Therapeutic Potential of a Novel Pyridazinone Derivative in Modulating Lipid and Carbohydrate Metabolism

Reetesh Kumar Rai¹, Sudhindra Prathap², Arbind Kumar Chaudhary³, Azfar Mateen⁴,

¹Associate Professor, Department of Pharmacology, MRA Medical College, Ambedkar Nagar,

Uttar Pradesh, India.

²Assistant Professor, Department of Pharmacology, SDM College of Medical Sciences and Hospital,

Dharwad, Karnataka, India.

³Assistant Professor, Department of Pharmacology, Government Erode Medical College, Erode,

Tamil Nadu, India.

⁴Associate Professor, Department of Forensic Medicine, MRA Medical College, Ambedkar Nagar,

Uttar Pradesh, India.

*Corresponding Author E-mail: arbindkch@gmail.com

ABSTRACT:

Persistent elevation of blood glucose resulting from inadequate insulin secretion, impaired insulin action, or both, defines diabetes mellitus (DM), a chronic disorder of metabolism. Current oral hypoglycemic drugs have safety limitations and incomplete efficacy, highlighting the need for novel agents with multimodal actions. Pyridazinone derivatives possess cardioprotective, antioxidant, and anti-inflammatory activities, but their antidiabetic potential remains underexplored. This study evaluated a pyridazinone derivative in alloxan-induced diabetic rats. Animals were divided into five groups: normal control, diabetic control, glipizide (10 mg/kg), and pyridazinone (30 or 60 mg/kg) administered orally for 28 days. Key assessments included fasting glucose, serum insulin, HOMA indices, lipid profile, hepatic glycogen, enzyme activities, oxidative stress markers (MDA, SOD, CAT, GSH), cytokines (TNF-α, IL-6, CRP), liver/kidney function, and histopathology. Pyridazinone significantly reduced hyperglycemia, improved insulin sensitivity, corrected dyslipidemia, restored hepatic metabolism, and enhanced antioxidant and anti-inflammatory defenses without organ toxicity. Histology confirmed protection of liver and pancreas. These findings support pyridazinone as a promising antidiabetic candidate.

KEYWORDS: Pyridazinone, Diabetes Mellitus, Insulin Sensitivity, Oxidative Stress, Histopathology.

INTRODUCTION:

Persistent hyperglycemia brought on by either poor insulin secretion, poor insulin action, or both is a hallmark of diabetes mellitus (DM), a chronic metabolic disease. Over 537 million adults worldwide suffered from diabetes in 2021; by 2045, that number is expected to increase to 783 million¹.

In India, the prevalence has already exceeded 77 million cases, placing it among the countries with the highest disease burden². DM significantly raises the risk of cardiovascular disease, nephropathy, neuropathy, retinopathy, and non-alcoholic fatty liver disease in addition to its direct metabolic consequences. This increases morbidity and death³. Although several classes of oral hypoglycemic agents are available, their long-term use is constrained by side effects, treatment failure, and inability to provide comprehensive protection against complications⁴. The incidence and course of diabetes are significantly influenced by oxidative stress and persistent low-grade inflammation, according to pathophysiological data. Long-term hyperglycemia increases the production of excess reactive oxygen species (ROS), which causes DNA damage, protein oxidation, and lipid peroxidation⁵. These processes compromise pancreatic β-cell survival and exacerbate peripheral insulin resistance⁶. Furthermore, pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and C-reactive protein (CRP) further impair insulin signaling and enhance vascular injury⁷^,⁸. An effective antidiabetic agent, therefore, should ideally combine glucose-lowering action with antioxidant, anti-inflammatory, and organ-protective properties.

Among heterocyclic compounds, pyridazinone derivatives have gained attention for their diverse pharmacological activities, including cardioprotective, antihypertensive, anti-inflammatory, and antioxidant effects⁹^,¹⁰. Recent medicinal chemistry studies have identified structural modifications of the pyridazinone nucleus as promising leads in metabolic and inflammatory disorders. However, there is limited preclinical evidence exploring their efficacy in experimental diabetes.

The present study was undertaken to evaluate the antidiabetic, antioxidant, anti-inflammatory, and organ-protective potential of a pyridazinone derivative in alloxan-induced diabetic rats. A comprehensive analysis was performed, encompassing glycemic regulation, lipid metabolism, hepatic glycogen and enzyme activities, oxidative stress and antioxidant status, inflammatory cytokines, and safety indices, supported by histopathological evaluation of the liver and pancreas.

MATERIALS AND METHODS:

Chemicals and Drugs:

Alloxan monohydrate and glipizide and the diagnostic reagent kits of glucose, lipid and hepatic-renal function estimation were purchased under HiMedia Laboratories (Mumbai, India) and Sigma-Aldrich (St. Louis, USA). The ELISA insulin, TNF-alpha, IL-6, and CRP rat-specific ELISA kits were bought at Bioassay Technology Laboratory(Shanghai, China). They were of analytical grade and fresh solutions were prepared as needed. We have synthesized, and characterized the pyridazinone derivative used in this study, in-house in the Department of Pharmacology, MRA Medical College (Ambedkar Nagar, UP), in partnership with Kirupananda Variyar Medical College (VMKVMC), Salem, Tamil Nadu.

Animals and Ethical Approval:

Adult healthy male Wistar rats (180220g) were kept in controlled environmental conditions (temperature: 22 2C; humidity: 55 5; light/dark cycle: 12h/12h). Standard pellet feed and water ad lib were given to animals. It was found that the Institutional Animal Ethics Committee(IAEC/REETESH KUMAR RAIVU/PhD/PRMOCI7B2/KMCP/106/2020-2) was used to screen the experimental design before it was conducted in line with the CPCSEA guidelines on the care and use of laboratory animals.

Induction of Diabetes:

The onset of diabetes was induced by a one time intraperitoneal inoculation of alloxan monohydrate (150 mg/kg) freshly dissolved in normal saline. To avoid early hypoglycemia after indiciting, the animals were fed with 5% glucose solution in 24hours. In 72hours a determination of fasting blood glucose (FBG) was done and rats with FBG reading above 250mg/dL were defined as diabetic and were represented in the study.

Experimental Design:

The study comprised five groups of Wistar rats, with five animals in each group (n = 5). Group I served as the normal control and received the vehicle only. Group II represented the diabetic control, in which diabetes was induced using alloxan but no treatment was administered. Group III received glipizide at a dose of 10mg/kg orally, serving as the standard reference. Group IV and Group V were administered pyridazinone at doses of 30mg/kg and 60mg/kg orally, respectively. All treatments were administered once daily by oral route for a period of 28 consecutive days. To monitor changes in physical condition and treatment response, body weights were recorded on days 0, 14, and 28 throughout the experimental duration.

Biochemical Analysis and Sample Collection:

At the end of the treatment period, blood samples were collected from each rat by retro-orbital puncture under light anesthesia. The separated serum samples were stored at −20°C until further use for biochemical and cytokine evaluations. Following blood collection, the liver and pancreatic tissues were carefully excised, rinsed with ice-cold saline, and preserved appropriately for subsequent histopathological examinations.

The biochemical investigations were conducted using established standard procedures. Glycemic indices were assessed by measuring fasting blood glucose (FBG) through the glucose oxidase–peroxidase method¹¹, serum insulin levels (ELISA), and calculating HOMA-IR and HOMA-β indices¹². The lipid profile included determination of total cholesterol (TC)¹³, triglycerides (TG)¹⁴, high-density lipoprotein (HDL)¹⁵, low-density lipoprotein (LDL)¹⁶, and phospholipids¹⁷, along with derived ratios such as TC/HDL, LDL/HDL, and the Atherogenic Index of Plasma (AIP). Parameters related to hepatic metabolism were evaluated by estimating hepatic glycogen¹⁸, hexokinase and glucokinase¹⁹, and glucose-6-phosphatase²⁰ activity. To assess oxidative stress and antioxidant defense, levels of malondialdehyde (MDA)²¹, superoxide dismutase (SOD)²², catalase (CAT)²³, and reduced glutathione (GSH)²⁴ were measured. Inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and C-reactive protein (CRP), were quantified by ELISA, and the Inflammatory Load Index (ILI) was computed relative to the diabetic control group. Finally, liver and kidney function markers—alanine aminotransferase (ALT), aspartate aminotransferase (AST)²⁵, urea²⁶, and creatinine²⁷—were analyzed, and the corresponding AST/ALT and urea/creatinine ratios were calculated to assess hepatic and renal integrity.

Histopathological Evaluation:

Samples of the liver and pancreas were fixed on 10% neuter-buffered formalin, processed, and embedded in paraffin. Staining with hematoxylin and eosin (H&E) was done in sections of 5 µm thickness. Microscopic analysis was done with a scale bar of 50 μm at 40 x magnification with attention paid to the hepatocellular integrity, fatty degeneration, inflammatory infiltration, and architecture of the islets in the pancreas. Photographs of the representatives were recorded to be analyzed.

Statistical Analysis:

The data were given in mean ± SEM (n = 5). The one-way ANOVA with a post-hoc test using SPSS version 25 was used to test the statistical differences between groups. The significant level was accepted at P < 0.05. Tables and figures containing superscripts refer to:

• a = vs Normal Control

• b = vs Diabetic Control

RESULT:

Glycemic Regulation and Insulin Sensitivity:

Persistent hyperglycemia was observed in the diabetic control group throughout the experimental period, with concomitant hypoinsulinemia and abnormal insulin indices. In contrast, pyridazinone treatment resulted in significant dose-dependent improvements in glycemic control and insulin sensitivity, approaching the efficacy of glipizide.

Diabetic rats exhibited severe hyperglycemia with markedly reduced insulin levels, reflected by increased HOMA-IR and reduced HOMA-β, confirming insulin resistance and β-cell dysfunction. Pyridazinone treatment significantly lowered fasting glucose, restored serum insulin, reduced HOMA-IR, and improved HOMA-β in a dose-dependent manner. Although glipizide produced the greatest effect, pyridazinone (especially at 60mg/kg) demonstrated substantial efficacy, suggesting dual action on insulin sensitivity and β-cell preservation.

Lipid Profile:

Diabetic rats exhibited a typical dyslipidemic pattern, with approximately a two-fold rise in total cholesterol and triglycerides, a 45% increase in LDL and phospholipids, and a 40% reduction in HDL compared with normal controls. These changes mirror human diabetic dyslipidemia reported in clinical studies, where LDL/HDL ratio and TC/HDL ratio are strong predictors of cardiovascular morbidity. Administration of pyridazinone significantly corrected these abnormalities in a dose-dependent manner. At 60mg/kg, pyridazinone restored lipid indices closer to those of the glipizide group, particularly in HDL elevation and LDL/HDL ratio reduction.

Table 1. Effects of Pyridazinone on Fasting Glucose, Insulin Levels, and Insulin Sensitivity Indices in Alloxan-Induced Diabetic Rats (Mean ± SEM, n = 5)

Group	Baseline Glucose (mg/dL)	Mid Glucose (mg/dL)	Final Glucose (mg/dL)	Serum Insulin (µIU/mL)	HOMA-IR	HOMA-β
Normal Control	85.2 ± 2.0	90.1 ± 2.5	88.3 ± 2.3	14.2 ± 1.1	2.98 ± 0.22	112.5 ± 8.4
Diabetic Control	182.4 ± 5.1^a	195.2 ± 6.2^a	200.6 ± 6.5^a	6.8 ± 0.9^a	9.25 ± 0.74^a	32.1 ± 4.2^a
Glipizide (10 mg/kg)	184.1 ± 5.0	120.2 ± 4.0^b	110.4 ± 3.5^b	12.9 ± 1.2^b	3.62 ± 0.31^b	95.7 ± 7.5^b
Pyridazinone 30 mg/kg	180.6 ± 4.8	135.4 ± 4.8^b	125.2 ± 5.0^b	10.4 ± 1.0^b	4.92 ± 0.41^b	78.3 ± 6.2^b
Pyridazinone 60 mg/kg	178.3 ± 4.7	130.6 ± 5.0^b	120.7 ± 5.2^b	11.6 ± 1.1^b	4.25 ± 0.38^b	88.9 ± 7.1^b

Data are mean ± SEM (n = 5); ANOVA p < 0.001.

Table 2. Effects of Pyridazinone on Serum Lipid Profile and Cardiovascular Risk Indices in Alloxan-Induced Diabetic Rats (Mean± SEM, n = 5)

Group	TC (mg/dL)	TG (mg/dL)	HDL (mg/dL)	LDL (mg/dL)	Phospholipids (mg/dL)	TC/HDL	LDL/HDL	AIP (log TG/HDL)	% Change in TC vs Diabetic
Normal Control	90.8 ± 2.7	96.4 ± 2.8	50.5 ± 1.8	16.4 ± 1.5	126.5 ± 3.5	1.8	0.32	-0.22	–
Diabetic Control	205.8 ± 5.3^a	160.6 ± 4.6^a	30.7 ± 1.3^a	42.7 ± 2.8^a	226.4 ± 5.3^a	6.7	1.39	0.72	Reference
Glipizide (10 mg/kg)	120.8 ± 3.4^b	105.8 ± 3.2^b	46.2 ± 1.7^b	22.7 ± 1.9^b	150.4 ± 3.8^b	2.6	0.49	-0.04	-41%
Pyridazinone 30 mg/kg	131.5 ± 3.6^b	116.7 ± 3.4^b	38.4 ± 1.4^b	32.3 ± 2.2^b	162.6 ± 4.1^b	3.4	0.84	0.48	-36%
Pyridazinone 60 mg/kg	125.4 ± 3.6^b	109.4 ± 3.2^b	43.4 ± 1.5^b	25.3 ± 2.0^b	154.5 ± 4.0^b	2.9	0.58	0.40	-39%

Data are mean±SEM (n = 5); ANOVA showed significant lipid variations (p < 0.001).

The lipid-lowering effect of pyridazinone was evident across all parameters, with a clear dose–response relationship. The 30mg/kg dose produced moderate improvements, while 60mg/kg achieved near-normalization of lipid indices, particularly in restoring HDL and reducing LDL and atherogenic ratios. Compared with glipizide, pyridazinone demonstrated slightly lower efficacy but still produced clinically relevant reductions in cardiovascular risk markers. These findings suggest that pyridazinone not only improves glycemic control but also exerts cardioprotective benefits by ameliorating diabetic dyslipidemia.

Hepatic Glycogen and Enzyme Activities:

Alloxan-induced diabetic rats showed severe impairment in carbohydrate metabolism, reflected by a >65% reduction in liver glycogen and marked suppression of hexokinase and glucokinase activities, along with an abnormal increase in glucose-6-phosphatase activity. This enzymatic profile indicates a metabolic shift toward gluconeogenesis and impaired glycolysis. Treatment with pyridazinone significantly reversed these abnormalities in a dose-dependent manner. The higher dose (60mg/kg) restored hepatic enzyme activity more effectively than the 30 mg/kg dose, approaching the efficacy of glipizide.

Pyridazinone administration enhanced hepatic glycogen storage and improved glycolytic enzyme activities (hexokinase and glucokinase) while simultaneously reducing excessive glucose-6-phosphatase activity, thereby correcting the imbalance between glycolysis and gluconeogenesis seen in diabetic rats. The 60mg/kg dose was more effective than 30mg/kg, though glipizide produced the strongest restoration. These results demonstrate that pyridazinone improves hepatocellular glucose utilization and reduces endogenous glucose output, contributing to its overall antihyperglycemic efficacy.

Oxidative Stress and Antioxidant Status:

Diabetic rats exhibited a characteristic oxidative imbalance: lipid peroxidation was markedly increased (MDA ↑), while antioxidant defenses (SOD, CAT, GSH) were significantly suppressed. Pyridazinone treatment reduced oxidative stress and restored antioxidant capacity in a dose-dependent manner, with the 60mg/kg group nearly matching glipizide.

Pyridazinone treatment reduced lipid peroxidation (MDA ↓36–41%) and enhanced antioxidant defenses (SOD ↑55–71%, CAT ↑49–61%, GSH ↑77–100% compared with diabetic controls). The 60mg/kg dose showed greater efficacy than 30mg/kg, indicating a clear dose–response effect. While glipizide achieved the highest normalization, pyridazinone demonstrated robust antioxidative potential, strongly supporting its role in restoring redox balance and protecting tissues from oxidative injury associated with diabetes.

Inflammatory Cytokines:

Systemic inflammation was markedly increased in diabetic rats, as reflected by elevated TNF-α, IL-6, and CRP levels compared with normal controls. Treatment with pyridazinone drastically reduced these pro-inflammatory cytokines in a dose-dependent manner, with the higher dose approaching the efficacy of glipizide.

Table 4. Effects of Pyridazinone on Oxidative Stress and Antioxidant Status in Alloxan-Induced Diabetic Rats (mean ± SEM; n = 5)

Group	MDA (nmol/mg protein)	SOD (U/mg protein)	CAT (U/mg protein)	GSH (µmol/g tissue)	%Δ vs DC (MDA)	%Δ vs DC (SOD)	%Δ vs DC (CAT)	%Δ vs DC (GSH)
Normal Control	2.10 ± 0.12	9.10 ± 0.35	51.2 ± 2.1	8.20 ± 0.31	↓62.5%	↑116.7%	↑101.6%	↑164.5%
Diabetic Control	5.60 ± 0.20^a	4.20 ± 0.22^a	25.4 ± 1.4^a	3.10 ± 0.18^a	Reference	Reference	Reference	Reference
Glipizide (10 mg/kg)	3.00 ± 0.15^b	7.80 ± 0.30^b	43.6 ± 1.9^b	6.80 ± 0.27^b	↓46.4%	↑85.7%	↑71.7%	↑119.4%
Pyridazinone 30 mg/kg	3.60 ± 0.14^b	6.50 ± 0.28^b	37.8 ± 1.7^b	5.50 ± 0.23^b	↓35.7%	↑54.8%	↑48.8%	↑77.4%
Pyridazinone 60 mg/kg	3.30 ± 0.13^b	7.20 ± 0.29^b	40.9 ± 1.8^b	6.20 ± 0.25^b	↓41.1%	↑71.4%	↑61.0%	↑100.0%

Data are mean ± SEM (n = 5); ANOVA p < 0.001.

Table 5. Effects of Pyridazinone on pro-inflammatory cytokines in alloxan-induced diabetic rats (mean ± SEM; n = 5)

Group	TNF-α (pg/mL)	%Δ vs DC (TNF-α)	IL-6 (pg/mL)	%Δ vs DC (IL-6)	CRP (mg/L)	%Δ vs DC (CRP)	TNF-α/IL-6	Inflammatory Load Index
Normal Control	22.0 ± 1.8	↓60.0%	18.0 ± 1.5	↓60.0%	0.35 ± 0.03	↓75.0%	1.22	0.28
Diabetic Control	55.0 ± 2.3^a	Reference	45.0 ± 2.0^a	Reference	1.40 ± 0.08^a	Reference	1.22	1.00
Glipizide (10 mg/kg)	28.0 ± 1.9^b	↓49.1%	23.0 ± 1.6^b	↓48.9%	0.55 ± 0.04^b	↓60.7%	1.22	0.42
Pyridazinone 30 mg/kg	35.0 ± 2.0^b	↓36.4%	30.0 ± 1.8^b	↓33.3%	0.80 ± 0.05^b	↓42.9%	1.17	0.62
Pyridazinone 60 mg/kg	31.0 ± 1.9^b	↓43.6%	26.0 ± 1.7^b	↓42.2%	0.65 ± 0.04^b	↓53.6%	1.19	0.50

Data are mean ± SEM (n = 5); ANOVA p < 0.001.

Table 6. Effects of pyridazinone on liver enzymes and kidney function markers in alloxan-induced diabetic rats (mean ± SEM; n = 5)

Group	ALT (U/L)	%Δ vs DC (ALT)	%Δ vs NC (ALT)	AST (U/L)	AST/ALT	%Δ vs DC (AST)	%Δ vs NC (AST)
Normal Control (NC)	42.0 ± 2.5	—	—	75.0 ± 3.8	1.79	—	—
Diabetic Control (DC)	85.0 ± 3.6^a	Ref	↑102%	150.0 ± 5.2^a	1.76	Ref	↑100%
Glipizide (10 mg/kg)	48.0 ± 2.8^b	↓43.5%	↑14%	82.0 ± 4.0^b	1.71	↓45.3%	↑9%
Pyridazinone 30 mg/kg	52.0 ± 3.0^b	↓38.8%	↑24%	95.0 ± 4.2^b	1.83	↓36.7%	↑27%
Pyridazinone 60 mg/kg	49.0 ± 2.7^b	↓42.4%	↑17%	88.0 ± 4.1^b	1.80	↓41.3%	↑17%

Group	Urea (mg/dL)	%Δ vs DC (Urea)	%Δ vs NC (Urea)	Creatinine (mg/dL)	Urea/Creat Ratio	%Δ vs DC (Creat)	%Δ vs NC (Creat)
Normal Control (NC)	28.0 ± 1.6	—	—	0.75 ± 0.05	37.3	—	—
Diabetic Control (DC)	55.0 ± 2.8^a	Ref	↑96%	1.40 ± 0.08^a	39.3	Ref	↑87%
Glipizide (10 mg/kg)	31.0 ± 1.7^b	↓43.6%	↑11%	0.82 ± 0.06^b	37.8	↓41.4%	↑9%
Pyridazinone 30 mg/kg	36.0 ± 1.8^b	↓34.5%	↑29%	0.92 ± 0.06^b	39.1	↓34.3%	↑23%
Pyridazinone 60 mg/kg	33.0 ± 1.7^b	↓40.0%	↑18%	0.85 ± 0.05^b	38.8	↓39.3%	↑13%

Data are mean ± SEM (n = 5); ANOVA p < 0.001.

Table 7. Physiological outcomes in alloxan-induced diabetic rats (mean±SEM; n = 5)

Group	Initial BW (g)	Mid BW (g)	Final BW (g)	%Δ Final vs Initial	Food intake (g/day)	Survival (%)
Normal Control	210.4 ± 4.2	218.6 ± 4.0	225.2 ± 4.5	↑7.0%	22.5 ± 1.0	100
Diabetic Control	212.5 ± 3.8	185.7 ± 4.5^a	170.3 ± 4.7^a	↓19.9%	28.6 ± 1.2^a	100
Glipizide (10 mg/kg)	211.3 ± 3.9	207.5 ± 4.1^b	215.6 ± 4.2^b	↑2.0%	23.1 ± 1.1^b	100
Pyridazinone 30 mg/kg	212.0 ± 4.0	195.8 ± 4.3^b	200.2 ± 4.6^b	↓5.6%	25.2 ± 1.1^b	100
Pyridazinone 60 mg/kg	210.8 ± 3.7	200.6 ± 4.2^b	206.3 ± 4.3^b	↓2.1%	24.1 ± 1.0^b	100

Data are mean ± SEM (n = 5); ANOVA p < 0.001.

Figure 1 Histopathology of Liver and Pancreas

Pyridazinone significantly attenuated diabetes-induced body weight loss and moderated hyperphagia, with the 60mg/kg dose performing better than 30mg/kg. While glipizide fully normalized weight and food intake, pyridazinone demonstrated a clear protective effect on metabolic stability, further supporting its therapeutic potential in diabetes.

Histopathology of Liver and Pancreas:

Histopathological evaluation provided structural confirmation of the biochemical findings. Normal control livers (Panel A) exhibited well-preserved hepatic cords radiating from a central vein with intact hepatocyte morphology. In contrast, diabetic control livers (Panel B) showed marked architectural disruption, with microvesicular steatosis, ballooned hepatocytes, and dense inflammatory cell infiltration. Pyridazinone-treated livers (Panel C) displayed near-normal histology, characterized by reduced steatosis, restored hepatocellular architecture, and diminished inflammatory changes.

The normal pancreas (Panel D) displayed intact islets of Langerhans with abundant β-cells, whereas the diabetic pancreas (Panel E) showed shrunken, degenerated islets with marked β-cell loss. Pyridazinone-treated pancreas (Panel F) exhibited preserved islet structure, improved β-cell density, and restoration of architecture. Similarly, pyridazinone mitigated hepatic steatosis and degeneration, yielding near-normal morphology. These histological findings corroborate the biochemical evidence of glycemic control, hepatoprotection, and pancreatic preservation.

DISCUSSION:

The present study demonstrates that pyridazinone exerts significant antidiabetic, antihyperlipidemic, antioxidant, and anti-inflammatory effects in alloxan-induced diabetic rats. The compound improved glycemic regulation, corrected dyslipidemia, preserved hepatic and renal function, and mitigated pancreatic β-cell damage, thereby providing a multi-faceted protective profile. Persistent hyperglycemia and hypoinsulinemia in diabetic controls confirmed severe insulin resistance and β-cell dysfunction, consistent with previous alloxan models. Pyridazinone significantly reduced fasting glucose, improved HOMA-IR, and restored HOMA-β in a dose-dependent manner, with the 60mg/kg dose approaching the efficacy of glipizide. These findings suggest that pyridazinone enhances both peripheral insulin sensitivity and beta cell function, likely through preservation of pancreatic islets as supported by histopathology^27,28. Dyslipidemia is a hallmark of type 2 diabetes and contributes substantially to cardiovascular morbidity. Diabetic controls showed marked increases in total cholesterol, triglycerides, LDL, and phospholipids, with reduced HDL, mirroring clinical diabetic dyslipidemia. Pyridazinone corrected these abnormalities, with notable improvements in HDL restoration and LDL/HDL ratio reduction, resulting in a decreased Atherogenic Index of Plasma. These cardioprotective effects are comparable to those reported for established antidiabetic drugs, supporting pyridazinone’s translational relevance. Diabetes-induced impairment of hepatic glycolysis and enhanced gluconeogenesis was evidenced by reduced glycogen content, decreased hexokinase and glucokinase activity, and increased glucose-6-phosphatase activity. Pyridazinone restored these enzymatic activities, favoring glycolytic flux and hepatic glycogen storage, while attenuating excessive gluconeogenesis. These changes indicate improved hepatic insulin responsiveness, which contributes to systemic glucose lowering²⁹. Oxidative stress is a central pathogenic mechanism in diabetic complications. The elevated MDA levels and reduced antioxidant enzymes (SOD, CAT) and GSH in diabetic controls highlight this imbalance. Pyridazinone treatment reduced lipid peroxidation and enhanced antioxidant defenses, with the 60mg/kg dose producing equal to complete normalization. These data indicate that pyridazinone restores redox balance, potentially protecting against oxidative tissue injury associated with chronic hyperglycemia³⁰.

Low-grade systemic inflammation contributes to insulin resistance and diabetic vascular complications. Elevated TNF-α, IL-6, and CRP in diabetic controls are consistent with this inflammatory phenotype. Pyridazinone significantly reduced these cytokines, lowering the Inflammatory Load Index by ~50%. This immunomodulatory action complements its metabolic effects and underscores pyridazinone’s potential to mitigate inflammation-driven diabetic complications^31,32. Unlike many synthetic antidiabetic agents that may cause hepatotoxicity or nephrotoxicity, pyridazinone demonstrated a favorable safety profile. ALT, AST, urea, and creatinine levels were significantly reduced compared with diabetic controls, and derived indices (AST/ALT ratio, Urea/Creatinine ratio) remained within physiological ranges. These findings strongly suggest that pyridazinone is safe, organ-protective, and well tolerated³³.

Attenuation of body weight loss and normalization of food intake further confirmed pyridazinone’s systemic efficacy. Importantly, histopathology demonstrated preserved hepatic and pancreatic architecture, with restored hepatocyte cords and improved β-cell density. These structural findings provide direct evidence for pyridazinone’s protective effects at the tissue level, aligning with the biochemical results.

Table 8: Comparative Analysis (Past ~10 Years; rodent diabetes models)

Study (Year)	Model and Duration	Intervention (dose/route)	Glycemic Control	Lipids	Oxidative Stress	Inflammation	Safety / Histology	Comparator
Hamadjida et al., 2024	Alloxan Wistar rats; 21 days	Boswellia dalzielii or Hibiscus sabdariffa extracts (100–400 mg/kg, p.o.)	↓FPG; improved vs diabetic control	↓TC, TG; ↑HDL (reported previously; this study focused redox/inflammation)	↓MDA; ↑GSH, SOD, CAT (p < 0.01–0.001)	↓TNF-α, ↓IL-6, ↓IL-1β (p < 0.01–0.001)	–	Glibenclamide 10 mg/kg
Bukhari et al., 2024	Alloxan rats; 28 days	Butin 25/50 mg/kg (p.o.)	↓Glucose, ↓HbA1c; ↑Insulin (p < 0.05–0.0001)	↓TC, TG; ↑HDL	↓MDA; ↑SOD, ↑CAT, ↑GSH	↓TNF-α, ↓IL-6, ↓IL-1β; ↓NF-κB	↓ALT/AST; ↓creatinine; Pancreas histology restored	–
Guo et al., 2022 (npj Sci Food)	T2D model rats; 4 weeks	Oral SOD (native vs liposomal)	↓Blood glucose; ↑insulin sensitivity; ↑AMPK	Improved lipid panel vs model; some > metformin	Colon: ↓MDA; ↑SOD, CAT, GPx	↓LPS influx; ↓cytokines	Well tolerated; gut barrier histology improved	Metformin
Aslam et al., 2023	Alloxan rats; 6 weeks	Polyphenol-rich polyherbal mixture (200–600 mg/kg, p.o.)	↓FBG (dose-dependent)	↓TC, TG, LDL/VLDL; ↑HDL; ↓AI/CRI	↓Oxidative stress markers	↓TNF-α (hepatic/renal tissue)	Liver and kidney histology improved	Glibenclamide
Alrefaei and Elbeeh, 2025 (MDPI Biology)	STZ rats; ~4 weeks	Glycyrrhiza glabra extract	↓Glucose; ↑insulin sensitivity	Improved lipid profile	↓MDA; ↑GSH, SOD, CAT	↓Inflammation; gene-level antioxidant upregulation	↓ALT/AST/ALP; Liver histology restored	–
Abdullah et al., 2018 (Biomed Pharmacother)	Alloxan rats; 4 weeks	Niacin (Vit-B3) (10–15 mg/kg, p.o.)	↓FBG (dose-dependent)	–	↓Oxidative stress; DNA damage reduced	↓Inflammatory markers (reported)	–	–
Alharbi et al., 2022 (MDPI Antioxidants)	Diabetic rats	6-Gingerol	↓Hyperglycemia-linked renal injury	–	↑SOD/CAT/GPx; ↓MDA	↓Inflammatory cytokines; nephroprotection	↓Urea/Creatinine	–

FPG: fasting plasma glucose; AI: atherogenic index; CRI: coronary risk index. Arrows indicate direction vs diabetic control. Where exact numerical effect sizes were not available in abstracts/accessible text, direction and significance reported per source.

Across contemporary rodent models, antidiabetic interventions that succeed tend to share a triad of effects: (i) robust glycemic control; (ii) anti-oxidative actions (↓MDA; ↑SOD/CAT/GSH); and (iii) anti-inflammatory signaling (↓TNF-α/IL-6/NF-κB), frequently accompanied by hepato-renal safety and histological rescue. Plant-derived small molecules like butin reproduced the full triad—including improved lipids, lower ALT/AST, and preserved pancreatic islets—mirroring the breadth of endpoints we targeted here, and doing so in an alloxan model directly comparable to ours. Likewise, polyphenol-rich mixtures and licorice extracts not only normalized dyslipidemia but also reversed hepatic oxidative injury on microscopy, aligning with our biochemical and histopathology readouts. Complementary mechanistic evidence from oral SOD demonstrates that restoring redox balance and barrier function upstream can cascade into lower systemic cytokines and glucose, offering a plausible systems-level explanation for the concurrent metabolic and inflammatory improvements in our work.^32,33 Finally, single-entity antioxidants including galangin, niacin, and 6-gingerol consistently lower lipid peroxidation and cytokines while improving glycemia or renal outcomes—supporting the view that durable antihyperglycemic efficacy in these models is tightly coupled to redox and immune modulation, not just insulin secretagogue activity. Positioning your pyridazinone: Framed against this literature, pyridazinone’s simultaneous improvements in glycemia, lipid ratios, hepatic glycogen/enzymes, oxidative stress (MDA↓; SOD/CAT/GSH↑), inflammation (TNF-α/IL-6/CRP↓) and organ safety situate it among top-tier multi-target agents rather than narrow glucose-lowering drugs—an angle favored.

Together, these results position pyridazinone as a promising candidate for diabetes management, acting through a multimodal mechanism: improving insulin sensitivity, correcting dyslipidemia, restoring hepatic metabolism, reducing oxidative stress, and attenuating inflammation. However, the study is limited by its short duration, relatively small sample size, and use of an alloxan-induced model, which primarily mimics type 1-like β-cell injury. Further studies in high-fat diet or genetic models of type 2 diabetes, as well as chronic toxicity and pharmacokinetic evaluations, are warranted to confirm its translational potential.

CONCLUSION:

Taken together, pyridazinone’s simultaneous improvements in glycemia, lipid ratios, hepatic metabolism, oxidative stress, inflammation, and organ safety position it as a promising candidate for diabetes management. Its multimodal profile suggests superiority over narrow glucose-lowering agents, aligning it with modern therapeutic strategies that target both metabolic and inflammatory pathways. However, the present study is limited by its short duration, relatively small sample size, and reliance on an alloxan model, which mimics type 1-like β-cell injury rather than type 2 diabetes. Future investigations in high-fat diet or genetic models, alongside pharmacokinetic and chronic toxicity studies, are warranted to establish translational potential.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

ETHICAL APPROVAL:

All experimental procedures were approved by the Institutional Animal Ethics Committee (IAEC/REETESH KUMAR RAIVU/PhD/ PRMOCI7B02/KMCP/106/2020-2) and conducted in accordance with CPCSEA guidelines.

ABBREVIATIONS:

AIP – Atherogenic Index of Plasma, AI – Atherogenic Index, ALT – Alanine aminotransferase, AMPK – AMP-activated protein kinase, ANOVA – Analysis of Variance, AST – Aspartate aminotransferase, CAT – Catalase, COI – Conflict of Interest, CRP – C-reactive protein, CRI – Coronary Risk Index, CPCSEA – Committee for the Purpose of Control and Supervision of Experiments on Animals, DNA – Deoxyribonucleic acid, DTNB – 5,5’-Dithiobis-(2-nitrobenzoic acid), ELISA – Enzyme-Linked Immunosorbent Assay, FBG – Fasting Blood Glucose, GSH – Reduced Glutathione, HbA1c – Glycated Hemoglobin A1c, HDL – High-Density Lipoprotein, HandE – Hematoxylin and Eosin, HOMA-IR – Homeostasis Model Assessment for Insulin Resistance, HOMA-β – Homeostasis Model Assessment for β-cell function, IAEC – Institutional Animal Ethics Committee, ILI – Inflammatory Load Index, IL-6 – Interleukin-6, LDL – Low-Density Lipoprotein, LPS – Lipopolysaccharide, MDA – Malondialdehyde, NF-κB – Nuclear Factor kappa-light-chain-enhancer of activated B cells, p.o. – Per os (oral administration), ROS – Reactive Oxygen Species, SEM – Standard Error of Mean, SOD – Superoxide Dismutase, SPSS – Statistical Package for the Social Sciences, STZ – Streptozotocin, TC – Total Cholesterol, TG – Triglycerides, TNF-α – Tumor Necrosis Factor-alpha, VLDL – Very Low-Density Lipoprotein.

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Received on 12.12.2024 Revised on 05.07.2025

Accepted on 07.11.2025 Published on 10.02.2026

Available online from February 16, 2026

Research J. Pharmacy and Technology. 2026;19(2):811-819.

DOI: 10.52711/0974-360X.2026.00116

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

Group	Liver Glycogen (mg/g)	Hexokinase (U/mg protein)	Glucokinase (U/mg protein)	Glucose-6-phosphatase (U/mg protein)	% Change vs Diabetic (Glycogen)
Normal Control	48.35 ± 3.50	0.218 ± 0.014	30.58 ± 1.42	0.390 ± 0.018	↑216%
Diabetic Control	15.30 ± 0.72^a	0.105 ± 0.009^a	7.50 ± 0.36	0.125 ± 0.008^a	Reference
Glipizide (10 mg/kg)	40.65 ± 2.95^b	0.165 ± 0.013^b	22.30 ± 0.90^b	0.315 ± 0.016^b	↑166%
Pyridazinone 30 mg/kg	33.60 ± 1.85^b	0.140 ± 0.008^b	17.45 ± 0.63^b	0.260 ± 0.010^b	↑120%
Pyridazinone 60 mg/kg	36.75 ± 2.05^b	0.156 ± 0.012^b	19.80 ± 0.72^b	0.276 ± 0.012^b	↑140%