In Vitro Anthelmintic Activities of Khaya anthotheca and Faidherbia albida Extracts Used in Chad by Traditional Healers for the Treatment of Helminthiasis and In Silico Study of Phytoconstituents

Background Helminthiasis is endemic in Chad and constitutes a public health problem, particularly among school-age children. The aim of this study was to evaluate the anthelmintic activity of extracts of Khaya anthotheca and Faidherbia albida used in Chad by traditional healers for the treatment of helminthiasis. Methods The anthelmintic activity was assessed against Heligmosomoides polygyrus and Caenorhabditis elegans larvae using the Worm Microtracker. Embryonated eggs, L1, L2, and L3 larvae of H. polygyrus were obtained after 24 h, 48 h, and 7 days of coproculture and L4 larvae of C. elegans culture using standard procedures. One hundred microliters of extracts at various concentrations, with albendazole and distilled water were, put in contact with 100 µL of H. polygyrus suspension (containing 50 parasites at various developmental stages) in a microplate and incubated for 20 h at 25°C in the Worm Microtracker. The same procedure was adopted for C. elegans, but with 180 µL of OP50. 19 µL of C. elegans suspension (containing 50 larvae) was put in contact with 1 µL of extract at various concentrations and incubated in the Worm Microtracker. Docking studies were carried out using the Schrodinger Maestro software's Glide module. The score function in the software was used to rank and group distinct possible adduct structures generated by molecular docking. Results The aqueous and ethanolic extracts of F. albida at a concentration of 2.5 mg/mL showed the same activity as albendazole (100 ± 0.00) on hatching. The IC50s of the aqueous extracts of the two plants (IC50: 0.6212 mg/mL and 0.71 mg/mL, respectively) were comparable on egg hatching of H. polygyrus with no significant difference (p ≥ 0.05) with respect to the ethanol extracts (IC50: 0.70 mg/mL and 0.81 mg/mL, respectively). There was no significant difference between the percentage inhibition of extracts and albendazole on the L1 larvae of H. polygyrus (p ≥ 0.05). The aqueous extracts acted more effectively than the ethanol extracts on the L1 larvae of H. polygyrus with an IC50 of 0.5588 and ∼9.858e − 005 mg/ml, respectively, for K. anthotheca and F. albida. The aqueous extracts of K. anthotheca and F. albida on L3 larvae of H. polygyrus had inhibitory percentages of 92.6 ± 0.62 and 91.37 ± 0.8 at 2.5 mg/mL which were lower than albendazole (100 ± 0.00). The aqueous extracts of K. anthotheca and F. albida on C. elegance showed IC50 of 0.2775 µg/mL and 0.5115 µg/mL, respectively, and were more effective than the ethanol extracts. Examining K. anthotheca and F. albida through the interaction with the protein receptor and its results also confirmed our assumption that the compound used has hydroxyl and carbonyl groups as well as aromatic rings and is exposed to phenolic and flavonoid groups in a more specific way, and it shows a better inhibitory effect. Conclusions This study scientifically validates the use of extracts of the two plants in the traditional treatment of helminthiasis. However, it will be necessary to evaluate the in vivo anthelmintic activity and toxicity. Examining the ADME properties of these compounds also supports the potential of these ligands to be transformed into pharmaceutical forms.

Background.Helminthiasis is endemic in Chad and constitutes a public health problem, particularly among school-age children.Te aim of this study was to evaluate the anthelmintic activity of extracts of Khaya anthotheca and Faidherbia albida used in Chad by traditional healers for the treatment of helminthiasis.Methods.Te anthelmintic activity was assessed against Heligmosomoides polygyrus and Caenorhabditis elegans larvae using the Worm Microtracker.Embryonated eggs, L1, L2, and L3 larvae of H. polygyrus were obtained after 24 h, 48 h, and 7 days of coproculture and L4 larvae of C. elegans culture using standard procedures.One hundred microliters of extracts at various concentrations, with albendazole and distilled water were, put in contact with 100 µL of H. polygyrus suspension (containing 50 parasites at various developmental stages) in a microplate and incubated for 20 h at 25 °C in the Worm Microtracker.Te same procedure was adopted for C. elegans, but with 180 µL of OP50.19 µL of C. elegans suspension (containing 50 larvae) was put in contact with 1 µL of extract at various concentrations and incubated in the Worm Microtracker.Docking studies were carried out using the Schrodinger Maestro software's Glide module.Te score function in the software was used to rank and group distinct possible adduct structures generated by molecular docking.Results.Te aqueous and ethanolic extracts of F. albida at a concentration of 2.5 mg/mL showed the same activity as albendazole (100 ± 0.00) on hatching.Te IC50s of the aqueous extracts of the two plants (IC 50 : 0.6212 mg/mL and 0.71 mg/mL, respectively) were comparable on egg hatching of H. polygyrus with no signifcant diference (p ≥ 0.05) with respect to the ethanol extracts (IC 50 : 0.70 mg/mL and 0.81 mg/mL, respectively).Tere was no signifcant diference between the percentage inhibition of extracts and albendazole on the L1 larvae of H. polygyrus (p ≥ 0.05).Te aqueous extracts acted more efectively than the ethanol extracts on the L1 larvae of H. polygyrus with an IC 50 of 0.5588 and ∼9.858e − 005 mg/ml, respectively, for K. anthotheca and F. albida.Te aqueous extracts of K. anthotheca and F. albida on L3 larvae of H. polygyrus had inhibitory percentages of 92.6 ± 0.62 and 91.37 ± 0.8 at 2.5 mg/mL which were lower than albendazole (100 ± 0.00).Te aqueous extracts of K. anthotheca and F. albida on

Introduction
Te poorest and most neglected communities are among those infected by soil-transmitted helminth infections, which are the most prevalent infections worldwide [1].Diferent species of parasitic worms are the cause of helminth infections that are spread through the soil [2].Tey are spread by eggs found in human feces, which contaminate the soil especially when sanitary conditions are improper.Children with infection have nutritional and physical defciencies.Periodic deworming to remove infected worms, health education to avoid reinfection, and improved sanitation to lower soil contamination with infectious eggs are the main components of control [3].Because the disease is neglected in most African countries, little efort is being made to eradicate these diseases.A study conducted by Samafou et al. [4] on intestinal helminthiases among school children in the Sahelian and Sudanian zones of Chad revealed an overall infection rate of 35.87% in both zones, with the most frequent helminths being Ascaris lumbricoides (16.41%),Schistosoma mansoni (14.0%), and Hymenolepis nana (6.53%).Brooker et al. [5] estimated that 32.7% of all rural Chadians are afected with ankylostomiasis.In the city of N'Djamena, Kostoingue et al. [6] observed that 57.7% of kids have intestinal parasites.According to recent research, 51% of people living in N'Djamena city are carriers of at least one intestinal parasite, and 60% of Chadian children who live in nomadic communities [7].All these high prevalences of helminths lead to one conclusion that this disease is seriously neglected in Chad.
Te frequent use of synthetic anthelmintics has resulted in nematode resistance to existing synthetic medications, which has negative efects such as limited efcacy.In addition, the lack of efective anthelmintics, inadequate quality, and the relatively expensive cost of medications prevent some families from getting access to them and receiving regular treatment [8].Tis is why there is a pressing need to identify and develop novel treatments that are easily accessible and less harmful to the general population in order to tackle these helminthiases.
Te use of medicinal plants, a major source of molecules with therapeutic properties as far as diseases are concerned due to the existence of natural chemicals, is one way to overcome this resistance issue [9].Due to the presence of phytochemical elements, medicinal plants are helpful in treating human ailments and play a signifcant part in healing [10].
Khaya anthotheca and F. albida are plants used in the center, west, and south of Chad for the treatment of helminthiasis by the local population.Studies have shown that aqueous and organic extracts from the bark of F. albida have antimicrobial activities [11].In the Republic of Korea, the work of Lee et al. [12] showed that K. anthotheca has antiplasmodial activity.In order to support the usage of K. anthotheca and F. albida in the traditional Chadian pharmacopoeia for the treatment of helminthiasis, the present research was carried out to evaluate the anthelmintic activity of these two plants.

Collection and Identifcation of Plant Materials.
Stem barks of K. anthotheca and F. albida were harvested from the Chadian central region.Te plants were cut into small pieces, transported in plastic bags, and identifed at the National Plant Identifcation Centre as K. anthotheca (4230/ HNC) and F. albida (45506/HCN).Te plant materials were dried and then grounded into powder and stored at room temperature.

Preparation of Extracts.
We used ethanol to make the extraction because during the survey the traditional practitioner told us to use fermented palm wine (ethanol) or infusion to prepare this remedy.Plant extracts were prepared according to the method described by Abdel Azizi et al. [13].In brief, for the ethanolic extract, 100 g of powder from K. anthotheca and F. albida were each introduced in 1 L of ethanol and the mixture was stirred for 72 h.Te homogenate was fltered using Whatman paper no. 3. Te obtained fltrate was dried at 45 °C in an oven to get the extract.For the aqueous extract, 100 g of powder was introduced in 1 L of boiled water at 100 °C and sealed until cooling.Te homogenate was fltered using Whatman paper.Te obtained fltrate was dried at 40 °C.

Reference Substances. Albendazole and levamisole
were used as the positive control while distilled water and DMSO were used as the negative control as described by Cédric et al. [14].

Isolation and Concentration of Embryonated Eggs of H. polygyrus bakeri. Te foatation technique described by
Cédric et al. [8] was used to concentrate fresh eggs.In brief, the parasite eggs foated to the top when two grams of fresh infected stool sample were mixed with a saturated NaCl solution.With the aid of a glass slide, the eggs were removed from the surface.Te slides were washed using distilled 2 Journal of Tropical Medicine water to collect the eggs.Te suspension of eggs was centrifuged three times at 1500 rpm for 10 min in order to rid of the salt solution from the eggs.Tese fresh eggs were incubated for 24 h at room temperature to produce embryonated eggs.

Culture and Collection of H. polygyrus bakeri Larvae.
Te technique described by Johnston et al. [15] was used to culture the larvae.Infected feces were mixed with charcoal in the ratio of 1 : 1 until a correct consistency was obtained.Te culture was placed on Petri dishes having wet flter paper on their surface and incubated at 27 °C for 48 h (L1 larvae), 96 h (L2 larvae), and 7 days for L3 larvae.Water was added to the cultures regularly to prevent the mixture from getting too dry, approximately every 1-2 days.
2.5.Caenorhabditis elegans Culture.Using the method outlined by Liu et al. [16], Caenorhabditis elegans (wild-type Bristol) was cultured.C. elegans was cultured on nematode growth media with OP50 (bacteria) as their food source.Synchronized colonies of eggs were incubated at 20 °C and hatched in S basalt solution.L1 larvae were then placed on nematode growth medium (NGM) and incubated for a further 36 h to obtain the L4 larvae.Te anthelminthic test was performed using these L4 larvae.
2.6.In Vitro Anthelminthic Activity against H. polygyrus bakeri and L4 Larvae of C. elegans.Te ovicidal and larvicidal activity was assessed using a Worm Microtracker as described by Cédric et al. [14].In a 96-microplate assay, 50 embryonated eggs were exposed to the extract at diferent concentrations ranging from 0.078 to 2.5 mg/mL to test for ovicidal activity.Te mobility of the L1 larvae was observed on the plate at 25 °C for 24 h in the Worm Microtracker.Te same procedure was used to test the larvicidal activity (with 50 larvae per well) against L1 to L3 H. polygyrus larvae and L4 C. elegans larvae, with the exception that the extract concentrations tested for L4 C. elegance larvae ranged from 6.25 to 200 µg/mL, and the plates were incubated in the Worm Microtracker at 20 °C for 18 h instead.Te setup was repeated three times for each treatment and control for both ovicidal and larvicidal activities in the same conditions.An infrared microbeam, which crosses each microtiter well and scanned more than ten times per second, was used by the Worm Microtracker to measure and record the motility of the worms in each well every thirty minutes.Te microbeam was interrupted when a worm moved by.Te anthelminthic activity was determined as follows: % inhibition � mobility activity of control − mobility activity of the test sample mobility activity of control × 100. (1)

Molecular Docking.
Docking is the term used to describe the process of predicting the conformation and orientation of a ligand in a particular binding site.In this study, tubulin alpha-1B chain and tubulin beta chain was/were selected as the main target for several anthelmintic substances, based on information from various articles on anthelmintic receptor proteins [17,18].Te structure of the C. elegans tubulin was downloaded from the Protein Data Bank portal (PDB ID: 6E88) as seen in Figures 1(a) and 1(b).A crystal structure of beta tubulin (PDB ID: 1SA0 (tubulin crystals complexed with colchicine binding site)) [20][21][22] from the PDB (https:// www.rcsb.org/pdb)was selected and edited after removing the heteroatoms.Te merging of nonpolar hydrogens generated polar hydrogens and Gasteiger charges (Figures 1(c) and 1(d)).By considering the relevant ionization states for both acidic and basic amino acid residues, hydrogen bonds associated with pH 7.4 were added, while crystallographic water molecules that lacked 3H bonds were deleted.Shivakumar et al. [23] used the OPLS_2005 force feld for the purpose of energy minimization of the crystal structure and we use the OPLS to minimize.Glide and SiteMap applications of Schrodinger suits [19,24] were used to determine the protein site, and docking was conducted with ligands of Khaya anthotheca [25,26] and Faidherbia albida [27][28][29].
Te tubulin alpha-1B chain and tubulin beta chain grid box were created using Maestro's Glide program in the receptor grid generation section.Te SiteMap app from Maestro [24] is used for foreseeing receptor active sites.A grid center of the tubulin alpha-1B chain was calculated (X: 240.06,Y: 91.29, Z: 118.29) by creating two boxes with ranges 15 × 15 × 15 and 20 × 20 × 20, which represent the active site of the tubulin alpha-1B chain.Te grid center of the tubulin beta chain was calculated (X: 117.09,Y: 89.98, Z: 6.33) by creating two boxes with ranges 20 × 20 × 20 and 30 × 30 × 30, which represent the active site of the tubulin beta chain.
Docking studies were carried out using the Glide module of Schrodinger Maestro software [19].Possible adduct structures generated by molecular docking were ranked and grouped using the software's score function [30] (Tables 1  and 2).Te three-dimensional structure of any complex can be predicted based on the binding properties of the ligand and he target.Te "protein preparation wizard" was used to preprocess the protein structure in Maestro [19].Te protein molecule's missing site was flled with hydrogen atoms and crucial bonds using modules' automated state generation and refnement phases.Receptor grid generation was carried out after the optimization process, and the docking scores were examined using diferent docked ligand conformations [31,32].
Journal of Tropical Medicine 2.8.In Silico ADME Evaluation.Te ADME (absorption, distribution, metabolism, and excretion) properties of K. anthotheca and F. albida compounds were predicted using QikProp application in Maestro software of Schrodinger suites [33].Te following were the standard parameters for this rule: (1) Molecular weight must not exceed 500 (2) Donor that forms hydrogen bond (range permitted: ≤5) (3) Acceptable range for hydrogen bond acceptor is 10 or less (4) LogP expresses high lipophilicity with an acceptable range of ≤5 (5) Range of acceptable molar refractivity is  To determine the ADME properties of these constituents, the QikProp module of Schrödinger Maestro (v12.5) was used [34].
2.9.Statistical Analysis.Te activity measurements were used to calculate the percentage inhibition using Microsoft Excel.Te half maximum inhibitory concentrations (IC 50 ) were then determined using the concentration-response curves produced by plotting the logarithm of the concentration as a function of the % inhibition using the GraphPad Prism version 8 software.Te Glide module of the Schrodinger Maestro software [19] was used to conduct the docking studies.Te software's score function was used to classify and rank various potential adduct structures produced by molecular docking.

Anthelminthic Activity against H. polygyrus bakeri.
Table 3 presents the percentage (%) inhibition of hatching, larvae motility, and half maximal inhibitory concentration (IC 50 ) of K. anthotheca and F. albida extracts.Te IC 50s of the aqueous extracts of the two plants (IC 50 : 0.6212 mg/mL and 0.71 mg/mL, respectively) were comparable on egg hatching of H. polygyrus with no signifcant diference (p ≥ 0.05) with respect to the ethanol extracts (IC 50 : 0.70 mg/ mL and 0.81 mg/mL, respectively).Te aqueous and ethanolic extracts of F. albida at a concentration of 2.5 mg/mL showed the same activity as albendazole (100 ± 0.00).Te aqueous and ethanolic extracts of K. anthotheca and F. albida showed a slightly lower inhibitory activity than the positive control (albendazole) on the L1 larvae with a percentage inhibition of 99.29 ± 0.25; 99.29 ± 0.35 (K.anthotheca); 100 ± 0.00; and 99.8 ± 0.12 (F.albida) at a concentration of 2.5 mg/mL.Tere is no signifcant  diference between the percentage inhibition of the extracts and albendazole (p < 0.05) on the L1 larvae.Aqueous extracts act more efectively than ethanolic extracts with an IC 50 of 0.5588 and ∼9.858e − 005 mg/mL, respectively, for K. anthotheca and F. albida on the L1 larvae.Moreover, at the concentration of 2.5 mg/ml, the aqueous and ethanolic extracts have an inhibition percentage of 98.50 ± 0.59 and 97.50 ± 0.41 for K. anthotheca and 99.68 ± 0.04 and 95.65 ± 1.12 for F. albida on the L2 larvae and the two plants show similar activity to albendazole.Tere is no signifcant diference between the percentage's inhibition of the extracts at the concentration of 2.5 mg/mL and albendazole for the L2 larvae.For the L2 stage larvae, aqueous extracts from both K. anthotheca (IC 50 0.237 mg/mL) and F. albida (0.072 mg/mL) were more effective than the ethanolic extracts (K.anthotheca: IC 50 0.21 mg/mL and F. albida 0.12 mg/mL).Te aqueous and ethanol extracts of K. anthotheca have a respective IC 50 of 0.02 and 0.06 mg/mL on L3 larvae which are more active than the aqueous and ethanol extracts of F. albida with a respective IC 50 of 0.11 and 0.31 mg/mL. 1 shows the percentage mortality vs concentration of extracts (mg/ ml) for L4 larvae of C. elegans.Aqueous extracts of K. anthotheca (IC 50 of 0.2775 µg/mL) and F. albida (0.5115 µg/mL) act more efectively than ethanolic extracts.At the concentration of 2.5 µg/mL, aqueous and ethanolic extracts of K. anthotheca (93.13 ± 1.87 and 90.70 ± 2.13) as well as F. albida (90.41 ± 0.07 and 72.62 ± 0.17) exhibited activity lower than levamisole (100 ± 0.00).Tis diference is signifcant (p < 0.05).

In Silico Approach and Molecular Docking Analysis of Anthelminthic.
A Glide module was used to perform molecular docking between the target protein and ligands [35,36].Te interaction of some ligands with amino acids in the target protein resulted in a signifcant docking score.Docking scores for the top 5 ligands are depicted in Tables 2, 4, 5, 6.
Molecular docking methodologies including HTVS, SP, and XP were used to screen the compounds extracted from K. anthotheca and F. albida.Te top 15% of the most stable ligands were evaluated for docking scores at each stage.Te most stable ligands were evaluated for XP docking score on their structures.
Te alpha tubulin-1B chain's docking scores and binding interactions with the top fve compounds of K. anthotheca are compared in Figure 2 and Table 2. 14,15-Didehydroruageanin A is the most active compound in K. anthotheca, due to its abundant hydroxyl functional groups, carboxylic acid groups, and aromatic rings, which enable it to have a stronger inhibitory efect on receptor proteins.
Due to its hydroxyl, ether, and carbonyl groups, anthothecanolide's composition also exhibits good inhibitory activity against asparagine, tyrosine, phenylalanine, and glutamic acid.
As with the preceding compounds that contained asparagine, serine, and threonine of the backbone, 3-Oacetylanthothecanolide is connected to the receptor through its carbonyl groups.In addition to its oxygen and hydroxyl groups, the khayalenoid D compound also binds to amino acids such as asparagine, glycine, and phenylalanine, showing its inhibitory efect.
Te ffth compound, khayalenoid C, shows the same inhibitory efect on the receptor protein due to its oxygen atoms as carbonyl groups and hydroxyl groups and binding to asparagine, tyrosine, and serine.Docking scores and binding interactions of the tubulin beta chain with the top fve compounds of K. anthotheca have been compared in Figure 3  acetate), show the greatest amount of interactions due to the interaction between the amino acid lysine with the hydroxyl groups and carbonyl groups.Most of the interactions between tubulin beta chain and 1,6-(furan-3-yl)-2,18,19,20tetrahydro-7,17,17-trimethyl-5,13,21-trioxahexacyclo [17.2.1.01,10.02,7.011,16.011,20]docosane-4,14-dioneare due to the interaction between lysine amino acid and the hydrogen of hydroxyl group.
Te interactions between the 1-O-     octadec-5-en-17-yl] 2methylpropanoate is based on carbonyl groups and hydrogen donors to amino acids of lysine and asparagine.Compared to albendazole, which has a carbonyl group and has an imine and imide pharmacophore on its structure, it shows similar interactions with the tubulin beta chain.

Journal of Tropical Medicine
Te alpha tubulin-1B chain's docking scores and binding interactions with the top fve compounds are compared in Figure 4 and Table 5.In F. albida, quercitrin has the most interactions with amino acids asparagine, phenylalanine, glutamic acid, and also isoleucine, due to its abundant hydroxyl functional groups, which makes it have a signifcant inhibitory efect on the receptor protein.Te apigenin compound shows a good inhibitory efect due to its hydroxyl groups and interaction with amino acids such as asparagine, phenylalanine, and isoleucine.
Tiliroside compound is attached to the receptor due to its abundant hydroxyl groups and aromatic ring, such as the previous compounds with asparagine, glycine, tyrosine, and glutamic acid.
Te composition of kaempferol is no exception to this rule and shows its inhibitory efect due to its favonoid backbone, hydroxyl and carbonyl groups, and binding to thyrosine, glutamine, glutamic acid, and asparagine amino acids.Te ffth compound of F. albida, oleanic acid also shows its inhibitory efect on the receptor protein due to its hydroxyl and carbonyl groups and binding to asparagine, and serine amino acids.Table 6 compares the tubulin beta chain's docking scores and binding interactions with the top fve compounds of F. albida.First, tiliroside interacts with tubulin beta chain receptors.A hydrogen donor is involved in these interactions because it provides hydrogen to the receptor's valine and alanine.Te hydroxyl groups of apigenin interact with the valine and asparagine of the receptor, as well as the frst structure.Kaempferol, a favonoid, provides hydrogen to the receptor's valine amino acid.Te interactions between betulin and receptors are caused by the hydroxyl groups in this compound.Despite lupeol's diferences, it has the same interactions with receptors since it provides hydrogen to Valine amino acids.
As a control, albendazole exhibits similar interactions with the tubulin beta chain due to imine and imide pharmacophores on its structure.

Evaluating the ADME.
Te drug-like behavior of a chemical agent is evaluated by the ADME property.Tables 2, 4, 5, 6 show that the constituents have good pharmacokinetic profles and are not demonstrated in violation of the Lipinski rule by the constituents.Also, they do not have any mutagenic or carcinogenic properties and are nonhazardous.

Discussion
Te aqueous and ethanolic extracts of K. anthotheca and F. albida showed good ovicidal activity against H. polygyrus.Aqueous extracts of K. anthotheca and F. albida were more active than ethanolic extracts.Tese observations are contrary to those observed by Wabo et al. [37] and Cédric et al.    [8] on H. polygyrus bakeri eggs.Te fact that egg hatching did not occur could be justifed by the active substances in the extract which crossed the egg membranes, reached the larvae, and penetrated their cuticle, thus causing their death.Moreover, when larvae seek to absorb water from the surrounding environment, they swell and break the membrane to escape, and when they fnd that the latter contains molecules harmful to its survival, it then prefers to stay and die in the membrane/shell.Regarding larvicidal activity, aqueous and ethanolic extracts of K. anthotheca and F. albida showed signifcant inhibitory activity with increasing concentration.Tese observations were similar to those obtained by Payne et al. [38].Tis could be explained by the active ingredients contained in the extracts blocking the receptors, thus paralyzing the larva.Lem et al. [39] mentioned that active compounds present in foods can pass through the intestinal wall of larvae and access the body's circulatory system.In addition, active compounds such as tannin can bind to the nematode cuticle, destabilize the membrane, and increase cell permeability by combining with sterols, resulting in death [8].

Journal of Tropical Medicine
Aqueous and ethanolic extracts of K. anthotheca exhibit larvicidal activity against L3 larvae of H. Polygyrus bakeri.Tese results show that both extracts of this plant are active.Tese observations were similar to those obtained by Ademola et al. [40], who found a good inhibition with the extract of K. senegalensis and those of Kolapo et al. [41] with K. grandifoliola.Tis activity is justifed by the presence of bioactive compounds such as terpenes, alkaloids, favonoids, phenols, and amthamines, which could contain anthelmintic properties.As for F. albida, the anthelmintic activity of the aqueous and ethanolic extracts was efective against L4 larvae of C. elegans.Tese observations validate those obtained by Mamat et al. [42], who also found the good activity of F. albida extracts against C. elegans.Tis activity could be justifed by the presence of the active principle contained in the plants and polyphenolic compounds which could have anthelmintic properties.
In silico studies and ADME assessments have revealed that kaempferol and apigenin exhibit docking scores comparable to albendazole against α-and β-tubulin, indicating strong interactions.Te hydroxyl and carbonyl groups, along with the favonoid backbone of these compounds, facilitate efective binding to tubulin chains.Tese fndings support the potential of Kaempferol and Apigenin as anthelmintic agents.
Several tubulin-binding herbicides exhibit activity against various protozoal parasites, and cell-based studies have shown that, similar to their efects in plants, the target in these protozoa is tubulin [43].Tubulin is an antiprotozoal drug target.Te benzimidazole class of anthelmintics, including albendazole, thiabendazole, mebendazole, and fenbendazole are broad-spectrum agents efective against gastrointestinal nematodes and, at higher concentrations, some trematodes.Tese anthelmintics function by binding to β-tubulin.β-tubulin, along with α-tubulin, polymerizes to form microtubules, which are essential structures within the cells of both nematodes and their host organisms [44].Te mechanism of action of benzimidazoles became clearer when mebendazole was observed to cause damage to the intestinal cells of Ascaris [45].It was found that this damage resulted from the loss of cytoplasmic tubules in the intestinal cells and the teguments of cestodes and nematodes [46].Tis disruption was associated with impaired transport of secretory vesicles and an inability of the intestinal cells to absorb glucose, ultimately leading to parasite starvation.In ascaris, mebendazole was found to bind to cytoplasmic proteins with molecular weights of 50 kDa and 100 kDa, corresponding to tubulin monomers and dimers.Benzimidazole anthelmintics were shown to compete for the β-tubulin-binding site with colchicine, a known inhibitor of cell division during metaphase [47].Microtubules are crucial for various intracellular functions, including the transport of cytoplasmic secretory vesicles [48].

Conclusion
Te aqueous extracts of K. anthotheca and F. albida showed more active anthelmintic activity in vitro than the ethanolic extracts on the eggs, L1, L2, and L3 of H. polygyrus bakeri and L4 larvae of C. elegans.In silico studies and ADME assessments have revealed that kaempferol and apigenin exhibit docking scores comparable to albendazole against α-and β-tubulin, indicating strong interactions.However, further in vivo anthelminthic and toxicity tests are required to validate their ethnobotanical usage by the local population.

Limitation of the Present Findings.
We were unable to identify the diferent ligands that might have been present in our extract by performing an HPLC fngerprint on the extracts due to our limited resources.

Table 1 :
Percentage mortality vs concentration of extracts (mg/mL) for L4 larvae of C. elegance.Te results are the mean ± SD of triplicate tests evaluated.a, b, c, d, e, f, and g values with the same superscript letter in the same column are not signifcant with diferent at P and Table4.

Table 2 :
Assessment of ADME and docking scores conducted for ligands of Khaya anthotheca and tubulin alpha-1B chain.

Table 4 :
Assessment of ADME and docking scores conducted for ligands of Khaya anthotheca and tubulin beta chain.