Cephaeline promotes ferroptosis by targeting NRF2 to exert anti-lung cancer efficacy.

CONTEXT: Cephaeline is a natural product isolated from ipecac (Cephaelis ipecacuanha [Brot.] A. Rich. [Rubiaceae]). It exhibits promising anti-lung cancer activity and ferroptosis induction may be a key mechanism for its anti-lung cancer effect. OBJECTIVES: This study investigates the anti-lung cancer activity and mechanisms of cephaeline both in vitro and in vivo. MATERIALS AND METHODS: H460 and A549 lung cancer cells were used. The cephaeline inhibition rate on lung cancer cells was detected via a Cell Counting Kit-8 assay after treatment with cephaeline for 24 h. Subsequently, the concentrations of 25, 50 and 100 nM were used for in vitro experiments. In addition, the antitumour effects of cephaeline (5, 10 mg/kg) in vivo were evaluated after 12 d of cephaeline treatment. RESULTS: Cephaeline showed significant inhibitory effects on lung cancer cells, and the IC50 of cephaeline on H460 and A549 at 24, 48 and 72 h were 88, 58 and 35 nM, respectively, for H460 cells and 89, 65 and 43 nM, respectively, for A549 cells. Meanwhile, we demonstrated that ferroptosis is the key mechanism of cephaeline against lung cancer. Finally, we found that cephaeline induced ferroptosis in lung cancer cells by targeting NRF2. DISCUSSION AND CONCLUSION: We demonstrated for the first time that cephaeline inhibits NRF2, leading to ferroptosis in lung cancer cells. These findings may contribute to the development of innovative therapeutics for lung cancer.

Alkaloids (emetine and cephalin) production - affected by full sunlight stress in Carapichea ipecacuanha.

This study evaluated the responses of Carapichea ipecacuanha to sunlight stress-induced changes in the electron transport chain and its extended effects on alkaloid production (emetine and cephalin). The treatments consisted of: (i). 50, 70, and 90% shading (controls) and their respective exposure to full sunlight; besides, full sunlight (55 days of direct sun exposure). Photosynthetic pigments, chlorophyll a fluorescence transient, antioxidant enzymatic system, and quantification of cephalin and emetine were analyzed. Several changes in the Chl a fluorescence induction were observed, such as a decline in the quantum yield of the conversion of photochemical energy and photosynthetic performance and; an increase in emetine production of plants exposed to full sunlight. These results demonstrated that ipecac plants are extremely sensitive to full exposure to solar radiation, especially in periods with high temperatures, such as in summer, however with increment in emetine production.

Emetine Is Not Ipecac: Considerations for Its Use as Treatment for SARS-CoV2.

Emetine is a potent antiviral that acts on many viruses in the low-nM range, with several studies in animals and humans demonstrating antiviral activity. Historically, emetine was used to treat patients with Spanish influenza, in the last stages of the pandemic in the early 1900s. Some of these patients were "black" with cyanosis. Emetine rapidly reversed the cyanosis and other symptoms of this disease in 12-24 h. However, emetine also has been shown to have anti-inflammatory properties and it appears it is these anti-inflammatory properties that were responsible for the effects seen in patients with Spanish influenza. Emetine, in the past, has also been used in 10s to 100s of millions of people at a dose of ~60 mg daily to treat amoebiasis. Based on viral inhibition data we can calculate a likely SARS-CoV2 antiviral dose of ~1/10th the amoebiasis dose, which should dramatically reduce the risk of any side effects. While there are no anti-inflammatory dose response data available, based on the potential mode of action, the anti-inflammatory actions may also occur at low doses. This paper also examines the toxicity of emetine seen in clinical practice and that seen in the laboratory, and discusses the methods of administration aimed at reducing side effects if higher doses were found to be necessary. While emetine is a "pure drug" as it is extracted from ipecac, some of the differences between emetine and ipecac are also discussed.

Emetine and cephaeline content in plants of Psychotria ipecacuanha in Costa Rica / Contenido de emetina y cefaelina en plantas de Psychotria ipecacuanha en Costa Rica / Emetina e conteúdo de cephaelina em plantas de Psychotria ipecacuanha na Costa Rica

Abstract The objective of this study was to identify the emetic metabolites in different parts of the P. ipecacuanha, a plant with emetic properties. Partial phytochemical analysis was performed to determine the presence of emetine and cephaeline in leaves, stems and roots. Both alkaloids were detected in the three plant parts analyzed. Highest alkaloid content was found in roots (8.55 mg/g), followed by stems (4.05 mg/g), and the lowest was found in leaves (2.4 mg/g). The cephaeline content (8.35 mg/g) was higher than that of emetine (6.65 mg/g) in all the three organs analyzed. Toxicity analysis of the crude extract showed a LD50 of 500 mg/kg.

Resumen El objetivo de este estudio fue identificar los metabolitos eméticos en diferentes partes de P.ipecacuanha, una planta con propiedades eméticas. Se realizó un análisis fitoquímico parcial, donde se determinó la presencia de emetina y cefalina en hojas, tallos y raíces. Ambos alcaloides se detectaron en las tres partes de la planta analizadas. El mayor contenido de alcaloides se encontró en las raíces (8.55 mg/g), seguido de los tallos (4.05 mg /g), y el más bajo se encontró en las hojas (2.4 mg/g). El contenido de cefalina (8,35 mg/g) fue mayor que el de emetina (6,65 mg/g) en los tres órganos analizados. El análisis de toxicidad del extracto crudo mostró una DL50 de 500 mg/kg.

Resumo O objetivo deste estudo foi identificar os metabólitos eméticos em diferentes partes do P. ipecacuanha, uma planta com propriedades eméticas. Foi realizada análise fitoquímica parcial para determinar a presença de emetina e cefelina nas folhas, caules e raízes. Ambos os alcalóides foram detectados nas três partes da planta analisadas. O maior teor de alcalóides foi encontrado nas raízes (8,55 mg/g), seguido dos caules (4,05 mg/g), e o menor foi encontrado nas folhas (2,4 mg/g). O conteúdo de cefhaelina (8,35 mg/g) foi superior ao de emetina (6,65 mg/g) nos três órgãos analisados. A análise de toxicidade do extrato bruto mostrou um LD50 de 500 mg/kg.

Emetine, Ipecac, Ipecac Alkaloids and Analogues as Potential Antiviral Agents for Coronaviruses.

The COVID-19 coronavirus is currently spreading around the globe with limited treatment options available. This article presents the rationale for potentially using old drugs (emetine, other ipecac alkaloids or analogues) that have been used to treat amoebiasis in the treatment of COVID-19. Emetine had amongst the lowest reported half-maximal effective concentration (EC50) from over 290 agents screened for the Middle East respiratory syndrome (MERS) and severe acute respiratory syndrome (SARS) coronaviruses. While EC50 concentrations of emetine are achievable in the blood, studies show that concentrations of emetine can be almost 300 times higher in the lungs. Furthermore, based on the relative EC50s of emetine towards the coronaviruses compared with Entamoeba histolytica, emetine could be much more effective as an anti-coronavirus agent than it is against amoebiasis. This paper also discusses the known side effects of emetine and related compounds, how those side effects can be managed, and the optimal method of administration for the potential treatment of COVID-19. Given the serious and immediate threat that the COVID-19 coronavirus poses, our long history with emetine and the likely ability of emetine to reach therapeutic concentrations within the lungs, ipecac, emetine, and other analogues should be considered as potential treatment options, especially if in vitro studies confirm viral sensitivity.

Cyclotides Isolated from an Ipecac Root Extract Antagonize the Corticotropin Releasing Factor Type 1 Receptor.

Cyclotides are plant derived, cystine-knot stabilized peptides characterized by their natural abundance, sequence variability and structural plasticity. They are abundantly expressed in Rubiaceae, Psychotrieae in particular. Previously the cyclotide kalata B7 was identified to modulate the human oxytocin and vasopressin G protein-coupled receptors (GPCRs), providing molecular validation of the plants' uterotonic properties and further establishing cyclotides as valuable source for GPCR ligand design. In this study we screened a cyclotide extract derived from the root powder of the South American medicinal plant ipecac (Carapichea ipecacuanha) for its GPCR modulating activity of the corticotropin-releasing factor type 1 receptor (CRF1R). We identified and characterized seven novel cyclotides. One cyclotide, caripe 8, isolated from the most active fraction, was further analyzed and found to antagonize the CRF1R. A nanomolar concentration of this cyclotide (260 nM) reduced CRF potency by ∼4.5-fold. In contrast, caripe 8 did not inhibit forskolin-, or vasopressin-stimulated cAMP responses at the vasopressin V2 receptor, suggesting a CRF1R-specific mode-of-action. These results in conjunction with our previous findings establish cyclotides as modulators of both classes A and B GPCRs. Given the diversity of cyclotides, our data point to other cyclotide-GPCR interactions as potentially important sources of drug-like molecules.

Pick your poison: what's new in poison control for the preschooler.

Accidental childhood poisonings are a major public health concern despite many efforts to alleviate this problem. While the rate of pediatric fatalities due to poisonings have decreased over the last two decades, poison control centers around the US have collectively fielded over one million calls with regard to toxic exposures in the preschool age group. According to the American Association of Poison Control Centers nearly half of all human exposures reported last year involved children under six. By focusing poison prevention efforts on the preschooler, we can attempt to decrease morbidity and mortality in the most vulnerable age group affected. Although the subject is still prevalent, current discussion on this topic is limited. Newer literature discusses past initiatives such as child resistant packaging and sticker deterrent programs and addresses their efficacy. This article revisits older mechanisms of prevention as well as the science behind the human motivation to change one's own practice and behavior.

Simultaneous determination of cephaeline and emetine in ipecac and its preparations using RP-HPLC / 中草药·英文版

Objective: To control the quality of ipecac and its preparations, and to investigate the simultaneous quantitative determination of cephaeline and emetine. Methods: After ultrasonic extraction with acidic methanol solution or direct diluting preparations, cephaeline hydrochloride and emetine hydrochloride in ipecac and its preparations were separated within 20 min using a mixture of acetonitrile-methanol-0.1% phosphoric acid (9:3:88) as the mobile phase on a C18 column by HPLC. UV detector was set at 205 nm. The flow rate was set at 1.0 mL/min. Results: The methodological study showed that a good linear correlation existed in the range of 0.014 560.2184 μg (r = 0.999 97) for cephaeline hydrochloride and 0.0321-0.321 μg (r = 0.999 97) for emetine hydrochloride, respectively. The average recovery of cephaeline hydrochloride and emetine hydrochloride was 96.93% and 99.47%, and the RSD values (n = 9) were 1.31% and 2.02%, respectively. Conclusion: The assay is sensitive, accurate, specific, and applicable to comprehensive evaluation on the quality of ipecac and its preparations. © 2013 Tianjin Press of Chinese Herbal Medicines.

Cyclotide discovery in Gentianales revisited--identification and characterization of cyclic cystine-knot peptides and their phylogenetic distribution in Rubiaceae plants.

Cyclotides are a unique class of ribosomally synthesized cysteine-rich miniproteins characterized by a head-to-tail cyclized backbone and three conserved disulfide-bonds in a knotted arrangement. Originally they were discovered in the coffee-family plant Oldenlandia affinis (Rubiaceae) and have since been identified in several species of the violet, cucurbit, pea, potato, and grass families. However, the identification of novel cyclotide-containing plant species still is a major challenge due to the lack of a rapid and accurate analytical workflow in particular for large sampling numbers. As a consequence, their phylogeny in the plant kingdom remains unclear. To gain further insight into the distribution and evolution of plant cyclotides, we analyzed ∼300 species of >40 different families, with special emphasis on plants from the order Gentianales. For this purpose, we have developed a refined screening methodology combining chemical analysis of plant extracts and bioinformatic analysis of transcript databases. Using mass spectrometry and transcriptome-mining, we identified nine novel cyclotide-containing species and their related cyclotide precursor genes in the tribe Palicoureeae. The characterization of novel peptide sequences underlines the high variability and plasticity of the cyclotide framework, and a comparison of novel precursor proteins from Carapichea ipecacuanha illustrated their typical cyclotide gene architectures. Phylogenetic analysis of their distribution within the Psychotria alliance revealed cyclotides to be restricted to Palicourea, Margaritopsis, Notopleura, Carapichea, Chassalia, and Geophila. In line with previous reports, our findings confirm cyclotides to be one of the largest peptide families within the plant kingdom and suggest that their total number may exceed tens of thousands.

Poaia [Psychotria ipecacuanha (Brot. ) Stoves]: aspectos da memória cultural dos poaieiros de Cáceres - Mato Grosso, Brasil / Ipecac [Psychotria ipecacuanha (Brot. ) Stoves]: aspects of cultural memory of "poaieiros" in Cáceres - Mato Grosso, Brazil

O Brasil está entre os principais exportadores de poaia [Psychotria ipecacuanha (Brot.) Stoves] seguido do Panamá e Costa Rica. A poaia brasileira apresenta alto valor farmacológico das raízes devido aos teores de emetina e cefalina. Este trabalho teve como objetivo descrever como as famílias de poaieiros mantém a memória cultural sobre a Psychotria ipecacuanha (Brot.) Stoves. As informações foram coletadas no município de Cáceres, Mato Grosso, através de entrevista estruturada e observação participante com 20 homens e 10 mulheres, de faixa etária de 45 a 86 anos. Foram citadas as formas de utilização na alimentação para animais, inseticida, carrapaticida, emético, contra diarréias, para alívio de dor de cabeça, contra malária, bronquite e dor no estômago. A raiz é a parte mais usada e a forma de preparo é tintura ou misturada ao fumo, ao vinho ou à cachaça. Poucos entrevistados passaram aos filhos o conhecimento sobre a P. ipecacuanha. A memória cultural sobre a P. ipecacuanha deve-se a vivência, extração e comercialização da planta, e por ouvir as conversas dos pais com amigos. A perda de conhecimento associado a poaia é causada pelo êxodo rural, destruição do habitat com o desmatamento e ocupação agrícola. A extinção da espécie na região contribui para a erosão cultural.

Brazil is among the leading exporters of ipecac [Psychotria ipecacuanha (Brot.) Stoves], followed by Panama and Costa Rica. The roots of Brazilian ipecac have high pharmacological value due to their levels of emetine and cephalin. This study aimed to describe how families of "poaieiros" maintain the cultural memory of Psychotria ipecacuanha (Brot.) Stoves. Information was collected in the city of Cáceres, Mato Grosso State, Brazil, through structured interviews and participating observation involving 20 men and 10 women aged from 45 to 86 years. The cited forms of use were in animal nutrition, as insecticide, acaricide, emetic, against diarrhea, relief from headache, and against malaria, bronchitis and stomach ache. The root is the most used part and its preparation form is tincture or mixed with tobacco, wine or sugarcane. Few interviewees transmitted to their children the knowledge of P. ipecacuanha. The cultural memory of P. ipecacuanha is due to experience, extraction and marketing of the plant, as well as to hearing conversations between parents and friends. The loss of knowledge associated with ipecac is caused by rural exodus, destruction of its habitat due to deforestation and agricultural occupation. The extinction of this species in the region contributes to cultural erosion.

Reproductive studies in ipecac (Psychotria ipecacuanha (Brot. ) stockes; Rubiaceae): pollen development and morphology

The aim of this work was to carry out the reproductive studies on Brazilian accessions of ipecac, Psychotria ipecacuanha. It presented heterostyly, with brevistylous and longistylous flowers. The pollen development was observed from the sections of the anthers embedded in resin. Anther development was normal as usually observed in dicotyledones, displaying four layers: outer epidermis, endothecium, middle layer and inner tapetum. The pollen was bicellular and filled with starch at the microspore stage. Pollen morphology was studied using SEM, which showed pollen polymorphism within and between the two floral morphs. Five types of pollen with reticulate or perforate exine were identified. The characteristics showed that the sexual process was as important as the vegetative propagation for the reproduction of this species.

Foram realizados estudos reprodutivos em acessos brasileiros de poaia, Psychotria ipecacuanha. Poaia apresenta heterostilia, com flores brevistilas e longistilas. O desenvolvimento do pólen foi estudado em cortes de anteras embebidas em resina. O desenvolvimento da antera seguiu o padrão normal para as dicotiledôneas, a qual apresentou quatro camadas: epiderme, endotécio, camada média, e tapete, a mais interna. O pólen apresentou-se bicelular e preenchido com amido no estágio de micrósporo. A morfologia do pólen foi estudada utilizando-se MEV. Foi observado polimorfismo polínico dentro e entre as duas formas florais. Foram identificados cinco tipos de grãos de pólen, com exina reticulada ou perfurada. Em seu hábitat natural, sabe-se que essa espécie apresenta propagação por multiplicação vegetativa, mas as características estudadas demonstraram que o mecanismo sexuado é tão importante para a reprodução dessa espécie quanto à propagação vegetativa.

Reproductive studies on ipecac (Cephaelis ipecacuanha (Brot.) A. Rich; Rubiaceae): meiotic behavior and pollen viability

Reproductive studies were carried out on Brazilian accessions of ipecac, Cephaelis ipecacuanha. Meiotic behavior was studied using the squashing technique. Irregular chromosome segregation in meiosis I and II, many sets of chromosomes in telophase II, micronuclei, incorrect cytoplasm division, incomplete cytokinesis and anomalous post-meiotic products, mainly polyads, were observed. The mean meiotic index was lower than 72%. Pollen viability was analyzed using Alexander solution, and the percentages ranged between brevistylous and longistylous floral morphs (85.3 to 93.1%), and among different localities (82.5 to 92.6%) analyzed. The size of pollen ranged between viable and sterile, and empty and shrunken sterile. In its natural habitat, this species is known to propagate by vegetative multiplication, but sexual reproduction seems to be as important as the vegetative propagation to this species.

Estudos reprodutivos foram realizados em acessos brasileiros de poaia, Cephaelis ipecacuanha. O comportamento meiótico foi estudado usando a técnica de esmagamento. Foi observada segregação irregular de cromossomos durante meiose I e II, muitos grupos de cromossomos em telófase II, micronúcleos, divisão incorreta do citoplasma, citocinese incompleta e produtos pós-meióticos anômalos, principalmente políades. A média do índice meiótico foi inferior a 72%. A viabilidade polínica foi analisada utilizando-se solução de Alexander e a percentagem de pólen viável variou entre as formas florais, brevistila e longistila (85,3% a 93,1%), e entre as diferentes localidades (82,5% a 92,6%) analisadas. O tamanho do pólen variou entre viáveis e inviáveis, e entre os inviáveis vazios e contraídos. Em seu habitat natural, a poaia apresenta propagação por multiplicação vegetativa, mas a reprodução sexuada parece ser tão importante para essa espécie quanto a propagação vegetativa.