Zinc oxide nanocomposites with antitumor activity

doi:10.4236/ns.2010.212163

Paper Menu >>

Journal Menu >>

Vol.2, No.12, 1341-1348 (2010)

doi:10.4236/ns.2010.212163

Natural Science

Zinc oxide nanocomposites with antitumor activity

Emma Arakelova1*, Ashot Khachatryan1, Karapet Avjyan1, Zoya Farmazyan1,

Alvard Mirzoyan1, Lilia Savchenko1, Sedrak Ghazaryan2, Flora Arsenyan2

1Scientific Research Division, State Engineering University of Armenia,Yerevan, Armenia;

*Corresponding Author: emma_arakelova@yahoo.com

2Laboratory of Syntheses Derivatives amino acids and peptides, Laboratory of toxicology and chemotherapy IFOC (Institute of Fine

Organic Chemistry), Yerevan, Armenia

Received 25 September 2010; revised 26 October 2010; accepted 30 October 2010.

ABSTRACT

Zinc oxide nanocomposites in the form of

coatings and composite films with antitumor

activity were obtained by deposition of ZnO

nanofilms on surfaces of ethyl ether Salicyli-

dene DL-tyrosine (S1) and ethyl ether Salicyli-

dene DL-tyrosine Cu (II) chelate (S2) by magne-

tron sputtering of Zn target. Ethyl ether Sali-

cylidene DL-tyrosine, Cu (II) chelate of ethyl

ether salicylidene DL-tyrosine reveal some

anticancer properties. Their zinc oxide nano-

composites were obtained in the form of coat-

ings (S1 + ZnO, S2 + ZnO) and composite films

presenting a mixture of polyvinyl alcohol (PVA)

with S1, S2 (S1 + PVA + ZnO, S2 + PVA + ZnO),

for the purpose of increasing anticancer activity.

Considerable increase in antitumor activity re-

veal ZnO nanocomposites with salicylidene

amino acid chelates (as distinct from their

ethers) in the form of S2 + ZnO (47%) and S2 +

PVA + ZnO (48%) in comparison with S2 (20%).

Structural, spectral properties of the salicylidene

amino acids and their ZnO nanocomposites

were studied.

Keywords: Zinc Oxide; Nanofilm;

Magnetron Sputtering; Nanocompo sites;

Antitumor Activity

1. INTRODUCTION

It is known that biologically active materials formed

by deposition of metal oxide nanostructures on the or-

ganic and polymeric systems surfaces reveal an ability to

depress and kill pathogenic biological flora, and their

introduction to organisms prevents “fixing” of viruses

and bacteria to cell walls, improves antibacterial and

antiviral immunity [1-3].

Here, very important is both the choice of inorganic

material for modification of the biologically active ma-

terials surface and the deposition procedure.

Thin films or nanoscale coatings of ZnO nanoparticles

on suitable substrates have excellent electrical, optical,

and medical properties with broad range of applications

for functional coatings, sensors, in optical devices,

transparent electrodes, solar cells and antibacterial, anti-

tumor activity [4-10].

Various methods like chemical, thermal, spray pyroly-

sis, pulsed laser deposition etc. have been developed to

coat ZnO nanoparticles in the form of thin films on solid

supports such as metal, metal oxides, glass or thermally

stable substrates [6,8,11-14].

Among the known methods of obtaining nanocompo-

sitions, magnetron deposition of metal oxide nanofilms

on the organic and polymeric systems surfaces at rela-

tively low temperatures holds a unique position. Stand-out

distinguishing features of the method are ample oppor-

tunities for controlling the nanocomposition formation

process, low deposition temperature (20ºС  T ≤ 100ºС)

and simplified technique of layer-by-layer deposition of

coatings, allowing growth of nanocomposites with pre-

determined structural parameters.

Development of new nanotechnologies is of great

importance for anticancer chemotherapy as the modern

methods of anticancer drug administration in therapeutic

(and slightly hither) doses are accompanied, as a rule, by

development of serious side effects that frequently are

the reasons of termination of the further course of treat-

ment.

It is known that amino acid Schiff’s bases form stable

neutral lipophilic complexes with divalent ions of

d-family metals. Some of them, for example, copper

complexes reveal radioprotective, cytostatic action, in-

hibiting reversibly DNA synthesis [15-19]. It is also

known that in organisms, basic amino acids are binding

agents for metal oxides, in particular for ZnO. Adjusting

and immune effect of ZnO on DNA synthesis, produc-

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

1342

tion of interleukin (IL-2, IL-6, IL-10) and normalization

of cytokine concentration was studied at the patients

with chronic liver disease, in particular, hepatic cirrhosis

[20].

On this basis, formation of thin films ZnO on the sur-

faces of biologically active compounds (aimed at the

surface modification) by magnetron deposition of Zn

target is of special interest.

This work is devoted to deposition of zinc oxide films

on biologically active coatings and composite films of

ethyl ether salicylidene DL-tyrosine, ethyl ether Sali-

cylidene DL-tyrosine Cu (II) chelate with PVA aimed at

modification of their surfaces and obtaining coatings and

composite films with antitumor activity.

The paper discusses the following problems:

1) Obtaining of zinc oxide nanocomposites by deposi-

tion of zinc oxide nanofilms on coatings, composite

films of ethyl ether salicylidene DL-tyrosine and ethyl

ether salicylidene DL-tyrosine Cu (II) chelate by DC-

magnetron deposition of zinc targets.

2) Study of anticancer activity of ethyl ether Sali-

cylidene DL-tyrosine, ethyl ether salicylidene DL-tyrosine

Cu (II) chelate and zinc oxide nanocomposites in the

form of coatings and composite films

3) Structural, spectral properties of salicylidene amino

acid and their chelate compounds, as well as ZnO nano-

composites coatings of S1 + ZnO, S2 + ZnO and PVA

composite films (S1 + PVA + ZnO, S2 + PVA + ZnO).

2. EXPERIMENTAL DETAILS/

METHODOLOGY (SELECT THE MORE

APPROPRIATE)

The paper is aimed at obtaining of zinc oxide nano-

composite coatings and polymer composite films with

anticancer activity by deposition of ZnO nanofilms on

surfaces of ethyl ether salicylidene DL-tyrosine, S1 and

their Cu (II) chelate, S2 by magnetron sputtering of Zn

target. Ethyl ether salicylidene DL-tyrosine and ethyl

ether salicylidene DL-tyrosine Cu (II) chelate are new

compounds for the first time synthesized in the Institute

of Fine Organic Chemistry of National Academy of Sci-

ences of the Republic of Armenia. They were used as

model compounds for obtaining zinc oxide nanocompo-

sitions.

2.1. Formation of Coatings and Composite

Films with PVA from Ethyl Ether

Salicylidene DL-Tyrosine and Ethyl

Ether Salicylidene DL-Tyrosine Cu (II)

Chelate

Coatings from ethyl ether salicylidene DL-tyrosine

and ethyl ether salicylidene DL-tyrosine Cu (II) chelate

were obtained on glass substrates as a paste with di-

methyl sulfoxide.

As solvents, olive oil, ethylene glycol (EG), polyeth-

ylene glycol (PEG), dimethyl sulfoxide (DMSO) were

tested. DMSO was chosen as a low-toxic substance with

antiinflammatory, antimicrobial action, high permeabil-

ity through biological membranes. Pastes from S1, S2

were prepared taking into account their daily maximum

permissible doses for the animals.

Several samples of polyvinyl alcohol differed in mo-

lecular mass and functional composition were used to

create needed polymeric matrix and choose the condi-

tions of obtaining the films prospectively appropriate for

their combination with S1,S2 and zinc oxide. The PVA

samples were: PVA-1, MM approximately 20000, mass

fraction of acetate group 1.3%, PVA-2, MM approxi-

mately 21000, mass fraction of acetate group 10.6%,

PVA-4, MM = 50000-60000, mass fraction of acetate

group 5.3%. PVA-4 differs from PVA-1 and PVA-2 in

higher MM, and its mass fraction of acetate group is

intermediate between PVA-1 and PVA-2.

The composition films from S1 and S2 with PVA were

obtained from 0.5 ml of 5% PVA solution with a meas-

ured amount of S1 and S2 corresponding to therapeutic

doses of S1 and S2 for white nondescript mice. The film

diameter was 2.8 cm (surface area was 6.1 cm2). On the

composite films of S1 and S2 with PVA the ZnO nano-

films were deposited by magnetron sputtering of zinc

target.

2.2. DC-Magnetron Deposition of ZnO

Nanofilms on Surfaces of Salicylidene

Amino Acids and Their Chelates

To deposit nanosize ZnO films on surfaces of coatings

and composite films of ethyl ether salicylidene DL-tyrosine,

ethyl ether salicylidene DL-tyrosine Cu (II) chelate, a

modified UVN-71P3 device, DC-magnetron intended for

sputtering of metal targets was used.

UVN-71P3 device was equipped with a system of

working gas flow measurement and control consisting of

supply and indication unit PR4000F as well as two gas

flow regulators MFC 1179 for Аr and О2 gases.

The substrates from optic glass for deposition of

coatings or composite films from the investigated com-

pounds were fixed inside the device using special clamps

ensuring their movement within the vacuum chamber.

ZnO nanofilm deposition process was carried out by

magnetron sputtering of Zn target at parallel arrange-

ment of the target and substrate. The deposition device is

powered from an energy supply source. To carry out

experiments on ZnO nanofilm deposition within the in-

vestigated temperature interval, a system based on

Peltier element or resistance heater ensuring decrease

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

134

1343

and stabilization of the substrate temperature was used.

ZnO nanofilms were deposited on rotating substrates (to

ensure uniform thickness) at the substrate temperature

range from 20ºС to 100ºС.

When selecting technological regimes of ZnO nano-

film deposition on the substrate (the working gases О2

and Аr ratio, operating current of the magnetron source,

target-to-substrate distance, substrate temperature), the

conditions were considered to eliminate undesirable ex-

traneous transformations of salicylidene amino acids and

their chelates.

To form zinc oxide nanocompositions of the investi-

gated compounds S1, S2 in the form of coatings and

composite films, a technique of ZnO nanofilm deposi-

tion on glass substrates was preliminary developed.

ZnO nanofilms were deposited on glass substrates by

magnetron sputtering of Zn targets (55 mm in diameter)

at the operating current from 80 mA to 500 mA, pressure

of 10-2 mm Hg, and target-to-substrate distance from 7

cm to 13 cm. As working gases, Аr and О2 were used in

the ratio of 70%-30%. To keep clean the deposition

process, the vacuum chamber was exhausted to 2 × 10-6

mm Hg before the film deposition.

2.3. Technique of Identification of Antitumor

Activity of Ethyl Ether Salicylidene

DL-Tyrosine, Their Cu (II) Chelate and

Zinc Oxide Nanocomposites on Strains

of Inoculate Tumors

The study was carried out in toxicology and chemo-

therapy laboratory of Scientific Technological Centre of

Organic and Pharmaceutical Chemistry of the National

Academy of Sciences of the Republic of Armenia under

conditions close to GLP principles and European stan-

dards.

Antitumor coatings and composite films of zinc oxide

nanocomposites of ethyl ether salicylidene DL-tyrosine,

ethyl ether salicylidene DL-tyrosine Cu (II) chelate and

initial compounds were tested on the strains of sarcoma

180 tumors inoculated to white nondescript mice. Pre-

liminary, maximum permissible doses (MPD) were de-

termined by vivisections, and on their basis-therapeutic

doses of the substances, which made as a rule 20% of the

single MPD.

For subinoculation, as a transplant the peaces of tumor

tissues not subject to necrosis and crushed to homoge-

neous mass were used. Physiological salt solution was

added to the tumor in 1:3–1:4 ratio, and the obtained

suspension was introduced to the animals (with initial

weight 22-26 g) subcutaneously by a syringe at a rate of

0.3 ml.

Nanocomposites in the form of ZnO + (S1, S2) coat-

ings were introduced to the animals subcutaneously as a

suspension in DMSO 48 hours after the tumor subin-

oculation on a daily basis during 6 days.

Zinc oxide nanocomposites and initial compounds in

the form of composite films were deposited operation-

ally in sterile conditions (box) by subcutaneous applica-

tions for 6 days through a 5-7 mm in section in spinal

and scapular parts of the animals.

48 hours after the experiment the animals were killed,

the tumors were withdrawn and weighed. Antitumor

activity was determined by the percentage of the tumor

inhibition with respect to the control animals.

2.4. Сharacterization of Ethyl Ether

Salicylidene DL-Tyrosine, Their Cu (II)

Chelates and Zinc Oxide

Nanocomposites

X-ray diffraction (XRD) patterns were obtained on

CAD4, Enraf-Nonius and DRON-2.0 X-ray diffracto-

meters. NMR spectra were recorded on Varian Mercury

300VX spectrometer with all-round frequency 300 MHz.

IR spectra were recorded on NEXUS FT-IR spec-

trometer. Mass spectra were recorded on МХ-1321А.

3. RESULTS & DISCUSSION

3.1. Selection of Optimum Surface Areas for

S1, S2

The ZnO nanofilm surface coatings are S1, S2 deposi-

tions over a certain surface area of object glass. Thick-

ness and optimum ZnO nanofilm surface area was cal-

culated for the case, when its content in the film was

5-10% of that of S1, S2. It is known from publications

that at addition of 5% ZnO to plasma of rabbit with in-

culcated Staphylococcus, antibacterial activity increases

in comparison with that for control samples without ZnO

[20]. On this basis, ZnO content in the film (5-10% of

S1, S2 weight) was taken for rough estimate, with a view

to obtain reliable optimum results. To ensure techno-

logical effectiveness of the deposition process, the sur-

face area was calculated taking into account the amount

of S1 and S2 for daily dose of 6 mice. Suspension of S1 +

ZnO nanocomposite did not reveal reliable antitumor

action (10%) at subcutaneous introduction of m1 = 1.4

mg/mouse of ethyl ether salicylidene DL-tyrosine to

mice, when ZnO deposition area on S1 was s1 = 2.54

cm2. However at the same daily dose, when ZnO deposi-

tion area on S1 coatings were s1 = 9.62 cm2 and s2 = 15.9

cm2, antitumor activity was 30%-32% for d = 100 nm

thick ZnO nanofilms (Table 1).

At subcutaneous introduction of S2 + ZnO, when the

content of ethyl ether salicylidene DL-tyrosine Cu (II)

chelate was m = 1.2 mg/mouse and s = 2.54 cm2, the

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

1344

antitumor effect was only 27%. Taking into account the

calculation for the S1 with optimum result, the ZnO

nanofilm deposition area was calculated for S2 at the

following daily doses: m1 = 1.2 mg/mouse, m2 = 1.4

mg/mouse; m3 = 2.8 mg/mouse. The nanocomposites

were formed on the following surface areas of ethyl

ether salicylidene DL-tyrosine Cu (II) chelate:

s1 = 9.62cm2, s2 = 15.9 cm2, s2* = 19.8 cm2, s3 = 31.8

cm2. Chemotherapeutic investigations have revealed

increase in antitumor activity from 27% to 42% at daily

dose m1 = 1.2 mg/mouse. However with further increase

of ZnO nanofilm surface area, a slight decrease of activ-

ity takes place. At the increase of daily dose to m = 2.8

mg/mouse and selection of optimum area, s = 31.8 cm2,

increase in antitumor activity to 47% was revealed (Ta-

ble 1).

Optimum ZnO nanofilm area coated by ethyl ether

salicylidene DL-tyrosine and ethyl ether salicylidene

DL-tyrosine Cu (II) chelate was determined. Significant

increase in antitumor activity of ZnO nanocomposites of

ethyl ether salicylidene DL-tyrosine Cu (II) chelate in

comparison with the initial compound is observed. It

was revealed that ZnO quantitative content in nanofilms

makes 10% of therapeutic dose of the initial compounds.

3.2. Variation of Structural Characteristics

of PVA in Composite Films with S1, S2

It was revealed at formation of composite films PVA

with S1 and S2 that the films obtained from the mixture

of PVA-1 and S-1 did not dissolve in water after drying;

the films obtained from the mixture of PVA-2 and S-1

(as well as PVA-2 and S-2) dissolve in water; PVA-1

film did not dissolve in water after deposition of ZnO

nanofilms (by magnetron sputtering of Zn target) on its

surface, whereas PVA-2 film with ZnO did dissolve.

Solubility of the films before and after deposition of

ZnO nanofilms was studied both in model conditions at

various pH and in vivo by subcutaneous introduction to

mice and measurement of solubility time.

The following results were obtained: (PVA-1 + ZnO)

film did dissolve in animal organisms, but slowly, during

4 days. However (PVA-1 + S1 + ZnO) film resolved

under the animal skin in 6-7 days (Table 2), i.e. its pro-

longed dissolution in animal organisms was ensured.

(PVA-1 + S2 + ZnO) film did dissolve under the mice

skin.

(PVA-2 + S1) and (PVA-2 + S1 + ZnO) films did dis-

solve 4-5 days, no antitumor effect was revealed.

(PVA-2 + S2) and (PVA-2 + S2 + ZnO) films did dis-

solve in one day, however (PVA-4 + S2) and (PVA-4 +

S2 + ZnO) resolved under the animal skin in 5-6 days

(Table 2).

So, it was revealed that for ethyl ether salicylidene

Table 1. Effect of ZnO nanofilm area surface on antitumor

activity of ZnO nanocomposites.

Composition Daily dose,

mg\mouse

Coated area (S1,S2) of

glass substrates, cm2

(daily dose for 6 mice)

Antitumor

action, %

S1 1.4 - 31

S1 + ZnO 1.4 2.54 10

S1 + ZnO 1.4 9.62 30

S1 + ZnO 1.4 15.90 33

S2 1.4 - 18

S2 2.8 - 20

S2 + ZnO 1.2 2.54 27

S2 + ZnO 1.2 9.62 42

S2 + ZnO 1.4 15.9 42

S2 + ZnO 1.4 19.8 40

S2 + ZnO 2.8 31.8 47

Table 2. Effect of polymer grade on in vivo solubility and an-

titumor activity of ZnO nanocomposite films.

Composition Daily dose,

mg\mouse

In vivo solubility of

films under the mice

skin

Antitumor

action, %

S1 1.4 - 31

S1 + PVA-1 + ZnO1.4 Dissolved, 6-7 days 31

S1 + PVA-2 + ZnO1.4 Dissolved, 4-5 days No effect

S1 + PVA-4 + ZnO1.4 Dissolved, 5-6 days No effect

S2 1.4 - 18

S2 2.8 - 20

S2 + PVA-1 + ZnO1.4 Not dissolved -

S2 + PVA-2 + ZnO1.4 Dissolved, 4-5 days 24

S2 + PVA-4 + ZnO1.4 Dissolved, 5-6 days 42

S2 + PVA-4 + ZnO2.8 Dissolved, 5-6 days 48

DL-tyrosine the most optimum option of the composite

film formation is the use of PVA-1 with molecular

weight 20000, mass fraction of acetate group 1.3%. To

form the composite film with ethyl ether salicylidene

DL-tyrosine Cu (II) chelate, as an intermediate option

PVA-4 was selected with molecular weight 60000,

where the amount of acetate groups (5.7%) is intermedi-

ate between PVA-1 (1.3%) and PVA-2 (10,5%) and mo-

lecular weight is higher.

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

134

1345

3.3. Optimum Technological Parameters of

ZnO Nanofilm Deposition on Coatings

and S1, S2 Composite Films

Optimum technological parameters of zinc oxide nan-

ofilm magnetron deposition on the surface of ethyl ether

salicylidene DL-tyrosine, ethyl ether salicylidene DL-

tyrosine Cu (II) chelate in the form of coatings (S1, S2)

and composite films (PVA + S1, PVA + S2) were deter-

mined.

The substrate material temperature was measured de-

pending on the changes of the magnetron source operat-

ing current at various target-to-substrate distances (Fig-

ure 1(а)). It follows from this dependence that within

the investigated range of operating currents from 80 mA

to 500 mA, the substrate surface temperature was below

100˚С. At such temperatures, there were no undesirable

extraneous transformations of the surface of biologically

active substances S1 and S2. X-ray investigations were

carried out and experimental results on antitumor active-

ity were obtained for ZnO compositions formed at vari-

ous technological regimes of ZnO deposition on the re-

searched biologically active compounds. It was revealed

that the structure of 100 nm thick ZnO nanofilms with

optimum size of nanoparticles (15-30 nm) is appropriate

for interaction with the researched biologically active

compounds in form of both coatings and composite films.

Such nanosize ZnO films were obtained at the following

technological regimes: operating current from 400 mA,

operating voltage 240 V and operating pressure 10-2 mm

Hg. For these regimes, the ZnO nanofilm deposition rate

was studied depending on target-to-substrate distance

(Figure 1(b)). Optimum target-to-substrate distance, 10

cm was selected taking into account the obtained de-

pendences and uniformity of the ZnO films deposited on

the S1, S2 sample surfaces. At such target-to-substrate

distance the film growth rate made approximately 6

nm/min.

Composites on the basis of ZnO nanofilms and bio-

logically active substances S1 and S2 were obtained by

deposition of approximately 100 nm thick zinc oxide

nanofilm (using Zn target) on S1 and S2 deposited in the

form of coatings or composite films, in the following

technological regime: operating current from 400 mA,

gas mixture Ar and O2 at 70:30 ratio; operating pressure

10-2 mm Hg, target-to-substrate distance 10 cm.

3.4. Antitumor Activity of S1, S2 and Zinc

Oxide Nanocomposites

It was revealed as a result of study of acute toxicity

that MPD values for S1 and S2 are 50 mg/kg and 250

mg/kg, correspondingly. In view of these data, as pre-

liminary therapeutic dose for S1 and S2 were used 15

(a)

(b)

Figure 1. (a) Dependence of the substrate material temperature

upon the magnetron source current at various target-to-substrate

distances: 1) 7 cm; 2) 10 cm; (b) Dependence of ZnO nanofilm

deposition rate upon the target-to-substrate distances at various

magnetron source currents: 1) 200 mA; 2) 400 mA.

mg/kg and 50 mg/kg, correspondingly.

In chemotherapeutic experiments S1 at 0.3 mg/mouse

(15 mg/kg) dose did not reveal antitumor effect and

general toxic action, so in subsequent experiment the

dose was increased to 1.5 mg/mouse. It was revealed

that at such dose S1 inhibited sarcoma 180 growth by

31% and did not reveal toxic action on the animal or-

ganism. S2 at therapeutic dose of 1 mg/mouse did not

cause significant inhibition of sarcoma 180 (18%), and

with the dose increase to 2.8 mg/mouse no increase in

therapeutic action was determined (20%).

S1 + ZnO nanocomposite in the form of coatings and

composite films S1 + PVA-1 + ZnO revealed the same

activity as S1: 33% and 31%. However S2 + ZnO nano-

composites where S2 content was 1 or 2.8 mg/mouse,

revealed at subcutaneous injection to animals and film

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

1346

application much higher antitumor activity (in form of

both S2 + ZnO coating, 47%, and S2 + PVA-4 + ZnO

composite films, 48%) than S2(20%).

So, as distinct from nanocomposites with S1 + ZnO,

S2 + ZnO in form of both coatings and composite films

revealed relatively higher therapeutic effectiveness than

at using S2 only.

3.5. Structural, Spectral Characteristics of

S1, S2 and Zinc Oxide Nanocomposites

Complete X-ray diffraction study of ethyl ether Sali-

cylidene DL-tyrosine was carried out using CAD4, En-

raf-Nonius diffractometer. The compound was synthe-

sized for the first time in the Institute of Fine Organic

Chemistry of National Academy of Sciences of the Re-

public of Armenia.

Figure 2 present the molecular structure (Figure 2(a))

and crystal packing on plane ac for three molecules (for

cleanness) (Figure 2(b)) of ethyl ether salicylidene

DL-tyrosine. In the compound’s crystalline structure,

bond lengths (N (2)-C (3): 1.296Å, C (1)-N (2): 1.448 Å)

are typical for C = N Schiff base bond and C-N single

bond, respectively [21]. The compound structure con-

tains intramolecular hydrogen bonds between N (2) and

O (10) atoms, N (2)-H (2)…O (10), 2.565 Å in length

and intermolecular hydrogen bonds O (18)-H (18)…O

(10), 2.623 Å in length.

IR study was carried out on ethyl ether salicylidene

DL-tyrosine Cu (II) chelate and has shown that the

compound includes free hydroxyl group of a salicylidene

fragment in the valence vibration region 3309 cm-1.

In the metal complex, shift of COO- absorption band

was occurred to 1668 cm-1 as distinct from S1 with 1733

cm-1, and a shift –CH = N to 1651 cm-1 due to interact-

tion between Cu and nitrogen on account of free electron

pair.

Also, there were differences in the region of deforma-

tion vibrations: C-O, C-O-H and C-O-C, Me-O, Me-N.

Also, IR study was carried out on ethyl ether Sali-

cylidene DL-tyrosine zinc oxide nanocomposites. The

study has revealed identity of ethyl ether salicylidene

DL-tyrosine zinc oxide nanocomposite and initial S1

spectra. They did not contain expected shifts of absorp-

tion bands of C = O, C-O-H, C-O-C functional groups in

1206-1065 cm-1 region (deformation vibrations) and in

the valence vibration region of OH group (3000-3500

cm-1). ZnO presumably presents in the form of mechanical

mixture with ethyl ether salicylidene DL-tyrosine.

Comparison of the absorption spectra of metal com-

plex and ethyl ether salicylidene DL-tyrosine Cu (II)

chelate zinc oxide nanocomposite has revealed a shift of

absorption band in the valence vibration region of OH

group from 3309 cm-1 in the initial metal complex to

(a)

(b)

Figure 2. Molecular structure (a) and crystal packing on the

plane ac (b) of ethyl ether salicylidene DL-tyrosine.

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

134

1347

3335 cm-1 in metal oxide composite. It is likely that this

shift is related to interaction between ZnO and OH group

of the salicylidene fragment.

In case of ZnO nanocomposite films (S1 + PVA-1 +

ZnO) there is interaction of PVA-1 functional groups

with ZnO and absence of ZnO-S1 interaction. In case of

(S2 + PVA-4 + ZnO) films interactions of ZnO with

functional groups of both PVA-4 and S2 were revealed.

Study of nanocomposite films has revealed the de-

pendence of the interactions with formation of bonds in

the composite film components upon the structure of

S1and S2 compounds.

X-ray investigations of ethyl ether salicylidene DL-

tyrosine have detected presence of intramolecular hy-

drogen bond N-H…O and intermolecular hydrogen

bonds of OH hydroxyl group and carbonyl group, which

form chains in compounds. IR spectral investigations of

ZnO nanocomposite coatings (S1 + ZnO) did not reveal

ZnO-S1 bonds. Spectral investigations of ZnO nano-

composite films (S1 + PVA + ZnO) have revealed inter-

action of PVA functional groups with ZnO and absence

of ZnO-S1 interaction. IR spectral investigations of ethyl

ether salicylidene DL-tyrosine Cu (II) chelate did not

reveal intramolecular or intermolecular hydrogen bonds

in the structure; however ZnO nanocomposite coatings,

S2 + ZnO, reveal obvious formation of intermolecular

hydrogen bond of ZnO...НO type. ZnO nanocomposite

films from PVA with S2 reveal interaction of ZnO with

functional groups of both PVA and S2.

Thus, the surface modification of ethyl ether Sali-

cylidene DL-tyrosine Cu (II) chelate coatings as well of

composite films from these compounds by deposition of

ZnO nanofilms using magnetron sputtering results in

increase of antitumoral activity of the obtained nano-

composites, apparently due to formation of intermolecu-

lar hydrogen bond between ZnO and hydroxyl groups of

these compounds.

4. CONCLUSIONS

1) Zinc oxide nanocomposites of ethyl ether salicyli-

dene DL-tyrosine and ethyl ether salicylidene DL-tyrosine

Cu (II) chelate were obtained by deposition of approxi-

mately 100 nm thick zinc oxide nanofilms on coatings

and composite S1, S2 films using magnetron sputtering

of Zn targets in the temperature range of 20˚С  T ≤

100˚С at operating pressure of 10-2 mm Hg.

2) Optimum ZnO nanofilm area coated by ethyl ether

salicylidene DL-tyrosine and ethyl ether salicylidene

DL-tyrosine Cu (II) chelate was determined. Significant

increase in antitumor activity of ZnO nanocomposites of

ethyl ether salicylidene DL-tyrosine Cu (II) chelate in

comparison with the initial compound is observed. It

was revealed that ZnO quantitative content in nanofilms

makes 10% of therapeutic dose of the initial compounds.

3) Zinc oxide composite films of ethyl ether salicyli-

dene DL-tyrosine and ethyl ether salicylidene DL-tyrosine

Cu (II) chelate with polyvinyl alcohol are obtained en-

suring their prolongation in animals (mice) organizm. It

is shown that for ethyl ether salicylidene DL-tyrosine the

most optimum option of the composite film formation is

the use of PVA with molecular weight 20000 and mass

fraction of acetate group 1.3% To form the composite

film with ethyl ether salicylidene DL-tyrosine Cu (II)

chelate, the optimum option is PVA with molecular

weight 60000 and amount of acetate groups of 5.7%.

4) Chemotherapeutic investigations have shown that

antitumor activity of zinc oxide nanocomposites of ethyl

ether salicylidene DL-tyrosine in the form of both S1 +

ZnO coatings (33%) and S1 + ПВС + ZnO composite

films (31%) was approximately the same as that of

S1(31%).

5) ZnO nanocomposites with Cu (II) chelate of ethyl

ether salicylidene DL-tyrosine (as distinct from their

ester) in the form of S2 + ZnO (47%) coatings and S2 +

PVA + ZnO (48%) composite films have revealed con-

siderable increase in antitumoral activity in comparison

with S2 (20%).

6) X-ray investigations of ethyl ether salicylidene

DL-tyrosine have detected presence of intramolecular

hydrogen bonds and intermolecular hydrogen bonds,

which form chains in the compounds. IR spectral invest-

tigations of ZnO nanocomposite coatings (S1 + ZnO)

did not reveal ZnO-S1 bonds. Spectral investigations of

ZnO nanocomposite films (S1 + PVA + ZnO) have re-

vealed interaction of PVA functional groups with ZnO

and absence of ZnO-S1 interaction.

7) IR spectral investigations of ethyl ether salicylidene

DL-tyrosine Cu (II) chelate did not reveal intramolecular

or intermolecular hydrogen bonds in the structure, how-

ever ZnO nanocomposite coatings, S2 + ZnO, reveal

(judging from the deviation in valence vibrations of OH

group) obvious formation of intermolecular hydrogen

bond of ZnO-H…O type. ZnO nanocomposite films

from PVA with S2 reveal interaction of ZnO with func-

tional groups of both PVA and S2.

8) Surface modification of ethyl ether salicylidene

DL-tyrosine Cu (II) chelate coatings as well as compos-

ite films based on these compounds, by deposition of

ZnO nanofilms using magnetron sputtering resilts in

increase of antitumor activity of the obtained nanocom-

posites, apparently due to formation of intermolecular

hydrogen bonds between ZnO and hydroxyl groups of

these compounds.

5. ACKNOWLEDGEMENTS

This work was supported by Grant А-1563 awarded by the Interna-

E. Arakelova et al. / Natural Science 2 (2010) 1341-1348

1348

tional Science and Technology Center.

Openly accessible at

REFERENCES

[1] Brown, S. (1992) Engineered iron oxide-adhesion mu-

tants of the Escherichia coli phage lambda receptor.

Proceedings of the National Academy of Sciences, USA,

1992, 89, 8651-8655.

[2] Burrell, R.E. (1997) Anti–microbial coating for medical

devices. Patent Application , 1203.

[3] Chule, K., Chule, A.V., Chen, B.-J. and Ling, Y.-C. (2006)

Preparation and characterization of ZnO nanoparticles

coated paper and its antibacterial activity study. Green

Chemistry, 8, 1034-1041.

[4] Wang, Z.L. (2004) Zinc oxide nanostructures: growth,

properties and applications. Journal of Physics: Con-

densed Matter, 16, 829.

[5] Wang, X.D., Summers, C.J. and Wang, Z.L. (2004)

Large-scale hexagonal-patterned growth of aligned ZnO

nanorods for nano-optoelectronics and nanosensor arrays.

Nano Letters, 4, 423.

[6] Shim, E.S., Kang, H.S., Kang, J.S., Kim, J.H. and Lee, S.

Y. (2002) Effect of the variation of film thickness on the

structural and optical properties of ZnO thin films depos-

ited on sapphire substrate using PLD. Applied Surface

Science, 186, 474-476.

[7] Chen, P., Ma, X. and Yang, D., (2006) Fairly pure ultra-

violet electroluminescence from ZnO-based light-emit-

ting devices. Applied Physics Letters. Applied Physics

Letters, 89, 111-112.

[8] Jin, B.J., Woo, H.S., Im, S., Base, S.H. and Lee, S.Y.

(2001) Applied Surface Sci ence, 169-170, 521-524.

[9] Hill, G.M., Mahan, D.C., Carter, S.D., Cromwell, G.L.,

Ewan, R.C., Harrold, R.L., Lewis, A.J., Miller, P.S.,

Shurson, G.C. and Veum, T.L. (2001) Effect of

pharmacological concentrations of zinc oxide with or

without the inclusion of an antibacterial agent on nursery

pig performance. Journal of Animal Science, 79, 34-41.

[10] Hanley, C., Layne J., Punnoose, A., Reddy K.M., Isaac

C., Andrew C., Kevin F. and Denise W. (2008) Increase

in Concentration of a Zn-Containing Volatile Complex by

UV Irradiation of a Target for ZnO Films Synthesis.

Nanotechnology, 19, 10.

[11] Arakelova, E., Grigoryan F., Parvanyan, V., Asatryan, G.

and Antonio, S. (2006) Increase in concentration of a

Zn-containing volatile complex by UV irradiation of a

target for ZnO films synthesis. Texas. EPD Congress

2006, Texas, 2006, 813-818.

[12] Arakelova, E., Parvanyan, F.G.V., Asatryan, G. and Anto-

nio, S. (2006) Study of decomposition regularities for a

Zn-containing volatile complex used in ZnO films syn-

thesis. Texas EPD Congress 2006, Texas, 2006, 883-888.

[13] Xu, W.Z., Ye, Z.Z., Zeng, Y.J., Zhu, L.P., Zhao, B.H.,

Jiang, L., et al., (2006) ZnO light-emitting diode grown

by plasma-assisted metal organic chemical vapor deposi-

tion. Applied Physics Letters, 88, 112-116.

[14] Banerjee, A.N., Ghosh, C.K., Chattopadhyay, K.K., Mi-

noura, H., Sarkar, A.K., Akiba, A. and Kamiya, A. (2006)

Low-temperature deposition of ZnO thin films on PET

and glass substrates by DC-sputtering technique. Thin

Solid Films, 496, 112-116.

[15] Ghazaryan, S.H., Grigoryan, K.P., Ghevondyan, A.I.,

Minasyan, S.H., Tonoyan, V.C., Bajinyan, S.A., Pogho-

syan A.S., Malakyan, M.H. and Sorensen, J. R. S. (2001)

The synthesis of new derivatives of amino acids and pep-

tides: Study of the radioprotective activity. 14th. Interna-

tional Conference on Radiopharmaceutical Chemistry,

Switzerland, 2001, 10-15.

[16] Minasyan, S.H., Ghazaryan, S.H., Tonoyan, V.C., Bajin-

yan, S.A., Grigoryan, K.P., Sorensen, J.R.S. and Greena-

way F.T. (2006) Synthesis, characterization, and meas-

urement of antioxidant reactivity of Salicylidene-

D,L-Tyrosine Ethyl Ester and Copper (II)(Salicylidene-D,

LTyrosine Ethyl Ester)2 in a linoleic acid peroxidation

reaction system. Synthesis and Reactivity in Inorganic,

Metal-Organic, and Nano-Metal Chemistry, 36, 425-434.

[17] Parashar, R.K., Sharma, R.C. and Mohab, G. (1987) Syn-

thesis and biological activity of some copper tridentate

Schiff Buses. In: Sorenson J.R.J. Ed., Biology of copper

complexes. Humana Press, Clifton NJ, 533-540.

[18] Nath, M., Kamaluddin, L. and Cheema, J. (1993) Copper

(II) complexes of salicylidene amino acid Schiff bases as

models. Peroxidase and catalase. Indian Journal of

Chemistry, 32, 108-113.

[19] Picart, L., Goodwin, W.H., Murphy, T.B. and Johnson

D.K. (1982) Zinc regulates DNA synthesis and IL-2, IL-6,

and IL-10 production of PWM-stimulated PBMC and

normalizes the periphere cytokine concentration in

chronic liver disease. Journal of Cell Biochemistry, 6,

172-173.

[20] Akiyama, H., Yamasaki, O. Kanzaki, H., Tada, J. and

Arata, J. (1998) Effect of zinc oxide on the attachment of

Staphylococcus aureus strains. Journal of Dermatologi-

cal Science, 17, 67-74.

[21] Zhou, G.D. and Duan L.Y. (1995) Basis of structural

chemistry. 2nd Edition, Perking University Publishing

Corporation, Beijing, 249-250.