Synthetic Novel Flavanoid derivatives act as potential Antidiabetic agent against Streptozocin induced in diabetic Rats

 

G. Babu¹, Asish Bhaumik^2*, K. Krishnamachary³, P. Kalyani⁴

¹Assistant Professor, Department of Pharmaceutical Chemistry, Anurag Pharmacy College,

Ananthagiri, Kodad, Suryapet-508206, Telangana State, India

²Associate Professor, Department of Pharmaceutical Chemistry, Anurag Pharmacy College,

Ananthagiri, Kodad, Suryapet-508206, Telangana State, India

³Department of Pharmaceutical Analysis and Quality Assurance, Sree Dattha Institute of Pharmacy,

Sheriguda, Ibrahimpatnam, Hyderabad-501510, Telangana State, India.

⁴Assistant Professor, Department of Pharmaceutical Analysis, Anurag Pharmacy College,

Ananthagiri, Kodad, Suryapet-508206, Telangana State, India

 

ABSTRACT:

Diabetes mellitus is a heterogeneous group of diseases characterized by chronic elevation of glucose in the blood. It arises because the body is unable to produce enough insulin for its own needs, either because of impaired insulin secretion, impaired insulin action, or both. Chronic exposure to high blood glucose is a leading cause of renal failure, visual loss and a range of other types of tissue damage. Diabetes also predisposes to arterial disease, not least because it is often accompanied by hypertension, lipid disorders and obesity. Many cases of diabetes and almost all of its unwanted long-term consequences are potentially avoidable, but this will require intervention at a societal as well as at a medical level. The three classic symptoms of diabetes are thirst, polyuria and weight loss. As glucose is lost in the urine it draws fluid and other small molecules with it, causing excessive urination, which in turn causes dehydration and thirst. Weight is lost because of rapid breakdown of fat and protein reserves to compensate for the loss of glucose and metabolic inefficiency due to lack of insulin action. These symptoms may be less prominent in older people with type 2-diabetes, who may present with symptoms less directly related to diabetes, or with complications of diabetes ranging from infections to heart disease, or simply as the result of a screening blood test. The objective of the present research work was the synthesis of 2- (2, 3, 4, & 5 substituted phenyl) 3-hydroxy-4H-Chromen-4-one and evaluation of in vivo anti diabetic activity against streptozocin induced diabetic Rats. Based on this a new series of compound had been planned to synthesize by reacting 2-hydroxy acetophenone and various aromatic aldehydes in the  presence of potassium hydroxide, methanol and 30% hydrogen peroxide. The synthesized compounds were characterized by IR, NMR, and Mass spectroscopy. The present experimental data of blood sugar, serum lipid profile and urinary data displayed that all the synthesized compounds had the potential ability to reduce the diabetes mellitus which was induced by streptozocin in rats.

 

 

 

 

INTRODUCTION:

Flavonoids (or bioflavonoids) (from the Latin word flavus meaning yellow, their color in nature) are a class of plant and fungus secondary metabolites. Chemically, they have the general structure of a 15-carbon skeleton, which consists of two phenyl rings (A and B) and heterocyclic ring (C). This carbon structure can be abbreviated C6-C3-C6. According to the IUPAC nomenclature [1, 2] they can be classified into: flavonoids or bioflavonoids, isoflavonoids, derived from 3-phenylchromen-4-one (3-phenyl-1,4-benzopyrone), neoflavonoids, derived from 4-phenylcoumarine (4-phenyl-1,2-benzopyrone) structure.

 

Table-1: Molecular structure of the flavone backbone

2-phenyl – 4H-chromen-4-one	3-phenyl-3,4-dihydro-2H-1-benzopyran		4-phenyl-2H-chromen-2-one
2-phenyl – 4H-chromen-4-one	2-phenyl-3,4-dihydro-2H-1-benzopyran-3,4-diol		3-hydroxy-2-phenyl-3,4-dihydro-2H-1-benzopyran-4-one
2-phenyl-3,4-dihydro-2H-1-benzopyran		2-phenyl-3,4-dihydro-2H-1-benzopyran-3-ol
3-hydroxy-2-phenyl-3,4-dihydro-2H-1-benzopyran-4-one		2-phenyl-3,4-dihydro-2H-1-benzopyran-4-ol

 

The three flavonoid classes above are all ketone-containing compounds, and as such, are anthoxanthins (flavones and flavonols). This class was the first to be termed bioflavonoids. The terms flavonoid and bioflavonoid have also been more loosely used to describe non-ketone polyhydroxy polyphenol compounds which are more specifically termed flavanoids. The three cycle or heterocycles in the flavonoid backbone are generally called ring A, B and C. Ring A usually shows a phloroglucinol substitution pattern.

 

Flavonoids possessed a wide range of biological and pharmacological activities in in vitro studies. Examples include anti-allergic[3], anti-inflammatory[3, 4], antioxidant[4], anti-microbial(antibacterial[5, 6] antifungal [7,8] and antiviral[7,8]), anti-cancer [9] and anti-diarrheal activities [10]. Flavonoids have also been shown to inhibit topoisomerase enzymes [11, 12] and to induce DNA mutations in the mixed-lineage leukemia (MLL) gene in in vitro studies [13]. However, in most of the above cases no follow up in vivo or clinical research has been performed, leaving it impossible to say if these activities have any beneficial or detrimental effect on human health.

 

EXPERIMENTAL CHEMISTRY:

Materials and method:

Drugs and chemicals used:

Glibenclamide, streptozotocin (STZ), and sodium citrate buffer were used in this study. Other chemicals used for the synthesis of targets compounds were provided by institutional store and all were of AR and LR grade.

 

Instruments:

The progress of the reaction was monitored by TLC using solvent systems of different polarities. TLC plates are pre-coated silica gel (HF254-200 mesh) aluminium and spots were visualized under U.V chamber. The melting point of newly synthesized flavanoid compounds were determined by open capillary method. The IR spectra of synthesized compounds were recorded by ABB Bomen FT-IR spectrometer MB 104 IR spectra recorder with KBr pellets. The H¹-NMR spectra of synthesized compounds were obtained from Bruker Avance II 400 MHz spectrometer using TMS as an internal standard in CDCl₃. All the Mass spectra (MS, HR-MS) of synthesized compounds were recorded on a LTQ-orbitrap linear ion trap high resolution mass spectrometer. The IR, H¹-NMR and Mass spectra were assigned to elucidate the structure of synthesized compounds (4A-4J).

 

Synthetic scheme [14]:

 

General procedure for the synthesis of target compounds:

2-hydroxy acetophenone [(1), 1.36gm, 0.01mol] and benzaldehyde [(2), 1.06gm, 0.01mol) were added to a solution of potassium hydroxide (1.12gm, 0.02 mol) in methanol (50 ml) at 0-5^oC. The reaction mixture was stirred over night at room temperature and then poured over crushed ice and acidified to P^H 6 with 2M Hcl. The resulting yellow solid was filtered and the filter cake washed with water to give the crude product that either be crystallized from ethanol to afford pure 2-hydroxy chalcones 3A or used directly in the next reaction without further purification.

 

30% hydrogen peroxide (10 ml) was added to a well stirred solution of 3A (1.57gm, 0.007 mol) and 20% (w/w) aqueous potassium hydroxide (10 ml) in methanol (20 ml) at 0-5^oC in a drop wise manner over 1 hour. The resulting reaction mixture was stirred for 10 hours and then poured on crushed ice and neutralized with 2M Hcl. Ethyl aceto acetate (50 ml) was added and the organic layer was washed successfully with water, a saturated solution of sodium bicarbonate, water and brine and then dried over anhydrous magnesium sulphate. The solvent was removed in vacuum and the residue was purified by column chromatography on silica gel (ACOEt/ n-hexane = 1/3 to 1/1) to give the title compound 4A as a white solid.

 

EXPERIMENTAL PHARMACOLOGY:

Experimental animals:

White male albino Wister rats weighing about 200-250gm was used, they were obtained from the animal house of C. L. Baid Metha College of Pharmacy, chennai. They were kept under observation for about 7 days before onset of experiment to exclude any intercurrent infection, had free access to normal diet and water. The experimental protocol was approved by IAEC (Institutional Animal Ethics Committee) of CPCSEA: IAEC/XXIX/12/2015.

 

Evaluation of acute oral toxicity [15]:

In the present study the acute oral toxicity of the synthesized compounds were performed by acute toxic class method. In this method the toxicity of the compound was planned to test using step wise procedure, each step using three Wister rats. The rats were fasted prior to dosing (food but not water should be withheld) for three to four hrs. Following the period of fasting the animals were weighed and the compound was administered orally at a dose of 2000 mg/Kg b. w. Animals were observed individually after dosing at least once during the first 30 min; periodically the surveillance was carried out for the first 24 hrs with special attention given during the first 4 hrs and daily thereafter, for a total of 14 days. No animal died. Therefore, the LD₅₀ is greater than 2000 mg/kg. An investigation with 1/20th, 1/10th, and 1/5th of 2000 mg/ kg, i.e. 100, 200, and 400 mg was done in pre-screening. Only 400 mg/kg was found to be effective against diabetes, hence this dose was used in final screening. The experimental protocol was approved by Institutional Animal Ethics Committee (IAEC).

 

Evaluation of Anti diabetic Activity [16, 17, 18]:

After fasting, DM was induced by intra peritoneal injection of STZ dissolved in 0.1 M cold sodium citrate buffer (pH 4.4) at a dose of 30 mg/kg b. w. The animals were allowed to drink 5% glucose solution overnight to overcome the drug-induced hypoglycemia. After 72 h, STZ-treated animals were considered as diabetic when the fasting plasma levels were observed above 200 mg/dL with glycosuria. The experiment was carried out with four groups of animals and five rats were used in each of the four groups which were given as below:

A   Group I: Normal control (vehicle).

B   Group II: Diabetic control (vehicle).

C   Group III: Diabetic rats treated with synthesized compounds (4A-4J) (400 mg/kg p. o.)

D   Group IV: Diabetic rats treated with glibenclamide (5 mg/kg p. o.)

 

Vehicle, synthesized compounds, and glibenclamide were administered once daily for 21 days from the day of induction. Blood was drawn from tip of the tail, and blood glucose level was estimated on 0, 7th, 14th, and 21st day of experiment with the help of glucometer using strip method. On 21st day, blood sample was collected by retro orbital puncture for measuring serum cholesterol and TG level using auto analyser. Fresh urine was collected for the determination glucose and ketone bodies, by using keto-diastix strips on 0 and 21st day of the experiment.

 

Statistical analysis:

All results were expressed as mean ± SEM. The data were analyzed using analysis of variance (ANOVA), and the group means were compared by Dunnett’s test. Values were considered statistically significant with P < 0.05. Graph Pad Instat was used for the analysis of data.

 

RESULTS AND DISCUSSION:

Synthetic Chemistry:

The synthesis of target compounds (4A-4J) 2- (2, 3, 4, & 5 substituted phenyl) 3-hydroxy-4H-Chromen-4-one were carried out by reacting 2-hydroxy acetophenone and various aromatic aldehydes in the  presence of potassium hydroxide, methanol and 30% hydrogen peroxide. The synthesized compounds were characterized by IR, NMR, and Mass spectroscopy. The progress of the reaction was monitored by TLC using solvent systems of different polarities. TLC plates are pre-coated silica gel (HF254-200 mesh) aluminum and spots were visualized under U.V chamber and   the proposed structures of the synthesized compounds were ascertained by spectral data.

Table-2: Synthetic flavanoid derivatives

3	R1	R2	R3	R4	4	R1	R2	R3	R4
3A	H	H	-NH2	H	4A	H	H	H	-NH2
3B	H	H	N-(CH3)2	H	4B	H	H	N-(CH3)2	H
3C	OH	H	N-(CH3)2	H	4C	OH	H	N-(CH3)2	H
3D	OH	H	-NH2	H	4D	OH	H	-NH2	H
3E	H	-CH3	OH	H	4E	H	CH3	OH	H
3F	H	-OCH3	-OCH3	H	4F	H	-OCH3	-OCH3	H
3G	H	-OCH3	-OCH3	-OCH3	4G	H	-OCH3	-OCH3	-OCH3
3H	H	-OCH3	OH	-OCH3	4H	H	-OCH3	OH	-OCH3
3I	OH	Cl	H	Cl	4I	H	Cl	Cl	H
3J	OH	H	H	-NH2	4J	OH	H	H	-NH2

 

Table-3: Physicochemical properties of synthesized compounds

Compounds code	M. F	M. W	M. P	Rf -values	Percentage yields
4A	C15H11NO3	253.252	180 C	0.55	68.9%,
4B	C17H15NO3	281.105	175 C	0.45	65.9%,
4C	C17H15NO4	297.1000	181 C	0.47	63.9%,
4D	C15H11NO4	269.068	172 C	0.39	68.5%,
4E	C16H12O4	268.073	181 C	0.45	66.2%,
4F	C17H14O5	298.084	174 C	0.48	62.9%,
4G	C17H14O5	328.09	179 C	0.47	69.9%,
4H	C17H14O6	314.079	177 C	0.46	67.9%,
4I	C16H10Cl2O3	320	175 C	0.44	62.2%,
4J	C15H11NO4	269.06	174 C	0.47	63.6%,

 

Spectral data of the synthesized compounds:

COMPOUND 4A:

FT-IR (KBr) : 3212 cm^-1 (Ar-OH), 1622 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.91 (s, H, -NH₂), 7.01 (s, 1H, OH), 7.00-8.25 (m, 7H, Ar-H),   MS(ESI⁺)m/z : 253.2 (M+H)⁺, HR-MS(ESI⁺)m/z: 253.07 (M+H⁺).

 

COMPOUND 4B: 

FT-IR (KBr) : 3213 cm^-1 (Ar-OH), 1625 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.06 [s, 6H, -N(CH₃)₂], 6.87 (s, 1H, OH), 6.84-8.27 (m, 7H, Ar-H),   MS(ESI⁺)m/z : 281.3 (M+H)⁺, HR-MS(ESI⁺)m/z: 281.105 (M+H⁺).

 

COMPOUND 4C:

FT-IR (KBr) : 3218 cm^-1 (Ar-OH), 1618 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.07 [s, 6H, -N(CH₃)₂], 6.88 (s, 1H, OH), 6.56 (s, 1H, OH),  6.87-8.27 (m, 7H, Ar-H),   MS(ESI⁺)m/z : 297.3  (M+H)⁺, HR-MS(ESI⁺)m/z: 297.1000 (M+H⁺).

COMPOUND 4D: 

FT-IR (KBr) : 3224 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.93 [s, 2H, -NH₂], 6.88 (s, 1H, OH), 6.59 (s, 1H, OH),  6.83-8.22 (m, 7H, Ar-H),   MS(ESI⁺)m/z : 269.2  (M+H)⁺, HR-MS(ESI⁺)m/z: 269.06 (M+H⁺).

 

COMPOUND 4E:

FT-IR (KBr) : 3213 cm^-1 (Ar-OH), 1620 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.92 (s, 3H, -OCH₃), 7.02 (s, 1H, OH), 6.44 (s, 1H, OH),  6.81-8.21 (m, 7H, Ar-H),   MS(ESI⁺)m/z : 268.07  (M+H)⁺, HR-MS(ESI⁺)m/z: 268.26 (M+H⁺).

 

COMPOUND 4F:

FT-IR (KBr) : 3225 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.94 (s, 3H, -OCH₃), 3.96 (s, 3H, -OCH₃), 7.03 (s, 1H, OH),  7.00-8.24 (m, 7H, Ar-H),  MS(ESI⁺)m/z : 298, (M+H)⁺, HR-MS(ESI⁺)m/z: 298.2 (M+H⁺).

 

COMPOUND 4G 

FT-IR (KBr) : 3225 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.94 (s, 3H, -OCH₃), 3.96 (s, 3H, -OCH₃), 3.99 (s, 3H, -OCH₃), 7.03 (s, 1H, OH),  7.00-8.24 (m, 7H, Ar-H),  MS(ESI⁺)m/z : 328, (M+H)⁺, HR-MS(ESI⁺)m/z: 328.3 (M+H⁺).

 

COMPOUND 4H:

FT-IR (KBr) : 3224 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.94 (s, 3H, -OCH₃), 3.99 (s, 3H, -OCH₃), 6.03 (s, 1H, OH), 7.02 (s, 1H, OH), 7.00-8.24 (m, 7H, Ar-H),  MS(ESI⁺)m/z : 314, (M+H)⁺, HR-MS(ESI⁺)m/z: 314.2 (M+H⁺).

COMPOUND 4I:

FT-IR (KBr) : 3224 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 6.03 (s, 1H, OH), 7.02 (s, 1H, OH),    7.00-8.24 (m, 7H, Ar-H),  MS(ESI⁺)m/z : 321.1, (M+H)⁺, HR-MS(ESI⁺)m/z: 320 (M+H⁺).

 

COMPOUND 4J:

FT-IR (KBr) : 3222 cm^-1 (Ar-OH), 1617 cm^-1 ( C=O , Pyrone ring), ¹H-NMR (400MHz, CDCl₃) : δ (ppm) : 3.93 [s, 2H, -NH₂],  6.03 (s, 1H, OH), 7.02 (s, 1H, OH),    7.00-8.24 (m, 7H, Ar-H),  MS(ESI⁺)m/z : 269, (M+H)⁺, HR-MS(ESI⁺)m/z: 269.2 (M+H⁺).

 

Table-4: Structure of synthesized compounds:

Compounds	Structures	Compounds	Structures
4A	2-(3-aminophenyl) – 3 – hydroxyl – 4H-chromen-4-one	4F	2-(3,4-dimethoxyphenyl)-3-hydroxy-4H-chromen-4-one
4B	2-[-4-(dimethylamino)phenyl]-3-hydroxy-4H-chromen-4-one	4G	3-hydroxy-2-(3,4,5-trimethoxyphenyl)-4H-chromen-4-one
4C	2-[4-(dimehylamino)-2-hydroxyphenyl]-3-hydroxy-4H-chromen-4-one	4H	3-hydroxy-2-(4-hydroxy-3,5-dimethoxyphenyl)-4H-chromen-4-one
4D	2-(4-amino-2-hydroxyphenyl)-3-hydroxy-4H-chromen-4-one	4I	2-(3,4-dichlorophenyl)-3-hydroxy-4H-chromen-4-one
4E	3-hydroxy-2-(4-hydroxy-3-methylphenyl-4H-chromen-4-one	4J	2-(5-amino-2-hydroxyphenyl)-3-hydroxy-4H-chromen-4-one

 

 

Experimental Pharmacology:

Acute oral toxicity study:

(i)       Acute oral toxicity studies were performed according to the OECD guideline 423 method.

(ii)     This method has been designed to evaluate the substance at the fixed doses and provide information both for hazard assessment and substance to be ranked for hazard classification purposes.

(iii)    The synthesized compounds were administered initially at a dose of 2000 mg/kg b. w and 1% CMC (p. o) and observed 14 days mortality due to acute toxicity.

(iv)    Careful observation were made at  least thrice a day for the effect on CNS, ANS, motor activity, salivation and other general signs of toxicity were also observed and recorded.

(v)      Since no sign of toxicity observed at 2000 mg/kg b. w. to the group of animals, the LD₅₀ value of the  synthesized compounds expected to exceed 2000 mg/kg b. w. and represented as class 5 (2000 mg/kg < LD50 < 2500 mg/kg).

(vi)    From the toxicity studies the data revealed that all the synthesized compounds proved to be non toxic at tested dose levels and well tolerated by the experimental animals as there LD₅₀ cut of values > 2000 mg/kg b. w.

 

Blood glucose level:

In STZ induced diabetic rats, the blood glucose levels were in the range of 278–280 mg/dL, which were considered as severe diabetes. In the glibenclamide (5 mg/kg) and synthesized compounds (400 mg/kg) treated groups, the peak values of blood sugar significantly on the 21st day, respectively. Hence, in this experimental data displayed that the synthesized compounds significantly decrease the blood glucose level in diabetic rats but values did not return to those of normal controls. Therefore, the compounds possess significant (P < 0.01) ant diabetic activity, when compared with diabetic control. There was significant reduction in blood sugar level (in 21 days) in STZ diabetic animals.

 

Table-5: Effect of synthesized compounds on blood glucose level in STZ induced diabetic rats

Groups		Blood glucose (mg/dL) in day
		0	7	14	21
I		92.2 ± 3.55	94 ± 2.3	94.8 ± 3.05	95.2 ± 2.37
II		278 ± 6.20	281.5 ± 6.01	282.6 ± 5.82	283.4 ± 5.24
III	4A	280.6 ± 5.13ns	257.7 ± 5.94**	190.4 ± 5.25**	116.2 ± 2.27**
	4B	278.6 ± 4.111ns	254.6 ± 5.96**	192.3 ± 5.23**	114.2 ± 2.287**
	4C	279.6 ± 5.12ns	253.2 ± 5.93**	194.2 ± 5.22**	116.2 ± 2.24**
	4D	275.6 ± 5.14ns	257.6 ± 5.96**	191.5 ± 5.21**	115.2 ± 2.23**
	4E	282.5 ± 5.15ns	249.2 ± 5.88**	195.1 ± 5.26**	116.1 ± 2.22**
	4F	277.3 ± 5.15ns	250.4 ± 5.89**	189.6 ± 5.16**	115.1 ± 2.21**
	4G	276.4 ± 5.12ns	257.6 ± 5.99**	194.3 ± 5.18**	114.2 ± 2.25**
	4H	281.9± 5.11ns	253.2± 5.96**	189.2 ± 5.20**	118.2 ± 2.26**
	4I	283.1 ± 5.45ns	251.7 ± 5.92**	190.1 ± 5.15**	117.2 ± 2.25**
	4J	285.8 ± 5.24ns	256.6 ± 5.55**	192.4 ± 5.22**	119.2 ± 2.27**
IV		282.2 ± 6.88 ns	247.8 ± 5.76**	178.4 ± 6.49**	116.6 ± 3.19**

Values are mean ± SEM; n = 5 in each group except in diabetic control group where n = 4 because one animal died on the 8th day. Ns P > 0.05 (non-significant), **P < 0.01 (highly significant) when compared to diabetic control rats; Compounds or glibenclamide was administered daily for 21 days. For glucose estimation, blood was collected just before the drug administration on the 0 day and 1 h after the drug administration on the 7th, 14th day and 21st day.

 

Fig 1: The effect of synthesized compounds and glibenclamide on blood glucose level in STZ induced diabetic rats at various days (on 0 day, 7th day, 14th day, and 21st day).

Serum lipid profile:

The serum profile lipids also determined by analysis of serum sample withdrawn from the all groups. The untreated diabetic rats’ serum levels of cholesterol and TG were significantly increased which was significantly declined with administration of synthesized compounds. The results obtained from the glibenclamied on serum TG and cholesterol in the diabetic rats was comparable to those of synthesized compounds. Total cholesterol and TG were significantly increased in diabetic group in comparison to control group. Administration of synthesized compounds for 21 days potentially decreased the serum levels of cholesterol and TG in comparison to diabetic control rats.

 

Table-6: Effect of synthesized compounds on serum lipid profile in STZ-induced diabetic rats

Groups	Cholesterol (mg/dL)	Triglycerides (mg/dL)
I	80.12 ± 2.59	68.3 ±4.15
II	129 ± 9.39	119.5 ± 4.94
III (4A-4J)	101.3 ± 4.03*-103.5±5.01*	93.9 ± 2.94*-93.12±2.99*
IV	86.18± 2.11**	82.12 ± 5.43**

Values are mean ± SEM; n = 5 in each group except in diabetic control group where n = 4 because one animal died on the 8th day. *P < 0.05 (significant),**P < 0.01 (highly significant) when compared to diabetic control rats.

 

Determination of urinary glucose and Ketone bodies:

Urine analysis on day 0 displayed the presence of glucose (+++) and ketone (trace) in a all groups, except normal control and on 21st day glucose and ketone traces were absent in synthesized compounds and glibenclamide treated groups where as urinary glucose and ketone bodies were present in diabetic control group.

 

Table-7: Effect of synthesized compounds on urinary glucose and ketone bodies in STZ-induced diabetic rats

Groups	0 day		21st day
	Glucose	Ketone bodies	Glucose	Ketone bodies
I	−	−	−	−
II	+++	Trace	+++	Trace
III (4A-4J)	+++	Trace	−	−
IV	+++	Trace	−	−

- = absence of glucose, +++ = 1.1 g/dL, - = absence of ketone, Trace = 4.98 mg/dL

 

CONCLUSION:

From the present experimental data, here we concluded that all the synthesized compounds possessed potential anti diabetic activity against streptozocin induced rats which was proved by assessment of bioanalytes from serum and urine sample.

 

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Received on 26.08.2018         Modified on 11.09.2018

Accepted on 07.10.2018         © AJRC All right reserved