experimental work - shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/7005/20/10_chapter...

Chapter 4

School of Pharmacy & Technology Management, SVKM’s NMIMS, Mumbai

The section consists of following subsections –

A. Preparation of library of synthesizable compounds and structure based design of 1,3-

diarylpropenone analogues

B. Synthesis of designed 1,3-diarylpropenone analogues

C. Profiles of reactants and synthesized compounds

D. Pharmacological evaluation of synthesized analogues

E. Quantitative Structure Activity Relationship (QSAR) study of synthesized molecules

based on pharmacological evaluation

EXPERIMENTAL

WORK

CHAPTER- 4 EXPERIMENTAL

School of Pharmacy & Technology Management, SVKM’s NMIMS, Mumbai 23

4 EXPERIMENTAL

A. PREPARATION OF SET OF SYNTHESIZABLE COMPOUNDS AND

STRUCTURE BASED DESIGN OF 1,3-DIARYLPROPENONE ANALOGUES

Computational study was carried out to design 1,3-diarylpropenone analogues as VEGFR-2

inhibitors. In the study, binding of 1,3-diarylpropenone analogues with VEGFR-2 was

studied using computer aided molecular modeling techniques.

4.1 Softwares used in the study

• Molecular modeling softwares used for the study were -

o Schrödinger software [Maestro®

version 7.5 (Graphical user interface), USA] on Red

Hat Linux Enterprise platform (User Manual and Tutorial, 2006).

� Ligprep module

� Sitemap module

� Glide module

� QikProp module

� Protein preparation wizard

o CS ChemDraw Ultra version 7.01, Cambridge Soft Corporation USA

• The 3-dimensional (3D) structures of VEGFR-2 and 1,3-diarylpropenone analogues were

used for molecular docking study.

• The chemical structures were sketched in ChemDraw Ultra version 7.01, and saved in

MDL mol file format.

• The starting coordinates of crystal structures of VEGFR-2 were obtained from Protein

Data Bank, PDB (http://www.pdb.org/pdb/results/results.do?outformat=&qrid=

BCF8BFE2&tabtoshow=Current) and further modified for Glide docking calculations.

• For the docking of ligands into protein active sites and to estimate the binding affinities

of docked compounds, an advanced molecular docking program GLIDE, version 2.5

(Schrödinger Inc, USA), was used in this study (User Manuals 2006).

4.2 Virtual screening of 1,3-diarylpropenone analogues

The virtual screening of 1,3-diarylpropenone analogues was carried out using following

steps –

4.2.1 Evaluation of all 3D structures of VEGFR-2

4.2.2 Protein preparation of all VEGFR-2 structures



4.2.3 Study of known VEGFR-2 and angiogenesis inhibitors

4.2.4 Study of binding site of VEGFR-2

4.2.5 Design of test molecules

4.2.6 Ligand Preparation of designed 1,3-diarylpropenone analogues

4.2.7 Enrichment studies

4.2.8 Validation of docking protocol

4.2.9 Docking study using Glide

4.2.1 Evaluation of all 3D structures of VEGFR-2

All nineteen PDB structures (available at the time of study) of VEGFR-2 were chosen for the

study. Various programs like Procheck, What_check, Verify_3D and Errat using National

Institute of Health (NIH) server available online (http://nihserver.mbi.ucla.edu/SAVES/)

were used to validate the structures of the proteins. Full geometric analysis as well as stereo

chemical quality of the protein structure was performed by Procheck by analyzing residue-

by-residue geometry and overall structure geometry. Ramachandran plot statistics were used

to evaluate the stability of the model.

4.2.2 Protein Preparation of all VEGFR-2 structures

X-ray complexes of VEGFR-2 were imported to Maestro®

and the co-crystallized ligands

were identified.

All the crystallographic water molecules were deleted in initial docking experiment. The

water molecules forming bridge interactions between the protein residues and ligand atoms

were considered for the study. Bond orders for crystal ligand and protein were adjusted.

Once aligned, hydrogen atoms were added to all the protein–ligand complexes.

The guanidines and ammonium groups in all the arginine and lysine side chains were made

cationic and the carboxylates of aspartate and glutamate residues were made anionic.

Asparagine (Asn) side-chain amides were flipped by 180° to optimize the interactions with

the X-ray ligands. In addition, the Aspartic acid (Asp) side-chain carboxylate was treated in a

neutral form in the protein structures of all the complexes. The most likely positions of

hydroxyl and thiol hydrogen atoms, protonation states and tautomers of histidine (His)

residues, and Chi ‘flip’ assignments for Asn, Glutamine (Gln) and His residues were selected

by the protein assignment script.

Following the above steps of preparation, the protein ligand complexes in the X-ray

structures were subjected to energy minimization using the Schrödinger with implementation



of OPLS-2001 force field in the protein preparation wizard and refinement with implicit

solvation in two stages.

In the first stage, the positions of water molecules were optimized keeping the ligand and the

protein atoms in their X-ray structure positions. In the second stage, the entire complex was

minimized and minimization terminated when the root mean square deviation (RMSD) of

the heavy atoms in the minimized structure relative to the X-ray structure exceeded 0.3 A°.

This helps maintaining the integrity of the prepared structures relative to the corresponding

experimental structures, while eliminating bad contacts between heavy atoms.

4.2.3 Study of known VEGFR-2 and angiogenesis inhibitors

The known VEGFR-2 inhibitors present in market and clinical trials (Table 4.1) and

patented angiogenesis inhibitors (Table 4.2) were studied.

Table 4.1: List of known VEGFR-2 inhibitors (Molecules in market and clinical trials) (Selleck,

2010)

Sr.

No.

Compound Structure and IUPAC Name MW Mol

formula

IC50*

1. PTK787

Vatalanib

Cl

HN

NN

N

N-(4-chlorophenyl)-4-(pyridin-4-

ylmethyl)phthalazin-1-amine

346.81 C20H15ClN4 42 nM

2. SU5402

HN

O HN

O

OHH

3-(2-{[(3Z)-2-oxo-2,3-dihydro-1H-indol-3-

ylidene]methyl}-1H-pyrrol-3-yl)propanoic

acid

282.29 C16H14N2O3 20 nM

3. SU5416

Semaxinib

NH

O

NH

CH3

H3C

(3Z)-3-[(3,5-dimethyl-1H-pyrrol-2-yl)

methylidene]-2,3-dihydro-1H-indol-2-one

238.28 C15H14N2O 1µM

4. SU6668

NH

O

HO

H3C NH

O

CH3

310.35 C18H18N2O3 1.73

µM



Sr.

No.


formula

IC50*

(3Z)-[(2-oxo-2,3-dihydro-1H-indol-3-

ylidene)methyl-1H-pyrrol-3-yl]propanoic

acid

5. SU11248

Sunitinib

(Sutent)

HN HN

CH3

CH3

HN

O

N

CH3

H3C

O

N-[2-(diethylamino)ethyl]-2,4-dimethyl-5-

{[(3Z)-2-oxo-2,3-dihydro-1H-indol-3-

ylidene]methyl}-1H-pyrrole-3-

carboxamide

380.48 C22H28N4O2 9 nM

6. ZD4190

NN

N

O

NN

OH3C

NH

Br

F

N-(4-bromo-2-fluorophenyl)-6-methoxy-7-

[2-(1H-1,2,3-triazol-1-

yl)ethoxy]quinazolin-4-amine

459.27 C19H16BrF

N6O2

50 nM

7. ZD6474

Vandetanib NH3C

O

O

HN

H3C

F

Br

N-(4-bromo-2-fluorophenyl)-7-methoxy-6-

[(1-methylpiperidin-4-yl)methoxy]

naphthalene-1-amine

473.38 C24H26BrF

N2O2

40 nM

8. ON III CH3

OCH3HO

H3C

OH O 1-(2,4-dihydroxy-6-methoxy-3,5-dimethyl

phenyl)-3-phenylprop-2-en-1-one

298.33 C18H18O4 20

µM

9. TNP-470

O

OCH3

NH

O

O

O

CH3

O

CH3

CH3

Cl

5-methoxy-4-[2-methyl-3-(3-methylbut-2-

en-1-yl)oxiran-2-yl]-1-oxaspiro[2.5]octan-

6-yl N-(2-chloroacetyl)carbamate

401.88 C19H28ClN

O6

220

nM



Sr.

No.


formula

IC50*

10. CB676475 N

N

HN

F

Cl

O

O

H3C

H3C N-(4-chloro-2-fluorophenyl)-6,7-

dimethoxy quinazolin-4-amine

333.74 C16H13ClF

N3O2

1.5

µM

11. YM359445

HN

NH

N

N

NO

S

N

CH3

O (4-methylpiperazin -1-yl)methyl]-1,2-

dihydroquinolin-2-yl}-6-[(1E)-[(1,3-

thiazol-4-ylmethoxy)imino]methyl]-2,3-

dihydro-1H-indol-2-one

514.64 C28H30N6O2

S

8.5

nM

12. AZD2171

Cediranib

N

NO

O

CH3

N

O

HN

CH3

F

4-[(4-fluoro-2-methyl-1H-indol-5-yl)oxy]-

6-methoxy-7-[3-(pyrrolidin-1-

yl)propoxy]quinazoline

450.51 C25H27FN4

O3

5 nM

13. BAY43-

9006

Sorafenib

O

NNH

NH

Cl

F3C

O NH

CH3

O

4-{4-[4-chloro-3-(trifluoromethyl)phenyl]

carbamoylamino}phenoxy-N-methyl

pyridine-2-carboxamide

464.82 C21H16ClF3

N4O3

9 nM

14. AGO

13736

Axitinib

NH

N

N

S

NH

H3C

O

N-methyl-2-{3-[(E)-2-(pyridin-2-yl)ethenyl]-

1H-indazol-6-ylsulfanyl}benzamide

386.47 C22H18N4O

S

0.2

nM



Sr.

No.


formula

IC50*

15. RO

4383596 N

N N

N

NH

OCH3

OH 3-[3-(4-methoxyphenyl)-7-(phenylamino)-

1H,2H,3H,4H-pyrimido[4,5-d][1,3]diazin-

1-yl]cyclopentan-1-ol

417.50 C24H27N5O2 30 nM

16. AMG-706

Motesanib

N NH

O

HN NH

N

H3CCH3

N-(3,3-dimethyl-2,3-dihydro-1H-indol-6-

yl)-2-[(pyridin-4-ylmethyl)amino]pyridine-

3-carboxamide

373.45 C22H23N5O

3 nM

17. CEP-7055

N

NH

O

H3C

O

(H3C)2NO

CH3

3-[6-(dimethylamino)-5-oxohexyl]-7-

[(propan-2-yloxy)methyl]-3,13-

diazahexacyclotricosa-nonaen-14-one

523.67 C33H37N3O3

18 nM

18. E7080 NO

O

H2N

CH3

Cl

NH

HN O

O

4-{3-chloro-4-[(cyclopropylcarbamoyl)

amino]phenoxy}-7-methoxy quinoline-6-

carboxamide

426.85 C21H19ClN4

O4

4 nM



Sr.

No.


formula

IC50*

19. GW-

786034

Pazopanib

N

N

N

NH

CH3

H2NO2S

H3C NN

CH3

CH3

5-({4-[(2,3-dimethyl-2H-indazol-6-yl)-

(methyl)amino]pyrimidin-2-yl}amino)-2-

methylbenzene-1-sulfonamide

437.52 C21H23N7O2

S

30 nM

20. SU14813

NH

H3C

H3C

NH

N

O

NH

F

O

O

O

5-[(5-fluoro-2-oxo-2,3-dihydro-1H-indol-

3-yl)methyl]-2,4-dimethyl-N-[3-

(morpholin-4-yl)-2-oxopropyl]-1H-

pyrrole-3-carboxamide

442.48 C23H27FN4

O4

50 nM

21. BAY 57-

9352

Telatinib

N N

O

NHO

N

O

Cl

NH

H3C

4-[({4-[(4-chlorophenyl)amino]furo[2,3-

d]pyridazin-7-yl}oxy)methyl]-N-methyl

pyridine-2-carboxamide

409.83 C20H16ClN5

O3

19 nM

22. KRN-951

Tivozanib

O

Cl

NH

NH

O

N

N

H3CO

H3CO

O

H3C 1-{2-chloro-4-[(6,7-dimethoxyquinolin-4-

yl)oxy]phenyl}-3-(5-methyl-1,2-oxazol-3-

yl)urea

454.86 C22H19ClN4

O5

0.16

nM

23. ABT-869

Linifanib

NH

N

HN

HNH3C

F

H2N

O

375.40 C21H18FN5

O

4 nM



Sr.

No.


formula

IC50*

3-[4-(3-amino-1H-indazol-4-yl)phenyl]-1-

(2-fluoro-5-methylphenyl)urea

24. OSI-930 HN

OCF3

S

NH

N

O

3-[(quinolin-4-ylmethyl)amino]-N-[4-(tri

fluoromethoxy)phenyl]thiophene-2-

carboxamide

443.44 C22H16F3N3

O2S

9 nM

25. CP-

547,632

Br

F

F

O

NS

NH2

NH

NH

N

O

O

3-[(4-bromo-2,6-difluorophenyl)methoxy]-

5-({[4-(pyrrolidin-1-yl)butyl]carbamoyl}

amino)-1,2-thiazole-4-carboxamide

532.40 C20H24BrF2

N5O3S

11 nM

26. BIBF-1120

Vargatef HN

N

NH

OH3C

H3C

NO

O

O

NH3C

methyl 3-[({4-[N-methyl-2-(4-methyl

piperazin-1-yl)acetamido]phenyl} amino)

phenyl)methyl]-2-oxo-2,3-dihydro-1H-

indole-6-carboxylate

541.64 C31H35N5O4 34 nM

27. BMS-

582664

Brivanib

NN

NO

H3C O

HN

CH3

FH3C

O

NH2

H3C

O

1-({4-[(4-fluoro-2-methyl-1H-indol-5-

yl)oxy]-5-methylpyrrolo[2,1-

f][1,2,4]triazin -6-yl}oxy)propan-2-yl 2-

aminopropanoate

441.46 C22H24FN5

O4

25 nM



Sr.

No.


formula

IC50*

28. CHIR-258

Dovitinib N

NH

N

NCH3

HN

O

F

NH2

4-amino-5-fluoro-3-[6-(4-methylpiperazin-

1-yl)-1H-1,3-benzodiazol-2-yl]-1,2-

dihydroquinolin-2-one

392.43 C21H21FN6

O

10 nM

29. AEE-788

NNH

HN CH3

N

N

H3C

2-{4-[(4-ethylpiperazin-1-

yl)methyl]phenyl }-N-(1-phenylethyl)-1H-

pyrrolo[2,3-b]pyridin-4-amine

439.60 C28H33N5 77 nM

30. CHIR-265

or RAF265

O

N

NH

NHN

N

CF3

HN

F3C

H3C

1-methyl-5-({2-[4-(trifluoromethyl)-1H-

imidazol-2-yl]pyridin-4-yl}oxy)-N-[4-(tri

fluoromethyl)phenyl]-2,3-dihydro-1H-1,3-

benzodiazol-2-amine

520.43 C24H18F6N6

O

30

µM

31. ZK-

304709

(MTGI)

NH

H3CCH3

NH

H2NO2S

O 3-(3,3-dimethyl-2,3-dihydro-1H-indol-2-

ylidene)-2-oxo-2,3-dihydro-1H-indole-5-

sulfonamide

355.41 C18H17N3O3

S

30 nM

32. AP24534

Ponatinib

H3C

N

NN

HN

O CF3

N

N

CH3

532.56 C29H27F3N6

O

1.5

nM



Sr.

No.


formula

IC50*

3-(2-{imidazo[1,2-b]pyridazin-3-

yl}ethynyl)-4-methyl-N-{4-[(4-methyl

piperazin-1-yl)methyl]-3-(trifluoromethyl)

phenyl}benzamide

33. BAY 73-

4506

Regora-

fenib

F3C

N

HNCH3

O

FHN

HN

ClO

1-[4-chloro-3-(trifluoromethyl)phenyl]-3-

[2-fluoro-4-({2-[(methylamino)methyl]

pyridin-4-yl}oxy)phenyl]urea

482.82 C21H15ClF4

N4O3

40 nM

34. BMS

794833

N

O

HN NHF

Cl

H2N

F

O

O

N-{4-[(2-amino-3-chloropyridin-4-yl)oxy]-

3-fluorophenyl}-5-(4-fluorophenyl)-4-oxo-

1,4-dihydropyridine-3-carboxamide

468.84 C23H15ClF2

N4O3

< 3

nM

35. BMS

540215

Brivanib

HN

F

O

N

NN

CH3

O

CH3

H3C

OH

1-({4-[(4-fluoro-2-methyl-1H-indol-5-

yl)oxy]-5-methylpyrrolo[2,1-

f][1,2,4]triazin -6-yl}oxy)propan-2-ol

370.38 C19H19FN4

O3

25 nM

36. KI 8751

OCH3

OCH3N

O

F HN

HN

F

FO

1-(2,4-difluorophenyl)-3-{4-[(6,7-

dimethoxyquinolin-4-yl)oxy]-2-fluoro

phenyl}urea

469.41 C24H18F3N3

O4

0.9

nM



Sr.

No.


formula

IC50*

37. KRN 633

H3CO

H3CO

N

N

O

Cl HN

HN

CH3

O

1-{2-chloro-4-[(6,7-dimethoxyquinazolin-

4-yl)oxy]phenyl}-3-propylurea

416.86 C20H21ClN4

O4

160

nM

38. NVP-

BHG712 CF3

N

NN

N

HN

CH3N

H3C

HN

O

4-methyl-3-{[1-methyl-6-(pyridin-3-yl)-

1H-pyrazolo[3,4-d]pyrimidin-4-yl]amino}-

N-[3-(trifluoromethyl)phenyl]benzamide

503.48 C26H20F3N7

O

4.2

nM

39. PD 173074 OCH3

OCH3N

N NNH

N

CH3

CH3

NH

NH

CH3

H3C CH3

O

3-tert-butyl-1-(2-{[4-(diethylamino)butyl]

amino}-6-(3,5-dimethoxyphenyl)pyrido

[2,3-d]pyrimidin-7-yl)urea

523.67 C28H41N7O3 100

nM

40. XL184

N

HN

NH

N

O

O NH

HN

F

O

O

O

1-N-(4-fluorophenyl)-1-N'-{4-[(2-{[2-

(morpholin-4-yl)ethyl]carbamoyl}-1H-

pyrrolo[2,3-b]pyridin-4-yl)oxy]phenyl}

cyclopropane-1,1-dicarboxamide

586.61 C31H31FN6

O5

0.035

nM



Sr.

No.


formula

IC50*

41. XL880

Foretinib

H3CO

NON

O

O

NH

HN

F

O

O

F

1-N'-[3-fluoro-4-({6-methoxy-7-[3-

(morpholin-4-yl)propoxy]quinolin-4-

yl}oxy)phenyl]-1-N-(4-fluorophenyl)cyclo

propane-1,1-dicarboxamide

632.65 C34H34F2N4

O6

0.8

nM

* IC50 values for VEGFR-2 inhibition kinase assay

Table 4.2: List of some Anti-Angiogenic 1,3-diarylpropenone analogues reported in US Patent

6906105 (J. Phillip Bowen et al 2002)

Sr.

No.

Molecule Structure and IUPAC Name Mol.

Weight

Mol.

formula

%

inhibition*

1. PT-1

O 1,3-Diphenyl-prop-2-en-1-one

208 C15H12O 94.4

2. PT-2 OCH3H3CO

O 1,3-Bis-(4-methoxy-phenyl)propenone

268.31 C17H16O3 85.2

3. PT-3 O

1,5-Diphenyl-penta-1,4-dien-3-one

234.29 C17H14O

96.8

4. PT-8 O

O 2-Benzylidene-indan-1,3-dione

234.25 C16H10O2 34

5. PT-9

CH3

CH3

O

O

3-Benzylidene-pentane-2,4-dione

188.22 C12H12O2

82.7



Sr.

No.


Weight

Mol.

formula

%

inhibition*

6. PT-18

O

CF3

3-phenyl-1-[3-(trifluoromethyl)

phenyl]prop-2-en-1-one

276.25 C16H11F3O 96.9

7. PT-19

O

Cl Cl

1,3-Bis-(4-chloro-phenyl)-propenone

277 C15H10Cl2O

31.1

8. PT-20 CH3H3C

O 1,3-bis(4-methylphenyl)prop-2-en-1-

one

236.31 C17H16O 89.5

9. PT-22 CH3

O CH3 1-(2,4-Dimethyl-phenyl)-3-phenyl-

propenone

236.31 C17H16O 87.2

10. PT-23

OCH3

H3C

3-(2,4-Dimethyl-phenyl)-1-phenyl-

propenone

236.31 C17H16O 89.6

11. PT-25 O

Cl

O

4-benzoyloxy-4'-chlorochalcone

362 C22H15ClO3

62.9

12. PT-26

O

H3C

CH3

CH3

4-isopropyl-4’-methylchalcone

264.36 C19H20O

59.2

13. PT-27 CH3Cl

Cl O 3-(2,6-dichlorophenyl)-1-(4-methyl

phenyl)prop-2-en-1-one

291 C16H12Cl2O 84.6

14. PT-28 CH3

Cl

Cl

O 3-(2,4-Dichloro-phenyl)-1-p-tolyl-

propenone

291 C16H12Cl2O 98.1



Sr.

No.


Weight

Mol.

formula

%

inhibition*

15. PT-29

O

Cl CH3

1-(4-Chloro-phenyl)-3-p-tolyl-

propenone

256 C16H13ClO

58.5

16. PT-30

O

H3C Cl

3-(4-Chloro-phenyl)-1-p-tolyl-

propenone

256 C16H13ClO

46.5

17. PT-31

O

Cl

Cl 3-(2,6-dichlorophenyl)-1-phenylprop-

2-en-1-one

277 C15H10Cl2O 97.5

18. PT-32

O

Cl

Cl

Cl

Cl 1,3-Bis-(2,6-dichloro-phenyl)-

propenone

346.03 C15H8Cl4O 97.0

19. PT-33

O

Cl

Cl 1-(2,6-dichlorophenyl)-3-phenylprop-

2-en-1-one

277.14 C15H12Cl2O 88.7

20. PT-34

O

F

F

F

F

F

F

F

F

F

F

1,3-Bis-pentafluorophenyl-propenone

388.16 C15H2F10O 88.6

21. PT-35

O

F

F

F

F

F

3-Pentafluorophenyl-1-phenyl-

propenone

298 C15H7F5O 0.0

22. PT-36

OF

F

F

F

F

1-Pentafluorophenyl-3-phenyl-

propenone

298.21 C15H7F5O 88.7

23. PT-37

O

OMe

OMe

MeO

OMe 1,3-Bis-(2,6-dimethoxy-phenyl)-

328.36 C19H20O5

39.2



Sr.

No.


Weight

Mol.

formula

%

inhibition*

propenone

24. PT-38

OOCH3

OCH3

1-(2,6-Dimethoxy-phenyl)-3-phenyl-

propenone

268 C17H16O3 63.5

25. PT-39

O

H3CO

OCH3 3-(2,6-Dimethoxy-phenyl)-1-phenyl-

propenone

268 C17H16O3 60.8

26. PT-40

O

Cl

Cl

OMe

OMe 1-(2,6-Dichloro-phenyl)-3-(2,6-

dimethoxy-phenyl)-propenone

336 C17H14Cl2O3 52.3

27. PT-41 OBr

2-Bromo-1,3-diphenyl-propenone

287 C15H11BrO 79.8

28. PT-45

O

O 3-(furan-2-yl)-1-phenylprop-2-en-1-

one

198.22 C13H10O2 0.0

29. PT-46

N N

O 1,3-Di-pyridin-2-yl-propenone

210 C13H10N2O 85.2

30. PT-47

N

O 1-Phenyl-3-pyridin-2-yl-propenone

209 C14H11NO

96.9

31. PT-48

N

O 1-Phenyl-3-pyridin-3-yl-propenone

209 C14H11NO

83.2

32. PT-49 O

HN

3-Phenyl-1-(1H-pyrrol-2-yl)-

propenone

197 C13H11NO

19.4



Sr.

No.


Weight

Mol.

formula

%

inhibition*

33. PT-50 OHN

1-Phenyl-3-(1H-pyrrol-2-yl)-

propenone

197 C13H11NO

0.0

34. PT-51

O

1-(9-anthryl)-3-phenyl-2-propenone

308 C23H16O 69

35. PT-52

O

3-(9-anthryl)-1-phenyl-2-propenone

308 C23H16O 71.3

36. PT-53

O

1,3-di-(9-anthryl)-2-propenone

408 C27H18O 4.7

37. PT-54

O

1-(9-anthryl)-3-(2-naphthyl)-2-

propenone

358 C27H18O

48.1

38. PT-56 O

1-Biphenyl-4-yl-3-phenyl-propenone

284.35 C21H16O 0.0

39. PT-57 O

3-Biphenyl-4-yl-1-phenyl-propenone

284.35 C21H16O 41.4



Sr.

No.


Weight

Mol.

formula

%

inhibition*

40. PT-58 O

1-(1-naphthyl)-3-phenyl-2-propenone

258 C19H14O 68

41. PT-59

Cl O

Cl

3-(2,6-Dichloro-phenyl)-1-naphthalen

-2-ylpropenone

327.20 C19H12Cl2O 97.4

42. PT-60 O

1,3-Di-naphthalen-1-yl-propenone

308 C23H16O 20.3

43. PT-61

O 1-(2-naphthyl)-3-phenyl-2-propenone

258.31 C19H14O 94.7

44. PT-62

O 3-(naphthalen-2-yl)-1-phenylprop-2-

en-1-one

258.31 C19H14O 44.3

45. PT-68

O 2-Benzylidene-3,4-dihydro-2H-

naphthalen-1-one

234.29

C17H14O 97.5

46. PT-70

O 2-Naphthalen-2-ylmethylene-3,4-

dihydro-2H-naphthalen-1-one

284.35 C26H16O

23.6

47. PT-72 OF

Cl

F

Cl 1,5-Bis-(2-chloro-6-fluoro-

phenyl)penta-1,4-dien-3-one

339.16 C17H10Cl2F2O 86.6



Sr.

No.


Weight

Mol.

formula

%

inhibition*

48. PT-73 OCl

Cl

Cl

Cl 1,5-Bis-(2,6-dichloro-phenyl)penta-

1,4-dien-3-one

372.07 C17H10Cl4O 90.4

49. PT-75

O 2,6-Dibenzylidenecyclohexanone

274.14 C20H18O 92.8

50. PT-79

O

N N

2,6-Bis-pyridin-2-ylmethylene-

cyclohexanone

276.33 C18H16N2O 96.7

* Percent inhibition at 6µg/ml density of cells in SVR cell growth inhibition assay

Plot of number of molecules vs. molecular weight and Log P were prepared for all known

angiogenesis inhibitors (Table 4.1) and patented angiogenesis inhibitors (Table 4.2) for the

comparison study.

4.2.4 Study of binding site of VEGFR-2

Characteristics of binding site of VEGFR-2 receptor were studied using Sitemap module.

The co-crystallized ligand present in the PDB structure was identified and deleted. The study

involved following three steps -

1. Finding binding site

2. Mapping of binding site

3. Evaluation of binding site for various parameters

The active site was evaluated in terms of a number of properties like Site Score, Exposure

and Enclosure, Hydrophobic and Hydrophilic character, Contact and Donor/Acceptor

character on the basis of site points and the grids generated in the mapping stage. Five maps

i.e. hydrophilic, hydrophobic, donor, acceptor and surface were generated for further study

of the receptor.

4.2.5 Design of test molecules

• Sets of approximately 400 test molecules were designed (Fig. 4.1) based on chalcone

molecules reported as angiogenesis inhibitors in US patents and articles.



Ar Ar'

O

R

H

Chalcone analogue

Ar, Ar' -Substituted Phenyl, Heterocyclic

R - H, Alkyl

Figure 4.1: Designed 1,3-diarylpropenone analogues

• Various optimizations strategies were used for designing of test molecules – (Figure 4.2)

� Bio-isosteric replacement of various functional groups

� Homologation of alkyl chains

� Aromatic ring substitution by addition of electron withdrawing or electron donating

groups or combinations of both at various positions

� Chain extensions or elongations

Figure 4.2: Various backbone structures of designed molecules

O

R5

R2

R3

R4

R6

A B

R1

R1, R2, R3, R4, R5, R6: -H, -OH, -NH2, -OR, -NHR, -OCOR, -NHCOR, -OCOAr, -NHCOAr, -X, -NO2

R5

R2

R3

R4

R6

A B

R1 O

R2

R3

R4

A B

R1 O

R5 R6

R - Alkyl, Ar - Phenyl, Substituted phenyl, heterocyclic

O

R2

R3

R4

R8

A B

R1

R5

R6

R7

R9

R2

R3

R4

R8

A B

R1

R5

R6

R7

R9

O

R1, R2 , R3, R4, R5, R6 , R7, R8, R9: -H, -OH, -NH2, -OR, -NHR, -OCOR,

-NHCOR, -OCOAr, -NHCOAr, -X, -NO2

R - Alkyl, Ar - Phenyl, Substituted phenyl, heterocyclic Ar1 Ar2

O

Ar1 Ar2

O

Ar1, Ar2: Substituted - Naphthyl, Furyl, Thenyl, Pyridyl, furyl,

Indolyl, Quinolyl, Phenyl, Pyrolyl, Pyrazolyl, Benzofuryl, Cyclohexyl

Ar

O

R2

R1

R3

R4

ArR2

OR1

R3

R4

R1, R2, R3, R4: Alkyl, -OH, -OR, -OCOR, -NH2, -NHR

Ar: Substituted - Phenyl, Naphthyl, heterocyclic rings



4.2.6 Ligand Preparation

The three dimensional structures of 42 known VEGFR-2 inhibitors (Molecules in markets

and clinical trials), 50 patented molecules reported to be angiogenesis inhibitors, 990 decoy

molecules and 400 test molecules were prepared using the LigPrep module of Maestro in the

Schrödinger suite of tools.

The bond orders of these ligands were fixed and the ligands ‘cleaned’ through LigPrep

specifying a pH value of 7.0. There is a possibility that tautomeric forms interacts differently

with the binding site and one of the tautomer interacts more strongly relative to the other

forms. Hence, most probable tautomers of the compounds were chosen based on their

interactions with the proteins in the X-ray structures. In the final stage of LigPrep, the

compounds were energy minimized with Merck Molecular Force Field (MMFF)

4.2.7 Validation of docking protocol

Before carrying out docking studies with the database of designed ligands, the performance

of the docking protocol was evaluated with the poses obtained for co-crystallized ligands in

cognate (self) docking study or re-docking study.

SP Precision Glide docking procedure was validated by removing native ligand from the

binding site and re-docking it in the binding site of receptor.

For example, ligand of PDB structure 3BE2 (Fig. 4.3) was extracted, energy minimized with

LigPrep and docked in the active site of 3BE2 receptor structure.

O

NH

HN

N

N

N

CH3

CF3

NH3C

CH3

Figure 4.3: Structure of co-crystallized ligands of PDB structures - 3BE2 (http://www.pdb.org

/pdb/explore/explore.do?structureId=3BE2)

The validation process consisted of evaluation of two parameters:

(i) Retention of key interactions seen between ligand and receptor in the native X-ray

complex

(ii) RMSD values of less than 2 A° between the top-ranked docked pose and the X-ray

pose of co-crystallized ligand.



4.2.8 Enrichment study

In order to identify VEGFR-2 protein structure suitable for computational study, enrichment

of active inhibitors using docking study were carried out on the ‘database’ of 1000 ligands

(10 actives seeded amongst 990 decoy molecules from binding database,

http://www.bindingdb.org/bind/ByTargetNames.jsp) in the active site of the protein. This

database has a random hit rate of 1%.

Docking study on database of actives and decoys was carried out in the active site of

VEGFR-2 (PDB – 3BE2, 1Y6B, 2P2H) structures using the Schrödinger docking program,

Glide.

The active inhibitors belonged to seven distinct chemotypes and spanned a molecular weight

range of 400-500 daltons. The decoy set of molecules also spanned the similar range of

molecular weight and had at least 150 different chemotypes represented in it. The property

distribution indicated that there was enough diversity in the decoys as well as actives

considered in the study.

The top ranked pose for each docked ligand (based on Glide-score) was saved. Percentages

of actives retrieved in the top 5% (50) and 10% (100) of the database ranks were counted.

For each hit, only the model/pose with the lowest Glide Score was retained. This was

followed by the counting of actives retrieved from the top 50 (5%) and 100 (10%) ranks.

The study helped to identify the optimal protocol for the highest enrichment of actives in the

top 5 and 10% of the database by Glide Score ranks

The actives used in the study were SU6668, SU11248, AGO13736, E7080, KRN951,

AZD2171, BAY43-9006, OSI930, CHIR258 and BMS794833. The molecules had seven

different backbone structures (Figure 4.4).

4.2.9 Docking using Glide

The molecules were docked into the binding site of VEGFR-2 (PDB code: 3BE2,

resolution=1.75 A°). The best docking conditions which succeeded to retrieve the pose of the

co-crystallized ligand were used.

1) The known VEGFR-2 inhibitors were docked into the structure of receptor for study of

binding characteristics of those molecules.

2) The patent reported chalcone analogues as angiogenesis inhibitors were docked into

VEGFR-2 structure for studying their interactions within the active site.



Figure 4.4: Representative chemical backbone structures of ligands (A-G) used as actives

(VEGFR-2 inhibitors) in enrichment studies

3) The test molecules were then docked into the structure of VEGFR-2 to check the

interactions with the receptor and for prioritization of molecules having optimum binding

with receptor for synthesis.

In the current docking experiment, the following docking parameters were employed

4.2.9.1. Receptor grid generation

After ensuring that the protein and ligands were in the correct form for docking, a receptor-

grid was generated using the grid-receptor generation program. To soften the potential for

non-polar parts of the receptor, van der Waal radii of receptor atoms were scaled by 0.80 A°

with a partial atomic charge of 0.15.

The protein–ligand complex prepared as described above was employed to build energy

grids within a cubic box of dimensions 15 A° × 15 A°× 15 A° centered around the centroid

of the X-ray ligand pose. Site points were generated followed by generation of the grid

NH

R

H3C

NH

O

CH3

NH

N

N

SR

NH3CO

R1

O Cl

NH

HN

R

O

N

NH3CO

OR

O

HN

O

NNH

NH

O R

O

HN

N

N

NH

HN

O

N

O

HN NH

O

O

(A) (B) (C)

(D) (E)

(F) (G)



displaying the active site with an enclosing box at the centroid of the workspace. The best

docked pose (with lowest Glide Score value) of each molecule was saved per ligand.

4.2.9.2 Docking

Step I: The basic options for docking of ligands consisted of specifying the receptor grid,

selection of the precision method (HTVS, SP or XP), setting flexibility options, setting for

the selection of initial poses and for the energy minimization of the poses

Flexible ligand docking method was opted to generate conformations of ligands internally

during the docking process and default settings related to poses of ligands and energy

minimization stage of docking algorithm were retained.

Step II: Ligands to be used for docking process were specified and all other default options

related to van der Waal radii scaling were retained. No constraints were specified in initial

docking experiments.

4.2.9.2.1 Primary filter

In order to determine the criteria score for primary filter, ligands (Figure 4.5) whose binding

modes are already known (PDB ID: 3BE2, Y6B, 2P2H, 2QU6, 3CP9, 3CPC and 3CPB)

were docked into structure of VEGFR-2.

About 400 designed ligands were then docked into the binding site of VEGFR-2 using

GLIDE-SP method and ranked by G-Score. The compounds with G-Score lower than 8.0

were removed from the set of 400 designed molecules.

Concerning the evaluation of isomers, all the isomers were selected as ‘hits’, in case at least

one of the isomers had score higher than 8.0.

In this process, about 230 out of 400 compounds were defined as primary ‘hits’ (57.50 %).

4.2.9.2.2 Secondary filter

In the binding mode prediction, Standard Precision mode of Glide was used to predict the

binding mode of the compounds identified as primary ‘hits’. Initially, several ligands the

binding modes of which were known (Figure 4.5) were used to find out the binding

requirements for ligands with VEGFR-2. The primary hits which showed similar binding

characteristics were retained in the set.



Additional inhibitors from the literature, the binding modes of which are not known (known

angiogenesis inhibitors, set of 41 compounds, Table 4.1) were docked into the binding site of

VEGFR-2. Then the binding free energies were calculated in order to determine the criteria

of binding free energy for the hit selection.

Figure 4.5: PDB Ligand structures used in docking study

For 115 compounds, the calculated binding energy was lower than the criteria determined.

These compounds were classified as secondary ‘hits’, which comprised approximately 50 %

of primary hits

O

NH

HN

N

N

N

CH3

CF3

NH3C

CH3

3BE2

H3CO

HN

SO2N

O

N

HN

1Y6B

N

N

N

HNH3CO

H3CO

OCH3

N

HN

2P2H

N

HNN

Cl

O

O

O

NH

CH3

ON

2QU6

N

NH2N

N

H3C

O

HN CH3

CH3

3CP9

NH

O

H3C

O

NH

H2N

N

3CPB

N

H2N

N

O

H3C N

F

FF

3CPC

CH3



4.2.9.3 Visualization of docking results

The glide score (G-score), glide energy value, H-bonds and van der Waals contacts (good,

bad and ugly) to the receptor were visualized in the Glide pose viewer using default settings

to analyze the binding modes of the ligands to receptor.

4.2.10 Screening of designed molecules for in silico prediction of ADME properties

The QikProp program was used to obtain the ADME properties of the analogues. All the

analogues were neutralized before being used by QikProp. The program was processed in

normal mode, and predicted 44 properties for the 115 test molecules (secondary hits),

consisting of physically significant descriptors and physiochemical properties with a detailed

analysis of the log P (Octanol/Water), and log HERG.

It also evaluated the acceptability of the analogues based on Lipinski’s rule of 5, which is

essential for rational drug design.

The prediction of high probability of success or failure using the rule is based on drug

likeness for molecules complying with 2 or more of the rules namely- molecular mass less

than 500 Dalton, high lipophilicity expressed as Log P less than 5, less than 5 hydrogen bond

donors and less than 10 hydrogen bond acceptors.

4.2.11 Screening of designed molecules for in silico prediction of toxicity potential

Toxic properties like mutagenicity, carcinogenicity, skin irritation and eye irritation were

predicted in silico using Toxtree software for 115 designed molecules (secondary hits).

4.2.12 Prioritization of designed molecules for synthesis

4.2.12.1 Tertiary filter

ADMET properties were predicted for secondary hits obtained in docking study. The

molecules with suitable physiochemical and pharmacokinetic properties with no possible

toxicity hazards were prioritized for synthesis.

Hence, 50 molecules were selected as tertiary hits (43.47 %) out of 115 secondary hits. The

fifteen molecules were then prioritized from the tertiary hits for synthesis and evaluation for

anti-angiogenic activity. Chalcone and hydroxylated naphthyl chalcone (D9) were selected

for synthesis for comparison study in addition to fifteen compounds. (Table 4.3)



Table 4.3: List of designed molecules prioritized for synthesis (Bowen et al, 2002)

Sr.

No.

Compound

code

Structure and IUPAC Name MW Mol

formula

1 Chalcone

O 1,3-Diphenylprop-2-en-1-one

208 C15H12O

2 D9

O

OH

1-(3-Hydroxy-phenyl)-3-(naphthalen-2-yl)prop-

2-en-1-one

274 C19H14O2

3 D10

O

NH

O

N-[3-(3-Naphthalen-2-yl- prop-2-

enoyl)phenyl]benzamide

377 C26H19NO2

4 DM1

O

NH

O

OCH3

OCH3

OCH3

O2N 4-Nitro-N-{3-[3-(3,4,5-trimethoxyphenyl)prop-

2-enoyl]phenyl}benzamide

462 C25H22N2O7

5 DM3

O

OCH3

OCH3

OCH3

O

O2N

O

3-[3-(3,4,5- trimethoxyphenyl)prop-2-

enoyl]phenyl 4-nitrobenzoate

463 C25H21NO8

6 DM5

O

NH

O2N

OOCH3

N-{3-[3-(4-Methoxy-naphthalen-1-yl)prop-2-

enoyl]-phenyl}-4-nitro-benzamide

452 C27H20N2O5

7 DM7

O

NH

OCH3

OCH3

OCH3

O

N-{3-[3-(3,4,5- trimethoxyphenyl)prop-2-

enoyl]phenyl}naphthalene-2-carboxamide

467 C29H25NO5

8 DM8

O

NH

N(CH3)2O

N-{3-[3-(4-(dimethylamino)phenylprop-2-

enoyl]phenyl}naphthalene-2-carboxamide

420 C28H24N2O2

9 E3

O

O

C

O

3-[3-(naphthalen-2-yl-)prop-2-

enoyl]phenylnaphthalene 2-carboxylate

428 C30H20O3



Sr.

No.

Compound

code

Structure and IUPAC Name MW Mol

formula

10 E5

O

O

C

O

CF3 3-[3-(naphthalen-2-yl-)prop-2-enoyl]phenyl 3-

(trifluoromethyl)benzoate

446 C27H17F3O3

11 F3

NH

C

O

O

N-{3-[3-(naphthalene-2-yl)prop-2-enoyl]-

phenyl}naphthalene-2-carboxamide

427

C30H21NO2

12 F5

NH

C

O

O

CF3 N-{3-[3-(naphthalen-2-yl-)prop-2-

enoyl]phenyl}-3-(trifluoromethyl)benzamide

445 C27H18

F3NO2

13 F7

NH

H2C

ONO2

3-(naphthalene-2-yl)-1-(3-{[(4-nitrophenyl)

methyl]amino}phenyl)prop-2-en-1-one

408 C26H20N2O3

14 G10

NH

NO2

O

O

N-{3-[3-(naphthalen-2-yl)-3-oxoprop-1-en-1-

yl]phenyl}-4-nitrobenzamide

422 C26H18N2O4

15 J20

NH

H3COO

O

N-{3-[3-(4-methoxynaphthalen-1-yl)-3-oxoprop-

1-en-1-yl]phenyl}benzamide

407 C27H21NO3

16 PG1

O

NH

OH

OCH3

OCH3

O2N

O

N-{3-[3-(3-Hydroxy-4,5-dimethoxy-

phenyl)prop-2-enoyl]phenyl}-4-nitro-benzamide

448 C24H20N2O7

17 PG4

O

NH

O2N

OOH

N-{3-[3-(4-Hydroxy-naphthalen-1-yl)prop-2-

enoyl]-phenyl}-4-nitro-benzamide

438

C26H18N2O5



B. SYNTHESIS OF DESIGNED 1,3-DIARYLPROPENONE ANALOGUES

This section consists of –

I. General procedures for preparation of 1,3-diarylpropenone analogues

II. Synthesis of designed analogues of 1,3-diarylpropenone analogues

I. General procedures for preparation of 1,3-diarylpropenone analogues

4.3 Methods and Instruments

All reactions were monitored by thin layer chromatography (TLC) using thin layer

aluminium plate Merck pre-coated silica gel GF254 of 0.2 mm thickness for completion of

reaction and for establishing purity of synthesized compounds. The spots were viewed either

under short ultraviolet (UV) light (254 nm) and/or long UV light (365 nm) using Expo Hi

Tech UV cabinet.

Column chromatography was performed using Qualigens 60-120 mesh silica gel for

purification of synthesized compounds.

The melting point of the individual compound was recorded with Veego VMP III melting

point apparatus. The reported melting points of compounds were obtained from literature

(Vogel et al, 1989, Susan, 2006)

Other instruments used for the study were Shimadzu BL 220H balance, rotary evaporator

and vacuum oven.

The raw materials, intermediates and the final compounds were characterized by IR

Spectroscopy, Mass spectrometry and 1H NMR spectroscopy

Infrared (IR) spectra were recorded on a Perkin Elmer FT-IR spectrophotometer as KBr

pellets.

1H NMR was recorded using Bruker Advance spectrometer (300 MHz) with DMSO d6 and

CDCl3 as the solvent. Chemical shifts were reported in ppm down field from

tetramethylsilane as the internal standard.

The Infrared and 1H-NMR spectra were interpreted using standard textbooks.

The molecular weights were determined by gas chromatography-mass spectrometry (GC-

MS).

4.4 Chemicals and Solvents

All solvents and reagents used for the study were of general reagent grade.

The starting materials used in this study were commercially available benzaldehyde (1),

acetophenone (2), 2-naphthaldehyde (4), 3-hydroxyacetophenone (5), 3-amino acetophenone



(7) and 3,4,5-trimethoxybenzaldehyde (13), 4-hydroxyacetophenone (15) and N,N-

dimethylaminobenzaldehyde (25).

Other chemicals used were benzoyl chloride (8), 4-nitrobenzoyl chloride (11), 2-naphthoyl

chloride (22), 3-trifluoromethylbenzoyl chloride (29), 4-nitrobenzyl chloride (35), sodium

hydroxide (NaOH), phosphorus oxychloride (POCl3) and anhydrous magnesium sulphate.

The organic solvents used were ethanol, methanol, ethyl acetate, hexane, dichloromethane

(DCM), acetone, chloroform, N,N-dimethylformamide (DMF) and dimethyl sulphoxide

(DMSO).

4.5 General scheme of synthesis of 1,3-diarylpropenone analogues

The general scheme for synthesis of 1,3-diarylpropenone analogues is as follows (Scheme

4.1)

Scheme 4.1: Synthesis of 1,3-diarylpropenone analogues

4.6 Preparation of 1,3-diarylpropenone analogues

Substituted aldehydes were reacted with substituted ketones with alpha hydrogen atom in

presence of either base or acid catalyst in an alcoholic solvent at room temperature (RT) or

reflux temperature to give the desired 1,3-diarylpropenone analogues.

Appropriately substituted ketones were prepared by aroylation and alkylation reactions.

Substituted aldehydes were prepared by formylation reaction.

4.6.1 Preparation of substituted Ketones

4.6.1.1 Synthesis of substituted ketone using aroylation

Substituted acetophenone was dissolved in sufficient DCM and stirred well. Pyridine was

added to it and the reaction mixture was cooled in an ice bath. Substituted aroyl chloride was

added drop wise to the reaction mixture and the temperature of the reaction mixture was

allowed to come to room temperature. The reaction mixture was then stirred at RT for

Ar H

O

Ar' CH2

O

R

Ar Ar'

O

R

H

Base / Acid

Substituted

Aldehyde

Substituted Ketone

with alpha hydrogen

1,3-Diarylpropenone

analogues

Ar, Ar' - Substituted Phenyl, Heterocyclic

R - H, Alkyl



suitable time period. The reaction was monitored by TLC. After the completion of reaction,

solvent was evaporated and the slurry was poured in ice water. The precipitated product was

filtered and washed with water until neutral to litmus and then dried.

4.6.1.2 Synthesis of substituted ketone using alkylation

To the solution of substituted hydroxy acetophenone in acetone, sodium carbonate was

added followed by addition of sodium iodide. Substituted benzyl chloride was then added

and the reaction mixture was stirred at room temperature. The reaction was monitored by

TLC. After the completion of reaction, the solvent was evaporated and the mixture was

poured into ice water. The precipitated product was filtered and washed with water until

neutral to litmus and then dried.

4.6.2 Preparation of substituted Aldehydes

4.6.2.1 Synthesis of substituted aldehyde using Formylation

To a solution of substituted naphthalene in DMF, POCl3 was added under stirring. The

mixture was heated to 80°C and reaction was monitored using TLC. After the completion of

reaction, the reaction mixture was poured in ice water. The precipitated product was washed

with water until neutral to litmus and dried.

4.6.3 Preparation of 1,3-diarylpropenone analogues

Three methods were used for preparation of 1,3-diarylpropenone analogues using base or

thionyl chloride. Initially, fifteen 1,3-diarylpropenone analogues were synthesized, out of

which two analogues were further subjected to demethylation to give total of seventeen 1,3-

diarylpropenone analogues.

4.6.3.1 Synthesis of substituted 1,3-diarylpropenone analogues using base

A solution of substituted acetophenone in ethanol was added to substituted benzaldehyde in

a 100 ml conical flask. The mixture was stirred with a magnetic stirrer and 40% NaOH was

added drop wise into it. The mixture was stirred at room temperature and then kept overnight

in refrigerator. The reaction was monitored by TLC periodically. After the completion of

reaction, the mixture was poured into ice water and allowed to stand until complete

precipitation of the product. The product was filtered and washed with water until neutral to

litmus. It was dried and purified by recrystallization or column chromatography (Solvent

systems: Hexane: Ethyl acetate or Chloroform).



4.6.3.2 Synthesis of substituted 1,3-diarylpropenone analogues using thionyl chloride

A solution of substituted acetophenone in methanol was added to solution of substituted

benzaldehyde in methanol in a 100 ml round bottom flask. The mixture was stirred with a

magnetic stirrer and thionyl chloride was added to it. The reaction mixture was refluxed for

suitable time and monitored by TLC. After the completion of reaction, the mixture was

poured into ice water and allowed to stand until complete precipitation of the product. The

product was filtered and washed with water until neutral to litmus. It was dried and purified

by column chromatography technique (Solvent systems: Hexane: Ethyl acetate or

Chloroform).

4.6.3.3 Synthesis of substituted 1,3-diarylpropenone analogues using demethylation

The synthesized 1,3-diarylpropiophenone was dissolved in DCM in 100 ml conical flask.

Aluminium chloride was added and the reaction mixture was stirred at RT. The reaction was

monitored by TLC. After the completion of reaction, the reaction mixture was poured into

mixture of dilute hydrochloric acid and ice. The organic layer was separated and washed

with water to neutral pH. The organic layer was then dried over anhydrous magnesium

sulfate and then distilled to get the product. The product was then purified by column

chromatography (Solvent systems: Hexane: Ethyl acetate or Chloroform).

II. Synthesis of designed analogues of 1,3-diarylpropenone

4.7 Synthesis of Chalcone (3)

4.7.1 Step I: Preparation of 1,3-diphenylprop-2-en-1-one (3) from benzaldehyde (1) and

acetophenone (2)

+

COCH3CHO

O

NaOH

Ethanol

1 2 3

Scheme 4.2: Synthesis of chalcone (3)

Using procedure 4.6.3.1, reaction of 1 (1.76 g, 0.0166 mol) with 2 (2 g, 0.0166 mol) in 24

hrs yielded the product 3.

Yield: 2.5 g (72 %)



4.8 Synthesis of D9 (6)

4.8.1 Step I: Preparation of 1-(3-Hydroxyphenyl)-3-(naphthalen-2-yl)prop-2-en-1-one (6)

from 2-naphthaldehyde (4) and 3-hydroxyacetophenone (5)

CHO

+

O

OH

COCH3

OH

NaOH

Ethanol

4 5 6

Scheme 4.3: Synthesis of D9 (6)

Using procedure 4.6.3.1, reaction of 4 (1 g, 0.0064 mol) with 5 (0.87 g, 0.0064 mol) in 24


Yield: 0.8 g (45.71 %)

4.9 Synthesis of D10 (10)

4.9.1 Step I: Preparation of 3-benzoylaminoacetophenone (9) from 3-aminoacetophenone

(7)

Scheme 4.4: Synthesis of 3-benzoylaminoacetophenone (9)

Using procedure 4.6.1.1, benzoylation of 7 (5 g, 0.037 mol) using benzoyl chloride (8) (6.19

g, 0.044 mol) and pyridine (3.48 g, 0.044 mol) in 1 hr gave the product 9.

Yield: 8 g (90.39 %)

4.9.2 Step II: Preparation of N-[3-(3-Naphthalen-2-yl-prop-2-enoyl)phenyl]benzamide (10)

from 2-naphthaldehyde (4) and 3-benzoylaminoacetophenone (9)

O

NH

O

COCH3

NH

OCHO

+NaOH

Ethanol

4 9 10

Scheme 4.5: Synthesis of D10 (10)



Yield: 1.5 g (61.98 %)

COCH3

NH

O

COCH3

NH2

+

COCl

Pyridine

DCM, RT

7 8 9



4.10 Synthesis of DM1 (14)

4.10.1 Step I: Preparation of 3-(4-nitrobenzoyl)aminoacetophenone (12) from 3-amino

acetophenone (7)

Scheme 4.6: Synthesis of 3-(4-nitrobenzoyl)aminoacetophenone (12)

Using procedure 4.6.1.1, benzoylation of 7 (2.5 g, 0.0185 mol) with 4-nitrobenzoyl chloride

(11) (4.08 g, 0.022 mol) and pyridine (1.74 g, 0.022 mol) in 1 hr gave the product 12.

Yield: 4.5 g (85.5 %)

4.10.2 Step II: Preparation of 4-Nitro-N-{3-[3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]

phenyl}benzamide (14) from 3-(4-nitrobenzoyl)aminoacetophenone (12) and 3,4,5-

trimethoxybenzaldehyde (13)

Scheme 4.7: Synthesis of DM1 (14)

Using procedure 4.6.3.1, reaction of 12 (0.89 g, 0.0031 mol) with 13 (0.5 g, 0.0031 mol) in

24 hrs yielded the product 14.

Yield: 0.9 g (64.28 %)


4.11.1 Step I: Preparation of 4-(4-nitrobenzoyl)oxyacetophenone (16) from 4-hydroxy

acetophenone (15)

Scheme 4.8: Synthesis of 4-(4-nitrobenzoyl)oxyacetophenone (16)

COCH3

NH

NO2

O

COCH3

NH2

+

COCl

NO2

Pyridine

DCM, RT

7 11 12

CHO

OCH3

OCH3

H3CO

COCH3

NH

NO2

O

+

O

NH

O

OCH3

OCH3

OCH3

O2N

1312 14

Ethanol

NaOH

OH3COC

O

NO2

COCH3

OH

COCl

NO2

+Pyridine

DCM, RT

1511 16



Using procedure 4.6.1.1, benzoylation of 15 (5 g, 0.0367 mol) with 4-nitrobenzoyl chloride

(11) (8.16 g, 0.044 mol) and pyridine (3.48 g, 0.044 mol) in 1 hr at RT gave the product 16.

Yield: 10.3 g (98.47 %)

4.11.2 Step II: Preparation of 3-[3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]phenyl-4-nitro

benzoate (17) from 3,4,5-trimethoxybenzaldehyde (13) and 4-(4-nitrobenzoyl)oxy

acetophenone (16)

O

OCH3

OCH3

OCH3

O

O2N

O

CHO

OCH3

OCH3

H3CO

+OH3COC

O

NO2

SOCl2

Ethanol, Reflux

13 16 17




Yield: 0.8 g (49.38 %)


4.12.1 Step I: Preparation of 1-methoxynaphthalene (19) from 1-naphthol (18)

OH

Dimethyl sulphate

OCH3

18 19

Na2CO3

Scheme 4.10: Synthesis of 1-methoxynaphthalene

To a solution of 18 (5 g, 0.0347 mol) in acetone, sodium carbonate (7.35 g, 0.0694 mol) was

added under stirring followed by addition of dimethyl sulphate (8.75 g, 0.0694 mol). The

mixture was refluxed and reaction was monitored using TLC. After the completion of

reaction, solvent was evaporated and the slurry was poured in ice water. The precipitated

product 19 was washed with water until neutral to litmus and dried. The product was then

purified by column chromatography (Hexane: Ethyl acetate)

Yield: 3.84 g (70 %)

4.12.2 Step II: Preparation of 4-methoxy-1-naphthaldehyde (20) from 1-

methoxynaphthalene (19)



CHO

OCH3

OCH3

Formaldehyde

POCl3, DMF

19 20

Scheme 4.11: Synthesis of 4-methoxy-1-naphthaldehyde (20)

Using procedure 4.6.2.1, Formylation of 19 (3 g, 0.0189 mol) with POCl3 (5.78 g, 0.0378

mol) and DMF (3.45 g, 0.04725 mol) in 3 hrs gave the product 20. The synthesized product

was then purified by column chromatography (Hexane: Ethyl acetate)

Yield: 2.4 g (67.98 %)

4.12.3 Step III: Preparation of 3-(4-nitrobenzoyl)aminoacetophenone (12) from 3-amino

acetophenone (7) – As given in Scheme 4.6, Page No. 55

4.12.4 Step IV: Preparation of N-{3-[3-(4-Methoxynaphthalen-1-yl)prop-2-enoyl]phenyl}-

4-nitrobenzamide (21) from 4-methoxy-1-naphthaldehyde (20) and 3-(4-nitrobenzoyl)

aminoacetophenone (12)

O

NH

O2N

OOCH3

COCH3

NH

CHO

OCH3

O

NO2

2012 21

Ethanol

NaOH+


Using procedure 4.6.3.1, reaction of 12 (0.6 g, 0.0021 mol) with 20 (0.39 g, 0.0021 mol) in


Yield: 0.6 g (63.15 %)


4.13.1 Step I: Preparation of 3-naphthoylaminoacetophenone (23) from 3-

aminoacetophenone (7) and 2-naphthoyl chloride (22)

COCH3

NH

O

COCH3

NH2

+

COCl Pyridine

DCM, RT

7 22 23

Scheme 4.13: Synthesis of 3-naphthoylaminoacetophenone (23)



Using procedure 4.6.1.1, reaction of 7 (2 g, 0.0148 mol) using 2-naphthoyl chloride (22)

(3.37 g, 0.0177 mol) and pyridine (1.4 g, 0.0177 mol) in 2 hrs at RT gave the product 23.

Yield: 3.9 g (97.5 %)

4.13.2 Step II: Preparation of N-{3-[3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]phenyl}

naphthalene-2-carboxamide (24) from 3-naphthoylaminoacetophenone (23) and 3,4,5-

trimethoxybenzaldehyde (13)

O

NH

OCH3

OCH3

OCH3

O

COCH3

NH

O

CHO

H3CO

OCH3

OCH3

+NaOH

Ethanol

13 23 24


Using procedure 4.6.3.1, reaction of 13 (1 g, 0.0051 mol) with 23 (0.816 g, 0.0051 mol) in


Yield: 1 g (42 %)


4.14.1 Step I: Preparation of 3-naphthoylaminoacetophenone (23) from 3-

aminoacetophenone (7) – As given in Scheme 4.13, Page No. 57

4.14.2 Step II: Preparation of N-{3-[3-(4-(dimethylamino)phenylprop-2-enoyl]phenyl}

naphthalene-2-carboxamide (26) from 3-naphthoylaminoacetophenone (23) and 4-(N,N-

dimethyl)aminobenzaldehyde (25)

O

NH

N(CH3)2O

COCH3

NH

O(H3C)2N

CHO+

NaOH

Ethanol

2523 26




Yield: 1.5 g (53.57 %)



4.15 Synthesis of E3 (28)

4.15.1 Step I: Preparation of 3-(2-naphthoyl)oxyacetophenone (27) from 3-hydroxy

acetophenone (5) and 2-naphthoyl chloride (22)

Scheme 4.16: Synthesis of 3-(2-naphthoyl)oxyacetophenone (27)

Using procedure 4.6.1.1, aroylation of 5 (2 g, 0.0147 mol) using 2-naphthoyl chloride (22)

(3.35 g, 0.0176 mol) and pyridine (1.39 g, 0.0176 mol) in 2 hrs at RT gave the product 27.

Yield: 4.2 g (98.36 %)

4.15.2 Step II: Preparation of 3-[3-(naphthalen-2-yl)prop-2-enoyl]phenylnaphthalene-2-

carboxylate (28) from 2-naphthaldehyde (4) and 3-(2-naphthoyl)oxyacetophenone (27)

Scheme 4.17: Synthesis of E3 (28)



Yield: 1.8 g (65.7 %)

4.16 Synthesis of E5 (31)

4.16.1 Step I: Preparation of 3-(3-trifluoromethyl)oxyacetophenone (30) from 3-hydroxy

acetophenone (5)

Scheme 4.18: Synthesis of 3-(3-trifluoromethyl)oxyacetophenone (30)

COCH3

O

O

COCH3

OH

+

COCl Pyridine

DCM, RT

22 275

COCH3

O

O

COCH3

OH

+

COCl

CF3

Pyridine

DCM, RT CF3

5 29 30

O

O

C

O

COCH3

O

OCHO

+SOCl2

Ethanol, Reflux

27 284



Using procedure 4.6.1.1, aroylation of 5 (1 g, 0.0074 mol) using 3-trifluoromethyl benzoyl

chloride (29) (1.83 g, 0.0088 mol) and pyridine (0.7 g, 0.0088 mol) in 2 hrs gave the product

30.

Yield: 2.2 g (97.34 %)

4.16.2 Step II: Preparation of 3-[3-(naphthalen-2-yl)prop-2-enoyl]phenyl-3-(trifluoro

methyl)benzoate (31) from 2-naphthaldhyde (4) and 3-(3-trifluoromethyl)oxyacetophenone

(30)

O

O

C

O

CF3

COCH3

O

O

CF3

CHO

+SOCl2

Ethanol, Reflux

4 30 31

Scheme 4.19: Synthesis of E5 (31)

Using procedure 4.6.3.2, reaction of 4 (0.5 g, 0.0032 mol) with 29 (0.99 g, 0.0032 mol) in 2


Yield: 1 g (69.93 %)

4.17 Synthesis of F3 (32)

4.17.1 Step I: Preparation of 3-naphthoylaminoacetophenone (23) from 3-aminoaceto

phenone (7) – As given in Scheme 4.13 Page No. 57

4.17.2 Step II: Preparation of N-{3-[3-(naphthalene-2-yl)prop-2-enoyl]phenyl}

naphthalene-2-carboxamide (32) from 2-naphthaldehyde (4) and 3-naphthoylaminoaceto

phenone (23)

NH

C

O

O

COCH3

NH

OCHO

+NaOH

Ethanol

4 23 32

Scheme 4.20: Synthesis of F3 (32)



Yield: 2.5 g (91.57 %)




4.18.1 Step I: Preparation of 3-(3-trifluoromethyl)aminoacetophenone (33) from 3-amino

acetophenone (7)

Scheme 4.21: Synthesis of 3-(3-trifluoromethyl)aminoacetophenone (33)

Using procedure 4.6.1.1, aroylation of 7 (1 g, 0.0074 mol) using 3-trifluoromethylbenzoyl

chloride (29) (1.83 g, 0.0088 mol) and pyridine (0.7 g, 0.0088 mol) in 2 hrs gave the product

33.

Yield: 2 g (88.1 %)

4.18.2 Step II: Preparation of N-{3-[3-(naphthalen-2-yl-)prop-2-enoyl]phenyl}-3-

(trifluoromethyl)benzamide (34) from 2-naphthaldehyde (4) and 3-(3-

trifluoromethyl)aminoacetophenone (33)

NH

C

O

O

CF3

COCH3

NH

O+

CF3

CHOSOCl2

Ethanol, Reflux

4 33 34




Yield: 2 g (70.17 %)


4.19.1 Step I: Preparation of 3-(4-nitrobenzyl)aminoacetophenone (36) from 3-amino

acetophenone (7)

Scheme 4.23: Synthesis of 3-(4-nitrobenzyl)aminoacetophenone (36)

COCH3

NH

O

COCH3

NH2

+

COCl

CF3

Pyridine

DCM, RT CF3

7 29 33

COCH3

NH

NO2

COCH3

NH2

+

CH2Cl

NO2

Na2CO3, NaI

Acetone, RT

7 35 36



Using procedure 4.6.1.2, alkylation of 7 (2 g, 0.0148 mol) using 4-nitrobenzyl chloride (35)

(3.03 g, 0.0177 mol), sodium iodide (2.21 g, 0.0148 mol) and sodium carbonate (6.27 g,

0.0592 mol) in 3 hrs gave the product 36.

Yield: 3 g (75 %)

4.19.2 Step II: Preparation of 3-(naphthalen-2-yl)-1-(3-{[(4-nitrophenyl)methyl]

amino}phenyl)prop-2-en-1-one (37) from 2-naphthaldehyde (4) and 3-(4-nitrobenzyl)

aminoacetophenone (36)

NH

H2

C

ONO2

COCH3

NH

NO2

CHO

+

NaOH

Ethanol

4 36 37


Using procedure 4.6.3.1, reaction of 4 (0.5 g, 0.0032 mol) with 36 (0.87 g, 0.0032 mol) in 24


Yield: 0.8 g (61.5 %)

4.20 Synthesis of G10 (38)

4.20.1 Step I: Preparation of 3-(4-nitrobenzoyl)aminoacetophenone (12) from 3-amino

acetophenone (7) – As given in Scheme 4.6 Page No. 55

4.20.2 Step II: Preparation of N-{3-[3-(naphthalen-2-yl)-3-oxoprop-1-en-1-yl]phenyl}-4-

nitrobenzamide (38) from 2-naphthaldehyde (4) and 3-(4-nitrobenzoyl)amino acetophenone

(12)

NH

NO2

O

O

COCH3

NH

NO2

O+

CHO NaOH

Ethanol

4 12 38

Scheme 4.25: Synthesis of G10 (38)



Yield: 1 g (67.11 %)



4.21 Synthesis of J20 (39)

4.21.1 Step I: Preparation of 1-methoxynaphthalene (19) from 1-naphthol (18) – As given

in

Scheme 4.10, Page No. 56

4.21.2 Step II: Preparation of 4-methoxy-1-naphthaldehyde (20) from 1-

methoxynaphthalene (19) – As given in Scheme 4.11, Page No. 57

4.21.3 Step III: Preparation of 3-benzoylaminoacetophenone (9) from 3-

aminoacetophenone (7) – As given in

Scheme 4.44.4, Page No. 54

4.21.4 Step IV: Preparation of N-{3-[3-(4-methoxynaphthalen-1-yl)-3-oxo-prop-1-en-1-

yl]phenyl}benzamide (39) from 3-benzoylaminoacetophenone (7) and 4-methoxy-1-

naphthaldehyde (20)

NH

H3COO

CHO

OCH3

+

COCH3

NH

O

O

NaOH, Ethanol

207 39

Scheme 4.26: Synthesis of J20 (39)



Yield: 1.3 g (59.63 %)

4.22 Synthesis of PG1 (40)

4.22.1 Step I: Preparation of N-{3-[3-(3-Hydroxy-4,5-dimethoxyphenyl)prop-2-enoyl]

phenyl}-4-nitrobenzamide (40) from 4-Nitro-N-{3-[3-(3,4,5-trimethoxyphenyl)prop-2-

enoyl]phenyl}benzamide (14)

O

NH

O

OCH3

OCH3

OCH3

O2N O

NH

O

OCH3

OCH3

OH

O2N

AlCl3

DCM, RT

14 40

Scheme 4.27: Synthesis of PG1 (40)

Using procedure 4.6.3.3, reaction of 14 (0.5 g, 0.0010 mol) with aluminium chloride (2 g,

0.0015 mol) in 8 hrs yielded the product 40.



Yield: 0.15 g (31.25 %)

4.23 Synthesis of PG4 (41)

4.23.1 Step I: Preparation of N-{3-[3-(4-hydroxy-naphthalen-1-yl)prop-2-enoyl]phenyl}-4-

nitrobenzamide (41) from N-{3-[3-(4-methoxynaphthalen-1-yl)prop-2-enoyl]phenyl}-4-

nitrobenzamide (21)

O

NH

O2N

OOCH3

O

NH

O2N

OOH

AlCl3

DCM, RT

21 41

Scheme 4.28: Synthesis of PG4 (41)

Using procedure 4.6.3.3, reaction of 21 (0.5 g, 0.0011 mol) with aluminium chloride (0.22 g,

0.0017 mol) in 10 hrs yielded the product 41.

Yield: 0.2 g (41.66 %)



C. PROFILES OF REACTANTS AND SYNTHESIZED COMPOUNDS

The section consists of –

I. Profiles of Reactants used in the study

II. Profiles of Intermediates synthesized in the study

III. Profiles of Products prepared in the study

I. Profiles of Reactants used in the study

4.24 Compound: 1

IUPAC Name: Benzaldehyde

Molecular Formula: C7H6O

Molecular Weight: 106.04

Physical Data:

• State: Liquid

• Color: Clear and colorless

• Boiling Point: 178 °C (Lit. 176-178°C) (Budavari, 2006)

Table 4.4: Interpretation of IR spectrum (KBr disk) of benzaldehyde (1) [Fig. 8.1, Page 176]

Sr. No. Wave number cm-1

Peaks Group Assignment

1. 3068.70 C–H Stretching Aromatic

2. 2878.68 C–H Stretching Aldehyde


4. 1691.69 C=O Stretching Aldehyde

5. 1617.67, 1468.27 C–C Stretching Aromatic

6. 728.54, 690.21 C–H Bending Mono substituted benzene

4.25 Compound: 2

IUPAC Name: Acetophenone



Physical Data:

• State: Liquid

• Color: Clear and colorless

• Boiling Point: 202 °C (Lit. 202 °C) (Budavari, 2006)

Table 4.5: Interpretation of IR spectrum (KBr disk) of acetophenone (2) [Fig 8.2, Page 176]




2. 2878.68 C–H Stretching Methyl Ketone

3. 1663.69 C=O Stretching Ketone

OHC

H3COC






5. 1377.36 C–H Bending Methyl Ketone


4.26 Compound: 4

IUPAC Name: 2-Naphthaldehyde



Physical Data:

• State: Solid

• Color: White

• Melting Point: 58-60 °C (Lit. 57-60 °C) (Budavari, 2006)

Table 4.6: Interpretation of IR spectrum (KBr disk) of 2-naphthaldehyde (4) [Fig 8.3, Page

177]




2. 2828.81, 2711.48 C–H Stretching Aldehyde




4.27 Compound: 5

IUPAC Name: 3-Hydroxyacetophenone

Molecular Formula: C8H8O2


Physical Data:

• State: Solid

• Color: White

• Melting Point: 94-96 °C (90-95 °C) (Budavari, 2006)

Table 4.7: Interpretation of IR spectrum (KBr disk) of 3-hydroxyacetophenone (5) [Fig 8.4,

Page 177]



1. 3173.87 O–H Stretching Phenol





6. 1424.71 C-O-H Bending Phenol


CHO

OH

COCH3





8. 1261.38 C-O Stretching /

O-H bending

Phenol

9. 910.96, 793.79, 681.82 C–H Bending 1,3-disubstituted benzene

4.28 Compound: 7

IUPAC Name: 3-Aminoacetophenone

Molecular Formula: C8H9ON


Physical Data:

• State: Solid

• Color: White


Table 4.8: Interpretation of IR spectrum (KBr disk) of 3-aminoacetophenone (7) [Fig 8.5, Page

178]



1. 3466.02, 3363.01 N–H Stretching Primary amine






7. 1321.70 C-N Stretching Aromatic amine


4.29 Compound: 13

IUPAC Name: 3,4,5-trimethoxybenzaldehyde



Physical Data:

• State: Solid

• Color: White

• Melting Point: 76 °C (73-75 °C) (Budavari, 2006)

Table 4.9: Interpretation of IR spectrum (KBr disk) of 3,4,5-trimethoxybenzaldehyde (13) [Fig

8.6, Page 178]




2. 2942.34 C–H Stretching Methyl ether

3. 2970.75 C–H Stretching Alkyl



NH2

COCH3

CHO

OCH3

OCH3H3CO






7. 1391.76 C-H Bending Alkyl

8. 1233.51, 1127.64 C-O Stretching Aromatic ether

9. 845.34 C–H Bending 1,3,4,5-substituted benzene

4.30 Compound: 15

IUPAC Name: 4-Hydroxyacetophenone



Physical Data:

• State: Solid

• Color: White

• Melting Point: 100 °C (Lit. 100-104 °C) (Budavari, 2006)

Table 4.10: Interpretation of IR spectrum (KBr disk) of 4-Hydroxyacetophenone (15) [Fig 8.7,

Page 179]










8. 1216.17 C-O Stretching Phenol

9. 865.68, 793.55 C–H Bending 1,4-disubstituted benzene

4.31 Compound: 18

IUPAC Name: 1-hydroxynaphthalene



Physical Data:

• State: Solid

• Color: Pink

• Melting Point: 96 °C (Lit. 95-96 °C) (Budavari, 2006)

Table 4.11: Interpretation of IR spectrum (KBr disk) of 1-hydroxynaphthalene (18) [Fig 8.8,

Page 179]





COCH3

OH

OH









4.32 Compound: 25

IUPAC Name: 4-(N,N-dimethylamino)benzaldehyde

Molecular Formula: C9H11NO


Physical Data:

• State: Solid

• Color: White


Table 4.12: Interpretation of IR spectrum (KBr disk) of 4-(N,N-dimethylamino) benzaldehyde

(25) [Fig 8.9, Page 180]








6. 1596.01 C–C Stretching Aromatic

7. 1369.91 C-N Stretching Aromatic amine


CHO

N(CH3)2



II. Profiles of Intermediates synthesized in the study

4.33 Compound: 9

IUPAC Name: 3-Benzoylaminoacetophenone

Molecular Formula: C15H13NO2


Physical Data:

• State: Solid

• Color: White

• Melting Point: 114 °C

Table 4.13: Interpretation of IR spectrum (KBr disk) of 3-Benzoylaminoacetophenone (9) [Fig

8.10, Page 180]



1. 3323.16 N–H Stretching Secondary amide



4. 1667.32 C=O Stretching Ketone, Secondary Amide


6. 1547.53 N-H Bending Secondary amide



9. 865.40, 791.04, 698.88 C-H Bending 1,3-disubstituted benzene

4.34 Compound: 12

IUPAC Name: 3-(4-nitrobenzoyl)aminoacetophenone

Molecular Formula: C15H12N2O4


Physical Data:

• State: Solid

• Color: Light Yellow


Table 4.14: Interpretation of IR spectrum (KBr disk) of 3-(4-nitrobenzoyl)amino acetophenone

(12) [Fig 8.11, Page 181]






4. 1668.60 C=O Stretching Ketone, secondary amide


6. 1548.67, 1344.77 N=O Stretching Aromatic nitro group


NH

COCH3

NO2

O

NH

COCH3

O








Table 4.15: Mass Fragmentation of 3-(4-nitrobenzoyl)aminoacetophenone (12) [Fig 8.37 and

8.38, Page 194]

Sr.

No.

m/z value of characteristics product ions Interpretations

Positive ionization

1. 307.2 M+23 i.e. M+Na

2. 285.2 M+H

3. 286.1 M+2H

Negative ionization

4. 283.2 M-H

4.35 Compound: 16

IUPAC Name: 4-(4-nitrobenzoyl)oxyacetophenone



Physical Data:

• State: Solid

• Color: Light brown


Table 4.16: Interpretation of IR spectrum (KBr disk) of 4-(4-nitrobenzoyl)oxy acetophenone

(16) [Fig 8.12, Page 181]





3. 1746.00 C=O Stretching Ester




7. 1179.32 C-O Stretching Ester


4.36 Compound: 19

IUPAC Name: 1-Methoxynaphthalene



Physical Data:

OCH3

H3COC

O

O

NO2



• State: Solid

• Color: Pale Yellow

• Boiling Point: 138 °C (Lit. 135-137 °C)

Table 4.17: Interpretation of IR spectrum (KBr disk) of 1-Methoxynaphthalene (19) [Fig 8.13,

Page 182]









4.37 Compound: 20

IUPAC Name: 4-Methoxy-1-naphthaldehyde



Physical Data:

• State: Solid

• Color: White

• Melting Point: 36°C (Lit. 35-36°C)

Table 4.18: Interpretation of IR spectrum (KBr disk) of 4-Methoxy-1-naphthaldehyde (20) [Fig

8.14, Page 182]






4. 1681.66 C=O Stretching Aromatic Aldehyde


6. 1396.24 C–H Bending Aldehyde


8. 815.25 C–H Bending 1,4-Substituted aromatic

Table 4.19: Interpretation of 1H NMR spectrum (DMSO) of 4-Methoxy-1-naphthaldehyde (20)

[Fig 8.73, Page 212]

Sr. No. Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 10.18 s 1H - CO

Ar

H

2. 9.23 d 1H 8.1 Aromatic proton at C-8

CHO

OCH3

1

2

345

6

7

8



Sr. No. Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

3. 8.29-8.26 d 1H 8.4 Aromatic proton at C-5


5. 7.72-7.76 t 1H 8.1 Aromatic proton at C-7



8. 4.10 s 3H - Ar-OCH3

4.38 Compound: 23

IUPAC Name: 3-(2-naphthoyl)aminoacetophenone



Physical Data:

• State: Solid

• Color: Off white


Table 4.20: Interpretation of IR spectrum (KBr disk) of 3-(2-naphthoyl)amino acetophenone

(23) [Fig 8.15, Page 183]













4.39 Compound: 27

IUPAC Name: 3-(2-naphthoyl)oxyacetophenone



Physical Data:

• State: Solid


NH

COCH3

O

O

COCH3

O




Table 4.21: Interpretation of IR spectrum (KBr disk) of 3-(2-naphthoyl)oxyacetophenone (27)

[Fig 8.16, Page 183]












4.40 Compound: 30

IUPAC Name: 3-(3-triflouromethylbenzoyl)oxyacetophenone

Molecular Formula: C16H11F3O3


Physical Data:

• State: Solid

• Color: Light brown


Table 4.22: Interpretation of IR spectrum (KBr disk) of 3-(3-triflouromethylbenzoyl)oxy

acetophenone (30) [Fig 8.17, Page 184]










8. 1122.39 C-F Stretching Fluoro compound


4.41 Compound: 33

IUPAC Name: 3-(3-triflouromethylbenzoyl)aminoacetophenone

Molecular Formula: C16H12F3NO2


Physical Data:

• State: Solid

O

COCH3

CF3

O

NH

COCH3

O

CF3





Table 4.23: Interpretation of IR spectrum (KBr disk) of 3-(3-triflouromethylbenzoyl)amino

acetophenone (33) [Fig 8.18, Page 184]








6. 1542.97 N-H bending Secondary amide


8. 1125.99 C-F Stretching Fluoro compound


4.42 Compound: 36

IUPAC Name: 3-(4-nitrobenzyl)aminoacetophenone

Molecular Formula: C8H9ON


Physical Data:

• State: Solid

• Color: Yellow


Table 4.24: Interpretation of IR spectrum (KBr disk) of 3-(4-nitrobenzyl)amino acetophenone

(30) [Fig 8.19, Page 185]



1. 3342.03 N–H Stretching Secondary amine



4. 1667.59 C=O Stretching Aromatic Ketone


6. 1583.12 N-H Bending Secondary amine

7. 1518.88 N=O Stretching Aromatic nitro group


9. 1340.13 C-N Stretching

N=O Stretching

Aromatic amine

Aromatic nitro group

10. 861.83 C–H Bending 1,4-disubstituted benzene


NH

COCH3

NO2



III. Profiles of Products prepared in the study

4.43 Compound: Chalcone (3)

IUPAC Name: 1,3-diphenyl-prop-2-en-1-one


Molecular Weight: 208

Physical Data:

• State: Solid

• Color: White

• Melting Point: 56-58 °C

Table 4.25: Interpretation of IR spectrum (KBr disk) of 1,3-diphenyl-prop-2-en-1-one (3) [Fig

8.20, Page 185]






4. 1578.67 C=C Stretching Alkene


6. 968.06 C–H Bending Trans Alkene

7. 760.07, 699.69 C–C Bending Aromatic

Table 4.26: Mass Fragmentation of 1,3-diphenyl-prop-2-en-1-one (3) [Fig 8.39 and 8.40, Page

195]

Sr.

No.


Positive ionization

1. 231.4 M+Na

2. 209.4 M+H

3. 131.3 M-77

4. 103.3 M-105

5. 105.2 M-103

Negative ionization

6. 207 M-H

Table 4.27: Interpretation of 1H NMR spectrum (DMSO) of 1,3-diphenyl-prop-2-en-1-one (3)

[Fig 8.74, Page 213]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 8.03-8.07 d 1H 12 Ar Ar

O H

2. 7.80-7.95 m 3H - Aromatic protons at C-2’,

O

1

2

3

1 '

2 '3 '

4 '

5 '

6 '

1 "

2 ''

3 ''

4 ''

5 ''

6 ''



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

C-6’ and

Ar Ar

O

H

3. 7.52-7.63 m 4H - Aromatic protons at C-3’, C-

4’, C-5’, C-6”

4. 7.44-7.45 m 1H - Aromatic proton at C-2”

5. 7.36-7.42 t 2H 8.1 Aromatic protons at C-3”, C-

5”

6. 7.07-7.08 m 1H - Aromatic proton at C-4”

4.44 Compound: D9 (6)

IUPAC Name: 1-(3-hydroxy-phenyl)-3-(naphthalen-2-yl)prop-2-en-1-one



Physical Data:

• State: Solid

• Color: Yellow


Table 4.28: Interpretation of IR spectrum (KBr disk) of D9 (6) [Fig 8.21, Page 186]



1. 3385.90 O-H Stretching Phenol

2. 3058.82 C-H Stretching Alkene

3. 1666.03 C=O Stretching α,β-unsaturated Aromatic Ketone

4. 1577.57 C-C Stretching Aromatic



7. 1272.72 C-O Stretching

(O-H Bending)

Phenol

8. 981.81 C-H Bending Trans Alkene

9. 852.37, 819.77, 724.47 C-H Bending 1,3-substituted benzene

10. 749.83 C-H Bending Mono substituted Aromatic Ring

Table 4.29: Mass Fragmentation of D9 (6) [Fig 8.41 and 8.42, Page 196]

Sr.

No.


Positive ionization

1. 297.5 M+Na

2. 275.2 M+H

3. 257.4 M-19

4. 181.3 M-93

5. 153.1 M-121

O

OH1

2

31'

2'

3'

4'5'

6'

1"

2"

3"4"5"

6"

7"8"



Sr.

No.


6. 146.0 M-128

7. 121.2 M-153

Negative ionization

8. 273.0 M-H

Table 4.30: Interpretation of 1H NMR spectrum (DMSO) of D9 (6) [Fig 8.75, Page 214]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 9.85 Broad, s (D2O

exchangeable) 1H - -OH

2. 8.10-8.12 d 1H 8.1 Aromatic protons at C-1”


C-3”, C-4”, C-5”, C-8”

4. 7.85-7.90 d 1H 15.3 Ar Ar

O H

5. 7.50-7.72 m 4H - Aromatic protons at C-6”,

C-7”, C-2’, C-5’

6. 7.38-7.43 d 1H 15.3 Ar Ar

O

H

7. 7.08 d 1H 8.1 Aromatic protons at C-4’

4.45 Compound: D10 (10)

IUPAC Name: N-[3-(3-Naphthalen-2-yl-prop-2-enoyl)phenyl]benzamide



Physical Data:

• State: Solid



Table 4.31: Interpretation of IR spectrum (KBr disk) of D10 (10) [Fig 8.22, Page 186]



1. 3248.27 N-H Stretching Secondary Amide

2. 3055.95 C-H Stretching Aromatic


4. 1644.86 C=O Stretching Secondary Amide

5. 1590.07, 1570.25 C-C Stretching Aromatic



1

3

2

45

6

1'

3'

4'5'

6'

7'

8' 1"

2"

3"

4"

5"

6"2'

NH

O

O






9. 716.94 C-H Bending Mono substituted Aromatic Ring

Table 4.32: Mass Fragmentation of D10 (10) [Fig 8.43 and 8.44, Page 197]

Sr.

No.


Positive ionization

1. 400.3 M+Na

2. 378.3 M+H

3. 257.3 M-120

4. 250.2 M-127

5. 105.2 M-272

6. 77.2 M-300

Negative ionization

7. 376.2 M-H

Table 4.33: Interpretation of 1H NMR spectrum (CDCl3) of D10 (10) [Fig 8.76, Page 215]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 8.47 s 1H - Amide proton -CONH

2. 8.24 s 1H - Aromatic proton at C-2

3. 8.15-8.18 d 1H 8.1 Aromatic proton at C-1’

4. 7.8-8.0 m 9H -

Aromatic protons at C-4, C-6, C-

3’, C-4’, C-5’, C-8’, C-2”, C-6”

and

Ar Ar

O H

5. 7.58-7.63 d 1H 15.6 Ar Ar

O

H

6. 7.4-7.6 m 6H - Aromatic protons at C-3”, C-4”,

C-5”, C-5, C-6’, C-7’

4.46 Compound: DM1 (14)

IUPAC Name:4-Nitro-N-{3-[3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]phenyl}benzamide



Physical Data:

• State: Solid

• Color: Yellow

• Melting Point: >250 °C

O

NH

O

OCH3

OCH3

OCH3

O2N

1

2

3

56

1'2'

6'

1"

2"3"

5"

6" 3'

4'5'

4

4"



Table 4.34: Interpretation of IR spectrum (KBr disk) of DM1 (14) [Fig 8.23, Page 187]



1. 3363.85 N-H Stretching Secondary Amide


3. 2944.19 C-H Stretching Alkyl (-CH3)

4. 2835.09 C-H Stretching Methyl ether

5. 1684.56 C=O Stretching α,β-unsaturated Ketone

6. 1646.18 C=O Stretching Secondary Amide


8. 1570.44 N-H Bending Secondary Amide





13. 975.84 C-H Bending Trans alkene


15. 712.00 C-H Bending 1,4-substituted benzene

Table 4.35: Mass Fragmentation of DM1 (14) [Fig 8.45 and 8.46, Page 198]

Sr.

No.


Positive ionization

1. 485.3 M+Na

2. 463.3 M+H

3. 295.2 M-167

4. 241.2 M-221

5. 193.1 M-269

Negative ionization

6. 461.5 M-H

Table 4.36: Interpretation of 1H NMR spectrum (DMSO) of DM1 (14) [Fig 8.77, Page 216]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton


2. 8.38-8.41 m 3H 9 Aromatic protons at C-3”,

C-5”, C-2

3. 8.21-8.24 d 2H 8.7 Aromatic protons at C-2”,

C-6”

4. 8.11-8.14 d 1H 9 Aromatic proton at C-4


6. 7.83-7.88 d 1H 15.6 Ar Ar

O H



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

7. 7.69-7.75 d 1H 15.3 Ar Ar

O

H


9. 7.24 s 2H - Aromatic protons at C-2’,

C-6’

10. 3.86 s 9H - -OCH3


IUPAC Name: 3-[(2E)-3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]phenyl-4-nitrobenzoate



Physical Data:

• State: Solid

• Color: Yellow






2. 2962.53 C-H Stretching Alkene

3. 2918.76 C-H Stretching Alkyl (CH3)









12. 870.26, 797.68 C-H Bending 1,4-substituted benzene


Sr. No. m/z value of characteristics product ions Interpretations

Positive ionization

1. 486.2 M+ Na

2. 464.3 M+H

3. 270.2 M-193

4. 194.3 M-271

5. 151.1 M-314

O

OCH3

OCH3

OCH3

O

O2N

O1

2

3

45

6

1'

2'

6'

2"

3"

5"

6"

3'4'

5'

1"

4"




6. 150.0 M-313

Negative ionization

7. 462.0 M-H

Table 4.39: Interpretation of 1H NMR spectrum (CDCl3) of DM3 (17) [Fig 8.78, Page 217]

Sr. No. Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 8.4 m 4H - Aromatic protons at C-2”,

C-3”, C-5”, C-6”


3. 7.87-7.88 m 1H - Aromatic proton at C-2

4. 7.73-7.78 d 1H 15.9 Ar Ar

O H



7. 7.35-7.40 d 1H 15.9 Ar Ar

O

H

8. 6.87 s 2H - Aromatic protons at C-2’,

C-6’

9. 3.91 s 9H - -OCH3


IUPAC Name: N-{3-[3-(4-Methoxy-naphthalen-1-yl)prop-2-enoyl]phenyl}-4-nitrobenz

amide



Physical Data:

• State: Solid

• Color: Yellow





1. 3411.02 N-H Stretching Secondary amide




5. 1658.74 C=O Stretching Amide

O

NH

O2N

OOCH3

1

2

3

4

5

6

1'

2'

3'

5'

6'

7'

8'

1"

2"

3"

5"

6"

4"

4'














Sr.

No.


Positive ionization

1. 475.3 M+ Na

2. 453.3 M+H

3. 422.1 M-30

4. 242.4 M-212

5. 183.0 M-269

6. 166.2 M-288

7. 158.4 M-296

8. 122.2 M-330

Negative ionization

9. 451.3 M-H

Table 4.42: Interpretation of 1H NMR spectrum (DMSO) of DM5 (21) [Fig 8.79, Page 218]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton




4. 8.37-8.40 d 2H 8.7 Aromatic proton at C-3”, C-5”

5. 8.22-8.31 m 5H - Aromatic proton at C-2”, C-6”,

C-4, C-6, C-5



8. 7.84-7.89 d 1H 15.3 Ar Ar

O H

9. 7.67-7.72 t 1H 8.1 Aromatic proton at C-6’

10. 7.58-7.63 m 2H - Aromatic proton at C-7’ and



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

Ar Ar

O

H

11. 7.13 d 1H 8.1 Aromatic proton at C-3’

12. 4.06 s 3H - -OCH3


IUPAC Name: N-{3-[3-(3,4,5-trimethoxyphenyl)prop-2-enoyl]phenyl}naphthalene-2-

carboxamide



Physical Data:

• State: Solid

• Color: Yellow

• Melting Point: 190-192 °C









6. 1650.34 C=O Stretching Secondary amide






12. 677.24 C-H Bending Mono substituted benzene


Sr.

No.


Positive ionization

1. 490.3 M+ Na

2. 468.4 M+H

3. 340.4 M-127

4. 312.3 M-155

5. 281 M-186

6. 274.1 M-193

O

NH

OCH3

OCH3

OCH3

O

12

3

45

6

1'

2'

3'

4'5'

6'

1"

2"

3"

4"5"

6"

7"

8"



Sr.

No.


7. 221.3 M-246

8. 155.2 M-312

9. 127.1 M-340

Negative ionization

10. 466.3 M-H


Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton


2. 8.43 s 1H - Aromatic proton at C-1”

3. 7.99-8.22 m 2H - Aromatic protons at C-2, C-3”

4. 7.86-7.98 m 4H - Aromatic protons at C-4”, C-5”,

C-8”, C-4


6. 7.65-7.70 d 1H 15.9 Ar Ar

O H


C-5

8. 7.36-7.41 d 1H 15.9 Ar Ar

O

H

9. 6.78 s 2H - Aromatic proton at C-2’, C-6’

10. 3.88 s 9H - -OCH3


IUPAC Name: N-{3-[3-(4-(dimethylamino)phenylprop-2-enoyl]-phenyl}naphthalene-2-

carboxamide



Physical Data:

• State: Solid

• Color: Yellowish Orange






O

NH

N(CH3)2O

12

3

4

5

6

1'

2'

3'

4'

5'6'

1"

2"

3"

4"5"

6"

7"

8"







4. 1650.34 C=O Stretching α,β-unsaturated Aromatic Ketone,

Amide



7. 1320.96 C-N Stretching Tertiary amine





Sr.

No.


Positive ionization

1. 443.0 M+ Na

2. 421.1 M+H

3. 405.2 M-15

4. 272.1 M-148

5. 246.3 M-174

6. 170.4 M-250

7. 147.2 M-275

8. 121.2 M-301

Negative ionization

9. 419.5 M-H


Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton



3. 8.24 m 2H - Aromatic protons at C-2, C-4”

4. 7.98-8.15 m 6H -

Aromatic protons at C-3”, C-5”,

C-6, C-8”, C-4 and

Ar Ar

O H


C-2’, C-6’, C-5

6. 7.42-7.47 d 1H 15.3 Ar Ar

O

H

7. 6.98-7.12 m 2H - Aromatic proton at C-3’, C-5’

8. 3.2 s 6H - -N(CH3)2



4.51 Compound: E3 (28)

IUPAC Name: 3-[3-(naphthalen-2-yl)prop-2-enoyl]phenylnaphthalene 2-carboxylate



Physical Data:

• State: Solid



Table 4.49: Interpretation of IR spectrum (KBr disk) of E3 (28) [Fig 8.28, Page 189]



1. 3050.34 C-H Stretching Aromatic, Alkene



4. 1600.28 C=C stretching Alkene





9. 775.46, 710.08 C-H Bending Mono substituted benzene

Table 4.50: Mass Fragmentation of E3 (28) [Fig 8.55 and 8.56, Page 203]

Sr.

No.


Positive ionization

1. 451.4 M+ Na

2. 429.3 M+H

3. 301.3 M-127

4. 257.4 M-171

5. 171.2 M-257

6. 155.2 M-273

Negative ionization

7. 427.0 M-H

Table 4.51: Interpretation of 1H NMR spectrum (DMSO) of E3 (28) [Fig 8.82, Page 221]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton



3. 8.07-8.25 m 8H -

Aromatic protons at C-2, C-6,

C-1’, C-5’, C-8’, C-4”, C-5”,

C-8”

O

O

C

O

1

23

4

5

6

1"

2"

3"4"5"

6"

7"

8"

1'

2'

3'

4' 5'

6'

7'

8'



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

4. 7.98-8.14 m 3H - Aromatic protons at C-4, C-5,

C-4’

5. 7.85-7.90 s 1H 15.3 Ar Ar

O H

6. 7.65-7.76 m 4H -

Aromatic protons at C-3’, C-

6’, C-7’ and

Ar Ar

O

H

7. 7.56-7.58 t 2H - Aromatic protons at C-6”,

C-7”

4.52 Compound: E5 (31)

IUPAC Name: 3-[3-(naphthalen-2-yl)prop-2-enoyl]phenyl-3-(trifluoromethyl)benzoate

Molecular Formula: C27H17F3O3


Physical Data:

• State: Solid



Table 4.52: Interpretation of IR spectrum (KBr disk) of E5 (31) [Fig 8.29, Page 190]










8. 1071.94 C-F Stretching -CF3




Table 4.53: Mass Fragmentation of E5 (31) [Fig 8.57 and 8.58, Page 204]


Positive ionization

1. 469.5 M+ Na

2. 447.2 M+H

3. 301 M-145

4. 274.4 M-174

O

O

C

O

CF3

1

23

4

5

6

1'

2'

3'

4'5'

6'

7'

8' 1"

2"

3"

4"

5"

6"




5. 174.3 M-274

Negative ionization

6. 445.4 M-H

Table 4.54: Interpretation of 1H NMR spectrum (CDCl3) of E5 (31) [Fig 8.83, Page 222]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton


2. 8.43 d 1H 7.8 Aromatic proton at C-6”

3. 7.85-8.05 m 8H -


4’, C-5’, C-8’, C-2, C-5, C-6,

C-4”

4. 7.79-7.83 d 1H 13.2 Ar Ar

O H

5. 7.47-7.72 m 6H -


6’, C-7’, C-4, C-5”,

Ar Ar

O

H

4.53 Compound: F3 (32)

Structure

NH

C

O

O

1

23

46

5

1'

2'

3'

4'5'

6'

1"

2"

3"

4" 5"

6"

7"

8"7'

8'

IUPAC Name: N-{3-[3-(naphthalene-2-yl)prop-2-enoyl]phenyl}naphthalene-2-carboxamide



Physical Data:

• State: Solid

• Color: Off White

• Melting Point: >250 °C

Table 4.55: Interpretation of IR spectrum (KBr disk) of F3 (32) [Fig 8.30, Page 190]





3. 1658.74 C=O Stretching α,β-unsaturated Aromatic

Ketone






5. 1586.08 N-H Bending Aromatic Secondary amide







12. 703.31 C-H Bending (Broad) Mono substituted benzene

Table 4.56: Mass Fragmentation of F3 (32) [Fig 8.59 and 8.60, Page 205]

Sr.

No.


Positive ionization

1. 450.3 M+ Na

2. 429.3 M+2H

3. 428.3 M+H

4. 301.4 M-128

5. 274.3 M-153

6. 155.2 M-272

7. 129.2 M-302

Negative ionization

8. 426.3 M-H

Table 4.57: Interpretation of 1H NMR spectrum (DMSO) of F3 (32) [Fig 8.84, Page 223]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton




4. 8.36 s 1H - Aromatic proton at C-1’

5. 8.19-8.23 d 1H 9.6 Aromatic proton at C-3”

6. 7.95-8.14 m 10H -

Aromatic proton at C-4, C-5,

C-6, C-3’, C-4’, C-5’, C-8’, C-

4”, C-5”, C-8”

7. 7.87-7.92 d 1H 15.9 Ar Ar

O H

8. 7.57-7.69 m 5H -

Aromatic proton at C-6”, C-7”,

C-6’, C-7’ and

Ar Ar

O

H




IUPAC Name: N-{3-[3-(naphthalen-2-yl)prop-2-enoyl]phenyl}-3-(trifluoromethyl)

benzamide

Molecular Formula: C27H18 F3NO2


Physical Data:

• State: Solid

• Color: Off White



Sr.

No.

Wave number cm-1










9. 1054.31 C-F Stretching Fluorine compound





Sr.

No.


Positive ionization

1. 468.3 M+ Na

2. 446.3 M+H

3. 318.2 M-127

Negative ionization

4. 444.3 M-H


Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton



3. 8.36 s 2H - Aromatic protons at C-2”, C-1’

NH

C

O

O

CF3

1

23

4

5

6

1'

2'

3'

4'5'

6'

1"

8'

7'

2"

3"

4"

5"

6"



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

4. 8.30-8.33 d 1H 8.1 Aromatic proton at C-6”

5. 8.10-8.2 m 2H - Aromatic protons at C-4, C-4”

6. 7.9-8.06 m 7H -

Aromatic protons at C-5, C-6, C-

3’, C-4’, C-5’, C-8’ and

Ar Ar

O H

7. 7.78-7.83 t 1H 7.5 Aromatic proton at C-5”

8. 7.62-7.66 s 1H 12 Ar Ar

O

H

9. 7.57-7.61 m 2H - Aromatic protons at C-6’, C-7’


IUPAC Name: 3-(naphthalene-2-yl)-1-(3-{[(4-nitrophenyl)methyl]amino}phenyl)prop-2-en-

1-one



Physical Data:

• State: Solid

• Color: Yellow





1. 3342.41 N-H Stretching Secondary amine


3. 2999.42 C-H Stretching Alkyl -CH2-




7. 1518.54 N=O Stretching Aromatic nitro compound


9. 1342.76 C-N Stretching

N=O Stretching

Aromatic secondary amine

Aromatic nitro compound



NH

H2C

ONO2

1

23

4

5

6

1'

2'

3'4'5'

6'

7'

8' 1"

2"3"

4"

5"

6"




Sr.

No.


Positive ionization

1. 431 M+ Na

2. 409 M+H

3. 363.0 M-47

4. 281 M-127

5. 255.2 M-153

6. 137.2 M-271

Negative ionization

7. 407 M-H


Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 8.37-8.40 m 2H 8.2 Aromatic proton at C-3”, C-5”

2. 7.9-8.10 m 2H - Aromatic proton at C-1’, C-8’

3. 7.86-7.91 d 1H 15.2 Ar Ar

O H

4. 7.55-7.60 m 10H -

Aromatic proton at C-3’, C6’,

C-7’, C-2”, C-6”, C-4’, C-5’,

C-6, C-2, and

Ar Ar

O

H

5. 7.20-7.26 m 2H - Aromatic proton at C-4, C-5

6. 4.25 s 2H - -NH-CH2-

7. 3.8 s 1H - -NH-CH2-

4.56 Compound: G10 (38)

IUPAC Name: N-{3-[3-(naphthalen-2-yl)-3-oxoprop-1-en-1-yl]phenyl}-4-nitrobenzamide



Physical Data:

• State: Solid

• Color: Yellow


NH

NO2

O

O

1

2

3

45

6

1'

2'

3'4'5'

6'

7'

8' 1"

2"3"

4"

5"

6"



Table 4.64: Interpretation of IR spectrum (KBr disk) of G10 (38) [Fig 8.33, Page 192]








6. 1584.44, 1347.19 N=O Stretching Aromatic nitro compound

7. 1546.15 C-C Stretching Alkene






Table 4.65: Mass Fragmentation of G10 (38) [Fig 8.65 and 8.66, Page 208]

Sr.

No.


Positive ionization

1. 445.3 M+ Na

2. 423 M+H

3. 393.2 M-31

4. 301.2 M-123

5. 241 M-181

6. 150.1 M-272

7. 121 M-303

8. 77.2 M-345

Negative ionization

9. 421.2 M-H

Table 4.66: Interpretation of 1H NMR spectrum (DMSO) of G10 (38) [Fig 8.87, Page 226]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 10.88 s 1H - - CONH Amide proton


3. 8.37-8.4 d 2H 9.0 Aromatic proton at C-3”, C-

5”

4. 8.36 s 1H - Aromatic proton at C-1’

5. 8.23-8.26 d 2H 9.0 Aromatic proton at C-2”, C-

6”

6. 7.9-8.17 m 8H - Aromatic protons at C-4, C-

5, C-6, C-3’, C-4’, C-5’,



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

C-8’ and

Ar Ar

O H

7. 7.62-7.66 d 1H 13 Ar Ar

O

H


C-7’

4.57 Compound: J20 (39)

IUPAC Name: N-{3-[3-(4-methoxynaphthalen-1-yl)-3-oxoprop-1-en-1-yl]phenyl}benz

amide



Physical Data:

• State: Solid

• Color: Yellow


Table 4.67: Interpretation of IR spectrum (KBr disk) of J20 (39) [Fig 8.34, Page 192]











9. 973.61 C-H Bending Trans alkene



Table 4.68: Mass Fragmentation of J20 (39) [Fig 8.67 and 8.68, Page 209]

Sr.

No.


Positive ionization

1. 430.3 M+ Na

2. 409 M+2H

3. 408.3 M+H

4. 390.4 M-19

5. 250.3 M-157

NH

H3COO

O

1

23

4

5

6

1'

2'3'

4'

5'

6'

7'

8'

1"2"

3"

4"

5"

6"



Sr.

No.


6. 198.2 M-213

7. 105.2 M-302

Negative ionization

8. 406.3 M-H

Table 4.69: Interpretation of 1H NMR spectrum (CDCl3) of J20 (39) [Fig 8.88, Page 227]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 8.61-8.66 d 1H 15.3 Ar Ar

O H


3. 8.21-8.24 m 2H - Aromatic proton at C-2, C-2’

4. 8.08-8.18 m 2H - Aromatic proton at C-2”, C-6”

5. 7.88-7.95 m 3H - Aromatic protons at C-4, C-8’,

and -CONH Amide proton


7. 7.47-7.62 m 7H -

Aromatic proton at C-6’, C-7’,

C-5, C-3”, C-4”, C-5” and

Ar Ar

O

H


4.58 Compound: PG1 (40)

IUPAC Name: N-{3-[3-(3-Hydroxy-4,5-dimethoxy-phenyl)prop-2-enoyl]phenyl}-4-nitro

benzamide



Physical Data:

• State: Solid

• Color: Yellow


Table 4.70: Interpretation of IR spectrum (KBr disk) of PG1 (40) [Fig 8.35, Page 193]



1. 3430.30 O-H Stretching

N-H Stretching

Phenol

Secondary amide


O

NH

OH

OCH3

OCH3

O2N

O

12

3

4

5

6

1'

2'3'

4'

5'

6'

1"

2"

3"

4"

5"

6"





3. 2929.97 C-H Stretching Alkyl

4. 2862.74 C-H Stretching Ether








Table 4.71: Mass Fragmentation of PG1 (40) [Fig 8.69 and 8.70, Page 210]

Sr.

No.


Positive ionization

1. 471.2 M+ Na

2. 449.2 M+H

3. 435.4 M-15

4. 421.4 M-29

5. 405.2 M-44

6. 391.4 M-59

7. 377.4 M-73

8. 363.2 M-87

9. 283.2 M-165

10. 255.2 M-193

11. 179.0 M-269

12. 153.0 M-295

Negative ionization

13. 447.2 M-H

Table 4.72: Interpretation of 1H NMR spectrum (DMSO) of PG1 (40) [Fig 8.89, Page 228]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 10.78 s (D2O

exchangeable) 1H - -OH

2. 8.38-8.41 m 4H -

Aromatic protons at C-3”, C-

5”, C-2, -CONH Amide

proton

3. 8.21-8.24 d 2H 8.7 Aromatic protons at C-2”, C-

6”

4. 8.10-8.13 d 1H 12 Ar Ar

O H

5. 7.97-8.01 d 1H 12 Ar Ar

O

H



Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

6. 7.71-7.75 d 2H 2.4 Aromatic protons at C-6, C-4


8. 7.20 s 2H - Aromatic protons at C-2’, C-6’

9. 3.84 s 6H - -OCH3

4.59 Compound: PG4 (41)

IUPAC Name: N-{3-[3-(4-Hydroxy-naphthalen-1-yl)prop-2-enoyl]-phenyl}-4-nitrobenz

amide



Physical Data:

• State: Solid

• Color: Yellow

• Melting Point: 198°C

Table 4.73: Interpretation of IR spectrum (KBr disk) of PG4 (41) [Fig 8.36, Page 193]



1. 3310.92 Broad O-H Stretching Phenol


3. 3068.25 C-H Stretching Aromatic, Alkene

4. 1648.35 C=O Stretching α,β-unsaturated Aromatic

Ketone








Table 4.74: Mass Fragmentation of PG4 (41) [Fig 8.71 and 8.72, Page 211]

Sr.

No.


Positive ionization

1. 461.1 M+ Na

2. 439.0 M+H

3. 380.2 M-58

4. 317.2 M-123

5. 270.4 M-170

O

NH

O2N

OOH

1

2

3

4

56

1'

2 '

3 '4'

5'

6'

7 '

8 '

1"

2"

3"

4"

5"

6"



Sr.

No.


6. 241.0 M-197

7. 169.0 M-269

8. 150.1 M-288

Negative ionization

9. 437.2 M-H

Table 4.75: Interpretation of 1H NMR spectrum (DMSO) of PG4 (41) [Fig 8.90, Page 229]

Sr.

No.

Chemical

shift (ppm)

Peak type /

multiplicity

No. of

protons

J value

(Hz)

Proton

1. 9.99 s, D2O

exchangeable 1H - -OH

2. 8.46-8.51 d 1H 15.3 Ar Ar

O H

3. 8.40-8.42 s 1H - -CONH Amide proton

4. 8.13-8.31 m 4H - Aromatic protons at C-2”, C-

5”, C-5’ and C-2

5. 7.56-7.89 m 8H -

Aromatic protons at C-3”, C-

6”, C-4, C-5, C-6, C-6’, C-8’

and

Ar Ar

O

H

6. 7.30-7.34 t 1H 7.5 Aromatic proton at C-7’



Following is the summary of characterization of all reactants used (Table 4.76) and the

intermediates (Table 4.77) and products (Table 4.78) synthesized in the study

Table 4.76: Summary of characterization of all reactants used in the study

Sr.

No.

Reactant (Code) Melting point /

boiling point (°C)

IR (KBr disk, cm-1

)

1. Benzaldehyde (1) 178 3078.70 (C-H Str, aromatic), 2878.68 and

2721.48 (C-H Str, aldehyde), 1691.69 (C=O

Str, aldehyde) , 1617.67 (C-C Str, aromatic)

2. Acetophenone (2) 202 3068.70 (C-H Str, aromatic), 2878.68 (C-H

Str, methyl ketone), 1663.69 (C=O Str,

ketone), 1630.37 (C-C Str, aromatic)

3. 2-Naphthaldehyde (4) 58-60 3062.09 (C-H Str, aromatic), 2828.81 and

2711.48 (C-H Str, aldehyde), 1693.32 (C=O

Str, aldehyde), 1597.20 (C-C Str, aromatic)

4. 3-Hydroxy

acetophenone (5)

94-96 3173.87 (O-H Str, phenol), 2963.58 (C-H Str,

aromatic), 2829.13 (C-H Str, ketone),

1663.96 (C=O Str, ketone), 1578.21 (C-C

Str, aromatic), 1424.71 (C-O-H bend, phenol)



Sr.

No.

Reactant (Code) Melting point /

boiling point (°C)

IR (KBr disk, cm-1

)

5. 3-Aminoaceto-

phenone (7)

96-98 3466.02 (N-H Str, amine), 3219.67 (C-H Str,


1666.59 (C=O Str, ketone), 1629.70 (C-C

Str, aromatic), 1354.77 (C-H bend, ketone)

6. 3,4,5-Trimethoxy

benzaldehyde (13)

76 3010.18 (C-H Str, aromatic), 2942.34 (C-H

Str, ether), 2842.06 and 2753.47 (C-H Str,

aldehyde), 1686.60 (C=O Str, aldehyde),

1233.51 (C-O Str, ether)

7. 4-Hydroxy

acetophenone (15)

100 3166.10 (O-H Str, phenol), 2952.38 (C-H Str,


1663.69 (C=O Str, ketone), 1578.00 (C-C

Str, aromatic), 1424.61 (C-O-H bend, phenol)

8. 1-Hydroxy

naphthalene (18)

96 3223.72 (O-H Str, phenol), 3049.85 (C-H Str,

aromatic), 1633.31 (C-C Str, aromatic),

1362.41 and 1239.54 (C-O-H bend, phenol)

9. 4-(N,N-dimethyl

amino)benzaldehyde

(25)

72-74 2901.31 (C-H Str, aromatic), 2810.32 and

2713.91 (C-H Str, aldehyde), 1596.01 (C-C

Str, aromatic), 1369.91 (C-N Str, amine)

Table 4.77: Summary of characterization of all intermediates used in the study

Sr.

No.

Intermediate

(Code)

M.P /

B.P (°C) %

yield

IR (KBr disk, cm-1

), Mass (EI m/z), 1H NMR

(300 MHz) characterization

1. 3-Benzoylamino

acetophenone (9)

114 90.39 3323.16 (N-H Str, amide), 3109.24 (C-H Str,

aromatic), 3086.83 (C-H Str, ketone), 1667.32

(C=O Str, ketone, amide), 1547.53 (N-H bend,

amide)

2. 3-(4-nitrobenzoyl)

aminoacetophenone

(12)




amide), 1548.67 and 1344.77 (N=O, nitro)

Mass (m/z): 307.2 (M+Na), 285.2 (M+H), 283.1

(M-H)

3. 4-(4-nitrobenzoyl)

oxyacetophenone

(16)

134 98.47 3106.27 (C-H Str, aromatic), 3073.41 (C-H Str,

ketone), 1746.00 (C=O Str, ester), 1683.97 (C=O

Str, ketone), 1525.60 and 1347.14 (N=O Str, nitro),

1179.32 (C-O Str, ester)

4. 1-Methoxy

naphthalene (19)


ether), 1581.37 (C-C Str, aromatic), 1268.3 (C-O

Str, ether)

5. 4-Methoxy-1-

naphthaldehyde

(20)


ether), 2846.54 (C-H Str , aldehyde), 1618.98 (C-C

Str, aromatic), 1248.00 (C-O Str, ether) 1H NMR (DMSO d6): δ 10.18 (s, 1H), 9.23 (d, 1H),

8.29 (d, 1H, J = 8.1 Hz), 8.15 (d, 1H, J = 8.4 Hz),

7.72 (t, 1H, J = 8.1 Hz), 7.75 (t, 1H, J = 8.1 Hz),

7.03 (d, 1H, J = 8.4), 4.10 (s, 3H)

6. 3-(2-naphthoyl)

aminoaceto

phenone (23)




amide)



Sr.

No.

Intermediate

(Code)

M.P /

B.P (°C)

%

yield

IR (KBr disk, cm-1



7. 3-(2-naphthoyl)oxy

acetophenone (27)



Str, ketone), 1263.85 (C-O Str, ester)

8. 3-(3-triflouro

methylbenzoyl)oxy

acetophenone (30)



Str, ketone), 1233.14 (C-O Str, ester), 1122.39 (C-F

Str)

9. 3-(3-triflouro

methylbenzoyl)

aminoacetophenone

(33)


aromatic), 1669.92 (C=O Str, ketone, amide),

1542.97 (N-H bend, amide), 1125.99 (C-F Str)

10. 3-(4-nitrobenzyl)

aminoaceto

phenone (36)

120 75 3342.03 (N-H Str, amine), 2998.87 (C-H Str,



amine), 1518.88 and 1340.13 (N=O Str, nitro)

Table 4.78: Summary of characterization of all products used in the study

Sr.

No.

Product

(Code)

M.P/

B.P (°C) %

yield

IR (KBr disk, cm-1



1. Chalcone

(3)

56 72 IR: 3059.27 (C-H Str, aromatic), 1661.03 (C=O Str, ketone),

1578.67 (C=C Str, alkene)

Mass (m/z): 231.4 (M+Na), 209.4 (M+H), 207 (M-H) 1H NMR (DMSO d6): δ 8.03 (d, 1H, J = 12 Hz), 7.80 (m, 3H),

7.52 (m, 4H), 7.44 (m, 1H), 7.36 (t, 2H, J = 8.1 Hz), 7.07 (m,

1H)

2. D9 (6) 175 45.71 IR: 3385.90 (O-H Str, phenol), 1666.03 (C=O Str, ketone),

1571.23 (C=C Str, alkene), 1272.72 (C-O Str, phenol)

Mass (m/z): 297.5 (M+Na), 275.2 (M+H), 273 (M-H) 1H NMR (DMSO d6): δ 9.85 (s, 1H, D2O exchangeable), 8.10

(d, 1H, J = 8.1 Hz), 7.94 (m, 5H), 7.85 (d, 1H, J = 15.3 Hz),

7.50 (m, 4H), 7.38 (d, 1H, J = 15.3 Hz), 7.08 (d, 1H, J = 8.1)

3. D10 (10) 165 61.98 IR: 3248.27 (C-H Str, amide), 3055.95 (C-H Str, aromatic),

1664.33 (C=O Str, ketone), 1644.86 (C=O Str, amide),

1532.04 (C=C Str, alkene)

Mass (m/z): 400.3 (M+Na), 378.3 (M+H), 376.2 (M-H) 1H NMR (DMSO d6): δ 8.47 (s, 1H), 8.24 (s, 1H), 8.15 (d, 1H,

J = 8.1 Hz), 7.8 (m, 9H), 7.58 (d, 1H, J = 15.6 Hz), 7.4 (m, 6H)

4. DM1

(14)

>250 64.28 IR: 3363.85 (N-H Str, amide), 3076.68 (C-H Str, aromatic),

2835.09 (C-H Str, ether), 1684.56 (C=O Str, ketone), 1646.18

(C=O Str, amide), 1528.20 and 1347.79 (N=O Str, nitro),


Mass (m/z): 485.3 (M+Na), 463.3 (M+H), 461.5 (M-H) 1H NMR (DMSO d6): δ 10.8 (s, 1H), 8.38 (m, 3H), 8.21 (d,

2H, J = 8.7 Hz), 8.11 (d, 1H, J = 9 Hz), 8.00 (d, 1H, J = 7.8

Hz), 7.83 (d, 1H, J = 15.6 Hz), 7.69 (d, 1H, J = 15.3), 7.59 (t,

1H, J = 8.1), 7.24 (s, 2H), 3.86 (s, 9H)

5. DM3

(17)

180 49.38 IR: 3108.23 (C-H Str, aromatic), 2841.67 (C-H Str, ether),

1734.55 (C=O Str, ester), 1669.44 (C=O Str, ketone), 1585.24

(C=C Str, alkene), 1507.90, 1350.19 (N=O Str, nitro), 1262.05

(C-O Str, ether)

Mass (m/z): 486.2 (M+Na), 464.3 (M+H), 462 (M-H)



Sr.

No.

Product

(Code)

M.P/

B.P (°C)

%

yield

IR (KBr disk, cm-1


(300 MHz) characterization 1H NMR (DMSO d6): δ 8.4 (m, 4H), 7.96 (d, 1H, J = 7.8),

7.87 (m, 1H), 7.73 (d, 1H, J = 15.9 Hz), 7.59 (d, 1H, J = 8.1

Hz), 7.46 (d, 1H, J = 8.1Hz), 7.35 (d, 1H, J = 15.9), 6.87 (s,

2H), 3.91 (s, 9H)

6. DM5

(21)

230 63.15 IR: 3411.02 (N-H Str, amide), 2935.57 (C-H Str, aromatic),



1518.88 (C=C Str, alkene) 1254.48 (C-O Str, ether)

Mass (m/z): 475.3 (M+Na), 453.3 (M+H), 451.3 (M-H) 1H NMR (DMSO d6): δ 10.2 (s, 1H), 8.56 (s, 1H), 8.49 (d, 1H,

J = 8.1 Hz), 8.37 (d, 2H, J = 8.7), 8.22 (m, 5H), 8.12 (d, 1H, J

= 8.1 Hz), 7.99 (d, 1H, J = 8.1 Hz), 7.84 (d, 1H, J = 15.3 Hz),

7.67 (t, 1H, J = 8.1), 7.58 (m, 2H), 7.13 (d, 1H, J = 8.1), 4.06

(s, 3H)

7. DM7

(24)



(C=O Str, amide), 1528.63 (C=O Str, alkene), 1261.22 (C-O

Str, ether)

Mass (m/z): 490.3 (M+Na), 468.4 (M+H), 466.3 (M-H) 1H NMR (DMSO d6): δ 8.50 (s, 1H), 8.43 (s, 1H), 7.99 (m,

2H), 7.86 (m, 4H), 7.78 (d, 1H, J = 7.8 Hz), 7.65 (d, 1H, J =

15.9 Hz), 7.51 (m, 3H), 7.36 (d, 1H, J = 15.9 Hz), 6.78 (s, 2H),

3.88 (s, 9H)

8. DM8

(26)


1650.34 (C=O Str, ketone, amide), 1320.96 (C-N Str, amine)

Mass (m/z): 443.0 (M+Na), 421.1 (M+H), 419.5 (M-H) 1H NMR (DMSO d6): δ 8.8 (s, 1H), 8.42 (s, 1H), 8.24 (m, 2H),

7.98 (m, 6H), 7.57 (m, 5H), 7.42 (d, 1H, J = 15.3 Hz), 6.98 (m,

2H), 3.2 (s, 6H)

9. E3 (28) 185 65.70 IR: 3050.34 (C-H Str, aromatic), 1726.81 (C=O Str, ester),

1658.74 (C=O Str, ketone), 1600.28 (C=C Str, alkene),


Mass (m/z): 451.4 (M+Na), 429.3 (M+H), 427.0 (M-H) 1H NMR (DMSO d6): δ 8.9 (s, 1H), 8.37 (s, 1H), 8.07 (m,

8H,), 7.98 (m, 3H), 7.85 (s, 1H, J = 15.3 Hz), 7.65 (m, 4H),

7.56 (t, 2H)

10. E5 (31) 180 69.93 IR: 3025.21 (C-H Str, aromatic), 1734.31 (C=O Str, ester),

1659.95 (C=O Str, ketone), 1602.27 (C=C Str, alkene),


Mass (m/z): 469.5 (M+Na), 447.2 (M+H), 445.4 (M-H) 1H NMR (DMSO d6): δ 8.5 (s, 1H), 8.43 (d, 1H, J = 7.8), 7.85

(m, 8H), 7.79 (d, 1H, J = 13.2 Hz), 7.47 (m, 6H)

11. F3 (32) >250 91.57 IR: 3221.29 (N-H Str, amide), 3055.83 (C-H Str, aromatic),


1586.08 (N-H bend, amide)

Mass (m/z): 450.3 (M+Na), 428.3 (M+H), 426.3 (M-H) 1H NMR (DMSO d6): δ 10.69 (s, 1H), 8.65 (s, 1H), 8.53 (s,

1H), 8.36 (s, 1H), 8.19 (d, 1H, J = 9.6 Hz), 7.95 (m, 10H), 7.87

(d, 1H, J = 15.9), 7.57 (m, 5H)

12. F5 (34) 170 70.17 IR: 3247.44 (N-H Str, amide), 3054.70 (C-H Str, aromatic),


1548.29 (C=C Str, alkene), 1054.31 (C-F Str)



Sr.

No.

Product

(Code)

M.P/

B.P (°C)

%

yield

IR (KBr disk, cm-1



Mass (m/z): 468.3 (M+Na), 446.3 (M+H), 444.3 (M-H) 1H NMR (DMSO d6): δ 10.71 (s, 1H), 8.46 (s, 1H), 8.36 (s,

2H), 8.30 (d, 1H, J = 8.1 Hz), 8.10 (m, 2H), 7.9 (m, 7H), 7.78

(t, 1H, J = 7.5), 7.62 (s, J = 12 Hz), 7.57 (m, 2H)

13. F7 (37) 192 61.50 IR: 3342.41 (N-H Str, amine), 3055.97 (C-H Str, aromatic),

1665.26 (C=O Str, ketone), 1600.67 (C-C Str, aromatic),

1583.65 (C=C Str, alkene), 1518.54 and 1342.76 (N=O Str,

nitro)

Mass (m/z): 431 (M+Na), 409 (M+H), 407 (M-H) 1H NMR (DMSO d6): δ 8.37 (m, 2H), 7.9 (m, 2H), 7.86 (d,

1H, J = 15.2 Hz), 7.55 (m, 10H), 7.20 (m, 2H), 4.25 (s, 2H),

3.8 (s, 1H)

14. G10 (38) 235 67.11 IR: 3391.95 (N-H Str, amide), 2924.36 (C-H Str, aromatic),


1584.44 and 1347.19 (N=O Str, nitro), 1526.82 (N-H Str,

amide)

Mass (m/z): 445.3 (M+Na), 423 (M+H), 421.2 (M-H) 1H NMR (DMSO d6): δ 10.88 (s, 1H), 8.5 (s, 1H), 8.37 (d, 2H,

J = 9.0 Hz), 8.36 (s, 1H), 8.23 (d, 2H, J = 9.0 Hz), 7.9 (m, 8H),

7.62 (d, 1H, J = 13), 7.57 (m, 2H)

15. J20 (39) 118 59.63 IR: 3347.80 (N-H Str, amide), 3071.01 (C-H Str, aromatic),


(C=O Str, amide), 1557.99 (C=C Str, alkene), 1254.71 (C-O

Str, ether)

Mass (m/z): 430.3 (M+Na), 408.3 (M+H), 406.3 (M-H) 1H NMR (DMSO d6): δ 8.61 (d, 1H, J = 15.3), 8.33 (d, 1H, J =

8.1), 8.21 (m, 2H), 8.08 (m, 2H), 7.88 (m, 3H), 7.83 (d, 1H, J

= 8.1 Hz), 7.47 (m, 7H), 6.88 (d, 1H, J = 8.1 Hz)

16. PG1 (40) 210 31.25 IR: 3430.30 (N-H Str, phenol), 2963.58 (C-H Str, aromatic),




Mass (m/z): 471.2 (M+Na), 449.2 (M+H), 447.2 (M-H) 1H NMR (DMSO d6): δ 10.78 (s, 1H, D2O exchangeable),

8.38 (m, 4H), 8.21 (d, 2H, J = 8.7 Hz), 8.10 (d, 1H, J = 12 Hz),

7.97 (d, 1H, J = 12 Hz), 7.71 (d, 2H, J = 2.4 Hz), 7.57 (t, 1H, J

= 8.1), 7.20 (s, 2H), 3.84 (s, 6H)

17. PG4 (41) 198 41.66 IR: 3310.92 (O-H Str, phenol), 3200.16 (N-H Str, amide),

3068.25 (C-H Str, aromatic), 1648.35 (C=O Str, ketone),

1641.06 (C=O Str, amide), 1580.41 and 1388.94 (N=O Str,

nitro), 1179.321254.35 (C-O Str, phenol)

Mass (m/z): 461.1 (M+Na), 439 (M+H), 437.2 (M-H) 1H NMR (DMSO d6): δ 9.99 (s, 1H, D2O exchangeable), 8.46

(d, 1H, J = 15.3 Hz), 8.40 (s, 1H), 8.13 (m, 4H), 7.56 (m, 8H),

7.30 (t, 1H, J = 7.5 Hz), 7.11 (d, 1H, J = 8.4), 6.59 (d, 1H, J =

8.4 Hz)



D. PHARMACOLOGICAL EVALUATION OF SYNTHESIZED ANALOGUES

Synthesized analogues were screened to determine angiogenesis inhibition activity.

• Sixteen synthesized compounds were screened for anti-angiogenic activity by

Chorioallantoic membrane (CAM) assay.

• Minocyclin hydrochloride was used as the standard and dimethyl sulphoxide (DMSO)

was used as the vehicle for the assay.

4.60 Experimental design

4 days old white Leghorn fertilized chicken eggs were procured from central poultry

development organization, Govt. of India, Mumbai.

They were cleaned with soap solution and surface sterilized using 70 % alcohol. They were

kept in a humidified incubator at 37°C +/- 0.3°C.

The eggs were maintained in vertical position. On day 5, the eggs were again cleaned with

70 % alcohol. The upper surface of the egg was pierced with 18 gauge needle.

10 µl of vehicle (negative control), standard (Positive control) or test compound solutions of

suitable concentrations (in DMSO) were injected into the cavity through the opening.

The openings were sealed with adhesive tape and the eggs were re-incubated in humidified

incubator at 37°C.

On day 12, the CAMs were harvested and fixed in 10% formalin in Phosphate Buffered

Saline (PBS) pH 7.4 solution. The CAM’s were stained with Hematoxylin and Eosin (H&E)

using standard protocol. For every concentration of solutions of test compounds, positive

and negative controls, 6 eggs were used.

4.61 Capturing of CAM Images for analysis of anti-angiogenic effect

The images of CAMs were captured on a white background using Sony Cybershot DSC

W55 7.2 megapixels digital camera with 5x optical zoom, resolution of 640 × 480 pixels.

The images were also captured using Olympus microscope with 50x magnification and

dissection microscope with 20x magnification.

4.62 Image processing and Quantification of angiogenesis

The images of CAMs were analyzed using image analysis software AngioQuant v 1.33 (a

MATLAB based software tool for quantification of angiogenesis), ImageJ (NIH Software)

and SPIP software for the analysis of blood vessels.



4.63 Analysis of blood vessel pattern formation

Captured Digital images of CAM were analyzed for various analyzed for various distorted

patterns of blood vessels and these images were further used for ranking of blood vessel

patterns from 0 to ++++ based on observations like, disorganizations, bending, sharp turns

and disruptions in vessel, where 0 represents the branching pattern observed in controls and

++++ representing the most severe disruption of pattern (Table 4.79).

Table 4.79: Criteria for ranking of blood vessel pattern

Rank Criteria

Central vessel Branching pattern Collateral vessel

0

λ shape Normal tree branch pattern,

well organized pattern Vessels branch out from

lower side of membrane

Prominent central

vessel

Central vessel branches

from upper to lower side

evenly

Slight development of

collateral vessels towards

end of membrane The vessel begins from

upper side of

membrane

No turns, bends seen in

branches of vessels Minimum branching of

vessels

Long central vessel

ends at the lower side

of membrane

Primary and secondary

vessels are visible No disorganization in

collateral vessel pattern

-- No disorganizations in

branching pattern --

-- No cross-overs or wavy

vessels --

+ λ shape sustained Slight disorganizations in

branching pattern

Collateral development -

individual branching

pattern seen

Central vessel visible Some cross-over of blood

vessels Significant vessels

development seen

-- Uneven branching of

central vessels --

++ λ shape sustained Bent vessels Major collateral vessel

development

Central vessel visible Sharp turns seen in vessels

- vessel changes the

direction abruptly

The branching of

collateral vessel may

reach upper side of

membrane

Length of central vessel

reduced - does not

reach end of membrane

Major cross-over seen in

blood vessel patterns with

localized low and high

branching points

--

+++ λ shape distorted or

missing Long parallel vessels

Major collateral vessel

development

Central vessel visible Central vessel with upper

side with less or no

branching

Extensive multiple

collateral vessels

development

Length of central vessel

reduced - does not

Lower side of central vessel

shows branching Branching of collateral

vessels till upper part of



Rank Criteria

Central vessel Branching pattern Collateral vessel

reach end of membrane membrane

Multiple vessels from

upper side of

membrane instead of

one central vessel

branching downwards

i.e. Appearance of long

vessel with branching more

at lower side compared to

upper part of CAM

Disorganized collateral

vessels

-- Major disorganized blood

vessels --

++++ λ shape may be missing Random orientations of

blood vessels Some collateral

development

Distorted central vessel Wavy blood vessels Extreme disorganization

or distortions of collateral

vessels

Short central vessel Severe disruption of blood

vessel pattern Less branching of

collateral vessels

central vessel may be

missing No / less branching of

blood vessels Remarkable reduction in

size of CAM

Average anti-angiogenic score for observed blood vessel pattern was calculated as follows -

Where – Score 1: +, Score 2: ++, Score 3: +++, Score 4: ++++

Based on the ranks obtained, average anti-angiogenic score was calculated which is

interpreted as follows-

Anti-angiogenic score

0-1: No anti-angiogenic effect

1-2: weak anti-angiogenic effect

2-3: good anti-angiogenic effect

>3: very good anti-angiogenic effect

4.64 Quantification of total area of CAM

To determine CAM area, each CAM was placed in a Petri plate and the longest and shortest

lengths were measured using a Vernier caliper to a precision of 0.02 mm. Data were

recorded in Excel spreadsheets, and CAM areas in mm2 were calculated using the formula:

CAM area = Longest length

× Longest width

× 3.14 2 2

Average =

score

No of eggs (score 1) × 1

+


+


+


Total No. of eggs

(score 1 -4)

Total No. of eggs

(score 1 -4)

Total No. of eggs

(score 1 -4)

Total No. of eggs

(score 1 -4)



Mean standard deviation, standard error of mean and % relative standard deviations were

calculated for vehicle control, standard and test compounds.

A graph of average area ± SEM in mm2 vs. concentrations of vehicle / standard / test

compound in µM was plotted.

Effect of various concentrations of DMSO (0.001%, 0.01%, 0.1% & 1%) as vehicle on CAM

area growth was determined by comparing their average CAM area with the one obtained

using water.

Further, the means of CAM area of vehicle controls, standard and test compounds were

compared to determine the effect of standard and test compounds on CAM growth.

For each experiment, data was analyzed using one way analysis of variance (ANOVA) for

checking significance.

4.65 Statistical analysis

Each value was expressed as a mean ± Standard error of mean (S.E.M.) The statistical

significance of results was determined by one-way analysis of variance (ANOVA). P value

<0.05 was considered statistically significant.



E. QUANTITATIVE STRUCTURE ACTIVITY RELATIONSHIP (QSAR) STUDY

OF SYNTHESIZED MOLECULES BASED ON PHARMACOLOGICAL

EVALUATION

QSAR analysis was performed to establish the relationship between anti-angiogenic activity

of the synthesized molecules and their molecular descriptors.

4.66 Softwares used in study

• Molecular modeling softwares used for the QSAR study were -

o Schrödinger software [Maestro®

7.5 (Graphical user interface), USA] on Red Hat

Linux Enterprise platform.

� Ligprep module

� QikProp module

� Strike module

• 2D and 3D structures of molecules were built and cleaned-up in CS Chem Draw Ultra

7.01, Cambridge Soft Corporation USA

4.67 QSAR study of synthesized molecules based on angiogenesis inhibition activity

4.67.1 Dataset of compounds and parameters

Test compounds were divided into training set of thirteen compounds and test set of four

compounds randomly.

To derive QSAR model, descriptors of physicochemical properties were used as independent

variables and the activity parameter, log (% anti-angiogenic score at 10µM concentration)

was used as dependant variable.

4.67.2 Ligand preparation

The structures of synthesized angiogenesis inhibitors were imported to Maestro. They were

processed using Ligprep module and minimized with Optimized Potentials for Liquid

Simulations (OPLS_2005) force field.

4.67.3 Descriptors of compounds

To obtain the quantitative effect of structural parameters of the substituted 1,3-

diarylpropenone derivatives on their anti-angiogenic activity, QSAR analysis with



physicochemical descriptors was performed. Thirty physicochemical descriptors were

calculated using QikProp module of Schrödinger software.

4.67.4 Statistical Analysis

All the statistical analysis was carried out in Strike module of Schrodinger

4.67.4.1 Correlation Analysis

Correlation analysis of biological activity and physicochemical parameters was carried out.

Inter-correlated parameters were eliminated stepwise depending on individual correlation

with biological activity. Possible combinations of parameters were considered for multiple

regression analysis. For final statistical analysis, a set of four descriptors was selected on the

basis of correlation matrix, descriptor significance and training set size.

4.67.4.2 Multiple Regression Analysis

The QSAR model was generated by multiple linear regression (MLR) analysis using Strike

module. The statistical quality of regression equations was justified by parameters like,

correlation coefficient (R), variance ration (F), standard deviation (SD), F-test – Fisher test

for significance of the equation and Standard error of estimate (s).

All the final equations had significant regression coefficient, intercept and variance ratio (F).

Use of more than one variable in multivariate equation was justified by auto-correlation

study. Best QSAR model was selected on the basis of squared correlation coefficient (r2),

standard deviation (SD), Fisher’s value (F) and p as the statistical parameters.

4.67.4.3 Cross Validation

The predictive power of the generated QSAR model was validated by Leave One Out (LOO)

cross validation method with computation of Q2

and test set prediction.

4. 67.5 Interpretations of QSAR study

Using experimental predicted values of log (% anti-angiogenic score) for training and test set

of compounds, residual values were calculated.

Graph of residual log (% anti-angiogenic score) against experimental (% anti-angiogenic

score) was plotted.

experimental work - shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/7005/20/10_chapter...

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