Semi-purified Fractions Research Articles

Antiproliferative and Pro-apoptotic Activities of Tournefortia mutabilis vent. Leaves on the Human Breast Adenocarcinoma Cell Line (MCF-7)

Background: Breast cancer is the most common cancer among women worldwide, impacting not only the patients but also their families and communities. Tournefortia mutabilis vent. is a plant endemic to Mexico, traditionally used in Zapotec medicine for the treatment of cancer. Objectives: This study aims to evaluate the effects of the chloroformic extract of T. mutabilis vent. leaves on cell proliferation and cell death in MCF-7 cells. Methods: The effect of the extract on MCF-7 cell proliferation was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet staining. Apoptosis was evaluated through fluorescein diacetate/propidium iodide staining and caspase-3, -6, and -9 activity assays. Results: The half-maximal inhibitory concentration (IC50) of the T. mutabilis vent. extract on MCF-7 cell proliferation at 48 hours and 72 hours after treatment was 86.4 µg/mL and 2.74 µg/mL, respectively. We observed that the extract and its semi-purified fractions induced cell death through the activation of caspases 3, -6, and -9. Conclusions: Tournefortia mutabilis vent. is a potential source of compounds with antiproliferative and pro-apoptotic activities on the MCF-7 cell line, primarily through the intrinsic pathways of apoptosis.

Cytotoxic Effect of Clinacanthus nutans Semi-purified Fraction (SF1) in Combination with Cisplatin against Human Cervical Cancer

Cytotoxic Effect of Clinacanthus nutans Semi-purified Fraction (SF1) in Combination with Cisplatin against Human Cervical Cancer

Antifungal effect of a metabolite of Pseudomonas aeruginosa LV strain on azole-resistant Candida albicans

Candida albicans remains the most common agent of candidiasis worldwide. This yeast is generally sensitive to most antifungals, however, the emergence of azole-resistant C. albicans has been reported. In addition, this microorganism can form biofilms on various surfaces, making it difficult to treat infections. In this study, the effect of secondary metabolites of Pseudomonas aeruginosa strain LV on planktonic and sessile cells of C. albicans, with different genotypes and susceptibility profile to fluconazole and voriconazole, was evaluated. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the semi-purified fraction F4a ranged from 1.56 to 6.25 μg/mL and 6.25 to 25 μg/mL, respectively. Fluopsin C appears to be the antifungal component of F4a. The semi-purified fraction and fluopsin C showed fungicidal activity, dose and time dependent. F4a caused severe damage to the morphology and ultrastructure of planktonic fungal cells, and significantly reduced the viability of 24-hour biofilms, with MIC for sessile cells from 12.5 to 25.0 μg/mL. However, cytotoxicity was detected in mammalian cells for F4a and fluopsin C at concentrations that showed antifungal activity. These results indicate that fluopsin C may be a prototype for the development of new antifungals for C. albicans.

Evaluation of Resistance Induction Promoted by Bioactive Compounds of Pseudomonas aeruginosa LV Strain against Asian Soybean Rust.

Pseudomonas are known as higher producers of secondary metabolites with antimicrobial properties and plant growth promoters, including resistance induction. These mechanisms should be an alternative to pesticide use in crop production. Phakopsora pachyrhizi causes Asian soybean rust, representing a high loss of yield around the world. The objective of this paper was to evaluate the application of secondary metabolites produced by Pseudomonas aeruginosa LV strain from the semi-purified fraction F4A in soybean plants to induce plant resistance against P. pachyrhizi in field conditions. The experimental design was performed in randomized blocks with three replicates using two F4A doses (1 and 10 μg mL-1) combined or not with fungicides (Unizeb Gold® or Sphere Max®). The control treatment, with Uni + Sph, saponins, flavonoids, and sphingolipids, showed higher intensities in the plants. In contrast, plants treated with the F4A fraction mainly exhibited fatty acid derivatives and some non-identified compounds with nitrogen. Plants treated with Sphere Max®, with or without F4A10, showed higher intensities of glycosylated flavonoids, such as kaempferol, luteolin, narigenin, and apigenin. Plants treated with F4A showed higher intensities of genistein and fatty acid derivatives. These increases in flavonoid compound biosynthesis and antioxidant properties probably contribute to the protection against reactive oxygen species (ROS).

Phytochemical Analysis and Biological Activities of Solvent Extracts of Two Traditionally Used Medicinal Plants

The plants Buddleja asiatica and Buddleja paniculata are two closely related species of which the former has widespread uses in traditional medicine while the latter remains relatively unexplored. The present study focused on the analysis of phytoconstituents and the estimation of different biological activities of extracts and semi-purified fractions. The ethyl acetate stem fraction and methanolic extract of the leaf of B. asiatica displayed the highest phenolic contents of 390.98 ± 5.32 and 383.72 ± 7.31 mg GAE/g. TFC content was measured as 207.33 ± 0.34 and 138.58 ± 0.53 mg QE/g for ethyl acetate leaf and stem fraction of B. asiatica. The highest antioxidant activity was displayed by crude stem extract of B. paniculata with IC50 35.65±0.61 µg/mL in DPPH assay followed by ethyl acetate stem fraction of B. asiatica with IC50 36.17±0.92 µg/mL. The DCM leaf fraction and stem fraction of B. asiatica were found active against gram-negative and gram-positive bacteria. Both plants displayed mild to no toxicity against the brine shrimp nauplii. Median lethal concentration (LC50) ranged from 346.96 ± 25.39 in hexane fraction of leaf to 2719.32 ± 706.5 µg/mL in crude methanol extract of B. paniculata. The present study showed these medicinal plants are rich in an important class of organic compounds that play a significant role in the cure of bacterial infection which ultimately supports pharmaceutical research.

Anti-Diabetic, Anti-Cholinesterase, and Anti-Inflammatory Potential of Plant Derived Extracts and Column Semi-Purified Fractions of Ficus benghalensis.

The present study aimed to investigate the in-vitro anti-diabetic, anti-cholinesterase, and anti-inflammatory potential of extracts from different parts of Ficus benghalensis, including leaves, stem, and roots, as well as isolated column fractions (F-B-1 C, F-B-2 C, F-B-3 C, and F-B-4 C). The extracts and subsequent fractions were evaluated for their inhibitory activity against key enzymes involved in diabetes [α-glucosidase and α-amylase], neurodegenerative diseases [acetylcholinesterase and butyrylcholinesterase], and inflammation (cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX)). The results showed that F. benghalensis leaf extract exhibited the highest α-glucosidase inhibitory activity (73.84%) and α-amylase inhibitory activity (76.29%) at 1000 µg/mL. The stem extract (65.50%) and F-B-2 C fraction (69.67%) also demonstrated significant α-glucosidase inhibitory activity. In terms of anti-cholinesterase activity, the extracts of roots, leaves, and stem showed promising inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with half maximal inhibitory concentration (IC50) values ranging from 50.50 to 474.83 µg/mL. The derived fractions (F-B-1 C, F-B-2 C, F-B-3 C, and F-B-4 C) also exhibited notable inhibition of AChE and BChE, with IC50 values from 91.85 to 337.94 µg/mL. Moreover, the F-B-3 C fraction demonstrated the highest COX-2 inhibitory potential (85.72%), followed by F-B-1 C (83.13%), the stem extract (80.85%), and the leaves extract (79.00%). The F-B-1 C fraction showed the highest 5-LOX inhibitory activity (87.63%), while the root extract exhibited the lowest inhibition (73.39%). The results demonstrated promising bioactivity, suggesting the potential of F. benghalensis as a source of natural compounds with therapeutic applications. Further studies are required to identify and isolate the active components responsible for these effects and to evaluate their in-vivo efficacy and safety.

Induction of Apoptosis in Candida albicans by Semi-purified Fraction of Hypnea musciformis

Induction of Apoptosis in Candida albicans by Semi-purified Fraction of Hypnea musciformis

Synergistic Antifungal Interaction between Pseudomonas aeruginosa LV Strain Metabolites and Biogenic Silver Nanoparticles against Candida auris.

Candida auris has been found to be a persistent colonizer of human skin and a successful pathogen capable of causing potentially fatal infection, especially in immunocompromised individuals. This fungal species is usually resistant to most antifungal agents and has the ability to form biofilms on different surfaces, representing a significant therapeutic challenge. Herein, the effect of metabolites of Pseudomonas aeruginosa LV strain, alone and combined with biologically synthesized silver nanoparticles (bioAgNP), was evaluated in planktonic and sessile (biofilm) cells of C. auris. First, the minimal inhibitory and fungicidal concentration values of 3.12 and 6.25 μg/mL, respectively, were determined for F4a, a semi-purified bacterial fraction. Fluopsin C and indolin-3-one seem to be the active components of F4a. Like the semi-purified fraction, they showed a time- and dose-dependent fungicidal activity. F4a and bioAgNP caused severe changes in the morphology and ultrastructure of fungal cells. F4a and indolin-3-one combined with bioAgNP exhibited synergistic fungicidal activity against planktonic cells. F4a, alone or combined with bioAgNP, also caused a significant decrease in the number of viable cells within the biofilms. No cytotoxicity to mammalian cells was detected for bacterial metabolites combined with bioAgNP at synergistic concentrations that presented antifungal activity. These results indicate the potential of F4a combined with bioAgNP as a new strategy for controlling C. auris infections.

Anticancer and antioxidant activities of ethanolic extract and semi-purified fractions from guava and mango seeds

Mango (Mangifera indica) and guava (Psidium guajava) seeds have several pharmaceutical applications and biological activities because as they have been recognized with different bioactive molecules (phenolic compounds) such as flavonoids, phenolic acids, and catechins, so they have antioxidant and anticancer activities. The aim of the present study was to assess in vitro antioxidant and anticancer activities of successive extracts and semi-purified fractions from mango seeds. In this work, mango and guava seeds were collected and extracted using two solvents (ethanol 70% and ethyl acetate) followed by phytochemical screening and determination of biological activities such as antioxidant activity using five assays (DPPH, ABTS, KMnO4, Methylene blue and H2O2) additionally the antiradical activity and hybrid reaction for ethanolic extract of mango seeds as promising extract. The total phenolic, flavonoid, and catechin compounds were determined for all successive extracts, and finally, the anticancer activity of extracts was evaluated using MTT assay against HepG2 cell line and phenolic compounds were identified by HPLC. The phytochemical screening and TLC showed the primary investigation for phenolic compounds of ethanol extracts of both kind of seeds and only ethyl acetate of guava extract as promising extracts. However, HPLC determination of these three extracts showed high amount of gallic acid, naringenin, ellagic acid, and ferulic acid as they have anticancer and antioxidant activities. The antioxidant tests showed that the ethanolic mango extract is the highest antioxidant extract against DPPH by 84.0%, but recorded 82.0% with methylene blue and ABTS assays when compared with ascorbic acid. The ethyl acetate of guava extract showed strong cytotoxic effect with IC50 75.5 μg/mL against HepG2 cell line in all tested concentrations. From the obtained results, it could be concluded that mango ethanolic extract and its fractions are the most promising as antioxidants and ethyl acetate of guava extract the most promising in the anticancer activity.

Growth Inhibitory Activities and Feeding Deterrence of Solanaceae-Based Derivatives on Fall Armyworm

Spodoptera frugiperda is a pest of worldwide importance, responsible for significant economic losses, mainly in maize crops. The use of botanical derivatives emerges as a promising alternative to control this insect pest. In this work, we evaluated the effect of ethanolic extracts (EE) and semi-purified fractions of Acnistus arborescens and Datura stramonium (Solanaceae) on the biological development of S. frugiperda and the effects of the semi-purified fractions on feeding behavior of 4th instar caterpillars. Crude extracts and fractions caused lethal and sublethal effects, namely increasing both duration of larval and pupal stages as well as deformities in adults, and decreasing weight of pupae. In turn, the effects on feeding deterrence were more pronounced in treatments with A. arborescens fractions. Our results highlight the potential of EE from solanaceous species as a source of allelochemicals that can be used in the integrated management of S. frugiperda.

Chemical Constituents from Macaranga lowii King ex Hook.f. (Euphorbiaceae) and Their Effects on Antioxidant Activity

The genus Macaranga comprises about 300 species worldwide and is commonly distributed in the tropical region of Africa, Southeast Asia, China and India. The plant of this genus was reported to possess various medicinal properties which have been traditionally used to treat various illnesses such as fungal infection, malaria, stomachache, and diarrhoea. This genus also contains many phenolic compounds, particularly prenylated flavonoids and stilbenoids, followed by terpenoids and tannins. The phytochemical study on the crude acetone extract from the stem bark of Macaranga lowii was carried out to isolate chemical constituents using several chromatographic techniques and characterise the structure of the isolated compounds using various spectroscopic methods. The powdered stem bark of M. lowii (2.05 kg) was macerated in acetone for 24 hours at room temperature and repeated three times. The crude extract (38.8 g) was dissolved in methanol (200 mL) and fractionated with diethyl ether to reduce tannin and yield a crude extract with less tannin (10.9 g). The crude extract was subjected to vacuum liquid chromatography (VLC) to give nine semi-purified fractions (ML1-9). Fractionation and purification of fraction ML2 (448 mg) yielded compound 1 (6.2 mg) and 3 (0.3 mg). Fraction ML7 (220 mg) was subjected to radial chromatography (RC) to give a pure compound 2 (2.1 mg). The compounds were elucidated using Nuclear Magnetic Resonance (NMR), Ultraviolet-Visible (UV-Vis), Infrared (IR) and Mass spectrometry (MS), as well as comparison with the previous literature. Two phenolic compounds namely pentadecyl ferulate (1) and scopoletin (2) together with a sterol known as stigmasterol (3) were purified successfully from the stem bark of M. lowii. Scopoletin (2) exhibited moderate activity against DPPH radical scavenging with the IC50 value of 34.82 mM.

Cosmeceutical Potentials of Equisetum debile Roxb. ex Vaucher Extracts

Trends in skin and hair treatments focus on natural products due to undesired effects of chemically synthetic ingredients. This study aims to investigate the cosmeceutical effects of Equisetum debile (horsetail) extracts relating to anti-hyperpigmentation via tyrosinase, anti-wrinkle formation via matrix metalloproteinases (MMPs), and anti-androgenic alopecia via 5α-reductase. Ethanolic extracts were sequentially partitioned into semi-purified fractions hexane, dichloromethane, ethyl acetate, methanol, and methanol insoluble residue. The ethyl acetate fraction possessed the highest total phenolic content (39.24 ± 0.72 mg gallic acid/g), the strongest anti-tyrosinase activities (583.33 ± 23.59 mg kojic acid/g), and potent collagenase inhibitions (IC50 MMP-1 and MMP-2 of 0.82 ± 0.09 and 0.94 ± 0.11 mg/mL, respectively). All extracts showed considerable inhibitions of 5α-reductase ranging from 44.59 ± 0.40 to 83.07 ± 3.46% with the strongest activity from the dichloromethane fraction (1.48 ± 0.06 mg finasteride/g). In conclusion, E. debile extracts exhibit cosmeceutical potentials. This study suggests that the E. debile ethyl acetate fraction could be used as a promising ingredient to organically treat hyperpigmentation and delay the skin aging process. In addition, compared to the current recommended intake of finasteride (1 mg/day) for androgenic alopecia, the dichloromethane fraction is proposed as an alternative source to naturally remediate hair loss.

Fluopsin C: A Review of the Antimicrobial Activity against Phytopathogens

Fluopsin C (FlpC) is an organocupric secondary metabolite with low-molecular-weight, produced by some Pseudomonas and Streptomyces bacteria. The compound was identified in 1970 as prismatic dark-green crystals, with strong antimicrobial activity against several human and phytopathogens. Due to its high cytotoxicity, research on this compound decreased after the 1970s. During the early 2000s, FlpC gained more attention as a promising compound by which to develop new antimicrobials to control human, animal, and plant pathogens. This study provides an overview of the results pertaining to the in vitro and in vivo antimicrobial activity of pure FlpC, as well as semi-purified fractions containing FlpC, against phytopathogenic microorganisms. Furthermore, the bioprospection history of the extensively researched FlpC-producing Pseudomonas aeruginosa LV strain and the related molecular evidence regarding the compound’s biosynthesis are discussed. Overall, FlpC is proposed to be an important alternative to antimicrobial resistance in human and animal health, and in tackling the negative environmental impacts caused by the exacerbated use of pesticides against phytopathogens.

In Vitro Antimelanoma Properties of Verbena officinalis Fractions.

Verbena officinalis is commonly used in traditional medicine to treat many ailments. Extracts of this plant are therapeutic agents for the potential treatment of different diseases, including colorectal and liver cancers, but have not been explored for their anti-melanoma potential so far. The goal of the current work was to prepare a methanolic extract and fractionate it using hexane, chloroform, ethyl acetate, butanol, and acetone to get semi-purified products. These semi-purified fractions were studied for their potency against melanoma cell lines. The three potent fractions (HA, VO79, and EA3) demonstrated 50% inhibition concentration (IC50) values as low as 2.85 µg/mL against the LOX IMVI cell line. All three fractions showed similar potency in inhibiting the growth of the B16 cells, a murine melanoma cell line. Based on high-resolution mass spectrometry (HRMS) data, for the first time, we report on lupulone A from this plant. LC-MS data also indicated the presence of hedergonic acid, serjanic acid, and other compounds in V. officinalis extracts.

Biological Assay-Guided Fractionation and Mass Spectrometry-Based Metabolite Profiling of Annona muricata L. Cytotoxic Compounds against Lung Cancer A549 Cell Line.

Annona muricata L. (Guyabano) leaves are reported to exhibit anticancer activity against cancer cells. In this study, the ethyl acetate extract from guyabano leaves was purified through column chromatography, and the cytotoxic effects of the semi-purified fractions were evaluated against A549 lung cancer cells using in vitro MTS cytotoxicity and scratch/wound healing assays. Fractions F15-16C and F15-16D exhibited the highest anticancer activity in the MTS assay, with % cytotoxicity values of 99.6% and 99.4%, respectively. The bioactivity of the fractions was also consistent with the results of the scratch/wound healing assay. Moreover, untargeted metabolomics was employed on the semi-purified fractions to determine the putative compounds responsible for the bioactivity. The active fractions were processed using LC-MS/MS analysis with the integration of the following metabolomic tools: MS-DIAL (for data processing), MetaboAnalyst (for data analysis), GNPS (for metabolite annotation), and Cytoscape (for network visualization). Results revealed that the putative compounds with a significant difference between active and inactive fractions in PCA and OPLS-DA models were pheophorbide A and diphenylcyclopropenone.

Purification, HR-LC-ESI-MS-MS Identification, and Peptide Prediction of Bacteriocin-Like Inhibitory Substances Produced by Streptomyces sp. Isolated from Chanos chanos.

Consumption of fresh and minimally processed food is closely related to foodborne diseases. To minimize the adverse effects, bacteriocin-like inhibitory substance (BLIS) as a natural preservative can be used. One of the bacteriocins with promising activity was produced by Streptomyces sp. Using gel filtration chromatography, the bacteriocin purification process succeeded in obtaining semi-purified fractions with broad-spectrum inhibitory activity to foodborne pathogen bacteria. These fractions are also stable up to 100 °C and pH 2.0–7.0. High-Resolution Liquid Chromatography Electrospray Ionization-Tandem Mass Spectrometry analysis followed by orthogonal projection to latent structure showed that each fraction had eight peaks with the highest positive correlation to BLIS-specific activity. Peptide identification based on MS spectrum found 597 predictive peptides, of which 42 predictive peptides with antimicrobial peptide characteristics and the highest iAMPpred antimicrobial peptide probability (>0.5) were selected. The selected predictive peptides have molecular mass of 247.13-615.37 Da and consist of at least 20% hydrophobic amino acids with a hydrophobicity value of 14.72 Kcal mol−1. The results of this study indicate the effectiveness of BLIS purification by gel filtration chromatography and the promising potential of semi-purified BLIS as a natural preservative. Besides, the active peptides in semi-purified BLIS can also be identified quickly so that the isolation process to obtain purified-BLIS can be carried out more efficiently.

α-Glucosidase, α-Amylase and Antioxidant Evaluations of Isolated Bioactives from Wild Strawberry.

Diabetes mellitus is a metabolic disorder and is a global challenge to the current medicinal chemists and pharmacologists. This research has been designed to isolate and evaluate antidiabetic bioactives from Fragaria indica. The crude extracts, semi-purified and pure bioactives have been used in all in vitro assays. The in vitro α-glucosidase, α-amylase and DPPH free radical activities have been performed on all plant samples. The initial activities showed that ethyl acetate (Fi.EtAc) was the potent fraction in all the assays. This fraction was initially semi-purified to obtain Fi.EtAc 1–3. Among the semi-purified fractions, Fi.EtAc 2 was dominant, exhibiting potent IC50 values in all the in vitro assays. Based on the potency and availability of materials, Fi.EtAc 2 was subjected to further purification to obtain compounds 1 (2,4-dichloro-6-hydroxy-3,5-dimethoxytoluene) and 2 (2-methyl-6-(4-methylphenyl)-2-hepten-4-one). The two isolated compounds were characterized by mass and NMR analyses. The compounds 1 and 2 showed excellent inhibitions against α-glucosidase (21.45 for 1 and 15.03 for 2 μg/mL), α-amylase (17.65 and 16.56 μg/mL) and DPPH free radicals (7.62 and 14.30 μg/mL). Our study provides baseline research for the antidiabetic bioactives exploration from Fragaria indica. The bioactive compounds can be evaluated in animals-based antidiabetic activity in future.

Identification of Antimycobacterial Natural Products from a Library of Marine Invertebrate Extracts

Tuberculosis (TB) remains a public health crisis, requiring the urgent identification of new anti-mycobacterial drugs. We screened several organic and aqueous marine invertebrate extracts for their in vitro inhibitory activity against the causative organism, Mycobacterium tuberculosis. Here, we report the results obtained for 54 marine invertebrate extracts. The chemical components of two of the extracts were dereplicated, using 1H NMR and HR-LCMS with GNPS molecular networking, and these extracts were further subjected to an activity-guided isolation process to purify the bioactive components. Hyrtios reticulatus yielded heteronemin 1 and Jaspis splendens was found to produce the bengamide class of compounds, of which bengamides P 2 and Q 3 were isolated, while a new derivative, bengamide S 5, was putatively identified and its structure predicted, based on the similarity of its MS/MS fragmentation pattern to those of other bengamides. The isolated bioactive metabolites and semi-pure fractions exhibited M. tuberculosis growth inhibitory activity, in the range <0.24 to 62.50 µg/mL. This study establishes the bengamides as potent antitubercular compounds, with the first report of whole-cell antitubercular activity of bengamides P 2 and Q 3.

Encapsulation of phlorotannins from edible brown seaweed in chitosan: Effect of fortification on bioactivity and stability in functional foods

Phlorotannins are a family of proven therapeutic agents. However, low stability disturbs their full bioactivity expression in the human body. Hence, this study focused on preserving their vitality through encapsulation. Phlorotannins isolated from Sargassum ilicifolium were encapsulated in the chitosan-tripolyphosphate carrier. Their storage stability, processing stability, and bioactivity retention upon in vitro digestion were determined. Results revealed the highest total phlorotannin content (TPC) of 854.38 ± 48 mg Phloroglucinol Equivalence/g in the semi-purified ethyl acetate fraction while the NMR spectrum and the LCMS profile revealed the isolation of phlorotannins in it. Storage at –18℃ and 4℃ temperatures preserved thrice both the encapsulated and non-encapsulated phlorotannins than ambient conditions. Encapsulated compound reported 56.4% of TPC retention at 175 ℃ processing temperature. Fermented fraction of encapsulated form showed significantly higher (p < 0.05) antioxidant activities and TPC (0.23 ± 0.03 mg/mL) suggesting the potential for targeted delivery of phlorotannins to their absorption sites through encapsulation.

Cytotoxicity of extracts and compounds isolated from Croton echioides in animal tumor cell (HTC).

The search for compounds with anticancer effects is of paramount importance today due to the high incidence of the disease. The Euphorbiaceae family is known for having compounds with therapeutic properties, one of its genera being Croton. It has several species, which contain compounds already known for their biological activities, presenting anti-inflammatory, antimicrobial and anticancer properties. Thus, the cytotoxicity/antiproliferative activity of semi-purified fractions and compounds isolated from Croton echioides in liver tumor cells of Rattus norvegicus (HTC) was evaluated by the MTT test. The semi-purified fractions showed cytotoxicity at concentrations above 200 µg/mL, at 24, 48 and 72 hours, reaching cell viability of 24.78% [400 µg/mL] at 24 hours, 12.79% [500 µg/mL] at 48 hours and 10.57% [300 µg/mL] at 72 hours. For the isolated compounds, lupeol had a cytotoxic effect in all concentrations (1, 5, 10, 15, 20, 40, 60, 80 and 100 µg/mL) and tested times (24, 48 and 72 hours), reaching minimum viability of 4.37% [100 µg/mL], within 72 hours. The clerodan diterpenes CEH-1 and CEH-4 also showed antiproliferative activity, with minimum viability of 36.19% [100 µg/mL] over 72 hours and 21.33% [100 µg/mL] over 48 hours, respectively. However, the clerodan diterpenes CEH-2 and CEH-3 did not shows a cytotoxic effect for HTC cells. Thus, there is a cytotoxic/antiproliferative potential of C. echioides against tumor cells, with targeted to mitochondrial enzymes, associated with cell proliferation, indicating that this species deserves prominence in the search for new molecules for the treatment of cancer.