The Ir Spectra Of The Compounds Biology Essay

The thaw points were taken in unfastened capillary tubing method. The IR spectra of the compounds were record on ABB Bomem FT-IR spectrometer MB 104 IR spectra recorded with K bromide pellets. The 1H-NMR spectra of synthesized compounds were recorded on instrument BRUKER NMR spectrometer in DMSO. The Mass spectra of synthesized compound were recorded JEOL GCmate. TLC method was used to look into pureness of compounds. TLC home bases are Pre -coated Silica gel ( HF254-200 mesh ) aluminium home bases utilizing ethyl ethanoate: n-hexane was used as eluent and visualized under UV- chamber. The IR, 1H-NMR and MASS spectra were consistent with assigned construction of synthesized compounds.

4.2. SYNTHETIC Scheme

4.3. SYNTHESIS STEPS OF TARGET COMPOUNDS

Measure: 1

Measure: 2

Measure: 3

Title compounds ( G1-G12 )

Measure: 1

Ethyl -4-acetamido phenoxy acetate61

A mixture of p-acetamido phenol ( 0.01 mol ) and ethyl chloroacetate ( 0.01 mol ) was refluxed by utilizing dry propanone in presence of anhydrous K carbonate ( K2CO3 ) for 6hrs. The reaction mixture was cooled and so poured in to oppress ice. The solid merchandise obtained, these merchandise was filtered, dried and recrystallized utilizing ethyl alcohol.

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Measure: 2

4-Acetamido phenoxy ethanoyl group hydrazide

The ethyl-4-acetamido phenoxy ethanoate ( 0.01 mol ) , hydrazine hydrate ( 0.01 mol ) was refluxed in the presence of ethyl alcohol for 5 hour. The reaction mixture was cooled and so poured in to oppress ice. The solid merchandise obtained, these merchandise was filtered, dried and recrystallized utilizing ethyl alcohol.

Measure: 3

2- ( 4-Acetamido phenoxy methyl ) -5-aryl substituted – 1, 3, 4-oxadiazole62

A mixture of 4-Acetamido phenoxy ethanoyl group hydrazide ( 0.01 mol ) and assorted aromatic acids ( 0.01 mol ) in P oxychloride ( 10 milliliter ) was exposed to microwaves at 350 W. intermittently at 2min. intervals for 8-15 min. Completion of the reaction procedure, monitored by TLC home bases. The contents were cooled and poured into the crushed ice. It was neutralized with Na hydrogen carbonate solution and the solid merchandise obtained, these merchandise was filtered, dried and recrystallized utilizing ethyl alcohol.

4.3. PHYSICO-CHEMICAL PROPERTIES AND SPECTRAL DATA OF SYNTHESIZED COMPOUNDS

Compound

G1

IUPAC NAME

4- ( ( 5- ( 3-aminophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H16N4O3

Molecular weight

324.33

Melting point ( 0C )

1160C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2: 3

Rf value

0.77

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

54 %

IR ( KBr ) I? ( cm-1 )

3393.16 cm-1 ( Ar-NH ) , 1633.67 cm-1 ( C=N ) , 1575.88 cm-1 ( C=C ) , 1069.05 cm-1 ( -C-O-C- ) , 3132.54 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.45-7.4 ( s, 8H, Ar-H ) , 5.17 ( s, 2H, -CH2 ) , 4.1 ( s, 2H, -NH2 ) , 2.05 ( s,1H, -CH3 ) , 8.05 ( s, 1H, -NH ) .

Mass ( m/e value )

% comparative copiousness

324.03 ( M+ ) ( 5.1 ) , 310.87 ( 4 ) , 296.22 ( 8.25 ) , 282.76 ( 2.2 ) , 272.38 ( 2.32 ) , 262.6432 ( 7.3 ) , 248.34 ( 11 ) , 217.12 ( 15 ) , 207.14 ( 7 ) , 116.67 ( 18 ) , 58.33 ( B )

Indian Institute of Technology, Chennai

File: G1 Date Run: 12-20-2010 ( Time Run: 10:13:54 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G2

IUPAC NAME

4- ( ( 5- ( 2-aminophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H16N4O3

Molecular weight

324.33

Melting point ( 0C )

1230C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2: 3

Rf value

0.76

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

56 %

IR ( KBr ) I? ( cm-1 )

3381.92 cm-1 ( Ar-NH ) , 1677.76 cm-1 ( C=N ) , 1530.51 cm-1 ( C=C ) , 1089.81 cm-1 ( -C-O-C- ) , 3118.42 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.46-7.45 ( s,8H, Ar-H ) , 5.21 ( s, 2H, -CH2 ) , 4.05 ( s, 2H, -NH2 ) ,2.12 ( s, 1H, -CH3 ) , 7.95 ( s, 1H, -NH ) ,

Mass ( m/e value )

% comparative copiousness

324.57 ( M ) ( 6 ) , 284.34 ( 12.3 ) ,262.64 ( 7.8 ) , 284.34 ( 10.1 ) , 216.62 ( 22.3 ) , 174.45 ( 8 ) , 156.98 ( 7 ) , 80.10 ( B )

Indian Institute of Technology, Chennai

File: G2 Date Run: 12-20-2010 ( Time Run: 10:20:34 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G3

IUPAC NAME

4- ( ( 5- ( 2-chloro-5-nitrophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H13ClN4O5

Molecular weight

388.76

Melting point ( 0C )

1350C

TLC dissolver ratio

Ethyl ethanoate: Hexane

3: 2

Rf value

0.64

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

60 %

IR ( KBr ) I? ( cm-1 )

3381.95 cm-1 ( Ar-NH ) , 1684.44 cm-1 ( C=N ) , 1586.2 cm-1 ( C=C ) , 1064.25 cm-1 ( -C-O-C- ) , 1365.57 cm-1 ( N=O ) , 801.35 cm-1 ( C-Cl ) , 3130.43 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.74-8.36 ( m, 7H, Ar-CH ) ,5.31 ( s, 2H, -CH2 ) ,2.31 ( s, 1H, -CH3 ) , 8.16 ( s, 1H, -NH ) ,

Mass ( m/e value )

% comparative copiousness

390.26 ( M ) ( 4 ) , 388.71 ( 8.1 ) , 362.27 ( 4.2 ) , 233.28 ( 5 ) , 217.31 ( 8.9 ) , 182.52 ( 5 ) , 96.79 ( 7 ) , 78.82 ( B )

Indian Institute of Technology, Chennai

File: G3 Date Run: 12-20-2010 ( Time Run: 10:26:25 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G4

IUPAC NAME

4- ( ( 5- ( 4-chloro-3-nitrophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H13ClN4O5

Molecular weight

388.76

Melting point ( 0C )

2030C

TLC dissolver ratio

Ethyl ethanoate: Hexane

1:1

Rf value

0.68

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

62 %

IR ( KBr ) I? ( cm-1 )

3382.02 cm-1 ( Ar-NH ) , 1677.79 cm-1 ( C=N ) , 1530.6 cm-1 ( C=C ) , 1089.68 cm-1 ( -C-O-C- ) , 1372.45 cm-1 ( N=O ) , 813.038 cm-1 ( C-Cl ) , 3117.5 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.83-8.42 ( m, 8H, Ar-CH ) , 5.35 ( s, 2H, -CH2 ) , 2.07 ( s, 1H, – CH3 ) , 8.24 ( s, 1H, -NH ) ,

Mass ( m/e value )

% comparative copiousness

390.27 ( M+ ) ( 5 ) , 388.76 ( 13 ) , 380.25 ( 8 ) , 261.63 ( 8 ) , 182.52 ( 5 ) , 167.62 ( 17 ) , 156.56 ( 19 ) , 81.97 ( B ) .

Indian Institute of Technology, Chennai

File: G4 Date Run: 12-20-2010 ( Time Run: 10:32:48 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G5

IUPAC NAME

4- ( ( 5- ( 2-fluorophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H14FN3O3

Molecular weight

327.31

Melting point ( 0C )

1870C

TLC dissolver ratio

Ethyl ethanoate: Hexane

1:1

Rf value

0.75

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

64 %

IR ( KBr ) I? ( cm-1 )

3392.09 cm-1 ( Ar-NH ) , 1617.53 cm-1 ( C=N ) , 1528.16 cm-1 ( C=C ) , 1093.52 cm-1 ( -C-O-C- ) , 1371.78 cm-1 ( C-F ) , 3114.61 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

2.21 ( s, 1H, -CH3 ) , 8.09 ( s, 1H, -NH ) , 5.21 ( s, 1H, -CH2 ) , 6.7-8.01 ( m, 8H, Ar-CH ) .

Mass ( m/e value )

% comparative copiousness

327.37 ( M+ ) ( 6.3 ) , 310.37 ( 2.3 ) , 299.57 ( 3 ) , 282.87 ( 3.9 ) , 266.22 ( 5 ) , 249.61 ( 1.2 ) , 232.72 ( 4 ) , 104.86 ( 8.1 ) , 75.50 ( B )

Indian Institute of Technology, Chennai

File: G5 Date Run: 12-20-2010 ( Time Run: 10:40:51 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G6

IUPAC NAME

4- ( ( 5- ( 4-methoxyphenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C18H17N3O4

Molecular weight

353.35

Melting point ( 0C )

1740C

TLC dissolver ratio

Ethyl ethanoate: Hexane

3:4

Rf value

0.76

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

56 %

IR ( KBr ) I? ( cm-1 )

3457.55 cm-1 ( Ar-NH ) , 1625.05 cm-1 ( C=N ) , 1585.42 cm-1 ( C=C ) , 1094.6 cm-1 ( -C-O-C- ) , 3123.84 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.8-7.56 ( m, 8H, Ar-CH ) , 2.22 ( s, 1H, -CH3 ) ,3.70 ( s, 1H, , -CH3 ) , 5.21 ( s, 2H, -CH2 ) ,8.14 ( s, 1H, NH )

Mass ( m/e value )

% comparative copiousness

339.84 ( M+ ) ( 6 ) , 324.91 ( 2.2 ) , 307.71 ( 3.9 ) , 296.22 ( 1.5 ) , 249.64 ( 16 ) , 160.35 ( 17 ) ,132.55 ( 24 ) , 116.67 ( 18 ) , 87.74 ( B ) .

Indian Institute of Technology, Chennai

File: G6 Date Run: 12-20-2010 ( Time Run: 10:46:27 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G7

IUPAC NAME

4- ( ( 5- ( 4-ethoxyphenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C19H19N3O4

Molecular weight

353.37

Melting point ( 0C )

1750C

TLC dissolver ratio

Ethyl ethanoate: Hexane

1:1

Rf value

0.73

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

64 %

IR ( KBr ) I? ( cm-1 )

3430.18 cm-1 ( Ar-NH ) , 1630.28 cm-1 ( C=N ) , 1589.85 cm-1 ( C=C ) , 1091.04 cm-1 ( -C-O-C- ) , 3123.31 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.74-7.37 ( m, 8H, Ar-CH ) , 5.24 ( s, 2H, -CH2 ) , 2.25 ( s, 1H, -CH3 ) , 8.07 ( s, 1H, -NH ) ,1.31 ( s,1H, -CH3 ) ,

Mass ( m/e value )

% comparative copiousness

353.16 ( M+ ) ( 4.9 ) , 335.13 ( 3.9 ) , 325.22 ( 2 ) , 302.39 ( 2.7 ) , 249.58 ( 6.8 ) , 204.96 ( 4.9 ) , 183.26 ( 5 ) ,160.32 ( 5.8 ) , 115.64 ( 7.1 ) , 91.31 ( B )

Indian Institute of Technology, Chennai

File: G7 Date Run: 12-20-2010 ( Time Run: 10:52:34 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G8

IUPAC NAME

4- ( ( 5-o-tolyl-1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C18H17N3O3

Molecular weight

323.35

Melting point ( 0C )

2110C

TLC dissolver ratio

Ethyl ethanoate: Hexane

1:1

Rf value

0.65

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

63 %

IR ( KBr ) I? ( cm-1 )

3354.23 cm-1 ( Ar-NH ) , 1632.99 cm-1 ( C=N ) , 1584.68 cm-1 ( C=C ) , 1092.53 cm-1 ( -C-O-C- ) , 3123.84 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

7.41 ( m, 4H, Ar-CH ) , 2.31 ( s, 1H, -CH3 ) , 8.16 ( s, 1H, -NH ) , 5.31 ( s, 1H, -CH2 ) ,

Mass ( m/e value )

% comparative copiousness

323.38 ( M+ ) ( 8 ) , 277.92 ( 7 ) , 266.92 ( 6.2 ) , 249.72 ( 1.1 ) , 232.7228 ( 11.8 ) , 158.60 ( 16.0 ) , 140.88 ( 14 ) , 132.84 ( 40 ) , 80.40 ( B )

Indian Institute of Technology, Chennai

File: G8 Date Run: 12-20-2010 ( Time Run: 10:59:56 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G9

IUPAC NAME

4- ( ( 5- ( 3-nitrophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H14N4O5

Molecular weight

354.32

Melting point ( 0C )

1690C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2:2

Rf value

0.72

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

64 %

IR ( KBr ) I? ( cm-1 )

3382.43 cm-1 ( Ar-NH ) , 1703.01 cm-1 ( C=N ) , 1592.32 cm-1 ( C=C ) , 1088.54 cm-1 ( -C-O-C- ) , 1378.11 cm-1 ( N=O ) , 3112.69 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.41-7.8 ( m, 8H, Ar-CH ) , 2.42 ( s, 3H, -CH3 ) , 8.13 ( s, 1H, -NH ) , 5.21 ( s, 2H, CH2 ) .

Mass ( m/e value )

% comparative copiousness

354.16 ( M ) ( 3.8 ) , 335.16 ( 4.8 ) , 302.39 ( 3.1 ) , 287.43 ( 3.7 ) , 249.58 ( 7.1 ) , 226.00 ( 5.8 ) , 204.96 ( 6.7 ) , 127.56 ( 13.1 ) , 103.69 ( 9 ) , 89.93 ( B ) .

Indian Institute of Technology, Chennai

File: G9 Date Run: 12-20-2010 ( Time Run: 11:07:26 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G10

IUPAC NAME

4- ( ( 5- ( 2,6-difluorophenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H13F2N3O3

Molecular weight

345.3

Melting point ( 0C )

1800C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2:3

Rf value

0.74

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

53 %

IR ( KBr ) I? ( cm-1 )

3368.58 cm-1 ( Ar-NH ) , 1673.42 cm-1 ( C=N ) , 1545.03 cm-1 ( C=C ) , 1085.04 cm-1 ( -C-O-C- ) , 1378.04 cm-1 ( C-F ) , 3115.62 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.6-7.82 ( s, 8H, Ar-CH ) , 2.5 ( s, 3H, -CH3 ) , 8.03 ( s, 1H, -NH ) , 5.22 ( s, 2H, -CH2 ) ,

Mass ( m/e value )

% comparative copiousness

345.13 ( M+ ) ( 2.8 ) , 333.16 ( 1.5 ) , 325.42 ( 2.7 ) , 286.43 ( 2.6 ) , 183.26 ( 6 ) , 160.62 ( 7 ) , 140.65 ( 16 ) , 115.64 ( 33 ) , 95.53 ( B )

Indian Institute of Technology, Chennai

File: 10 Date Run: 12-20-2010 ( Time Run: 11:16:13 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G11

IUPAC NAME

4- ( ( 5- ( 2-hydroxyphenyl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C17H15N3O4

Molecular weight

325.32

Melting point ( 0C )

2150C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2:3

Rf value

0.72

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

68 %

IR ( KBr ) I? ( cm-1 )

3382.83 cm-1 ( Ar-NH ) , 1654.42 cm-1 ( C=N ) , 1541.89. cm-1 ( C=C ) , 1090.01 cm-1 ( -C-O-C- ) , 3118.84 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

6.7-7.6 ( s, 6H, Ar-CH ) , 2.11 ( s, H, -CH3 ) , 8.00 ( s, 1H, -NH ) , 5.12 ( s, 1H, -CH2 )

Mass ( m/e value )

% comparative copiousness

325.42 ( M ) ( 11.1 ) , 318.68 ( 16 ) , 292.76 ( 7 ) , 276.89 ( 20 ) , 249.99 ( 8.2 ) , 236.0277 ( 28.1 ) , 203.2266 ( 76 ) , 182.2587 ( 8 ) , 134.4966 ( 32 ) , 116.55 ( B )

Indian Institute of Technology, Chennai

File: 11 Date Run: 11-20-2010 ( Time Run: 11:20:34 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

Compound

G12

IUPAC NAME

4- ( ( 5- ( pyridin-3-yl ) -1,3,4-oxadiazol-2-yl ) methoxy ) phenyl ) ethanamide

Molecular expression

C16H14N4O3

Molecular weight

310.31

Melting point ( 0C )

2100C

TLC dissolver ratio

Ethyl ethanoate: Hexane

2: 3

Rf value

0.71

Solubility

Insoluble- Water

Slightly soluble- Chloroform, Ethanol

Freely soluble- DMF, DMSO

Output ( % )

62 %

IR ( KBr ) I? ( cm-1 )

3482.34 cm-1 ( Ar-NH ) , 1654.26 cm-1 ( C=N ) , 1528.16. cm-1 ( C=C ) , 1093.52 cm-1 ( -C-O-C- ) , 3138.86 cm-1 ( Ar-CH )

1H-NMR I? ( ppm )

7.31-7.82 ( s, 8H, Ar-CH ) , 2.1 ( s, 1H, -CH3 ) , 8.13 ( s, 1H, -NH ) , 5.2 ( s, 1H, -CH2 )

Mass ( m/e value )

% comparative copiousness

310.36 ( M+ ) ( 2.8 ) , 289.96 ( 0.9 ) , 248.58 ( 1.5 ) , 231.61 ( 0.8 ) , 157.07 ( 37 ) , 131.35 ( 62.1 ) , 115.46 ( 3.8 ) , 103.53 ( 62.2 ) , 96.13 ( B )

Indian Institute of Technology, Chennai

File: 12 Date Run: 12-20-2010 ( Time Run: 11:27:44 )

Sample:

Instrument: JEOL GCmate

Inlet: Direct Probe Ionization manner: EI+

CHAPTER-5

PHARMACOLOGICAL EVALUATION

Material and Method

The Assorted animate beings and stuffs used for present survey and at that place resources as follows,

Wister rats

1 % Carboxy Methyl Cellulose ( Rolax Laboratory )

Pentazocin ( Abbott )

Diclofenac Na ( Rolax Laboratory )

Water for injection ( Global Pharma )

Carrageenan ( Sigma )

Synthesized compounds ( G1- G12 )

Plethysmograph ( Indigenous )

The animate beings used in present survey such as Wister rats weighing 150-200 gram were kept in settlement coops at 28 i‚± 20C, comparative humidness of 50-55 % under 12hrs visible radiation and dark rhythm. All animate beings were feed with standard animate being provender, H2O and labium. The trial compounds were administered orally, in the signifier of suspension utilizing 1 % CMC as suspending agent. The experimental dosage was selected between minimal effectual dosage and maximal non lethal non lethal dosage. All carnal experimentation was performed harmonizing to protocols and recommendation of carnal moralss commission.

IAEC Reference No: IAEC/XXXI/09/CLBMCP dated 22/09/2010.

ANOVA followed by dunnet ‘s trial was performed to determine the significance of exhibited analgetic and anti-inflammatory activities of synthesized compounds.

Following activities were carried out to the synthesized compounds by holla mentioned methods.

Acute unwritten toxicity ( acute toxic category method in mice )

Analgesic activity ( Tail submergence Method )

Anti-inflammatory activity ( carrageenin induced acute paw hydrops in rats )

5.1 EVALUATION OF ACUTE ORAL TOXICITY

Introduction

Acute unwritten toxicity defines to those inauspicious effects happening following unwritten disposal of a individual dosage of a substances or multiple dose given within 24 h. The assorted methods used to measure the acute unwritten toxicity as follows.

Fixed dose process ( OECD guideline-420 )

Acute toxic category method ( OECD guideline -423 )

Up and down process ( OECD guideline -425 )

OECD guideline -42363

a ) OECD Guidelines for the Testing of Chemicals are sporadically reviewed in the visible radiation of scientific advancement or altering assessment patterns. The original Guideline 423 was adopted in March 1996 as the 2nd option to the conventional acute toxicity trial, described in Test Guideline 401.

I ) International understanding has been reached on consonant LD50 cut-off values for the categorization of chemical substances, which differ from the cut-offs recommended in the 1996 version of the Guideline. two ) Testing in one sex ( normally females ) is now considered sufficient.

B ) The acute toxic category method set out in this Guideline is a bit-by-bit process with the usage of 3 animate beings of a individual sex per measure. Depending on the mortality and/or the moribund position of the animate beings, on mean 2-4 stairss may be necessary to let judgement on the acute toxicity of the trial substance. This process is consistent, uses really few animate beings and is able to rank substances in a similar mode to the other acute toxicity proving methods. ( Test Guidelines 420 and 425 ) . The acute toxic category method is based on biometric ratings with fixed doses, adequately separated to enable a substance to be ranked for categorization intents and hazard appraisal. The method as adopted in 1996 was extensively validated in vivo against LD50 informations obtained from the literature, both nationally and internationally.

degree Celsius ) Guidance on the choice of the most appropriate trial method for a given intent can be found in the Guidance Document on Acute Oral Toxicity Testing.

This Guidance Document besides contains extra information on the behavior and reading of Test Guideline 423.

Experimental Protocol ( acute toxic category method in rats )

In the present survey acute unwritten toxicity of the synthesized compounds were performed by acute toxic category method. In this method the toxicity of synthesized compounds were tested utilizing a measure wise process, each measure utilizing three mice of individual sex ( female ) . The mice were fasted prior to dosing ( nutrient but H2O should be with held ) for three to four hours. Following the period of fasting the animate being should be weighted and synthesized compound were administered orally at a dose 2000 mgiˆ?kg organic structure weight. Animals were observed separately after dosing at least one time during the first 30min ; sporadically during the first 24 H with particular attending giving during the first 4 H and day-to-day thereafter, for sum of 14 yearss. As know mortality observed with the above dosage. Test compound dosage reduced by specific intervals. The mortality non observed at the dose 1000 mgiˆ?Kg. So a series of doses 100 and 200 mgiˆ?Kg organic structure weight were selected for their pharmacological rating. The trial process with the get downing dosage of 2000 mgiˆ?Kg organic structure weight as per OECD-423 guidelines was shown as follows.

Fig.1

Flow chart for acute toxic category method ( OECD guideline 423 ) get downing dosage of 2000 mgiˆ? Kg organic structure weight p.o

Table: 5.1

Dose Selection by Acute toxicity category method ( OECD ) guide lines 423 of European union

Sr. No.

Treatment group

Dose mg/kg

Sign of toxicity

Onset of toxicity

Duration

1

G1

200

400

No

No

14 yearss

2

G2

200

400

No

No

14 yearss

3

G3

200

400

No

No

14 yearss

4

G4

200

400

No

No

14 yearss

5

G5

200

400

No

No

14 yearss

6

G6

200

400

No

No

14 yearss

7

G7

200

400

No

No

14 yearss

8

G8

200

400

No

No

14 yearss

9

G9

200

400

No

No

14 yearss

10

G10

200

400

No

No

14 yearss

11

G11

200

400

No

No

14 yearss

12

G12

200

400

No

No

14 yearss

Consequences and treatment

Acute unwritten toxicity surveies were performed harmonizing to the OECD guideline 423 method.

This method has been designed to measure the substance at the fixed doses and supply information both for jeopardy appraisal and substance to be ranked for hazard categorization intents.

The synthesized compounds were administered ab initio at a dosage of 2000mg/kg b.w and 1 % CMC ( p.o ) and observed 14 yearss mortality due to acute toxicity.

Careful observation were made at least thrice a twenty-four hours for the consequence on CNS, ANS, motor activity, salivation and other general marks of toxicity were besides observed and recorded.

Since no mark of toxicity observed at 2000 mg/kg b.w. to the group of animate beings, the LD50 value of the rubric compounds ( G1-G12 ) expected to transcend 2000 mg/kg b. w. and represented as category 5 ( 2000 mg/kg & lt ; LD50 & lt ; 2500 mg/kg )

From the toxicity surveies the informations revealed that all the synthesized compounds proved to be non toxic at tested dosage degrees and good tolerated by the experimental animate beings as there LD50 cut of values & gt ; 2000 mg/kg b. tungsten.

5.2 EVALUATION OF ANTI-INFLAMMATORY ACTIVITY

Introduction

Anti-inflammatory agents are substance which modifies the inflammatory reaction. The chief anti-inflammatory agents are the glucocorticoids and the non steroidal anti-inflammatory drugs ( NSAIDS )

Evaluation method

The three of import facets of redness that render themselves readily to measuring are erythema, hydrops and formation of granulation tissue. Compounds claimed to posses anti-inflammatory activity can be evaluated either by their ability to cut down one one or more of these phenomena in by experimentation induced redness or by proving their anti-inflammatory activity in experimental arthritis produced in animate beings. The normally employed methods are,

Erythematic check

Granuloma check

Edema assay

Experimental arthritis check

Assorted

Erythema assay

In this method irradiation of shaven back tegument of guinea hog with UV visible radiation cause erythema which can be reduced by anti-inflammatory agents.

Granuloma check

There are two types of granuloma checks such as cotton wool pellet and granuloma pouch method.

Edema assay

The hydrops can be produced in experimental animate beings by the local injection of substance like methanal, carrageenin, histamine, dextrin egg white

Experimental arthritis check

Poly arthritis induced in rats by injection of dead tubercle B suspended in liquid paraffin is often used method china clay ; talc and even quicksilvers have besides been injected straight into articulations of rats and pigeons to bring on arthritis.

Assorted

Localized inflammatory reaction can be produced in rats by

Intra pleural injection of gum terpentine

Intra peritoneal injection of methanal

Experimental Procedure: –

Carrageenan Induced Paw Oedema Method in Rats64

Anti-inflammatory activity was performed by carrageenin induced paw hydrops method in rats. Diclofenac Na ( 20 mg/kg i.p ) was administered as standard drug for comparing. The synthesized compounds were administered at two dose degree ( 200 and 400 mg/kg ) .orally 30 proceedingss prior to the disposal of 0.1ml/kg organic structure weight of carrageenin used in saline ( 1 % w/v ) into the sidelong malleolus of sub-planter part of the rats left back paw. The paw volumes were measured utilizing the quicksilver supplanting technique with the aid of a plethysmonograph instantly before and 30 proceedingss, 1, 2 and 3 hr after carrageenin injection. The per centum suppression of paw hydrops was calculated by utilizing the undermentioned expression.

Percentage protection = [ ( control-test ) /control ] A-100

Table 5.2.1

Anti Inflammatory Activity of the Synthesized Compounds ( 200 mg/kg )

Compounds

30 min

60 min

120 min

180 min

MEAN +SEM

%

MEAN +SEM

%

MEAN +SEM

%

MEAN +SEM

%

G1

0.70A± 0.13

9.09

0.77A±0.02NS

18.94

0.96A±0.09

35.57

0.85A±0.11

23.42

G2

0.65A±0.05*

15.68

0.68A±0.04

28.43

0.82A±0.11**

44.96

0.79A±0.13*

28.82

G3

0.62 A± 0.06

19.48

0.74A±0.05

22.10

0.88A±0.32

40.93

0.62A±0.23*

44.14

G4

0.58A± 0.04

24.67

0.64A±0.11*

32.63

0.76A±0.14**

50.33

0.63A±0.12

43.24

G5

0.70 A±0.08

9.09

0.78A±0.07

17.89

0.95A±0.34

36.24

0.89A±0.04

19.81

G6

0.70A± 0.05

9.09

0.77A±0.05

18.94

0.80A±0.13*

46.30

0.64A±0.06*

42.34

G7

0.56A± 0.09*

27.27

0.68A±0.06*

28.42

0.77A±0.09*

48.32

0.65A±0.11*

41.44

G8

0.75A± 0.05

2.59

0.82A±0.12

13.68

0.92A±0.11

38.25

0.76A±0.21

29.72

G9

0.59A±0.11*

23.37

0.78A±0.03

17.89

0.90A±0.05

39.59

0.68A±0.09

38.73

G10

0.74A± 0.09

3.89

0.85A±0.07

10.5

0.98A±0.14

34.22

0.79A±0.01

28.82

G11

0.64A± 0.05

16.88

0.69A±0.7*

27.36

0.84A±0.21*

43.62

0.74A±0.15

33.33

G12

0.63A± 0.05

18.18

0.71A±0.05

25.26

0.81A±0.04NS

45.63

0.75A±0.11

32.43

Diclofenac Sodium

0.56A±0.09**

0.62A±0.13**

0.70A±0.17**

0.58A±0.05**

Significant differences with regard to command was evaluated by ( ANOVA ) , Dunnet ‘s t trial * P & lt ; 0.05, **P & lt ; 0.01, NS ( Non important ) , % ( Percentage decrease of hydrops ) .

Table 5.2.2

Anti Inflammatory Activity of the Synthesized Compounds ( 400 mg/kg )

Compounds

30 min

60 min

120 min

180 min

MEAN +SEM

%

MEAN +SEM

%

MEAN +SEM

%

MEAN +SEM

%

G1

0.587A± 0.12

40.40

0.411A±0.02NS

39.45

0.586A±0.11

35.60

0.451A±0.09

28.66

G2

0.553A±0.04*

43.85

0.407A±0.04

40.14

0.566A±0.13*

37.80

0.442A±0.11**

29.13

G3

0.502 A± 0.06

49.03

0.355A±0.05

47.79

0.502A±0.23*

44.33

0.412A±0.32

33.33

G4

0.501A± 0.04

49.13

0.341A±0.11*

49.85

0.498A±0.12

45.27

0.404A±0.14**

34.30

G5

0.530 A±0.08

46.19

0.385A±0.07

43.38

0.530A±0.04

41.09

0.428A±0.34

30.40

G6

0.549A± 0.05

44.26

0.392A±0.05

42.35

0.593A±0.06*

37.83

0.435A±0.13*

29.26

G7

0.592A± 0.04*

39.89

0.426A±0.06*

37.35

0.608A±0.11*

33.18

0.455A±0.09*

26.01

G8

0.666A± 0.05

32.38

0.444A±0.12

34.70

0.643A±0.21

31.53

0.478A±0.11

22.27

G9

0.696A±0.15*

29.34

0.457A±0.03

32.79

0.629A±0.09

30.87

0.473A±0.05

23.08

G10

0.525A± 0.09

46.70

0.514A±0.07

45.00

0.526A±0.01

42.19

0.422A±0.14

31.38

G11

0.493A± 0.05

49.74

0.529A±0.7*

51.61

0.492A±0.15

45.73

0.402A±0.21*

34.63

G12

0.489A± 0.1

50.35

0.504A±0.05

55.29

0.469A±0.11

48.46

0.392A±0.04NS

36.37

Diclofenac Sodium

0.498A±0.09**

0.319A±0.13**

0.480A±0.05**

0.397A±0.17**

Significant differences with regard to command was evaluated by ( ANOVA ) , Dunnet ‘s t trial * P & lt ; 0.05, **P & lt ; 0.01, NS ( Non important ) , % ( Percentage decrease of hydrops ) .

RESULTS AND DISCUSSION

Anti-inflammatory activity of the synthesized compounds was evaluated by carrageenin induced paw hydrops method. The activity was studied at 200 and 400 mg/kg b.w. p.o and their effects were measured at 30, 60, 120 and 180 min.

From the informations shown in tabular arraies, the undermentioned observations we made:

Mild to good anti-inflammatory activity was observed for all the synthesized compounds. Graded dose response was besides observed.

All compounds exhibited highest activity at 120 min.

When compared with standard drug diclofenac Na ( 20 mg/kg i.p ) G1, G2, G7 and G8 exhibited equipotent anti-inflammatory activity.

Compounds G2, G4, G7 and G9 exhibited moderate anti-inflammatory activity.

These compound shows anti-inflammatory activity because these compounds holding NH2, NO2, Cl groups.

5.3 EVALUATION OF ANALGESIC ACTIVITY

Introduction

Definition:

“ Analgesics are agents which relieve or decrease pain esthesis by increasing threshold to painful stimulations without doing loss of consciousness ” .

Pain agencies of many diseases necessitating intervention with anodynes. The dependence liability of opioids leads to intensive research for compounds without side consequence.

Many attacks have been used to distinguish the assorted actions of strong anodynes by developing carnal theoretical accounts non merely for analgetic activity. Analgesic effects in animate beings are comparable with the curative effects in adult male. Painful stimulations can dwell of direct stimulation of the motorial centripetal nervousnesss or stimulation of hurting receptors by assorted agencies such as heat or force per unit area. The function of endogenous peptides such as endorphins gives more insight into encephalon procedures and the action of cardinal anodynes.

Mechanism of action

Cellular action-

Action site is protein coupled Opioid receptors. The suppression of adenocyclase.

Reducing the intracellular camp content impacting protein phosphorylation Pathways.

Exert effects on ion channels through a direct G-protein yoke to the channel. Promote the gap of K channels and suppress the gap of voltage-gated Ca channels.

These membrane effects cut down both neural irritability ( because of the increasing K+ conductance causes hyper polarisation of the membrane ) and transmitter release ( due to suppression of the Ca2+ entry ) .

The overall consequence is hence repressive at the cellular degree. Increase activity in some neural tracts by stamp downing the fire of repressive interneuron.

The other analgetic mechanism green goodss by consequence on nociceptive tract.

Evaluation methods

The assorted methods used to measure anticonvulsant activity are as follows,

Hoppners tail cartridge holder method

Hot home base method

Beaming heat method

Tail submergence method

Grid daze trial

Electric stimulation method

Formalin trial in rats

Experimental protocol ( Tail submergence method ) 65

The analgetic activity was determined by popular method like tail-immersion method. Wister rat ( n=6 ) of either sex selected by random trying technique was used for the survey. Pentazocine at the dosage of 20 mg/kg ( i.p. ) was administered as standard drug for comparing. The trial compounds at 2 dosage degrees ( 200 and 400 mg/kg ) were administered orally. The animate beings were held in place by a suited restrainer with the tail widening out and the tail ( up to 6 centimeter ) was taken dipped in a beaker of H2O maintained at 56 A± 40C. The clip in sec taken to retreat the tail clearly out of H2O was taken as the reaction clip. The first reading ( 0 min ) was taken instantly after the disposal of the trial compound and subsequent reaction clip was recorded at 0, 15, 30, 60 and120 min after the disposal of compounds. A cut off point of 15 sec was observed to forestall the tail harm. The per centum analgetic activity was calculated utilizing the undermentioned expression and the consequences are presented in table 5.3.1.

PAA = [ ( B-A ) /B ] X 100

Where,

B is the reaction clip ( in sec ) before intervention.

A is the reaction clip ( in sec ) after intervention.

PAA is the per centum analgetic activity.

Table 5.3.1 ANALGESIC ACTIVITY OF THE SYNTHESIZED COMPOUNDS

COMP.

Dose

( mg/k )

0 min

15min

30 min

60 min

120 min

MEAN + SEM

MEAN + SEM

%

MEAN + SEM

%

MEAN + SEM

%

MEAN + SEM

%

G1

200

5.25A±0.44

11.08A±0.33*

48.5

12.58A±0.37

62.6

10.10A±0.40*

53.0

9.34i‚±0.24

48.48

G2

200

5.00A±0.50

11.91A±0.20**

44.6

14.16A±0.30**

57.9

10.08A±0.35*

53.1

11.57i‚±0.27

83.94

G3

200

4.00A±0.28

10.66A±0.49

50.4

13.16A±0.60

60.8

10.33A±0.24

51.9

10.35i‚±0.23*

64.54

G4

200

4.83A±0.24

9.00A±0.36*

58.1

14.00A±0.36**

58.3

10.58A±0.27**

50.8

11.77i‚±0.21*

87.12

G5

200

3.66A±o.21

9.83A±0.30*

54.3

12.33A±0.21**

63.3

9.83A±0.30*

54.3

10.12i‚±0.27*

60.89

G6

200

4.16A±0.42

10.75A±0.44*

50.0

13.41A±0.45*

60.1

10.66A±0.33*

50.4

11.74i‚±0.19*

86.64

G7

200

4.75A±0.31

11.25A±0.31

47.7

13.66A±0.71NS

59.4

11.33A±0.33

47.3

9.35i‚±0.25

48.64

G8

200

4.25A±0.35

11.58A±0.23*

46.1

13.33A±0.33*

60.3

11.00A±0.25*

48.8

11.59i‚±0.27

84.26

G9

200

4.75A±0.38

9.16A±0.30

57.4

12.91A±0.15

61.6

10.91A±0.37

49.3

11.21i‚±0.26**

78.21

G10

200

4.83A±0.47

11.16A±0.42*

48.1

13.58A±0.21**

59.6

10.71A±0.45*

50.2

11.83i‚±0.27**

88.07

G11

200

3.58A±0.23

10.00A±0.51*

53.3

12.83A±0.30*

61.8

10.41A±0.35*

53.6

8.82i‚±0.26

40.22

G12

200

4.58A±0.37

9.50A±0.76NS

55.8

13.25A±0.47*

60.6

10.83A±0.30*

49.6

11.23i‚±0.25

78.53

Control

5.33A±0.21

12.16A±0.30**

63.3

14.50A±0.36**

73.2

11.83A±0.30**

56.3

6.29i‚±0.51

Standard

Pentazocin

20

5.25A±0.44

11.08A±0.33*

48.5

12.58A±0.37

62.6

10.10A±0.40*

53.0

7.41i‚±0.32

17.80

RESULTS AND DISCUSSION

Synthesized compounds were evaluated for analgetic activity by tail-immersion method. The activity was studied at 100 and 200 mg/kg b.w. ( p.o ) and consequence was measured at the clip interval of 15, 30, 60 and 120 min.

Most of the synthesized compounds showed important analgetic activity and besides the graded dosage response was observed.

Highest analgetic activity was observed at 60 min for all the compounds ( 100 and 200 mg/kg ) .

When compared with standard drug ( Pentazocin 20 mg/kg, i.p ) the compounds G6, G10, G2 and G4 exhibited comparable analgetic activity at 100 and 200 mg/kg b.w.

Compounds G1, G5 and G3 exhibited moderate analgetic activity.

Among the compounds synthesized compound G11 exhibited lowest analgetic activity.

The order of analgetic activity of the synthesized compounds was shown

G10 & gt ; G4 & gt ; G6 & gt ; G8 & gt ; G2 & gt ; G12 & gt ; G9 & gt ; G3 & gt ; G5 & gt ; G1 & gt ; G7 & gt ; G11

Compounds G10 exhibited a statistically important analgetic activity in the tail submergence method trial in the mice, being nevertheless less active than the equitoxic dosage of pentazocin ( 20 mg/kg ) .

It appears sensible to propose that the presence of o-hydroxy, methoxy substituted on peculiarly 2, 5 place of 1, 3, 4-oxadiazole exhibited better activity.