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Title:
ARYL AZO PYRAZOLE DERIVATIVES AS ANTIMICROBIAL AGENT
Document Type and Number:
WIPO Patent Application WO/2023/209678
Kind Code:
A1
Abstract:
The present invention is a compound of Formula (I), or a pharmaceutically acceptable salt thereof. The present invention also relates to the derivatives of the said compound of Formula (I). The present invention relates to method of preparing the compounds and the use of those compounds in preventing microbial infection and inflammation.

Inventors:
PATEL ARPAN B (US)
Application Number:
PCT/IB2023/054471
Publication Date:
November 02, 2023
Filing Date:
April 28, 2023
Export Citation:
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Assignee:
ZENAL PHARMACHEM LLC (US)
International Classes:
C07D231/46; A61K31/4152; A61P29/00; A61P31/04
Foreign References:
US20120189670A12012-07-26
US20130184317A12013-07-18
US20100056581A12010-03-04
Other References:
DATABASE PUBCHEM SUBSTANCE ANONYMOUS : "4-((1-benzoyl-3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-4-yl)diazenyl)-N-(pyrimidin-2-yl)benzenesulfonamide", XP093106791, retrieved from PUBCHEM
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Claims:
CLAIMS We claim, 1. An aryl azo pyrazole derivatives, comprising the compound of structural formula I or a pharmaceutically acceptable salt thereof, in which R1 is an alkoxy or a halo group. 2. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the alkoxy group is selected from methoxy, ethoxy, isopropoxy, tert-butoxy and phenoxy, more preferably a methoxy. 3. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the halo group is selected from flouro, chloro, bromo and iodine, more preferably a flouro. 4. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the derivative is (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl-5-oxo-1,5-dihydro- 4H-pyrazol-4-ylidene)hydrazinyl)-N-(pyrimidin-2- yl)benzenesulfonamide: 5. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the another derivative is (E)-4-(2-(1-(4-fluorobenzoyl)-3-methyl-5-oxo-1, 5- dihydro- 4H-pyrazol-4-ylidene)hydrazinyl)-N-(pyrimidin-2- yl)benzenesulfonamide: 6. A process for preparation of compound of Formula-I as claimed in claim 1, comprising the following steps: a) Dissolving 100g of 0.4mol of sulfadiazine in 700ml solution of hydrochloric acid and cool the mixture to 0-5° C. b) Dissolving 41g of sodium nitrite in 200 ml of water and dropwise adding the solution into the mixture of step (a). c) Stirring the mixture of step (b) at 0-5° C for 30 minutes. d) Dissolving 238 g of sodium acetate in 600ml of water and stir to form a mixture. e) Dissolving 54.6g of 0.42mol ethyl acetoacetate to the solution of step (d) and maintaining the mixture at 0-5° C. f) Dropwise adding the mixture of step (c) into the mixture of step (e) while keeping the temperature 0-5 °C within 30 minutes. g) Stirring the reaction mixture of step (f) for 30 minutes. h) Filtering the reaction mixture of step (g) and washing it with 300ml of cold water. i) Drying the compound under vacuum at 45-50°C to get intermediate 3. j) Dissolving 84.8g of the intermediate 3 of step (i) and 36.0g of benzohydrazide into 840ml of acetic acid. k) Stirring the mixture of step (j) at 25-300C for 30 minutes and then heat it at 95-1000C under stirring for 40 hours. l) Cooling the mixture of step (k) to 400C and keep stirring at the same temperature for 1 hour. m) Filter the precipitated compound of step (l) and washing it with 200 ml of cold water. n) Drying the compound of step (m) under vacuum at 45-500C to get 30.9g of the crude compound. o) Suspending 30.9g of the crude compound of step (n) in 600ml of methanol and reflux the mixture under stirring for 2-3 hours. p) Cool the suspension of step (o) to 25-300C and is stirred for 1 hour. q) Filtering the compound of step (p) and washing it with 200ml of methanol and drying the compound under vacuum at 45-500C to get the pure compound with formula (I). 7. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the said compound is used to prepare pharmaceutical composition comprising a pharmaceutically active compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. 8. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the pharmaceutical composition is used as an anti-microbial agent for treating microbial infection. 9. The aryl azo pyrazole derivatives as claimed in claim 1, wherein the said derivatives having LD504000mg/kg.
Description:
ARYL AZO PYRAZOLE DERIVATIVES AS ANTIMICROBIAL AGENT FIELD OF THE INVENTION The invention relates to compound of formula (I), or a pharmaceutically acceptable salt thereof. More particularly, the present invention relate to method of synthesizing the compounds and the use of those compounds in treating, ameliorating, or preventing a microbial infection and inflammation. BACKGROUND OF THE INVENTION An antimicrobial agent is an agent that kills microorganisms or stops their growth. Antimicrobial medicines can be grouped according to the microorganisms they act primarily against. For example, antibiotics are used against bacteria, and antifungals are used against fungi. They can also be classified according to their function. Agents that kill microbes are microbicides, while those that merely inhibit their growth are called bacteriostatic agents. The discovery of the first selective antimicrobial agent approximately four decades ago was a major milestone in the history of medicine and human health. The subsequent development of antimicrobial therapy largely centered on the search for drugs with effectiveness against microbial species that were not susceptible to drugs then in use. The various bacterial diseases such as tuberculosis and fungal diseases are refractory to known antibiotic agents. Development of novel antimicrobial compounds is a continuing urgent public health need. Widespread use of antimicrobial agents in recent decades has led to proliferation of pathogens having multiple drug resistance, often encoded by transmissible plasmids, and therefore capable of spreading rapidly between species. Many previously useful antimicrobial agents are no longer effective against infectious organisms isolated from human and animal subjects. The specter of epidemic forms of bacterial diseases such as tuberculosis and fungal diseases, which are refractory to known antibiotic agents, may be realized in the near future. The discovery and development of new antimicrobial agents is a major focus of many pharmaceutical companies. However, in more recent years this area of research and drug development is neglected resulting in very few new antimicrobials entering the market. This lack of new antimicrobial is particularly disturbing, especially at a time when bacterial resistance to current therapies is increasing both in the hospital and community settings. Thus, there is a need to develop new antimicrobial compounds. The present invention arose due to the inventor's interest in pyrazole heterocyclic moiety and developed novel Aryl azo Pyrazole derivatives as Antimicrobial Agent. OBJECTIVES OF THE INVENTION The main objective of the present invention is to provide aryl azo pyrazole derivatives, compounds having the structural formula (I). Another objective of the present invention is to provide aryl azo pyrazole derivatives (I) which can be used as antimicrobial and anti-inflammatory agent. Another objective of the present invention is to provide a process of synthesizing the aryl azo pyrazole derivatives (I) SUMMARY OF THE INVENTION The main aspect of the present invention, is to provide aryl azo pyrazole derivatives, compounds represented by formula (I) or a pharmaceutical acceptable salt thereof; Wherein R 1 is methoxy or fluoro. Another aspect of the present invention is to provide a derivative, wherein the derivative is (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H- pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide represented by formula (I-1) According to another aspect of the present invention, the derivative is (E)-4-(2-(1- (4-fluorobenzoyl)-3-methyl-5-oxo-1, 5- dihydro-4H-pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide represented by the formula (I- 2) Another aspect of the present invention is to provide the process for preparing the compounds and derivatives having formula (I). Another aspect of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. Another aspect of the present invention is to provide the process of synthesizing the compound of formula (I) and the use of it in treating, ameliorating, or preventing a microbial infection or inflammation. BREIF DESCRIPTION OF DRAWING Figure 1 illustrates FT-IR results of Formula (I-1) Figure 2 illustrates FT-IR results of Formula (I-2) Figure 3 illustrates Mass spec results of Formula (I-1) Figure 4 illustrates Mass spec results of Formula (I-2) Figure 5 illustrates NMR results of Formula (I-1) Figure 6 illustrates NMR results of Formula (I-2) DETAILED DESCRIPTION OF THE INVENTION The present invention is about aryl azo pyrazole derivatives as antimicrobial agent, as described herein. The phrase “pharmaceutically acceptable salt(s)”, as used herein, unless otherwise indicated, includes salts of acidic or basic groups which may be present in the compounds of the present invention. The compounds of the present invention that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that may be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as the hydrochloride, hydrobromide, hydro iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, and ascorbate. Succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, Saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate i.e., 1,1'- methylene-bis-(2- hydroxy-3-naphthoate) salts. The compounds of the present invention that include a basic moiety, such as an amino group, may form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above. The invention also relates to base addition salts of the compounds of the invention. The chemical bases that may be used as reagents to prepare pharmaceutically acceptable base salts of those compounds of the compounds of the invention that are acidic in nature are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to those derived from such pharmacologically acceptable cations such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts such as N-methylglucamine- (meglu mine), and the lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines. Suitable base salts are formed from bases which form non-toxic salts. Non-limiting examples of suitable base salts include the aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, Sodium, tromethamine and Zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulphate and hemicalcium salts. The phrase “Antimicrobial Agent” is agent that kills microorganisms or stops their growth. Microorganisms include bacteria, fungi, virus and parasite. The main embodiment of the present invention relates to a compound represented by formula (I), or a pharmaceutically acceptable salt thereof: Wherein, R 1 is an alkoxy or a halo group. According to another embodiment of the present invention the alkoxy group is selected from methoxy, ethoxy, isopropoxy, tert-butoxy and phenoxy, more preferably a methoxy. According to another embodiment of the present invention the halo group is selected from flouro, chloro, bromo and iodine, more preferably a flouro. In another embodiment of the present invention, the aryl azo pyrazole derivative is (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H-pyrazol-4- ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide, having the formula In another embodiment of the present invention, the aryl azo pyrazole derivative is (E)-4-(2-(1-(4-fluorobenzoyl)-3-methyl-5-oxo-1, 5- dihydro-4H-pyrazol-4- ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide

In another embodiment, the present invention provides a method for the preparation of compound with formula (I), comprising the following steps: a) Dissolving 100g of 0.4mol of sulfadiazine in 700ml solution of hydrochloric acid and cool the mixture to 0-5° C. b) Dissolving 41g of sodium nitrite in 200 ml of water and dropwise adding the solution into the mixture of step (a). c) Stirring the mixture of step (b) at 0-5° C for 30 minutes. d) Dissolving 238 g of sodium acetate in 600ml of water and stir to form a mixture. e) Dissolving 54.6g of 0.42mol ethyl acetoacetate to the solution of step (d) and maintaining the mixture at 0-5° C. f) Dropwise adding the mixture of step (c) into the mixture of step (e) while keeping the temperature 0-5 ° C within 30 minutes. g) Stirring the reaction mixture of step (f) for 30 minutes. h) Filtering the reaction mixture of step (g) and washing it with 300ml of cold water. i) Drying the compound under vacuum at 45-50°C to get the final compound with formula (I). In another embodiment, the present invention provides a compound wherein the said compound is used to prepare a pharmaceutical composition comprising pharmaceutically active compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. Pharmaceutical compositions may be formulated in a conventional manner using one or more physiologically accept able carriers including excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any of the well-known techniques, carriers, and excipients may be used as Suitable and as understood in the art. The pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, and suspension, for parenteral injection as a sterile solution, suspension, or emulsion, for topical administration as an ointment or cream or for rectal administration as a Suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc. Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99% by weight, more preferably from 0.1 to 70% by weight of the active ingredient, and, from 1 to 99.95% by weight, more preferably from 30 to 99.9 weight % of a pharmaceutically acceptable carrier, all percentages being based on the total composition. The pharmaceutical composition may additionally contain various other ingredients known in the art, for example, a lubricant, stabilizing agent, buffering agent, emulsifying agent, viscosity-regulating agent, surfactant, preservative, flavouring or colorant. In another embodiment, the present invention relates to methods of synthesizing the compound of formula (I-1) and the compound of formula (I-2) and the use of it in treating, ameliorating, or preventing a microbial infection. In another embodiment, the present invention relates to compounds of formula (I- 1) and formula (I-2) for treating, ameliorating, or preventing microbial inflammation. In another embodiment, the present invention relates to the compounds of formula (I-1) and formula (I-2) are showing antipyretic activity. Compounds described herein may be in various forms, including but not limited to, amorphous forms, milled forms and nano-particulate forms. In addition, compounds described herein include crystalline forms, also known as polymorphs. Polymorphs include the different crystal packing arrangements of the same elemental composition of a compound. Polymorphs usually have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability, and solubility. Various factors such as the recrystallization Solvent, rate of crystallization, and storage temperature may cause a single crystal form to dominate. EXAMPLES The following working examples represent preferred embodiments of the present invention. All temperatures are expressed in degrees Centigrade unless otherwise indicated. Example 1: Method of synthesis of compound of formula (I):

Step 1: Preparation of ethyl (E)-3-oxo-2-(2-(4-(N-(pyrimidin-2-yl) sulfamoyl) phenyl) hydrazono) butanoate (Intermediate 3): a) Dissolving 100g of 0.4mol of 4-amino-N-(pyrimidin-2-yl) benzenesulfonamide in 700ml solution of hydrochloric acid and cool the mixture to 0-5° C. b) Dissolving 41g of sodium nitrite in 200 ml of water and dropwise adding the solution into the mixture of step (a). c) Stirring the mixture of step (b) at 0-5° C for 30 minutes. d) Dissolving 238 g of sodium acetate in 600ml of water and stir to form a mixture. e) Dissolving 54.6g of 0.42mol ethyl acetoacetate to the solution of step (d) and maintaining the mixture at 0-5° C. f) Dropwise adding the mixture of step (c) into the mixture of step (e) while keeping the temperature 0-5° C within 30minutes. g) Stirring the reaction mixture of step (f) for 30 minutes. h) Filtering the reaction mixture of step (g) and washing it with 300ml of cold water. i) Drying the compound under vacuum at 45-50°C to get Intermediate (3). Step 2: Preparation of compound with Formula (I): a) Dissolving 84.8g of Intermediate 3 and 36.0g of benzohydrazide into 840ml of acetic acid at room temperature. b) Stirring the reaction mixture of step (a) at 25-30 0 C for 30 minutes and then heat it at 95-100 0 C under stirring for 40 hours. c) Cooling the mixture of step (b) to 35-40 0 C and keep stirring at the same temperature for 1 hour. d) Filter the precipitated compound of step (c) and washing it with 200 ml of cold water. e) Drying the compound of step (d) under vacuum at 45-50 0 C to get 30.9g of the crude compound. f) Suspending 30.9g of the crude compound of step (e) in 600ml of methanol and reflux the mixture under stirring for 2-3 hours. g) Cool the suspension of step (f) to 25-30 0 C and is stirred for 1 hour. h) Filtering the compound of step (g) and washing it with 200ml of methanol and drying the compound under vacuum at 45-50 0 C to get the pure compound with formula (I). Example 2: Method of synthesis of derivative of formula (I-1) (E)-4-(2-(1-(4- methoxybenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H-pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide: Step 1: Preparation of (E)-ethyl 3-oxo-2-(2-(4-(N-(pyrimidin 2yl) sulfamoyl) phenyl) hydrazono) butanoate (Intermediate-3): a) Dissolving 100g of 0.4mol of 4-amino-N-(pyrimidin-2-yl) benzenesulfonamide in 700ml solution of hydrochloric acid and cool the mixture to 0-5° C. b) Dissolving 41g of sodium nitrite in 200 ml of water and dropwise adding the solution into the mixture of step (a). c) Stirring the mixture of step (b) at 0-5° C for 30 minutes. d) Dissolving 289g of sodium acetate in 600ml of water and stir to form a mixture. e) Dissolving 54.6g of 0.42mol ethyl acetoacetate to the solution of step (d) and maintaining the mixture at 0-5° C. f) Dropwise adding the mixture of step (c) into the mixture of step (e) while keeping the temperature 0-5° C within 30minutes. g) Stirring the reaction mixture of step (f) for 30 minutes. h) Filtering the reaction mixture of step (g) and washing it with 300ml of cold water. i) Drying the compound under vacuum at 45-50°C to get Intermediate (3). Step 2: Preparation of (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl-5-oxo-1, 5- dihydro-4H-pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide [Formula (I-1)]: a) Dissolving 84.8g of Intermediate 3 and 36.0g of 4- methoxybenzohydrazide into 840ml of acetic acid at room temperature. b) Stirring the reaction mixture of step (a) at 25-30 0 C for 30 minutes and then heat it at 95-100 0 C under stirring for 40 hours. c) Cooling the mixture of step (b) to 35-40 0 C and keep stirring at the same temperature for 1 hour. d) Filter the precipitated compound of step (c) and washing it with 200 ml of cold water. e) Drying the compound of step (d) under vacuum at 45-50 0 C to get 30.9g of the crude compound. f) Suspending 30.9g of the crude compound of step (e) in 600ml of methanol and reflux the mixture under stirring for 2-3 hours. g) Cool the suspension of step (f) to 25-30 0 C and is stirred for 1 hour. h) Filtering the compound of step (g) and washing it with 200ml of methanol and drying the compound under vacuum at 45-50 0 C to get the pure compound with formula (I-1) (E)-4-(2-(1-(4- methoxybenzoyl)-3-methyl-5-oxo-1H-pyrazol-4(5H)- ylidene)hydrazinyl)-N-(pyrimidin-2-yl)benzenesulfonamide. Example 3: Method of synthesis of derivative of formula (I-2) (E)-4-(2-(1-(4- fluorobenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H-pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide Step 1: Preparation of (E)-ethyl 3-oxo-2-(2-(4-(N-(pyrimidin 2yl) sulfamoyl) phenyl) hydrazono) butanoate (Intermediate-3): a) Dissolving 100g of 0.4mol of 4-amino-N-(pyrimidin-2-yl) benzenesulfonamide in 700ml solution of hydrochloric acid and cool the mixture to 0-5° C. b) Dissolving 41g of sodium nitrite in 200 ml of water and dropwise adding the solution into the mixture of step (a). c) Stirring the mixture of step (b) at 0-5° C for 30 minutes. d) Dissolving 238g of sodium acetate in 600ml of water and stir to form a mixture. e) Dissolving 54.6g of 0.42mol ethyl acetoacetate to the solution of step (d) and maintaining the mixture at 0-5° C. f) Dropwise adding the mixture of step (c) into the mixture of step (e) while keeping the temperature 0-5° C within 30minutes. g) Stirring the reaction mixture of step (f) for 30 minutes. h) Filtering the reaction mixture of step (g) and washing it with 300ml of cold water. i) Drying the compound under vacuum at 45-50°C to get Intermediate (3). Step 2: Preparation of (E)-4-(2-(1-(4-fluorobenzoyl)-3-methyl-5-oxo-1,5-dihydro- 4H-pyrazol-4-ylidene)hydrazinyl)-N-(pyrimidin-2-yl)benzenesu lfonamide (Formula I-2): a) Dissolving 100.0g of Intermediate 3 and 40.0g of 4- fluorobenzohydrazide into 100ml of acetic acid at room temperature. b) Stirring the reaction mixture of step (a) at 25-30 0 C for 30 minutes and then heat it at 95-100 0 C under stirring for 40 hours. c) Cooling the mixture of step (b) to 35-40 0 C and keep stirring at the same temperature for 1 hour. d) Filter the precipitated compound of step (c) and washing it with 200 ml of cold water. e) Drying the compound of step (d) under vacuum at 45-50 0 C to get 20.0g of the crude compound. f) Suspending 20.0g of the crude compound of step (e) in 1000ml of methanol and reflux the mixture under stirring for 2-3 hours. g) Cool the suspension of step (f) to 25-30 0 C and is stirred for 1 hour. h) Filtering the compound of step (g) and washing it with 200ml of methanol and drying the compound under vacuum at 45-500C to get the pure compound with formula (I-2) (E)-4-(2-(1-(4- fluorobenzoyl)-3-methyl-5-oxo-1,5-dihydro- 4H-pyrazol-4- ylidene)hydrazinyl)-N-(pyrimidin-2-yl)benzenesulfonamide. Example 4: Analysis of the derivatives a) (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H-pyrazol-4- ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide (Formula I-1): The chemical formula of the compound (E)-4-(2-(1-(4-methoxybenzoyl)-3-methyl- 5-oxo-1, 5-dihydro-4H-pyrazol-4-ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide is C22H19N7O5S having the molecular weight 493.50. Figure 1 illustrates FT-IR results, Figure 3 illustrates the Mass spec and Figure 5 illustrates NMR results of Formula (I-1). b) (E)-4-(2-(1-(4-fluorobenzoyl)-3-methyl-5-oxo-1, 5-dihydro-4H-pyrazol-4- ylidene) hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide (Formula I-2): The chemical formula of (E)-4-(2-(1-(4-fluorobenzoyl)-3-methyl-5-oxo-1, 5- dihydro-4H-pyrazol-4-ylidene)hydrazinyl)-N-(pyrimidin-2-yl) benzenesulfonamide is C 21 H 16 FN 7 O 4 S having molecular weight 481.46. Figure 2 illustrates FT-IR results, Figure 4 illustrates the Mass spec and Figure 6 illustrates NMR results of Formula (I-2). Example 5: Toxicity Studies of the Derivatives a) Toxicity Prediction for derivative of Formula (I-1) Result: Based on the available results and toxicity prediction from the QSAR software, it is concluded that the compound is non-hepatotoxic, noncarcinogenic, non-immunotoxic, non-mutagenic and noncytotoxic with a probability of 0.51, 0.51, 0.96, 0.74 and 0.82, respectively. Based on the predicted LD50 (4000 mg/kg b.wt.) and class (class 5), it is concluded that the safety index of the compound is very wide and compound is expected to be nontoxicity under the conditions and procedures followed in the present QSAR study. b) Toxicity Prediction for derivative of Formula (I-2) Result: Based on the available results and toxicity prediction from the QSAR software, it is concluded that the compound is non-hepatotoxic, noncarcinogenic, non-immunotoxic and mutagenicity with a probability of 0.51, 0.57, 0.98, 0.82 and 0.69, respectively. Based on the predicted LD50 (4000 mg/kg b.wt.) and class (class 5), it is concluded that the safety index of the compound is very wide and compound is expected to be nontoxicity under the conditions and procedures followed in the present QSAR study. Example 6: Biological Evaluation Antifungal activity: The biological activity such as antifungal activity of compound of formula (I-1) & compound of formula (I-2) were tested on a potato dextrose agar (PDA) medium on each of these vegetable pathogenic strains. The fungicidal activity of compound of formula (I-1) & compound of formula (I-2) were studied at 1000 ppm concentration in vitro. Plant pathogenic organisms used were Aspergillus Niger, Nigrospora Species, Rhizopus Nigricum, Botrydepladia Theobromae and Fusarium Oxyporium. Such a Potato agar medium contained dextrose 20g, potato 200g, agar 20g and water 10 ml. The Compounds to be tested were hovering (1000 ppm) in a PDA medium and autoclaved at 100 °C for 14 minutes at 12 atm. pressure. Old cultures were employed five or more days. These Potato agar media were mixed into sterile Petri plates and the microorganisms were immunized after cooling the Petri plates. The % zonal inhibition calculated for fungi was after five days using the formula given below: % of inhibition = 100 (X-Y) / X Where, Y = test plate Area of colony X = control plate Area of colony Table 1: Antifungal activity of compound of formula (I-1) and compound of formula (I-2) Antibacterial activity: It was mixed with 0.3-0.8 ml of 48 hour mixed well and old culture especially by normal stirring before adding on the Petri dish sterilized (50 ml in each dish). Nutrient agar broth was added in an aqueous beaker and boiled to 55 °C with occasional shaking to form well mixing. The melted mass was allowed to mix (1.5 hour) and after the mixing the “cups” were made by pressing into the agar with spooning out the pressed part of agar and sterile cork borer. Into this cups 0.10 ml of prepared test solution (prepared by dissolving 10.0 gm of sample in 10 ml Dimethyl formamide) was added by micropipette which was sterile. The plates were noted by appropriate codes. Table 2: Antibacterial activity of compound of formula (I-1) and compound of formula (I-2)