There is restriction of free rotation ( freedom) across. for vicinal hydrogen & OH to take trans position.

Transcription

1 5. SUMMARY & CONCLUSION Synthesis of Novel dihydro methyl Chalcones and their biological activity. The starting materials for the synthesis of various substituted dihydro 3-methyl chalcones were substituted benzaldehyde and 2-hydroxypropiophenone. The reactions were based on Claisen- Schmidt condensation but without dehydration process. An attempt has been made to synthesized 15 novel dihydro chalcones which is differ from classical Claisen-Schmidt condensation would have been explained as follows. In an attempt to modify the classical Claisen-Schmidt synthesis for chalcones which involves aldol condensation followed by dehydration. A strategy was envisaged after proper study based on Prentic Hall Frame Work Molecular Model that instead of acetophenone, if propiophenones are taken the OH (Hydroxy group) generated from CHO of aromatic aldehyde would prefer to be trans to the methyl with the results that no vicinal trans hydrogen atom will be available for the elimination of a water molecule. There is restriction of free rotation ( freedom) across. for vicinal hydrogen & OH to take trans position. O C H C CH 3 bond to make it difficult The yield of the synthesized compounds was in the range of 70-90%. All the synthesized compounds were purified by melting point determination, Rf value analysis, and the structures were characterized and established by U.V. IR, 1 H-NMR, 13 C-NMR and Mass spectrometry. All the physical data of the synthesized compounds like melting points, Rf value, yield, λ max, and partition coefficient has been determined given in table no; log P value for all the test compounds were determined by classical shake flask method using n- octanol and phosphate buffer of ph 7.4. All the test compounds have shown their log P values in the range of to λmax of all the test compounds were determined by UV spestrophotometer in HPLC grade methanol School of Pharmaceutical Sciences, Shoolini University. 206

2 Antioxidant Activity All the synthesized compounds (KLD-1 to KLD-15) were screened for their antioxidant activity. They were screened by DPPH free radical scavenging, and ABTS scavenging methods. The IC 50 values of all the synthesized compounds were determined and compared with the standard molecules (Ascorbic Acid). Most of the compounds were found to be moderately active while some of the compounds have shown excellent antioxidant activity like KLD-10, KLD-15, KLD-7 and KLD-4. The IC 50 values of these compounds were 8.3, and 12.8 µg/ml respectively by DPPH and 19.5, 19.7, 20.1 and 29.8 µg/ml respectively by ABTS method. Cytotoxic Activity All the synthesized compounds (KLD-1 to KLD-15) were evaluated for their cytotoxic activities in different human cancer cell lines, normal cell lines and standard drug molecule (Doxorubicin). Cell Lines; MCF-7 (Esterogenic positive), MDAMB-231, (Esterogenic negative). A-549 (Lung Cancer), PC-3 (Prostate cancer), Vero (Normal kidney epithelial cell line) Most of the compounds were moderately active; whereas some of the compounds were showed excellent cytotoxic activity. The compound KLD-4, KLD-7, KLD- 10 and KLD-15 have shown remarkably good broad spectrum cytotoxic activities, ranges from 3.0 to 6.5 µm against all the cell lines under study and compared with the standard compounds Doxorubicin. Antimicrobial Activity All the test compounds (KLD-1 to KLD-15) were screened for antimicrobial activity against six bacterial and two fungal strains namely Escherichia coli, Pseudomonas aeruginosa, Klebsiella-pneumonia, Staphylococcus aureus, Bacillus subtilis, and Bacillus cereus and two fungal strain Candida albicans, Aspergillus nigrum The standard drug were Amikicin, Gatifloxacin. The compound KLD-7 and KLD-15 showed excelent broad spectrum antimicrobial activity ranges from 4.1 to 8.3µg/ml as structural requirement is completly satisfied with the literature (Awasthi et al., 2009). KLD-5 and KLD-3 School of Pharmaceutical Sciences, Shoolini University. 207

3 have shown good antimicrobial activity ranges from 4.9 to 14.5 µg/ml. The remaining compounds have shown moderate activity Synthesis of trans chalcones and their biological activities. An attempt has been made to synthesized 15 trans chalcones which is differing from classical Claisen-Schmidt condensation would have been explained as follows. Specification; On the basis of different reaction conditions and spectroscopic data, it has been found that the compounds synthesized were categorized into two groups. The suggested compounds were trans at room temperature, whereas some compounds were only synthesized after heating under reflux. Under these conditions the α hydrogen to the carbonyl group is under magnetic an isotropic environment and is therefore extra ordinary downfield. The 1 HNMR data shows that different configuration of these compounds and chemical shift of α and β hydrogen atoms are very close to each other which is un usual, but could be a special case of protons which are chemically different but magnetically very close. All the synthesized compounds were purified by melting point (uncorrected) determination, Rf value analysis, and the structure were characterized and established by U.V. IR, 1 H-NMR and 13 C-NMR and Mass spectrometry. The yield of the synthesized compounds were in the range of 65-90% All the physical data of the synthesized compounds like melting points (uncorrected), Rf value, yield, λ max, and partition coefficient has been determined and summarized. All the synthesized compounds were evaluated for the antioxidant, antibacterial antifungal, and cytotoxic activity. Antioxidant Activity All the synthesized compounds (CASD-1 to CASD-15) were screened for their antioxidant activity. They were screened by DPPH free radical scavenging, and ABTS scavenging methods. The IC 50 of all the synthesized compounds were determined and compared with standard molecules, (Ascorbic Acid) enlisted in table no; School of Pharmaceutical Sciences, Shoolini University. 208

4 Most of the compounds were found to be good to moderately active while some of the compounds have shown excellent antioxidant activity like CASD-7 CASD-9, CASD-13, and CASD-8 The IC 50 value of these compounds were 8.7, and 12.4 µg/ml respectively by DPPH and 21.5, 29.2, 26.5 and 31.8 µg/ml respectively by ABTS method. Cytotoxic Activity All the synthesized compounds (CASD-1 to CASD-15were evaluated for their cytotoxic activities in different human cancer cell lines, normal cell lines and standard drug molecule (Doxorubicin). Cell Lines; MCF-7 (Esterogenic positive), MDAMB-231, (Esterogenic negative). A-549 (Lung Cancer), PC-3 (Prostate cancer), Vero (Normal kidney epithelial cell line) Most of the compounds were moderately active; whereas some of the compounds have shown excellent cytotoxic activity. The compounds like CASD-5, CASD-7, CASD--10 and CASD--13 shown remarkably good broad spectrum cytotoxic activities, ranging 3.3 to 6.5 µm against all the cell lines under study, while standard drug Doxorubicin showed activities between 2.2 to 4.7 µm. Antimicrobial Activity All the test compounds (CASD-1 to CASD-15) were screened for antimicrobial activity against six bacterial and two fungal strains namely Escherichia coli, Pseudomonas aeruginosa, Klebsiella-pneumonia, Staphylococcus aureus, Bacillus subtilis, and Bacillus cereus and two fungal strain Candida albicans, Aspergillus nigrum The compound CASD-10, CASD-8, and CASD-3 showed remarkaly good broad spectrum antimicrobial activity ranges from 4.9 to 7.5 µg/ml, 5.3 to 9.8 µg/ml and 7.9 to 12.4 µg/ml respectively as structural requirement is completly satisfied with the literature (Dhar et al.,2010; Jayshree et al.,2012) The remaining compounds have shown moderate activity. School of Pharmaceutical Sciences, Shoolini University. 209

5 5.3. Synthesis of bioactive chalcones and their dihydro pyrazole derivatives with their biological activity. An attempt has been made to synthesized 15 chalcones which were further subjected to react with hydrazine hydrate to produce 15 dihydro pyrazole chalcone analogues. All the synthesized compounds were purified by melting point determination, Rf value analysis, and the structure were characterized and established by U.V. IR, 1 H-NMR and 13 C-NMR and Mass spectrometry. The yield of the synthesized compounds were in the range of 70-90% All the physical data of the synthesized compounds like melting points (uncorrected), Rf value, yield, λ max, and partition coefficient has been determined and summarized in respective table. All the synthesized compounds were evaluated for the antioxidant, antimicrobial, and cytotoxic activity. Antioxidant Activity All the synthesized compounds (JAD-1 to JAD-15) were screened for their antioxidant activity. They were screened by DPPH free radical scavenging, and ABTS scavenging methods. The IC 50 of all the synthesized compounds and the standard molecules, (Ascorbic Acid) were determined and enlisted in table no; Most of the compounds were found to be good to moderately active while some of the compounds have shown excellent antioxidant activity like JAD-3 (7.3 µg/ml DPPH and 25.9 µg/ml ABTS), JAD -5, (10.2 µg/ml DPPH and 26.2 µg/ml ABTS The compounds like JAD-2, JAD-6, JAD-14 and JAD-15 have shown good activity ranges from 13.7 µg/ml to 18.4 µg/ml by DPPH method and 23.2 µg/ml to 31.3 µg/ml by ABTS method. Cytotoxic Activity All the synthesized compounds (JAD-1 to JAD-15) were evaluated for their cytotoxic activities in different human cancer cell lines, normal cell lines and standard molecule (Doxorubicin). Cell Lines; MCF-7 (Esterogenic positive), School of Pharmaceutical Sciences, Shoolini University. 210

6 MDAMB-231, (Esterogenic negative). A-549 (Lung Cancer), PC-3 (Prostate cancer), Vero (Normal kidney epithelial cell line) Most of the compounds were found to be moderately active; whereas some of the compounds showed excellent cytotoxic potential. The compounds like JAD-2, JAD-3, JAD-6 JAD-12 and JAD-15 have shown remarkably good broad spectrum cytotoxic activities, (IC 50 Value) ranging from 3.0 to 6.6 µm against all the cell lines under study, while standard drug Doxorubicin showed cytotoxic activities between 2.2 to 4.6 µm. Antimicrobial Activity. All the test compounds (JAD-1 to JAD-15) were screened for antimicrobial activity against six bacterial and two fungal strains namely Escherichia coli, Pseudomonas aeruginosa, Klebsiella-pneumonia, Staphylococcus aureus, Bacillus subtilis, and Bacillus cereus and two fungal strain Candida albicans, Aspergillus nigrum. Most of the compounds have shown moderate antimicrobial activity whereas some of the compounds have shown excellent antimicrobial activity. The JAD-1, 9, 13, and 14 showed excelent broad spectrum antimicrobial activities between 4.2 to 7.6 µg/ml against all microorganism Out come and Scope. It has been concluded that all the isolated compounds, their analogues and synthetic compounds were well characterized and evaluated (in vitro) for their biological activities. Isoflavones were showing excellent cytotoxic activity, when interact with estrogenic receptors, hence could be good anticancer agents, especially for breast cancer and prostate cancer. In vivo analyses of all natural, semisynthetic and synthetic compounds are required to achieve the scientific goal. The recent report available on analogues of isoflavones for their farnesyltransferase inhibitors will initiate our study to isolate/synthesize many more of such substituted isoflavones.. There is a future scope for the isoflavone to be targeted for newer targets such as CYP-450 aromatase to explore their anticancer agents. Some of the isoflavone product have recently launched. Eg. Ipriflavone a marketed isoflavone reduces total LDL cholesterol level. School of Pharmaceutical Sciences, Shoolini University. 211

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