Evaluation of the Anti-Ulcer Activity of Rufigallol in Exper imental Models of Gastr ic

Ulcer ation in Rats

Synopsis submitted
to
School of Medical and Allied Sciences,
K. R. Mangalam University, Gurugram, Haryana
for
M. Pharm (Pharmacology) Dissertation

Super visor : Resear ch Scholar :

Dr . Swati Kaushik Ritik Rohilla
Enr ollment No.: 2304650012
CONTENTS

Sr . no. Title Page no.

1 Intr oduction 3

2 Rationale 4

3 Aim 4

4 Objective 4

5 Plan of wor k 5

6 Refer ence 6
1. INTRODUCTION

Peptic Ulcer
For more than a century, peptic ulcer disease has been a major cause of mortality. The
pathophysiology of peptic ulcer disease has centred on an imbalance between aggressive and
protective factors in the stomach.[1] The discovery of the link between Helicobacter
pylori infection and peptic ulcers by Marshall and Warren revolutionized the understanding of
peptic ulcer disease(PUD). This finding shifted the paradigm, making H. pylori a major focus in
the treatment and prevention of peptic ulcers, particularly for both duodenal and gastric ulcers.
However, other factors such as the widespread use of non-steroidal anti-inflammatory drugs
(NSAIDs) have also contributed significantly to the development of peptic ulcers, especially in
the elderly population.

Rufigallol
Rufigallol also known as 1,2,3,4,5,6,7- hexahydroxy-9,10- anthraquinone, is an organic
compound characterized by structural complexity and significant biological activity [2]. It can
be obtained by treating gallic acid with concentrated sulfuric acid followed by sodium
hydroxide treatment. It exhibit potent inhibitory action against P.falciparum [3]. It has the
ability to form π-π stacking interaction with heme protein, leading to binding and subsequent
inhibition of hemozoin formation, showing a characteristic elimination of carbomethoxy group
(CH3.) contributing to its antioxidant property and it can also inhibit the production of anti-
inflammatory mediator [4][5]. Its unique chemical structure makes it a promising candidate for
addressing various health conditions related to oxidative stress and microbial infection [6].

Pathophysiology
The Rufigallol is extensively known by its antraquininone property and identified as
antimalarial activity discovered till yet. Its unique ability to form π-π stacking interactions with
heme protein facilitate the elimination of carboxymethyl group (CH3.), suggesting notable
antioxidant properties, which contribute to its antiulcer activity. Rufigallol plays a key role in
modulating inflammation of cells by activating protein kinase A (PKA), thereby preventing the
translocation of 5-Lipoxygenase (5-LO) to the nuclear membrane – a critical step in leukotriene
synthesis [11]. Furthermore, it inhibit platelet activating factor (PAF), creating a protective
shield against mucosal injury [12]. Its anti-inflammatory properties effectively reduce the level
of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-1
beta (IL-1β), and interleukin-6 (IL-6) [13].

3
2. RATIONALE

Despite technological advancements, significantly challenges persist in identifying novel anti-

ulcer drugs to improve existing therapies. The conventional treatments such as NSAIDs, proton
pump inhibitor (PPI), histamine H2 receptor antagonists, and cytoprotective agents, aims to
reduce gastric acid secretion and alleviate symptoms of H. pylori infection. However, these
treatments have notable drawbacks, including reduced gastric juice production and severe
mucosal injury. Antibiotics are frequently employed to eradicate H. pylori, but their extensive
use contribute to antibiotic resistance, creating challenges for both individual patients and
broader public health [8]. . To address these issues, plant derived metabolites present a
promising alternative for managing ulcers without promoting antibiotic resistance. Additionally,
they are cost-efficient, readily available, multiple pharmaceutical agents, acting through
multitarget mechanism with fewer side effect and offer comparatively holistic health benefit .
This multifactorial nature of ulcer formation necessitates ongoing research to identify and
validate new therapeutic agents.[9]

3. AIM
Evaluation of antiulcer activity of rufigallol on peptic ulcers in rats.

4. OBJ ECTIVEs
 Toxicity study
Evaluation of toxicity study of rufigallol for effective dose range of rufigallol in rats for a
period of 14 days for determining its safety profile, lethal dose (LD50), and no observed
adverse effect level (NOAEL). The study aims to assess the dose-dependent toxicity, focusing
on physiological and biochemical changes, organ damage, and overall animal well-being.
 To develop ulcer model in experimental rats.
 To evaluate protective effect of Rufigallol in mitigation of peptic ulcer
 To Perform histopathology to assess mucosal damage and ulcer index
 To evaluate Biochemical parameters

4
5. PLAN OF WORK
 Toxicity study of r ufigallol
The goal is to assess the safety profile of rufigallol focusing on acute toxicity, the acute oral
toxicity studies were conducted in the rat model (both sexes), according to the method of
Litchfield and Wilcoxon (1949) and as described in “Guidelines for Testing of Chemicals-
Acute Oral Toxicity-Fixed Dose Procedure” (OECD 420, 2001).
rufigallol orally administered to a group of 6 rats after a 12 hr fast.
 Control group receive equal amount of water by gavage withaid of metal gastric cannula.
 Dose selected according to OECD guidelines protocol,assuming the best estimate of LD50
 dose progression was calculated according to geometric ratio of 1/2 (double the initial dose
to get next dose) i.e. (250 mg/kg/day, 500 mg/kg/day, 1000 mg/kg/day and 2000 mg/kg/day)
were selected for acute and sub-acute toxicity study.[12]
 The signs and symptoms associated dose were observed carefully after 0, 30, 60, 120, 180
and 240 min and then once a day for the subsequent 14 days to record toxic manifestations.
 Observations included changes in body mass, food and water consumption, hair loss, eyes
and mucus membrane and autonomic (salivation, lacrimation, perspiration, piloerection,
urinary incontinence and defecation) and the central nervous system (optosis, drowsiness,
gait, tremors and convulsion).
 On the 14th day both control and rufigallol treated animals were sacrificed, blood was
collected and analysed for haematological parameters such as red and white blood cell
counts, haemoglobin, haematocrit (HCT) and mean corpuscular volume (MCV).
 Biochemical parameters- such as blood glucose, blood urea, protein, creatine, cholesterol
and lipid profile were also assessed.
 Vital organs such as liver, kidney, heart and lungs were observed macroscopically.[13]

 Developing ulcer model in exper imental r ats

Male wistar rats weighing between 180-200g fasted for 24 hours with free access to water
administered a single dose of indomethacin (typically 20-30mg/kg) orally.

 Evaluation of pr otective effect of Rufigallol in mitigation of peptic ulcer

Male wistar rats 4 groups of 8 animals administered rufigallol in low and high dose for 14 days
control group,NSAID induced and standard treatment drug omeprazole group will be observed.

Observation - clinical signs, body weight, food and water intake, ulcer related symptom.
End of study - animal sacrified on 15th days
Observation - Daily monitoring for clinical signs, body weight, food and water intake.
 Assessed behavior for ulcer-related symptoms.

5
7.REFERENCES
1. Marshall BJ, Warren JR. Unidentified curved bacilli in the stomach of
patients with gastritis and peptic ulceration. Lancet 1984; 1: 1311–15.
2. National Center for Biotechnology Information (2024). PubChem Compound Summary for
CID 16637827, Rufigallol hexamethyl ether. Retrieved October 1, 2024
from https://pubchem.ncbi.nlm.nih.gov/compound/Rufigallol-hexamethyl-ether.
3. Modibbo, A. A., Tanko, M. M., Emmem, C. R., Muhammad, A., Abubakar, I., & Yakubu, M.
S. A Review Of Antiplasmodial Potentials Of Garcinia Kola. 2394-4404.
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Fractionation, ESI-MS Scan, Phytochemical Screening, and Antiplasmodial Activity of Afzelia
africana. Biochemistry Research International, 2022(1), 6895560.
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pain, and wounds. Journal of Ethnopharmacology, 302, 115881.
https://doi.org/10.1016/j.jep.2022.115881
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extraction and identification methods. Journal of chemical and pharmaceutical research, 8(4),
1423-1443. 0975-7384
7. Systematic review: the global incidence and prevalence of peptic ulcer disease
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activity and mechanism: a review of preclinical and clinical studies. Drug Design, Development
and Therapy, 193-213 https://doi.org/10.2147/DDDT.S446949
9. Kumar, S., Theis, T., Tschang, M., Nagaraj, V., & Berthiaume, F. (2021). Reactive oxygen
species and pressure ulcer formation after traumatic injury to spinal cord and
brain. Antioxidants, 10(7), 1013. https://doi.org/10.3390/antiox10071013
10. Han CT, Kim MJ, Moon SH, Jeon YR, Hwang JS, Nam C, et al. Acute and 28-day subacute
toxicity studies of hexane extracts of the roots of Lithospermum erythrorhizon in Sprague-
dawley rats. Toxicol Res. 2015;31:403–14
11. SSI Pillai, M Kandaswamy, S Subramanian - Journal of ApiProduct and …, 2010 -
ibra.org.uk Antiulcerogenic and ulcer healing effects of Indian propolis in experimental rat
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12. Izzo, A. A., Carlo, G. D., Mascolo, N., Capasso, F., & Autore, G. (1994). Antiulcer effect of
flavonoids. Role of endogenous PAF. Phytotherapy Research, 8(3), 179-181.
https://doi.org/10.1002/ptr.2650080313
13. Ruiz-Cano, D., & Arnao, M. B. (2024, December). Beneficial Effects of Nutraceuticals, Especially
Polyphenols on Canine Health. In Pets (Vol. 1, No. 3, pp. 228-254). Multidisciplinary Digital Publishing
Institute. https://doi.org/10.3390/pets1030017