Steroidal Alkaloids Research Articles

Estimation of selection criteria through multi-year assessment of variability parameters, association studies and genetic diversity of Solanum khasianum CB Clarke

Solanum khasianum Clarke belonging to the Solanaceae family contains a steroidal alkaloid, solasodine which has commercial and medicinal importance. Solasodine acts as a starting material for the synthesis of steroid hormones, such as oral contraceptives and cortisone. In the present study, a total of 273 accessions were collected out of which 186 accessions rich in solasodine content (>0.8 %) were selected for the study. These selected lines were planted in bed size of 3.6 × 3.6 m in a randomized complete block design (RCBD) with plant to plant spacing of 60 cm with three replications during kharif 2018, kharif 2019, and kharif 2020 for which pooled data were analyzed. Analysis of variance revealed significant differences among the genotypes for all the studied characters revealing high variability among them. Highest GCV (Genotypic coefficient of variation), PCV (Phenotypic coefficient of variation) along with high heritability with high genetic advance was depicted in 100 fresh fruit weight and the number of fruits/plant indicating high variability due to additive genes action which makes it useful in the selection program. Low magnitude between GCV and PCV was observed in the trait plant spread indicating minimum environmental influence and high effect of environment was observed in 100 fresh fruit weight. Based on the genotypic and phenotypic correlation, the number of fruits/plant, plant height, plant spread, and fruit diameter was found to be significantly and positively correlated with100 fresh fruit weight. However, the solasodine content was negatively correlated with 100 fresh fruit weight and the number of fruits/plant. The path coefficient analysis suggested that the number of fruit per plant and plant spread were the direct contributing traits for the fruit yield. In contrast, days to maturity were the direct contributing factor via 100 fresh fruit weight for solasodine content. Diversity using Mahalanobis D2 analysis revealed 80.54% variability which was contributed by the number of fruit per plant, 100 fresh fruit weight, leaf width, leaf length, plant spread, fruit diameter, and days to maturity. The present variability and multivariate analysis among the high solasodine content lines of S. khasianum will help in the breeding, conservation, and varietal development program of this industrially important crop. This is the first report on the variability parameters, association, and diversity study among 186 germplasm of S. khasianum.

Excavating phytochemicals from plants possessing antiviral activities for identifying SARS-CoV hemagglutinin-esterase inhibitors by diligent computational workflow

Coronaviruses (CoVs) belong to a group of RNA viruses that cause diseases in vertebrates including. Newer and deadlier than SARS CoV-2 are sought to appear in future for which the scientific community must be prepared with the strategies for their control. Spike protein (S-protein) of all the CoVs require angiotensin-converting enzyme2 (ACE2), while CoVs also require hemagglutinin-acetylesterase (HE) glycoprotein receptor to simultaneously interact with O-acetylated sialic acids on host cells, both these interactions enable viral particle to enter host cell leading to its infection. Target inhibition of viral S-protein and HE glycoprotein receptor can lead to a development of therapy against the SARS CoV-2. The proposition is to recognize molecules from the bundle of phytochemicals of medicinal plants known to possess antiviral potentials as a lead that could interact and mask the active site of, HE glycoprotein which would ideally bind to O-acetylated sialic acids on human host cells. Such molecules can be addressed as ‘HE glycoprotein blockers’. A library of 110 phytochemicals from Withania somnifera, Asparagus racemosus, Zinziber officinalis, Allium sativum, Curcuma longa and Adhatoda vasica was constructed and was used under present study. In silico analysis was employed with plant-derived phytochemicals. The molecular docking, molecular dynamics simulations over the scale of 1000 ns (1 μs) and ADMET prediction revealed that the Withania somnifera (ashwagandha) and Asparagus racemosus (shatavari) plants possessed various steroidal saponins and alkaloids which could potentially inhibit the COVID-19 virus and even other CoVs targeted HE glycoprotein receptor. Communicated by Ramaswamy H. Sarma

Jerveratrum-Type Steroidal Alkaloids Inhibit β-1,6-Glucan Biosynthesis in Fungal Cell Walls.

ABSTRACTThe limited number of available effective agents necessitates the development of new antifungals. We report that jervine, a jerveratrum-type steroidal alkaloid isolated from Veratrum californicum, has antifungal activity. Phenotypic comparisons of cell wall mutants, K1 killer toxin susceptibility testing, and quantification of cell wall components revealed that β-1,6-glucan biosynthesis was significantly inhibited by jervine. Temperature-sensitive mutants defective in essential genes involved in β-1,6-glucan biosynthesis, including BIG1, KEG1, KRE5, KRE9, and ROT1, were hypersensitive to jervine. In contrast, point mutations in KRE6 or its paralog SKN1 produced jervine resistance, suggesting that jervine targets Kre6 and Skn1. Jervine exhibited broad-spectrum antifungal activity and was effective against human-pathogenic fungi, including Candida parapsilosis and Candida krusei. It was also effective against phytopathogenic fungi, including Botrytis cinerea and Puccinia recondita. Jervine exerted a synergistic effect with fluconazole. Therefore, jervine, a jerveratrum-type steroidal alkaloid used in pharmaceutical products, represents a new class of antifungals active against mycoses and plant-pathogenic fungi.IMPORTANCE Non-Candida albicans Candida species (NCAC) are on the rise as a cause of mycosis. Many antifungal drugs are less effective against NCAC, limiting the available therapeutic agents. Here, we report that jervine, a jerveratrum-type steroidal alkaloid, is effective against NCAC and phytopathogenic fungi. Jervine acts on Kre6 and Skn1, which are involved in β-1,6-glucan biosynthesis. The skeleton of jerveratrum-type steroidal alkaloids has been well studied, and more recently, their anticancer properties have been investigated. Therefore, jerveratrum-type alkaloids could potentially be applied as treatments for fungal infections and cancer.

Anti-inflammatory and Antioxidant Effect of Lycoperoside H against the 1,2-Dimethyl Hydrazine (DMH) Induced Colorectal Cancer in Rats.

In many developed countries, colorectal cancer is a leading cause of morbidity and mortality and its etiology is familiar to be a grouping of nutritional and environmental factors, less physical activity and hereditary factors. Lycoperoside H (LH) is a steroidal alkaloid saponin commonly found in the tomato and exhibited the various pharmacological effects. The aim of the current study was to scrutinized the anticancer effect of LH against 1,2‑Dimethyl Hydrazine (DMH) induced colorectal cancer (CRC) in rats. Subcutaneous injection of DMH (20 mg/kg) was used for induction the CRC and rats were received the oral administration of LH (10, 20 and 40 mg/kg) for 16 weeks. At the end of the investigation, the tumor incidence, weight, and body weight were calculated. Antioxidant enzymes (phase I and II), inflammatory cytokines, lipids and inflammatory markers were all examined. DMH induced rats exhibited the increased tumor incidence, reduced body weight and LH treatment significantly (p < 0.001) suppressed the tumor incidence, and enhanced the body weight. LH treatment significantly (p < 0.001) boosted the level of SOD, GPx, GSH, CAT and suppressed the MDA level. LH treatment suppressed the level cytochrome b5 (Cyto b5), cytochrome P450 (Cyto P450) and boosted the level of glutathione S‑transferase (GST), uridine diphosphoglucuronyltransferase (UDP‑GT) in the liver and colon tissue. LH also decreased the level of cytokines includes interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α); inflammatory mediators like Inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), prostaglandin (PGE2) and nuclear factor kappa B (NF-κB) in the hepatic and colon tissue. We can conclude that LH revealed the anticancer effect against the DMH induced CRC via suppression of inflammation and oxidative stress.

Brief Review on Shatavari (Asparagus racemosus) and Its Medicinal Uses

Asparagus racemosus is another name for shatavari. It belongs to the family asparagaceae. It is an adaptable plant as well. The adaptation of human body to mental and physical stress is said to be achieved by the help of adaptogenic herbs like these.&#x0D; Shatavari is an ayurvedic medicine staple that is made available in the form of analeptic for the wellbeing and vitality of person.&#x0D; It is popular among all therapeutic plants due to various active biochemicals present in it such as steroidal glycosides, saponins (Shatavarins I, II, III, and IV), flavonoids, polyphenols, vitamins and alkaloids (racemosol). Shatavari extracts from its different parts like roots, flowers, leaves and stems are effective in treating female organs which are involved in reproduction.&#x0D; For centuries, ayurvedic medicine has employed shatavari. However, there haven't been enough human trials to recommend it for any medical problem. It is, however, safe to consume it in modest doses, and doing so will allow you to benefit from its antioxidant and immune-boosting properties.&#x0D; Except for being an indigenous medicinal plant which known to be important for its sapogenin content which is the precursor of many active pharmacological steroids, it also has many medicinal uses like antioxytoxic (shatavarin four),spasmodic to uterus, hypertensive as well as hypoglycemic activity, anticoagulant activity, antiviral activity, anticancer and antidysentric activity. In ayurvedic medicine asparagus racemosus is considered as tonic specific to females .Despite of being a herb which is rejuvenating it is used in infertility cases of females, increases libido , enhances ovulation as well as folliculogenesis, also increases lactation.

Extraction and Purification of Secondary Metabolites of Paris polyphylla (Smith) Using Column Chromatography.

Paris polyphylla Smith is an erect and herbaceous plant. It has rich chemical constituents such steroidal saponins, phytosterols, flavonoids, and alkaloids that possess antimicrobial and anticancer activities. In the current investigation, we examined the effect of different solvents in the extraction yield and further purification via column chromatography. The ground powdered rhizomes of P. polyphylla was extracted with 100% hexane, 100% ethyl acetate, 70% ethanol, and 70% methanol. The extracts were filtered, evaporated using a rotary evaporator, concentrated, and measured. Subsequently, the solvent with high extraction yield was further purified using column chromatography. Ethanol produced the highest extraction yield as compared to the other solvents. About 30 fractions were eluted which was pooled into four fractions based on the Rf values and bands observed through thin layer chromatography (TLC) analysis.

Steroidal alkaloids defence metabolism and plant growth are modulated by the joint action of gibberellin and jasmonate signalling.

Steroidal glycoalkaloids (SGAs) are protective metabolites constitutively produced by Solanaceae species. Genes and enzymes generating the vast structural diversity of SGAs have been largely identified. Yet, mechanisms of hormone pathways coordinating defence (jasmonate; JA) and growth (gibberellin; GA) controlling SGAs metabolism remain unclear. We used tomato to decipher the hormonal regulation of SGAs metabolism during growth vs defence tradeoff. This was performed by genetic and biochemical characterisation of different JA and GA pathways components, coupled with in vitro experiments to elucidate the crosstalk between these hormone pathways mediating SGAs metabolism. We discovered that reduced active JA results in decreased SGA production, while low levels of GA or its receptor led to elevated SGA accumulation. We showed that MYC1 and MYC2 transcription factors mediate the JA/GA crosstalk by transcriptional activation of SGA biosynthesis and GA catabolism genes. Furthermore, MYC1 and MYC2 transcriptionally regulate the GA signalling suppressor DELLA that by itself interferes in JA-mediated SGA control by modulating MYC activity through protein-protein interaction. Chemical and fungal pathogen treatments reinforced the concept of JA/GA crosstalk during SGA metabolism. These findings revealed the mechanism of JA/GA interplay in SGA biosynthesis to balance the cost of chemical defence with growth.

Two new steroidal alkaloids with cytotoxic activities from the roots of Veratrum grandiflorum Loes.

• Two new steroidal alkaloids were obtained from roots of Veratrum grandiflorum Loes. • Their structures were elucidated on the basis of extensive spectroscopic techniques. • Compounds 3 , 4 exhibited moderate cytotoxic effects against four cancer cell lines. Two new steroidal alkaloids, Grandivine A and B ( 1 and 3 ), together with eight known related compounds were obtained from the roots of Veratrum grandiflorum Loes. The structures of the compounds were elucidated though extensive spectroscopic techniques. Compounds 3 and 4 exhibited moderate cytotoxic effects against four cancer cell lines.

Deciphering the Synergistic Mechanism of Cortistatin towards Cancer Targets Using Network Pharmacology Approach

Abstract: Steroidal alkaloid cortistatin is a promising marine natural compound isolated from marine sponges Corticium simplex. Experimental studies and clinical evidence have shown that cortistatin and its derivatives have a curative effect in patients with autoimmune disorders, HIV infection and several types of cancer. The objective of our study was to examine the potential cancer-related therapeutic objectives of Cortistatin using a network pharmacology method, which is a computational approach, including inverse pharmacophore research, enrichment analysis, molecular docking and dynamics study. Systematic protocol also involves the assessment of ADMET parameters to define the pharmacokinetic profile of cortistatin. Inverse pharmacophore search method was used for computational target fishing and target proteins were ranked based on the graph theory approach. Cancer target proteins, namely HSP90, EGFR, CDK2, MMP13, MAPK13, AR, ESR1, PTPN11 and SRC, were classified as top-ranking proteins according to graph theory parameters, namely MCC, DMNC, MNC, Degree(Local-based methods), EPC, Bottleneck, Eccentricity, Closeness, Radiality, Betweenness, Stress(Global-based methods) and Clustering Coefficient. Enrichment assessment established on Gene Ontology and pathway analysis of these proteins that play a vital role in cancer pathways, FaxO Signalling pathways, Ras Signalling pathways and tyrosine metabolism. Molecular docking and dynamic simulation studies of cortistatin with proposed target proteins were found to be stable and conformers generated after 3ns were consistent with stable inter-molecular interactions. The current study summarized here would provide a broad perspective on the therapeutic potential of cortistatin and provide new insights into the future development of cancer therapy strategies. Key words: Cortistatin, Marine natural product, Molecular docking, Network pharmacology, Cancer targets, Pharmacokinetics.

Tomatidine reduces Chikungunya virus progeny release by controlling viral protein expression.

Tomatidine, a natural steroidal alkaloid from unripe green tomatoes has been shown to exhibit many health benefits. We recently provided in vitro evidence that tomatidine reduces the infectivity of Dengue virus (DENV) and Chikungunya virus (CHIKV), two medically important arthropod-borne human infections for which no treatment options are available. We observed a potent antiviral effect with EC50 values of 0.82 μM for DENV-2 and 1.3 μM for CHIKV-LR. In this study, we investigated how tomatidine controls CHIKV infectivity. Using mass spectrometry, we identified that tomatidine induces the expression of p62, CD98, metallothionein and thioredoxin-related transmembrane protein 2 in Huh7 cells. The hits p62 and CD98 were validated, yet subsequent analysis revealed that they are not responsible for the observed antiviral effect. In parallel, we sought to identify at which step of the virus replication cycle tomatidine controls virus infectivity. A strong antiviral effect was seen when in vitro transcribed CHIKV RNA was transfected into Huh7 cells treated with tomatidine, thereby excluding a role for tomatidine during CHIKV cell entry. Subsequent determination of the number of intracellular viral RNA copies and viral protein expression levels during natural infection revealed that tomatidine reduces the RNA copy number and viral protein expression levels in infected cells. Once cells are infected, tomatidine is not able to interfere with active RNA replication yet it can reduce viral protein expression. Collectively, the results delineate that tomatidine controls viral protein expression to exert its antiviral activity. Lastly, sequential passaging of CHIKV in presence of tomatidine did not lead to viral resistance. Collectively, these results further emphasize the potential of tomatidine as an antiviral treatment towards CHIKV infection.

Genotype-dependent and temperature-induced modulation of secondary metabolites, antioxidative defense and gene expression profile in Solanum viarum Dunal

The medicinal plant, Solanum viarum Dunal includes a number of compounds with important pharmacological effects. The effect of temperature (heat and cold) stress on growth, secondary metabolite levels and defense mechanism in two genotypes (prickled and prickleless) of S. viarum was evaluated. The two thermal regimes (4 °C for cold and 35 °C for heat stress) were found to be the most optimum for the accumulation of metabolites (steroidal alkaloids and glycoalkaloids, phenolic acids as well as flavonoids). Prickleless genotype showed 4.08 and 1.51 fold higher total alkaloids/glycoalkaloid content at 35 °C and 4 °C temperatures respectively, in comparison with prickled plants. Prickleless plants also registered 2.03 and 2.28 higher fresh and dry biomass accumulation respectively, under cold stress over the control plants. The accretion of phenolic and flavonoid compounds under heat and cold stress is mainly contributed by gallic and ferulic acid in both the genotypes. The quantitative real-time PCR expression analysis showed the abundance of gene transcript involve in the biosynthesis of alkaloids/glycoalkaloid and phenolics/flavonoids that corroborates with the accumulation of their respective metabolites in prickleless plants under temperature stress. Results revealed that the prickleless plants inhibiting the reactive oxygen species (ROS) mediated oxidative damage by activating the enzymatic (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase) and non-enzymatic (alkaloids, phenols, flavonoids, carotenoids and proline) antioxidants mechanism that confirms its efficiency to tolerate thermal stress under both the thermal regimes. The overall result showed that the prickleless genotype served as a better accumulator of biomass, secondary metabolites with an improved antioxidative mechanism in comparison to prickled genotype. Our findings suggest that thermal stress responses differ significantly between genotypes, emphasizing the necessity of maintaining genotypic diversity in adaptive evolution as in the context of global warming and climate change. The present study also opens the possibility of exploiting improved prickleless genotype of S. viarum as a potential system that can be cultivated under different temperature stress conditions to attain the phytopharmaceutical benefits.

Chemical profiling of Fritillariae thunbergii Miq prepared by different processing methods reveals two new quality markers: Zhebeininoside and imperialine-3-β-D-glucoside

Fritillariae thunbergii Miq (FTM)exhibit versatile biological activities including the significant antitussive and expectorant activities. As a herbal medicine, the therapeutic effects of FTM may be expressed by multi-components which have complicated integration effects on multi-targets. With the time going, the different processing methods of FTM has been changed a lot. Thus，the study described the effect of processing methods to FTM and its quality. Studies were undertaken by using UHPLC-LTQ Orbitrap MS and pharmacodynamic models. All reagents were involved of analytical grade. While a HPLC-ELSD's method has been developed and validated, a certified Quality System is conformed to ICH requirements. The experimental animals followed the animal welfare guidelines. We aimed to found the differences after the different processing methods of FTM, and to demonstrate the changes could be selected as quality control indicators, and established a method for simultaneous determination of these for quality control. we have previously found two new steroidal alkaloids: zhebeininoside and imperialine-3-β-D-glucoside from the different processing methods of FTM, which is the difference between the different processing methods of FTM, mainly on the steroidal alkaloids. The activity analysis of zhebeininoside, imperialine-3-β-D-glucoside, verticine and verticinone showed that the mouse model of cough expectorant has antitussive effect. The positive drug selected was dextromethorphan syrup. The positive group showed biological activity, but the blank group showed nothing. The model group showed illness which means that the model was effective. There are two ways of the mechanism of action of the expectorant action which can make sputum thin, reduce its viscosity, and be easy to cough up, or can accelerate the movement of mucous cilia in the respiratory tract and promote the discharge of sputum. In our study, the content of phenol red was significantly reduced in the administration group. To sum up, our results suggest that zhebeininoside and other three components cloud be selected as quality control indicators, and a method for simultaneous determination of zhebeininoside and other three components was established for quality control.

Aqueous extract of Solanum nigrum attenuates Angiotensin-II induced cardiac hypertrophy and improves cardiac function by repressing protein kinase C-ζ to restore HSF2 deSUMOlyation and Mel-18-IGF-IIR signaling suppression

Ethnopharmacological relevanceSolanum nigrum, commonly known as Makoi or black shade has been traditionally used in Asian countries and other regions of world to treat liver disorders, diarrhoea, inflammatory conditions, chronic skin ailments (psoriasis and ringworm), fever, hydrophobia, painful periods, eye diseases, etc. It has been observed that S. nigrum contains substances, like steroidal saponins, total alkaloid, steroid alkaloid, and glycoprotein, which show anti-tumor activity. However; there is no scientific evidence of the efficacy of S. nigrum in the treatment of cardiac hypertrophy. AimTo investigate the ability of S. nigrum to attenuate Angiotensin II - induced cardiac hypertrophy and improve cardiac function through the suppression of protein kinase PKC-ζ and Mel-18-IGF-IIR signaling leading to the restoration of HSF2 desumolyation. Materials and methodsCardiomyoblast cells (H9c2) were challenged with 100 nM Angiotensin-II (AngII) for 24 h and were then treated with different concentration of S.nigrum or Calphostin C for 24 h. The hypertrophic effect in cardiomyoblast cells were determined by immunofluorescence staining and the modulations in hypertrophic protein marker along with Protein Kinase C-ζ, MEL18, HSF2, and Insulin like growth factor II (IGFIIR), markers were analyzed by western blotting. In vivo experiments were performed using 12 week old male Wistar Kyoto rats (WKY) and Spontaneously hypertensive rats (SHR) separated into five groups. [1]Control WKY, [2] WKY –100 mg/kg of S.nigrum treatment, [3] SHR, [4] SHR-100 mg/kg of S.nigrum treatment, [5] SHR-300 mg/kg of S.nigrum treatment. S. nigrum was administered intraperitoneally for 8 week time interval. ResultsWestern blotting results indicate that S. nigrum significantly attenuates AngII induced cardiac hypertrophy. Furthermore, actin staining confirmed the ability of S. nigrum to ameliorate AngII induced cardiac hypertrophy. Moreover, S. nigrum administration suppressed the hypertrophic signaling mediators like Protein Kinase C-ζ, Mel-18, and IGFIIR in a dose-dependent manner and HSF2 activation (restore deSUMOlyation) that leads to downregulation of IGF-IIR expression. Additionally in vivo experiments demonstrate the reduced heart sizes of S. nigrum treated SHRs rats when compared to control WKY rats. ConclusionCollectively, the data reveals the cardioprotective effect of S. nigrum inhibiting PKC-ζ with alleviated IGF IIR level in the heart that profoundly remits cardiac hypertrophy for hypertension-induced heart failure.

Modulation of Secondary Metabolites: A Halotolerance Strategy of Plant Growth Promoting Rhizobacteria Against Sodium Chloride Stress.

An experiment was conducted to evaluate the role of bacterial secondary metabolites against induced salt stress. Five bacterial strains were isolated from three different habitats: Khewra salt range, oily sludge field in Chakwal, and garden soil of Quaid-i-Azam University Islamabad, Pakistan. The 16S rRNA gene and BLAST analysis of bacterial strains showed 99% sequence similarity with Pseudomonas putida AMUPP-2 (KM435273), Lysinibacillus sphaericus OUG29GKBB (KM972671), Bacillus pumilus MB431 (KP723538) isolated from salt range, Pseudomonas fluorescens B8 (KF010368) from garden soil and Exiguobacterium aurantiacum SPD2 (KX121703) from oily sludge, respectively. Pseudomonas fluorescens produced 294.98 µg/g of proline in the M9 medium supplemented with 125 mM NaCl, but its growth rate was decreased from 1.81 to 0.37. The P. putida showed faster growth rate even than control at 125 mM NaCl. B. pumilus and L. sphaericus did not show any decline in growth rate up to 100 mM NaCl. The synthesis of new amino acids were recorded at 125 mM NaCl stress, e.g., Pro, Leu, Arg in P. fluorescens and L. sphaericus, Pro, Lys, Phe, Ala in P. putida, Lys, Ala in B. pumilus, Met, Val, and Ala in E. aurantiacum. Liquid chromatography-mass spectrometry analysis of ethyl acetate extract of P. putida and L. sphaericus demonstrated that NaCl (125mM) induced the production of 3-oxo-C12 homoserine lactone, oxosteroids, and steroid esters in addition to steroidal alkaloid lysophosphatidylcholines, antibiotics phenazine-1 carboxamide, 2,4-diacetyl phloroglucinol, carbazole, phosphatidylcholine, phosphatidyl ethanol amine, and salicylic acid as signaling compound. It was concluded that P. putida and L. sphaericus could be exploited for the production of secondary metabolites that have a wide range of implications in biotic and abiotic stresses and for the production of important pharmaceutical products.

3-Epipachysamine B suppresses proliferation and induces apoptosis of breast cancer cell via PI3K/AKT/mTOR signaling pathway

3-Epipachysamine B is a natural steroidal alkaloid isolated from Pachysandra terminalis Sieb. et Zucc. (known locally as Kunxianqi). Kunxianqi contains numerous compounds with demonstrated activity against breast cancer (BRCA). However, it is unknown whether 3-epipachysamine B also has anti-BRCA efficacy. In the present study, we employed network pharmacology technology to search and find potential molecular targets of 3-epipachysamine B. We applied cell proliferation, apoptosis, and western blotting assays to test the predicted key targets and the effects of 3-epipachysamine B against BRCA. Network pharmacology disclosed 80 potential BRCA-related targets of 3-epipachysamine B and assigned them to 75 signaling pathways. Of these, the most highly enriched was the PI3K/AKT signaling pathway. PIK3R1, AKT1, and mTOR had high degrees and betweenness centrality in protein-protein interaction network and are associated with PI3K/AKT signaling. Molecular docking and molecular dynamics simulation indicated strong binding between 3-epipachysamine B and PIK3R1, AKT1, and mTOR. 3-Epipachysamine B repressed the proliferation and induced the apoptosis of BRCA cells, as well as downregulated P-AKT/AKT, P-mTOR/mTOR, and P-PI3K/PI3K in the cells. The PI3K inhibitor LY294002 augmented these changes. Hence, 3-epipachysamine could also prove effective as an anticancer agent in future animal tumor model and human clinical breast cancer trials. Successful validation results could lead to a safe and effective new breast cancer treatment that improves patient prognosis and quality of life.

Potential of Steroidal Alkaloids in Cancer: Perspective Insight Into Structure-Activity Relationships.

Steroidal alkaloids contain both steroidal and alkaloid properties in terms of chemical properties and pharmacological activities. Due to outstanding biological activities such as alkaloids and similar pharmacological effects to other steroids, steroidal alkaloids have received special attention in anticancer activity recently. Substituted groups in chemical structure play markedly important roles in biological activities. Therefore, the effective way to obtain lead compounds quickly is structural modification, which is guided by structure–activity relationships (SARs). This review presents the SAR of steroidal alkaloids and anticancer, including pregnane alkaloids, cyclopregnane alkaloids, cholestane alkaloids, C-nor-D-homosteroidal alkaloids, and bis-steroidal pyrazine. A summary of SAR can powerfully help to design and synthesize more lead compounds.

Anthelmintic potency of Rumex crispus L. extracts against Caenorhabditis elegans and non-targeted identification of the bioactive compounds

Traditional healers and ethnoveterinary therapists use several medicinal plants, such as Rumex crispus L., to treat endoparasite infections. R. crispus has been established by researchers to be effective agasint a few parasitic worms. In this study, we evaluated the potency of R. crispus extracts on the model organism, Caenorhabditis elegans and the bioactive compounds of the extracts were also identified. The solvent extracts of R. crispus were tested against C. elegans for up to 72 h. The effect of the extracts on C. elegans was examined using light microscopy (LM) and scanning electron microscopy (SEM). LM and SEM analysis showed damage on the body wall, reduced body and slight modifications of the nematode organs. The lethality test reveals a significant reduction in the viability of the nematode with the water extract of leaf (LF-WAE), among others, having the strongest potency against the nematode, with 83% lethality. Anlysis done with Fourier-transform infrared spectroscopy (FTIR) spectra reveals various characteristic vibration bands and fingerprint bands at 3400–600 cm−1, identifying phenols, organic acids, aromatics, amines, among others in the plant. The compounds were identified with liquid chromatography-mass spectrometry (LC-MS), under the categories of flavonoids, steroidal alkaloids and proanthocyanidin. In conclusion, this study confirmed that R. crispus has anthelmintic potential, using standardised C. elegans models as a tool and suggests that there could be novel compounds yet to be explored in the studied plant that could be of great benefit to livestock and humans.

Steroidal Alkaloids with a Potent Analgesic Effect Based on N-type Calcium Channel Inhibition.

Two distinctive alkaloids with 6/6/6/5/6/6 fused rings, in which a previously unidentified linkage of C-12/23 generates a rigid skeleton, resulting in a new subtype of steroidal alkaloid, were isolated from Veratrum grandiflorum. Compounds 1 and 2 showed potent analgesic effects in vivo, superior to the well-known analgesic, pethidine (Dolantin), likely by inhibiting CaV2.2 voltage-gated calcium channels.

Tomatidine Suppresses the Destructive Behaviors of Fibroblast-Like Synoviocytes and Ameliorates Type II Collagen-Induced Arthritis in Rats.

Fibroblast-like synoviocytes (FLSs) are the prominent non-immune cells in synovium and play a pivotal role in rheumatoid arthritis (RA) pathogenesis. Searching for natural compounds that may suppress the pathological phenotypes of FLSs is important for the development of RA treatment. Tomatidine (Td), a steroidal alkaloid derived from the solanaceae family, has been reported to have anti-inflammatory, anti-tumor and immunomodulatory effects. However, its effect on RA remains unknown. Here, we examined the inhibitory effect of Td on TNFα-induced arthritic FLSs, and subsequently investigated its therapeutic effect on collagen-induced arthritis (CIA) rats. Our results revealed that Td significantly inhibited TNFα-induced proliferation and migration of arthritic FLSs. In addition, we found that Td treatment could efficaciously ameliorate synovial inflammation and joint destruction of rats with CIA. Both in vitro and in vivo studies showed that Td significantly suppressed the production of pro-inflammatory cytokines including IL-1β, IL-6 and TNFα, and downregulated the expression of MMP-9 and RANKL. Further molecular mechanism studies revealed that the inhibitory effect of Td on RA might attribute to the decreased activations of MAPKs (ERK and JNK) and NF-κB. These findings provide evidence that Td has the potential to be developed into a complementary or alternative agent for RA therapy.

Tomatidine provides mitophagy-independent neuroprotection after ischemic injury.

Cerebral ischemia is one of the leading causes of human mortality and disability worldwide. The treatment of cerebral ischemia is refractory due to its short therapeutic window and lack of effective clinical drugs. Mitophagy, the autophagic elimination of damaged mitochondria, attenuates neuronal injury in cerebral ischemia, indicating the potential of mitophagy inducers as therapies for cerebral ischemia. We previously determined that, by enhancing autophagy flux, the steroidal alkaloid tomatidine can function as a neuroprotective agent against ischemic injury. However, its effects on mitophagy remain unknown. For this purpose, neuroblastoma cell lines Neuro‐2a and SH‐SY5Y were subjected to ischemic injury induced by oxygen–glucose deprivation/reperfusion (OGD/R) and then treated with tomatidine. OGD/R induced a general decrease of cellular contents, and this study revealed that tomatidine had no impact on mitophagy. In addition, tomatidine did not affect mitochondrial contents, including translocase of outer mitochondrial membrane 20 and voltage‐dependent anion channel 1, in either OGD/R‐treated or intact SH‐SY5H cells. Our results indicate that tomatidine exhibits its neuroprotective effects by enhancing autophagy, but in a potentially mitophagy‐independent manner, and provide insights for further investigation into its mechanism(s) and potential therapeutic use against cerebral ischemia.