chapter iv result and discussion -...

ChapterIV

Result and discussion

Suthar Arunkumar G. Page 144

Chapter-IV

** RESULT AND DISCUSSION **

In this chapter the result obtained during the course of present investigation have been discussed

under the following section:

SECTION-I: SPECTROSCOPIC ANALYSIS OF THE SYNTHESISED

COMPOUNDS

We have determined the structures of the synthesized compounds by their Infrared (IR), Proton

Magnetic Resonance (PMR) and Mass Spectra.

IR Spectroscopy

Introduction:

Infrared spectroscopy is one of the most common spectroscopic techniques used by organic and

inorganic chemists. Simply, it is the absorption measurement of different IR frequencies by a

sample positioned in the path of an IR beam.

The main goal of IR spectroscopic analysis is to determine the chemical functional groups in the

sample. Different functional groups absorb characteristic frequencies of IR radiation. Using

various sampling accessories, IR spectrometer can accept a wide range of sample types such as

gases, liquids and solids. Thus, IR spectroscopy is an important and popular tool for structural

elucidation and compound identification.

IR absorption information is generally presented in the form of a spectrum with wavelength or

wave number and absorption intensity or percent transmittance. The IR region is commonly

divided into three smaller areas. Near IR, middle IR and far IR. We have focused on the most

frequently used idle IR region between 4000-400 cm-1.


PMR Spectroscopy

General uses:

(a) Identification and proof of structure of chemical compounds.

(b) Quantitative determination of sample compounds

(c) Detailed information on the spatial orientation of nuclei in a molecule.

(d) Studies of dynamic systems including chemical equilibria,molecular motion and

intermolecular interactions.

Common applications:

(a) Widely applied for structure proof in synthetic chemistry, natural products chemistry

and biochemistry.

(b) Determining stereochemistry and higher-order structures in molecules of all sizes.

(c) Conformational analysis, including assessment of mixtures in conformational equilibria.

(d) Following the course of chemical reactions by identifying and quantifying the starting

materials and products.

(e) Intermolecular interactions such as ion-pair formation or enzyme-substrate interactions.

(f) The study of chemical exchange, such as valence bond tautomerism.

Introduction:

In 50 years, since the first observation of nuclear/proton magnetic resonance (NMR), the

technique has become an indispensable tool for the chemists. In the early years, applications of

proton NMR were quickly extended to structural studies of all major types of organic

compounds. The utility of NMR in the study of dynamic chemical systems and in conformational

analysis has also become clear. The development of powerful superconducting magnets and the

introduction of the pulsed Fourier transform (FT) technique vastly increased the sensitivity and

resolving power of the method.

The sensitivity improvement meant that NMR could be extended to nearly all elements in

the periodic table. Today’s NMR spectrometer incorporates the advances of the last 50 years in a


versatile, easy to use instrument. Automated data acquisition and processing provide access to

the most useful experiments on a routine basis.

The practical application of NMR spectroscopy to the study of the structure of complex

compounds became possible only after the discovery in 1951 that the spectrum of ethanol

consists of three individual signals corresponding to the resonance of the methyl, methylene and

hydroxy protons and that the signals of different groups of magnetic nucleus in liquid molecules

give rise to further fine splitting which depends on the number and character of the nuclei

contained in the molecule. Since the nucleus involved here for ethanol was the proton the

spectrum is some time called Proton Magnetic Resonance (PMR) spectrum, to differentiate it

from spectra involving such nuclei as 13C, 14N or 19O.

Mass Spectroscopy

General uses:

(a) Identification of organic compounds by virtue of fragmentation pattern.

(b) Determination of molecular weight.

(c) Elucidation of structural features in an unknown molecule.

(d) Determination of isotope incorporation of stable isotopes.

(e) Estimation of elemental composition based on appearance and multiplicity of

isotope peaks.

Common applications:

(a) Identification of common environmental contaminants.

(b) Identification of drugs and drug metabolites.

(c) Verification or conformation of product or products of organic synthesis.

(d) Determination of the extent of incorporation of stable isotope-labeled

tracer compound into biological pool of hormones or other chemical

transmitters.


(e) Determination of the kinetics of biosynthesis or organic chemical

turnover as a precursor or as a metabolite in a biological system.

Introduction:

Mass spectroscopy was originally designed by physicists at the turn of the century to

determine the mass-to-charge ratio (m/z) of charged particles by virtue of their behavior in

magnetic and electric fields. The early interest of the physicists was in proving existence of

isotopes of various elements. Organic mass spectrometry was cultivated by scientists in the

petroleum industry starting in the 1940s. Mass spectrometry expanded into biomedical

applications in the 1970s. In general, mass spectrometry is a micro analytical technique that

requires some energetic process for converting a significant number of molecules of the analyze

to a charged species so that the (m/z) ratio of the charged form of the analyze may be

determined. A given compound is converted to a charged species by one of the variety of

available ionization processes.

A mass spectrometer bombards the substance under investigation with an electron beam

and quantitatively records the result as a spectrum of positive ion fragments. This record is a

mass spectrum. Separation of the positive ion fragments is on the basis of mass (strictly,

mass/charge, but the majority of ions are singly charged).


**IR Spectra**

IR spectral study of 2-(Adamantan-1-yl-{4-[4-(benzylidene-amino)-phenyl]-thiazol-2-yl}-

amino)-acetamide (ASI-01)

Figure -1

Sample technique : KBr Pellet Instrument : Perkin Elmer Spectrum 100 Frequency range : 4000-400 cm-1

Type Vibration mode Frequency cm-1 Amine -N-H Str. of –CONH 3426.6 Alkyl C-H Str. Symm. 2888.3 Amide -C=O str. 1656.8

Nitrile -CN Str. 1356.8

Aromatic

>C=C ring skeleton vib. 1578.5 1535.2

o-subst. bending vib Mono subst. bending vib

737.6 705.7


IR spectral study of 2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-phenyl-

acetamide (ASII-01)

Figure -2


Type Vibration mode Frequency cm-1 Amine -N-H Str. of –CONH 3409.3 Alkyl C-H Str. Symm. 2832.9 Amide C=O str. 1690.2


Aromatic

>C=C ring skeleton vib. 1514.3 1552.7 1608.1

Aromatic -C-H Str. 3068.5



amino)-N-propyl-acetamide. (ASIII-01)

Figure -3


Type Vibration mode Frequency cm-1 Amine -N-H Str. of –CONH 3411.1 Alkyl C-H Str. Symm. 2941.4 Amide -C=O str. 1690.0


Aromatic




amino)-N-phenyl-acetamide. (ASIV-01)

Figure -4

Sample technique : KBr Pellet

Instrument : Perkin Elmer Spectrum 100

Frequency range : 4000-400 cm-1 Type Vibration mode Frequency cm-1

Amine -N-H Str. of –CONH 3319.54 Alkyl -C-H Str. Symm. 2927.92 Amide -C=O str. 1694.40


Aromatic



IR spectral study of 2-((4-Phenyl-thiazol-2-yl)-{4-[2-(piperidine-1-carbonyl)-phenoxy]-phenyl}-

amino)-acetamide (ASv-01)

Figure: 5 Sample technique : KBr Pellet Instrument : Perkin Elmer Spectrum 100 Frequency range : 4000-400 cm-1

Type Vibration mode Frequency cm-1

Amine -N-H Str. of -CONH 3410 Alkyl

C-H Str. Symm.

C-H Str. Asymm. 2930 2860

Amide C=O str. 1698

Aromatic >C=C ring skeleton vib. 1577,1524,1455,1381


756 744

Ether C-O str. (diakyl ether) 1269


IR spectral study of N-Phenyl-2-[5-Phenyl-3-(1-Phenyl-3,4-Dihydro-1H-Isoquinolin-2-yl)-

[1,2,4]Triazole-1-yl]-Acetamide (ASVI-01)

Figure -6



Amine -N-H Str. of –CONH -N-H Str. of -NH

3420 3310

Alkyl -C-H Str. Symm. 2938 Amide -C=O str.

1678 1570

Aromatic

>C=C ring skeleton vib. 1560 1608 1533


755 705


IR spectral study Benzylidene-{4-[2-ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-2H-

[1,2,4]triazol-3-yl]-phenyl}-amine of (ASVII-01)

Figure: 7 Sample technique : KBr Pellet Instrument : Perkin Elmer Spectrum 100 Frequency range : 4000-400 cm-1


Alkyl

C-H Str. Symm. C-H Str. Asymm.

2928.9 2852.5

Alkene =C-H Str medium 3072.5 =C-H Str strong 807.18 C=C Str variable 1667.8

Imine C=N str 1628.7 Amine C-N str medium-weak 1214.00 Nitrile CN Str medium 2238.6

Aromatic

>C=C ring skeleton vib. 153.2,1543,1442,1366.7 p-subst. bending vib


833.14 727.76

687.23


IR spectral study of N-{4-[2-Ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-2H-

[1,2,4]triazol-3-yl]-phenyl}-benzamide (ASVIII-01)

Figure -8



Amine -N-H Str. of –NH

3278.3 3353.9

Amine -N-H banding. of -NH 1618.5

Alkyl C-H Str. Symm. 2940.3 Amide C=O str. 1653.9

Aromatic

>C=C ring skeleton vib. 1495.9,1593.3,1618.5 o-subst. bending vib p-subst. bending vib

839.7 748.7

Aromatic

C-H Str. Symm 3093.8


IR spectral study of N-(1-Cyano-1,4-dihydro-cyclopenta[c]chromen-2-yl)-benzamide (I) (ASIX-

01)

Figure -9



Frequency range : 4000-400 cm-1

Type Vibration mode Frequency cm-1 Amine -N-H Str. of –CONH 3463.43 Alkyl C-H Str. 2927.70 Amide C=O str. 1686.62

Ether C-O str. 1260.62



Suthar Arunkumar G. 7

IR spectral study of N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-

METHANESULFONAMIDE (ASX-01)

Figure -10




Type Vibration mode Frequency cm-1 Amine -N-H Str 3419.62 Alkyl C-H Str. 2927.92 Amide S=O str. 1351.50

Ether C-O str. 1185.9




IR spectral study of 2-(BENZYLIDENE-AMINO)-1,4-DIHYDRO-

CYCLOPENTA[C]CHROMENE-1-CARBONITRILE(ASXI-01)

Figure -11




Type Vibration mode Frequency cm-1 Alkane C-H Str. 2927.92 Nitrile -CN Str. 2256.9

Aromatic -C-H Str. 3149.5 Aromatic -C=C Str. 1508.9

1541.8 1598.1

Ether -C-O Str. 1152.3


IR spectral study of 2-(5-Benzylidene-4-oxo-2-phenyl-thiazolidin-3-yl)-1,4-dihydro-cyclopenta[c]chromene-1-carbonitrile(ASXII-01)

Figure -12




Type Vibration mode Frequency cm-1 Alkane C-H Str. 2927.50 Nitrile -CN Str. 2218.61

Aromatic -C-H Str. 3110.91 Ketone -C=O Str. 1775.43

Aromatic -C=C Str. 1507.59 1599.51

Ether -C-O Str. 1159.76


**NMR Spectra**

NMR spectra of 2-(Adamantan-1-yl-{4-[4-(benzylidene-amino)-phenyl]-thiazol-2-yl}-amino)-

acetamide (ASI-01)

Figure: 13 Instrument : Varian 400 MHz FT-NMR Standard : TMS Solvent : DMSO d6

δ ppm Multiplicity No. of protons Inference 1.17-1.57 m 15H 3CH2

,3CH(a,b,c,d,e,f,g,h,i,j) 4.22 s 2H CH2 (k) 4.68 s 1H CH(m)

7.25-7.31 m 3H ArH(t,u,v) 7.38-7.41 d 4H ArH (n,o,s,w)

7.55-7.68 d 2H ArH (p,q)

8.15 s 1H =CH(r) 8.36 s 2H CONH2(l)


NMR spectral study of 2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-phenyl-

acetamide (ASII-01)



,3CH(a,b,c,d,e,f,g,h,i,j) 4.28 s 2H CH2 (k) 4.77 s 1H CH(l) 6.49 s 2H ArNH2(q)

6.78 dd 2H ArH(o,p) 7.20-7.23 m 3H ArH(u,v,w) 7.25-7.26 dd 2H ArH (m,n)

7.55-7.61 dd 2H ArH (s,t)

8.33 s 1H CONH(r)


NMR spectral study of 2-(Adamantan-1-yl-{4-[4-(benzylidene-amino)-phenyl]-thiazol-2-yl}-




,3CH(a,b,c,d,e,f,g,h,i,j) 1.89-1.92 dt 4H 2CH2(x,y) 3.15-3.19 dt 4H 2CH2(w,z) 4.31 s 2H CH2 (k) 4.75 s 1H CH(l)

7.21-7.30 m 3H ArH(s,t,u) 7.35-7.40 d 4H ArH (r,v,n,m) 7.48-7.55 d 2H ArH (o,p)

8.13 s 1H =CH(q)


NMR spectral study of 2-(Adamantan-1-yl-{4-[4-(benzylidene-amino)-phenyl]-thiazol-2-yl}-




,3CH(a,b,c,d,e,f,g,h,i,j) 3.19-3.21 s 2H CH2(r) 4.12-4.14 s 1H CH(q) 4.18-4.20 s 2H CH2(k) 4.72-4.74 s 1H CH(l) 6.92-6.94 t 2H ArH (v,w) 7.11-7.16 d 2H ArH (s,t) 7.26-7.32 m 6H ArH(o,p,u1,u2z1,z2) 7.61-7.73 dd 4H ArH (m,n,y1,y2) 1.89-1.92 dt 4H 2CH2(x,y) 10.18 s 1H CONH(x)


NMR spectral study of 2-((4-Phenyl-thiazol-2-yl)-{4-[2-(piperidine-1-carbonyl)-phenoxy]-

phenyl}-amino)-acetamide (ASV-01)


δ ppm Multiplicity No. of protons Inference 3.11-3.25 t 2H CH2 (a) 3.99-4.05 t 2H CH2 (b) 4.96-4.99 s 1H CH(c) 5.22 s 2H CH2 (d) 3.45-3.68 m 8H CH2 (e,f,g,h) 6.77-6.80 t 1H Ar-H (i)

6.93-7.01 m 2H ArH (j,k) 7.05-7.10 m 2H ArH (l,m) 7.11-7.19 m 2H ArH (n,o) 7.54-7.57 t 2H ArH (p,q) 7.68-7.85 m 3H ArH (r,s,t) 7.87-7.94 d 2H ArH (u,v)


NMR spectral study of N-Phenyl-2-[5-Phenyl-3-(1-Phenyl-3,4-Dihydro-1H-Isoquinolin-2-yl)-

[1,2,4]Triazole-1-yl]-Acetamide (ASVI-01)


δ ppm Multiplicity No. of protons Inference 3.16-3.18 t 2H CH2 (a) 3.97-4.12 t 2H CH2 (b) 4.93-5.03 s 1H CH(c) 5.13 s 2H CH2 (d) 6.78-6.86 m 1H ArH (e)

7.08-7.18 m 3H ArH (f,g,h)

7.19-7.23 d 2H ArH (i,j) 7.32-7.41 t 2H ArH (k,l) 7.55-7.60 m 7H ArH (m,n,o,p,q,r,s) 7.81-7.97 m 4H ArH (t,u,v,w) 10.56 bs 1H ArH (x)


NMR spectral study of Benzylidene-{4-[2-ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-

2H-[1,2,4]triazol-3-yl]-phenyl}-amine of (ASVII-01)


δ ppm Multiplicity No. of protons Inference 1.36-1.44 t 3H CH3 (e) 3.02-3.05 t 2H CH2 (a) 3.91-3.93 t 2H CH2 (b)

4.08-4.10 q 2H CH2 (d) 4.98-5.03 s 1H CH (c) 6.96-7.03 t 3H ArH(f,g,i) 6.79-6.84 m 1H ArH (h) 7.13-7.16 d 2H ArH (j,k)

7.29-7.36 m 4H ArH (l,m,r,s)

7.33-7.48 m 3H ArH (o,p,q) 7.55-7.68 m 4H ArH (t,u,v,w) 8.12-8.17 s 1H ArH (x)


NMR spectral study of N-{4-[2-Ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-2H-



δ ppm Multiplicity No. of protons Inference 1.35-1.43 t 3H CH3 (e) 3.03-3.06 t 2H CH2 (a) 3.92-3.94 t 2H CH2 (b)

4.05-4.07 q 2H CH2 (d)

4.99-5.02 s 1H CH (c) 6.98-7.02 t 3H ArH(f,g,i) 6.81-6.85 t 1H ArH (h) 7.12-7.14 d 3H ArH (j,k,n)

7.18-7.23 m 2H ArH (i,m)

7.58-7.61 m 5H ArH (o,p,q,t,u) 7.79-7.85 t 2H ArH (r,s) 7.99-8.02 m 2H ArH (v,w) 8.15 s 1H -NH (x)


NMR spectral study of N-(1-Cyano-1,4-dihydro-cyclopenta[c]chromen-2-yl)-benzamide (I)

(ASIX-01)


δ ppm Multiplicity No. of protons Inference 3.64-3.66 s 1H CH(g) 4.5-4.52 s 2H CH2(e) 5.68-5.70 s 1H CH(f) 6.80-6.82 t 2H ArH(b,d)

7.08-7.10 t 1H ArH(c)

7.14-7.16 d 1H ArH(a)

7.42-7.47 m 3H ArH(k,l,m)

7.85-7.88 dd 2H ArH(i,j)

8.18-8.20 s 1H NH(h)


NMR spectral study of N-(1-Cyano-1,4-dihydro-cyclopenta[c]chromen-2-yl)-

methanesulfonamide (ASX-01)


δ ppm Multiplicity No. of protons Inference 2.77-2.79 s 3H CH3(i) 3.67-3.79 s 1H CH(g) 4.52-4.54 s 2H CH2(e) 5.64-5.66 s 1H CH(f) 6.75-6.79 t 2H ArH(b,d)

7.04-7.06 t 1H ArH(c)

7.11-7.13 d 1H ArH(a)

2.07-2.08 s 1H NH(h)


NMR spectral study of N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-

METHANESULFONAMIDE (ASXI-01)


δ ppm Multiplicity No. of protons Inference 3.79-3.81 s 1H CH(g) 4.57-4.59 s 2H CH2(e) 5.94-5.96 s 1H CH(f) 6.75-6.77 t 2H ArH(b,d)

7.08-7.10 t 1H ArH(c)

7.29-7.31 d 1H ArH(a)

7.47-7.49 m 3H ArH(k,l,m)

7.65-7.67 dd 2H ArH(i,j)

8.19-8.22 s 1H CH(h)


NMR spectral study of 2-(5-Benzylidene-4-oxo-2-phenyl-thiazolidin-3-yl)-1,4-dihydro-cyclopenta[c]chromene-1-carbonitrile(ASXII-01)


δ ppm Multiplicity No. of protons Inference 3.85-3.87 s 1H CH(g) 4.54-4.56 s 2H CH2(e) 5.78-5.80 s 1H CH(f) 5.86-5.88 s 1H CH(h) 6.09-6.11 s 1H CH(i) 6.72-6.74 t 2H ArH(b,d)

7.03-7.10 m 4H ArH(c,o,p,r)

7.12-7.18 m 3H ArH(a,q,s)

7.28-7.33 m 3H ArH(n,l,m)

7.45-7.48 dd 2H ArH(k,j)


**MASS Spectra**

MASS spectral study of 2-(Adamantan-1-yl-{4-[4-(benzylidene-amino)-phenyl]-thiazol-2-yl}-

amino)-acetamide (ASI-01)

Figure: 25

Sr. No. M.W M.F Mass (m/z)

ASI-01 470.86 C28H30N4OS 471.60 (M+1)


MASS spectral study of 2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-

phenyl-acetamide (ASII-01)

Figure: 26


ASI-01 458.65 C28H30N4OS 459.53 (M+1)




Figure: 27


ASIII-01 524.88 C32H36N4OS 525.80(M+1)




Figure: 28


ASIV-01 620.2 C36H36N4O2S2 621.26 (M+1)


MASS spectral study of 2-((4-Phenyl-thiazol-2-yl)-{4-[2-(piperidine-1-carbonyl)-phenoxy]-

phenyl}-amino)-acetamide (ASV-01)

Figure: 29


ASI-01 479.12 C29H29N5O2 480.03 (M+1)


MASS spectral study of N-Phenyl-2-[5-Phenyl-3-(1-Phenyl-3,4-Dihydro-1H-Isoquinolin-2-yl)-

[1,2,4]Triazole-1-yl]-acetamide (ASVI-01)

Figure: 30


ASII-01 485.58 C31H27N5O 486.30 (M+1)


MASS spectral study of Benzylidene-{4-[2-ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-

2H-[1,2,4]triazol-3-yl]-phenyl}-amine (ASVII-01)

Figure: 31


ASIII-01 483.61 C32H29N5 484.50 (M+1)


MASS spectral study of N-{4-[2-Ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-2H-


Figure: 32


ASIV-01 499.61 C32H29N5O 500.51 (M+1)


MASS spectral study of N-(1-Cyano-1,4-dihydro-cyclopenta[c]chromen-2-yl)-benzamide

(ASIX-01)

Figure: 33


ASI-01 314.66 C28H30N4OS 315.43(M+1)


MASS spectral study of N-(1-Cyano-1,4-dihydro-cyclopenta[c]chromen-2-yl)-

methanesulfonamide (ASX-01)

Figure: 34


ASII-01 288.45 C28H30N4OS 289.38 (M+1)


MASS spectral study of N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-

METHANESULFONAMIDE (ASXI-01)

Figure: 35


ASIII-01 298.44 C28H30N4OS 299.30(M+1)


MASS spectral study of 2-(5-Benzylidene-4-oxo-2-phenyl-thiazolidin-3-yl)-1, 4-dihydro-cyclopenta[c]chromene-1-carbonitrile (ASXII-01)

Figure: 36


ASXII-01 460.86 C29H20N2O2S 461.73 (M+1)


SECTION-II : BIOLOGICAL EVALUTION

ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF THE

COMPOUNDS SYNTHESISED IN CHAPTER-III

Introduction:

Legionnaire’s disease, influenza, jaundice, tuberculosis, typhoid, dermatomycoses, dysentery,

malaria etc. are human diseases caused by microorganisms (bacteria and fungi). Animals

(infected with brucellosis, tularemia, etc.) and plants (infected with mildews, rusts, smuts,

cankers, leaf spots, etc.) have also been known to be victims of microbial pathogens. So far as is

known, mankind is closely influenced by the activities of microorganisms. Microorganisms are a

part of our lives in more ways them most of us understand. They have shaped our present

environment and their activities greatly influence our future. Microorganisms should not be

considered separate from human beings, but should be considered as a part of our life.

Microorganisms have a profound beneficial influence on our daily life. They are employed in the

manufacture of dairy products, certain foods, in manufacture of certain chemicals and in

numerous other ways.

Despite the established useful functions and potentially valuable activities of

microorganisms, these microscopic forms of life may be best known as agents of food spoilage

and causal agents of human beings viz. Acquired Immune Deficiency Syndrome (AIDS), herpes,

primitive and civilized societies have experienced diseases caused by microbes, frequently with

disastrous results. Moreover, microorganisms have played profound roles in warfare, religion

and the migration of populations.

Control of microbial population is necessary to prevent transmission of diseases,

infection, decomposition; contamination and spoilage caused by them.299 Man’s personal

comforts and convenience depend to a large extent on the control of microbial population.


BACTERIA:

In 1928, a German scientist C.E. Chrenberg first used the term ‘bacterium’ to denote

small microscopic organisms with a relatively simple and primitive form of the cellular

organization known as “prokaryotic”.

Danish physician, Gram, in 1884, discovered the famous Gram stain which categorisis bacteria

into two broad divisions: “Gram-positive” and “Gram-negative”. The Gram-positive bacteria

resist decolourization with acetone, alcohol and remain stained dark purple (Methyl Violet),

while Gram-negative bacteria are decolourized.300

According to morphological peculiarity, bacteria are generally unicellular e.g. cocci,

bacilli, etc., filamentous e.g. Actinomycetes, some being sheathed having certain cells

specialized for reproduction. The microorganisms capable of producing diseases in host are

known as ‘pathogenic’. Most of the microorganisms present on the skin and mucous membrane

are non-pathogenic and are often referred to as “commensals” or if they live on food residues as

in intestine, they may be called “saprophytes”. Generally, the pathogenic cocci and bacilli are

Gram-positive and the pathogenic coccobacilli are Gram-negative.

For evaluation of antibacterial activity in our case, we have used Staphylococcus aureus

and Streptococcus pyogenes from Gram-positive group of bacteria and Escherichia coli,

Pseudomonas aeruginosa, Salmonella enterica and Vibrio parahaemolyticus from Gram-

negative group of bacteria.

STAPHYLOCOCCUS AUREUS:

Genus: Staphylococcus [Microccaceae]

Staphylococci Are Differentiated From Micrococcus, A Genus Of The Same Family By Its

Ability To Utilize Glucose, Mannitol And Pyruvate Anaerobically. Cells Of Staphylococci Are

Usually To Be Found On The Skin Or Mucous Membranes Of The Animal Body, Especially Of

The Nose And Mouth Where They Occur In Large Numbers Even Under Normal Conditions.


Species: Staphylococcus Aureus

The Individual Cells Of S.Aureus Are 0.8 To 0.9 Micro In Diameter. They Are Ovoid Or

Spherical, Non Motile, Non Capsulated, Non Sporing Stain With Ordinary Aniline Dyes And

Gram Positive, Typically Arranged In Groups Of Irregular Clusters Like Branches Of Groups

Found In Pus, Singly Or In Pairs. The Optimum Temperature For The Growth Us 370 C,

Optimum Ph Is 7.4 To 7.6. They Produce Golden Yellow Pigment, Which Develops Best At

Room Temperature. They Cause Pyoregenic Of Pus Forming [Suppurative] Conditions, Mastitis

Of Women And Cows, Boils, Carbuncles Infantile Impetigo, Internal Abscess And Food

Poisoning.

ESCHERICHIA COLI :

Genus: Escherichia [Enterobacteriaceae]

This Genus Comprises Escherichia And Several Variants And Are Of Particular Interest

To The Sanitarian Since They Occur Commonly In The Formal Intestinal Tract Of Man And

Animals. Their Presence In Foods Or In Drinking Water May Indicates Faecal Pollution. E.Coli

Is The Most Distinctively Recognized Feacal Species.

Species: Escherichia coli

E.Coli is the most important type in this species, which contains a number of other

types.escherichia in 1885 discovered in from the faces of the newborn and showed the organisms

in the intesting within three days after birth. It is a Commensals of the human intesting and found

in the intestinal tract of men and animals and is also found in the sewage water, land, soil

contaminated by Feacal matters. The gram negative rods are 2 to 4 micro by 0.4 micro in size,

commonly seen in coccobacillary form and rarely in filamentous form. They are facultative

anaerobes and grow in all laboratory media. Colonies are circular, raised, smooth and emit a

faecal odour. E.Coli are generally non pathogenic and are incriminated as pathogens because in

certain instances some strains have been found to produce septicemia, inflammations of liver and


gall bladder, appendix, meningitis, pneumonia and other infections and this species is a

recognized pathogen in the veterinary field.

STREPTOCOCCUS PYOGENES :

Genus: Streptococcus

The term Streptococcus was first introduced by Bilroth [1874] and the term

Streptococcus Pyogenes was used by Rosenbach [1884]. These are spherical or ovoid cells;

divide in one axis and form chains; nonmotile and nonsporing. The growth is absence of native

proteins in the medium; they produce characteristic haemolytic changes in media containing

blood; produce acid only by fermentation of carbohydrates; often fail t liquefy gelatin; some

strains produce exotoxin and extracellular products; a few of them are Anaerobic.

Species: Streptococcus Pyogenes

Streptococcus Pyogenes is pathogenic to human and found in sore throat, follicular

tonsillitis, septicemia, acute or malignant ulcerative endocarditis etc. These are spherical Cocci

0.5 to 0.75 micro in diameter, arranged in moderately long chains of round Cocci and easily

differentiated from Enterococci that from short chains of 2 to 4 spheres. Streptococcus Pyogenes

is recently isolated from throat or other lesions; they show either mucoid or matt colonies. On

keeping in the laboratory, they undergo varation to a glossy type. Streptococci are susceptible to

destructive agents, and to penicillin and sulphomamides.

PSEUDOMONAS AERUGINOSA :

Genus: Pseudomonas

Genus: Pseudomonas is characterized by gram negative motile rods, nonsporing aerobes,

oxidase positive, bluish green or yellowish pigment diffusing into the medium. Out of 140

species, only one is pathogenic to human.

Species: Pseudomonas aeruginosa


Ps.aeruginosa occurs as a commensal in the intestine of human and animal’s but, when

the defensive mechanism of the body is poor. It acts as a minor pathogen producing Suppurative

wound, otitis media, peritonitis, cystitis, bronchopneumonia and empyema. In children it causes

diarrhea and septicemia. The pus produced by p.aeruginosa is greenish blue. These are gram

negative, actively motile, non sporing organisms 1.5-3 micro by 0.5 micro with rounded ends

and bipolar flagella. They occur singly or in pair, of short chains. They grow well in ordinary

media under aerobic conditions, producing diffusible pigment.

SALMONELLA ENTERICA :

Genus: Salmonella

Salmonella enterica is a rod shaped, flagellated, Gram-negative bacterium, and a member

of the genus Salmonella. The bacteria is characterized by its flagellar antigen, H, its

lipopolysaccharidic O antigen, and, in addition, it’s PS capsular Vi (for virulence) antigen, found

at the surface of freshly isolated strains.

Species: S. typhi

S. enterica has an extraordinarily large number of serovars or strains-up to 2000 have

been described. Salmonella enterica Serovar Typhi (historically elevated to species status as S.

typhi) is the disease agent in typhoid fever. Other serovars such as Typhimurium (also known as

S. typhimurium) can lead to a form of human gastroenteritis sometimes referred to as

salmonellosis. Salmonella Typhi is a serovar of Salmonella enterica (formerly known as

Salmonella choleraesuis) and the cause of the disease typhoid fever. The organism can be

transmitted by the fecal-oral route-it is excreted by humans in feces and may be transmitted by

contaminated water, food, or by person-to-person contact.

VIBRIO PARAHAEMOLYTICUS :

Genus: Vibrio


Vibrio parahaemolyticus is a curved, rod-shaped, Gram-negative bacterium found in

brackish saltwater, which, when ingested, causes gastroeintestinal illness in humans. V.

parahaemolyticus is oxidase positive, facultatively aerobic, and does not form spores.

Species: Motile

While infection can occur via the fecal-oral route, ingestion of bacteria in raw or

undercooked seafood, usually oysters, is the predominant cause the acute gastroenteritis caused

by V. parahaemolyticus. Wound infections also occur, but are less common than seafood-borne

disease. The disease mechanism of V. parahaemolyticus infections has not been fully elucidated.

However, most clinical disease results from strains that carry either the thermostable direct

hemolysin gene (tdh) or the tdh-related hemolysin gene (trh) or both genes.

FUNGI

Introduction:

It has been estimated that the life span of humans has increased by almost a decade since

the discovery of antimicrobial agents against microbial infections. A consequence of our success

with antibacterial agents and improved medical care is the increase in the number of fungal

infections.

The incidence of fungal infection has increased dramatically in the past 20 years partly

due to increase in the number of people whose immune systems are compromised by AIDS,

aging, organ transplantation or cancer therapy. Accordingly, the increase in rates of morbidity

and mortality of infections has been now recognized as a major problem. In response,

pharmaceutical industry has developed a number of novel less toxic antifungals for clinical use.

Its increased use often for prolonged period, has led to the increased incidence of infections with

less common species, with intrinsic resistance to one or more of the available antifungal agents.

Fungi are non photosynthetic eukaryotes growing either as a colony of single cells

(yeasts) or as filamentous multicellular aggregates (molds). Most fungi live as saprobes in soil or

dead plant material and are important in the mineralization of organic matter. Some are


pathogens of humans and animals. The in vitro methods used for detection of antifungal potency

are similar to those used in antibacterial screening. As with bacteria, it is easy to discover several

synthetic and natural compounds that, in minute quantities, can retard or prevent growth of

fungi.

CANDIDA ALBICANS:

Genus: Candida

Candida species reproduce by yeast like budding cells but they also show formation of

pseudomycellum. These pseudomycellum are chains of elongated cells formed from buds and

the buds elongated without breaking of the mother cell. They are very fragile and separate easily.

Mycelia also form by the elongation of the germ tube produced by a mother cell.

Species: Candida albicans

Candida albicans may remain as a commensal of the mucous membrane with of without

causing any pathologic changes to the deeper tissues of the same fungus may cause pathological

lesion of the skin. Such a fungus under favorable conditions can cause superficial, intermediate

of deep mycoses depending on the condition of the host.

ASPERGILLUS NIGER AND ASPERGILLUS CLAVATUS:

Genus: Aspergillus

The Aspergilli are widespread in nature, being found on fruits, vegetables and other

substrates, which may provide nutriment. Some species are involved in food spoilage. They are

important economically because they are used in a number of industrial fermentations, including

the production of citric acid gluconic acid. Aspergilli grow in high concentrations of sugar and

salt, indicating that they can extract water required for their growth from relatively dry

substances.


EVALUTION TECHNIQUE

1. The following conditions must be met for the screening of antimicrobial activity:

(a) There should be intimate contact between the test organisms and substance to be

evaluated.

(b) Required conditions should ne provided for the growth of microorganisms.

(c) Conditions should be same through the study.

(d) Aseptic / sterile environment should be maintained.

Various methods have been used from time to time by several workers to evaluate the

antimicrobial activity. The evaluation can be done by the following methods301-303:

i. Turbidometric method.

ii. Agar streak dilution method.

iii. Serial dilution method.

iv. Agar diffusion method.

Following Techniques are used as agar diffusion method:

i. Agar Cup method.

ii. Agar Ditch method.

iii. Paper Disc method.

Broth dilution method, a widely used non-automated in vitro bacterial susceptibility test has

been used to evaluate antimicrobial activity.

It is a classic method that yields a quantitative result for the amount of antimicrobial

agent that is needed to inhibit growth of microorganisms. It is carried out in tubes.

i. Macro dilution method in tubes.

ii. Micro dilution format using plastic trays.

iii. In the present protocol we have used the Micro dilution format.

iv. Determination of Minimal Bactericidal Concentrations (MBC) by Broth Dilution

Method:

Materials and method:


1. All the synthesized drugs were used for antibacterial tests.

2. All necessary controls, viz. drug, vehicle, broth of organism were used.

3. The drug Gentamycin was used as control.

4. Muller Hinton Broth was used as nutrient medium to grow the strains and

dilute the drug suspensions for

5 Test. all MTCC cultures were tested against above-mentioned known and unknown

Drugs.

6. Serial dilution technique was followed by micro method as per NCCLS-1992 manual.304

7. Inoculum size: Inoculum size for test strain was adjusted to 108 cfu (colony forming unit)

per ml.

8. The strains used for screening of antibacterial and antifungal activities were, the strains

procured from Institute of Microbial Technology (IMTECH), Chandiharh.

The following stains procured from IMTECH-Chandigarh were used for

screening antibacterial and antifungal activities.

i. Escherichia coli [Gram negative] MTCC – 442

ii. Pseudomonas aeruginosa [Gram negative] MTCC – 441

iii. Staphylococcus aureus [Gram positive] MTCC – 96

iv. Streptococcus Pyogenes [Gram positive] MTCC – 443

v. Salmonella typhi [Gram negative] MTCC – 98

vi. Vibrio haemolyticus [Gram negative] MTCC – 451

vii. Candida albicans Fungus] MTCC – 227

viii. Aspergillus Niger [Fungus] MTCC – 282

ix. Aspergillus clavatus Fungus] MTCC – 1323

9. DMSO was used as diluent/vehicle to get desired concentration of drugs to test upon

standard bacterial strains.

Minimal Bactericidal Concentrations (MBC)


The main advantage of the ‘Broth Dilution Method’ for MBC determination lies in the fact that

it can readily be converted to determined the MBC as well.

1) Serial dilutions were prepared in primary and secondary screening.

2) The control tube containing no antibiotic is immediately subcultured (before incubation)

by spreading a loopful evenly over a quarter of plate of medium suitable for the growth

of the test organism and incubated at 370C for 24 h.

3) The MBC of the control organism is read to check the accuracy of drug concentrations.

4) The lowest concentration that inhibits growth of the organism is recorded as the MBC.

5) All the tubes not showing visible growth (in the same manner as control tube described

above) are subcultured and incubated overnight at 370C.

6) The amount of growth from the control tube before incubation (which represents the

original inoculum) was compared.

7) Subcultures may show similar number of colonies indicating bacteriostatic,a reduced

number of colonies – indicating a partial or slow bactericidal activity and no growth – if

the whole inoculum has been killed. The test must include a second set of the same

dilutions inoculated with an organism of known sensitivity.

Minimal fungicidal concentration (MFC)

Broth Dilution Method for MFC Determation lies in the fact that it can readily be

converted to determine the MFC as well.Serial dilution where primery and secondary screening

and the material and method was just followed like a bacterialcidal activity. The growth,

inhibition is masured and compound is applied in the method to determine the activity in µg/ml

concentration.

Methods used for primary & secondary screening:

Each synthesized drug was diluted obtaining 2000 microgram /ml concentration, as a stock

solution.


Primary screen: In primary screening 500 micro/ml, 250 micro/ml, and 125

micro/ml concentrations of the synthesized drugs were taken. The active synthesized

drugs found in this primary screening were further tested in a second set of dilution

against all microorganisms.

Secondary screen: The drugs found active in primary screening were similarly

diluted to obtain 100 micro/ml, 50 micro/ml, 25 micro/ml, 12.5 micro/ml, 6.250

micro/ml, 3.125 micro/ml and 1.5625 micro/ml concentrations.

Reading Result:-

The highest dilution showing at least 99 % inhibition zone is taken as MIC.The

result of this is much affected by the size of the inoculum. The test mixture should

contain 108 organism/ml.

The standard drug:

The standard drug used in the present study is “Gentamycin” for evaluating antibacterial activity

which showed (0.25, 0.05, 0.5 & 1 μg/ml). MBC against S. aureus, E. pyogenes & P. aeruginosa

respectively. “K.Nystatin” is used as the standard drug for antifungal activity which showed 100

μg/ml MFC against all the species, used for the antifungal activity.


IMPORTANT ANTIBACTERIAL AND ANTIFUNGAL RUGS

OO

O NH2

MeNH2

NH2

OH

NH2

OH

OH

Me

NH

Me

Gentamicin

NH2

NH

O N

S

O

OHO

H

OH

NH

OH

Cl

Cl

O

O2N

Ampicillin Chloramphenicol

NN

NH

OH

OO

F

NN

NH

OH

OO

F

Ciprofloxacin Norfloxacin


CH3

O

OHO OH OH OH O OH

OH

CH3

CH3

HO

OH

OO

OH NH2

OH

CH3

COOH

K.

Nystatin

Experimentation carried out for antibacterial screening


Experimentation carried out for antibacterial screening


Graphilcal Chart No. 1: Antimicrobial activity of 2-(Adamantan-1-yl-{4-[4-(Aryl-amino)-phenyl]-thiazol-2-yl}-amino)-acetamide

(ASI-01-10)

0

5

10

15

20

25

30

A B C D F

ZONE OF INHIBITION IN mm

P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin

C=Tetracyclin D=Gentamycin

D=Chloramphenicol

ASI‐01 ASI‐02 ASI‐03 ASI‐04 ASI‐05 ASI‐06 ASI‐07 ASI‐08 ASI‐09 ASI‐10


Graphilcal Chart No. 2: Antimicrobial activity of 2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-Aryl-acetamide (ASII-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASII‐01 ASII‐02 ASII‐03 ASII‐04 ASII‐05 ASII‐06 ASII‐07 ASII‐08 ASII‐09 ASII‐10


Graphilcal Chart No. 3: Antimicrobial activity of 2-(Adamantan-1-yl-{4-[4-(ARYL-amino)-phenyl]-thiazol-2-yl}-amino)-N-propyl-acetamide (ASIII-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASIII‐01 ASIII‐02 ASIII‐03 ASIII‐04 ASIII‐05 ASIII‐06 ASIII‐07 ASIII‐08 ASIII‐09 ASIII‐10


Graphilcal Chart No. 4: Antimicrobial activity 2-(Adamantan-1-yl-{4-[4-(4-oxo-2-ARYL-thiazolidin-3-yl)-phenyl]-thiazol-2-yl}-amino)-N-phenyl-acetamide (ASIV-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASIV‐01 ASIV‐02 ASIV‐03 ASIV‐04 ASIV‐05 ASIV‐06 ASIV‐07 ASIV‐08 ASIV‐09 ASIV‐10


Graphilcal Chart No. 5: Antimicrobial activity 1-(Sec-amine)-4-YL-2-[5-PHENYL-3-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-

YL)-[1,2,4]TRIAZOL-1-YL]-ETHANONE (ASV-01-06)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASV‐01 ASV‐02 ASV‐03 ASV‐04 ASV‐05 ASV‐06


Graphilcal Chart No. 6: Antimicrobial activity of N-(Aryl)-2-[5-PHENYL-3-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-YL)-

[1,2,4]TRIAZOL-1-YL]-ACETAMIDE (ASVI-01-08)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASVI‐01 ASVI‐02 ASVI‐03 ASVI‐04 ASVI‐05 ASVI‐06 ASVI‐07 ASVI‐08


Graphilcal Chart No. 7: Antimicrobial activity of Aryl-{4-[2-ethyl-5-(1-phenyl-3,4-dihydro-1H-isoquinolin-2-yl)-2H-[1,2,4]triazol-3-yl]-phenyl}-amine (ASVII-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASVII‐01 ASVII‐02 ASVII‐03 ASVII‐04 ASVII‐05 ASVII‐06 ASVII‐07 ASVII‐08 ASVII‐09 ASVII‐10


Graphilcal Chart No. 8: Antimicrobial activity of N-{4-[2-ETHYL-5-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-YL)-2H-

[1,2,4]TRIAZOL-3-YL]-ARYL}-BENZAMIDE (ASVIII-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASVIII‐01 ASVIII‐02 ASVIII‐03 ASVIII‐04 ASVIII‐05 ASVIII‐06 ASVIII‐07 ASVIII‐08 ASVIII‐09 ASVIII‐10


Graphilcal Chart No. 9: Antimicrobial activity of N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-ARYL/ALKYLamide

(ASIX-01-10)

0

5

10

15

20

25

30

A B C D F


P. aeruginosa

S.Aureus

E.Coli S. pyogenes

A= DMSO B=Ampicilin


D=Chloramphenicol

ASIX‐01 ASIX‐02 ASIX‐03 ASIX‐04 ASIX‐05 ASIX‐06 ASIX‐07 ASIX‐08 ASIX‐09 ASIX‐10


TABLE: 13

ANTIBACTERIAL ACTIVITY OF

2-(Adamantan-1-yl-{4-[4-(Aryl-amino)-phenyl]-thiazol-2-yl}-amino)-acetamide

OF THE FOLLOWING TYPE

Sr.No.

-Ar

MINIMAL BACTERICIDAL CONCENTRATIONS

(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml

S. pyogenes MTCC 442

µg / ml

S.Typhi TCC 98 µg / ml

V.Parahaemolyticus

MTCC 451 µg / ml

ASI-01 -C6H5 250 250 100 500 1000 500

ASI-02 -C6H4-CH3 1000 1000 125 500 500 250

ASI-03 -C6H4-Br 200 250 125 500 200 500

ASI-04 -C6H4-CN 125 500 125 500 1000 500

ASI-05 C6H4COCH3 1000 1000 500 500 125 125

ASI-06 -CH2-C6H5 500 1000 1000 500 1000 500

ASI-07 -C6H4-C4H9 1000 500 500 1000 500 500

ASI-08 -C6H4-C3H7 500 500 250 250 500 1000

ASI-09 -C5H4N 500 250 500 250 1000 500

ASI-10 -C6H3-F2 1000 500 250 500 100 1000

Antibacterial activity results showed that Compound -C6H4-CH3 and C6H4-Br was

moderately active against S. aureus. Compound -C6H4-CN was moderately active against S.

aureus and E. coli. Compound C6H4COCH3 was moderately active against both gram

positive bacteria S.Typhi and V.Parahaemolyticus. Compound was -C6H3-F2 moderately

active against S. aureus.


TABLE: 14


2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-Aryl-acetamide


Sr.No.

-Ar


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml


µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASII-01 -C6H5 250 500 1000 500 250 500

ASII-02 -C6H4-CH3 500 250 500 500 250 1000

ASII-03 -C6H4-Br 250 250 1000 250 500 250

ASII-04 -C6H4-CN 200 500 1000 200 500 1000

ASII-05 C6H4COCH3 500 250 500 250 250 500

ASII-06 -CH2-C6H5 250 200 250 500 500 250

ASII-07 -C6H4-C4H9 100 250 125 200 500 1000

ASII-08 -C6H4-C3H7 250 50 100 125 1000 500

ASII-09 -C5H4N 250 500 500 250 500 250

ASII-10 -C6H3-F2 200 250 100 125 500 250

Antibacterial activity results showed that compound -C6H4-C4H9 was moderately

active against E. coli and S. aureus. Compound -C6H4-C3H7 was quite well active against P.

aeruginasa and moderately active against S. aureus. and -C6H3-F2 was moderately active

against E. coli and S. aureus.and S. pyogenes

.


TABLE: 15


2-(Adamantan-1-yl-{4-[4-(ARYL-amino)-phenyl]-thiazol-2-yl}-amino)-N-propyl-

acetamide


Sr.No.

-Ar


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml

S. pyogenes MTCC

442

µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASIII-01 -C6H5 100 62.5 250 200 100 500

ASIII-02 -C6H4-CH3 200 250 250 250 500 500

ASIII-03 -C6H4-Br 62.5 100 250 500 500 250

ASIII-04 -C6H4-CN 125 125 62.5 500 500 500

ASIII-05 C6H4COCH3 100 62.5 200 250 500 200

ASIII-06 -CH2-C6H5 252 125 250 125 500 500

ASIII-07 -C6H4-C4H9 500 1000 250 500 1000 500

ASIII-08 -C6H4-C3H7 1000 1000 500 250 50 100

ASIII-09 -C5H4N 500 250 1000 250 1000 500

ASIII-10 -C6H3-F2 1000 500 500 1000 100 100

Antibacterial activity results showed that, compound -C6H5 was quite well active against P.

aeruginosa and moderately active against E. coli and andS.Typhi-Compound -C6H4-Br was

quit well active against E. coli and moderately active against P. aeruginosa. C6H4-

COCH3C was active against both gram negative bacteria E. coli and P. aeruginosa.

Compound -C6H4-C3H7 was active against S.Typhiand and V.Parahaemolyticus.

Compound -C6H4-CN was active against S aureus.Compound -C6H3-F2 was moderately

active against S.Typhiand and V.Parahaemolyticus.


TABLE: 16


2-(Adamantan-1-yl-{4-[4-(4-oxo-2-ARYL-thiazolidin-3-yl)-phenyl]-thiazol-2-yl}-

amino)-N-phenyl-acetamide


Sr.No.

-Ar


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml


µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASIV-01 -C6H5 62.5 100 200 250 100 500

ASIV-02 -C6H4-CH3 200 125 100 500 250 500

ASIV-03 -C6H4-Br 62.5 62.5 100 62.5 500 125

ASIV-04 -C6H4-CN 100 50 62.5 250 250 500

ASIV-05 C6H4COCH3 125 125 100 500 500 200

ASIV-06 -CH2-C6H5 500 125 100 200 500 200

ASIV-07 -C6H4-C4H9 200 250 125 500 250 500

ASIV-08 -C6H4-C3H7 500 1000 1000 125 50 200

ASIV-09 -C5H4N 125 250 200 125 1000 500

ASIV-10 -C6H3-F2 500 100 125 200 500 1000

Antibacterial activity results showed that compound -C6H5 was active against E. coli and

moderately active against S. aureus and S.Typhi . Compound -CH2-C6H5,-C6H4COCH3,-C6H4-

CH3 and -CH2-C6H5 was moderately active against S. aureus. Compound -C6H4-Br more

active against E. coli, S. pyogenes and P. aeruginasa moderately active against S. aureus.

Compound -C6H4-CN was active against P. aeruginosa and S. aureus. Compound -C6H4-

C3H7 was active against E. coli S.Typhi. Compound -C6H3-F2 moderately active against P.

aeruginasa.


TABLE: 17


1-(Sec-amine)-4-YL-2-[5-PHENYL-3-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-

YL)-[1,2,4]TRIAZOL-1-YL]-ETHANONE


Sr.No.

-R


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aerugino

sa

MTCC 1688

/ l

S. aureus

MTCC 96

/ l

S. pyogenes MTCC

442

µg / ml

S.Typhi TCC

98 µg / ml

V.Parahaemolyticu

s MTCCASV-01 -NC4H8O 200 62.5 200 250 500 200

ASV-02 -NC5H10 62.5 100 62.5 500 250 1000

ASV-03 -NC4H8 62.5 125 100 500 1000 1000

ASV-04 -NC4H8NCH3 500 250 500 200 1000 500

ASV-05 -NC4H8NC6H5 125 100 50 125 500 500

ASV-06 -NC5H9C6H5 1000 500 500 1000 500 500

Antibacterial activity results showed that compound -NC4H8O was good active against

Gram negative bacteria, P. aeruginosa. Compound -NC5H10 was good active against E. coli

and S aureus, while moderately active against P. aeruginosa. Compound -NC4H8 good

active against bacteria E. coli and moderately active against S. aureus. Compound -

NC4H8NC6H5 was more active against S aureus and moderately active against P.

aeruginosa


TABLE: 18





Sr.No.

-Ar


(MBC) IN µg / ml

E. coli

MTCC 443 µg /

ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml

S. pyogenes MTCC

442

µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASVI-01 -C6H5 500 250 200 500 100 250

ASVI-02 -C6H4CH3 250 500 500 250 500 200

ASVI-03 -C6H4Br 500 100 500 500 250 250

ASVI-04 -C6H4CN 250 200 250 500 250 500

ASVI-05 C6H4COCH3 500 250 250 500 250 250

ASVI-06 -CH2C6H5 500 500 500 500 500 1000

ASVI-07 -C6H4C4H9 200 500 1000 200 100 500

ASVI-08 -C6H4C3H7 250 500 1000 125 200 1000

Antibacterial activity results showed that compound -C6H5 and -C6H4C4H9 moderat activity

against S.Typhi. and compound was -C6H4Br moderat activity against P. aeruginosa.Rest

of the compounds did not show satisfactory activity against these species.


TABLE: 19


N-{4-[2-ETHYL-5-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-YL)-2H-

[1,2,4]TRIAZOL-3-YL]-ARYL}-BENZAMIDE


Sr.No.

-Ar


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml

S. pyogenes MTCC

442

µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASVIII-01 -C6H5 500 200 500 200 250 500

ASVIII-02 -C6H4CH3 200 250 500 125 500 500

ASVIII-03 -C6H4Br 250 500 200 200 500 500

ASVIII-04 -C6H4CN 200 500 200 250 500 100

ASVIII-05 -C6H4F 125 250 250 500 250 250

ASVIII-06 -C6H4Cl 200 250 500 250 200 500

ASVIII-07 -C6H4NO2 125 200 250 125 500 500

ASVIII-08 -C5H4N 250 250 250 62.5 250 1000

ASVIII-09 -CH2C6H5 200 125 125 50 100 500

ASVIII-10 -C6H4OH 500 250 200 250 500 250

Antibacterial activity results showed that compound -C5H4N was shawing good

activity against S. Pyogenes. Compound -CH2C6H5 was more active against S. Pyogenes

and moderately active against S.Typhi.


TABLE: 20


N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-ARYL/ALKYL

AMIDE


Sr.No.

-R


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml


µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASIX-01 -C6H5 500 62.5 200 250 250 100

ASIX -02 -C6H4-CH3 200 100 125 500 250 500

ASIX -03 -C6H4-Br 200 250 500 125 500 250

ASIX 04 -C6H4-CN 250 200 125 250 500 1000

ASIX -05 -CH2-C6H5 125 200 100 100 250 500

ASIX -06 -C6H4-C4H9 200 250 200 250 200 500

ASIX -07 -C6H4-C3H7 500 62.5 62.5 500 500 1000

The data of antibacterial activity of this series indicated that compound -C6H5 was having

good activity against P. aeruginosaand -C6H4CH3 was moderately active. Compound -CH2-

C6H5 was moderately active against S. aureusand S. pyogenes.. Compound -C6H4-C3H7 was

shaving good activity against both gram negative bacteria E. coli and P. aeruginasa.


TABLE: 21


N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-aryl/alkyl-

SULFONAMIDE


Sr.No.

-R


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96 µg /

ml


µg / ml


V.Parahaemolyticus

MTCC 451 µg / ml

ASX-01 -CH3 62.5 100 62.5 100 200 250

ASX -02 -C2H5 125 250 50 250 500 1000

ASX -03 -CH(CH3)2 250 200 200 200 250 500

ASX -04 -C6H5 50 200 50 250 200 500

ASX -05 -C6H4-CH3 200 250 125 200 250 500

ASX -06 -C6H4-Br 62.5 250 50 200 500 250

ASX -07 -C6H4-Cl 200 250 125 250 200 500

ASX -08 -C6H4-F 100 500 62.5 200 250 500

ASX -09 -C6H4C3H7 100 50 25 500 500 500

ASX -10 -C6H4-NO2 200 250 125 250 200 500

Antibacterial activity results showed that compound -CH3 was shawing good activity

against all species. Compound -C2H5 was active against S. Aereus. Compound -C3H7 was

moderately active against E. coli and more active against S. aureus. Compound -C6H4-Br

was more active against E. coli and S. aureus. Compound -C6H4C3H7 was most active

against S. aureus and more active against both gram negative bacteria E. coli and P.

aeruginosa.


TABLE: 22


THE STANDARD DRUGS

Sr.No.


(MBC) IN µg / ml

E. coli

MTCC 443

µg / ml

P. aeruginosa

MTCC 1688

µg / ml

S. aureus

MTCC 96

µg / ml


µg / ml

GENTAMYCIN 0.05 1 0.25 0.5

AMPICILLIN 100 100 250 100

CHLORAMPHENICOL 50 50 50 50

CIPROFLOXACIN 25 25 50 50

NORFLOXACIN 10 10 10 10


TABLE: 23

ANTIFUNGAL ACTIVITY OF

2-(Adamantan-1-yl-{4-[4-(Aryl-amino)-phenyl]-thiazol-2-yl}-amino)-acetamide


Sr.No.

-Ar

MINIMAL FUNGICIDAL CONCENTRATIONS

(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASI-01 -C6H5 1000 500 500

ASI-02 -C6H4-CH3 500 1000 1000

ASI-03 -C6H4-Br 500 1000 500

ASI-04 -C6H4-CN 1000 500 1000

ASI-05 C6H4COCH3 500 1000 500

ASI-06 -CH2-C6H5 >1000 500 1000

ASI-07 -C6H4-C4H9 500 1000 1000

ASI-08 -C6H4-C3H7 >1000 500 500

ASI-09 -C5H4N 500 1000 1000

ASI-10 -C6H3-F2 >1000 >1000 >1000

The data of antifungal activity of this series indicated that, compounds were not showing

potential activity again any species.


TABLE: 24


2-{Adamantan-1-yl-[4-(4-amino-phenyl)-thiazol-2-yl]-amino}-N-Aryl-acetamide


Sr.No.

-Ar


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASII-01 -C6H5 500 1000 >1000

ASII-02 -C6H4-CH3 1000 >1000 500

ASII-03 -C6H4-Br 1000 1000 >1000

ASII-04 -C6H4-CN 1000 500 500

ASII-05 C6H4COCH3 >1000 1000 500

ASII-06 -CH2-C6H5 1000 500 1000

ASII-07 -C6H4-C4H9 1000 500 >1000

ASII-08 -C6H4-C3H7 500 >1000 1000

ASII-09 -C5H4N 1000 1000 500

ASII-10 -C6H3-F2 1000 1000 1000




TABLE: 25


2-(Adamantan-1-yl-{4-[4-(ARYL-amino)-phenyl]-thiazol-2-yl}-amino)-N-propyl-acetamide


Sr.No.

-Ar


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASIII-01 -C6H5 1000 500 500

ASIII-02 -C6H4-CH3 1000 1000 1000

ASIII-03 -C6H4-Br 500 500 500

ASIII-04 -C6H4-CN >1000 1000 >1000

ASIII-05 C6H4COCH3 500 >1000 1000

ASIII-06 -CH2-C6H5 500 500 1000

ASIII-07 -C6H4-C4H9 500 >1000 500

ASIII-08 -C6H4-C3H7 >1000 1000 >1000

ASIII-09 -C5H4N 1000 500 500

ASIII-10 -C6H3-F2 1000 1000 1000




TABLE: 26





Sr.No.

-Ar


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASVI-01 -C6H5 1000 1000 1000

ASVI-02 -C6H4CH3 500 >1000 >1000

ASVI-03 -C6H4Br >1000 1000 1000

ASVI-04 -C6H4CN 500 500 >1000

ASVI-05 C6H4COCH3 1000 1000 1000

ASVI-06 -CH2C6H5 500 200 500

ASVI-07 -C6H4C4H9 1000 1000 1000

ASVI-08 -C6H4C3H7 200 200 200

The data of antifungal activity of this series indicated that, compound -C6H4C3H7 was

active against C. albicans,, A. niger and A. clavatus and -CH2C6H5 was active against A. niger.

Rest of compounds was not showing potential activity again any species.


TABLE: 27


N-{4-[2-ETHYL-5-(1-PHENYL-3,4-DIHYDRO-1H-ISOQUINOLIN-2-YL)-2H-

[1,2,4]TRIAZOL-3-YL]-ARYL}-BENZAMIDE


Sr.No.

-Ar


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASVIII-01 -C6H5 500 500 1000

ASVIII-02 -C6H4CH3 >1000 1000 1000

ASVIII-03 -C6H4Br 1000 1000 >1000

ASVIII-04 -C6H4CN 500 >1000 500

ASVIII-05 -C6H4F >1000 1000 1000

ASVIII-06 -C6H4Cl 1000 500 >1000

ASVIII-07 -C6H4NO2 500 1000 500

ASVIII-08 -C5H4N >1000 500 1000

ASVIII-09 -CH2C6H5 1000 500 >1000

ASVIII-10 -C6H4OH 500 1000 1000




TABLE: 28


N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-aryl/alkyl-

SULFONAMIDE


Sr.No.

-R


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASX-01 -CH3 100 1000 500

ASX -02 -C2H5 1000 500 500

ASX -03 -CH(CH3)2 500 500 1000

ASX -04 -C6H5 500 1000 1000

ASX -05 -C6H4-CH3 500 1000 1000

ASX -06 -C6H4-Br 1000 500 1000

ASX -07 -C6H4-Cl 500 1000 500

ASX -08 -C6H4-F 1000 500 1000

ASX -09 -C6H4C3H7 1000 500 1000

ASX -10 -C6H4-NO2 500 1000 500

The data of antifungal activity of this series indicated that, compound -CH3 was exhibit

very good activity against C. albicans, Rest of compounds was not showing potential activity

again any species.


TABLE: 29


N-(1-CYANO-1,4-DIHYDRO-CYCLOPENTA[C]CHROMEN-2-YL)-ARYL/ALKYL AMIDE


Sr.No.

-Ar


(FBC) in μg/ml

C. albicans

MTCC 227

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

ASIX-01 -C6H5 1000 >1000 >1000

ASIX -02 -C6H4-CH3 500 500 500

ASIX -03 -C6H4-Br 1000 1000 1000

ASIX 04 -C6H4-CN 500 100 500

ASIX -05 -CH2-C6H5 1000 500 1000

ASIX -06 -C6H4-C4H9 1000 1000 100

ASIX -07 -C6H4-C3H7 200 500 1000

The data of antifungal activity of this series indicated that, compound -C6H4-C3H7 was

exhibit good activity against C. albicans, Rest of compounds was not showing potential activity

again any species.


TABLE: 30


THE STANDARD DRUGS

Sr.No.


(FBC) in μg/ml

C. albicans

MTCC 227

μg/ml

A. niger

MTCC 282 μg/ml

A. clavatus

MTCC 1323

μg/ml

NYSTATIN 100 100 100

GRESEOFULVIN 500 100 100

chapter iv result and discussion -...

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