Phytochemical And Pharmacological Potential of Ixora Coccinea L. Against Alzheimer’s Disease: In Silico, In Vitro, And In Vivo Evaluation

R. Gayathri, P. Saranya, L. Gopi, Dr. V. Kalvimoorthi, Dr. K. Kaveri, Dr. S. Syed Abdul Jabbar Basha,

doi:10.5281/zenodo.17186332

Research Paper | Open Access
Volume 03 | Issue 09 | Article Id IJPS/250309262

Phytochemical And Pharmacological Potential of Ixora Coccinea L. Against Alzheimer’s Disease: In Silico, In Vitro, And In Vivo Evaluation
R. Gayathri* P. Saranya L. Gopi Dr. V. Kalvimoorthi Dr. K. Kaveri Dr. S. Syed Abdul Jabbar Basha
Aadhibhagawan College Of Pharmacy, Rantham, Thiruvannamalai, Tamilnadu.

Abstract

The present study investigates the therapeutic efficacy of the methanolic extract of Ixora coccinea L. (MEOI) in managing Alzheimer’s disease (AD) through in silico docking, behavioral assays, enzymatic studies, and histopathological evaluation. Key proteins associated with AD, including acetylcholinesterase (AChE) and glycogen synthase kinase-3? (GSK-3?), were analyzed for binding affinity using selected phytochemicals. Among the tested compounds, quercetin exhibited the highest binding affinity to AChE, while donepezil showed superior binding to both targets. In vivo experiments demonstrated MEOI's significant effects on cognitive function in behavioral models such as the Y-maze, Morri’s water maze, and buried food test. Additionally, MEOI improved antioxidant enzyme levels (SOD, CAT, GPx) and reduced AChE activity, with histopathological analysis confirming neuronal preservation. These findings suggest MEOI as a promising candidate for Alzheimer’s therapy through cholinergic and antioxidant mechanisms.

Keywords

Alzheimer’s disease, Ixora coccinea L., Acetylcholinesterase (AChE), GSK-3?, Antioxidants, Molecular docking, Neuroprotection

Introduction

Neurodegenerative diseases are a group of serious conditions that involve the gradual breakdown and loss of nerve cells (neurons) in the brain and spinal cord. These disorders affect movement, senses, and thinking abilities. Common examples include Alzheimer’s, Parkinson’s, ALS, Huntington’s disease, and multiple sclerosis. While each disease has different symptoms and causes, they all lead to permanent nerve damage and a steady decline in a person’s quality of life. As the global population ages, these diseases are becoming more common, creating major challenges for healthcare systems and society. Although the exact causes are still not fully understood, more research and better treatments are urgently needed. Scientists, doctors, and healthcare workers are working to understand the root causes, risk factors, and treatment options for these illnesses. Alzheimer’s disease is a serious neurological condition and the most common cause of dementia worldwide. As people live longer, the number of individuals affected continues to grow, placing a heavy burden on families, caregivers, healthcare systems, and society at large. This disease causes a gradual decline in brain function, starting with memory loss and eventually affecting a person's ability to perform daily tasks. Common symptoms include confusion, difficulty with language, changes in behavior, and trouble recognizing familiar people or places. These symptoms often begin mildly but worsen over time. The real impact of Alzheimer’s goes beyond the individual—it affects loved ones who provide care, strains healthcare services, and has major social and economic costs. The exact cause of Alzheimer’s disease is still not fully understood. However, researchers believe it involves a combination of genetic, environmental, and lifestyle factors. One of the key features of the disease is the abnormal buildup of proteins (such as beta-amyloid plaques and tau tangles) in the brain. This buildup damages and kills brain cells, leading to the progressive loss of cognitive functions.

Fig: 1 Alzheimer’s Disease

Plant Profile:

2.1 Ixora Coccinea L:

Ixora coccinea L., commonly known as jungle flame or flame of the woods, is a species of flowering plant in the family Rubiaceae. This plant is native to tropical Asia, including parts of India, Sri Lanka, and Southeast Asia, but it has since become widely cultivated in many tropical and subtropical regions around the world due to its attractive flowers.

Fig: 2 Ixora coccinea L

2.2 Taxonomical Classification:

Fig: 3 Taxonomical Classification

2.3 Chemical Constituents:

Different parts of the plant (roots, leaves, flowers, and stem) contain a variety of bioactive compounds. Major chemical groups include:

Table: 1 Chemical Constituents of Ixora coccinea L.

Plant Part	Chemical Constituents
Leaves	Flavonoids, tannins, saponins, alkaloids, glycosides, phenolic compounds
Roots	Anthraquinones, sterols, tannins, alkaloids
Flowers	Flavonoids (especially quercetin), anthocyanins, carotenoids, phenolic acids
Stem bark	Terpenoids, lignins, tannins

2.4 Medicinal Uses:

Table: 2 Traditional Medicinal Uses

Condition	Plant Part Used	Preparation / Application
Diarrhea & Dysentery	Roots	Decoction of roots
Wounds & Ulcers	Leaves, flowers	Crushed leaves or flower paste applied topically
Fever	Whole plant	Decoction or infusion
Skin diseases	Flowers, leaves	Paste or poultice for eczema, sores
Menstrual disorders	Roots	Decoction in small doses
Cough & Cold	Flowers	Used in herbal teas or syrups
Gastrointestinal disorders	Root & flower extracts	Decoctions used to relieve pain and inflammation

2.5 Pharmacological Activities (Based on Research):

Table: 3 Pharmacological Activities

Activity	Evidence (Extract / Compound)
Antimicrobial	Methanolic and ethanolic extracts effective against E. coli, S. aureus, Candida albicans
Antioxidant	High flavonoid and phenolic content in flowers and leaves
Anti-inflammatory	Leaf and root extracts reduce inflammation in animal models
Wound healing	Topical use accelerates healing (possibly due to tannins and flavonoids)
Hepatoprotective	Some extracts show liver-protective effects in experimental models
Anticancer (preliminary)	Quercetin and other flavonoids show cytotoxicity against cancer cells (in vitro)

MATERIALS AND METHODS:

3.1 Collection and Authentification of Plants:

The flowers Ixora coccinea L. was collected from the natural habitat in and around Rantham, Trivannamalai district, Tamilnadu and the plant material were duly authentifed byDr. K N Sunil kumar, Research Officer and HOD Department of Pharamacognosy , Siddha central research institute, Arumbakkam, Chennai – 600106.

Form No. PCOG002-ACF

Code. I29042521C

Part. Flower

Date. 17.06.2025

3.2 EXTRACTION:

Sample Preparation and Methanolic Extraction Of Ixora coccinea L.:The collected Flower of Ixora coccinea L. were shade dried, ground into coarse powder Individually. This fine powder (50 g) of plant material were extracted with mixture of 500 ml of Methanol 50% in Soxhlet extractor. Ixora coccinea L. (1 g) was extracted with 10 mL of 50 % Methanol at 45 °C for 8 h. The solvent was completely removed by rotary vacuum evaporator. The percentage yield of the extract will also be calculated.

Method Of Extraction: The extraction was carried out in Soxhlet apparatus. The coarse powder weighing (50g) was extracted with 500 ml of Methanol in heating mantle at 45 °C temperature for 8 hours. The combined filtrates were then evaporated under reduced pressure to give light brown viscous mass. The extract was stored at 0-4°C.

3.3 Phytochemical Analysis:

Shade dried powdered plant materials of the Flower of Ixora coccinea L. was used for the determination of the physio chemical constants in accordance with the WHO guidelines.

3.4 Preliminary Phytochemical Test:

Methanolic extract of Flower of Ixora coccinea L. was subjected to qualitative phytochemical tests to determine the presence of various phytoconstituents.

3.5 Experimental Studies:

Table: 4 Experimental Studies

Group – I	Control: Untreated
Group – II	Negative group: Scopalamine hydrobromide (1mg/kg, i.p) + vehicle (oral administration)
Group – III	Standard Donepezil (5mg/kg, orally) followed by injection of scopolamine (1mg/kg, i.p)
Group – IV	Methanolic extract of Ixora coccinea L. in Low dose) (oral administration)
Group – V	Methanolic extract of Ixora coccinea L. in High dose) (oral administration)

3.6 Pharmacological Activity:

Y Maze Apparatus
Morris Water Maze
Estimation Of Brain Cholinesterase
Assesment Of Sensation
Estimation Of Brain Acetyl Cholinesterase Level
Estimation Of Glutathione Peroxidase
Assessment Of Serum Superoxide Dimutase
Assesment Of Serum Catalase
Histopathology

3.7 Molecular Docking Study:

3.7.1 Protein:

Acetylcholinesterase (AChE) P22303 (Human AChE):

Acetylcholinesterase is a serine hydrolase enzyme that catalyzes the breakdown of acetylcholine (ACh) into acetate and choline in the synaptic cleft, thus terminating neurotransmission at cholinergic synapses.

Glycogen Synthase Kinase-3 Beta (GSK-3β) P49841

Here’s a comprehensive protein profile for Glycogen Synthase Kinase-3 Beta (GSK-3β) — UniProt ID P49841 — a key target in Alzheimer's disease research and other neurodegenerative disorders.

Fig: 4 Protein of Acetylcholinesterase (AChE) P22303 (Human AChE)

Fig: 5 Protein of Glycogen Synthase Kinase-3 Beta (GSK-3β)

3.7.2 Ligands Used for Docking:

Table: 5 Ligands Used in Docking

S. No

Compound

Structure

IUPAC Name

Mol. formula

(Mol. Wt.)

Quercetin

2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one

C₁₅H₁₀O₇

(302.24)

Gallic acid

3,4,5-Trihydroxybenzoic acid

C?H?O?

(170.12)

Ellagitannins

Polyphenolic structure

C??H??O??

(952.65)

Lupeol

1R,3aR,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-1-(2-hydroxypropan-2-yl)-3a,5a,5b,8,8,11a-hexamethyl-1,2,3,4,5,6,7,7a,11,11b,12,13,13a,13b-tetradecahydrocyclopenta[a]chrysen-9-ol

C??H??O

(426.72)

β-sitosterol

(3β)-Stigmast-5-en-3-ol

C??H??O

(414.71)

Kaempferol

3,4′,5,7-Tetrahydroxyflavone

C??H??O?

(286.24)

Syringic acid

4-Hydroxy-3,5-dimethoxybenzoic acid

C?H??O?

(198.17)

Ursolic acid

(1R,2R,4aS,6aS,6bR,8aR,10R,11R,12aR,14bR)-2,6a,6b,9,9,12a-Hexamethyl-10-hydroxy-1,2,3,4,4a,5,6,7,8,8a,10,11,12,13,14,14a,14b-octadecahydropicene-4a-carboxylic acid

C??H??O?

(456.70)

RESULTS AND DISCUSSION:

4.1 Extraction:

Table: 6 Extract Properties

S. No	Content	*Ixora coccinea L.*
1	Solvent	Methanol
2	Colour	Light Brown
3	Physical Nature	Semi liquids
4	Yield w/w	11.35

Fig: 6 Methanolic Extract of Ixora coccinea L.

4.2 Preliminary Phytochemical Analysis:

Table: 7 Preliminary Phytochemical Analysis

S. No	Physio-Chemical Constant	*Ixora Coccinea L.*	Limits (%W/W)
1	Total Ash	7.7±1.3	Not more than 10
2	Acid Insoluble Ash	1.2±1.3	Not more than 3
3	Water Soluble Extractive	17.5±1.4	Not less than 20
4	Loss On Drying	6.5	Not more than 12

4.3 Phytochemical Screening:

Fig: 7 Phyto – Chemical Test

Table: 8 Phyto – Chemical Test

S. No	Constituents	Results
1	Flavonoids	+
2	Terpenoids	+
3	Phenolic Compounds	+
4	Saponins	-
5	Tannins	+
6	Glycosides	-
7	Carbohydrates	+
8	Sterols	+
9	Alkaloids	-
10	Protein	-
11	Amino acid	-

+ = Present - = Absent

4.4 Pharmacological Activity:

4.4.1 Effect of MEOI On Y- Maze & Morris Water Maze:

Table: 9 Effect of MEOI On Y- Maze & Morris Water Maze

		Effect Of MEOI On Y- Maze			Effect Of MEOI On Morris Water Maze
S.No	Groups	% Alterations In Mice			Escape Latency (Seconds)
S.No	Groups	Day-07	Day-14	Day-21	Day-07	Day-14	Day-21
1	Control	43.34±0.44	44.22±0.35	45.49±0.44	17.11±0.35	17.22±0.36	17.66±0.31
2	Negative Control	21.22±0.35	22.24±0.36	16.11±0.46	45.12±0.36	48.66±0.44	53.77±0.56
3	Standard	22.22±0.35	25.31±0.35	27.21±0.35	46.13±0.44	45.13±0.55	36.66±0.45
4	Low Dose (MEOI -200 mg/kg )	25.43±0.35	28.20±0.35	31.06±0.35	46.11±0.39	46.19±0.35	44.65±0.35
5	High Dose (MEOI -400 mg/kg )	35.11±0.36	37.40±0.50	42.41±0.50	43.21±0.66	44.31±0.36	38.23±0.71

Fig: 8 Effect of MEOI On Y- Maze & Morris Water Maze

4.4.2 Effect of MEOI On Buried Food Test & Acetylcholinesterase:

Table: 10 Effect of MEOI On Buried Food Test & Acetylcholinesterase

		Effect Of MEOI On Buried Food Test			Effect Of MEOI On Acetylcholinesterase
S.No	Groups	Latency (seconds)			AChE Level Mmoles / Mg Protein
S.No	Groups	Day-07	Day-14	Day-21	AChE Level Mmoles / Mg Protein
1	Control	163.56±6.35	163.59±7.12	163.88±9.35	18±0.31
2	Negative Control	389.66±13.22	399.19±11.03	429.44±11.13	31.30±0.36
3	Standard	339.89±8.13	335.60±9.05	318.33±7.65	27.77±0.35
4	Low Dose (MEOI -200 mg/kg )	349.19±7.35	349.40±6.31	321.34±6.19	18.5±0.31
5	High Dose (MEOI -400 mg/kg )	345.36±7.36	341.42±7.22	326.33±5.77	21.50±0.33

Fig: 9 Effect of MEOI On Buried Food Test & Acetylcholinesterase

4.4.3 Effect of MEOI On Glutathione Peroxidase, Superoxide Dimutase & CAT:

Table: 11 Effect Of MEOI On Glutathione Peroxidase, Superoxide Dimutase & CAT

S. No	Groups	Glutathione peroxidase Level units/mg Protein	SOD (IU/ml)	CAT (units / mg protein)
1	Control	0.13±0.03	5.055 ± 0.1498	0.2553 ± 0.01046
2	Negative Control	0.21±0.03	0.8300 ± 0.1043	0.05876 ± 0.00212
3	Standard	0.07±0.03	3.490 ± 0.0631	0.2024 ± 0.008631
4	Low Dose (MEOI -200 mg/kg )	0.13±0.03	2.321 ± 0.0432	0.1535 ± 0.005765
5	High Dose (MEOI -400 mg/kg )	0.12±0.06	3.109 ± 0.0098	0.1985 ± 0.003635

Fig: 10 Effect of MEOI On Glutathione Peroxidase, Superoxide Dimutase & CAT

4.4.4 Histopathology:

Fig: 11 Histopathology Study

4.5 Molecular Docking Study:

4.5.1 Docking Data:

Table: 12 Docking Data For ACHE

S. No	Compound Code	ACHE
S. No	Compound Code	Binding Energy (kJ mol^-1)	Mode	RMSD lower bound	RMSD upper bound
	Quercetin	-8.3	2	9.087	11.22
	Gallic acid	-6.2	1	4.204	6.077
	Ellagitannins	-5.0	0	0.0	0.0
	Lupeol	-4.2	1	1.99	2.0904
	β-sitosterol	-4.93	2	1.1445	2.3357
	Kaempferol	-4.82	1	1.4902	1.5087
	Syringic acid	-4.57	1	2.8295	1.4464
	Ursolic acid	-3.55	2	0.7331	1.2851
	Donepezil	-9.17	2	8.012	6.125

Table: 13 Docking Data For GSK-3β

S. No	Compound Code	GSK-3β
S. No	Compound Code	Binding Energy (kJ mol^-1)	Mode	RMSD lower bound	RMSD upper bound
	Quercetin	–1.98	2	9.09	11.22
	Gallic acid	–1.48	1	4.20	6.08
	Ellagitannins	–1.19	0	0.00	0.00
	Lupeol	–1.00	1	1.99	2.09
	β-sitosterol	–1.18	2	1.14	2.34
	Kaempferol	–1.15	1	1.49	1.51
	Syringic acid	–1.09	1	1.45	2.83
	Ursolic acid	–0.85	2	0.73	1.29
	Donepezil	–2.19	2	6.13	8.01

Fig: 12 Compound Docked At The Receptor Of Acetylcholinesterase (AChE) (Human AChE)

Fig: 13 Compound Docked At The Receptor Of Glycogen Synthase Kinase-3 Beta (GSK-3β)

CONCLUSION:

The findings of this study highlight the therapeutic potential of the methanolic extract of Ixora coccinea L. (MEOI) in managing Alzheimer’s disease. Through molecular docking, key phytocompounds such as quercetin and gallic acid demonstrated favorable interactions with Alzheimer’s-related enzymes, particularly Acetylcholinesterase and GSK-3β, suggesting their role in modulating neurodegenerative pathways. In vivo studies reinforced these results, with MEOI exhibiting significant improvements in cognitive performance, as evidenced by behavioral models like the Y-maze, Morris water maze, and buried food test. Biochemical assessments showed that MEOI effectively lowered AChE levels while enhancing antioxidant enzyme activities (SOD, CAT, GPx), thereby mitigating oxidative stress—a central factor in Alzheimer’s pathology. Histopathological analysis further confirmed neuroprotection through preservation of brain tissue and neuronal density. Collectively, these results support the use of MEOI as a promising natural agent for cognitive enhancement and neuroprotection in Alzheimer's disease, warranting further clinical investigation and pharmacological development. The study confirms the neuroprotective efficacy of Ixora coccinea L. methanolic extract in mitigating Alzheimer’s-related pathology. By inhibiting AChE and GSK-3β, enhancing antioxidant defenses, and improving cognitive performance in vivo, MEOI demonstrates strong potential as a natural therapeutic agent. Further clinical studies and isolation of active constituents are warranted to develop phytomedicine-based strategies for AD management.

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