1. 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).

2. 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).

3. 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.

4. 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

5. 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.

6. ANTIMICROBIAL ACTIVITY
• Microorganisms used:
gram positive bacteria viz:Staphylo
cocus aureus and Bacillus subtilus and S.N
Ingredients Weight(g)
O
gram negative bacteria viz: Escherichia coli
• Preparation Of Inoculum: 1.
Beef extract 4.0
The suspensions of all the
organisms were prepared as per Mac- 2.
Peptone 5.0
Farland Nephelometer standard(Baily
and Scott 1990). A 24hr old culture was 3.
Agar 20.0
used for the preparation of bacterial
suspension. Suspensions of organisms 4.
Distilled water q.s 1000ml
were made in sterile isotonic solution of
sodium chloride (0.9 %W/V) and the 5.
Ph 5.4
turbidity was adjusted.
• udomonus aeruginosa species.

7. 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 4C for 15 minutes for
diffusion to take place. After diffusion , the petridishes were incubated at 37c 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.

8. ANTIFUNGAL ACTIVITY
FUNGI USED:
S.no Ingrediants Weight in g
• Standard cultures of Candida
albicans and Aspergillus niger. 1. Dextrose 40
NUTRIENT MEDIUM
• Dextrose,Peptone,Agar 2.
Peptone
10
Distilled water, These ingredients
3. Agar 20
were accurately weighed and
dissolved water.The medium Is
4. Distilled water q.s 1000ml
prepared was sterilized by
autoclaving at 121C for 15 minutes

9. • 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

10. RESULTS
Sl. NO STRUCTURE ZONE OF INHIBITION IN MM
S.a B.s E.c P.a
1. 2–imino-8–benzylidene-4–
phenyl5,6,7–trihydroxy-4H,7H– 21 18 15 17
(3,1)benzoxazine
CONTROL DMSO - - CONTROL DMSO
STD Ampicillin 19 17 STD Ampicillin

11. 6. DISCUSSION
• 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).

12. 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-
methyl benzylidene)-4[methyl phenyl]-5,6,7-trihydro-4H,7H-
(3,1)benzoxazine[II(2)] and 2-imino-8-(4-chioro benzylidene)-4-[4-
chloro]-5,6,7-trihydro-4H,7H-(3,1)benzoxazine II(4) has shown highest
zone of inhibition indicating that introduction of groups such as –CH3,-Cl
in II (1) enhances the activity against Staphylococcus aureus.

13. ANTIFUNGAL ACTIVITY:
STRUCTURE ACTIVITY RELATIONSHIPS
• The standard reference drug was Amphotericin B
• The zone of inhibition shown by the compound 2-imino-8-
benzylidene-4-phenyl-5,6,7-trihydro-4H,7H-(3,1)benzoxazine.
II(1) aganist Candida albicans is 13mm.
• Among all the derivatives the compound 2-imino-8-(4-
chiorobenzylidene)-4-(4-chlorophenyl)-5,6,7-trihydro-4H,7H-
(3,1)benzoxazine II(4) had shown highes zone of inhibition
indicating that introduction of chloro substituent in II(1)
enhances the activity.

14. CONCLUSSION & SUMMARY
• The aim of our present study was to synthesize,characterize and to evaluate
antibacterial and antifungal activity of some oxazines was completed successfully.
• The intermediate compounds, 2,6-diarylidene-cyclohexanone derivatives I(1-4)
have been prepared by condensation of one mole of cyclohexanone with two moles of aromatic
aldehyde.
• The final compounds,2-imino-8-arylidene-4-aryl-5,6,7-trihydro-4H,7H-(3,1)benzoxazine
derivatives II(1-4) have been prepared by cyclo condensation of I(1-4) with urea.
• The structure of some synthesized compounds have been authentically established by
their,UV,IR spectral studies.
• The antibacterial activity of II (1-4)has been evaluated. The anti fungal activity of II (1-4) has
been evaluated.

15. THANKING
YOU
SIR…