INTRODUCTION:

The date palm belongs to the 200 core species and over 2500 species that make up the Arecaceae family. There are around 14 species in the genus Phoenix, with Phoenix dactylifera L. being one of them¹. Ajwa dates are particularly believed to possess exceptional bioactivity qualities.

In this work, we examine the metabolic profiles of Ajwa dates gathered from Saudi Arabia and Tunisia, comparing them to various varieties of dates, in order to explore these claims². Ajwa, a unique kind of dates from Saudi Arabia (Phoenix dactylifera L.), provides an abundant supply of fiber, nutrition, and bioactive compounds. While earlier research has demonstrated the potential benefits of dates as phytoconstituents for treating renal and liver disorder³. The Ajwa dates have significant levels of energy, potassium, iron, polyphenols, flavonoids, and carbohydrates, according to chemical analysis⁴. This study examines the neuroprotective properties of methanolic Ajwa seed extract (MASE) on lipopolysaccharide (LPS)-induced cognitive impairments using several methods. Animal investigations were conducted with MASE at dosages of 200 and 400mg/kg, administered orally for thirty consecutive days. Neurotoxicity was induced by injecting four doses of LPS at a concentration of 250 g/kg intraperitoneally⁵.

Anti-mullerian hormone (AMH), sometimes called mullerian-inhibiting substance or mullerian inhibiting factor, is a glycoprotein dimer consisting of two 72 KDa monomers joined by disulfide bridges. Anti-migration hormone (AMH), a critical indicator of ovarian reserve and ovarian age, has therapeutic uses, including predicting the response to ovarian stimulation during in vitro fertilization (IVF)⁶. Previous research has shown a correlation between 25OH-D and ovarian reserve parameters, including anti-mullerian hormone (AMH) and FSH. Infertility is defined as the inability to conceive after attempting for a minimum of six months or one year, especially for women who do not use birth control. All methods of fertilization are included in assisted reproduction⁷. Infertility is clinically defined as the inability of a couple to conceive after one year of planned regular unprotected intercourse. This disorder affects around 15% of the population of reproductive age. Typically, a contributing male component may be identified in more than half of instances, with up to 40% of those cases solely attributed to male causes. Male factor infertility is frequently identified by abnormalities observed in semen tests, including a low sperm count, absence of sperm in the semen sample, reduced sperm motility, and poor sperm morphology. The assessment of female fertility involves measuring the amount of Anti-Mullerian Hormone (AMH). Infertility can arise from either male or female factors, or in rare cases, it may be classified as unexplained infertility.In order to address the problem of infertility, various procedures have been developed over time. These include medication, intrauterine insemination (IUI), conventional in vitro fertilization (IVF), IVF with intracytoplasmic sperm injection (ICSI), intracytoplasmic morphologically-selected sperm injection (IMSI), and ICSI with testicular or epididymal sperm aspiration or extraction (TESE)^8-11. Throughout history, infertile couples have been treated with great success, thanks to science and technology. Treatment for infertility is increasingly available in rural areas, but couples in these areas are still reluctant to seek treatment at hospitals because they don't know enough about the process. Government hospitals and IVF centers must take the initiative to provide patients with thorough information and guidance on the procedure¹².

Molecular docking is a widely recognized computer method used to estimate the energy of interactions between two molecules¹³. A molecular docking study is advance technique of structure based drug discovery and help to develop more heterocyle with promising pharmacological activity¹⁴, molecular docking is a well-established computational approach that relies on the structure of molecules to facilitate drug development. This methodology primarily utilizes techniques such as molecular dynamics, Monte Carlo simulation, and fragment-based search methods. Molecular docking studies are employed to ascertain the interaction between two molecules and identify the optimal orientation of a ligand that would create a complex with the lowest total energy¹⁵. Docking allows for the discovery of new therapeutic compounds, the prediction of interactions between ligands and targets at a molecular level, and the analysis of structure-activity correlations (SAR), all without previous knowledge of the chemical structure of other target modulators¹⁶. Drugs' bioactive locations may be predicted, and compounds that are suitable for experimentation can be chosen by using the binding affinities that are computed using molecular docking score algorithms^17,18.

MATERIALS AND METHODS:

Materials:

The instruments used consist of hardware and software. (software). The hardware is a set of personal computers capable of carrying out molecular calculations, modeling, and pharmacokinetic profile predictions of the ASUS Notebook Model X455LD with the technical specifications of an Intel® Inside CoreTM i5-4210U processor speed of 1.70–2.4 GHz, 4 GB of RAM, a 14-inch monitor, 640 GB of HDD capacity, and the Windows 10 Pro Home Basic 64-bit operating system. The software used is the Notepad++ program package, Autodock Tools, and the Discovery Studio Visualizer program used in the molecular docking process.

Data source:

In this study, target protein structure data was taken through the Protein Data Bank database (https://www.rcsb.org) with PDB ID 7L0J. This data is the result of the structure of the anti-Muellerian hormone type-2 receptor (AMH) that binds to AMHR2-ECD. In addition, 3D structural data of bioactive compounds from curma ajwa from some literature is required, and 3D structure subsistence is carried out on PubCheme: https://pubchem.ncbi.nlm.nih.gov.

Table 1: Compounds Of Structure

S. No

Compounds

Structure picture

Caffeic Acid

Ferullic Acid

Protocatechuic acid

Catechin

Gallic Acid

p-coumaric acid

Chlorogenic acid

Resorcinol acid

Quercetin

Luteolin

Rutin

Apigenin

kaempferol methylether

p hydroxybenzoic acid

Molecular Docking:

a. Preparation of Test:

Ligan downloaded from the PubCheme website (https://pubchem.ncbi.nlm.nih.gov) in the form of a three-dimensional structure optimized using the Autodock Tools program by setting the rotable bond. The structure is then saved in *pdbqt format.

b. Protein Preparations:

The 7L0J receptor was downloaded from the RSCB.PDB website (https://www.rcsb.org) in the *.Sdv/* 3D format. Water molecules are then removed from the structure. The protein is then charged with a Kollman charge after it is stored in the format *pdbqt.

c. Molecular Docking Process:

The ligans and receptors that have been prepared and charged are then opened using the autodock tools application connected to the vined autodoc. The first step is to determine the gridbox size on each receptor, and the next step is molecular docking using Vined Autodock. The visualization results can be seen using the Discovery Studio Visualizer.

RESULTS:

The result of redocking the 7L0J protein with the native LIGAN NAG at the gridbox coordinate points X = 28.577, Y = -29.203, and Z = 5.628 obtained the RMSD value = 2.0559A based on the result that the docking process performed was acceptable.

Figure 1: Validation of native ligans

Table 2. Molecular docking results on the 7L0J receptor

S. No	Ligan	Receptor 7L0J			ROTABLE BOND
S. No	Ligan	ΔG	RMSD i.b	RMSD u.b	ROTABLE BOND
1	Native Ligan NAG	-1.8	2.062	5.478	2
2	Caffeic Acid	-2.8	1.313	5.696	5
3	Ferullic Acid	-2.6	3.596	5.585	5
4	Protocatechuic acid	-2.8	1.049	4.598	4
5	Catechin	-2.1	1.555	6.737	6
6	Gallic Acid	-2.9	1.444	4.347	5
7	p-coumaric acid	-2.7	3.046	5.059	4
8	Chlorogenic acid	-2.3	1.149	2.276	11
9	Resorcinol acid	-2.6	2.488	3.225	0
10	Quercetin	-2.6	2.683	6.724	6
11	Luteolin	-3.0	3.057	6.814	5
12	Rutin	2.0	1.876	7.329	16
13	Apigenin	-2.1	-3.123	6.803	4
14	kaempferol methylether	-2.2	2.103	6.928	5
15	p hydroxybenzoic acid	-2.7	1.647	4.500	3

(a) (b)

Figure 2. Reseptor 7L0J Chain B (a) Native Ligan NAG (b)

Figure 3. Reseptor 7L0J Chain B dengan Native Ligan NAG

Figure 4. Reseptor 7L0J Chain B dengan Caffeic acid

Figure 5. Interaction Of Ligan NAG-4L0J

Figure 6. Interaction Of Ligan Caffeic acid-4L0J

DISCUSSION:

In the process of designing and discovering sensible medications, molecular docking is used to comprehend drug-biomolecular interactions. putting a molecule (ligan) at the receptor's preferential binding site to create a stable complex with the ability to be selective and efficacious depending on chemical interaction and binding affinity The anti-mullerian hormone receptor protein type used in this investigation was 7L0J, which was acquired from the Protein Data Bank with ID (PDB ID). Fifteen bioactive compounds from the curma ajwa were examined to see how they interacted with the protein, and these were utilized as the ligan molecules.

The first steps in the molecular docking process include ligand synthesis, water molecule removal, and Kollman charge addition to the target protein.

Redocking occurs between the target proteins and their corresponding natural ligands at this point. The native ligan of the redocking result and the actual native protein ligan are comparable, as shown by the validation result's value of RMSD (Root Mean Square Deviation). When R MSD values range from 2 to 5, the molecular docking validation method is considered satisfactory.

Fig 1 shows the process of redocking a native Ligan against the protein can also identify the binding site of the protein determined using the Gridbox. The result of the 7L0J protein redocks with the natively ligan NAG at the gridbox coordinate point X= 28.577 Y= -29.203 and Z= 5.628 obtained the value of rMSD = 2.0559A based on the result that the docking process is acceptable.

The docking result is the free-binding energy score (ΔG) of the test and protein ligand complex as well as the RMSD value. The bond-free energy indicates affinity between the ligand and the receptor (Pantsar & Poso, 2018). Low affinities indicate that the ligan and receptor require less energy to perform binding. Thus, the smaller the free bonding energy, the stronger and more stable the bond between the ligand and receptor (Syahputra et al., 2014). Free energy is obtained from the previously prepared protein-ligand docking results. The docking resulted in 20 poses with different free-binding energies. Out of 20 docked poses, the docking results for each protein were selected as the conformation with the best poses.

Table 1 shows the molecular docking results on the 7L0J receptor show different affinity values. The result of the data obtained shows that the NAG used as a comparator has a ΔG value of -1.8. The value is a standard value used to predict that the compounds that have a score of ±5% of the value have the same affinity value as the NAG for the 6L0J receptor. As for the compounds from the contents of Kurma Ajwa plants, which have a value of ΔG close to NAG, ferulic acid, catechin, chloronergic acids, rutin, apigenin, resorcinol acid, and kaempferol methylether, they are predicted to have almost similar activity to NAG.

On molecular docking, the ligan interaction is characterized by the formation of a bond between a ligan and its target protein. A hydrogen bond, a van der Waals bond, and a hydrophobic bond are observed as parameters to help determine the relationship between structure and activity. In addition, hydrophobic interactions also play a role in determining the stability of ligans against receptors. The formation of hydrophobic bonds minimizes the interaction of nonpolar residues with water.

Fig. 2 shows The docking results showed that there were two kinds of hydrogen bonds between the binding sites of proteins: conventional hydrogen (H-X), hydrogen carbon (CH-X), van der Waals, and hydrophobic bonds. The most appropriate test ligan interactions were found in caffeic acid compounds with the formation of 3 hydrogen bonds, 1 hydrogrn carbon bond, and 3 van der Waals bonds. Whereas in the NAG-4L0J complex redocking results as a comparison, there are 1 conventional hydrogen and 3 Van der Waals bands. Conventional hydrogen bands have a greater affinity compared to both hydrogen carbon and van de Waals binding, so the caffeic acid compound is predicted to have better affinities compared with the NAG-4L0J complex.

Fig. 3 and 4 shows the results of the analysis. There are some similar amino acid interactions between caffeic acid and native ligan both in hydrogen binding conformation and van der Waals, such as amino acids SER:104, GLU:75, ASN:66, and PRO:100, which are predicted to have the possibility of similar biological activity. However, the compound Caffeic Acid that forms more binding interactions than the native Ligan, the amino acids SER:101, GLN:73, and ALA:29 form hydrogen interactions, and the van der Waals are predictable to have better affinity to the anti-mullerian hormone receptor. Because of the increasing interaction of conventional hydrogen bonds with binding amino acids, it is predicted that it will increase its activity.

CONCLUSIONS:

The anti-mullerian hormone receptor has a free-binding energy value of -2.6 A, and the caffeic acid compound has the highest affinity value for these receptors according to molecular docking results. Additionally, the predicted binding interaction results showed that the NAT native ligan stimulates the receptor more effectively.

CONFLICT OF INTEREST:

The authors declare that they have no conflict of interest

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Received on 19.07.2024 Revised on 11.11.2024

Accepted on 18.02.2025 Published on 12.06.2025

Available online from June 14, 2025

Research J. Pharmacy and Technology. 2025;18(6):2803-2807.

DOI: 10.52711/0974-360X.2025.00401

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

In silico Study of Secondary Metabolite Compounds in Ajwa Dates (Phoenix dactylifera L) as an Anti-Mullerian Hormone Theraphy: Molecular Docking Approach

MATERIALS AND METHODS:

Materials:

Figure 1: Validation of native ligans

Table 2. Molecular docking results on the 7L0J receptor

S. No

Ligan

Receptor 7L0J

ROTABLE BOND

ΔG

RMSD i.b

RMSD u.b

1

Native Ligan NAG

-1.8

2.062