r

1. •Professor Dr. khairia Mohammed Youssef, Dept. of
Pharm. Chemistry, Faculty of Pharmaceutical Sciences
and Pharmaceutical Industries, Future University,P.O.
BOX 12311, Cairo, Egypt
•khairiayoussef@yahoo.com
•kyoussef@fue.edu.eg
Targeted Drug Delivery System (TDDS):
Encapsulating Newly Synthesized Anti-cancer
Compounds-conjugated gold nanoparticles

2. • 1- Introduction
• 2- Synthetic Methods of Curcumin and Curcumin
analogs.
• 3-In vitro Assessment of Curcumin and Curcumin
Analogs as Anti-Oxidant and Anti-Cancer Agents.
• 4-Mechanisms of Action of Curcumin and
Curcumin analogs as Anti-oxidant and Anti-
Cancer Agents.
• 5- Mechanism of Cytotoxic Tubulin Polimerization
Activities.
• 6- In vivo Assessment of Curcumin as Anti-
Cancer.
– Preclinial Safety Evaluation in Mice and Rats.
– Chemoprotective and Chemopreventive effects
in DMH-Induced Colon Cancer in Albino Rats
Model.
7- Conjugation with gold nanoparticles

3. Introduction:
Cancer and Curcumin as anticancer
• Cancer is perhaps the most progressive and
devastating disease posing a threat of mortality to
the entire world despite significant advances in
medical technology for its diagnosis and treatment.
• Recently, considerable attention has been focused on
identifying naturally occurring chemopreventives.
• Wide arrays of phenolic substances, particularly those
present in dietary and medicinal plants, have been
reported to possess substantial anticarcinogenic and
antimutagenic effects

4. Curcumin
diferuloylmethane, 1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione
the major constituent of turmeric powder extracted from the rhizome of the plant
curcuma longa

5. Biological Properties of Curcumin and
its Derivatives
• 1. Antioxidant
• 2. Anti-inflammatory
• 3. Antibacterial, Antifungal and Antiparasitic
• 4. Antiviral
• 5. Antihistaminic
• 6. Treatment of Skin Diseases
• 7. Chemopreventive Effects
• 8. Anticancer
• 9. Treatment of Alzeheimer

6. Synthetic Methods for the Preparation
of Curcumin and Curcumin derivatives
CHO
RO
RO
O O
OR
OH
HO HO
NaOH/EtOH, NaOEt/EtOH
or c.HCl
XXX
O O
+
1
1
2
1
In an attempt to prepare curcumin and curcumin analogs, search of the literature
demonstrated that chalcones, Claisen-Schmidt condensation in general, are known
to be prepared via the classic enolate condensation reaction which is an acyl
addition reaction of a nucleophilic enolate to an electrophilic carbonyl carbon
In 1973, Roughley and Whiting reported the instability of curcumin in alkaline
medium and explained its alkaline degradation as shown in the following equations:

7. alkaline degradation of Curcumin
H3CO
O O
OCH3
OH
HO
i
HO
COOH
H3CO
HO
H3CO
O
+
iv
i
CO2 (BaCO3)
H3CO
HO
CHO
+ CH3COCH3
iii ii
CHI3
H3CO
HO
COOH
iv
CO2 (BaCO3)
Ferulic acid
Feruloylmethane
acetone
vanillin
Vanillic acid
iodoform
Roughley, P.J. and Whiting, D.A.; J. Chem. Soc. Trans I (1973),
2379-2388.

8. Pabon Procedure for the Synthesis
of Curcumin and its Analogs
R1O
HO
OR1
OH
O O
R1O
HO
CHO
+
O O
/ B2O3
(BuO)3B/ n-BuNH2
2
• Pabon, H.J.J; Rec. Tr. Chim. Pays-Bas (1964), 83: 379-386.
• Synthesis of Curcumin Analogues as Potential Antioxidant, Cancer Chemopreventive Agents. Khairia M.
Youssef, Magda A. El-Sherbeny, Faiza S. El-Shafie, Hassan A. Farag, Omar A. Al-Deeb, Sit Albanat A.
Awadalla. Arch. Pharm. Pharm. Med. Chem. 2004, 337, 42-54.
• Synthesis and Antitumor Activity of Some Curcumin Analogs. Part II. Khairia M. Youssef, Magda A. El-
Sherbeny; Arch. Pharm. Chem. Life Sci., 338, 181-189, 2005.
• Synthesis of bioconjugate analogs of curcumin as potent antitumor agents. Khairia M. Youssef, Omaima A.
Abu-Alwafa, Reem I. Al-Wably. Med Chem Res (2012) 21:874–890.
the acetylacetone-boric anhydride complex method was adopted successfully for the
preparation of Hundreds of Curcumin and Curcumin Derivatives.

9. H3CO
O O
OCH3
OH
HO
CH3
H3C
O
O O
B
O
O O
CH3
H3C
H3C CH3
O
H
HO
OCH3
+ B2O3 + BO2
-
+ H2O
(A)
n-BuNH2
O
B
O
O O
CH=CH
HC=HC
HC=HC CH=CH
OH
OCH3
HO
OCH3 OCH3
O+
H
H3CO
H+
O
(B)
2 (C)
HCl
4
Cl-
Cl-
2

10. Synthetic Methods for the Preparation
of Curcumin Analogs
• The Biological Study of Curcumin reported
the instability of curcumin at a pH above 6.5
which is attributed to the active methylene
group.
• Omitting the active methylene group and one
carbonyl group may lead to potent
antioxidative compounds
O
O
OH
OCH3
HO
OCH3

11. Synthesis of Curcumin Analogs
CHO
R'O
OCH3
N
O
R
N
O
OR'
OR''
R'O
OR'' R
+
R = H, Me, Et, Propyl, acetyl
R' = H, Me, Et
R''= H, Me, Et, alkyl derv.
3,5-Bis(substituted benzylidene)-N-alkyl-4-piperidones was obtained through
condensation of the appropriate 1-alkyl-4-piperidone with the appropriate aldehyde
under acidic condition.

12. Prepared Compounds
N
O
OH
OC2H5
HO
OC2H5 CH3
N
O
OH
OC2H5
HO
OC2H5 C2H5
N
O
OH
OCH3
HO
OCH3 CH3
• Synthesis of Curcumin Analogues as Potential Antioxidant, Cancer Chemopreventive Agents. Khairia M.
Youssef, Magda A. El-Sherbeny, Faiza S. El-Shafie, Hassan A. Farag, Omar A. Al-Deeb, Sit Albanat A.
Awadalla. Arch. Pharm. Pharm. Med. Chem. 2004, 337, 42-54.
• Synthesis and Antitumor Activity of Some Curcumin Analogs. Part II. Khairia M. Youssef, Magda A. El-
Sherbeny; Arch. Pharm. Chem. Life Sci., 338, 181-189, 2005.
• Synthesis and in vitro antioxidant activity of some new fused pyridine analogs. Mohamed A. Al-Omar, Khairia
M. Youssef, Magda A. El-Sherbeny, Sit Albanat A. Awadalla, Hussein, I. El-Subbagh; Arch. Pharm. Pharm.
Chem. Life Sci. 2005, 338, 181-189.
N
CH3
O
OH
OCH3
HO
OCH3
O
O
OH
OCH3
HO
OCH3

13. (a) (b)
(1)
HO OH
O O
H3CO OCH3
HO OH
OH O
H3CO OCH3
Curcumin and curcumin derivative have a unique conjugated
structures which show a typical radical trapping ability as a chain-
breaking antioxidant
Curcumin as Anti-Oxidant Agents

14. Mechanism of Antioxidant Activity
of Curcumin
O O
O O
H3CO OCH3
.
.
.
AH: is the phenolic antioxidant.
A●: is the antioxidant radical.
X●: is another radical species or the same as A●.
During Metabolism, Biomolecules
produce reactive peroxyl radicals
during their oxidation, which may act
as X● and couple with the antioxidant
radical A● in the second step of the
antioxidation process

15. 1- In vitro Chromatographic Determination
of Free Radical Scavenging Activity of
Curcumin Analogs
a: Diphenylpicrylhydrazyl Free Radical Test
• Diphenylpicrylhydrazyl absorbs strongly at 515 nm. The
reaction is carried out under pseudo-first-order conditions using
excess of the tested compound compared to the reagent and
monitoring the mixture for the decrease of absorption at 515
nm.
• Form the linear regression line of the absorbency against time,
the pseudo-first-order rate constant (Kapp) and the
corresponding half-life time (t1/2) of the reaction was calculated.

16. b- Chemiluminescence Measurement Scavenging
Activity of Curcumin Analogs of Free Radical oxygen
Produced by Peripheral multinuclear Neutrophil
Cells (PMNs)
• All of the synthesized compounds were tested for their ability
to scavenge oxygen free radical produced by peripheral
multinuclear neutrophil cells (PMNs) collected from apparently
healthy blood donors.
• Phorpol-12-myristate-13-acetate (PMA), in a final
concentration of 2ng/ml was added to PMNs (5x105cells/ml) to
stimulate respiratory burst which was magnified by luminol
(10-4M) to be able to be measured by the LKB luminometer.
• 100µg of synthesized drugs were added to detect its effect on
the amount of oxygen radical liberated and the percentage of
inhibition was calculated.

17. Conclusion
• The Anti-oxidant screening of most of Curcumin Analogs reveals that the
results go hand by hand with the in vitro Anti-cancer effects results which
performed at The National Cancer Institue (NCI).
• The anti-oxidant and anti-cancer effects of these compounds depend
mainly on the stabilization of the formed phenoxy free radical. The p-
hydroxy phenyl moiety is very essential to produce the phenoxy free
radical which is responsible for free radical scavenging effect.
• o-Substitution by electron–donating group like o-methoxy group, increases
stability of phenoxy free radical and hence increasing both free scavenging
and antitumor effects.
• o-Substitution by ethoxy group rather than methoxy, increases stability of
phenoxy free radical and thus increasing both free scavenging and
antitumor effects.
• Increasing the alkyl group chain from methyl to ethyl on the N in the series
of substituted N-alkyl piperidones, increases the activity toward both free
radical scavenging and antitumor effects which may be attributed to
increased positive inductive effect and / or increased lipophilicity of the
new compounds.
• Extension of conjugation, increases stabilization of phenoxy free radical.

18. Chemopreventive Properties of Curcumin and
Curcumin Analogs
Mechanism of Action
• Stimulation of phase I and phase II detox
systems
• Inhibition of COX-1 and COX-2 enzymes
• Stimulation of glutathione-S-transferase
• Interference with cell growth by inhibition of
protein kinases
• Neutralization of carcinogenic free radicals
• Curcumin significantly inhibits the activity of the
isoenzymes of cytochrome P-450 involved in the
metabolism of some carcinogens
• It inhibits SK-Hep-1 hepatocellular carcinoma cell
invasion in vitro and suppresses matrix
metalloproteinase-9 secretion
• Cytotoxic Tubulin Polymerization
Activities

19. Invitro Assessment as Cytotoxic Targeting
Tubulin Polymerization Activity
*Mary Ann Jordan & Leslie Wilson, “Microtubules as a target for anticancer drugs”. Nat. Rev. Cancer, (2004), 4:
253-265.
Microtubules represent one of the fiber systems of the eukaryotic
cytoskeleton. They are essential for a wide variety of cellular functions,
notably: 1- cell motility, 2-transport, 3-cell shape, 4-polarity and 5-mitosis.
Microtubules consist of a core cylinder built from heterodimers of α and β
tubulin monomers.

20. Microtubules binding sites
There are three different target sites on the tubulin
heterodimer:the vinca alkaloid, the colchicine, and
the paclitaxel binding sites.
a. Vinblastine bound to high-
affinity sites at the
microtubule plus end suffice
to suppress microtubule
dynamics.
b. Colchicine forms complexes
with tubulin dimers and
copolymerizes into the
microtubule lattice,
suppressing microtubule
dynamics.
c. A microtubule cut away to
show the interior surface is
shown. Paclitaxel binds
along the interior surface of
the microtubule,
suppressing its dynamics.

21. Tubulin Inhibition Study of
Curcumin Analogs
*Chakrabarti R, Rawat PS, Cooke BM, Coppel RL, Patankar S, “Cellular Effects of Curcumin on Plasmodium falciparum
Include Disruption of Microtubules”. PLoS ONE, (2013), 8 (3): e57302.
curcumin diketo form at the interface of Panel A shows all the predicted
bound poses, mostly at the interface of the dimer.
Panel B shows the most probable binding pose according to Autodock (Rank
1) with the curcumin diketo form at the interface of alpha and beta tubulin
monomers.
Panel C shows binding sites of colchicine (purple), paclitaxel (red) and
vinblastine (brown) on parasite tubulin dimer. Curcumin was proved to have
inhibition activity on tubulin inside P. falciparum cells with IC50 = 5 µM.
tubulin dimer of
P. falciparum
tubulin is
represented as
dimer of alpha
(yellow) and beta
(green) subunit.

22. SAR Performed on Novel Curcumin
Analogs
Docking Study Using Discovery
Studio 2.5 and MOE Programs
R' N
O
R'
O
Auxiliary
binding site
B
Groups capable of Van
der Waal bonding
Atoms capable of
hydrogen bonding
1, 5-diaryl-3-oxo-1,4-
pentadienyl pharmacophore
[Reacts at the primary binding
site A of colchicine]
The group on N atom could lead to increases in cytotoxic potencies due to either additional binding with cellular
constituents at an auxiliary binding site (site B) or by facilitating the interaction of the cytotoxin at site A.
Newly Designed and Synthesized Curcumin Analogs with in vitro Cytotoxicity and Tubulin Polymerization
Activity. Iten M. Fawzy, Khairia M. Youssef, Nasser S. M. Ismail, Joachim Gullbo and Khaled A. M. Abouzid, Chemical Biology &
Drug Design. Early View, Article first published online: 25 NOV 2014.
European Patent, PCT/EG 2013/000028.

23. OH
CH3
N
O
CH3
HO
CH3
O
O
OH
OCH2CH3
HO
OCH2CH3
O
O
OH
OCH3
HO
OCH3
Curcumin
Ethyl Curcumin
Curcumin Analog
Curcumin (1) and curcumin analogs (2,3) were synthesized and the preclinical safety
evaluation in mice and rats were done.
Invivo Assessment as Anticancer Agents

25. Preclinical Safety Evaluation in Mice and
Rats
Acute Toxicity (LD50) of tested
compounds procedure
Doses which used in the assessing of
chemopreventive, antitumor effects
Compound 1 (curcumin) 2 3
Dose 0.05mg/gm/orally/day 0.12mg/gm/orally/day 0.16mg/gm/ora
lly/day
Litchfield JT, Wilconsxon F. J. Pharmcol. Exp. Ther. 1949, 96, 99-113.
Pilot experiments were performed by oral administration of increasing doses of tested
compounds into different groups of mice (each consisted of 5 mice). Doses which
produce 0 % and 100 % mortality were determined.

26. Blood Pressure and Electrocardigrahic
Recording
Test Control Curcumin 2 3
B.P. Mean 109 108.4 109.9 103.9*
± SEM ± 0.573 ± 0.57 ± 0.575 ± 0.546
Three groups of virgin Sprague-Dawley female rats, each consisted of 10 rats: Group A
administered the calculated dose of compounds 1, 2 and 3 daily for two weeks. Group B
administered the calculated dose of compounds 1, 2 and 3 daily for four weeks.

27. Results of Blood Pressure and
Electrocardigrahic Recording
• These tests show that there is significant
reduction in systolic B.P. only with
pretreatment with compound 3 for 14
days P.O. administration at P , 0.05 while
there is no changes in E.C.G. or heart
rate.

28. Histopathological Study
• Different organs sections of untreated and
treated animals e.g. mammary gland,
heart, kidney, liver, spleen and colon were
stained with routine hematoxylin and
eosin (HX and E) to be examined under
light microscope (Fig. 2-7)

29. Histopathological Results
Fig. (1) Light microscope image of
the Mammary gland stained with
H&E (X20)
Group 1: Control (untreated)
Fig. (2) Light microscope image of
the Heart stained with H&E (X10)
Fig. (3) Light microscope image of
the Kidney stained with H&E (X40)
Fig. (4) Light microscope image of
the Liver stained with H&E (X20)
Fig. (5) Light microscope image of
the Spleen stained with H&E (X20)

30. Fig. (6) Light microscope image of
the Lung stained with H&E (X20)
Fig. (7) Light microscope image
ofthe Colon stained with H&E (X20)

31. Group 2 : Compound 1 (Curcumin) After Four Weeks
Fig. (8) Light microscope image of
the Mammary gland stained with
H&E (X20)
Fig. (9) Light microscope image of
the Heart stained with H&E (X10)
Fig. (10) Light microscope image of
the Kidney stained with H&E (X40)
Fig. (11) Light microscope image of
the Liver stained with H&E (X20)
Fig. (12) Light microscope image of
the Spleen stained with H&E (X20)

32. Fig. (14) Light microscope image of the Colon
stained with H&E (X20) showing normal
structure
Fig. (13) Light microscope image of
the Lung stained with H&E (X20)

33. Group 3: Compound 2 (Ethyl Curcumin)
Fig. (15) Light microscope image of
the Mammary gland stained with
H&E (X20)
Fig. (16) Light microscope image of
the Heart stained with H&E (X10)
Fig. (17) Light microscope image of
the Kidney stained with H&E (X40)
Fig. (18) Light microscope image of
the Liver stained with H&E (X20)
Fig. (19) Light microscope image of
the Spleen stained with H&E (X20)

34. Fig. (21) Light microscope image of
the Colon stained with H&E (X10)
showing normal structure
Fig. (20) Light microscope image of the
Lung stained with H&E(X20) showing
thickening of the interestial space

35. Group 4: Compound 3
Fig. (23) Light microscope image
of the Heart stained with H&E
(X10)
Fig. (24) Light microscope image
of the Kidney stained with H&E
(X40)
Fig. (25) Light microscope image
of the Liver stained with H&E (X20)
Fig. (22) Light microscope image
of the Mammary gland stained
with H&E (X20)
Fig. (26) Light microscope image of
the Spleen stained with H&E (X20)

36. Fig. (27) Light microscope image of the Lung stained
with H&E (X20) showing in
(a) Accumulation of fluids in the alveoli (b) Accumulation of lymphocytes and RBC
in the alveolar space

37. Fig. (28) Light microscope image
of the Colon stained with H&E
showing an increase in the goblet
cells in the base of the crypts (arrows)
(a) X10
(b) X40

38. Conclusion
 compound 1 (curcumin) and compound 2 (ethyl curcumin
showed normal structure after 4 weeks treatment identical
of that in 2 weeks .
Compound 3 shows some intraalveolar lung haemorhage
after four weeks.

39. Chemoprotective Effect of
Curcumin and its Analogs in
DMH-Induced Colon Cancer in
Albino Rats Model.
Bird RP. Role of aberrant crypt foci in understanding the pathogenesis
of colon cancer .Cancer Letters 1995; 93 : 55-71.

40. Histological evaluation and Aberent
Cript Focy assay
• Group A: animals constituted the normal
untreated controls
• Group B: Comprised of carcinogen control
animals.
• Dimethylhydrazine(DMH) was administered
subcutaneously, at a dose of 20mg/kg-body
weight ,once a week in 0.9%Na Cl solution(pH
7.2) for a total period of 30 weeks .
• Chemoprotective treatment part(1) Two weeks
then DMH was administered
• Chemoprotective treatment part(2) Four weeks
then DMH was administered

41. Normal untreated control
Fig. 29: No signs of abnormality
0.3% methylene blue stained colon . X 200

42. Putative signs of the induction of cancer in colon:
• Increase in No. of ACF near epithelial surface
• Increase in size of ACF
• ACF are lined with thick, deeply stained epithelial cells
• Nuclei are started to show mitotic changes
• Increase in pericryptal zones

43. DMH Carcinogen Control
Fig. 30: Adenocarcenoma induced by DMH
•Invasion of malignant cells to muscle layers
•Tumor consists of crowded irregular malignat acini separated by thin
fibrovascular stroma
0.3% methylene blue stained colon
a X 200 b X 400

44. Fig. 31: Adenocarcenoma induced by DMH in colon (adenomatous polyps )
0.3% methylene blue stained colon X 200

45. Curcumin was given orally daily for 2
weeks ,then DMH
Fig. 32: Crowding cells at the surface with enlarged and hyperchromatic
nuclei with scattered aberrant crypt foci (ACF) were seen.
0.3% methylene blue stained colon
a X 200 b X 400

46. Ethyl Curcumin was given orally daily
for 2 weeks ,then DMH
Fig. 33: Moderate protection was seen. Crowding cells at the surface
with enlarged and hyperchromatic nuclei with scattered aberrant crypt
foci (ACF) were seen.
0.3% methylene blue stained colon
a X 200 b X 400

47. Compound 3 was given orally daily for
2 weeks ,then DMH
Fig. 34: Few areas of crowding cells at the surface were seen. Few
numbers of aberrant crypt foci (ACF) were seen.
0.3% methylene blue stained colon
a X 200 b X 400

48. Curcumin was given orally daily for 4
weeks ,then DMH
Fig. 35: Few areas of crowding cells at the surface were seen. Few
numbers of aberrant crypt foci (ACF) were seen.
0.3% methylene blue stained colon
a X 200 b X 400

49. Ethyl Curcumin was given orally
daily for 4 weeks ,then DMH
Fig. 36: Beter protection was seen. Few numbers of aberrant crypt foci
(ACF) were seen with less sign of neoplastic changes.
0.3% methylene blue stained colon
a X 200 b X 400

50. Compound 3 was given orally daily for
4 weeks ,then DMH
 No aberrant crypt foci (ACF) were seen.
No focal cell crowding were seen.
Normal globlet cells.
Only some inflammatory cellular infiltration.
Fig. 37: 0.3% methylene blue stained colon
a X 200 b X 400

51. Aberrant Crypt Foci (ACF) Assay
No of ACF/Rat colon Inhibition%
(mean+S.E)
Group No. of rats/G
N.control (A) 10 ------- -----
DMHcontrol(B) 10 100.5+2.54 0
DMH+ curcumin 10 54.3 +1.26 45.98
DMH + ethyl curcumin 10 51.5 +1.20 48.5
DMH + C3 10 37.4 +0.95 62.6
Chemoprotective efficay of different forms of curcumin and curcumin analogs
administered for 2 weeks orally before DMH-induced Aberrant crypt foci(ACF)
in male wistar rats .
N: Normal control received saline s.c. once weekly for 30 weeks.
DMH : was given at a dose of 20mg/kg s.c once weekly for30 weeks.
DMH+C1 :}Curcumin was given at a dose of 53mg/kg orally once daily orally.
DMH +C2 : Ethyl curcumin was given at a dose of 123mg/kg orally once daily .
DMH +C3 : compound 3 was given at a dose of 161 mg/kg orally once daily
C1,C2, C3 were given 2 weeks before induction of cancer colon by DMH.

52. Group No of rats/G No of ACF/ Inhibit
Rat colon(mean+S.E) %
N.control(A) 10 ----- ----
DMH control(B) 10 100.5+2.54 0
DMH+ curcumin 10 47.7+1.38 52.3
DMH+ ethyl curcumin 10 45.8 + 1.71 54.2
DMH+C3 10 ----- 100
N: Normal control received saline s.c once weekly for 30 weeks .
DMH: was given at a dose of 20mg/kg s.c once weekly for 30 weeks.
DMH+ C1: curcumin was given at a dose of 53mg/kg orally daily .
DMH+C2 : ethyl curcumin was given at a dose of 123mg/kg orally daily.
DMH+ C3 : compound 3 was given at a dose of 161mg/kg orally daily.
C1,C2,C3 were given daily for 4 weeks before induction of cancer colon
by DMH.
Chemoprotective efficay of different forms of curcumin and curcumin
analogs administered for 4 weeks orally before DMH-induced Aberrant
crypt foci(ACF) in male wistar rats .

53. Chemoprotective efficacy of different forms of curcumin and curcumin analogs
administered for 2 weeks orally before DMH-induced colonic cancer in rats .
No of rats /G No.of colon tumors 1
N.control(A) 10 -------
DMH control(B) 10 20
DMH+curcumin 10 8
DMH+ethyl curcumin 10 6
DMH+C3 10 2
N: normal control rats received saline s.c. once weekly for 30 weeks.
DMH: was given at a dose of 20mg/kg s.c. once weekly for 30 weeks .
DMH +C1: curcumin was given at a dose of 53mg/kg orally once daily
for 2 weeks.
DMH+C2: ethyl curcumin was given at a dose of 123mg/kg orally once daily
for 2 weeks .
DMH+C3 : compound 3 was given orally at a dose of 161mg/kg once daily

54. Chemoprotective efficacy of different forms of curcumin and curcumin analogs
administered for 4 weeks orally before DMH –induced colonic cancer in rats
No. of rats /G No.of colon tumors/G
N.control 10 ---
DMH control(B) 10 20
DMH+curcumin 10 4
DMH +ethyl curcumin 10 2
DMH+C3 10 --
N: normal control rats received saline s.c. once weekly for 30 weeks .
DMH : was given at a dose of 20mg/kg s.c. once weekly for 30 weeks.
DMH+C1 : curcumin was given at a dose of 53 mg/kg .
DMH+C2 : ethyl curcumin was given at a dose of 123 mg/kg .
DMH+C3 : compound 3 was given at a dose of 161mg/kg .

55. Chemopreventive Effects in DMH-Induced Colon Cancer in
Albino Rats Model
Histopathological Examination of Chemopreventive efficacy of curcumin and
curcumin analogs administered for 2 weeks orally after DMH-induced colonic
cancer in rats
DMH control(B) 10 20
DMH+C1 (C) 10 8
DMH+C2 (D) 10 6
DMH+C3 (E) 10 2
N: normal control rats received saline s.c. once weekly for 15 weeks.
DMH: was given at a dose of 20mg/kg s.c. once weekly for 15 weeks .
DMH +C1: compound 1 was given at a dose of 53mg/kg orally once daily for 2 weeks.
DMH+C2: compound 2 was given at a dose of 123mg/kg orally once daily for 2 weeks
.DMH+C3 : compound 3 was given orally at a dose of 161mg/kg once daily
No. of rats /G No.of colon tumors/G

56. Chemopreventive efficacy of of curcumin and curcumin analogs
administered for 4 weeks orally after DMH –induced colonic
cancer in rats
No. of rats /G No.of colon tumors/G
N.control 10 ---
DMH control(B) 10 20
DMH+C1(F) 10 4
DMH +C2(G) 10 2
DMH+C3(H) 10 --
N: normal control rats received saline s.c. once weekly for 15 weeks .
DMH : was given at a dose of 20mg/kg s.c. once weekly for 15 weeks.
DMH+C1: compound 1 was given at a dose of 53mg/kg.
DMH+C2 :compound 2 was given at a dose of 123mg/kg .
DMH+C3 : compound 3 was given at a dose of 161mg/kg .
No. of rats /G No.of colon tumors/G
N.control 10 ---
DMH control(B) 10 20
DMH+C1(F) 10 4
DMH +C2(G) 10 2
DMH+C3(H) 10 --
No. of rats /G No.of colon tumors/G
N.control 10 ---
DMH control(B) 10 20
DMH+C1(F) 10 4
DMH +C2(G) 10 2
DMH+C3(H) 10 --

57. Conclusion
• In conclusion, the results of this study
suggest that daily supplementation of
Curcumin and Curcumin Analogs has a
positive beneficial effect against
chemically induced colonic preneoplastic
progression in rats induced by DMH,
which provide an effective dietary
chemoprotective and chemopreventive
approach to disease management .
• Best results were given with compound
(3).

58. Conjugation with gold
nanoparticles
• The result of the previous work inhanced us to use the
Targeted drug delivery system (TDDS), especially gold-based
nanoparticles (AuNPs) to be used as a model system in this
work.
• The 1.4 nm Nanogold® particle is a gold compound: it is not
just adsorbed to proteins, like colloidal gold, but covalently
reacts at specific sites under mild buffer conditions.
• The main objective is that, the specific conjugated drug is
targeted to fit and improve their binding affinity, to the specific
receptor that ultimately will trigger the pharmacological
activity as anticancer.
• The other objective is the dual destructive effect on cancer
cells after releasing of gold nanoparticles from the drug.
• In this work, the highly stable mercapto capped gold
nanoparticles were prepared adopting Scheme 1

59. Synthesis and Conjugation with Gold
Nanoparticles
1- Synthesis of biocompatible gold nanoparticles (GNPs) with
different sizes and shapes
2- Physiochemical characterization of the synthesized GNPs using
TEM, XRD, DLS, …etc. techniques
3- Conjugation of the intended drugs with different types of GNPs
(different sizes & shapes)
4- Physiochemical characterization of the synthesized Drug/GNPs
nanocomposite using TEM, XRD, DLS, …etc. techniques to confirm
the formation of the conjugate.
5- Studying the optimization condition such as effect of Temp.,
pH, loading concentrations…etc. to achieve the most probable
(stable) formula of the Drug/GNPs nanocomposite.
6- Studying the rate of drugs release at variable conditions
7- Pharmaceutical formulation of the final product

60. • The superior stability of these particles under the conditions of
varied pH and electrolyte concentrations will be studied.
• In addition to developing synthetic methodology that is free
from reducing agents and elevated temperature, we will prob
the interactions between the gold core and the stabilizer
molecules using Fourier transformed infrared spectroscopy (FT-
IR) technique (will be done at University of Georgia Technology
Labs).
• Long term stability of newly synthesized anti-cancer compounds
capped GNPs may provide improved shelf life. While the
controlled release and sustained delivery of the anti-cancer
drugs from the gold core is demonstrated with the help of GSH
of varying concentrations.
• In vitro and In vivo anti-cancer activity of released compounds
against different types of cancer cells will be investigated in
detail. Reasons behind enhanced anti-cancer activity will be also
addressed in this work.

61. RO
O
H
N
O
OR
O
H
RO
S N
O
OR
S
nanoparticles Au
R'
Au nanoparticle
RO
S
H
N
R'
O
OR
S
H
Au nanoparticles
P2S5
CS2
Scheme 1
R'
R = CH3, CH5
R' = H, COCH3, CONH2

62. Conjugated compounds
with gold nanoparticles

63. Molecular docking studies of tubulin
inhibitors
• Docking Study was performed using the MOE software.
• Downloading the crystal structure of CDK2 enzyme complexes
with inhibitor was carried out from protein data bank website
(PDB).
• Regularization and optimization for protein and ligand were
performed.
• Determination of the essential amino acids in binding site was
carried out and compared with that present in literature.
• The performance of the docking method was evaluated by re-
docking crystal ligand into the assigned active site of to
determine the root mean square deviation (RMSD) value.
• Interactive docking was carried out for all the conformers of
each compound of the test set to the selected active site.
• Each docked compound was assigned a score according to its
fit in the ligand binding pocket (LBP) and its binding mode.

64. ligand podophyllotoxin E Score = -11 K cal/mol

65. Comp. k1 E Score = -9.8 K cal/mol

66. Comp. k2 E Score = -10.9 K cal/mol

67. Comp. k3 E Score = -11.9 K cal/mol

68. Comp. k4 E Score = -11 K cal/mol

69. Comp. k5 E Score = -10.5 K cal/mol

70. Comp. k6 E Score = -10 K cal/mol

71. Conclusion
Comp. k3 E Score = -11.9 K cal/mol

72. 1- U.S. Patent No. 60/670,844, filed April 13,
2005.
Patents and Published Papers
2- Egyptian Patent No. 1131/2008, filed July 3,
2008.
3- European Patent, PCT/EG 2013/000028.

73. 1. Synthesis of Curcumin Analogues as Potential Antioxidant, Cancer Chemopreventive Agents. Khairia M.
Youssef, Magda A. El-Sherbeny, Faiza S. El-Shafie, Hassan A. Farag, Omar A. Al-Deeb, Sit Albanat A.
Awadalla. Arch. Pharm. Pharm. Med. Chem. 2004, 337, 42-54.
2. Synthesis and Antitumor Activity of Some Curcumin Analogs. Part II. Khairia M. Youssef, Magda A. El-
Sherbeny; Arch. Pharm. Chem. Life Sci., 338, 181-189, 2005
3. Synthesis and in vitro antioxidant activity of some new fused pyridine analogs. Mohamed A. Al-Omar, Khairia
M. Youssef, Magda A. El-Sherbeny, Sit Albanat A. Awadalla, Hussein, I. El-Subbagh; Arch. Pharm. Pharm.
Chem. Life Sci. 2005, 338, 181-189.
4. Curcumin Analogs as Anticancer Agents: 1) Preclinical Safety Evaluation in Mice and Rats. 2)
Chemopreventive Effects in DMH-Induced Colon Cancer in Albino Rats Model. Khairia M. Youssef, Azza M.
Ezzo, Moushira I. El-Sayed, Amal A. Hazzaa, Azza H. EL-Medany, Maha Arafa. The International Symposium
on Recent Progress in Curcumin Reseach, September 11-12, 2006 at Yogyakarta, Indonesia.
5. PAC, a novel curcumin analogue, has anti-breast cancer properties with higher efficiency on ER-negative cells
Ensaf M. Al-Hujaily, Ameera Gaafar Mohamed, Ibtehaj Al-Sharif, Khairia M. Youssef and Pulicat S.
Manogaran, et al. Breast Cancer Res Treat (2011) 128:97–107
6. Synthesis of curcumin and ethylcurcumin bioconjugatesas potential antitumor agents Reem I. Al-Wabli
Omaima M. AboulWafa Khairia M. Youssef Med Chem Res (2012) 21:874–890
7. Molecular Modeling of Novel Curcumin Analogs with Anticipated Anticancer Activity: FUE International
Conference on Pharmaceutical Technologies (ICPT) 2012.Iten M. Fawzy, Khairia M. Youssef, Nasser S. M.
Ismail, Khaled A. M. Abouzid.
8. Design and Synthesis and Biological evaluation of Novel Curcumin Analogs with anticipated anticancer
activity. Iten M. Fawzy, Khairia M. Youssef, Nasser S. M. Ismail, J. Gullbo and Khaled A. M. Abouzid. Journal
of Future University in Egypt 2014. In Press.
9. Molecular docking and in silico ADME study of Novel N9-substituted Purines targeting CK1 and abl-tyrosine
kinase. Iten M. Fawzy, Khairia M. Youssef, Nasser S. M. Ismail , Deena S. Lasheen and Khaled A. M.
Abouzid2. FIP conference 2014 at Bangkok.
10. Newly Designed and Synthesized Curcumin Analogs with in vitro Cytotoxicity and Tubulin
Polymerization Activity. Iten M. Fawzy, Khairia M. Youssef, Nasser S. M. Ismail, Joachim Gullbo and
Khaled A. M. Abouzid, Chemical Biology & Drug Design. Early View, Article first published online: 25
NOV 2014.
.

74. Acknowledgement
• Taher Salah, Professor of Nanotechnology, Manager of
Nanotechnology & Advanced Materials Central Lab.
(NAMCL).Regional Center for Food & Feed (RCFF),
Agricultural Research Center (ARC).
• Nasser Saad, Asst. Prof. Department of Pharmaceutical
Chemistry, Future University in Egypt.
• Azza Ahmed, Asst. Prof. Department of Pharmaceutical
Technology, Future University in Egypt.
• Azza M. Ezzo, Pharmacology Dept., Faculty of Medicine, Al-
Azhar University, Cairo, Egypt.
• Maha Arafa, Histopathology Dept., College of Medicine, King
Saud University, Riyadh, Saudi Arabia.
• King Saud University, Saudi Arabia.
• Future University In Cairo.