- Email: [email protected]
Evaluation of interactions between metformin and cytotoxic drugs in HEPG2 cells
Abstracts / Toxicology Letters 258S (2016) S62–S324
aim of this study was to determine the association between the maternal MDR1 C1236T polymorphism and mercury levels in a triad of mother–placenta–fetus. The study population comprised 110 mothers and their respective placentas and umbilical cords. Venous blood samples from mothers were collected to investigate both MDR1 C1236T polymorphism and mercury levels. Placentas and cord blood samples were collected to measure mercury levels. MDR1 C1236T polymorphism was detected standard PCR–RFLP technique. Mercury levels were quantified by cold vapor atomic absorption spectrometry (CVAAS). The mean mercury levels of maternal blood, placenta and cord blood were 3.28 ± 1.38 g/L, 12.43 ± 11.38 g/kg and 3.65 ± 3.73 g/L, respectively. The MDR1 C1236T genotype frequencies of mothers were 30% homozygote typical (CC), 50.91% heterozygote (CT) and 19.09% homozygote atypical (TT). Our study results showed that maternal blood and placental mercury levels were lower in mothers with TT genotype than those with CC and CT genotypes, whereas cord blood mercury concentrations were higher in mothers with TT genotype, but these results were not statistically significant (p > 0.05). The present study indicated that MDR1 C1236T polymorphism may have an effect on mercury levels of mother–placenta–fetus. We believe that further studies with larger study subjects are needed to prove this hypothesis. This study is supported by the Ankara University Scientific Research Projects Coordination Unit (BAP; Project Number: 15B0217001). http://dx.doi.org/10.1016/j.toxlet.2016.06.1732 P10-066 Toxicity of aerosols of propylene glycol, vegetable glycerin and nicotine in Sprague-Dawley rats in a 90-d OECD 413 sub-chronic inhalation study B. Phillips 1,∗ , D. Sharma 1 , D. Sciuscio 2 , E. Veljkovic 1 , S. Lebrun 2 , J. Verbeeck 2 , W. Tan 1 , U. Kogel 2 , J. Ho 1 , G. Vuillaume 2 , P. Leroy 2 , J. Hoeng 2 , M.C. Peitsch 2 , P. Vanscheeuwijck 2 1
Philip Morris International Research Laboratories Pte. Ltd., 50 Science Park Road, Singapore3 2 Philip Morris International R&D, Philip Morris Products S.A., Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland3 The toxicity of three major E-cigarette liquid constituents; propylene glycol (PG), vegetable glycerin (VG) and nicotine, were characterized in a 90-day sub-chronic inhalation study according to the OECD 413 testing guideline. Sprague-Dawley rats were exposed 6 h/day, 5 d/week to filtered air, saline, to 3 test atmosphere concentrations of propylene glycol and vegetable glycerin (0.17/0.21 mg/llow, 0.52/0.63 mg/l-medium, 1.52/1.89 mg/l-high) or to the same PG/VG aerosols containing nicotine (fixed at 0.023 mg/l). The evaluation of all experimental endpoints revealed findings mainly associated with nicotine exposure. Rats exposed to the nicotinecontaining aerosols had gender-specific changes to body weight – lower in males, higher in females. Clinical chemistry evaluation revealed a slight increase in liver enzyme activity, and decreased concentrations of metabolic markers (glucose, cholesterol). Organ weight changes included increased liver and adrenal weight, and lower thymus weights. No significant changes were detected in the numbers of inflammatory cells or excreted pro-inflammatory cytokines in the bronchoalveolar lavage fluid of animals exposed to any of the test aerosols, which was confirmed by minimal
3
Part of Philip Morris International Group of Companies.
S201
histopathological changes to the respiratory tract following exposure. Similarly, rare microscopic findings in non-respiratory tract organs in all exposure groups were considered incidental, except for mild vacuolization in the liver in the nicotine-exposed animals. In summary, these results indicate that the inhalation of an aerosol generated from nebulized mixture of nicotine, PG, and VG caused minimal toxic effects of low severity, mostly attributed to nicotine and with no combinational effect of PG and VG. http://dx.doi.org/10.1016/j.toxlet.2016.06.1733 P10-067 Comparison of prenatal developmental toxicity of Pentapropenylsuccinate monoethanolamide and Pentapropenylsuccinimido-hexanoate M. Lee ∗ , J. Bietz, R. Kreiling Clariant Produkte (Deutschland) GmbH, Germany Pentapropenylsuccinate monoethanolamide (PSA-EA) and Pentapropenylsuccinimido-hexanoate (PSI-HA) are industrial chemicals. They are manufactured by using Pentapropenylsuccinic anhydride as precursor and thus share identical lipophilic moiety, but they differ in that PSA-EA is a succinic acid half amide while PSI-HA is a succinimide. On the basis of chemical structure analysis, no endpoint of concern was identified for PSA-EA. But for PSI-HA, the developmental toxicity was found critical, due to the structural similarity to some well-known teratogenic “cyclic imides”. Additionally, increases of stillbirth and postnatal mortality were found in rats treated with PSI-HA in reproductive/developmental toxicity screening test. Pregnant rats were treated with PSA-EA or PSI-HA during gestation days 5–19. General behavior, body weight and food consumption were measured. At scheduled Caesarian section, dams were evaluated for body weight, gravid uterine weight and status of implantation sites. Fetuses were examined for external, visceral, and skeletal abnormalities. Head sections were performed when necessary. In rats treated with PSA-EA up to dose of 1000 mg/kg bw, no maternal and developmental toxicity was found. In rats treated with PSI-HA at doses of 200, 40 and 8 mg/kg bw, no maternal toxicity and no effect on the gestational parameters were found. However, changes of nasopharynx and incompletely fused nasal septa to the palate were noted at 200 mg/kg bw and increased incidences of small spleen were found down to 8 mg/kg bw. Although structurally related, PSA-EA and PSI-HA are not comparable with respect to developmental toxicity. The experimental data support PSA-EA is not a developmental toxicant. http://dx.doi.org/10.1016/j.toxlet.2016.06.1734 P10-068 Evaluation of interactions between metformin and cytotoxic drugs in HEPG2 cells S. Sabuncuoglu ∗ , Z. Ünal Hacettepe University, Faculty of Pharmacy, Department of Toxicology, Turkey Metformin (1,1-dimethylbiguanidehydrochloride, MET) is an oral biguanide antidiabetic drug that is widely used for the treatment of type 2 diabetes mellitus. Recent studies have shown that MET has also different biological activities such as anticancer,
S202
Abstracts / Toxicology Letters 258S (2016) S62–S324
cardioprotective, anti-aging. Type 2 diabetes mellitus patients are at increased risk of many forms of malignancies. Unfortunately, there is limited research on possible interactions between antidiabetic drugs and anticancer agents. The present study investigates possible interactions between metformin (MET) and doxorubicin (DOX), cisplatin (CIS), and gemcitabine (GEM). For this purpose, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT which is a colorimetric assay was performed for assessing cell metabolic activity hence cell viability. MET has been applied as alone or in combinations with DOX, CIS and GEM on HepG2 cell line for 24, 48 and 72 h at different doses. DOX has synergistic effect in combination with MET at different doses in 24, 48 and 72 h. On the other hand, CIS showed synergistic and antagonistic effects at different concentrations in both 24 and 72 h but not in 48 h. In addition, there is also a synergistic effect at low doses of GEM combined with MET in 24 and 72 h. In conclusion, use of MET can change the efficacy of chemotherapeutic drugs. Thus, the diabetic cancer patients who take MET has to be considered in terms of drug interactions. http://dx.doi.org/10.1016/j.toxlet.2016.06.1735 P10-070 Effect of metallothionein polymorphism on blood, hair and urinary mercury levels in Turkish women D. Kaya Akyüzlü 1 , Z. Kayaaltı 1,∗ , F. Özdemir 1 , B. Yüksel 2 , T. Söylemezo˘glu 1 1
Ankara University, Institute of Forensic Sciences, Ankara, Turkey Turkish National Police, Ankara Police Forensic Laboratory, Ankara, Turkey 2
Metallothioneins (MTs) are bivalent metal binding-proteins that present in virtually all living organisms. They are also considered as an important defense protein for the detoxification of metals. The aim of this study was to determine the effect of metallothionein polymorphism on blood, hair and urinary mercury levels in 93 Turkish women. MT2A-5 A/G polymorphism was detected standard PCR–RFLP technique. Mercury levels of blood, hair and urine of women were quantified by cold vapor atomic absorption spectrometry (CVAAS). A total of 85 mothers were homozygote typical (AA) genotype and 8 mothers were heterozygote (AG) genotype for MT2A polymorphism. No homozygote atypical (GG) mothers were observed. The mean mercury levels of blood, hair and urine were 3.23 ± 1.46 g/L, 100.98 ± 173.10 g/kg, 2.83 ± 2.55 g/L, respectively. Blood, hair and urinary mercury levels were lower in Turkish women with AG genotype compared to AA genotype, but it was not statistically significant (p > 0.05). These results indicated that MT2A-5 A/G polymorphism may cause individual susceptibility in view of blood, hair and urinary mercury levels in Turkish women. However, further studies involving multiple centers and greater sample size are needed. This study is supported by the Ankara University Scientific Research Projects Coordination Unit (BAP; Project Number: 15B0217001). http://dx.doi.org/10.1016/j.toxlet.2016.06.1736
P10-071 Synthesis and in vitro antioxidant activity studies of melatonin analogue compounds S. Süzen 1,∗ , H. Shirinzadeh 2 , C. Karaaslan 1 , H.G. Orhan 3 , B. Eren 3 1
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry 06100 Tandogan, Ankara, Turkey 2 Erzincan University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Erzincan, Turkey 3 Ege University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, I˙ zmir, Turkey Recent research has proved that the indole ring in the melatonin (MLT) molecule is the reactive center dealing with oxidants due to its high resonance stability and very low activation energy barrier towards free radical reactions. Indole is found to reduce formation of reactive oxygen species and to scavenge hydroxyl radical directly. The aim of this study is to synthesize and examine possible in vitro antioxidant effects of indole-based MLT analogue compounds. As a part of our ongoing study nineteen indole hydrazide/hydrazone derivatives were synthesized, characterized and their in vitro antioxidant activity was investigated by three different assays: by evaluating their reducing effect against oxidation of a redox sensitive fluorescent probe, by examining their protective effect against H2 O2 -induced membrane lipid peroxidation and by determining their inhibitory effect on AAPH-induced hemolysis of human erythrocytes. The results of three in vitro antioxidant experiments indicated significant antioxidant activity for most of the synthesized indole derivatives, even higher than MLT itself, revealing differences in their relative potencies probably related to electronic distribution. This work was supported by The Scientific and Technological Research Council of Turkey (TÜBI˙ TAK) Grant 109S099. http://dx.doi.org/10.1016/j.toxlet.2016.06.1737 P10-072 Indole-based melatonin analogue compounds as cellular ROS inhibitors: Investigation on the peroxidation of human erythrocyte membranes and oxidative hemolysis S. Süzen 1,∗ , Y. Ad 1 , C. Karaaslan 1 , H. Shirinzadeh 2 , T. Coban 3
1
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, 06100 Tandogan, Ankara, Turkey 2 Erzincan University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Erzincan, Turkey 3 Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, 06100 Tandogan, Ankara, Turkey
Melatonin (MLT), the main secretory product of the pineal gland is a well-known antioxidant and free radical scavenger. It scavenges a variety of reactive oxygen and nitrogen species including hydroxyl radical, hydrogen peroxide, singlet oxygen, nitric oxide and peroxynitrite anion. In this study new melatonin analogue compounds were synthesized and most of the synthesized compounds showed strong inhibitory effect on the superoxide radical scavenging assay at 1 mM concentration (79–95%). The active compounds displayed higher antioxidant activity then that of MLT. Compound 1p (5-chloro-1H-indole-3-carboxaldehyde (3,5-dichlorophenyl)hydrazone) demonstrated the best inhibitory activity with 95%. Almost all the tested compounds possessed strong scavenging activity against the DPPH radical scavenging activity with IC50 values (2–60 M). Compound that have anisic
aim of this study was to determine the association between the maternal MDR1 C1236T polymorphism and mercury levels in a triad of mother–placenta–fetus. The study population comprised 110 mothers and their respective placentas and umbilical cords. Venous blood samples from mothers were collected to investigate both MDR1 C1236T polymorphism and mercury levels. Placentas and cord blood samples were collected to measure mercury levels. MDR1 C1236T polymorphism was detected standard PCR–RFLP technique. Mercury levels were quantified by cold vapor atomic absorption spectrometry (CVAAS). The mean mercury levels of maternal blood, placenta and cord blood were 3.28 ± 1.38 g/L, 12.43 ± 11.38 g/kg and 3.65 ± 3.73 g/L, respectively. The MDR1 C1236T genotype frequencies of mothers were 30% homozygote typical (CC), 50.91% heterozygote (CT) and 19.09% homozygote atypical (TT). Our study results showed that maternal blood and placental mercury levels were lower in mothers with TT genotype than those with CC and CT genotypes, whereas cord blood mercury concentrations were higher in mothers with TT genotype, but these results were not statistically significant (p > 0.05). The present study indicated that MDR1 C1236T polymorphism may have an effect on mercury levels of mother–placenta–fetus. We believe that further studies with larger study subjects are needed to prove this hypothesis. This study is supported by the Ankara University Scientific Research Projects Coordination Unit (BAP; Project Number: 15B0217001). http://dx.doi.org/10.1016/j.toxlet.2016.06.1732 P10-066 Toxicity of aerosols of propylene glycol, vegetable glycerin and nicotine in Sprague-Dawley rats in a 90-d OECD 413 sub-chronic inhalation study B. Phillips 1,∗ , D. Sharma 1 , D. Sciuscio 2 , E. Veljkovic 1 , S. Lebrun 2 , J. Verbeeck 2 , W. Tan 1 , U. Kogel 2 , J. Ho 1 , G. Vuillaume 2 , P. Leroy 2 , J. Hoeng 2 , M.C. Peitsch 2 , P. Vanscheeuwijck 2 1
Philip Morris International Research Laboratories Pte. Ltd., 50 Science Park Road, Singapore3 2 Philip Morris International R&D, Philip Morris Products S.A., Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland3 The toxicity of three major E-cigarette liquid constituents; propylene glycol (PG), vegetable glycerin (VG) and nicotine, were characterized in a 90-day sub-chronic inhalation study according to the OECD 413 testing guideline. Sprague-Dawley rats were exposed 6 h/day, 5 d/week to filtered air, saline, to 3 test atmosphere concentrations of propylene glycol and vegetable glycerin (0.17/0.21 mg/llow, 0.52/0.63 mg/l-medium, 1.52/1.89 mg/l-high) or to the same PG/VG aerosols containing nicotine (fixed at 0.023 mg/l). The evaluation of all experimental endpoints revealed findings mainly associated with nicotine exposure. Rats exposed to the nicotinecontaining aerosols had gender-specific changes to body weight – lower in males, higher in females. Clinical chemistry evaluation revealed a slight increase in liver enzyme activity, and decreased concentrations of metabolic markers (glucose, cholesterol). Organ weight changes included increased liver and adrenal weight, and lower thymus weights. No significant changes were detected in the numbers of inflammatory cells or excreted pro-inflammatory cytokines in the bronchoalveolar lavage fluid of animals exposed to any of the test aerosols, which was confirmed by minimal
3
Part of Philip Morris International Group of Companies.
S201
histopathological changes to the respiratory tract following exposure. Similarly, rare microscopic findings in non-respiratory tract organs in all exposure groups were considered incidental, except for mild vacuolization in the liver in the nicotine-exposed animals. In summary, these results indicate that the inhalation of an aerosol generated from nebulized mixture of nicotine, PG, and VG caused minimal toxic effects of low severity, mostly attributed to nicotine and with no combinational effect of PG and VG. http://dx.doi.org/10.1016/j.toxlet.2016.06.1733 P10-067 Comparison of prenatal developmental toxicity of Pentapropenylsuccinate monoethanolamide and Pentapropenylsuccinimido-hexanoate M. Lee ∗ , J. Bietz, R. Kreiling Clariant Produkte (Deutschland) GmbH, Germany Pentapropenylsuccinate monoethanolamide (PSA-EA) and Pentapropenylsuccinimido-hexanoate (PSI-HA) are industrial chemicals. They are manufactured by using Pentapropenylsuccinic anhydride as precursor and thus share identical lipophilic moiety, but they differ in that PSA-EA is a succinic acid half amide while PSI-HA is a succinimide. On the basis of chemical structure analysis, no endpoint of concern was identified for PSA-EA. But for PSI-HA, the developmental toxicity was found critical, due to the structural similarity to some well-known teratogenic “cyclic imides”. Additionally, increases of stillbirth and postnatal mortality were found in rats treated with PSI-HA in reproductive/developmental toxicity screening test. Pregnant rats were treated with PSA-EA or PSI-HA during gestation days 5–19. General behavior, body weight and food consumption were measured. At scheduled Caesarian section, dams were evaluated for body weight, gravid uterine weight and status of implantation sites. Fetuses were examined for external, visceral, and skeletal abnormalities. Head sections were performed when necessary. In rats treated with PSA-EA up to dose of 1000 mg/kg bw, no maternal and developmental toxicity was found. In rats treated with PSI-HA at doses of 200, 40 and 8 mg/kg bw, no maternal toxicity and no effect on the gestational parameters were found. However, changes of nasopharynx and incompletely fused nasal septa to the palate were noted at 200 mg/kg bw and increased incidences of small spleen were found down to 8 mg/kg bw. Although structurally related, PSA-EA and PSI-HA are not comparable with respect to developmental toxicity. The experimental data support PSA-EA is not a developmental toxicant. http://dx.doi.org/10.1016/j.toxlet.2016.06.1734 P10-068 Evaluation of interactions between metformin and cytotoxic drugs in HEPG2 cells S. Sabuncuoglu ∗ , Z. Ünal Hacettepe University, Faculty of Pharmacy, Department of Toxicology, Turkey Metformin (1,1-dimethylbiguanidehydrochloride, MET) is an oral biguanide antidiabetic drug that is widely used for the treatment of type 2 diabetes mellitus. Recent studies have shown that MET has also different biological activities such as anticancer,
S202
Abstracts / Toxicology Letters 258S (2016) S62–S324
cardioprotective, anti-aging. Type 2 diabetes mellitus patients are at increased risk of many forms of malignancies. Unfortunately, there is limited research on possible interactions between antidiabetic drugs and anticancer agents. The present study investigates possible interactions between metformin (MET) and doxorubicin (DOX), cisplatin (CIS), and gemcitabine (GEM). For this purpose, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT which is a colorimetric assay was performed for assessing cell metabolic activity hence cell viability. MET has been applied as alone or in combinations with DOX, CIS and GEM on HepG2 cell line for 24, 48 and 72 h at different doses. DOX has synergistic effect in combination with MET at different doses in 24, 48 and 72 h. On the other hand, CIS showed synergistic and antagonistic effects at different concentrations in both 24 and 72 h but not in 48 h. In addition, there is also a synergistic effect at low doses of GEM combined with MET in 24 and 72 h. In conclusion, use of MET can change the efficacy of chemotherapeutic drugs. Thus, the diabetic cancer patients who take MET has to be considered in terms of drug interactions. http://dx.doi.org/10.1016/j.toxlet.2016.06.1735 P10-070 Effect of metallothionein polymorphism on blood, hair and urinary mercury levels in Turkish women D. Kaya Akyüzlü 1 , Z. Kayaaltı 1,∗ , F. Özdemir 1 , B. Yüksel 2 , T. Söylemezo˘glu 1 1
Ankara University, Institute of Forensic Sciences, Ankara, Turkey Turkish National Police, Ankara Police Forensic Laboratory, Ankara, Turkey 2
Metallothioneins (MTs) are bivalent metal binding-proteins that present in virtually all living organisms. They are also considered as an important defense protein for the detoxification of metals. The aim of this study was to determine the effect of metallothionein polymorphism on blood, hair and urinary mercury levels in 93 Turkish women. MT2A-5 A/G polymorphism was detected standard PCR–RFLP technique. Mercury levels of blood, hair and urine of women were quantified by cold vapor atomic absorption spectrometry (CVAAS). A total of 85 mothers were homozygote typical (AA) genotype and 8 mothers were heterozygote (AG) genotype for MT2A polymorphism. No homozygote atypical (GG) mothers were observed. The mean mercury levels of blood, hair and urine were 3.23 ± 1.46 g/L, 100.98 ± 173.10 g/kg, 2.83 ± 2.55 g/L, respectively. Blood, hair and urinary mercury levels were lower in Turkish women with AG genotype compared to AA genotype, but it was not statistically significant (p > 0.05). These results indicated that MT2A-5 A/G polymorphism may cause individual susceptibility in view of blood, hair and urinary mercury levels in Turkish women. However, further studies involving multiple centers and greater sample size are needed. This study is supported by the Ankara University Scientific Research Projects Coordination Unit (BAP; Project Number: 15B0217001). http://dx.doi.org/10.1016/j.toxlet.2016.06.1736
P10-071 Synthesis and in vitro antioxidant activity studies of melatonin analogue compounds S. Süzen 1,∗ , H. Shirinzadeh 2 , C. Karaaslan 1 , H.G. Orhan 3 , B. Eren 3 1
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry 06100 Tandogan, Ankara, Turkey 2 Erzincan University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Erzincan, Turkey 3 Ege University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, I˙ zmir, Turkey Recent research has proved that the indole ring in the melatonin (MLT) molecule is the reactive center dealing with oxidants due to its high resonance stability and very low activation energy barrier towards free radical reactions. Indole is found to reduce formation of reactive oxygen species and to scavenge hydroxyl radical directly. The aim of this study is to synthesize and examine possible in vitro antioxidant effects of indole-based MLT analogue compounds. As a part of our ongoing study nineteen indole hydrazide/hydrazone derivatives were synthesized, characterized and their in vitro antioxidant activity was investigated by three different assays: by evaluating their reducing effect against oxidation of a redox sensitive fluorescent probe, by examining their protective effect against H2 O2 -induced membrane lipid peroxidation and by determining their inhibitory effect on AAPH-induced hemolysis of human erythrocytes. The results of three in vitro antioxidant experiments indicated significant antioxidant activity for most of the synthesized indole derivatives, even higher than MLT itself, revealing differences in their relative potencies probably related to electronic distribution. This work was supported by The Scientific and Technological Research Council of Turkey (TÜBI˙ TAK) Grant 109S099. http://dx.doi.org/10.1016/j.toxlet.2016.06.1737 P10-072 Indole-based melatonin analogue compounds as cellular ROS inhibitors: Investigation on the peroxidation of human erythrocyte membranes and oxidative hemolysis S. Süzen 1,∗ , Y. Ad 1 , C. Karaaslan 1 , H. Shirinzadeh 2 , T. Coban 3
1
Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, 06100 Tandogan, Ankara, Turkey 2 Erzincan University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Erzincan, Turkey 3 Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, 06100 Tandogan, Ankara, Turkey
Melatonin (MLT), the main secretory product of the pineal gland is a well-known antioxidant and free radical scavenger. It scavenges a variety of reactive oxygen and nitrogen species including hydroxyl radical, hydrogen peroxide, singlet oxygen, nitric oxide and peroxynitrite anion. In this study new melatonin analogue compounds were synthesized and most of the synthesized compounds showed strong inhibitory effect on the superoxide radical scavenging assay at 1 mM concentration (79–95%). The active compounds displayed higher antioxidant activity then that of MLT. Compound 1p (5-chloro-1H-indole-3-carboxaldehyde (3,5-dichlorophenyl)hydrazone) demonstrated the best inhibitory activity with 95%. Almost all the tested compounds possessed strong scavenging activity against the DPPH radical scavenging activity with IC50 values (2–60 M). Compound that have anisic