of B16F10 cells with glyceollins dose-dependently inhibited SCF-induced c-kit and Akt phosphorylation. Glyceollins significantly impaired the expression and activity of MITF. An additional inhibitory function of glyceollins was to effectively downregulate intracellular cyclic AMP levels stimulated by SCF in B16F10 cells. Glyceollins have a depigmentation/whitening activity in vitro and in vivo, and that this effect may be due to the inhibition of SCF-induced c-kit and tyrosinase activity through the blockade of downstream signaling pathway.

Study on protective mechanism of kushenin injection on colonic mucosa of experimental colitis rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of NOD2 on colitis pathogenesis in experimental rats, and discuss therapeutical effect and mechanism of kushenin injection (OMT) on colitis in experimental rats.</p><p><b>METHOD</b>Fourty Sprague-Dawley (SD) rats were randomly divided into four groups: the normal control group, the model group, the SASP group, and the OMT group, with 10 rats in each group. Except the normal control group, models were established in the remaining three groups with TNBS. The OMT group was injected with kushenin injection, the SASP group was orally administered with mesalazine suspension, the model group and the normal group were orally administered with distilled water for 15 days. Colon lesion score and histological score of experimental rats were observed. Expression of NOD2, NF-kappaB p65 protein in rats colonic mucous was detected by immunohistochemistry. Expression of IL-6 in rat colon mucous was detected by ELISA.</p><p><b>RESULT</b>Compared with normal control group, the expression of NOD2, NF-kappaB p65 and IL-6 in colonic mucosa of the model group were significantly increased (P < 0.01). The SASP group and the OMT group showed lower expressions of NOD2, NF-kappaB p65 and IL-6 in colonic mucosa than the model group (P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>The over expression of colonic mucosa proteins NOD2 and NF-kappaB p65 and increasing secretion of IL-6 take part in the appearance and development of ulcerative colitis. OMT can attenuate ulcerative colitis and protect colonic mucosa by inhibiting expression of NOD2, NF-kappaB p65 and decreasing IL-6.</p>

Development and evaluation of a safe and effective sugar-free herbal lollipop that kills cavity-causing bacteria / 国际口腔科学杂志·英文版

Dental caries (tooth decay) is caused by a specific group of cariogenic bacteria, like Streptococcus mutans, which convert dietary sugars into acids that dissolve the mineral in tooth structure. Killing cariogenic bacteria is an effective way to control or prevent tooth decay. In a previous study, we discovered a novel compound (Glycyrrhizol A), from the extraction of licorice roots, with strong antimicrobial activity against cariogenic bacteria. In the current study, we developed a method to produce these specific herbal extracts in large quantities, and then used these extracts to develop a sugar-free lollipop that effectively kills cariogenic bacteria like Streptococcus mutans. Further studies showed that these sugar-free lollipops are safe and their antimicrobial activity is stable. Two pilot human studies indicate that a brief application of these lollipops (twice a day for ten days) led to a marked reduction of cariogenic bacteria in oral cavity among most human subjects tested. This herbal lollipop could be a novel tool to promote oral health through functional foods.

Chemical constituents of Ammopiptanthus mongolicus / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of aerial parts of Ammopiptanthus mongolicus.</p><p><b>METHOD</b>Isolation and purification were carried out on silica gel, Sephadex LH-20 and HPLC column chromatography. The structures of the compounds were identified by physico-chemical properties and spectral analysis.</p><p><b>RESULT</b>Nine compounds were isolated and identified as (+)-maackiain (1), brevifolin (2), 7-hydroxy-4'-methoxy isoflavanone (3), daidzein 4',7-diglucoside (4), genistein 4', 7-di-O-beta-D-glucoside (5), isolupalbigenin (6), ononin (7), beta-sitosterol (8), beta-daucosterol (9).</p><p><b>CONCLUSION</b>Compounds 2, 4 - 6 were obtained from the genus Ammopiptanthus for the first time.</p>

Application of kushenin on patients with chronic hepatitis C after renal transplantation / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of kushenin in treating patients with chronic hepatitis C after renal transplantation.</p><p><b>METHODS</b>Fifty-five patients were randomly assigned by lottery to the treatment group (29 cases) and control group (26 cases). The same immunosuppression therapy was given to all patients in both groups. Patients in the treatment group were treated with kushenin 0.6 g once a day, while those in the control group were treated with conventional liver protective agents such as vitamins. The treatment duration of both groups was 3 months. The incidences of serious hepatitis and acute rejection reaction, serum biochemistry parameters including indicators of liver and kidney functions, hepatic fibrosis index, and serum HCV-RNA were compared between the two groups.</p><p><b>RESULTS</b>(1) The incidence of serious hepatitis in the treatment group and the control group was 3.45% (1/29 cases) and 11.54% (3/26 cases), respectively, which was insignificantly different between the two groups (P=0.335). (2) The incidence of acute rejection in the treatment group was 6.90% (2/29 cases) and that in the control group was 7.69% (2/26 cases), showing insignificant difference (P=0.335). (3) The differences in serum alanine aminotransferase (ALT), direct bilirubin (DBIL), hyaluronic acid (HA), propeptide collagen type III (PC III), laminin (LN), collagen type IV (Col IV) levels between the two groups were insignificant before transplantation (P>0.05), while the above-mentioned parameters in the treatment group were significantly lower than those in the control group after transplantation (P<0.05). The difference in serum creatinine (SCr) and endogenous creatinine clearance rate (CCr) between the two groups was insignificant before and after transplantation (P>0.05). (4) The negative conversion rate of HCV-RNA in the treatment group was 31.03% (9/29 cases), significantly higher than the value of 11.54% (3/26 cases) in the control group after transplantation (P<0.05). (5) The levels of serum ALT and DBIL in patients with HCV-RNA converted to negative were significantly lower than those with still-positive HCV-RNA (P<0.05).</p><p><b>CONCLUSIONS</b>Kushenin has a certain effect on inhibiting the proliferation of HCV, protecting liver cells, and anti-liver fibrosis. On the other hand, it has no obvious influence on renal allograft function. Thus, the drug is clinically safe and effective for use in treating patients with chronic hepatitis C after renal transplantation.</p>

Studies on chemical constituents of Caragana spinifera / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the whole plant Caragana spinifera.</p><p><b>METHOD</b>The chemical constituents were isolated and repeatedly purified on silica gel column. The structures were elucidated by the NMR spectra and physico-chemical properties.</p><p><b>RESULT</b>Six compounds were isolated and identified as (6aR, 11aR) 4,9-dimethoxy-3-hydroxypterocarpan, (6aR,11aR)-4, 9-dihydroxy-3- methoxypterocarpan (melilotocarpane B), 5, 4'-dihydroxy-7-methoxyisoflavone, 7-hydroxy4'-methoxyisoflavone, 6, 7-dihydroxy4'-methoxyisoflavone, beta-sitosterol respectively.</p><p><b>CONCLUSION</b>All compounds were isolated from the plant for the first time.</p>

Propriedades anticâncer de pterocarpanos naturais / Anticancer properties of natural Pterocarpans

Os pterocarpanos apresentaram um núcleo tetracíclico derivado do núcleo fundamental das isoflavanonas. O presente trabalho teve como objetivos principais avaliar a atividade antitumoral "in vivo" de pterocarpanos e realizar estudos de mecanismo de ação. A avaliação da citotoxicidade "in vitro" mostrou que todas as células tumorais tratadas com 2,3,9-trimetoxipterocarpano foram inibidas, já a linhagem normal, não foi afetada. As células mononucleadas do sangue periférico também mostraram-se resistentes ao tratamento. A citotoxidade em células tumorais de dois derivados trimetoxilados de pterocarpanos, 3,9,10-trimetoxipterocarpano e 3,4,9-trimetoxipterocarpano também foi avaliada, e ambos não foram citotóxicos. Na tentativa de elucidar o mecanismo de ação citotóxica, a marcação para tubulina, actina, núcleo e lamina B foi realizada. As células MCF-7 tratadas com 2,3,9-trimetoxipterocarpano não apresentaram alteração nos microtúbulos da interfase e nem nos filamentos de actina, porém houve uma parada do ciclo celular na mitose em prometáfase. Grande número de células apresentou fusos mitóticos monopolares e outras células apresentavam fusos multipolares. A avaliação do conteúdo de DNA por citometria indicou que 2,3,9-trimetoxipterocarpano induz parada do ciclo em G2/M e que após 48 h de tratamento, além do bloqueio em G2/M o composto induz fragmentação do DNA...

Studies on chemical constituents in the root of Hedysarum polybotrys / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in the root of Hedysarum polybotrys Hand.-Mazz..</p><p><b>METHOD</b>The constituents were isolated by Sephadex LH-20 and silica gel column chromatography, and their structures were identified on the basis of spectral analysis.</p><p><b>RESULT</b>Five compounds, medicarpin (I), 3-hydroxy-9-methoxycoumestan (II), 3, 9-dihydroxycoumestan (III), beta-sitosterol (IV), beta-sitosterol-3-O-beta-D-glucoside (V) were obtained.</p><p><b>CONCLUSION</b>Compounds II, III, V were obtained from the plant for the first time.</p>