ACCESS TO CONVENIENT SYNTHESIS OF OXAZINONE AND PYRIMIDINONE HETEROCYCLES DERIVED FROM 2(3H)- FURANONE

The conversion of the 2(3H)-furanone into the oxazinone and pyrimidinone derivatives are studied. A 2-(furan-2-ylmethylene)-4-oxo-4phenylbutanoyl azide 3 was synthesized to carry out these conversions through its thermolysis in dry benzene and base-catalyzed decomposition of this azide in the presence of different amines. The structures of all compounds were demonstrated from their spectral data and elemental analyses.


Experimental section General
Melting points were measured on Gallen Kamp electric melting point apparatus. The IR spectra were recorded using potassium bromide disks on Fourier transform infrared Thermo Electron Nicolet 7600 spectrometer (Thermo Fisher Scientific Inc., Waltham, MA, USA) at the Central Laboratory of Faculty of Science, Ain Shams University. The 1 H-NMR spectra were run at 400 MHz on a GEMINI 400 BB NMR spectrometer (GEMINI, Manufacturing & Engineering Inc., Anaheim, CA, USA) using tetramethyl silane (TMS) as an internal standard in deuterated dimethylsulphoxide (DMSO-d6) at the Main Defence Chemical Laboratory, Cairo. The mass spectra were recorded on Shimadzu GC-MS-QP-1000EX mass spectrometer (Shimadzu Scientific Instruments, Inc., USA) operating at 70 eV at the Microanalytical Center of Cairo University. The progress of all reactions was monitored by the thin layer chromatography using Merck Kiesel gel 60 F254 aluminum-backed plates.

Pyrolysis of azide 3
In 10 mL round-bottomed flask fitted with an air condenser was placed 0.5 g of the azide 3. The flask was heated on a sand bath whereby the azide was fused. Heating was continued for another one hour. The product obtained after recrystallization from petroleum ether, 80-100/ benzene, (2:1) was proved by direct comparison (mp., mixed mp. and TLC) to be the oxazinone 4.

Reaction of the azide 3 with benzylamine
Benzylamine (2 mmol) was added dropwise to a solution of azide 3 (2 mmol) in dry benzene (20 mL) and the mixture was stirred at room temperature for 2 h or heated under Scheme 1. Synthesis of compound 4.
The structure of 3 was inferred from its IR spectrum which displayed ν N3 of azide at 2142 cm -1 and devoid from NH & NH2 absorption bands.
Acyl azides are known to be suitable candidates for the synthesis of many acyclic and heterocyclic derivatives via their acid or base-catalyzed decompositions. Thus, it was of interest to the authors to construct oxazinone and pyrimidinone derivatives using the azide 3 as a key starting material. The reactions of 3 with benzylamine, hydrazine, and p-toluidine were studied (Scheme 2).
Thermolysis of the azide in dry benzene led to the construction of oxazinone derivative 4 as sole product (Scheme 1). The formation of 4 can be postulated via loss of nitrogen gas then Curtius rearrangement of the resulting nitrene to afford isocyanate intermediate (non-isolable) followed by exo-trig ring closure (Scheme 3). The structure of 4 was substantiated from its IR spectrum which lacked ν N=C=O of isocyanate group and only displayed the stretching absorption bands of oxazinone CO and NH at ν 1732 & 3175 cm -1 , respectively. The 1 H-NMR spectrum was a real evidence for its existence as a mixture of lactamlactim tautomers through which two signals at δ 13.28 and 13.25 ppm displayed, both integrated to one exchangeable proton of NH-CO N=C-OH grouping. Moreover, the mass spectrum exhibited the correct molecular ion peak at m/z 253 (46%) as well as the base peak at m/z 252 (100%) representing [M-1] cation. The same product 4 was obtained via pyrolysis of the azide by fusion in neat.
When azide 3 was allowed to react with benzylamine in dry benzene at room temperature, the benzylurea derivative 5 was obtained, while the reaction in refluxing benzene for 1 h resulted in the formation of pyrimidinone derivative 6. The structures of 5 & 6 were deduced from their analytical and spectral data. The IR spectrum of 5 revealed a broad band at 3247 cm -1 attributable to NH groups, as well as, the characteristic absorption bands for C=O of ketone and amide groups at 1671 & 1643 cm -1 , respectively. The 1 H-NMR spectrum of 5 showed that CH2CO protons were magnetically non-equivalent which may be attributed to hydrogen bonding between PhC=O and NH (Cf. Experimental). The 1 H-NMR spectrum of 6 was devoid from the splitting pattern of CH2CO group and showed a singlet band at δ 4.8 ppm due to benzyl-CH2 protons, as well as, two signals at δ 10.03 and 10.04 ppm, both integrated to one exchangeable proton of NH-CO N=C-OH grouping. The reaction of 3 with hydrazine was also dependent on the reaction conditions. Thus, stirring 3 with hydrazine in benzene at room temperature afforded the semicarbazide derivative 7 while at refluxing conditions; 1-aminopyrimidinone derivative 8 was produced. The IR spectrum of 7 showed a broad band at 3331 & 3208 cm -1 attributable to NH & NH2 groups, as well as, the characteristic absorption bands for C=O of ketone and hydrazide groups at 1700 & 1659 cm -1 . The 1 H-NMR spectrum of 7 exhibited geminal coupling of two magnetically non-equivalent protons of CH2 (J=16.4 Hz) as evidence for the aforementioned hydrogen bonding (Cf. Experimental).

Scheme 2. Reactions of azide 3
The aromatic amine, p-toluidine, failed to affect decomposition of the azide 3 at room temperature, a behavior that may be attributed to the weak nucleophilicity of the p-toluidine nitrogen atom. But, when the reaction was carried out in dry benzene at 90 o C for 2 h, Base-catalyzed decomposition occurred with the formation of the ptolylurea derivatives 9. On the other hand, Carrying out the reaction under reflux conditions for 6 h led to the construction of p-tolylpyrimidinone derivatives 10. The IR spectrum of 9 provided the characteristic absorption bands for two NH & two C=O of ketone and amide groups at 3317 & 3209 cm -1 and 1699 & 1650 cm -1 . Also, the geminal coupling between CH2 protons (J=16.0 Hz) was clearly shown in its 1 H-NMR spectrum indicating to hydrogen bonding (Cf. Experimental). The formation of the oxazinone and pyrimidinone derivatives may be represented by

Introduction
The genus Arum (Araceae) is native to Asia, Europe, and northern Africa. The number subspecies of this genus is not definite: while some researchers consider 29 subspecies, 1 the "U.S. National Plant Germplasm System" counts 44. 2 However, the number of subspecies that have known (reported) traditional uses and were reported in current studies for biological/medicinal activities hardly exceeds two dozens. Archeological evidence indicate uses of Arum by humans since ancient times. 3 Arum subspecies are well known for their thermogenesis. 4 This is to say that alteration of light and dark in the environment of the plant stimulates the primordia of the male plant to produce salicylic acid that triggers thermogenic reactions. For some subspecies like A. italicum and A. maculatum, the temperature of the flower can be higher by 15-25 C than the surrounding air. This phenomenon is one of two major pollination strategies that aim to attract potential pollinators like insects. The other strategy is releasing a very strong odor that attracts insects. In most subspecies of Arum, this odor is foul (dung, A. palaestinum, A. dioscoridis, A. elongated and others) but in some subspecies, it can be from not perceptible (A. jacquemontii) to even pleasant (A. gratum). 4 In addition to many volatile amines that will be presented in next sections, many compound families are represented in these pollination odors. 4,5 Some important compounds are shown in Figure 1.
Detailed study of the floral odor of A. italicum was published earlier in 2004, where several methods of isolation and trapping volatile compounds were used. 6 In this study, very similar results were obtained comparing with the previously cited publications (4,5), and only stereochemical and structural isomerization can be noticed, comparing with the compounds shown in Figure 1. Most texts this property is mentioned and potential users are explicitly warned. Modern research approved this as shown in below (discussion).
b) It is notable that the most important traditional uses of Arum are for nutritional purposes then by medicinal use as an anticancer agent. This using field is also in agreement with current research results.

Modern research reports of Arum subspecies
Many Arum subspecies were studied so far, where the most investigated are A. dioscorides, A. maculatum, and A. palaestinum. It is interesting to see that unlike other plant families that are used by humans for millenia, modern research of Arum started just little more than three decades ago, while other plant families are being studied for much longer periods of time. Many medicinal and other biological activities of Arum plants were reported. In Table 2 a summary of these reports is presented.

Discussion
Arum subspecies are known and used by humans since ancient times. But new subspecies are still identified once in a while. Arum megobrebi was identified and classified as wild subspecies of Arum that grows in Turkey and Georgia. 107 Reading data in Tables 1 and 2  It is interesting to pay attention to A. cyrenaicum, an endemic subspecies that grows wild only in Libya. In reference 8, authors report two traditional uses of this plant (food and ornaments), and it is interesting to notice that the used parts are corms, not leaves, contrary to most Arum subspecies, where corms are highly toxic. But these authors have mistakenly classified this plant into the Poaceae family, while the correct classification is in the Araceae family. 55 One of the most important properties of Arum that was consistently mentioned by traditional users and approved by modern research is the toxicity of these plants. Despite being recommended for use as food and medicine, the toxicity of Arum is indicated in most texts. 108 Modern reports rank Arum subspecies as one of the most important causes of children poisoning in Brazil. 109 Among these, A. italicum is responsible for the largest number of poisoning cases, and all parts of the plant are toxic.
Toxicity of Arum subspecies results from several single compounds or compound families. Calcium oxalate is one of the primary toxic compounds in Arum plants, 55 but it decomposes with cooking. The same occurs to cyano glycosides such as triglochinin, 76 a toxic compound present in Arum, that its structure is shown in  Among reported Arum subspecies in the toxic context, A. maculatum is the most published by current research publications so far. One of the earliest reports was published in 1861, and it presents some poisoning cases. 110 Part of this toxicity is due to the presence of toxic odorants, especially volatile amines. 111 In addition to oxalates and cyano compounds, the toxicity of A. maculatum is intensified by alkaloids and saponins. 112 The orange-colored fruits of A. maculatum are very attractive yet very poisonous, and they are responsible for most poisoning events caused by this plant. 113 The toxicity of A. palaestinum is also known and published: ethanolic extract of the plant was found toxic to the liver of female rats. 114 Despite that, unlike other natural, plant-derived anticancer therapies, A. palaestinum has no herb-drug contradictions with synthetic drugs. 115 An interesting modern research report presented in Table  2 is about A. Conophalloides. 54 All 18 compounds identified in the essential oil of this plant do not contain nitrogen. No amines or alkaloids. This situation can be understood from two reasons: nitrogen containing compounds, especially amines are volatile and alkaloids are not volatile and mostly water soluble, so they are not present in the essential oil that contains mainly hydrophobic compounds.    76 olated and identified The toxic cyanogycoside was is Fatty acids contents Fatty acids of seed oil were isolated by picolinyl esterification and purification. In addition to medium chain acids (C14:0), acids with 77 aromatic residues (including pyridyl) were detected Pro-inflammatory A monocot lectin (ptotein) was isolated from the tubers of the plant. 78 inflammatory activity -It acts as agglutinin and has pro Insecticidal Mannose binding lectin was isolated from the tubers. It binds to the 79 glycosylated insect gut receptors Cytogenetic 80 Aqueous extract inhibited cell mitosis of bone marrow of mice Antioxidant capacity of food Antioxidant capacity of leaves were tested in three forms: fresh, 81 powder and stored. All forms showed similar capacities Analgesic Aqueous extract analgesic activity was compared with that of declofenac-Na and morphine. It was more active than the first and 82 had similar activity of the second Antibacterial 83 Ethanolic extract tested against 7 typyes of bacteria: weak Antibacterial Four extracts (petroleum ether, chloroform, ethyl acetate and 70% methanol) of aerial parts were tested against two bacteria. 84 Hydromethanolic extract showed highets activity Antioxidant Methanolic extract of whole plant was by DPPH assay and found 85 re than ascorbic acid very active, even mo Essential oil, antibacterial, antioxidant Essential oil was tested for antibacterial (

A. palaestinum
New Alkaloid (S)-3,4,5-trihydroxy-1H-pyrrol-2(5H)-one (2 in Figure 6) was isolated from the aqueous extract and characterized. The ethyl acetate extract showed strond antioxidant and sufficient anticancer 90 activities Antioxidant, antidiabetic Aqueous and methanolic extracts were prepared and tested for antioxidant activity (DPPH): moferate. This result is in agreement 91 with total phenolics content and antidiabetic traditional use Anticancer modern herbal medicine Aerial parts, especially leaces are used as anticancer agents in modern herbal Palestinian medicine. It is used raw, cooked (food) or 92 as a decoction Anticancer, antioxidant Aqueous and ethanolic extracts were tested for anticancer and antioxidant activities. Anticancer was very strong (aqueous >> 93 ethanolic), antioxidant was weak (aqueous > ethanolic) Diketopiperazines Two new alkaloids were isolated from the aqueous extract and 94 characterized. Only 3 in Figure 6 showed cytotoxic activity Antimicrobial 70% Aqueous ethanol extract was tested for antibacterial activity against six types of bacteria (weak), and for antidermatophyte 95 activity (2 fungi Two of the major (>5%) compounds are both chemically and biologically interesting. These are the structurally isomeric alcohols T-cadinol (8.9% in the essential oil of A. Conophalloides) and T-muurolol (24.4%). Their structures are shown in Figure 3.

Figure 3. Structures of T-cadinol and T-muurolol
These compounds have many biological activities such as antibacterial of T-cadinol. 116 They are present in relatively high concentrations, and it might be useful to try to isolate them from other subspecies of Arum. A. italicum produces a wide variety of amines during the flowering season. 56 Some of these amines are very interesting regarding the number of nitrogen atoms that they contain. In Figure 4, the structures of three of these amines are shown (with metformin).
The structures of agmatine and metformin are relatively close. Metformin is very well known synthetic antidiabetic drug, and the great medicinal potential and activities of agmatine are being studied, including antidiabetic activity, 117 , but this research must be expanded.
Antibacterial and antimicrobial activities are tested for almost every studied medicinal plant. In the case of Arum subspecies, some were reported, and these reports are not consistent. Even after taking into account the different subspecies, various parts of the plants that were extracted and the various solvents that were used, the overall reporting is confusing and even contradicting. 60,61,68,69,83,84,95 For example, M. Obeidat et al. 60 reported that they tested four extracts (water, ethanol, methanol and acetone) and found the aqueous extract most active. On the contrary, A. Ucar Turker and her colleagues, 68 used aqueous, ethanolic and methanolic extracts, and the aqueous extract was inactive.
In 1994, M. Della Greca et al. isolated and characterized phytosterols and hydroperoxy sterols from A. italicum, where some were new. 70 Despite the fact that similar compounds were isolated from other plants and marine animals, 118 and proved to have significant biological activities, a follow-up study was never reported. It is worth trying to find this compound family in other Arum subspecies, characterize them and test them for biological activities, especially antimicrobial and antifungal activities. The presence of the peroxy group ensures oxidant activity, while the entire compound is hydrophobic and can penetrate the lipophilic membranes of microbes and fungi. The structures of the new hydroperoxy sterols that were reported by M. Della Greca et al. are presented in Figure 5. Alkaloids are the major compound family in Arum subspecies. Their toxic and psychoactive influence affected users of this genus since very ancient times. But the isolation and characterization of these compounds from Arum started relatively late. 89 The polyhydroxy alkaloid that was isolated in the same year, 90 provides an interesting starting material for synthetic purposes. Two other new alkaloids were reported later, and they have even simpler structures. Arum subspecies can be the natural source for such heterocyclic alkaloids. See Figure 6.  99 reveals some confusion regarding the presence of caffeic acid in A. palaestinum. The first reports that it is present, while the second clearly indicated (ND) that it is not. But studying other reports show that this compound is present in A. palaestinum, and many of its derivatives. 90,100 In the same sense, it is not clear why M. M. Farid et al. 102 claim that isovitexin was "isolated (by them) for the first time from the studied taxa," while they reported the isolation of the same compound in one of their earlier works. 96 Conclusions 1) Many subspecies of the genus Arum were never studied, which is a very vast field of future potential research.
2) It is important to invest more research in nitrogen containing compounds of Arum subspecies. The structures of the known compounds so far (very few ssp.) indicate a high potential for antidiabetic activity, which might result from a single compound or synergy of several compounds.
3) Antibacterial activities of Arum subspecies need further studies and organization. 4) Some activities like the antidiabetic potential of Arum were hardly investigated. There is an urgent need to expand the research of these activities.
5) Very few attempts were made so far to prepare synthetic modifications of active natural products isolated from Arum which could be intensified.

Introduction
Emil Knoevenagel (in 1890) developed a method for the synthesis of substituted alkenes, by the condensation of an aldehyde with active methylene compounds in the presence of base and water. Knoevenagel condensation is typical C-C bond forming reaction in organic synthesis. This reaction is useful to generate a variety of intermediates which are used in the synthesis of pharmaceutical precursors; because of this, Knoevenagel condensation has been extensively studied by researchers. Scientist and academicians are still inventing novel methods and catalysts for Knoevenagel condensation. Several methods have been developed by using the microwave, 1 or ultrasonication, 2 photochemical condensations with fruit extract as a catalyst, 3 solvent free conditions. 4 Recently Franca Bigi et al., 5 reviewed Knoevenagel reactions in an aqueous medium with and without a catalyst. According to Franca although reaction involves a dehydration step, the reaction can be carried out in water. Following this interpretation, we have carried out the Knoevenagel reaction of pyrazole aldehyde in water but resulted in lower yield due to less solubility of pyrazole aldehyde. Then we have carried out the reaction in the water-ethanol mixture and obtained a high yield of products; these results prompted us to investigate this reaction further.

Experimental
All chemicals used were of the synthetic grade. The solvents were distilled before use. The progress of the reaction was monitored by TLC using ethyl acetate: nhexane system. Melting points were recorded by using the open capillary method and are uncorrected. The Ultrasonicator used was made by Cyberlab Ultrasonic Stericleaner model number CB2080 with operation voltage 220 V AC and electric cycle 50/60 Hz. IR spectra were recorded on Shimadzu IR Affinity 1 instrument using KBr discs. H 1 NMR was recorded on BRUKER Avance II 400 NMR Spectrometer using DMSO d6 as a solvent. The mass was recorded on WATERS, Q-TOF Micro mass (ESI-MS) using methanol as a solvent.

General procedure for the Knoevenagel condensation
In 50 mL round bottom flask pyrazole aldehyde (1 mmol), malononitrile (1 mmol), were taken in 10 ml water-ethanol (1:1) mixture and stirred for 3-5 minutes to mix the reaction mixture; after that ammonium carbonate (20 mol %) was added. The resulting reaction mixture was stirred for 3-20 minutes at reflux temperature, and the reaction was monitored by TLC. After the completion of the reaction, the reaction mixture was allowed to cool down to room temperature and then filtered off, washed with water and dried. Similarly, other derivatives were also prepared ( Table-2). Similar results were obtained when the reactions were carried out using sonication method (Table 2).

Result and Discussions
Owing to the importance of Knoevenagel condensation reaction to synthesise pharmaceutical intermediates, we have developed a green and efficient method, for the synthesis of heterocyclic, substituted alkenes, by reacting pyrazole aldehyde, malononitrile in the presence of ammonium carbonate (20 mol%) in water: ethanol (1:1) mixture at reflux temperature.
To optimize reaction conditions we have performed the reaction of 1,3-diphenyl-1H-pyrazole-4-carbaldehyde with malononitrile and ammonium carbonate which was considered as a standard model reaction. In search of a suitable solvent, we have achieved reaction by using different solvent mixtures and results were summarized in (Table1 ). Table 1. The reaction of 1,3-diphenyl-1H-pyrazole-4-carbaldehyde and malononitrile in the presence of 20 % and ammonium carbonate in 10 ml of the solvent system at reflux temperature.
Water: ethanol (1:1) mixture is proved to be the best solvent system for this reaction. If the reaction was carried out either in water or ethanol; yield of product was decreased due to the higher solubility of either pyrazole aldehyde or ammonium carbonate. The amount of ammonium carbonate has a large influence on the reaction time and yield, the reaction in the absence or presence of 10 % ammonium carbonate only 30 or 60 % yield could be achieved, respectively, and unreacted aldehyde remained back even after 1 h reaction time.  We have also carried out the same reaction by using sonication to increase yield and reduce the period of reaction. But much difference was not found (Table 2).

Conclusion
The reported method described a simple and fast way for the condensation of pyrazole aldehydes with malononitriles, in 1:1 EtOH: an H2O solvent mixture in the presence of 20 mol % (NH4)2CO3 and under 10 min reflux or sonication.

INTRODUCTION
Making stable thin sections of biological tissues with their subsequent staining and analysis using light microscope helps to analyze animalcular texture of the organs. 1 -9 Recently paraffin, celloidin and gelatin pouring of tissues allow to extract histologic specimen applying different ways of staining and to carry out immunohistochemical investigations. At the same time, the quality of tissue structures' preservation remains moderate. 9 The best preservation of tissue structures is being reached at tissues' epoxy injection followed by preparation of semithin sections. Epoxy resins provide optimal tissue morphology at both the light and the electron microscopic level and therefore enable correlative studies on semithin and thin sections from the same tissue block. 6 In the three-dimensional reconstruction of a neuronal structure, it is imperative that ribbons of semithin or ultrathin sections be obtained. Resin-embedded semithin sections display better structural details than paraffin-embedded sections.
The aim of the current research is to show potential for light optical examination of blood vessels of tissues included in epoxide resins for a long-term. Small pieces of tissue have immediately put in a cold mix of paraformaldehyde in a sodium cacodylate buffer and glutaraldehyde for 12 hours at 4 °C with following post fixation in 1% OsO4 solution for 2, then dehydration in ascending series of spirits; saturation in a mixture of acetone and Epon resins of different proportions to make gelatinous capsules were performed.

Reagents
Observation under a light microscope: Semifine (semithin) epoxy sections with 1 m thickness were made using LKB (Swedish) and Reichert (Austria) tools and the obtained semithin epoxy sections stained with Azur 2 and studied under a light microscope supplied with 40 x10 ocular lens.

RESULTS
The research of the biopsy material (right auricle atrial of myocardium) and the experimental material (liver) had been carried out. The material had been collected in different years. So biopsy materials of human myocardium were received in 1997 during mitral valve replacement in patients with rheumatic heart disease and coronary artery disease.
Biopsy material of liver was received in 2010 during the investigation of chronic stress. Sampled material was poured into epoxide resins by standard method for transmission electron microscopy. It must be mentioned that for the last years the collected material had been stored at different temperature conditions as low so high.  The investigation of those materials poured into epoxide resins in different years was carried out. It was necessary to check up how our invention worked 7 on material collected years ago. Nowadays the research on semithin epoxide sections of biological material embedded in epoxide resins provokes more and more interest as it allows seeing thinner morphologic picture under a light microscope. Azur II used by us with sodium borate shows quite interesting results in carrying out current research in diagnosing work on blood vessels in biopsy as it can be seen in Figure 1, 2 and 3.
That pointed out that the current method of investigation allows diagnosing micro circulatory changes as in pathology of a human being so during experimental researches irrespective of the years the material had been collected and the temperature conditions kept at its storing.

DISCUSSION
TEM-examination of ultrathin sections is usually a prerequisite for the researchers working with semi thin epoxide sections of epoxy injected tissue samples.
Semifine epoxide sections are stained in a standard way by Toluidine blue, Methylene blue, AZURE II in different combination. It should be noted that AZURE II almost is not used separately for staining. 2,3 The most common stain used in the electron microscopy lab for thick sections is toluidine blue. 1,4,8,10,11 Unfortunately its general lack of polycromasia makes it unsuitable for photomicrography. Much better results are obtained for general work as well as for photomicrography (especially with the use of filters for the black and white film) with the methylene blue, azure II combinations. 5 They are prepared in advance and most EM-Labs only stained by Toluidine blue because only an approximate morphological information on the area to be sectioned for ultrathin is sufficient.
Toluidine blue is indeed commonly used for stain semithin sections to show the general structure of the cell. Methylene blue-azure is an alternative. 7,9,11,12 In our research, we used only AZURE II 1% solution prepared on 1% sodium borate by our staining method which turned out to be quite applicable for analyzing blood vessels. Our results show to the well preservation of tissue in epoxide resins for its studying under light microscope irrespective of the time factor and temperature conditions' storage.

CONCLUSION
Collected in different years biological material with shelf life from 18 to 5 years was useful for purposes of light optical research of microvessels on semithin epoxide sections.

INTRODUCTION
Pycnanthus angolensis (Welw.) Warb. syn. P. kombo (known as African or wild or false nutmeg) was originally native to the forest zones of West and Central Africa but now cultivated in and around the world. 1 Different preparations of the plant are employed in diverse African folklores to treat chest infections, diabetes, lumbago, wounds, arthritis, anaemia, mouth-thrush, malaria, leprosy, toothache, infertility, sexually transmitted disease and skinfungal infections. [2][3][4] The larvicidal and antitumor potentials of the plant have been studied 5-6 while reports of isolation of flavonoids and terpenes from the bark and roots and quinones from the leaves abound. 7 This present investigation aimed at isolating the compound(s) from the ethyl acetate fraction which demonstrated the highest antimicrobial activity in a previous bioactivity-guided fractionation study of the plant. 8 In addition, the compounds so obtained will be screened for antimicrobial activity with the aim of confirming or disproving the claims highlighted in traditional medicine especially for the treatment/management of bacterial infections.

MATERIALS AND METHODS
The fresh leaves of P. angolensis were collected around April, 2016 within the precinct of University of Uyo, Akwa Ibom State, Nigeria. The plant had previously been identified in a study. 8 Immediately after collection, the plant was dried in a laboratory oven (Gallenkamp, England) at 40 0 C for 48 h and the resultant material powdered on an electric mill (Uniscope, England).

Extraction and isolation
The dried powder (1.1 kg) was exhaustively extracted with 50 % EtOH (3 x 5L) at room temperature (27± 2 0 C) for 72 h. The resultant crude extract mixture was filtered, concentrated in vacuo on a rotary evaporator. 250 g of dried crude extract was obtained and then stored in a desiccator prior to further use. Consequently, 15 g of the extract was partitioned using H2O: EtOAc (8 x 200 mL). The combined ethyl acetate fractions were evaporated to dryness to give a brown solid residue. Hence, 1.

Antimicrobial tests
The microorganisms used in this study, namely; Staphylococcus aureus (ATCC 21824), Escherichia coli (ATCC 2353) and Candida albicans (NCYC 106) were clinically isolated from specimens of diarrheal stool, abscesses, necrotizing fasciitis, urine and wounds obtained from the Medical Laboratory, University of Uyo Health Centre, Uyo. The clinical isolates were collected in sterile bottles, identified and typed by convectional biochemical tests. 9-10 These clinical microbes were then refrigerated at -5 0 C. The agar plates used were prepared by adhering to the manufacturer's instructions. The media and plates were sterilized in an autoclave at 121°C for 15 min.
The hole-in-plate agar diffusion method was used observing standard procedures for Nutrient Agar-CM003, Mueller-Hinton-CM037 (Biotech Limited, Ipswich, England) and Sabouraud Dextrose Agar (Biomark, India) in respect of bacteria and fungus respectively. The inoculum of each micro-organism was introduced into each petri-dish (Pyrex, England). Cylindrical plugs were removed from the agar plates using a sterile cork borer (Simax, India) to produce wells with a diameter of approximately 5 millimetres. The wells were equidistant from each other and the edge of the plate. [11][12] Concentrations of 20 mg mL -1 of crude extract, 10 mg mL -1 of ethyl acetate fraction, 2 mg mL -1 of NG-2 and NG-4c were introduced into the wells. Also, different concentrations of 10 g mL -1 Streptomycin (Orange Drugs, Nigeria), 1 mg mL -1 of nystatin (Gemini Drugs, Nigeria) and deionized water were introduced into separate wells as positive and negative controls respectively. 5,[13][14][15] The experiments were carried out in triplicates. The plates were labelled on the underside and left at room temperature for 2 h to allow for diffusion. The plates were then incubated at 37 ± 2 0 C for 24 to 48 h. Zones of inhibition were measured in millimetres (mm) with the aid of a ruler.

Structural elucidation
The mass spectra of the compounds were obtained on Kratos MS 80 (Germany) while the infra-red analyses were done on Shimadzu FTIR 8400S (Japan).

Antimicrobial screening
The spectrum of microbes employed in the sensitivity tests was narrow, encompassing one each of gram positive (S. aureus) and gram negative (E. coli) bacterial strains and a fungus (C. albicans). The results displayed in Table 1 show that the crude extract, ethyl acetate fraction, NG-2, and NG-4c were inactive against S. aureus and C. albicans. However, the two compounds were remarkably bacteriostatic against E. coli. This result was unexpected because gram-negative bacteria are well known for their unique resistance to antimicrobial agents. This resistance is believed to be due to the nature of the cell envelope of these organisms which unlike gram-positive organisms possess a sophisticated three-layered envelope which does not allow permeation of external agents. Also, both compounds demonstrated no antifungal activity against C. albicans. This particular observation was to be expected because of fungal strains especially Candida spp. limit the permeation of substances because of their integral structures which are pleomorphic and facultative in nature hence, resembling those of higher plants. 16 It is instructive to mention that derivatization studies are currently on-going in our laboratories with the aim of improving on the observed activity.

CONCLUSION
The isolation of the two compounds is being reported for the first time from the ethyl acetate fraction of the plant. Hence, ethyl linalool and diethyl phthalate are expected to serve as chemotaxonomic markers for this species and the genus, Pycnanthus in general. Furthermore, the results of the antimicrobial sensitivity tests lend some credence to the use of this plant especially in the treatment or management of the bacterial disease.

Introduction
The world around us is in a constant motion. Functionally interconnected main types of mechanical motion (translational, rotational and oscillatory) determine the dynamic stability of systems. Vast theoretical and experimental experience in physical and mathematical properties of simple and complex compounds and principles of their self-organization at different scale levels of such conformation has been gained till now. But the problem of establishing the most general regularities of these processes is topical. "However the science is still far from making it happen in the general form". 1 Thus, applying the entire set of analytical and qualitative methods, the celestial mechanics provides the solution for many problems on the motion of solids, for example. 2,3 But some other issues of celestial mechanics require further discussion, for instance, the functional dependence of rotational and orbital motion of planets, as well as the initial principles of forming the direct and reverse motion of planets. Therefore in this paper we attempt to investigate such problematic issues with the help of conception of corpuscular-wave dualism proposed earlier. 4 Initial criteria 4 1. In the systems in which the interaction proceeds along the potential gradient (positive work), the resultant potential energy is found based on the principle of adding reciprocals of corresponding energies of subsystems. Similarly, the reduced mass for the relative motion of the isolated systems of two particles is calculated.
2. In the systems in which the interactions proceed against the potential gradient (negative performance) the algebraic addition of their masses, as well as the corresponding energies of subsystems is performed (similar to Hamiltonian).
3. Two principles of adding energy characteristics of structural interactions can be transformed onto the corpuscular-wave dualism processes.
Corpuscular processes flow in all interactions along the potential gradient, and wave dualism corresponds to the interactions against the potential gradient.
4. Act of quantum action expressed via Plank's constant is narrowed to the energy equilibrium-exchange redistribution between the corpuscular and wave processes. Here

Pe = wr,
where wenergy of a free electron, rits classic radius.
6. It is assumed that during the rotational-translation motion of an electron, the energies are redistributed in the system "particle-wave" that is demonstrated via the angular vector of such motion (winding angle) -ϴ.

  
This angular vector of electron motion is quantized by an integer number through the tangent square of this angle: tg 2 φг = 2; tg 2 60º = 3; tg 2 45º = 1, where φг = 54.73 0a socalled "geodesic angle", which is widely spread in engineering, for example, in spaceship production.
The quantum functions of square tangent k = 1, 2, 3 numerically determine the ratios of two triangle legs, whose values characterize energy dependencies via axial and circumferential stresses in the system with corpuscular and wave processes. 7. In quantum mechanics the ratio between the particle magnetic moment and its mechanical moment is the magnetomechanic ratiog. At the same time, gs = 2, if the electron magnetic moment is conditioned only by the spin component and g = 1, if it is produced by the electron orbital motion. Their ratio gs/g = 2 that, the same way as tg 2 φг = 2, characterizes the corresponding corpuscular-wave dependencies in this approach.

Equation of dependence of rotational and orbital motion of planets
The foregoing principles of corpuscular-wave mechanism give the possibility to consider from the unified positions many structural and dynamic processes, different in nature and scale. For example, the characteristic of spin-orbital interactionfine structure constant =r/, where relectron classic radius, λits Compton wavelength.
Formally, but similarly: interaction force of two long conductors with the current proportionally to the ratio l/2r, where llength of conductors, rdistance between them.
And the number 2π widely used in physical regularities, equals the circumference ratio to its radius: 2π = l/R.
In these examples, as in many other, this approach allows evaluating structural interactions based on the ratios of corpuscular and wave spatial-energy parameters in each action. Obviously, such principles are also demonstrated in Kepler's third law, which can be given as follows: 3 (1) where: Ggravitational constant, mplanet mass, adistance to barycenter (system mass center), Tplanet revolution period.
Since the masses of planets are rather small in comparison with their distance to the sun, then at the first approximation they can be considered as mathematical points and the equation of mathematical pendulum period can be applied to them: where gfree fall acceleration.
The average radius of the planet orbital motion -R can be taken as the pendulum lengthl. Then, similarly to the foregoing dependencies, we can introduce the planet radius r into the numerator of formula (2) and then we have: (4) This expression in the units m/s 2/3 satisfies the principle of corpuscular-wave mechanism for space macrostructures. But in Kepler's third law only the orbital motion was considered, but in this casetwo motions, each of which has its own wave part. Therefore, the interference of coherent waves occurs.
Similar to the foregoing examples, the coherence can be considered as the ratio of the difference of the travel path to the length of the coherent wave . The interference principle is most easily performed for liquid-gaseous planets (planets in Jupiter system) as shown in Table 1: -boosting of oscillations during the direct rotation of planets.
-damping of oscillations during the reverse rotation of planets. Here ninteger number.
The intensity of wave propagation depends on the medium density and its distribution in the planet volume.
The value characterizing the planet density increase towards its center is called "dimensionless moment of inertia"(I * ). The ratio of average values of I * for solid and liquid-gaseous planets based on different data 3,5,6 is within 1.4 -1.45 as demonstrated in Table 2.  Such property for solid planets is taken into account in Table 1 and equations (5, 6) by introducing the values n/2 and (2n+1) 1/2 . Such approach also refers to Mercury as it is the nearest to the sun, to its liquid-gaseous structure.
In general, the application of corpuscular-wave mechanism to space macrosystems explains the specifics of formation of direct or reverse rotation of the planets.

Rotation angles of planets
In physical sense, the parameter β characterizes the motion difference of interfering waves, and γwavelength. The average value of β = 732.5 m/s 2/3 with the deviation of most of the planets under 2 % (apart from Uranus).
The equation tg 2 ϴ=k was used to evaluate quantum transitions in atoms. 4,7 The squares and cubes of initial parameters are applied in Kepler's equation and other regularities of space macrosystems. In this approach, as the calculations demonstrated, the semi-empirical equation is performed: where ϴrotation angle of planets.
For Earth β = 755.2 and based on equation (7) ϴ0=66.455°. For more accurate calculation, taking into account some analogy of macro-and microprocesses, we use, as before, the experimental quantum correction in the form of a0 = 1.0023293 following the equation: The calculation by equations for Earth (7,8) gives the value 1=66.560º with the deviation from the experimental value by 0.007 %. The sun has the same value of the rotation angle.
As applicable to the rest of the planets, the value  (where β0 -β value for Earth) is introduced in equation (8) based on the following equations: or (9,9a) or (10,10a) The formulas (9 and 9а) are performed for the planets, whose rotation angle is less than Earth's one. For the other planets the formulas (10 and 10а) are performed. Those are the planets, which are in the beginning of the planet subsystem by the value of the dimensionless moment of inertia (Mercury and Jupiter), as well as the planets with the reverse rotation (Venus and Uranus). The calculation results are given in Table 1.
The coefficient 4/3 has been applied before for the comparative evaluation of quantum transitions with different complexity types. 4 In this research the average ratio of the angles by the experimental data 3,5,6 given in Table 3 also has the value 1.336 ≈4/3.  For the value  the influence of the medium density distribution is taken into account via the transition factor from one distribution level to another. Since for Jupiter in Table 2 the value I * is less in comparison with Earth's I * in 1.45 times, therefore, in the calculations  = 0.9478 1/2 . On the contrary with Jupiter, for Neptune I * increases in 1.45 times, therefore, in the calculations  = 0.96793 2 . All the calculation results are in good according with the experimental data.

Conclusions
1. Semi-empirical equations of the dependence of rotational and orbital motion of planets are obtained.

Introduction
Coumarin derivatives are an important class of compounds and are well-known for their biological activity. Many of heterocyclic derivatives on the basis 2H[1]benzopyran-2-one play an important role in various life processes, and they are found as an ingredient in the plant world. Many such derivatives exhibit various biological activities, 1 such as anticoagulant, 2 antimicrobial, 3 antibacterial, 4, 5 antifungal 6 and antimalarial. 7 Some of coumarin analogs also exhibited antioxidant, 8,9 antitubercular 10 and anticonvulsant activity. 11 It was reported that a significant number of substituted derivatives of benzopyran-2-one also show, sedative, 12 analgesic and antiinflammatory, 13 anti-HIV, 14 and hepatoprotective activity. 15 According to that, many of them have found widespread usage in pharmacies. 16 On the other hand, azetidin-2-one derivatives reported having a wide range of biological activities, 17 including those antimicrobial, 18 anti-fungal, 19 anti-convulsant, 20 and anti-HIV activity. 21 The biological activity of these derivatives is conditioned by their structure, so the presence of different substituents on the benzopyrone ring indicates their impact on the type and potency of biological activity. Despite continuous efforts, the relationship between structure and biological activity of these derivatives so far has not yet been sufficiently clarified. Extraordinary biologically importance of such derivatives on the basis of thiazolidine-4-one has generated a constant interest for their synthesis and research. In continuation of our previous studies and attempt to synthesize the new derivates, 22,23 in this paper we had intended to synthesize some new amino-phenylamino)-3-nitro-chromen-2-ones and substituted thiazolidyne-4-ones with benzopyran-2-one moiety, which could serve as pro-pharmaceutical products.

Methods and materials
The compounds are synthesized using commercial reagents of Aldrich company as precursors under catalytic conditions. Reactions are monitored by TLC using Merck Kieselgel-60 (F-254) as the stationary phase and a mixture of benzene, toluene, glacial acetic acid (v/v/v, 80:10:10) as the mobile phase. The synthesized products are purified by crystallization from methanol and ethanol. Melting points are determined used a paraffin oil bath with an open capillary tube. IR spectra are recorded in KBr discs on Shimadzu 8400xFT-IR spectrometer with 4 cm -1 resolution. 1 H-NMR and 13 C-NMR spectra are recorded in DMSO on UNITYplus-500"NMR 1" spectrometer. Chemical shifts were reported in ppm downfield from TMS as internal standard (δ 0.00). Examination of the antibacterial activity of the synthesized compounds is done using standard discs (d=5.0 mm, maximum capacity 10 pg) on the basis of Standard Disc Method. Standard discs are previously impregnated with 2 mg mL -1 , 4 mg mL -1 and 6 mg mL -1 solutions of compounds in -DMF.

General procedure for preparation of 4-[4-(benzylideneamino)-phenylamino]-3-nitrobenzopyran-2-ones, 4(a-c)
Product 3 (0.3 g, 1.0 mmol) is dissolved in 20 mL of absolute ethanol and to this mixture was added in small portions 1.5 mmol of aromatic aldehyde (benzaldehyde, salicylaldehyde or 3-nitrobenzaldehyde respectively) dissolved in 10 mL of absolute ethanol. Then there was added 2 drops of piperidine as a catalyst and the mixture was mixed for 15 min at room temperature and refluxed for 10 to 12 hours. After cooling, the mixture is concentrated and the crystals are filtered off under reduced pressure, then washed with 2x1 mL of ethanol and dried in the air. crystallization of the products 4a-4c was conducted from ethanol or methanol.
In the IR spectrum of 4a, an absorption signal appeared at 3461.45 cm -1 which are responsible for ν(NH) stretching vibrations, while absorption signal at 3075.24 cm -1 corresponds to ν(CH) of the aromatic ring. At 1694.15 cm -1 appeared the absorption signal which response to ν(C=O) stretching vibrations, whereas the sharp peak at 1642.76 cm -1 and a signal at 1603.07 cm -1 correspond due to ν(C=N) and ν(C=C) stretching vibrations of the aromatic system. The characteristic absorption at 1551. 15  Absorption at 1221.34 cm -1 is also characteristic for stretching(C-O-C) vibrations of the lactonic ring, whereas the sharp peak at 756.85 cm -1 is characteristic for bending δ(C-H) oop vibrations of the aromatic ring. On the other hand, signals from 1 H-NMR spectrum correspond to the absorption of respective protons. A proton singlet displayed at 8.35 ppm resulting from N=C-H. Also in the 13 C-NMR spectrum is displayed a signal at 165.4 ppm which corresponds to the C=N carbon.
The IR spectra of compound 4b, showed the absorption signal at 3451.94 cm -1 which is responsible for ν(NH) stretching vibrations, the broad band at 3307.35 cm -1 correspond to stretching ν(OH) absorption, and a signal at 3079.24 cm -1 resulted due to ν(CH) stretching vibrations of the aromatic ring. The sharp peak at 1684.15 cm -1 resulted from ν(C=O) stretching vibrations, whereas signals at 1608.97 and 1552.51 cm -1 result from ν(C=N) and ν(C=C) stretching vibrations. The peak at 1514.34 cm -1 corresponds to absorptions of asymmetric ν(NO2) stretching vibrations, while the one at 1320.49 cm -1 due to symetric ν(NO2) stretching vibrations. Absorption signal at 1211.64 cm -1 is characteristic for stretching vibrations of the lactonic (C-O-C) system, and the sharp peak at 761.67 cm -1 resulted from δ(C-H) bending oop vibrations of the aromatic ring. In the 1 H-NMR spectrum, besides multiplets of aromatic protons, a proton singlet resulting from N=C-H appears at 8.38 ppm. In the 13 C-NMR spectrum are also displayed a signal at 163.8 ppm, which correspond to C=N carbon.
The IR spectra of 4c showed an absorption peak at 3455 cm -1 which are responsible for ν(NH) stretching vibrations, while the absorption signal at 3081.54 cm -1 resulted from ν(CH) aromatic vibrations. The peak at 1687.18 cm -1 is responsible for absorbing the ν(C=O) stretching vibrations whereas two signals at 1629.75 and 1615.48 cm -1 results from ν(C=N) and ν(C=C) stretching vibrations of the aromatic ring. The sharp peak in the wavelength of 1552.32 cm -1 resulted due to asymmetric ν(NO2) stretching vibrations, while absorption signal at 1347.86 cm -1 reflects symmetric ν(NO2) stretching vibrations. A signal at 1204.76 cm -1 is characteristic for (C-O-C) stretching vibrations of the lactonic system, while the sharp peak at 764.53 cm -1 appeared from δ(C-H) bending oop vibrations of the aromatic ring. In the 1 H-NMR spectrum, the multiplet signals of aromatic protons appeared at 8.1-8.3 ppm and 7.7-7.4 ppm. A proton singlet resulting from N=C-H appears at 8.5 ppm. In the 13 C-NMR spectrum also displayed a signal at 163.7 ppm, that correspond to C=N and a signal at 162.6 ppm resulted from C=O carbon.
In the IR spectra of the compound 5a, a sharp absorption signal appeared at 3490 cm -1 , which is responsible for ν(NH) stretching vibrations whereas the absorption peak at 3086.34 cm -1 resulted from ν(CH) vibrations of the aromatic system. The medium band at 2933.88 cm -1 resulted from the absorptions of ν(CH) stretching vibrations of aliphatic protons, whereas the sharp peak at 1687.18 cm -1 from ν(C=O) stretching vibrations. The absorption signal at 1611.68 cm -1 resulted from ν(C=C) stretching aromatic vibrations. The characteristic signals of the nitro group appeared at 1516.35 cm -1 due to asymmetric stretching, whereas at 1328.68 cm -1 for symmetric ν(NO2) stretching vibrations. The absorption peak at 1206.66 cm -1 resulted from lactonic ν(C-O-C) stretching vibrations, whereas at 756.92 cm -1 appeared the absorption signal resulted from δ(CH) oop vibrations of the aromatic system. In the 1 H-NMR spectra, a proton singlet at 5.6 ppm resulted from N-C-H, while two doublets at 6.7 and 6.9 ppm and a multiplet at 7.5-7.0 ppm correspond to aromatic protons. A doublet at 3.5 ppm resulted from aliphatic CH2 protons of the azetidinone system. 13 C-NMR spectra appeared three peaks at 172.5, 162.6 and 161.8 which results from C=O and C-N carbons, whereas a signal at 55.4 ppm resulted from the CH-N of the azetididinone ring. An absorption signal at 36.4 ppm appeared due to methylene carbon.
In the IR spectra of the compound 5b, a broad absorption signal appeared at 3540-3280 cm -1 which are responsible for ν(OH) stretching vibrations and the absorption signal at 3111.04 cm -1 for ν(CH) stretching vibrations of the aromatic ring. The peak at 2942.48 cm -1 resulted from the absorptions ν(CH) stretching vibrations of the methylene group, while at the peak of 1721.68 cm -1 correspond to ν(C=O) stretching vibrations. The characteristic peak at 1669.12 cm -1 resulted from ν(C=C) stretching vibrations of aromatic moiety. Signals at 1544.06 and 1341.35 cm -1 appeared due to ν(NO2) asymmetric and symmetric stretching vibrations, whereas the characteristic signal at 1184.28 cm -1 is responsible for lactonic ν(C-O-C) vibrations. In the 1 H-NMR spectra are shown characteristic signals at 3.6 ppm (d, 2H, CH2), and at 5.7 ppm (t, 1H, N-C-H). 13 C-NMR spectra also showed a signal at 162.8 (C=N) and absorptions at 56.5 ppm (CH-N) and 37.2 ppm (CH2).

Table 2. Zones of inhibition (mm) of the discs impregnated with various concentration of the synthesized compounds
No.

S. aureus E. coli Klebsiella
Compound 5c appeared the absorption signal at 3433 cm -1 which is responsible for ν(NH) stretching vibrations and the absorption signal at 3078.38 cm -1 , which resulted from aromatic ν(CH) stretching vibrations. The peak at 2915.68 cm -1 resulted from ν(CH) stretching vibrations of the methylene group, while the signal at 1682.17 cm -1 absorption reflects the vibrations ν(C=O) stretching. The peak at the wavelength of 1628.55 cm -1 resulted from vibrations ν(C=C) stretching mode. The absorption peak at 1506.43 cm-1 resulted from ν(NO2) asymmetric stretching vibrations, while symmetric ν(NO2) stretching vibrations resulted by absorption peak at 1322.48 cm -1 . The absorption signal at 1215.25 cm -1 is characteristic for (C-O-C) stretching vibrations of the lactonic system, while the sharp peak at 708.19 cm -1 is characteristic for δ(C-H) oop bending vibrations of the aromatic ring. On the other hand, characteristic signals from the 1 H-NMR spectrum, a triplet at 5.8 ppm and a doublet at 3.7 pm correspond to proton absorptions of N-C-H and CH2 of the azetidinone ring. Also in the 13 C-NMR spectra, the azetidinone carbon signals appear at 54.6 ppm (CH-N) and 38.6 ppm, (CH2).

Antibacterial activity of the products 4a-4c and 5a-5c
Following this study, compounds 4a-4c and 5a-5c are investigated for their antibacterial activity. Our research is oriented to test the activity against bacteria S. aureus, E. coli, and Klebsiella, on the basis of Standard Disc Method, 24 by measuring the zones of inhibition around the standard discs. The discs have previously been impregnated with solutions of the products in DMF with concentrations of 2 mg mL -1 , 4 mg mL -1 and 6 mg mL -1 . Results are shown in Table 2.
Compounds of series 4 showed moderate antimicrobial activity against these microorganisms, whereas those of series 5 exhibited significant activity. Products 5b and 5c were most active against S. aureus, compounds 5b and 5a showed the most activity against E. Coli whereas 5a and 5b were more active against Klebsiella.
Antibacterial activity against E. Coli and Klebsiella appeared as bactericide activity is displayed in large-scale. Furthermore, these compounds expressed both bacteriostatic and bactericide activity against S. Aureus.   Bacteriostatic activity is exhibited in a large range (+2.0 mm), whereas bactericide activity showed in small diameter. Azetidin-2-one moiety showed significant impact on antimicrobial activity, whereas the impact of polar groups also was distinct. It is particularly noted the impact of the hydroxy group of 5b, which has affected the increase in antibacterial activity. Moreover, nitro group of 5b has shown significant impact on the range of inhibition of E. coli.
The assumption is that antibacterial activity may result as a consequence of the involvement of these products in enzymatic reactions. These products can cause enzymatic inhibition, by inhibiting cell wall construction of the microorganisms. However, mechanism of enzymatic inhibition is not yet fully studied. In general, by increasing the concentration of solvents, their antimicrobial activity increased.

Conclusions
New derivatives of 4-[4-(benzylidene-amino)phenylamino]-3-nitrobenzopyran-2-ones, 4a-4c and respective azetidin-2-ones, 5a-5c are synthesized in the moderate and high yield. Compounds 5b and 5c were most active against S. aureus, compounds 5b and 5a showed more activity against E. Coli, while 5a and 5b were more active against Klebsiella bacteria. The impact of polar groups in antibacterial activity was significant. Antibacterial activity is shown to be proportional to the concentration of these compounds.

INTRODUCTION
The problem of extraction of heavy metals from the sewage of mining production is very daunting in industrial regions. 1 In the process of long-term storage of the residues of processing of polymetallic sulfide ores, under the action of air oxygen, the gradual oxidation of sparingly soluble sulfides of heavy metals takes place with a formation of readily soluble compounds -sulfites and sulfates. These oxidation products penetrate to undersoil mine and shaft waters as a result of leaching with a consequent environmental contamination.
The ions of heavy metals, by a stress indicator, by their harmful environmental effect are even ahead of the residues of nuclear power plants and entities producing the organic compounds. Biosphere contamination by the ions of heavy metals is at the first place and is estimated as 135 points in the scale of ecological stress-factor, whereas an environmental contamination by radiation residues is estimated as 40 points by the same scale. 2 Therefore purification of the quarry waters (QW) from the ions of heavy metals is primarily considered as the ecological problem. From another hand, QW which contain a significant amount of valuable components, and in connection with their high cost, may be considered as a raw material for the production of a marketable product. Elaboration of profitable technology for their extraction from mentioned waters by substantial economic effect is a highly actual problem.
The problem of purification of QW from the ions of heavy metals is also actual for Bolnisi Region of Georgia. In this region QW contain toxic ions, an amount of which is significantly higher than limiting permissible concentration (LPC): Annually from quarry streams the following is removed (in tons): Cu -1290; Zn -430÷760; Fe -1100÷1500; Cd -4.34; SO4 2ions -4400. Therefore QW, containing the significant amount of useful components should be considered as the anthropogenic raw material, and at their complex processing, the considerable profit may be obtained besides the solving of a major ecological problem. 3. The copper ions are one of the primary sources of hydrosphere contamination by the ions of heavy metals. Purification of QW from copper ions in Join-Stock Company "Madneuli" (Georgia) is mainly carried out by cementation method which is slightly improved in technological viewpoint by the fact that a low-grade iron powder is used instead of steel chips and scrap. As a result, for deposition of 1 ton of copper powder more than 2 tons of iron precipitator is used, whereas the theoretical consumption comprises 0.9 tons. Obtained product involves a low-grade concentrate containing 50-60 % of copper and 15-20 % of iron. Separation of pure copper from mentioned concentrate is a very complicated problem and obtaining of any other product is practically impossible. The greater is an oxidized iron and large granules in the powder the more significant amount of iron passes to the solution by chemical dissolving, and the smaller amount of iron participates in copper precipitation. This process takes place in the practice of Join-stock Company "Madneuli". Moreover, the technology is oriented to water purification only from copper ions.
Direct electrolysis (cathodic precipitation) is among the most promising methods of purification of the quarry and industrial sewage from copper ions, allowing the metal extraction from aqueous solutions as a goal product. In this case, at the cathode, the pure copper powder is separated (it's processing to useful products isn't a complex problem) and at the anode a neutralization of toxic components, Therefore electrolysis method doesn't receive a widespread use for copper extraction from diluted solutions (because of low current efficiency and polarization and, hence, because of high voltage at electrolyzer). But the problem is entirely solvable at the creation of corresponding construction of electrochemical reactor which will allow the performance of so-called channel electrolysis, that is to say, an electrolysis at a high rate of fluid motion along the electrodes and in the conditions of limiting current density (Jk).
Over the years, our Institute has been working on the elaboration of such electrolyzer but the mentioned plants had certain disadvantages. 4,5 EXPERIMENTAL For copper extraction from diluted solutions ( [Cu]  1 gL -1 ) by high degree and high current efficiency we have elaborated the original construction of basic electrolyzer with a corresponding hydrodynamic regime (Fig. 1). 6,7 Electrochemical reactor involves the cylindrical body with a conic bottom. In reactor body, a cassette with a radial located fixed electrodes of definite shape is inserted. Manganese titanium dioxide is used as anode material, characterized by high durability in the conditions of longterm operation of an electrolyzer.  The anodes are enclosed in an acid-resistant bag to avoid the permanent recharge iron ions existing in water. As a result, the power consumpion reduces by 5-15 %. The plates from stainless steel were used as the cathodes. In cassette center, the mixer is located which directs a liquid flow between the electrodes by a high rate. The electrode shape is favorable for the increase of the distance between opposite electrodes from the center to the periphery, on one side. This fact allows the reducing of current density on the electrodes to the same direction. Simultaneously the depolarizer concentration is decreased from the center to the periphery. Thus an automatic control of the correspondence between concentration and current density takes place which is favorable for maintenance of product maximum current efficiency.
Extraction of electropositive metals is performed in the regime of limiting current. As a result, the spongy precipitate is formed. On the other hand, electrode shape is favorable for the conservation of liquid flow of circular motion. The reactor operates by the hydro-cyclone principle and liquid flow, moving along the electrodes by high rate, removes the cathode product and involves it to a circular motion on the inner surface of cylindrical reactor body. The particles of copper powder lose a rate and penetrate into the collector through the holes on reactor bottom. In the reactor, a significant improvement of the intensity of forced convection and solving the problem of removal of copper powder from the cathodes is attained.
The technological process is continuous, and a reactor is similar to an (electric) filter, in which its selective extraction takes place at the passing of the solution, contaminated by toxic copper ions. Precipitate dispersity increases the area of its surface significantly and decreases the real current density considerably on the electrode surface. The voltage on reactor clamps is no more than 2-3 V. Obtained productcopper contains only the oxygen as an impurity which is easily removed at product treatment in reducing medium.

RESULTS AND DISCUSSION
Cascade arrangement of two mentioned electrochemical reactors, one of which operates at high current densities and another at relatively low ones, gives 95 % copper extraction from QW (Table 2). Power consumption, in this case, comprises 4000 kWhourton -1 , corresponding to 5-10 % of the copper cost. At remelting of electrolytic copper powder, a copper ingot of 99.0 % was obtained For complex processing of QW for the purpose of extraction of useful components and their further processing for obtaining of valuable, marketable products, we have elaborated several technological schemes (Fig. 2).
Considering QW as the technogenic raw material the following marketable -commercial products may be obtained by mentioned schemes: copper ingots of 99.8 %; vitriol and other chemical compounds; bio dye and other pigments; zinc and its powder; gold; pure secondary water. O-optimistic estimation; P-pessimistic estimation; *in ounce, #1and #2; short and #3and #4 are full technologies. The prices for corresponding metals are taken from official data of London Exchange on 14.02.2017, on other productsby corresponding tariffs. Iinvestment, Pr-product, PA product amount in ton, MC-maintenance cost  The majority of these products are scarce. Therefore their realization is beyond question. When it is considered that the main raw material -QW is free of charge and field cost are relatively small, the price cost of obtained production is low. This fact increases its competitive ability significantly.
The scheme of processing of electrolytic copper is best suited for obtaining of vitrioltraditional fungicide for Georgian agriculture. Vitriol preparation was performed by conventional, but significantly simplified technology. Obtained vitriol contains a negligible impurity and is characterized by dendrite structure with developed surface.
In Table 3 the estimation of economic efficiency of QW processing by proposed schemes is presented. In the case of full-scale realization of proposed technology annually obtaining of the profit 8-9 million US $ is possible Table 3.

CONCLUSIONS
1. Construction of electrochemical reactor has been elaborated allowing to perform the process of purification of quarry waters from copper ions by high efficiency (copper extraction by 95 %) at power consumption corresponding to 5±10 %.
2. Technological schemes were elaborated for complex processing of quarry waters for separation of useful components for their processing into costly marketable production.
3. It was shown that at a full-scale realization of elaborated technological schemes an annual profit of 8-9 million US dollars might be obtained. 4. The hydrometallurgical technology of sewage purification of our elaboration belongs to so-called selffinancing technologies. Even insignificant investment initiates a self-financing.