INTRODUCTION:

Diabetes mellitus is a significant metabolic condition. Chronic hyperglycaemia may result from¹ insufficient insulin synthesis or ² the inability of peripheral tissues to respond to insulin. Diabetes mellitus proliferates swiftly. In 2019, 9.3% of the worldwide adult population was diagnosed with diabetes. In 2017, the International Diabetes Federation reported that 352 million individuals were at risk of type 2 diabetes.

Health predictions indicate that by 2030, 439 million persons would be impacted by diabetes^3,4. Individuals afflicted with diabetes mellitus in hyperglycaemic conditions generate free radicals. Furthermore, these free radicals are neutralized by antioxidants⁵. In diabetes mellitus, hyperglycaemic patients have a reduction in their enzymatic antioxidants. The enzyme superoxide dismutase (SOD) is crucial for safeguarding β-cells against the detrimental effects of reactive oxygen species (ROS) and free radical assault. The chemicals assist in avoiding several consequences of diabetes mellitus and neutralizing free radicals^6,7. The suppression of reactive oxygen species (ROS) by the flavonoid’s quercetin and rutin represents optimal bioactive substances that assist in neutralizing free radicals and are particularly effective against cardiovascular and diabetes problems⁸. Flavonoids possess significant antioxidant properties, safeguarding the body from damage inflicted by reactive oxygen species by donating hydrogen atoms and binding with free radicals to create stable flavonoid radicals. Consequently, it can impede lipid peroxidation damage to cells and avert consequences such as diabetic mellitus ^9.

Aziridines are three-membered ring heterocycles containing nitrogen, recognized for their role as reactive intermediates in the synthesis of chiral amino alcohols, azomethine ylides, and amino acid derivatives ¹⁰. Additionally, they serve as chiral ligands and auxiliaries in asymmetric synthesis of fused heterocycles ^11,12. Aziridine-containing compounds serve as significant reactive intermediates and exhibit many biological activities, notably antibacterial and anticancer properties, due to the presence of the aziridine ring ¹³.

Fig 1. Chemical structure of Aziridine derivatives

Aziridines are potent alkylating agents, and their in-vivo efficacy is predominantly attributed to toxicity rather than particular action. The physiological effect of mitomycin C depends on the opening of the aziridine ring and its interaction with the guanine nucleobase of DNA during the alkylation reaction ^14. This results in the production of covalent interstrand DNA–DNA crosslinks, inhibition of replication, and ultimately cell death. Aziridines containing the amide functional group are presently of particular interest, with Imexon being a notable example ^15. Imexon is an anticancer treatment particularly effective against human myeloma cells, where it binds to cellular thiols, diminishes the levels of glutathione and cysteine in target cells, resulting in increased concentrations of reactive oxygen species (ROS).

MATERIALS AND METHODS:

Materials:

Aziridine derivatives, ethanol, albino mice, alloxan, Wistar rats, glibenclamide digital weighing balance, distilled water.

Preparation of animals:

Albino rats (either sex) weighing 130–150g were provided by Animal House, Institute of Pharmaceutical Sciences and Research, Unnao. They were housed in healthy conditions, with a 12-hour light/dark cycle and a room temperature of 25°C. The rats were given a typical rodent food and unrestricted access to water, and the relative humidity is maintained between 44 and 56 percent. Up to one hour before to treatment exposures, the animals were kept on fasting while still having unrestricted access to water ^16.

Acute toxicity (LD₅₀) study:

The acute toxicity (dose regimen) test of the synthesized aziridine derivatives was determined using Lorke’s method (1983). The synthesized aziridine derivatives are prepared in DMSO (10%v/v). The test was carried out in two phases. The first phase involved the use of 9 Wistar rats randomly distributed into 3 groups. The groups are administered 10, 100 and 1000mg/kg body weight of the synthesized aziridine derivatives orally. The animals are monitored for 24 hours for gross behaviour and mortality. The second phase involved 3 Wistar rats. Three different doses of 1600, 2900 and 5000mg/kg body weight of the synthesized aziridine derivatives are administered to each rat. The animals are monitored for 24 hours for mortality. The LD₅₀ was calculated as the geometric mean of the maximum dose that caused 0% death and the minimum dose that caused 100% death ¹⁷.

Induction of experimental diabetes:

Male Swiss albino mice were fasted overnight (12–14h) and their weight and fasting blood glucose level was recorded with a glucometer and then made diabetic by a single intraperitoneal injection (a volume of 1ml/kg) of freshly prepared alloxan monohydrate solution (20mg/kg body weight). Alloxan was prepared by weighing according to individual animal weight and solubilized with 0.5ml sodium citrate at pH 4.5 before injection. Food and water are presented to the animals 30 min after administration of alloxan. After 48 h of alloxan injection, plasma blood glucose level of each animal was determined by taking the blood from the tail and animals with a fasting blood glucose level above 200mg/dl are included in the study ^18.

Group design:

All the mice were divided into 5 groups (n=6) and treated for 21 days[20] as follows-

Group 1 given normal saline [control].

Group 2 given alloxan (20mg/kg, p. o.) [disease control].

Group 3 given alloxan (20mg/kg, p. o.) + glibenclamide (10mg/kg) [Std.].

Group 4 given alloxan (20mg/kg, p. o.) + synthesized aziridine derivatives (50mg/kg, p. o.) [Test 1].

Group 5 given alloxan (20mg/kg, p. o.) + synthesized aziridine derivatives (100mg/kg, p. o.) [Test 2].

Pharmacological evaluation

· Evaluation of antioxidant potential

1. Estimation of DPPH Radical Scavenging Assay:

The free radical scavenging capacity of synthesized aziridine derivatives was assessed in vitro using the 1,1′-diphenyl-1-picrylhydrazyl (DPPH) technique as described by Tariq et al.¹⁹. The assay involved the reaction of 1.6 mL of 0.135 mM DPPH dissolved in 100% v/v methanol with 0.4 mL of various concentrations (0.0078–2 mg/mL) of produced aziridine derivatives. The reaction mixture was vigorously vortexed and then incubated in the dark at ambient temperature for 30 minutes. The absorbance of the combination was measured at 517 nm after 2 minutes. The concentration of the synthesized aziridine derivatives required to reduce the absorbance of DPPH radical by 50% was determined. Rutin (Sigma-Aldrich, ≥94%, HPLC grade) is utilized as a reference medication at identical working concentrations to the produced aziridine derivatives.

The % scavenging activity was determined by the following equation:

DPPH scavenging activity (%)= [(control absorbance − sample absorbance)/(control absorbance)] × 100

2. Estimation of total antioxidant activity:

The approach employed by Prieto et al^{. 20} was utilized to assess the total antioxidant activity of the compounds. A little quantity of derivative, 4 mM ammonium molybdate, 28 mM sodium phosphate, and 0.3 ml sulfuric acid are combined. The reaction mixture was incubated at 95°F for 90 minutes in a water bath. The absorbance of each sample combination is measured at 695 nm. Total antioxidant activity is determined by measuring the ascorbic acid equivalents in milligrams per gram of derivatives.

· Evaluation of anti-diabetic potential:

1. Estimation of α-amylase:

The alpha-amylase assay was conducted following the procedure outlined by Odeyemi ^21. Fifteen microliters of synthetic aziridine derivatives at varying concentrations (50μg/ml – 200μg/ml), diluted in phosphate buffer, were added to five microliters of porcine pancreatic enzyme solution on a 96-well plate. Following a 10-minute incubation at 37°C, the reaction commenced with the addition of 20μl of starch solution, which was subsequently incubated for an additional 30 minutes at 37°C. The reaction was subsequently halted by the addition of 10μl of 1M HCl to each well, followed by 75μl of iodine reagent. A blank with phosphate buffer (pH 6.9) in lieu of the synthesized aziridine derivatives and a positive control (acarbose, 64μg/ml) were created. No enzyme control and no starch control were incorporated for each test sample²². The absorbance was recorded at 580 nm, and the percentage of inhibitory activity was determined using the subsequent equation.:

Inhibition (%) = 100-% reaction (at min), where % reaction = mean maltose in sample × 100/mean maltose in control.

2. Estimation of α-glucosidase:

To 50 µL of each derivative concentration in a test tube (0-40 µg/mL), the following were added sequentially: Buffered α-glucosidase (100 µL, 1.0 U/mL) was incubated at 37°C for 10 minutes, followed by the addition of pNPG (50 µL, 3.0 mM) and further incubation at 37°C for 20 minutes²³. Subsequently, Na2CO3 (5% w/v) was added, the mixture was cooled to 25°C, and finally, 5 mL of H2O was incorporated and vortexed ²⁴. The absorbance of yellow p-nitrophenol from several test tubes was measured at 405 nm, and the % inhibition was computed accordingly:

% Inhibition= Ac−At/Ac × 100

3. Body weight:

All animals in each group are weighed prior to drug delivery and after the dose is completed. The body weight is compared prior to and following drug delivery.

4. Blood sugar level:

Blood glucose levels were measured seven times at 0, 5, 10, and 15 days following the initiation of medication treatment. A blood sample is obtained from the punctured tail vein, and blood glucose levels are measured using a glucometer manufactured by Dr. Morepen. This process is simple and genuine ^25.

5. Oral glucose tolerance test (OGTT):

Following two weeks of treatment with the produced aziridine derivatives, the animals were undergone a fasting period of 12–14 hours while having unrestricted access to water, during which their fasting blood glucose levels were tested four times. A glucose solution (2g/kg of body weight) was supplied orally at a volume of 1 mL/kg. Blood samples were obtained at 30, 60, and 120 minutes post-glucose delivery ^26.

RESULT:

A. Determination of acute toxicity (LD₅₀) of aziridine derivative:

In phase I and phase II, all the animals were found safe with no behavioural signs at all the dose levels. No mortality was seen in any animal. Thus, aziridine derivatives are safe to use in the dose range of 100-5000mg/kg. Thus, oral acute toxicity (LD₅₀) is >5000mg/kg.

Table 1. Determination of acute toxicity (LD₅₀)

Phase	No. of rats	Dose (mg/kg)	Behavioural signs	Mortality
I	03	10	None	0/3
I	03	100	None	0/3
I	03	1000	None	0/3
II	01	1600	None	0/1
II	01	2900	None	0/1
II	01	5000	None	0/1

B. Evaluation of antioxidant potential:

1. DPPH Scavenging capacity:

At 800µg/ml conc., aziridine derivatives (A1 and A4) showed the % inhibition in DPPH scavenging assay capacity as 93.42±0.52%, 94.26±0.18%, respectively which was statistically significant when compared with control. However, ascorbic acid showed highest percent inhibition as 68.23±0.40% and 97.22±0.49%, at the conc. of 400µg/ml and 800µg/ml, respectively. The antioxidant action might be due to suppression of free radical formation.

The % inhibition of DPPH Scavenging assay was observed as follow-

Table 2. DPPH Scavenging capacity

Substance	DPPH Scavenging capacity (% Inhibition) 200µg/ml 400µg/ml 800µg/ml
Water	17.24±0.12	19.10±0.45	18.45±0.21
Ascorbic acid	38.34±0.23	68.23±0.40	97.22±0.49
A1	33.45±0.39	65.27±0.23	93.42±0.52
A4	35.29±0.37	65.87±0.65	94.26±0.18

Fig 2. Determination of DPPH Scavenging capacity

2. Total antioxidant activity:

Both the aziridine derivatives showed total antioxidant activity in contrast to water. A1 and A4 exhibited the total antioxidant activity as 63.49±0.18% and 65.78±0.34%, respectively at the conc. of 400µg/ml. However, at 800µg/ml, A1 and A4 demonstrated the increasing total antioxidant activity as 76.67±0.23% and 78.39±0.19%, respectively. Negligible antioxidant response was observed in the water/control. Therefore, aziridine derivatives might show antioxidant response by minimizing the oxidation or production of ROS.

Table 3. Total antioxidant activity

Substance	Total antioxidant activity 200µg/ml 400µg/ml 800µg/ml
Water	22.46±0.25	25.18±0.45	24.56±0.22
Ascorbic acid	42.49±0.32	72.53±0.11	92.31±0.39
A1	31.45±0.17	63.49±0.18	76.67±0.23
A4	33.20±0.12	65.78±0.34	78.39±0.19

Fig 3. Graphical data of total antioxidant activity

C. Evaluation of anti-diabetic activity:

1. Estimation of α-amylase inhibition:

At higher conc. (800µg/ml), the α-amylase inhibition was estimated as 82.39±0.16%, and 83.58±0.13%, in A1 and A4, respectively. The inhibition rate was observed in dose-dependent manner. Therefore, it can be easily observed that aziridine derivatives significantly lowered the hyperglycaemia level as it showed an effective α-Amylase inhibition when compared with control.

Table 4. Estimation of α-amylase inhibition of novel derivatives of aziridine

Substance	α-Amylase inhibition (% Inhibition) 200µg/ml 400µg/ml 800µg/ml
Water	24.36±0.12	26.42±0.37	25.22±0.16
Acarbose	49.54±0.10	69.25±0.28	91.45±0.20
A1	37.17±0.28	57.26±0.16	82.39±0.16
A4	38.46±.30	58.29±0.25	83.58±0.13

Fig 4. Estimation of α-amylase inhibition of novel derivatives of aziridine

2. Estimation of α-glucosidase inhibition:

A1 and A2 showed the prominent α-glucosidase inhibition at all the concentrations utilized. At 800µg/ml, the % inhibition was observed as 74.33±0.25%, and 78.16±0.32%, in aziridine derivatives (A1 and A4), respectively. Acarbose showed highest % inhibition at lower and higher concentrations.

Table 5. Estimation of α-glucosidase inhibition of novel derivatives of aziridine

Substance	α-glucosidase inhibition (% Inhibition) 200µg/ml 400µg/ml 800µg/ml
Water	24.36±0.17	27.53±0.24	26.39±0.14
Acarbose	47.27±0.25	74.20±0.14	93.56±0.27
A1	33.41±0.13	45.36±0.18	74.33±0.25
A4	35.34±.22	47.25±0.19	78.16±0.32

Fig. 5. Graphical data of α-glucosidase inhibition of novel derivatives of aziridine

3. Body weight measurements:

Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.) and alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.) treated mice showed body weight lowering effect when compared with the body weight of mice (before treatment). Alloxan (20mg/kg) treated mice showed the highest weight gain. Thus, both the aziridine derivatives (A1 and A4) showed the anti-diabetic potential when compared with the normal saline treated animals.

Table 6. Body weight measurements of aziridine derivatives treated mice

Treatment	Body weight (g) Before After
Normal saline	38.13±0.18	39.45±0.24
Alloxan (20mg/kg, s. c.)	36.17±0.24	53.17±0.24
Alloxan (20mg/kg, s. c.) + Glibenclamide (5mg/kg, i. p.)	40.25±0.12	44.25±0.12
Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.)	38.53±0.27	49.53±0.27
Alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.)	39.27±0.38	47.27±0.38

Significant data at P≤0.05 and n=6

Data demonstrated as Mean±SEM

Fig 6. Graphical data of body weight measurements of aziridine derivatives treated rats

4. Estimation of blood sugar level:

The blood sugar level was estimated after 0, 5, 10, 15 and 21 days. After 21 days, Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.) and alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.) treated rats showed the blood glucose level as 129.15±0.26mg/dl and 126.24±0.16mg/dl, respectively. However, maximum glucose level was observed in disease control group as 186.21±0.36mg/dl. Therefore, at both the aziridine derivatives showed the anti-hyperglycemic action. A decrease in blood glucose levels is indicative of the hypoglycemic properties of the aziridine derivatives.

Fig 7. Graphical data of estimation of blood sugar level of aziridine treated mice

5. Oral glucose tolerance test:

In oral glucose tolerance test, the blood sugar level was found increasing over time. However, after 120 min it was lowered due to better adaptability and glucose utilization. After 120 min, Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.) and Alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.) treated mice showed blood glucose level as 137.12±0.38mg/dl and 134.36±0.29mg/dl, respectively.

Table 8. Estimation of blood sugar level in oral glucose tolerance test

Treatment	Blood sugar level (mg/dl) in OGTT 30 min 60 min 120 min
Normal saline	80.26±0.29	86.37±0.49	92.23±0.56
Alloxan (20mg/kg, s. c)	88.53±0.15	93.39±0.27	86.57±0.35
Alloxan (20mg/kg, s. c.) + Glibenclamide (5mg/kg, i. p.)	104.27±0.56	114.35±0.29	123±0.63
Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.)	116.41±0.33	128.17±0.28	137.12±0.38
Alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.)	112.16±0.20	126.24±0.15	134.36±0.29

Significant data at P≤0.05 and n=6

Data demonstrated as Mean±SEM

Fig 8. Graphical data of blood sugar level in oral glucose tolerance test

A total of 12 new aziridine derivatives were synthesized and evaluated regarding their physicochemical features and spectrum analysis. In the docking investigation, molecules A1 and A4 exhibited the greatest binding affinity among the 12 aziridine derivatives. Thus, A1 and A4 were utilized for the evaluation of antioxidant and anti-diabetic potential using both in-vitro and in-vivo models. This suggests that insulin control is normalized or that sensitivity to tyrosine kinase receptor subtypes is increased, hence enhancing binding and activating glucose transporters. Thus, it promotes the liberation of glucose molecules for improved delivery to target organs and produces ATP energy for optimal metabolic processes in tissues. The mechanism of action may depend on insulin secretion in Type 1 diabetes mellitus or receptor sensitization in Type 2 diabetes mellitus.

CONCLUSION:

It concluded that aziridine derivatives (A1 and A4) were demonstrated as significant antioxidant potential in DPPH scavenging assay and total antioxidant activity. Aziridine derivatives were found safe in LD₅₀study i.e., till 5000mg/kg dose, 0/3 mortality was observed. Both the derivatives showed the prominent percentage inhibition in α-amylase and α-glucosidase. When observed in animal study, A1 and A4 exhibited the hypoglycaemic action by lowering the blood glucose level and body weight of the mice. It suggests that aziridine derivatives (A1 and A4) are promising moieties which might be utilized in the management of oxidation and associated diabetes mellitus.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

REFERENCES:

1. Amit Sapra, Priyanka Bhandari. Diabetes. Treasure Island (FL): StatPearls Publishing; 2024.

2. Rajaei E, Jalali MT, Shahrabi S, Asnafi AA, Pezeshki SMS. HLAs in Autoimmune Diseases: Dependable Diagnostic Biomarkers? Curr Rheumatol Revi. 2019; 15(4): 269-276. [PubMed]

3. Alem Geberemeskel G., Godefa Debebe Y., Abraha Nguse N Clinical study antidiabetic effect of fenugreek seed powder solution (Trigonella foenum-graecum L.) on hyperlipidemia in diabetic patients. J. Diabetes Res. 2019; 8507453

4. Shaw J.E., Sicree R.A., Zimmet P.Z, Global estimates of the prevalence of diabetes. Diabetes Res. Clin. Pract 2010; 87; 4-14.

5. Ratnaningtyas NI, Hernanyanti, Ekowati N, et al. Antioxidant activities and properties of Coprinuscomatus mushroom both mycelium and fruiting body extracts in streptozotocin-induced hyperglycemic rats model. Biosaintifika Journal of Biology and Biology Education. 2022; 14(1); 9- 21.

6. Bhat IUH, Bhat R. Quercetin: A bioactive compound imparting cardiovascular and neuroprotective benefits: Scope for exploring fresh produce, their wastes, and byproducts. Biology (Basel). 2021; 10(7): 1-34.

7. Ratnaningtyas NI, Hernayanti H, Ekowati N, Husen F. Ethanol extract of the mushroom Coprinuscomatus exhibits antidiabetic and antioxidant activities in streptozotocin-induced diabetic rats Nuniek. Pharm Biol. 2021; 60(1); 1126-1136.

8. Kaneto H, Kimura T, Shimoda M, et al. Favorable effects of GLP-1 receptor agonist against pancreatic β-cell glucose toxicity and the development of arteriosclerosis: “the earlier, the better” in therapy with incretin-based medicine. Int J Mol Sci. 2021; 22(15); 7917.

9. Husen F, Hernayanti H, Ekowati N, Sukmawati D, Ratnaningtyas NI. Antidiabetic effects and antioxidant properties of the saggy ink cap medicinal mushroom, Coprinuscomatus (Agaricomycetes) on streptozotocin-induced hyperglycemic rats. Int J Med Mushrooms. 2021; 23(10); 9-21.

10. Yudin, A.K. Aziridines and Epoxides in Organic Synthesis; Wiley-VCH: Weinheim, Germany. 2006; 494.

11. Morieux, P.; Stables, J.P.; Kohn, H. Synthesis and anticonvulsant activities of N-benzyl (2R)-2-acetamido-3-oxysubstituted propionamide derivatives. Bioorg. Med. Chem. 2006; 16; 8968–8975.

12. Cimarelli, C.; Fratoni, D.; Palmieri, G. A Convenient Synthesis of New Diamine, Amino Alcohol and Aminophosphines Chiral Auxiliaries Based on Limonene Oxide. Tetrahedron Asymmetry 2009; 20; 2234–2239.

13. Le´sniak, S.; Pieczonka, A.M.; Jarzy ´nski, S.; Justyna, K.; Rachwalski, M. ChemInform Abstract: Synthesis and Evaluation of the Catalytic Properties of Semicarbazides Derived from N-Triphenylmethyl-aziridine-2-carbohydrazides. Tetrahedron Asymmetry. 2013; 24; 1341–1344.

14. Pieczonka, A.M.; Le´sniak, S.; Rachwalski, M. Direct asymmetric aldol condensation catalyzed by aziridine semicarbazide zinc(II) complexes. Tetrahedron Lett. 2014; 55; 2373–2375.

15. Dvorakova, K.; Waltmire, C.N.; Payne, C.M.; Tome, M.E.; Briehl, M.M.; Dorr, R.T. Induction of mitochondrial changes in myeloma cells by imexon. Blood. 2001; 97; 3544–3551.

16. Sheveleva, E.V.; Landowski, T.H.; Samulitis, B.K.; Bartholomeusz, G.; Powis, G.; Dorr, R.T. Imexon Induces an Oxidative Endoplasmic Reticulum Stress Response in Pancreatic Cancer Cells. Mol. Cancer Res. 2012; 10; 392–400.

17. Bhajoni et al. Evaluation of the Antiulcer Activity of the Leaves of Azadirachta indica: An Experimental Study. Integrative Medicine International. 2016; 3: 10–16.

18. Nyarko AK, Asare-Anane H, Ofosuhene M, Addy ME. Extract of Ocimum canum lowers blood glucose and facilitates insulin release by isolated pancreatic b-islet cells. Phytomed 2002; 9; 346–51.

19. Carvalho EN, Carvalho NAS, Ferreira LM. Experimental model of induction of diabetes mellitus in rats. Acta Cir Bras. 2003; 18; 60–64.

20. Nagappa AN, Thakurdesai PA, Rao NV, Singh J. Antidiabetic activity of Terminalia catappa Linn fruits. J Ethnopharmacol. 2003; 88(1); 45–50.

21. Tariq A, M. Athar, J. Ara, V. Sultana, S. Ehteshamul-Haque, and M. Ahmad. Biochemical evaluation of antioxidant activity in extracts and polysaccharide fractions of seaweeds, Global Journal of Environmental Science Management, vol. 2015; 1; 47–62.

22. Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem. 1999; 269: 337-41.

23. Odeyemi S. W. A comparative study of the in vitro antidiabetic properties, cytotoxicity and mechanism of action of Albuca bracteata and Albuca setosa bulb extracts Doctoral dissertation, University of Fort Hare, 2015.

24. Chimaobi J. Ononamadu, Adamu J. Alhassan, Abdullahi A. Imam, Aminu Ibrahim, Godwin O. Ihegboro, Alowonle T. Owolarafe, and Mohammed S. Sule. (2019) In vitro and in vivo anti-diabetic and anti-oxidant activities of methanolic leaf extracts of Ocimum canum. Caspian J Intern Med. 2008; 10(2); 162–175.

25. Jung Jae Hyun, Su Bin Hwang, Hyeon Ju Park, Guang-Ri Jin, and Bog Hieu Lee. Anti-obesity and Antidiabetic Effects of Portulaca oleracea Powder Intake in High-Fat Diet-Induced Obese C57BL/6 Mice. Evidence-Based Complementary and Alternative Medicine. 2021, Article ID 5587848: 1-11.

26. Nagappa AN, Thakurdesai PA, Rao NV, Singh J Antidiabetic activity of Terminalia catappa Linn fruits. J Ethnopharmacol. 2003; 88(1); 45–50.

Received on 03.06.2024 Revised on 13.08.2024

Accepted on 23.10.2024 Published on 24.12.2024

Available online from December 27, 2024

Research J. Pharmacy and Technology. 2024;17(12):6118-6124.

DOI: 10.52711/0974-360X.2024.00928

Treatment	Blood sugar level (mg/dl) 0 days 5 days 10 days 15 days 21 days
Normal saline	82.34±0.23	84.17±0.18	81.67±0.54	86.34±0.20	82.50±0.14
Alloxan (20mg/kg, s. c)	84.34±0.31	102.49±0.16	138.36±0.42	182.25±0.14	186.21±0.36
Alloxan (20mg/kg, s. c.) + Glibenclamide (5mg/kg, i. p.)	86.21±0.34	89.67±0.13	92.14±0.28	96.28±0.20	94.24±0.15
Alloxan (20mg/kg, p. o.) + aziridine derivative- A1 (50mg/kg, p. o.)	82.45±0.16	86.16±0.27	96.23±0.18	108.45±0.27	129.15±0.26
Alloxan (20mg/kg, p. o.) + aziridine derivative- A4 (50mg/kg, p. o.)	85.37±0.53	91.56±0.55	103.36±0.33	117.31±0.12	126.24±0.16