In silico design of Antimalarial drug from Catharanthus roseus (G. Don)alkaloid molecules against AMA1 protein

Introduction          

Malaria keeps on causing unsuitably significant degrees of illness and demise, as archived in progressive versions of the World Malaria report.¹ As indicated by the most recent report, there were an expected 627000 lives lost in malaria 2020.² Malaria is preventable and treatable and the worldwide need is to lessen the weight of infection and demise while holding the drawn out vision of Malaria destruction.

The only malaria vaccine, RTS,S/AS01, initiates incomplete adequacy through enlistment of antibodies against the sequence (Asn-Ala-Asn-Pro) of the circumsporozoite protein (CSP). However, there are many antigenic determinants of Plasmodium against which either medications or antibodies are formed. ³ The different standards for antigen choice as well as medication definition are summed up in table.1.

Table 1: Plasmodium exposed surface antigens are:

The current situation is confronting many difficulties in vector control and parasite disposal. One of the most significant challenges is drug obstruction in Plasmodium. While obstruction is regularly evaluated by drug adequacy concentrates on that uncover applicant point transformations, whose pervasiveness are checked. The genuine method of activity of obstruction is related with changes which are frequently less clear. Notwithstanding, a significant comprehension of the sub-atomic instrument hidden medication opposition can prompt the improvement of new synthetic mixtures that can re-establish drug productivity. Witnessing the need for antimalarial drug, there is need of virtual screening of some potential drug candidates to push them up for further trials.

Catharanthus roseus is an important medicinal plant of family Apocynaceae with 70 different types of alkaloids, steroids and chemotherapeutic agents which are effective as anticancerous, antimalarial and antimicrobial activity. Vinblastine and Vincristine are two important alkaloids that are being used widely to treat diabetes.²³ Catharanthus roseus (L.) G. Don contains terpenes and alkaloids which exhibits great pharmacological activities. ²⁴ It has wide range of medicinal properties such as antioxidant, ^25-27 anticancer, ²⁸ antidiabetic, ²⁹ antimicrobial,³⁰ antiulcer,²³ hypertensive,^24,31 wound healing, ³¹ hypolipidemic ²⁷ and memory enhancement. ^28,29 With virtual screening the objective is to explore antimalarial effect of various alkaloid fractions of the plant.

Figure 1: Source: KEGG Malaria – Reference pathway – hsa pathogen Click here to view Figure

Table 2. Antimalarial drugs are classified on basis of its target:

Table 3: Antimalarial drugsdivided on basis of groups

The potential drugs and their mechanism of action is summarized in table 2, however, the effect and targets of various drugs is summarized in table 3. There are no medications for sporozoite form; the causal prophylaxis drugs repress erythrocytic stage. The suppressive prophylaxis acting medications smother clinical infection beginning. Primaquine is the main medication which targets hyphozoites of P.vivax and P.ovale as well as gametocytes of Plasmodium.

Following treatments are used to treat uncomplicated P. falciparum malaria (2015) Treating children and adults with uncomplicated P. falciparum malaria (except pregnant women in their first trimester) with one of the following ACTs:

artemether + lumefantrine

artesunate + amodiaquine

artesunate + mefloquine

dihydroartemisinin + piperaquine

artesunate + sulfadoxine-pyrimethamine (SP).

The current situation is confronting many difficulties in vector control and parasite elimination. One of the recent problems in malaria elimination is Plasmodium mutants. The mutation in Kelch protein resulted in resistance. To overcome such obstructions drug designing against target protein is been done to overcome future problem of resistance. As changes in the Kelch13 (PfK13) quality were distinguished as the vital sub-atomic markers and stayed to be key indicators of ART resistance. As far as drug delivery is concerned the most potential and safe target stage is invasion of Red Blood Cells. Regarding above fact and seeing the overview of Plasmodium infection in figure.1. Merozoite surface antigen 1,⁸ Plasmodium falciparum erythrocyte membrane protein 1,¹³ RIFIN, ¹⁵ Apical Membrane Antigen 1 are the most effective targets. However, AMA1 protein is chosen in this work because of its transmembrane nature and many unrevealed characters.¹³

This paper covers in silico designated drug adequacy against AMA1 protein quality and its viability to be utilized as medication in future for Malaria treatment.

Methodology

The FASTA grouping of P. falciparum for AMA 1 was recovered from the UniProt data set (https://www.uniprot.org/). The comparing UniProt ID explicitly for the P. falciparum strain utilized for this study was AMA1_PLAF8. The Protein sequence and 3D structure of PfK13 was derived from Uniprot data set.

Protein  Modelling

Protein modelled through Homology modelling method through Swiss Model. The maximum homology was 97% with searched templates, the modelled prepared by choosing best matching template and best model selected on basis of lowest RMSD value and DOPE score. The model structure was validated through Ramachandran plot.

Ligand Preparation

The small molecule library was built based on a wide range of experimental activity against P. falciparum. Molecular preparations such as the insertion of hydrogen bonds, 2D to 3D conversion, stereoisomers production, neutralisation of charged structures, or identification of most likely ionisation state at a user-defined pH, and the entire preparation were carried out using Maestro’s LigPrep application.³²

Protein preparation

Because the protein was simulated, the YASARA Energy Minimisation Server was used to perform an energy minimisation (Krieger et al. 2009)³³. This minimised protein was then used in the Protein Wizard tool for further processing. Using Maestro’s Glide application, the receptor grid with given grid coordinates of X = -20, Y = -10, and Z = 0 was produced with a box size of 36 × 36 × 36 Å. ³³

Molecular Docking

The produced ligand library was then docked with the protein grid using Glide’s XP (extra precision) docking function, yielding an XP Glide score as a consequence. To rank docking poses and determine protein–ligand binding affinities, the XP Glide scoring function is utilised. Using a “funnel-type” method, the Glide algorithm searches for the ligand’s position, orientation, and conformation in the enzyme’s active site. ³⁴ Maestro’s Pose viewer & XP-visualizer software was used to analyse the output files.

Protein FASTA sequence

QKY59679.1 AMA1MKSSNTKMQCIVKKLSLLAMPVVIAAILSLKIVPAGAAFVAFQTDPPSSRGNRRSSRGGRNQQAAGRQAQNEA EGTERAGGR SSSSKIIQQTPWTKYMIKYDIARCHGSGIYVDMGGYEAIGGKHYRMPIGKCPVMGKVINLASGADFLEPISADNPRYRGLGFPETVLK HTGALAGALTGTANNAINLSPVSAEDLRKWGYKGNPVTNCAEYANNIVPGSDTRTKYRYPFVYDGKDELCYVLYSPMQYNQGSRYC DADGSLEEGPSSLLCMKPYKSDLDAHLYYGSSRIDPKWDVNCPMSPIRDAIFGKWVSGACVALESAFEEFVNSAEECASILFENSATDID VDVDAEGYNEINELYSGLKNLQLKQIAFSLFAPMSKSAATAKLSKGVGKNWANYESNLGICRILSETPTCLIINAGSLAMTALGSPLESDA INFPCDIDTVGYVEPRTRNGENGESRFPVTTALSIKTLKCTKYVHSKYSESCGTYYYCSEEKSSYLSRLYQFLSNHSVKKAMAITAALLALIF AIYWVYRRLYTTKIRREHEDYDRLMSKYEYDDVSHAVSEPEQQLKTDAYIWGEAAARPSDITPVHLTKLN

Figure 2: Model prepared through modeller        Click here to view Figure

Figure 3: Superimposing modelled structure with template 6n87      Click here to view Figure

Table 4: XP Gscore and drug targets

Table 5: Structures of all compounds taken from PubChem Click here to view Figure

Figure 4: Showing docking pose of A. Secologanin, B. Serpentine, C. Ajmalicine, D. Vincristine, E. Tabersonine, F. Vindoline, G. Catharanthine, H. Catechol, I. Artemisinine Click here to view Figure

Figure 5: Showing protein ligand binding of A. Secologanin , B. Serpentine, C. Ajmalicine, D. Vincristine ,E. Tabersonine , F.Vindoline  G. Catharanthine, H. Catechol ,I. Artemisinine Click here to view Figure

Table 6: Details of the ligand protein interactions and participating amino acids

Results

Protein FASTA sequence downloaded from NCBI, BLASTp done with protein datasets. The maximum similarity was found to be 97% which suggests that homology modelling method can be used. Swiss Model software was used for searching best matched templates and best identical template was then used for preparing AMA1 model. 6n87 was found to be best match sequence and used as template for building model. Modeller was also used for building the model. Best model was selected among various models prepared on basis of DOPE score. The best model showed DOPE score -20753.71484 (Fig. 2). The modelled structured was validated by superimposing it with best match templates on basis of Root Mean Square Deviation (RMSD) which was found to be 1.65764 (Fig. 3). The structure was validated with Ramachandran plot, the plot statistics reveal that 82.6% of the residues of modelled structure were in favourable region and additional 15% were in allowed region. Only 0.7% of the residues were in disallowed region. More than 97% of the structure was in allowed regions which shows the modelled protein was good enough to carry out virtual screening. For virtual screening by using Glide’s XP (extra precision) docking function. The results were analysed by Maestro’s Pose viewer and XP- visualize software. YAARA energy minimization serevr was used to perform energy minimization.  Catharanthus roseus alkaloid fraction molecules were used as ligands against the modelled protein. Secaloganin showed Hydrogen bonding with chain A and chain B with Lysine 149 and Glutamine 147 (Fig.5 A). The shortest length H- bond is formed with Lysine 149 of Chain A of bond length 1.80881 Å. Serpentine showed two H-bond with Lysine 149 with varied bond lengths (Fig.5 B). Ajmalicine showing H-bond with Porline 218 and Lysine 149 of chain A, while one more H-bond is formed between Lysine 149 of chain B (Fig.5 C). Vincristine is a known anticancerous compound showing H-bond with Lysine 215 and Glutamine 147 of chain A and B (Fig. 5 D) respectively. Tabersonin showing hydrophilic interactions with Leucine 187, Phenylalanine 196, proline 190 of chain B and proline 220, proline 218 and glutamine 148 of chain A (Fig. 5 E). Vindoline showing H-bond with Lysine 149 and Glutamine 147 of chain B (Fig. 5 F). Catharanthine showing H-bond with Leucine 221 (Fig 5 G). Catechol forming H-bond with Lysine 149 of chain a and Glutamine 147 of chain B (Fig. 5 H). Artemisinine which is known drug of malaria shows one H- bond with Lysine 149 of chain A (Fig. 5 I). 

Discussion

In this study alkaloid fraction of Catharanthus roseus were taken for docking against AMA protein. An extensive survey of literature related to Catharanthus roseus was conducted up to Janauary,2022. Catharanthus roseus has been of prime importance in the traditional medicine systems and has wide therapeutic applications for many centuries. During phytochemical investigation total of 344 compounds including monoterpene indole alkaloids, bisindole alkaloids, flavonoids, phenolic acids and volatile were found active against many diseases in Catharanthus roseus.³⁵ AMA is a surface protein which is crucial for erythrocyte invasion of parasite, the AMA 1_PLAF8 structure is not revealed yet so protein modelling done through Swiss Model and structure validated by superimposing modelled protein with available templates. Catharanthus roseus is revealed for many therapeutic values but anti-malarial molecule is underinvestigation.³⁶ Catharanthus roseus leaf and flower extract is proved to inhibit parasite 66% Megha et al, 2017. However, when pharmacology of Catharanthus roseus leaves and flowers was done, majority of alkaloids fractions were revealed. The major molecules present in Catharanthus roseus leaf and flower extract are Secologanin, Vindoline, tabersonine, Vincristine,Serpentine, Ajmalicine,Catharanthine ³⁷ and a little fraction of catechol is been observed which is phenolflavanoid. Virtual screening is been done through Maestro and protein was simulated by YASARA energy minimization software. In virtual screening the best XP Gscore is shown by Secaloganine i.e. -7.815 Kcal/ mol while that of artemisinine is -3.226 Kcal/mol which is a known drug. Artemisinine are derived from extracts of weet wormwood and are well established for the treatment of malaria, including highly drug resistant dtrains. Its efficacy also extends to phyllogenetically unrelated parasitic infections such as schistosomiasis. In modelled protein Lysine 149 in chain A is participating in Hydrogen bond formation with secaloganin, Serpentine, Ajmalicine. Vincristine, Catechol and atremisinine. Chain Glutamine 147 is participating in hydrogen bond formation with Secaloganine, Vinscristine, Vindoline and catechol. Chain A proline 218 is participating in Hydrogen bond formation with Ajmalicine . The shortest Hydrogen bond if formed between Ajmalicine and proline 218 (1.8492 Ǻ), Chian A lysine 149 and Sepentine (1.82641 Ǻ) and Secaloganin and ChainA Lysine 149 (1.80881 Ǻ). Tabersonine is making Hydrophillic interactions with the protein’s Proline 190, Lutamine 147 pocket. Tabersonine is a monoterpenoid indole alkaloid with cytotoxic activity. It has a role as an antineoplastic agent and a metabolite.

Conclusion

The above data reveal that Secaloganine can be further taken explored as a drug molecule for antimalarial drug formulation, and chemoinformatics techniques can be used to future investigate the most stable form of the molecule in the future.

Acknowledgement

I would acknowledge the opportunity provided to me to learnt computational Biology techniques To Dr. Imran Siddiqui, Senior Scientist,  Central Drug Research Institute, Lucknow, Uttar Pradesh, India.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The authors do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

Informed Consent Statement

This study did not involve human participants, and therefore, informed consent was not required.

Clinical Trial Registration

This research does not involve any clinical trials.

Permission to reproduce material from other sources

Not Applicable 

References

1. Uhomoibhi P . Redoubling efforts to sustain seasonal malaria chemoprevention, Lancet Child Adolesc. Heal., 2022; 6( 3) :142–144. Doi- 10.1016/S2352-4642(22)00007-4
   CrossRef
2. W. H. Organization,THE E-2020 ELIMINATING 2019 progress report, 2020.
3. Naaz R, Anjum N, and Anjum G, is malaria an entirely preventable and treatable. 2020;8(1): 1248–1256. Doi – https://www.openacessjournal.primarydomain.in/abstract/104
4. Senosiain G,  Kana A, Singh I, Chourasia.S,  Das B, Dodoo M , Theisen D,  Peripheral merozoite surface proteins are targets of naturally acquired immunity against malaria in both India and Ghana. Infection and Immunity. 2020 ;88(4): 10-1128. Doi- https://doi.org/10.1128/iai.00778-19
   CrossRef
5. Jalei A, Chaijaroenkul A,  Bangchang K. Genetic Diversity of Plasmodium vivax Surface Ookinete Protein Pvs25 and Host Genes in Individuals Living along the Thai–Myanmar Border and Their Relationships with Parasite Density. Microbiology Research; 2024; 15(2): 693-707. Doi – https://doi.org/10.3390/microbiolres15020045
   CrossRef
6. Avril M, Bernabeu M , Benjamin M , Brazier J , and Smith D.Interaction between Endothelial Protein C Receptor and Intercellular Adhesion Molecule 1 to Mediate Binding of Plasmodium falciparum : Infected Erythrocytes to Endothelial Cells.mBio. 2016; 7 (4): 00615-16. https://doi.org/10.1128/mbio.00615-16
   CrossRef
7. Aly A , Vaughan A ,Kappe S, Malaria Parasite Development in the Mosquito and Infection of the Mammalian Host, Annu. Rev. Microbiol .2009;63( 1):195–221. Doi- https://doi.org/10.1146/annurev.micro.091208.073403
   CrossRef
8. Fofana F, Wele M, Diabate O, Cisse C, Diawara A, Toure M, Djimde A. (2025). Advances in Malaria Vaccine Development Targeting Plasmodium falciparum Surface Proteins. Annual Research & Review in Biology, 40(9), 1-17. DOI: https://doi.org/10.9734/arrb/2025/v40i92300
   CrossRef
9. Yan H, Feng J, Chen M.  Structural modelling prediction of recombinant Plasmodium falciparum K13-F446I and K13-C580Y gene by AlphaFold method and heterologous expression in Spodoptera frugiperda 9 cells. Pathogens. 2022;11(11): 1271.Doi-  https://doi.org/10.3390/pathogens11111271
   CrossRef
10. Kardam R, Jangra A, Kumar D, Kumar V, & Sharma V. Promising Molecular Targets of Plasmodium Falciparum, the Deadliest Parasite and Their Synthetic Inhibitors: A Review. ChemistrySelect. 2025; 10(17): 202405253. Doi- https://doi.org/10.1002/slct.202405253
    CrossRef
11. Mwakalinga S,Expression of a type B RIFIN in Plasmodium falciparum merozoites and gametes. 2012 ;11(1):1–12.Doi-  http://www.malariajournal.com/content/11/1/429
    CrossRef
12. Pei X,The ring-infected erythrocyte surface antigen ( RESA ) of Plasmodium falciparum stabilizes spectrin tetramers and suppresses further invasion. 2007 ; 110(3) : 1036–1042. Doi- https://doi.org/10.1182/blood-2007-02-076919
    CrossRef
13. Sun S, Segev-Zarko L,  Pintilie G,  Kim C,  Staggers Y,  Schmid R, Boothroyd M,   Cryogenic electron tomography reveals novel structures in the apical complex of Plasmodium falciparum.Mbio. 2024; 15(4): 02864-23. Doi- https://doi.org/10.1128/mbio.02864-23
    CrossRef
14. Dijk V, “A Central Role for P48 / 45 in Malaria Parasite Male Gamete Fertility,” 2001 vol. 104, pp. 153–164. DOI: 10.1016/S0092-8674(01)00199-4
    CrossRef
15. Goel S. RIFINs are adhesins implicated in severe Plasmodium falciparum malaria, Nat. Med. 2015; 21( 4): 314–317. Doi- https://www.nature.com/articles/nm.3812
    CrossRef
16. Yu D, Yang S,  Li Y,  Hu W.Co-expression network with protein–protein interaction and transcription regulation in malaria parasite Plasmodium falciparum. Gene. 2013;518(1): 7-16.Doi-  https://doi.org/10.1016/j.gene.2012.11.092
    CrossRef
17. Pan H., Lu X, Ye D, Feng Y, Wan, J,Ye, J. The molecular mechanism of thrombospondin family members in cardiovascular diseases. Frontiers in Cardiovascular Medicine. 2024 ; 11(1):  1337586. Doi- https://doi.org/10.3389/fcvm.2024.1337586
    CrossRef
18. Juillerat A,Lewit A, Guillotte M, Gangnard S, and Hessel A, “Structure of a Plasmodium falciparum PfEMP1 rosetting domain reveals a role for the N-terminal segment in heparin-mediated rosette inhibition,” 2011 vol. 108, no. 13, pp. 5243–5248. https://doi.org/10.1073/pnas.1018692108
    CrossRef
19. Connor C , Collins K. A Novel RNA Binding Domain in Tetrahymena Telomerase p65 Initiates Hierarchical Assembly of Telomerase Holoenzyme.  2006; 26( 6) :2029–2036. Doi-https://doi.org/10.1128/MCB.26.6.2029-2036.2006
    CrossRef
20. Das S. Processing of Plasmodium falciparum Merozoite Surface Protein MSP1 Activates a Spectrin-Binding Function Enabling Parasite Egress from RBCs Article Processing of Plasmodium falciparum Merozoite Surface Protein MSP1 Activates a Spectrin-Binding Function Enabling Parasite Egress from RBCs. Cell Host Microbe. 2015;18(4):433–444. Doi-https://doi.org/10.1016/j.chom.2015.09.007
    CrossRef
21. Yan  H, Feng J, Chen M. Structural modelling prediction of recombinant Plasmodium falciparum K13-F446I and K13-C580Y gene by AlphaFold method and heterologous expression in Spodoptera frugiperda 9 cells. Pathogens.2022;11(11):1271. Doi-https://doi.org/10.3390/pathogens11111271
    CrossRef
22. Adams J, Thrombospondins: Multifunctional Regulators of Cell Interactions.  Annu. Rev. Cell Dev. Biol. 2001; 17( 1): 25–51, Doi- https://doi.org/10.1146/annurev.cellbio.17.1.25
    CrossRef
23. Hira F, Islam A,Uddin M, Immunomodulatory and Pharmacological Properties of Catharanthus Roseus: A Comprehensive Review. Letters in Applied Nanobioscience. 2025;14(4):91-106.Doi- https://doi.org/10.33263/LIANBS142.091
24. Anvikar A, In vitro assessment of drug resistance in Plasmodium falciparum in five States of India.  Indian J. Med. Res. 2012;135(4):  494–499. https://journals.lww.com/ijmr/fulltext/2012/35040 /in_vitro_assessment_of_drug_resistance_in.8.aspx
25. Chanda E, Integrated vector management : a critical strategy for combating vector-borne diseases in South Sudan. Malaria journal. 2013;12(1):369-378. Doi-  http://www.malariajournal.com/content/ 12/1/369
    CrossRef
26. Rajput M, Nair V,  Chauhan A, Jawanjal H, Dange V, Evaluation of Antidiarrheal Activity of Aerial Parts of Vinca major in Experimental Animals. Middle East J Sci Res. 2011; 7(5): 784–788. https://www.researchgate.net/profile/Mithun-Rajput/publication/267228391_Evaluation_of_ Antidiarrheal_Activity_of_Aerial_Parts_of_Vinca_major_in_Experimental_Animals/links/546b61b80cf2397f7831bdd3/Evaluation-of-Antidiarrheal-Activity-of-Aerial-Parts-of-Vinca-major-in-Experimental-Animals.pdf
27. Takthsingji S and hospital G.evaluation of hypolipidemic activity of leaf juice of catharanthus roseus ( linn .) G . Donn . In guinea PIGS. Acta Poloniae. 2011; 68( 6): 927–935. https://ptfarm.pl/pub/File/Acta_Poloniae/2011/6/927.pdf
28. Mishra J,  Verma N, “A brief study on Catharanthus Roseus : A review,” 2017 vol. 2, no. 2, pp. 20–23. https://www.researchgate.net/profile/Navneet-Verma/publication/319007421_A_brief_study_ on_Catharanthus_Roseus_A_review/links/598ad641aca272435857d010/A-brief-study-on-Catharanthus-Roseus-A-review.pdf
29. Ramya S, Govindaraji V, Kannan K, Navaneetha, Jayakumararaj R. In Vitro Evaluation of Antibacterial Activity Using Crude Extracts of Catharanthus.  Ethnobot. Leafl. 2008; 12 (1) :  1067–1072. Doi- https://opensiuc.lib.siu.edu/ebl/vol2008/iss1/140/
30. Salman  M., Qadeer F, Pharmacological Actions and Therapeutic Potential of Trigonella foenum-graecum L. Fenugreek: Biology and applications.  2021;981(16): 523-537. Singapore: Springer Singapore. https://link.springer.com/chapter/10.1007/978-981-16-1197-1_22
    CrossRef
31. Nayak B, Anderson M, and Pereira P. Evaluation of wound-healing potential of Catharanthus roseus leaf extract in rats. Fitoterapia. 2007; 78(7) : 540–544. Doi- https://doi.org/10.1016/j.fitote. 2007.06.008
    CrossRef
32. Friesner R,  Extra Precision Glide:  Docking and Scoring Incorporating a Model of Hydrophobic Enclosure for Protein−Ligand Complexes. J. Med. Chem. 2006;  49( 21): 6177–6196. Doi- https://pubs.acs.org/doi/abs/10.1021/jm051256o
    CrossRef
33. Krieger  E, Joo  K, Lee J, Lee J, Raman S, Thompson J, Karplus K.  Improving physical realism, stereochemistry, and side‐chain accuracy in homology modeling: four approaches that performed well in CASP8. Proteins: Structure, Function, and Bioinformatics, 2016; 77(S9): 114-122. Doi- https://doi.org/10.1080/07391102.2018.1444510
    CrossRef
34. Kumar S, Singh B, Singh R. Catharanthus roseus (L.) G. Don: A review of its ethnobotany, phytochemistry, ethnopharmacology and toxicities. J. Ethnopharmacol. 2022;  284 (2) :1016-1026.Doi- https://doi.org/10.1016/j.jep.2021.114647
    CrossRef
35. Ravikumar S, Inbaneson J, Suganthi P, In vitro antiplasmodial activity of ethanolic extracts of South Indian medicinal plants against Plasmodium falciparum, Asian Pacific J. Trop. Dis.2012: 2(3): 180–183. Doi- https://doi.org/10.1016/j.jep.2021.114647
    CrossRef
36. Joshi  S, Huo C, Budhathoki R, Gurung A, Bhattarai S, Sharma K, Parajuli N. HPLC-ESI-HRMS/MS-Based Metabolite Profiling and Bioactivity Assessment of Catharanthus roseus. Plants.2025;  14(15): 2395-2410. https://doi.org/10.3390/plants14152395
    CrossRef