FACILE SYNTHESIS OF IN-VIVO INSECTICIDAL AND ANTIMICROBIAL EVALUATION OF BIS HETEROCYCLIC MOIETY FROM PET WASTE

Recycling of Plastic Solid Waste (PSW) using green tool such as solar energy which used for degradation of plastic solid bottles (PET) to afford terepthalic acid which estrified using butanol/H2SO4 to afford dibutyl terephthalate as a starting material which has been chemically modified to yield new bis (1,2,4-triazole-3thiol) 5, 1,3,4-oxadiazole 6, 8, 15, 1,3-thiazolidine 9, 1 H-pyrazole 10, 11 and 13 derivatives. Interestingly all the synthesized compounds exhibited good in-vivo insecticidal activity against the Cluex pipiens and musca domestica and also they were assayed to have high antimicrobial activity.


Chemical Protocols
All melting points were measured on a Gallenkamp melting point apparatus and uncorrected. The infrared spectra were recorded in potassium bromide disks on a Pye Unicam SP-3-300 and Shimadzu FT IR 8101 PC infrared spectrophotometers. The NMR spectra were recorded on a Varian Mercury VX-500 NMR spectrometer. 1 HNMR spectra were run at 500 MHz and 13 CNMR spectra were run at 75.46 MHz in Dimethyl Sulphoxide (DMSO-d 6 ). Chemical shifts were related that of the solvents. Mass spectra were recorded on a Shimadzu GCMS-QP-1000EX mass spectrometer at 70 e.v.

Prepartion of Terephthalic Dihydrazide (3) Via Hydrolysis of Polyethylene Terepthalate (PET)
Plastic bottles (22 g) were cut into a small strips and mixed with 50% NaOH solution placed in sun light for 5 weeks to obtain the sodium salt of terephthalte and then dissolved in water then acidify by (5 mol L −1 ) H 2 SO 4 to afford white precipitate of terephthalic acid (1), yield 90%; m.p. above 300°C. Then, terephthalic acid (10 g, 1.66 moL) was refluxed in absolute butanol (30 mL) and H 2 SO 4 (5 mL) for 6h to get dibutyl terephthalate (2). A mixture of 2 (10 mL) and hydrazine hydrate (6.0 mL, 99%) in 50 mL of absolute ethanol was refluxed for 5 h. The reaction mixture was left to cool and the separated solid was filtered off and recrystallized from EtOH/DMF to give terephthalic dihydrazide (3) (Palekar et al., 2009).

Synthesis of N' 1 , N' 4 -Dibenzoyl Terephthalo Hydrazide (14)
A mixture of the hydrazide 3 (2 mmoL) and benzoyl chloride (2 mmoL) in pyridine (20 mL) was refluxed for 24 h, then the reaction mixture was then cooled, diluted with water and acidified with dil HCl. The resulting solid was filtered, washed with water, dried and recrystallized from ethanol/DMF to afford 14; yield 65% mp: 185°C; IR (KBr) v max /cm

Insecticidal Activity
Tested insects: Culex pipiens (Culicidae: Diptera) and Musca domestica (Mucidae: Diptera). Larvae of Culex pipiens and Musca domestica were provided from Medical Entomology Institute and transferred to the laboratory of Entomology Department-Faculty of Science-Ain Shams University where self-perpetuating colonies were established and maintained during the present study as described by (Kamel et al., 2005) for Culex pipiens. Preliminary, toxicological bioassay tests were carried out on tested compounds as a modification for the described method. Mortality data was analyzed by using log-probit analysis to estimate probity regression line and calculate LC 50 , LC 95 , slope function by applying the computer program (Chornous et al., 2005). If the control mortality was between 5 and 20%, the percentage mortalities were corrected by Abbott's formula (Wright, 1971;Abbott, 1925;Chornous et al., 2005). Ten tested compounds were bioassay against the 3rd instars of the Cluex pipiens larvae and musca domestica larvae in the laboratory (Table 1 and 2). Which were statistically calculated for LC 50 and LC 95 at p = 0.05. The tested compounds showed different toxicity against them ( Fig. 1-4). All the tested compounds showed a certain effect on Culex pipiens and Musca domestica as illustrated in (Table 1 and 2) which clear that 1,3,4-oxadiazol derivative 16 gave higher more potent activity than open chain compound N' 1 ,N' 4 -dibenzoyl terephthalo hydrazide (14) Despite of other derivatives were closely resulted in potency which were found to possess crossing regression lines of nearly equal slope values. This may suggest that these compounds have different mode of action against the tested insect larvae and homogenous effect on the population (Busvine, 1971). Also compound 1H-pyrazol derivative 10 exhibited high activity in action of Culex pipiens rather than Musca domestica. In general mosquitoes were more susceptible than housefly to tested compounds. The susceptibility of Culexpipiens and Musca domestica larvae to a pyrethroid insecticide (Deltamethrine) and an insect growth regulator (Flufenoxuron) to compare them with the tested compounds. The results presented in Table 3 and illustrated in Fig. 5 which shows that Culexpipiens and Muscadomestica larvae are generally most susceptible to Deltamethrine insecticide then Flufenoxuron at the basis of LC 50 values.
Although decreasing potency of the newly synthesized compounds in comparing with traditional insecticides, the new compound 15 is still promising because of it was produced from waste material and needs more investigations to detect its mode of action: • Lines 1 and 2 represent Deltamethrine for Muscadomestica and Culexpipiens respectively • Lines 3 and 4 represent Flufenoxuron for Musca domestica and Culex pipiens respectively

Antimicrobial Activity
Ten of the newly synthesized target compounds were evaluated for S. pneumonia, S. aureus (B), S. typhimurium and E.coli (30) for Gram+ve and -ve Bacteria, respectively and antifungal activites for C. albicans (A) and A. flavu. The results were recorded for each of the tested compounds as the average diameter of the Inhibition Zone (IZ) of bacterial or fungal growth around the disks in mm. The results, depicted in Table 4 revealed that the most of the tested compounds displayed variable inhibitory effects on the growth of G+ and G-bacterial strain and antifungal strains. In general, most of the tested compound 1,2,4 triazole derivative 5 exhibited high degree of inhibition against all types of strains. Agar well diffusion method showing Antimicrobial activities of the tested compounds compared with reference (µm) and also compound 1,3,4-oxadiazol-2-amine derivative 6 has high activity against all strains except S.pneumonia (s), These bulky substituent deteriorate the antibacterial and antifungal activity of these compounds (8,9,14,12,15 and 10) show a moderate activity against bacteria and fungi strains ( Table 5 and 6) and shows the inhibition zone in (Fig 6-8).  Table 3. Larval mortality and LC 50 and LC 95 values for the Cluex pipiens and Musca domestica larvae exposed to Deltamethrine     Table 4. Agar well diffusion method showing Antimicrobial activities of tested compounds compared with reference (mm) and also compound 1,3,4-oxadiazol-2-amine derivative 6 has high activity against all strains except S. pneumonia (s), These bulky substituent deteriorate the antibacterial and antifungal activity of these compound (8,9,14,12,16 and 10) show a moderate activity against bacteria and fungi strains Table 4.

OJBS
Also the MIC activity were assayed in Table 5 and 6 and it was showed that the 1,2,4 triazole derivatives 5 exhibited resemble activity such the reference drug against S. typhimurium (G+) against E. coli by measuring theaverage diameter of the Inhibition Zone (IZ):

DISCUSSION
Degradation of Poly Ethylene Terephthalate (PET) utilizing Sun energy as a source of heat to afford terephthalic acid (1) which was esterified to dibutyl terephthalate (2) which was treated with hydrazine hydrate to afford terephthalic dihydrazide (3). Treatment of (3) with phenylisothiocynate in refluxing benzene gave the bisthiosemicarbazide 4 (Shaker et al., 2005;Pradip, 2008;Ram and Vlietinck, 1988). Furthermore, the intramolecular cyclization of 4 takes place upon heating with KOH to produce the 1,2,4 triazole derivative 5. Its mass spectrum revealed a peak corresponding to the molecular ion at m/z 428(M + ) and its IR spectrum was free of NH and C = O absorption bands. Interestingly, when bis-thiosemicarbazide 4 was boiled with potassium iodide and iodine in DMF, it gave 1,3,4-oxadiazol-2-amine derivative 6 ( Fig. 9) which was confirmed on the basis of its elemental analysis and spectral data; (cf. experimental).
The required bis-dithiocarbazinate 7 was synthesized by reacting dihydrazide 3 with carbon disulfide and potassium hydroxide in ethanol. This salt underwent ring closure with an ethyl 2-bromoacetate to give bis-1,3,4-oxadiazole derivative 8. The structure was confirmed by IR, 1 H NMR and mass spectra. For example, the 1 H NMR of 8 showed a triplet signal at δ 1.18 ppm due to methyl group and singlet signal at 4.16 due to active methylene, whereas its mass spectrum revealed molecular ion peak at m/z 450 (M + ). While the reaction of the salt 7 with 1-phenyl-2-bromoethanone yilded 1,3-thiazolidine derivative 9 which was confirmed by spectral data (Fig. 10). For example IR spectrum of 9 showed a strong absorption band due to NH of Pyrazole at 3057cm −1 , its 1 H NMR spectrum showed a singlet at δ 4.25 (NH, D 2 O exchangable) and its mass spectrum showed a peak corresponding to its molecular ion at m/z 362 (M + ).

OJBS
The structure of 16 was confirmed by the spectral data, For example its IR spectrum showed the absence of the absorbtion bands of NH and C = O groups, while 1 H NMR spectrum of 15 reaveled multiplet at δ 8.21 due to the aromatic protons (Fig. 13).

CONCLUSION
PET waste was utilized as chemical feed stock for preparing new cheap compounds. Thus, the preparation of the novel compounds 5, 6, 8, 9, 10, 11, 13 and 15 were carried out starting from terepthalic dihydrazide which is being obtained from PET waste. Insecticidal and antimicrobial activities of the synthesized heterocycles were studied and we concluded that 1,3,4oxadiazol derivative 16 displayed an insecticidal activity against Cluexpipiens larvae and muscadomestica larvae; where the most promising 1,2,4-triazole derivative 5 show high activity in all strain media comparable with to commercial compounds.

ACKNOWLEDGMENT
The researchers express their thanks to Omnia Mohamed. Kamel for the Biological part.