Title of Invention	SYNTHESIS OF -1-DIPHENYLAMINO-METHYL-3-(4-BROMO-PHENYLIMINO)-5-CHLORO-1,3-DIHYDRO-INDOL-3-ONE AS POTENTIAL NON-STEROIDAL ANTI-INFLAMMATORY AGENT
Abstract	1. The process for the preparation of the compound entitled " 1 -diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one" involving the following steps: step a) reaction between equimolar quantities of 5-chloro-indoline-2,3-dione and 4-bromoaniline dissolved in warm ethanol, refluxed for 30 minutes and standing for approximately 24 hours at room temperature to yield 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step b) reaction between equimolar quantities of diphenylamine in 10 ml of ethanol added to a slurry containing the product of step a and formaldehyde solution in 10 ml of ethanol, stirring for one hour at room temperature and refrigeration for 48 hours to yield l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step c) the product of step b was purified by standard methods.

Title of Invention

SYNTHESIS OF -1-DIPHENYLAMINO-METHYL-3-(4-BROMO-PHENYLIMINO)-5-CHLORO-1,3-DIHYDRO-INDOL-3-ONE AS POTENTIAL NON-STEROIDAL ANTI-INFLAMMATORY AGENT

Abstract

1. The process for the preparation of the compound entitled " 1 -diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one" involving the following steps: step a) reaction between equimolar quantities of 5-chloro-indoline-2,3-dione and 4-bromoaniline dissolved in warm ethanol, refluxed for 30 minutes and standing for approximately 24 hours at room temperature to yield 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step b) reaction between equimolar quantities of diphenylamine in 10 ml of ethanol added to a slurry containing the product of step a and formaldehyde solution in 10 ml of ethanol, stirring for one hour at room temperature and refrigeration for 48 hours to yield l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step c) the product of step b was purified by standard methods.

Full Text	The present invention relating to "Synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one as potential Non-steroidal anti-inflammatory agent" Analgesic Drugs are agents that reduce the sensation of pain. They are broadly classified into opiod analgesics and non-opiod analgesics. Non-opiod analgesics are usually associated with anti-inflammatory and antipyretic properties. They are collectively called as Non-steroidal anti-inflammatory drugs (NSAIDs). They are classified as follows. I. Non-Selective COX Inhibitors 1) Salicylic acid derivatives: Aspirin, Sodium salicylate, Choline magnesium trisalicylate, Salsalate, Diflunisal, Sulfasalazine, Olsalazine. 2) Para-amino phenol derivatives: Acetaminophen 3) Indole & Indene acetic acids: Indomethacin, Sulindac 4) Heteroaryl acetic acids: Tolmetin, Diclofenac, Ketorolac 5) Arylpropionic acids: Ibuprofen, Naproxen, Flurbiprofen, Ketoprofen, Fenoprofen, Oxaprozin 6) Anthranilic acids (fenamates): Mefenamic acid, Meclofenamic acid 7) Enolic acids (oxicams): Piroxicam, Meloxicam 8) Alkanones: Nabumetone 9) Pyrazolone derivatives: Phenylbutazone, Oxyphenbutazone, Antipyrine, Aminopyrine, Dipyrone. n. Selective COX-2 Inhibitors 1) Diary 1 substituted furnanones: Rofecoxib . 2) Diaryl substituted pyrazoles: Celecoxib 3) Indole acetic acid: Etodolac 4) Sulfonanilides: Nimesnildes NSAIDS are used for relieving mild to moderate pain of varied origin, but is not effective for severe pain eg., myocardial infarction or renal or biliary colic. NSAIDs may be combined with opiod analgesic for treatment of cancer pain, where their anti¬inflammatory effects act synergistically with the opiods to enhance analgesia. The anti¬inflammatory properties of NSAIDs are responsible for their are in the treatment of rheumatoid arthritis, rheumatic fever and other inflammatory joint conditions. Certain NSAIDs are used as effective antipyretic agents. At the usual therapeutic dose, many NSAIDS causes gastric upset (intolerance). The gastritis that occurs may be due to irritation of the gastric mucosa by the undissolved tablet or to inhibition of production of protective prostaglandins. Experimental and epidemiological studies overwhelmingly document an increased incident of gastric ulcers and to a lesser extend duodenal ulcers with regular consumption of NSAIDS in chronic condition. Upper gastrointestinal bleeding associated with NSAIDs (aspirin) use is usually related to erosive gastritis. Consumption of higher doses of NSAIDs (salicylates), patients may experience "salicylism" - vomiting, tinnitus, decreased hearing and vertigo which are reversed by reducing the dose. The objective of the invention is to synthesize a novel NSAID with reduced gastric-irritant property. The present invention relates to the synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one. The compound was evaluated for the analgesic, anti-inflammatory and antipyretic activity. The gastric-irritant property of the compound was evaluated. 3-(4-bromo-phenylimino)-5-chloro-1,3-dihydro-indol-2-one was synthesized by reaction of 5-chloro-indoline-2,3-dione and 4-bromoaniline. 1-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-1,3-dihydro-indol-2-one was synthesized by reaction of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one, formaldehyde and diphenylamine. The chemical structure of the synthesized compound was confirmed by means of their Infra-Red, 'H-Nuclear Magnetic Resonance, Mass spectral data and elemental analysis. The synthesized compound was tested for its analgesic, anti-inflammatory and antipyretic activity by standard methods and found to be significantly active. The compound was completely devoid of gastric-irritant property. 1. Synthesis of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one The following methods were employed for the synthesis of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one A suspension of 5-chloro-indoline-2,3-dione (0.01 moles) in 25ml of methanol containing glacial acid (1 drop) was heated on a water bath. 4-Bromoaniline (0.01 moles) was then added to it and heating continued for 2 hours. The reaction mixture was then cooled and the resulting solid filtered and washed with methanol (2x5 ml) to yield the product. 5-Chloro-indoline-2,3-dione (0.01 moles) and the 4-bromoaniline (0.01 moles) were suspending in a mixture of ethanol (15 ml) and dimethylsulphoxide (15ml). The mixture was refluxed for one hour and 30 ml of warm water was added and the mixture was allowed to stand at room temperature overnight. The material was removed by filtration, washed with aqueous ethanol (50% v/v) was dried. 5-Chloro-indoline-2,3-dione (0.01 moles) and the 4-bromoaniline (0.01 moles) were suspending in a mixture of ethanol (15 ml) and dimethylsulphoxide (15ml). The mixture was refluxed for one hour and 30 ml of warm water containing glacial acetic acid (3 drops) was added. The mixture was allowed to stand at room temperature v overnight. The material was removed by filtration, mashed with aqueous ethanol (50% v/v) was dried. 5-Chloro-indoline-2,3-dione and the 4-bromoaniline (0.01 moles) were stirred overnight in dimethylsulphoxide (25ml) containing a few drops of glacial acetic acid. Warm water (30 ml) was added and the mixture was allowed to stand at room temperature overnight. The material was removed by filtration, washed with aqueous ethanol (50% v/v) was dried. Equimolar quantity (0.01 moles) of 5-chloro-indoline-2,3-dione and the 4-bromoaniline. Hydrochloride was dissolved in warm ethanol and heated on a steam bath for 20-40 min. After standing for approximately for 24 hours at room temperature. The products were collected by filtration. Out of the methods employed, the following method was found to produce the desired product with satisfactory yield. Equimolar quantity (0.004 moles) of 5-chloro-indoline-2,3-dione and 4-bromoaniline was dissolved in 10 ml of warm ethanol and refluxed for 30 minutes. After standing for approximately 24 hours at room temperature, the product was separated by filtration, vacuum dried and recrystallized from ethanol. The physico-chemical properties and spectral properties of the compound is as follows., Molecular formula: CuHgBrClN^O Molecular weight: 335.59 Yield: 88 % Melting Point: 234-236 °C Rf value: 0.746 (TLC-Silica gel - Benzene : Chloroform (55:45) - Iodine chamber visualization) Rm value: -0.4679 UV Xrmx (nra): 208, 244, 258, 314, 374. IRCcm"1): 3262(enolic O-H), 1611(CO), 1580(C=N), 1461(C=C), 829, 790, 748(Ar- H); 669(C-Br), 614(C-€1). 'H-NMR (8, ppm): 6.67 - 7.07(s, 1H, NH), 7.13 - 7.73(m, 7H, Ar-H). Mass (m/z): 335 (M+) Elemental Analysis: Found(calculated)., C-50.10(50.06), H-2.40(2.43), N-8.35(8.39). 2. Synthesis of l-diphenyIamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one The following methods were employed for the synthesis of 1-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one To equimolar quantities (0.05 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and diphenylamine in 75 ml of ethanol, 0.05 moles of formaldehyde was added. The reaction mixture was refluxed for 1 hour and cooled to 8° C and maintained for 1 hour. To equimolar quantities (0.01 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and diphenylamine in 25 ml of ethanol, 0.01 moles of formaldehyde was added. The reaction mixture was maintained for 3 hour at room temperature and cooled to 8° C and maintained for 3 hours. Equimolar quantities (0.01 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one, diphenylamine and formaldehyde in 25 ml of ethanol was refluxed for 2 hours. Hot water was added to incipient turbidity and the mixture was cooled to 8°C causing separation of some product. The filtrate was concentrated to about 10ml, hot water was added to faint turbidity and the mixture was cooled overnight. In a 3 necked flask, a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 20ml of water and diphenylamine (0.03 moles) was placed. The mixture was stirred and formaldehyde (0.03 moles) was added slowly. The mixture was refluxed for 2 hours, cooled and the solution was concentrated by distillation on a water bath. To this residue, 25ml of ethanol was added and the solution was concentrated by further distillation. After 12 hours, the product was separated. To 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 15ml of ethanol, formaldehyde (0.03 moles) and dipheny lamine (0.03 moles) was added at 0 °C. The product was recrystallized from absolute alcohol. 3-(4-bromo-phenylimino)'5-chloro-l,3-dihydro-indol-2-one (0.01 moles) in 15ml of ethanol and formaldehyde (0.01 moles) was cooled to 0 °C. Diphenylamine (0.01 moles) was added drop wise with stirring to yield the product. 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.01 moles) in 15ml of ethanol and formaldehyde (0.01 moles) was cooled to 0 °C. Diphenylamine (0.01 moles) was added drop wise. The reaction mixture was cooled to 0 °C and maintained for 12 hours. To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 20ml of water, diphenylamine (0.03 moles) was added and formaldehyde (0.03 moles) was introduced drop wise. The reaction mixture was heated to 85 °C for 14 min and to 90 °C for 30 min with stirring. The contents were cooled and extracted from the cold solution with chloroform. To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 50% ethanol (5 ml), formaldehyde (0.03 moles) and diphenylamine was added drop wise with cooling and shaking. The contents was wanned on a steam bath for 15 min and cooled to room temperature. To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 50% ethanol (5 ml), formaldehyde (0.03 moles) and diphenylamine was added drop wise with cooling and shaking. The contents were cooled to 8 °C for 1 hour. A solution of diphenylamine (0.25 moles) in 10ml of ethanol was added to a slurry containing 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.25 moles) and formaldehyde (0.25 moles) in 10ml of ethanol. The reaction mixture was stirred for 30 min and refrigerated (8 °C) for 12 hours. To 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.01 moles), diphenylamine (0.01 moles) and formaldehyde (0.01 moles) in 10ml of ethanol and 10ml of dimethylsulphoxide. The contents were heated until a clear solution was obtained. 10 ml of hot water was added and allowed to stand for 12 hours. To a clear boiling solution 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in dimethylformamide-methanol (10ml), formaldehyde (0.03 moles) and diphenylamine (0.03 moles) was added. The contents were heated with stirring for 10 min and allowed to stand for 18 hours. Out of the methods employed, the following method was found to produce the desired product with satisfactory yield., Equimolar quantity (0.004 moles) of diphenylamine in 10 ml of ethanol was added to a slurry containing 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and formaldehyde solution in 10 ml of ethanol. The reaction mixture was stirred for one hour at room temperature and refrigerated for 48 hours. The product was separated by suction filtration, vacuurn dried and recrystallized from ethanol. The physico-chemical properties and spectral properties of the compound is as follows., Pharmacological activity The synthesized compound was evaluated for analgesic, antiinflammatory and antipyretic activity. The experimental dose was selected between the minimum effective dose and minimum non-lethal dose. Student-t test was performed for all the activities to ascertain the significance of the exhibited activities. The test compound and the standard drugs was administered in the form of a suspension (1% Carboxy methyl cellulose as vehicle) in the same route of administration. The animals was maintained in colony cages at 25 ± 2 °C, relative humidity of 45-55%, maintained under 12 hour light and dark cycle and was fed with standard animal feed. All the animals was acclimatized for a week before use. 1. Determination of LD50 Acute toxicity test was performed using wistar albino mice (25-30g, intraperitoneal route of administration) for the synthesized compound to ascertain the LD50 values by Karber's arithmetical method and was found to be 878 mg/kg. 2. Analgesic activity The analgesic activity was determined by acetic acid induced writhing method using wistar albino mice (25-30g) of either sex selected by random sampling technique. Paracetamol at a dose level of 100 mg/kg was administered as standard drug for comparison. The test compound at 3 dose levels (25, 50 and lOOmg/kg) was administered intraperitoneally 15 min prior to administration of the writhing agent (0.6% v/v aqueous acetic acid - lml/lOOg). The writhings produced in the animal was observed for 30 minutes and percentage protection was calculated for analgesic activity. The analgesic activity data are presented in table 1. 3. Anti-inflammatory activity The Anti-inflammatory activity was determined by formalin induced pedal paw edema method in wistar albino rat (150-200g) of either sex by using Plethysmograph. Diclofenac sodium (50mg/kg) was administered as standard drug. The test compound was administered at 3 dose levels (100, 200 and 400 mg) intraperitoneally 30 minutes prior to administration of formalin (0.1 ml of 1% w/v) in the plantar region of the paw. The paw volume was measured at 15, 30, 60 and 120 min after formalin administration. The antiinflammatory activity data are presented in table 2. 4. Antipyretic activity The Antipyretic activity was determined by experimentally induced pyrexia in white New Zealand rabbits (0.95-1.1 kg) of either sex. Paracetamol (lOOmg/kg) was administered as standard drug for comparison. The compound at 3 dose levels (50, 100 and 200 mg/kg) was administered subcutaneously prior to administration of 12% w/v of suspended Brewer's yeast (1 ml/lOOg) subcutaneously. After administration of yeast, the rectal temperature of the rabbits was recorded by using telethermometer at 30, 60, 90, 120, 150 & 180 minutes intervals. The antipyretic activity data is presented in table 3. 5. Gastric-irritant property The gastric-irritant property of the compound was tested in pylorus-ligated rats. The compound was found to completely devoid of gastric irritant property. We Claim: 1. The process for the preparation of the compound entitled " 1 -diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one" involving the following steps: step a) reaction between equimolar quantities of 5-chloro-indoline-2,3-dione and 4-bromoaniline dissolved in warm ethanol, refluxed for 30 minutes and standing for approximately 24 hours at room temperature to yield 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step b) reaction between equimolar quantities of diphenylamine in 10 ml of ethanol added to a slurry containing the product of step a and formaldehyde solution in 10 ml of ethanol, stirring for one hour at room temperature and refrigeration for 48 hours to yield l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step c) the product of step b was purified by standard methods. 2. A process "Synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one as potential Non-steroidal anti-inflammatory agent" is herein substantially defined and exemplified.

Full Text

The present invention relating to "Synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one as potential Non-steroidal anti-inflammatory agent"
Analgesic Drugs are agents that reduce the sensation of pain. They are broadly classified into opiod analgesics and non-opiod analgesics. Non-opiod analgesics are usually associated with anti-inflammatory and antipyretic properties. They are collectively called as Non-steroidal anti-inflammatory drugs (NSAIDs). They are classified as follows.
I. Non-Selective COX Inhibitors
1) Salicylic acid derivatives: Aspirin, Sodium salicylate, Choline magnesium
trisalicylate, Salsalate, Diflunisal, Sulfasalazine, Olsalazine.
2) Para-amino phenol derivatives: Acetaminophen
3) Indole & Indene acetic acids: Indomethacin, Sulindac
4) Heteroaryl acetic acids: Tolmetin, Diclofenac, Ketorolac
5) Arylpropionic acids: Ibuprofen, Naproxen, Flurbiprofen, Ketoprofen,
Fenoprofen, Oxaprozin
6) Anthranilic acids (fenamates): Mefenamic acid, Meclofenamic acid
7) Enolic acids (oxicams): Piroxicam, Meloxicam
8) Alkanones: Nabumetone
9) Pyrazolone derivatives: Phenylbutazone, Oxyphenbutazone, Antipyrine,
Aminopyrine, Dipyrone.
n. Selective COX-2 Inhibitors
1) Diary 1 substituted furnanones: Rofecoxib . 2) Diaryl substituted pyrazoles: Celecoxib
3) Indole acetic acid: Etodolac
4) Sulfonanilides: Nimesnildes
NSAIDS are used for relieving mild to moderate pain of varied origin, but is not effective for severe pain eg., myocardial infarction or renal or biliary colic. NSAIDs may be combined with opiod analgesic for treatment of cancer pain, where their anti¬inflammatory effects act synergistically with the opiods to enhance analgesia. The anti¬inflammatory properties of NSAIDs are responsible for their are in the treatment of rheumatoid arthritis, rheumatic fever and other inflammatory joint conditions. Certain NSAIDs are used as effective antipyretic agents.
At the usual therapeutic dose, many NSAIDS causes gastric upset (intolerance). The gastritis that occurs may be due to irritation of the gastric mucosa by the undissolved tablet or to inhibition of production of protective prostaglandins. Experimental and epidemiological studies overwhelmingly document an increased incident of gastric

ulcers and to a lesser extend duodenal ulcers with regular consumption of NSAIDS in chronic condition. Upper gastrointestinal bleeding associated with NSAIDs (aspirin) use is usually related to erosive gastritis. Consumption of higher doses of NSAIDs (salicylates), patients may experience "salicylism" - vomiting, tinnitus, decreased hearing and vertigo which are reversed by reducing the dose.
The objective of the invention is to synthesize a novel NSAID with reduced gastric-irritant property.
The present invention relates to the synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one. The compound was evaluated for the analgesic, anti-inflammatory and antipyretic activity. The gastric-irritant property of the compound was evaluated.
3-(4-bromo-phenylimino)-5-chloro-1,3-dihydro-indol-2-one was synthesized by reaction of 5-chloro-indoline-2,3-dione and 4-bromoaniline. 1-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-1,3-dihydro-indol-2-one was synthesized by reaction of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one, formaldehyde and diphenylamine. The chemical structure of the synthesized compound was confirmed by means of their Infra-Red, 'H-Nuclear Magnetic Resonance, Mass spectral data and elemental analysis. The synthesized compound was tested for its analgesic, anti-inflammatory and antipyretic activity by standard methods and found to be significantly active. The compound was completely devoid of gastric-irritant property.
1. Synthesis of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one
The following methods were employed for the synthesis of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one
A suspension of 5-chloro-indoline-2,3-dione (0.01 moles) in 25ml of methanol containing glacial acid (1 drop) was heated on a water bath. 4-Bromoaniline (0.01 moles) was then added to it and heating continued for 2 hours. The reaction mixture was then cooled and the resulting solid filtered and washed with methanol (2x5 ml) to yield the product.
5-Chloro-indoline-2,3-dione (0.01 moles) and the 4-bromoaniline (0.01 moles) were suspending in a mixture of ethanol (15 ml) and dimethylsulphoxide (15ml). The mixture was refluxed for one hour and 30 ml of warm water was added and the mixture was allowed to stand at room temperature overnight. The material was removed by filtration, washed with aqueous ethanol (50% v/v) was dried.
5-Chloro-indoline-2,3-dione (0.01 moles) and the 4-bromoaniline (0.01 moles) were suspending in a mixture of ethanol (15 ml) and dimethylsulphoxide (15ml). The mixture was refluxed for one hour and 30 ml of warm water containing glacial acetic acid (3 drops) was added. The mixture was allowed to stand at room temperature
v

overnight. The material was removed by filtration, mashed with aqueous ethanol (50% v/v) was dried.
5-Chloro-indoline-2,3-dione and the 4-bromoaniline (0.01 moles) were stirred overnight in dimethylsulphoxide (25ml) containing a few drops of glacial acetic acid. Warm water (30 ml) was added and the mixture was allowed to stand at room temperature overnight. The material was removed by filtration, washed with aqueous ethanol (50% v/v) was dried.
Equimolar quantity (0.01 moles) of 5-chloro-indoline-2,3-dione and the 4-bromoaniline. Hydrochloride was dissolved in warm ethanol and heated on a steam bath for 20-40 min. After standing for approximately for 24 hours at room temperature. The products were collected by filtration.
Out of the methods employed, the following method was found to produce the desired product with satisfactory yield.
Equimolar quantity (0.004 moles) of 5-chloro-indoline-2,3-dione and 4-bromoaniline was dissolved in 10 ml of warm ethanol and refluxed for 30 minutes. After standing for approximately 24 hours at room temperature, the product was separated by filtration, vacuum dried and recrystallized from ethanol. The physico-chemical properties and spectral properties of the compound is as follows.,
Molecular formula: CuHgBrClN^O
Molecular weight: 335.59
Yield: 88 %
Melting Point: 234-236 °C
Rf value: 0.746 (TLC-Silica gel - Benzene : Chloroform (55:45) - Iodine chamber
visualization)
Rm value: -0.4679
UV Xrmx (nra): 208, 244, 258, 314, 374.
IRCcm"1): 3262(enolic O-H), 1611(CO), 1580(C=N), 1461(C=C), 829, 790, 748(Ar-
H); 669(C-Br), 614(C-€1).
'H-NMR (8, ppm): 6.67 - 7.07(s, 1H, NH), 7.13 - 7.73(m, 7H, Ar-H).
Mass (m/z): 335 (M+)
Elemental Analysis: Found(calculated)., C-50.10(50.06), H-2.40(2.43), N-8.35(8.39).

2. Synthesis of l-diphenyIamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one
The following methods were employed for the synthesis of 1-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one
To equimolar quantities (0.05 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and diphenylamine in 75 ml of ethanol, 0.05 moles of formaldehyde was added. The reaction mixture was refluxed for 1 hour and cooled to 8° C and maintained for 1 hour.
To equimolar quantities (0.01 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and diphenylamine in 25 ml of ethanol, 0.01 moles of formaldehyde was added. The reaction mixture was maintained for 3 hour at room temperature and cooled to 8° C and maintained for 3 hours.
Equimolar quantities (0.01 moles) of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one, diphenylamine and formaldehyde in 25 ml of ethanol was refluxed for 2 hours. Hot water was added to incipient turbidity and the mixture was cooled to 8°C causing separation of some product. The filtrate was concentrated to about 10ml, hot water was added to faint turbidity and the mixture was cooled overnight.
In a 3 necked flask, a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 20ml of water and diphenylamine (0.03 moles) was placed. The mixture was stirred and formaldehyde (0.03 moles) was added slowly. The mixture

was refluxed for 2 hours, cooled and the solution was concentrated by distillation on a water bath. To this residue, 25ml of ethanol was added and the solution was concentrated by further distillation. After 12 hours, the product was separated.
To 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 15ml of ethanol, formaldehyde (0.03 moles) and dipheny lamine (0.03 moles) was added at 0 °C. The product was recrystallized from absolute alcohol.
3-(4-bromo-phenylimino)'5-chloro-l,3-dihydro-indol-2-one (0.01 moles) in 15ml of ethanol and formaldehyde (0.01 moles) was cooled to 0 °C. Diphenylamine (0.01 moles) was added drop wise with stirring to yield the product.
3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.01 moles) in 15ml of ethanol and formaldehyde (0.01 moles) was cooled to 0 °C. Diphenylamine (0.01 moles) was added drop wise. The reaction mixture was cooled to 0 °C and maintained for 12 hours.
To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 20ml of water, diphenylamine (0.03 moles) was added and formaldehyde (0.03 moles) was introduced drop wise. The reaction mixture was heated to 85 °C for 14 min and to 90 °C for 30 min with stirring. The contents were cooled and extracted from the cold solution with chloroform.
To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 50% ethanol (5 ml), formaldehyde (0.03 moles) and diphenylamine was added drop wise with cooling and shaking. The contents was wanned on a steam bath for 15 min and cooled to room temperature.
To a slurry of 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in 50% ethanol (5 ml), formaldehyde (0.03 moles) and diphenylamine was added drop wise with cooling and shaking. The contents were cooled to 8 °C for 1 hour.
A solution of diphenylamine (0.25 moles) in 10ml of ethanol was added to a slurry containing 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.25 moles) and formaldehyde (0.25 moles) in 10ml of ethanol. The reaction mixture was stirred for 30 min and refrigerated (8 °C) for 12 hours.
To 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.01 moles), diphenylamine (0.01 moles) and formaldehyde (0.01 moles) in 10ml of ethanol and 10ml of dimethylsulphoxide. The contents were heated until a clear solution was obtained. 10 ml of hot water was added and allowed to stand for 12 hours.
To a clear boiling solution 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one (0.03 moles) in dimethylformamide-methanol (10ml), formaldehyde (0.03 moles) and diphenylamine (0.03 moles) was added. The contents were heated with stirring for 10 min and allowed to stand for 18 hours.

Out of the methods employed, the following method was found to produce the desired product with satisfactory yield.,
Equimolar quantity (0.004 moles) of diphenylamine in 10 ml of ethanol was added to a slurry containing 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one and formaldehyde solution in 10 ml of ethanol. The reaction mixture was stirred for one hour at room temperature and refrigerated for 48 hours. The product was separated by suction filtration, vacuurn dried and recrystallized from ethanol. The physico-chemical properties and spectral properties of the compound is as follows.,

Pharmacological activity
The synthesized compound was evaluated for analgesic, antiinflammatory and antipyretic activity. The experimental dose was selected between the minimum effective dose and minimum non-lethal dose. Student-t test was performed for all the activities to ascertain the significance of the exhibited activities. The test compound and the standard drugs was administered in the form of a suspension (1% Carboxy methyl cellulose as vehicle) in the same route of administration. The animals was maintained in colony cages at 25 ± 2 °C, relative humidity of 45-55%, maintained under 12 hour light and dark cycle and was fed with standard animal feed. All the animals was acclimatized for a week before use.
1. Determination of LD50
Acute toxicity test was performed using wistar albino mice (25-30g, intraperitoneal route of administration) for the synthesized compound to ascertain the LD50 values by Karber's arithmetical method and was found to be 878 mg/kg.
2. Analgesic activity
The analgesic activity was determined by acetic acid induced writhing method using wistar albino mice (25-30g) of either sex selected by random sampling technique. Paracetamol at a dose level of 100 mg/kg was administered as standard drug for comparison. The test compound at 3 dose levels (25, 50 and lOOmg/kg) was administered intraperitoneally 15 min prior to administration of the writhing agent (0.6% v/v aqueous acetic acid - lml/lOOg). The writhings produced in the animal was observed for 30 minutes and percentage protection was calculated for analgesic activity. The analgesic activity data are presented in table 1.
3. Anti-inflammatory activity
The Anti-inflammatory activity was determined by formalin induced pedal paw edema method in wistar albino rat (150-200g) of either sex by using Plethysmograph. Diclofenac sodium (50mg/kg) was administered as standard drug. The test compound was administered at 3 dose levels (100, 200 and 400 mg) intraperitoneally 30 minutes prior to administration of formalin (0.1 ml of 1% w/v) in the plantar region of the paw. The paw volume was measured at 15, 30, 60 and 120 min after formalin administration. The antiinflammatory activity data are presented in table 2.
4. Antipyretic activity
The Antipyretic activity was determined by experimentally induced pyrexia in white New Zealand rabbits (0.95-1.1 kg) of either sex. Paracetamol (lOOmg/kg) was administered as standard drug for comparison. The compound at 3 dose levels (50, 100 and 200 mg/kg) was administered subcutaneously prior to administration of 12% w/v of suspended Brewer's yeast (1 ml/lOOg) subcutaneously. After administration of yeast, the rectal temperature of the rabbits was recorded by using telethermometer at 30, 60, 90, 120, 150 & 180 minutes intervals. The antipyretic activity data is presented in table 3.

5. Gastric-irritant property
The gastric-irritant property of the compound was tested in pylorus-ligated rats. The compound was found to completely devoid of gastric irritant property.

We Claim:
1. The process for the preparation of the compound entitled " 1 -diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one" involving the following steps: step a) reaction between equimolar quantities of 5-chloro-indoline-2,3-dione and 4-bromoaniline dissolved in warm ethanol, refluxed for 30 minutes and standing for approximately 24 hours at room temperature to yield 3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step b) reaction between equimolar quantities of diphenylamine in 10 ml of ethanol added to a slurry containing the product of step a and formaldehyde solution in 10 ml of ethanol, stirring for one hour at room temperature and refrigeration for 48 hours to yield l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one; step c) the product of step b was purified by standard methods.
2. A process "Synthesis of l-diphenylamino-methyl-3-(4-bromo-phenylimino)-5-chloro-l,3-dihydro-indol-2-one as potential Non-steroidal anti-inflammatory agent" is herein substantially defined and exemplified.

Documents:

0728-mas-2001 claims.pdf

0728-mas-2001 correspondence-others.pdf

0728-mas-2001 correspondence-po.pdf

0728-mas-2001 description (complete).pdf

0728-mas-2001 form-1.pdf

« Previous Patent

Next Patent »

Patent Number

193623

Indian Patent Application Number

728/MAS/2001

PG Journal Number

31/2012

Publication Date

03-Aug-2012

Grant Date

06-Dec-2005

Date of Filing

05-Sep-2001

Name of Patentee

SESHAIAH KRISHNAN SRIDHAR

Applicant Address

NO.19  FIRST STREET  BALAJI NAGAR  ROYAPETTAH  CHENNAI-600 014

Inventors:

#	Inventor's Name	Inventor's Address
1	SESHAIAH KRISHNAN SRIDHAR	NO.19, FIRST STREET, BALAJI NAGAR, ROYAPETTAH, CHENNAI-600 014
2	ATMAKURU RAMESH	NO.184, RANGARAJAPURAM MAIN ROAD (NEAR RANGARAJAPURAM RAILWAY GATE), KODAMBAKKAM, CHENNAI-600 024

PCT International Classification Number

C07D209/04

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA