Next Article in Journal
Fabrication of Cr-ZnFe2O4/S-g-C3N4 Heterojunction Enriched Charge Separation for Sunlight Responsive Photocatalytic Performance and Antibacterial Study
Next Article in Special Issue
Pickering Emulsions as Vehicles for Bioactive Compounds from Essential Oils
Previous Article in Journal
Erianin-Loaded Photo-Responsive Dendrimer Mesoporous Silica Nanoparticles: Exploration of a Psoriasis Treatment Method
Previous Article in Special Issue
Synthesis and Investigation of Flavanone Derivatives as Potential New Anti-Inflammatory Agents
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Molecules
Volume 27
Issue 19
10.3390/molecules27196329
molecules-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Communication

In Vitro Antimelanoma Properties of Verbena officinalis Fractions

by
Rabia Nisar
1,
Sanjay Adhikary
2,
Saeed Ahmad
1 and
Mohammad Abrar Alam
2,*
1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan
2
Department of Chemistry and Physics, College of Sciences and Mathematics, Arkansas State University, Jonesboro, AR 72404, USA
*
Author to whom correspondence should be addressed.
Molecules 2022, 27(19), 6329; https://doi.org/10.3390/molecules27196329
Submission received: 20 August 2022 / Revised: 18 September 2022 / Accepted: 21 September 2022 / Published: 26 September 2022
(This article belongs to the Special Issue Sustainable Approaches to Natural Product-Inspired Drug Discovery and Development)
Browse Figures
Review Reports Versions Notes

Abstract

:
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.

1. Introduction

Humans have been utilizing plants therapeutically for a long time. The World Health Organization (WHO) estimates that almost 80% of the world’s population relies on medicinal plants to cover their basic healthcare needs [1]. Traditional medications made from plants are considered more effective and safer clinically when compared to synthetic entities [2]. Approximately 25% of the medications prescribed worldwide are originated from plants [3]. Medicinal plants containing a rich source of bioactive compounds are the most common source of novel drug discovery and are particularly useful as antimicrobial and anticancer therapeutics [4]. A recent analysis found that around 50% of approved anticancer drugs between 1940 to 2014 were obtained from natural sources or directly derived from them [5].
Verbena officinalis, a plant from the Verbenaceae family, is commonly called pigeons’ grass, herb of grace, or vervain. This plant is popularly called bitter herb or Kori-booti in Pakistan. It is mostly found in Asia, North Africa, and Europe. This plant is fairly distributed near water in cultivated fields and wastelands in the western and northern regions of Pakistan. It is a perennial erect herb, which grows to a height of about 25–100 cm, having toothed and lobed leaves. The delicate spikes hold elegant, silky, pale purple, or pink flowers [6]. In the traditional herbal system of medicines, V. officinalis has been employed for the treatment of many ailments such as gastric diseases, abrasion, skin burns [7], wounds, thyroid problems, rheumatic pain [8], asthma and cough [9], amenorrhea, enteritis, acute dysentery [10], expectorant and diuretic [11]. V. officinalis studied for its new important bioactivities including analgesic and anti-inflammatory [12,13], antioxidant [14], antifungal [15], anticonvulsant [6], antibacterial [16,17], anticancer [18,19], neuroprotective [10], antidepressant [20], antiproliferative [21], urolithiasis [22], and antitumor [23] effects. V. officinalis has a pool of bioactive metabolites, including flavonoids [24], sterols and triterpenoids [25], phenylethanoid glycosides [21], iridoids [26] and ursolic acid [27], which further explains the folk use of this plant [6]. In addition, the species nowadays is recognized as a valuable cosmetic plant, mainly due to the presence of essential oils. The vervain herb is characterized by high variability in chemical composition depending on its origin [28].
Melanoma is the most fatal and aggressive skin cancer that accounts for 3% of all malignant cancer [29]. Melanoma has a high potential for metastasis and invasion, which accounts for about 75% of death related to skin cancer worldwide [30]. It is the 5th and 7th most prevalent cancer in American men and women, respectively [31]. In the United States, the expected number of new cases and deaths in 2022 will be 99,780 and 7650, respectively [32]. Antimelanoma properties of plant extracts such as Aloysia citrodora essential oil inhibit melanoma cell growth and migration by targeting HB-EGF-EGFR signaling [33]. The aim of this study was to analyze the important secondary metabolites of V. officinalis for the evaluation of their antimelanoma potential. Until now, this is the first anti-melanoma study of V. officinalis extracts.

2. Results and Discussion

2.1. In Vitro Antimelanoma Studies of Different Fractions

Initially, we tested the different extracts of V. officinalis at 50 µg/mL against the LOX IMVI cells, a melanoma cell line. Acetone and n-butanol fractions did not show any cytotoxicity for this melanoma cell line at this concentration (Figure 1). Therefore, we did not pursue these two fractions of the methanolic extract of V. officinalis further. The extracts of hexane, chloroform, and ethyl acetate showed >70% growth inhibition at 50 µg/mL concentration. We further fractionated the extract of chloroform by column chromatography.
All these fractions were tested against LOX IMVI cell line at 50 µg/mL concentration. Extracts showing >70% growth inhibition were tested against the cell lines at five serial dilutions. We found three potent fractions (HA, VO79, and EA3) with 50% inhibition concentration (IC50) as low as 2.8 µg/mL against the LOX IMVI cell line (Table 1). The fraction VO79 inhibited the growth of melanoma cell lines (SK MEL 28, LOX IMVI, and SK MEL 5) with the IC50 values in the range of 6.2 to 11.6 µg/mL. This fraction inhibited the growth of a murine melanoma cell line (B16 cell line) with an IC50 value of 7.0 µg/mL. The HA fraction was found to be the most potent isolate of the V. officinalis to inhibit the growth of LOX IMVI cell lines. The EA3 fraction was very efficient in inhibiting the growth of SK-MEL-28 and LOX IMVI cell lines with the IC50 values of 4.8 and 3.3 µg/mL, respectively. All three fractions inhibited the growth of the B16 cell line effectively, with IC50 values as low as 6.2 µg/mL. These results are very significant as the positive controls is 3–4 times less potent than these fractions against the melanoma cell lines except for LOX IMVI cell line against cisplatin.

2.2. Phytochemical Analysis of V. officinalis by HR-ESI-MS

The three potent fractions were subjected to HR-EST-MS analysis. The data were compared with online databases, such as NIST Chemistry WebBook and PubChem. The structure of 13 metabolites of V. officinalis belonging to different phytochemical groups was putatively assigned using mass spectrometry (Table 2). We have found the bis(2-ethylhexyl)phthalate and lupulone A using high-resolution mass spectrometry (HRMS) data (Supplementary Materials). This is the first report indicating the presence of lupolone A in this plant. This result is very significant, as lupulones are known to show anticancer properties [34]. Triterpenoids were the representative chemical class consisting of seven compounds—hederagonic or glycyrrhetinic acid (V4), (2α,3β)-2,3-dihydroxyurs-12-en-28-oic acid or hederagenin (V5), gypensapogenin A (V6), momordicinin (V8), camarolide (V9), ursonic or moronic acid (V10), and serjanic acid (V12). Three compounds (V3, V11, and V13) did not match the structures of the online database.

3. Materials and Methods

3.1. Chemicals, Instruments, and Software

Chemicals and silica: Methanol, n-hexane, chloroform, ethyl acetate, n-butanol, normal phase silica gel mesh size (70–230); Instrument: preparative TLC columns, Rotary evaporator, Shimadzu IT-TOF mass spectrometer, CytationTM5 (BioTek, Winooski, VT, USA), and GraphPad Prism 9 (software).

3.2. Collection of Plant Materials

The whole plant of V. officinalis was collected during the flowering season, November 2017 to March 2018, from tehsil Gojra, district Toba Tek Singh Pakistan (Figure 2). The taxonomic status of the plant was verified by Dr. Zaheer-ud-Din Khan, Government College University, Lahore, Pakistan. A voucher number 3514 of the plant specimen was deposited in the department of Botany, Government College University, Lahore, Pakistan.

3.3. Extraction and Fractionation

A whole plant was shade dried and then ground into a coarse powder. Pulverized powder (10 kg) was macerated in 80% aqueous methanol for two weeks under normal conditions with occasional shaking. The methanolic extract was filtered and concentrated at 40 °C in a vacuum using a rotary evaporator under reduced pressure to yield a dry crude extract (460 g). The dry methanolic extract of V. officinalis was suspended in 1 L of distilled water and extracted with hexane, chloroform, ethyl acetate, n-butanol and acetone successively to obtain different solvent fractions. Each fraction was then concentrated by using a rotary evaporator and weighed. All extracts were then stored in sealed containers in a refrigerator for further purification and biological evaluation.

3.3.1. Hexane Fraction

After fractionating the methanolic crude extract using hexane solvent, 2 g of hexane fraction (HA) was produced as a yellow color oil.

3.3.2. Isolation of EA3 from the Ethyl Acetate Fraction

The ethyl acetate fraction (100 g) was obtained by performing extraction of the crude methanolic extract with ethyl acetate solvent. A greenish powder was obtained from the ethyl acetate extract, which was recrystallized from methanol to obtain a white powder (EA3).

3.3.3. Isolation of VO79 from the Chloroform Fraction with Column Chromatography

The chloroform fraction (80 g) was subjected to column chromatography using a wet technique, and elution was started with hexane, chloroform, and chloroform-methanol solvent system. Elution was started at pure hexane, and the polarity was increased by 10% by adding chloroform until it reached 100% chloroform. The polarity was increased gradually by adding methanol with a 10% increment in the solvent system until 100% methanol. This whole process has resulted in the elution of 21 sub-fractions. The fractions eluted at the solvent system chloroform: methanol (9:1 to 7:3) was obtained in a significant quantity (11 g). It was again subjected to column chromatography. VO79 fraction was eluted with hexane: chloroform (1:9 to 0:1) solvent system with a 2% polarity increment.

3.3.4. N-Butanol Fraction

V. officinalis dried methanolic extract fractionated using n-butanol as the solvent, yielding a 95 g n-butanol fraction.

3.3.5. Acetone Fraction

Dried methanolic extract was fractionated with acetone solvent, affording an acetone fraction (29 g).

3.4. Cytotoxicity Studies

The Resazurin assay was performed to assess the cytotoxicity of the compounds as we described previously [48,49]. A 96-well plate was seeded with 6000 cells/well and incubated for 24 h in the presence of growth media. After 24 h of incubation, cells were treated with six two-fold serial dilutions of fractions from 25 (µM) to 0.78 µM. Each dilution was treated in triplicate, and another 24 h incubation was performed. After 24 h of treatment, resazurin dye was added at a final concentration of 25 µg/mL, and again the cells were incubated for 4 h. Finally, the fluorescence intensity was measured at 560 nm excitation and 590 nm emission using CytationTM 5 (BioTek, Winooski, VT, USA). The non-linear regression of viable cells and treatment concentration was performed using a GraphPad Prism 9 (San Diego, CA, USA) to calculate the 50% inhibition concentration (IC50) of fractions.

4. Conclusions

For the first time, we have revealed the anti-melanoma properties of V. officinalis extracts. These mixtures of compounds are potent growth inhibitors in different melanoma cell lines with IC50 values at low micromolar concentrations. These potent fractions are very significant as these fractions of multiple compounds can be further separated into individual compounds, which can be significantly more potent than the mixtures. We have putatively characterized these fractions by using HRMS. The potent cytotoxic properties of these fractions warrant further separation and anti-melanoma studies of V. officinalis extracts.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/molecules27196329/s1, Figure S1: ESI high-resolution mass spectrum of HA extract of V. officinalis; Figure S2. LC chromatogram of VO79 isolated from the chloroform extract of V. officinalis; Figure S3. ESI mass spectrum of VO79 at retention time 3.37 min; Figure S4. ESI mass spectrum of VO79 at retention time 8.21 min; Figure S5. ESI mass spectrum of VO79 at retention time 9.53 min; Figure S6. LC chromatogram of EA3 isolated from the ethyl acetate extract of V. officinalis; Figure S7. ESI mass spectrum of EA3 at retention time 5.41 min; Figure S8. ESI mass spectrum of EA3 at retention time 5.58 min; Figure S9. ESI mass spectrum of EA3 at retention time 6.23 min; Figure S10. ESI mass spectrum of EA3 at retention time 6.52 min; Figure S11. ESI mass spectrum of EA3 at retention time 6.86 min; Figure S12. ESI mass spectrum of EA3 at retention time 7.13 min; Figure S13. ESI mass spectrum of EA3 at retention time 7.55 min; Figure S14. ESI mass spectrum of EA3 at retention time 7.55 min; Figure S15. ESI mass spectrum of EA3 at retention time 8.23 min; Figure S16. ESI mass spectrum of EA3 at retention time 8.59 min; Figure S17. ESI mass spectrum of EA3 at retention time 10.23 min

Author Contributions

S.A. (Saeed Ahmad) and R.N. selected the plant and extracted the compounds/fractions, and M.A.A. got the mass spectrometry data and conceptualized the anti-melanoma studies. R.N. wrote the manuscript and M.A.A. edited it for the final draft. S.A. (Sanjay Adhikary) carried out the cytotoxicity studies. All authors have read and agreed to the published version of the manuscript.

Funding

This publication was made possible by the Research Technology Core of the Arkansas INBRE program, supported by a grant from the National Institute of General Medical Sciences, (NIGMS), P20 GM103429 from the National Institutes of Health.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Acknowledgments

The authors would like to thank the Arkansas INBRE program, supported by a grant from the National Institute of General Medical Sciences (NIGMS), P20 GM103429 from the National Institutes of Health for the mass spectrometry data, and the Arkansas Biosciences Institute for the infrastructure for cytotoxicity studies. Rabia Nisar would like to thank International Research Support Initiative Program by Higher Education Commission Pakistan.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Akther, Y.; Nabi, J.; Tabassum, N. Comprehensive Overview of Some Edible Medicinal Plants from Kashmir Valley: Cultural, Economic, and Pharmacological Importance. In Edible Plants in Health and Diseases; Springer: Singapore, 2022; pp. 137–159. [Google Scholar]
  2. Aljohny, B.O.; Rauf, A.; Anwar, Y.; Naz, S.; Wadood, A. Antibacterial, Antifungal, Antioxidant, and Docking Studies of Potential Dinaphthodiospyrols from Diospyros lotus Linn Roots. ACS Omega 2021, 6, 5878–5885. [Google Scholar] [CrossRef] [PubMed]
  3. Sahoo, N.; Manchikanti, P.; Dey, S. Herbal drugs: Standards and regulation. Fitoterapia 2010, 81, 462–471. [Google Scholar] [CrossRef] [PubMed]
  4. Cho, H.; Shen, Q.; Zhang, L.H.; Okumura, M.; Kawakami, A.; Ambrose, J.; Sigoillot, F.; Miller, H.R.; Gleim, S.; Cobos-Correa, A. CYP27A1-dependent anti-melanoma activity of limonoid natural products targets mitochondrial metabolism. Cell Chem. Biol. 2021, 28, 1407–1419.e6. [Google Scholar] [CrossRef]
  5. Newman, D.J.; Cragg, G.M. Natural products as sources of new drugs from 1981 to 2014. J. Nat. Prod. 2016, 79, 629–661. [Google Scholar] [CrossRef] [PubMed]
  6. Khan, A.W.; Khan, A.-U.; Ahmed, T. Anticonvulsant, anxiolytic, and sedative activities of Verbena officinalis. Front. Pharmacol. 2016, 7, 499. [Google Scholar] [CrossRef]
  7. Speroni, E.; Cervellati, R.; Costa, S.; Guerra, M.; Utan, A.; Govoni, P.; Berger, A.; Müller, A.; Stuppner, H. Effects of differential extraction of Verbena officinalis on rat models of inflammation, cicatrization and gastric damage. Planta Med. 2007, 73, 227–235. [Google Scholar] [CrossRef]
  8. Guarrera, P.M.; Forti, G.; Marignoli, S. Ethnobotanical and ethnomedicinal uses of plants in the district of Acquapendente (Latium, Central Italy). J. Ethnopharmacol. 2005, 96, 429–444. [Google Scholar] [CrossRef]
  9. Vitalini, S.; Tomè, F.; Fico, G. Traditional uses of medicinal plants in Valvestino (Italy). J. Ethnopharmacol. 2009, 121, 106–116. [Google Scholar] [CrossRef]
  10. Lai, S.-W.; Yu, M.-S.; Yuen, W.-H.; Chang, R.C.-C. Novel neuroprotective effects of the aqueous extracts from Verbena officinalis Linn. Neuropharmacology 2006, 50, 641–650. [Google Scholar] [CrossRef]
  11. Schönbichler, S.; Bittner, L.; Pallua, J.; Popp, M.; Abel, G.; Bonn, G.; Huck, C. Simultaneous quantification of verbenalin and verbascoside in Verbena officinalis by ATR-IR and NIR spectroscopy. J. Pharm. Biomed. Anal. 2013, 84, 97–102. [Google Scholar] [CrossRef]
  12. Calvo, M. Anti-inflammatory and analgesic activity of the topical preparation of Verbena officinalis L. J. Ethnopharmacol. 2006, 107, 380–382. [Google Scholar] [CrossRef] [PubMed]
  13. Calvo, M.; Vilalta, N.; San Julian, A.; Fernandez, M. Anti-inflammatory activity of leaf extract of Verbena officinalis L. Phytomedicine 1998, 5, 465–467. [Google Scholar] [CrossRef]
  14. Bilia, A.; Giomi, M.; Innocenti, M.; Gallori, S.; Vincieri, F. HPLC-DAD-ESI-MS analysis of the constituents of aqueous preparations of verbena and lemon verbena and evaluation of the antioxidant activity. J. Pharm. Biomed. Anal. 2008, 46, 463–470. [Google Scholar] [CrossRef] [PubMed]
  15. Casanova, E.; GarcAa-Mina, J.; Calvo, M. Antioxidant and antifungal activity of Verbena officinalis L. leaves. Plant Foods Hum. Nutr. 2008, 63, 93–97. [Google Scholar] [CrossRef]
  16. Hernández, N.E.; Tereschuk, M.; Abdala, L. Antimicrobial activity of flavonoids in medicinal plants from Tafı del Valle (Tucuman, Argentina). J. Ethnopharmacol. 2000, 73, 317–322. [Google Scholar] [CrossRef]
  17. Mengiste, B.; Lulie, S.; Getachew, B.; Gebrelibanos, M.; Mekuria, A.; Masresha, B. In vitro antibacterial activity of extracts from aerial parts of Verbena officinalis. Adv. Biol. Res. 2015, 9, 53–57. [Google Scholar]
  18. Martino, L.D.; D’Arena, G.; Minervini, M.M.; Deaglio, S.; Sinisi, N.P.; Cascavilla, N.; Feo, V.D. Active caspase-3 detection to evaluate apoptosis induced by Verbena officinalis essential oil and citral in chronic lymphocytic leukaemia cells. Rev. Bras. Farmacogn. 2011, 21, 869–873. [Google Scholar] [CrossRef]
  19. Martino, L.d.; D’Arena, G.; Minervini, M.; Deaglio, S.; Fusco, B.; Cascavilla, N.; Feo, V.D. Verbena officinalis essential oil and its component citral as apoptotic-inducing agent in chronic lymphocytic leukemia. Int. J. Immunopathol. Pharmacol. 2009, 22, 1097–1104. [Google Scholar] [CrossRef]
  20. Bekara, A.; Amazouz, A.; Douma, T.B. Evaluating the antidepressant Effect of Verbena officinalis L.(Vervain) aqueous extract in adult rats. Basic Clin. Neurosci. 2020, 11, 91. [Google Scholar] [CrossRef]
  21. Encalada, M.A.; Rehecho, S.; Ansorena, D.; Astiasaran, I.; Cavero, R.Y.; Calvo, M.I. Antiproliferative effect of phenylethanoid glycosides from Verbena officinalis L. on colon cancer cell lines. LWT-Food Sci. Technol. 2015, 63, 1016–1022. [Google Scholar] [CrossRef]
  22. Grases, F.; Melero, G.; Costa-Bauza, A.; Prieto, R.; March, J. Urolithiasis and phytotherapy. Int. Urol. Nephrol. 1994, 26, 507–511. [Google Scholar] [CrossRef] [PubMed]
  23. Kou, W.-Z.; Yang, J.; Yang, Q.-H.; Wang, Y.; Wang, Z.-F.; Xu, S.-L.; Liu, J. Study on in-vivo anti-tumor activity of Verbena officinalis extract. Afr. J. Tradit. Complement. Altern. Med. 2013, 10, 512–517. [Google Scholar] [CrossRef] [PubMed]
  24. Calvo, M.; San Julian, A.; Fernandez, M. Identification of the major compounds in extracts of Verbena officinalis L. (Verbenaceae) by HPLC with post-column derivatization. Chromatographia 1997, 46, 241–244. [Google Scholar] [CrossRef]
  25. Deepak, M.; Handa, S.S. Antiinflammatory activity and chemical composition of extracts of Verbena officinalis. Phytother. Res. 2000, 14, 463–465. [Google Scholar] [CrossRef]
  26. Rimpler, H.; Schafer, B. Hastatoside, a new iridoid from Verbena hastata L. and Verbena officinalis L. Z. Far Nat. C 1979, 34, 311–318. [Google Scholar]
  27. Kaur, J.; Kumar, D.; Madaan, R.; Kumar, S. Estimation of isolated triterpenoid-ursolic acid in Verbena officinalis L. aerial parts using TLC densitometry. J. Pharm. Technol. Res. Manag. 2014, 2, 121–135. [Google Scholar] [CrossRef]
  28. Kubica, P.; Szopa, A.; Kokotkiewicz, A.; Miceli, N.; Taviano, M.F.; Maugeri, A.; Cirmi, S.; Synowiec, A.; Gniewosz, M.; Elansary, H.O. Production of Verbascoside, Isoverbascoside and phenolic acids in callus, suspension, and bioreactor cultures of Verbena officinalis and biological properties of biomass extracts. Molecules 2020, 25, 5609. [Google Scholar] [CrossRef]
  29. AlQathama, A.; Prieto, J. Natural products with therapeutic potential in melanoma metastasis. Nat. Prod. Rep. 2015, 32, 1170–1182. [Google Scholar] [CrossRef]
  30. Jin, S.; Kim, K.C.; Kim, J.-S.; Jang, K.-I.; Hyun, T.K. Anti-melanoma activities and phytochemical compositions of sorbus commixta fruit extracts. Plants 2020, 9, 1076. [Google Scholar] [CrossRef]
  31. Chambers, S.A.; Newman, M.; Frangie, M.M.; Savenka, A.V.; Basnakian, A.G.; Alam, M.A. Antimelanoma activities of chimeric thiazole–androstenone derivatives. R. Soc. Open Sci. 2021, 8, 210395. [Google Scholar] [CrossRef]
  32. National Cancer Institute. Cancer Stat Facts: Melanoma of the Skin. Available online: https://seer.cancer.gov/statfacts/html/melan.html (accessed on 15 August 2022).
  33. Salama, Y.; Jaradat, N.; Hattori, K.; Heissig, B. Aloysia Citrodora Essential Oil Inhibits Melanoma Cell Growth and Migration by Targeting HB-EGF-EGFR Signaling. Int. J. Mol. Sci. 2021, 22, 8151. [Google Scholar] [CrossRef] [PubMed]
  34. Tyrrell, E.; Archer, R.; Tucknott, M.; Colston, K.; Pirianov, G.; Ramanthan, D.; Dhillon, R.; Sinclair, A.; Skinner, G.A. The synthesis and anticancer effects of a range of natural and unnatural hop β-acids on breast cancer cells. Phytochem. Lett. 2012, 5, 144–149. [Google Scholar] [CrossRef]
  35. Habib, M.R.; Karim, M.R. Antitumour evaluation of di-(2-ethylhexyl) phthalate (DEHP) isolated from Calotropis gigantea L. flower. Acta Pharm. 2012, 62, 607–615. [Google Scholar] [CrossRef] [PubMed]
  36. Yu, J.-H.; Yu, Z.-P.; Wang, Y.-Y.; Bao, J.; Zhu, K.-K.; Yuan, T.; Zhang, H. Triterpenoids and triterpenoid saponins from Dipsacus asper and their cytotoxic and antibacterial activities. Phytochemistry 2019, 162, 241–249. [Google Scholar] [CrossRef]
  37. Kao, T.-C.; Wu, C.-H.; Yen, G.-C. Bioactivity and Potential Health Benefits of Licorice. J. Agric. Food Chem. 2014, 62, 542–553. [Google Scholar] [CrossRef]
  38. Woo, H.J.; Lee, J.Y.; Woo, M.H.; Yang, C.H.; Kim, Y.H. Apoptogenic activity of 2α,3α-dihydroxyurs-12-ene-28-oic acid from Prunella vulgaris var. lilacina is mediated via mitochondria-dependent activation of caspase cascade regulated by Bcl-2 in human acute leukemia Jurkat T cells. J. Ethnopharmacol. 2011, 135, 626–635. [Google Scholar] [CrossRef]
  39. Shu, J.-C.; Liu, J.-Q.; Chou, G.-X. A new triterpenoid from Verbena officinalis L. Nat. Prod. Res. 2013, 27, 1293–1297. [Google Scholar] [CrossRef]
  40. Attaur, R.; Ansari, A.A.; Kenne, L. Hederagenin, Ursolic Acid, and Pinatol from Fagonia indica. J. Nat. Prod. 1984, 47, 186–187. [Google Scholar]
  41. Li, N.; Wu, C.-F.; Xu, X.-Y.; Liu, Z.-Y.; Li, X.; Zhao, Y.-Q. Triterpenes possessing an unprecedented skeleton isolated from hydrolyzate of total saponins from Gynostemma pentaphyllum. Eur. J. Med. Chem. 2012, 50, 173–178. [Google Scholar] [CrossRef]
  42. Aftab, Z.; Afzal, M.; Bushra; Khan, H.; Badshah, S.; Khan, D.; Ullah, H.; Khan, S. Fistuloates A–C: New antioxidative aromatic compounds isolated from Cassia fistula. J. Chem. Res. 2019, 43, 516–521. [Google Scholar] [CrossRef]
  43. Serala, K.; Steenkamp, P.; Mampuru, L.; Prince, S.; Poopedi, K.; Mbazima, V. In vitro antimetastatic activity of Momordica balsamina crude acetone extract in HT-29 human colon cancer cells. Environ. Toxicol. 2021, 36, 2196–2205. [Google Scholar] [CrossRef] [PubMed]
  44. Lenora, L.; Kumar, J.S.; Murugesan, S.; Senthilkumar, N. GGC-MS-MS analysis of alien invasive aquatic weed, Eichhornia crassipes (Mart.) Solms. Chem. Sin. 2016, 7, 48–52. [Google Scholar]
  45. Van der Doelen, G.A.; van den Berg, K.J.; Boon, J.J.; Shibayama, N.; De La Rie, E.R.; Genuit, W.J.L. Analysis of fresh triterpenoid resins and aged triterpenoid varnishes by high-performance liquid chromatography–atmospheric pressure chemical ionisation (tandem) mass spectrometry. J. Chromatogr. A 1998, 809, 21–37. [Google Scholar] [CrossRef]
  46. Rios, M.Y.; Salinas, D.; Villarreal, M.L. Cytotoxic Activity of Moronic Acid and Identification of the New Triterpene 3,4-seco-Olean-18-ene-3,28-dioic Acid from Phoradendron reichenbachianum. Planta Med. 2001, 67, 443–446. [Google Scholar] [CrossRef] [PubMed]
  47. Peláez, G.L.M.; Sierra, J.A.; Alzate, F.; Holzgrabe, U.; Ramirez-Pineda, J.R. Pentacyclic triterpenes from Cecropia telenitida with immunomodulatory activity on dendritic cells. Rev. Bras. Farmacogn. 2013, 23, 754–761. [Google Scholar] [CrossRef]
  48. Alkhaibari, I.S.; Raj, K.C.H.; Alnufaie, R.; Gilmore, D.; Alam, M.A. Synthesis of Chimeric Thiazolo-Nootkatone Derivatives as Potent Antimicrobial Agents. ChemMedChem 2021, 16, 2628–2637. [Google Scholar] [CrossRef]
  49. Hansa, R.K.C.; Khan, M.M.K.; Frangie, M.M.; Gilmore, D.F.; Shelton, R.S.; Savenka, A.V.; Basnakian, A.G.; Shuttleworth, S.L.; Smeltzer, M.S.; Alam, M.A. 4-4-(Anilinomethyl)-3-[4-(trifluoromethyl)phenyl]-1H-pyrazol-1-ylbenzoic acid derivatives as potent anti-gram-positive bacterial agents. Eur. J. Med. Chem. 2021, 219, 113402. [Google Scholar] [CrossRef]
Molecules 27 06329 g001 550
Figure 1. Flow chart for the melanoma active fractions of V. officinalis.
Figure 1. Flow chart for the melanoma active fractions of V. officinalis.
Molecules 27 06329 g001
Molecules 27 06329 g002 550
Figure 2. Verbena officinalis (06-21-2018, Chak NO. 363 JB, Tehsil Gojra, District Toba Tek Singh, Division Faisalabad 56000).
Figure 2. Verbena officinalis (06-21-2018, Chak NO. 363 JB, Tehsil Gojra, District Toba Tek Singh, Division Faisalabad 56000).
Molecules 27 06329 g002
Table
Table 1. IC50 values of the HA, VO79, EA3 fractions (µg/mL) against murine (B16) and human (SK MEL 28, LOX IMVI, and SK MEL 5) melanoma cell lines. NB: IC50 values are presented with mean ± Standard deviation. Taxol and cisplatin are positive controls.
Table 1. IC50 values of the HA, VO79, EA3 fractions (µg/mL) against murine (B16) and human (SK MEL 28, LOX IMVI, and SK MEL 5) melanoma cell lines. NB: IC50 values are presented with mean ± Standard deviation. Taxol and cisplatin are positive controls.
FractionIC50 (µg/mL)
B16SK MEL 28LOX IMVISK MEL 5
HA7.6± 1.110.6 ± 1.02.8 ± 0.28.0 ± 0.1
VO797.0 ± 0.87.2 ± 0.76.2 ± 0.19.6 ± 0.0
EA36.2 ± 0.84.8 ± 1.03.3 ± 0.06.2 ± 0.1
Taxol (µM)27.4 ± 4.527.3 ± 2.8132.1 ± 1.119.8 ± 3.1
Cisplatin24.3 ± 1.0 5.4 ± 0.327.1 ± 3.1
Table
Table 2. Tentative identification of the phytochemicals of V. officinalis extracts.
Table 2. Tentative identification of the phytochemicals of V. officinalis extracts.
NoProbable CompoundsMFm/z (found)ClassStructurem/z (cald)Ref.
HAV1Bis(2-ethylhexyl) phthalateC24H38O4391.2812Phthalate Molecules 27 06329 i001391.2842[35]
V2Lupulone AC26H36O4413.2665β-Bitter acids Molecules 27 06329 i002413.2686[34]
Vo79V3UnknownC26H30O3391.23----
V4Hederagonic acid
or
Glycyrrhetinic acid		C30H46O4471.34Triterpenoids 471.3468[36,37]
V5(2α,3β)-2,3-Dihydroxyurs-12-en-28-oic acid
or
Hederagenin		C30H48O4473.36Triterpenoids 473.3625[38]
[39,40]
EA3V6Gypensapogenin AC30H42O2435.32Triterpenes Molecules 27 06329 i003435.3257[41]
V7Fistuloate AC30H44O2437.34Aromatic compounds Molecules 27 06329 i004436.67[42]
V8MomordicininC30H46O2439.35Triterpenoids Molecules 27 06329 i005439.3570[43]
V9CamarolideC30H44O3453.33Triterpenoids Molecules 27 06329 i006453.3363[44]
V10Ursonic acid
or
Moronic acid		C30H46O3455.35Triterpenoids 455.3519[45,46]
V11UnknownC31H48O4485.36----
V12Serjanic acidC31H48O5501.36Pentacyclic triterpenes Molecules 27 06329 i007501.3574[47]
V13UnknownC34H46O5511.34----
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Nisar, R.; Adhikary, S.; Ahmad, S.; Alam, M.A. In Vitro Antimelanoma Properties of Verbena officinalis Fractions. Molecules 2022, 27, 6329. https://doi.org/10.3390/molecules27196329

AMA Style

Nisar R, Adhikary S, Ahmad S, Alam MA. In Vitro Antimelanoma Properties of Verbena officinalis Fractions. Molecules. 2022; 27(19):6329. https://doi.org/10.3390/molecules27196329

Chicago/Turabian Style

Nisar, Rabia, Sanjay Adhikary, Saeed Ahmad, and Mohammad Abrar Alam. 2022. "In Vitro Antimelanoma Properties of Verbena officinalis Fractions" Molecules 27, no. 19: 6329. https://doi.org/10.3390/molecules27196329

Article Metrics

Back to TopTop