SYNTHESIS AND ANTI MICROBIAL EVALUATION OF SOME OXAZI DERIVATIVES SEMINAR BY G.VIJENDHAR (08GD1R0007) M HIMA BINDU (08GD1R0014) K. RACHANASREE (08GD1R0023) M.PRADEEP KUMAR (08GD1R0024) M.MANISHA (08GD1R0047) CH. MAHENDAR REDDY (08GD1R0050) UNDER THE GUIDENCE OF V.KALYAN VARMA, M.Pharm. Department of Pharmaceutical Chemistry CHILKUR BALAJI COLLEGE OF PHARMACY (SPONSORED BY SRINIVASA EDUCATIONAL ACADEMY, CHITTOOR) (APPROVED BY AICTE,PCI, NEW DELHI) (Affiliated to Jawaharlal Nehru Technological University, H
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SYNTHESIS AND ANTI MICROBIAL EVALUATION OF SOME OXAZINE
DERIVATIVES SEMINAR BY
G.VIJENDHAR (08GD1R0007) M HIMA BINDU (08GD1R0014) K. RACHANASREE (08GD1R0023) M.PRADEEP KUMAR (08GD1R0024) M.MANISHA (08GD1R0047)
CH. MAHENDAR REDDY (08GD1R0050)
UNDER THE GUIDENCE OF
V.KALYAN VARMA, M.Pharm.
Department of Pharmaceutical Chemistry
CHILKUR BALAJI COLLEGE OF PHARMACY (SPONSORED BY SRINIVASA EDUCATIONAL ACADEMY, CHITTOOR)
(APPROVED BY AICTE,PCI, NEW DELHI) (Affiliated to Jawaharlal Nehru Technological University, Hyderabad, A.P).
INTRODUCTION
• 1,3-Oxazines are a well-known structural modify for biologically active substances. They exhibit antifungal, antibiotic, antiviral, and antitumor properties and are also used as acetylcholinesterase (AChE) inhibitors; potential targets for Alzheimer’s drugs.
• The biological activity of oxazine derivatives was reported as early as 1937 (Novelli& Adams, 1937).Later, several workers reported the fungistatic and bacteriostatic-including tuberculostatic-activity of these compounds (Urbanski& Slopek, 1951; Lane, 1953; Kay &Lane, 1953; Shono&Takahashi, 1954).
OBJECTIVES• 1,3 oxazines derivatives are an interesting class of
heterocyclic compounds being studied in recent years and are reported to posses a wide spectrum of biological activities such as antibacterial, antifungal, anti-inflammatory, anti-helminthec, antitumor and anti HIV activities.
• Many heterocyclics have shown antifungal activity due to N-C=O-N group and this group is also present in the structure of 1,3-Oxazine nucleus.
• The above observations stimulated us to synthasise more number of useful therapeutic derivatives having Oxazine nucleus .
• 1.synthesis and evaluation of Oxazine derivatives.• 2.evaluation of the antimicrobial activities like antibacterial
and antifungal activity of some Oxazine derivatives.
METHODOLOGY
A mixture of 30ml of 10% sodium hydroxide ,
50ml of ethanol and 0.01mol of cyclohexanone
placed in a 250 ml beaker ,which was
immersed in ice bath as well as equipped with
a mechanical stirrer ,The stirrer was started
0.02ml of benzaldehyde was added to the
mixture and stirring continued After 2 hours,
the stirrer was removed and the reaction
mixture was kept in an ice chest over night.
The product was filtered, washed with ice cold
water , followed by ice cold ethanol , dried and
recrystallized from DMF
METHODOLOGY-2
• A mixture of 0.01mol 2,6 –dibenzylidene
cyclohexanone of 0.015mol of urea and
0.01ml of potassium hydroxide dissolved
in 10ml of water was reflexed in ethanol
for 10 hours. Later ethanol was removed
under reduced pressure and the residue
was treated with ice cold water .The
precipitate thus obtained was filtered
washed with water,dried and
recrystallized from ethanol.
ANTIMICROBIAL ACTIVITY
• Microorganisms used:
gram positive bacteria viz:Staphylo
cocus aureus and Bacillus subtilus and gram
negative bacteria viz: Escherichia coli
• Preparation Of Inoculum:
The suspensions of all the organisms
were prepared as per Mac-Farland
Nephelometer standard(Baily and Scott
1990). A 24hr old culture was used for
the preparation of bacterial suspension.
Suspensions of organisms were made in
sterile isotonic solution of sodium
chloride (0.9 %W/V) and the turbidity
was adjusted.
• udomonus aeruginosa species.
S.NO Ingredients Weight(g)
1.Beef extract 4.0
2.Peptone 5.0
3.Agar 20.0
4.Distilled water q.s 1000ml
5.Ph 5.4
PROCEDURE:
• Inoculum was added to 30ml of sterile nutrient agar medium and was poured into
sterile petridishes for solidifying.
• 0.1ml of test solution was added to the petridishes, 0.1ml of the ampicillin at a
concentration of 100μg/ml was taken as standard reference.
• The petridishes were kept in the refrigerator at 4 ۫C for 15 minutes for diffusion to
take place. After diffusion , the petridishes were incubated at 37 ۫c for 24hr and zone of
inhibition were observed and measured using a scale.
• Antibacterial activity of all the compounds was carried out against all four
microorganisms. The same media was used for both sub culturing and for estimating
antibacterial activity.
ANTIFUNGAL ACTIVITYFUNGI USED:
• Standard cultures of Candida
albicans and Aspergillus niger.
NUTRIENT MEDIUM
• Dextrose,Peptone,Agar
Distilled water, These ingredients
were accurately weighed and
dissolved water.The medium Is
prepared was sterilized by
autoclaving at 121 ۫C for 15 minutes
S.no Ingrediants Weight in g
1. Dextrose 40
2.Peptone
10
3. Agar 20
4. Distilled water q.s 1000ml
• WORKING PROCEDURE:
• An inoculum was prepared by suspending a single isolated colony in above 5ml of normal saline. Later one drop of tween 20 was added for filamentous fungi and the mouid was broken by shaking.
• A sterile cotton swab was moistened in the inoculum suspension and excess of moisture was removed by rolling the cotton swab on the inside of the tube, above fluid level 30ml of sterile hot sabauads agar medium was poured in each plate and allowed to harden on a level surface of sabouraud's agar medium plate was streaked with the help of moistened cotton swab in all the direction ions.
• The surface of plate was dried out 35°C. Later 5 bores per plate were made using sterile cork bores.
• The operation was carried out in asceptic condition and 0.1ml test solution was added to the respective bore and 0.1ml Amphotercin was taken as standard reference. A control having only DMSO was maintained in each plate. The plates are incubated at 35°C for 48hrs. Later the values of zones of inhibition were recorded
• The structure of new compounds prepared during the present investigation have been authentically established by their UV,IR spectral studies.In the following the spectral studies of some selected compounds have been detailed.
• The compound 2,6-dibenzylidine-cyclohexanone[I(1)]has been prepared
by condensing 1mol of cyclohexanone with 2moles of benzaldehyde the for I(1)as
been indicated by its uv spectrum.the starting material cyclohexanone exhibited
λmax at 287nm.the compoundsI(1) exhibited λmax at 328nm.this clearly indicates
that the bathochromic shift may be attributed because of =CH-Ar chromophore.
• The formation of I(1)as been indicated by its IR spectrum the starting
material exhibited γmax at 1661cm-1 due to C=O group.the compoundsI(1) exhibited
γmax at 1607cm-1 due to C=Ogroup.the appearance of a characteristic band at C=O is
mainly due to α,β,and α-1 β-1.this clearly indicates the formation of I(1).
ANTIBACTERIAL ACTIVITY:
STRUCTURE ACTIVITYRELATIONSHIPS
• The zone of inhibition shown by the compound 2-imino-8-benzylidene-
4-phenyl 5,6,7-trihydro-4H-7H-(3,1)benzoOxazine[II(1)] against
Staphylococcus aureus is 21nm.
• Among all the compounds, two derivatives viz.2-imino-8(4-
