Drugs R D 2004; 5 (3): 125-139

REVIEW ARTICLE 1174-5886/04/0003-0125/$31.00/0

2004 Adis Data Information BV. All rights reserved.

Fig. 3. Estimation of opacity of the lens by quantitative

morphometric analysis: (a) immature cataract, image of lens with
binary isolation of zone with second degree of optical density;
(b) immature cataract; and (c) almost mature cataract. Optical den-
sities (OD) of different zones of the lens subdivided in diminishing
order into 10 degrees corresponding to different degrees of opacity
of the lens (scale of OD is given at the left margin of the photos).
Numbers on photographs express absolute area of zones in rela-
tive units with assigned value of OD. Dark areas correspond to a
higher value of OD. The test material consisted of opaque human
lenses at different cataract stages obtained during operation by
intracapsular cryoextraction.
126 Babizhayev et al.

Lipid Peroxidation and Cataracts

N-Acetylcarnosine as a Therapeutic Tool to Manage
Age-Related Cataracts in Human and in Canine Eyes
Mark A. Babizhayev,1,2 Anatoly I. Deyev,3 Valentina N. Yermakova,2 Igor V. Brikman2
and Johan Bours4
1 Innovative Vision Products Inc., County of New Castle, Delaware, USA
2 Moscow Helmholtz Research Institute of Eye Diseases, Moscow, Russian Federation
3 Russian State Pirogovs Medical University, Moscow, Russian Federation
4 Institute of Experimental Ophthalmology, University of Bonn, Bonn, Germany

Contents
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
1. Cataracts and Blindness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
2. Complications of Cataract Surgery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
3. Morphological and Molecular Aspects of Lens Transparency . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
4. Lipid Peroxidation and Cataracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
5. Carnosine and N-Acetylcarnosine for Vision . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
6. N-Acetylcarnosine as an Antioxidant for Ophthalmic Application: Mechanism of Action . . . . . . . . 135
7. Clinical Studies of N-Acetylcarnosine for the Treatment of Cataracts . . . . . . . . . . . . . . . . . . . . . . . . . . 136
8. Treatment of Age-Related Cataracts in Canines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
9. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Abstract Cataract formation represents a serious problem in the elderly, with approxi-
mately 25% of the population aged >65 years and about 50% aged >80 years
experiencing a serious loss of vision as a result of this condition. Not only do
cataracts diminish quality of life, they also impose a severe strain on global
healthcare budgets. In the US, 43% of all visits to ophthalmologists by Medicare
patients are associated with cataract. Surgery represents the standard treatment of
this condition, and 1.35 million cataract operations are performed annually in the
US, costing $US3.5 billion (year of costing, 1998).
Unfortunately, the costs of surgical treatment and the fact that the number of
patients exceeds surgical capacities result in many patients being blinded by
cataracts worldwide. This situation is particularly serious in developing countries;
worldwide 17 million people are blind because of cataract formation, and the
problem will grow in parallel with aging of the population. In any event, surgical
removal of cataracts may not represent the optimal solution. Although generally
recognised as being one of the safest operations, there is a significant complica-
tion rate associated with this surgical procedure. Opacification of the posterior
lens capsule occurs in 3050% of patients within 2 years of cataract removal and
requires laser treatment, a further 0.8% experience retinal detachments, approxi-

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts 127

mately 1% are rehospitalised for corneal problems, and about 0.1% develop
endophthalmitis. Although the risks are small, the large number of procedures
performed means that 26 000 individuals develop serious complications as a result
of cataract surgery annually in the US alone. Thus, risk and cost factors drive the
investigation of pharmaceutical approaches to the maintenance of lens trans-
parency.
The role of free radical-induced lipid oxidation in the development of cataracts
has been identified. Initial stages of cataract are characterised by the accumulation
of primary (diene conjugates, cetodienes) lipid peroxidation (LPO) products,
while in later stages there is a prevalence of LPO fluorescent end-products. A
reliable increase in oxiproducts of fatty acyl content of lenticular lipids was shown
by a direct gas chromatography technique producing fatty acid fluorine-substi-
tuted derivatives. The lens opacity degree correlates with the level of the LPO
fluorescent end-product accumulation in its tissue, accompanied by sulfhydryl
group oxidation of lens proteins due to a decrease of reduced glutathione concen-
tration in the lens. The injection of LPO products into the vitreous has been shown
to induce cataract. It is concluded that peroxide damage of the lens fibre mem-
branes may be the initial cause of cataract development.
N-acetylcarnosine (as the ophthalmic drug Can-C), has been found to be
suitable for the nonsurgical prevention and treatment of age-related cataracts. This
molecule protects the crystalline lens from oxidative stress-induced damage, and
in a recent clinical trial it was shown to produce an effective, safe and long-term
improvement in sight. When administered topically to the eye in the form of
Can-C, N-acetylcarnosine functions as a time-release prodrug form of
L-carnosine resistant to hydrolysis with carnosinase. N-acetylcarnosine has poten-
tial as an in vivo universal antioxidant because of its ability to protect against
oxidative stress in the lipid phase of biological cellular membranes and in the
aqueous environment by a gradual intraocular turnover into L-carnosine.
In our study the clinical effects of a topical solution of N-acetylcarnosine
(Can-C) on lens opacities were examined in patients with cataracts and in
canines with age-related cataracts. These data showed that N-acetylcarnosine is
effective in the management of age-related cataract reversal and prevention both
in human and in canine eyes.

1. Cataracts and Blindness
Cataract, the opacification of the eye lens, is the
leading cause of blindness worldwide,[1] accounting
for approximately 42% of all blindness.[1] More than
17 million people are blind because of cataract and
28 000 new cases are reported daily worldwide.[2] In
the US, more than 1.3 million cataract operations are
performed annually at a cost of $US3.5 billion (year
of costing, 1998).[1] Cataract is the leading cause of
functional impairment among the elderly in the US,
and is the most commonly performed surgical pro-
cedure in people aged >65 years in the US.[3] Forty-
three percent of all visits to ophthalmologists by
Medicare patients are associated with cataract.[1]
Approximately 25% of the population aged >65
years and about 50% aged >80 years have serious
loss of vision because of cataract. Since the popula-
tion aged >55 years is most susceptible to lens
opacification and is expected to increase 4-fold
worldwide and significantly in the US,[4] cataract is
a major disease in terms of both numbers of people
involved and economic impact.

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
128
Of the 17 million cases of blindness in the world,
half are in the developing countries of Africa and
Asia. Published data estimate that 1.2% of the entire
population of Africa is blind, with cataract causing
36% of this blindness.[5] In developing countries,
there are simply not enough surgeons to perform
cataract operations. Therefore, a significant number
of people developing cataract become permanently
blind. The disease is becoming more frequent with
increases in the lifespan of this population. While
the majority of cases occur in older age groups,
young individuals are not exempt and, in them, the
rate of maturation is faster. Cataract is far more
prevalent in India and other developing countries
than in Western countries,[6] and it is reasonable to
hypothesise that there could be a nutritional cause of
cataract to explain this difference. Early studies
showed that people who ate food of poorer quality
were indeed more at risk of cataract,[7,8] an index of
poverty consistently identified as a risk factor of
cataract in many different populations.[6]
2. Complications of Cataract Surgery
While cataract surgery is generally recognised as
being one of the safest operations, there is a signif-
icant complication rate. At least 510 million new
visually disabling cataracts occur yearly, with com-
plications of modern surgical techniques resulting in
100 000200 000 irreversibly blind eyes. In the US,
300 000400 000 new visually disabling cataracts
occur annually, with complications of modern surgi-
cal techniques resulting in at least 7000 irreversibly
blind eyes. From 30% to 50% of all patients in the
US having cataract extraction develop opacification
of the posterior lens capsule within 2 years and
require laser treatment, with its own significant risk
of complications.[9] Since the number of cataract
operations is so large, even a small percentage of
complications represents a significant number of
people. Of patients having cataract surgery, 0.8%
developed retinal detachments,[10] 0.61.3% were
rehospitalised for corneal oedema or required cor-
neal transplantation[11] and about 0.1% presented
with endophthalmitis.[10] Thus, aside from secon-
dary cataract, about 2% of 1.3 million people, or
2004 Adis Data Information BV. All rights reserved.
Babizhayev et al.
26 000 individuals, in the US annually develop seri-
ous complications as a result of cataract surgery.
Most morbidity associated with senile cataracts
occurs postoperatively. While the risk of death as a
result of cataract extraction is almost negligible,
studies have shown an increased risk of mortality in
patients who underwent surgery. In a comparison of
167 patients aged 50 years who underwent cataract
extraction at the New England Medical Center over
a period of 1 year with 824 patients who elected one
of six other surgical procedures, it was found that
the former had almost twice the mortality of the
latter. Further analysis showed no significant corre-
lation between diabetes mellitus and increased mor-
tality.[12] Age-related cataract was reported to be
associated with increased risk of death. After adjust-
ment for age, sex and other mortality risk factors,
mixed cataracts with a nuclear/posterior subcapsular
component were significantly associated with
higher risk of death by Cox proportional hazards
regression analyses. These findings are compatible
with the hypothesis that mixed types of cataract with
a nuclear/posterior subcapsular component are in-
dicators of accelerated aging.
The large and growing number of people who are
blind with cataract and the significant complication
rate should be sufficient reason to consider the
search for a medicinal cure of cataracts. The consid-
erable discomfort experienced by patients as their
vision diminishes and the complete loss of accom-
modation resulting from removal of the lens should
also be recognised. Besides possible complications,
an artificial lens just does not have the overall opti-
cal qualities of a normal lens. Identification of the
risk factor(s) and unravelling the mechanism(s)
through which the human eye loses its transparency
and turns opaque would help to develop an effective
preventative approach to cataract blindness.
3. Morphological and Molecular
Aspects of Lens Transparency
The normal human eye lens is a transparent, pale
yellow, resilient, biconvex body enclosed in an elas-
tic capsule. It is suspended behind the pupil of the
eye by strands that fuse with the capsule. The lens is
Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts
a relatively simple tissue that has only a single layer
of epithelial cells on the anterior side of the tis-
sue.[13,14] The lens grows throughout life from the
epithelium that lines the inner surface of its transpar-
ent capsule. The disciform eye lens continues to
grow throughout life. The fresh fibres, developing
by elongation from cubic anterior epithelial cells,
are continually laid down in the equatorial zone. The
older cells in the anterior and posterior cortex and
supranuclear cortex gradually lose their nuclei and
become more and more compressed in the centre or
nucleus of the lens. There is a growing awareness
that the interaction of tissues of the eye influences
the overall viability of the lens organ. Both develop-
ment and cataract disease are affected by the envi-
ronment of the respective tissues of the eye. The
water content is lowest in the nucleus and highest in
the cortex of the lens.[15] The whole lens consists of
about 65% water and 35% proteins. These structural
proteins are called crystallins and form most of the
dry weight of the lens.[6] Transparency of the normal
eye lens depends on the short range order that exists
in the supramolecular organisation, which results in
0 10
B B
0.5
Optical density
I II III
0.25 C
C A
A D
D A
A
Fig. 1. The dynamics of the accumulation of the primary lipid peroxidation molecular products in the lens opacification. = lens opacity
degree (%); IVI = cataract stages, A 190nm, B 206nm, C 230nm (diene conjugates maximum), D 274nm (triene conjugates
maximum), E 330nm.
2004 Adis Data Information BV. All rights reserved.
129
interference effects.[16-18] The conformational
change during cataractogenesis leads to the forma-
tion of aggregates that scatter light, producing
opacification.[6,19] For that reason, an aggregation
process was proposed as the main contributor to
cataractogenesis.[20] Traditionally, the lens is con-
sidered as a sac filled with proteins. Consequent-
ly, the majority of investigators until now have been
searching for the cause of lens high-molecular-
weight protein aggregate formation in the crystalline
physicochemical properties alteration, in the reduc-
tion of protein solubility and in the change of their
amino acid content.[21-23]
Biochemical characteristics of cataract manifes-
tation are as follows: formation of large, high-mo-
lecular weight aggregates of low solubility in the
lens tissue; appearance of blue fluorescence of
non-tryptophan nature; and disintegration of the lens
fibre plasma membranes.[23,24] At the same time, it is
well known that protein aggregate formation in the
lens is accompanied by an intrusion of lenticular
fibre membrane fragments into the cytoplasmic
1055 5564 64100 100
B B B B
IV V VI
C
C
C
A A
D C
D
D
D
E E E E E
Wavelength 330190nm
Drugs R D 2004; 5 (3)
130 Babizhayev et al.

Table I. Content of lipid peroxidation products in human lenses (data presented as mean SD)
Cataract stage (%) n C OD232/OD206 C OD274/OD206 Fl
(AU) (AU)
Transparent lens 0 18 1.9 0.2 0.30 0.02 0.45 0.05 0.07 0.01 18 5
p < 0.1 p < 0.1 p < 0.1 p < 0.1 p > 0.1
Incipient <10 6 2.4 0.2 0.37 0.03 0.58 0.10 0.09 0.01 19 7
Immature 1055 36 2.5 0.1 0.39 0.015 0.67 0.05 0.13 0.01 45 8
p < 0.01 p < 0.01 p < 0.01 p < 0.01 p < 0.02
Almost mature 5564 26 2.9 0.2 0.45 0.03 0.80 0.06 0.16 0.01 53.4 12.0
p < 0.01 p < 0.01 p < 0.01 p < 0.01 p < 0.02
Mature 64100 53 2.4 0.1 0.36 0.025 0.66 0.06 0.11 0.01 74.1 13.2
p < 0.05 p < 0.1 p < 0.02 p < 0.1 p < 0.01
Hypermature ~100 9 2.2 0.2 0.35 0.04 0.45 0.07 0.08 0.01 97.8 10.0
p > 0.1 p > 0.1 p > 0.1 p > 0.1 p < 0.01
= degree of lens opacity; AU = arbitrary units; C = diene conjugates concentration; C = triene conjugates (cetodienes) concentration, C~
OD274/CL; C~ OD232/CL,OD232, OD274, OD206 optical density at 232nm, 274nm and 206nm, respectively; CL = lipid content (mg/mL); Fl =
fluorescence intensity of lipid extract in relative units; n = number of examined lenses; p < 0.05 = reliable values (compared with transparent
lenses).

fraction.[24,25] The above-mentioned aggregates were tion reactions of the 3-hydroxykynurenine or N-

shown to contain phospholipids that can be found in
the lens cytoplasmic protein fraction using phos-
phorus registration.[26] Since in the transparent lens
most lipids localise in the membranes, we have
suggested that processes inducing damage of the
lenticular fibre plasma membrane lipid bilayer par-
ticipate in the aggregation of crystallins.[27]
4. Lipid Peroxidation and Cataracts
Among endogenous processes that can cause in-
jury to membranous structures of cells and tissues,
one of the most important is lipid peroxidation
(LPO). Reactive oxygen species (ROS) is the term
usually used to indicate components that can cause
oxidative stress. It includes oxygen radicals such as
superoxide (O2.) and hydroxyl radical (OH), as
well as oxidants such as hydrogen peroxide (H2O2)
and singlet oxygen (O12 [1g]).[28] During aging
and, in particular, during the development of senile
cataract, activity of enzymatic (superoxide dis-
mutase, glutathione peroxidase and catalase) and
nonenzymatic (ascorbate, cysteine and glutathione)
antioxidant systems in the lens and aqueous humour
are reduced.[29-31]
Accumulation of LPO products in the lens may
be facilitated by the presence of compounds in the
lens that are photosensitisers of free-radical oxida-
formylkynurenine type, which absorb light in the
near-UV region of the spectrum (360400nm).[32]
Two major types of processes can occur with pro-
teins. The first of these involves direct photo-oxida-
tion arising from the absorption of UV radiation by
the protein, or bound chromophore groups, thereby
generating excited states (singlet or triplets) or radi-
cals via photo-ionisation. The second major process
involves indirect oxidation of the protein via the
formation and subsequent reactions of singlet oxy-
gen generated by the transfer of energy to ground
state (triplet) molecular oxygen by either protein-
bound or other chromophores.[32] The photo-oxida-
tion products of tryptophan under the influence of
light can generate active forms of oxygen and prod-
ucts of its successive single-electron reduction (sin-
glet oxygen, superoxide anion radicals, hydrogen
peroxide and hydroxyl radicals), which can be found
in the lens tissue and also in the aqueous humour.[33]
Several varieties of cataract have been described in
the literature; their mechanisms of development
have been linked with the generation of active forms
of oxygen. They include psoralene cataract, cataract
induced by the action of hyperbaric oxygenation,
and cataract arising in animals fed with the catalase
inhibitor 3-amino-1H,1,2,4-triazole.[34-36] Free-radi-
cal enhancers, diquat injected intravitreally as a

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts
III
80
70
60
Fluorescence (relative units)
II
50
40
30
I
20
10
400 440 480 520
Wavelength (nm)
Fig. 2. Characteristic fluorescence spectra of the lipid extracts from
human lenses by excitation wavelength of 365nm; I = norm; II =
almost mature cataract; III = mature cataract. The content of end-
fluorescent lipid peroxidation products was determined from the
lipid extract fluorescence intensity at 365nm excitation and
420440nm emission wavelengths, measured on a HITACHI-
MPF-4 spectrofluorometer (Japan).
single dose (300 nmol in 30L of isotonic saline) in
the right eyes of 5-week-old Dutch belted rabbits
induced early cataract after 2472 hours.[37] The
lenses of the contralateral control eyes injected with
isotonic saline had no change.
On the basis of these facts it has been postulated
that LPO may play a role in the aetiology and
pathogenesis of cataract. Direct proof of LPO acti-
vation has been obtained in cataracts.[27,38-40] The
results of determination of the LPO of different
molecular products revealed in the lipid extracts
from the lens are shown in figure 1 and table I. In
UV absorption spectra of lipid extracts from lenses
with a cataract, there are two additional maxima at
2004 Adis Data Information BV. All rights reserved.
131
230 and 274nm (figure 1). The first of these corre-
sponds to absorption of diene-conjugated structures.
The maximum at 274nm corresponds to triene con-
jugates secondary molecular LPO products. The
results of the determination of diene conjugates in
lipid extracts from the lenses at different stages of
cataract maturation are presented in figure 1. It is
evident that the content of the hydroperoxides hav-
ing conjugated double bonds and determined by a
characteristic maximum in UV spectrum at 230nm
increases at the initial stages of the opacification up
to almost the mature stage of cataract (5564%
opacification degree), assessed by a quantitative
morphometric analysis technique (figure 2). How-
ever, at further stages of mature and hypermature
cataract the lipid peroxidation primary molecular
product level drops a little. At the same time, deter-
mination of the content of fluorescent end-products
of the LPO-Schiff bases determined by lipid extract
fluorescence intensity at 430nm (fluorescence exci-
tation 365nm) revealed an increase monotonously
along with cataract development (figure 3). The
fluorescent end-LPO product concentration in the
lens correlated strongly with the degree of opacity
(r = +0.956, p < 0.01).[40]
An important regularity was revealed in our stud-
ies:[27,38-40] accumulation of the LPO products de-
pends on the stage of cataract development, but not
on its kind (age-related, complicated, diabetic), sug-
gesting a universal role of LPO in the lens opacifica-
tion process. Determination of total thiols and the
content of LPO products for the same lenses showed
that with the development of cataract, the level of
the total thiols of the lens fibres rapidly falls with
intensification of the free-radical oxidation reactions
of lipids.[40] The presence of high correlations be-
tween the degree of clouding of the lens, the level of
the final lipid peroxidation products, and the con-
centration of reduced glutathione and total thiols in
the lens showed that the transparency of the lens, the
concentration of the lipid peroxides and the thiol
level in it were interrelated parameters of a single
process.[40] The content of the polyunsaturated fatty
acids in the lens is rather moderate; hence, direct
registration of their decrease in the course of lipid
Drugs R D 2004; 5 (3)
132 Babizhayev et al.

A
fraction from transparent and cataractous (mature
cataract) human lenses are shown. Although in ma-
I ture cataract no new peaks are found to appear on
gas chromatographic profiles of the lens lipid sam-
ple, the intensity of peaks markedly differs from the
norm as a quantitative ratio. However, a reliable
increase in the intensity of the peaks, whose reten-
Response

tion time does not correspond to the fatty acids

nonoxidised standards (table II), most probably re-
flects the increase in content of oxiderivative poly-
unsaturated fatty acids in cataracts.
The study of cataract formation raises an interest-
A ing question: can modification of the lipids that are
the minor component (only 2% of lens wet mass)
II lead to conformation and solubility changes of the
proteins constituting 35% of the lens wet mass? In
other words, whether the LPO process is a factor
that, together with other metabolic shifts, only ac-
companies cataract development, or, in a number of
cases, whether the LPO activation can be an actual
initiating cause of the lens opacification.[42] In our
Response

study the possibility of cataract induction with phos-

Table II. Analysis of fluorinated derivatives of fatty acids in trans-

parent and cataractous human lensesa
Retention time Concentration (%) p-Value
(min) transparent mature
lens (n = 5) cataract (n = 7)
17.32 2.08 0.10 0.65 0.10 <0.01
35.26 2.17 0.10 10.89 1.10b <0.01
0 45 90 35.76 2.34 0.10 1.55 1.00 >0.1
Retention time (min) 45.61 42.6 1.1 40.7 1.1 >0.1

Fig. 4. Gas chromatograms of the lipid fraction from transparent (I) 59.83 1.27 0.10 2.83 0.10b <0.01
and cataractous (mature cataract) [II] lenses after fluorination. For 63.67 19.56 1.00 0.70 0.10 <0.01
selective determination in the lens lipid fraction of the substances 67.68 1.6 0.1 0.6 0.5 >0.1
containing oxigroups, fluorine-substituted derivatives of the fatty 68.7 2.61 0.10 4.0 0.1b <0.01
acids were obtained.
73.93 2.0 0.1 7.1 0.1b <0.01
74.35 9.1 0.1 0.6 0.1 <0.01
peroxidation is quite difficult. Evidently, to measure 78.07 1.8 0.1 0.6 0.2 <0.01
directly an increase of oxiproducts in unsaturated 82.17 1.8 0.2 0.6 0.2 <0.01
fatty acids is of rather greater importance than to 84.95 1.4 0.1 0.6 0.2 <0.01

reveal a decrease in the fatty acid content. As a result 85.37 5.7 0.1 0.6 0.3 <0.01
86.1 2.1 0.1 0.6 0.2 <0.01
of the fluorination procedure of the lipid oxiproducts
94.58 1.8 0.2 2.2 0.2b >0.1
in human lenses, the in vivo LPO process in human a Oxyproducts accumulation ratio in comparison with
cataracts has been monitored using a gas chromato- transparent lens is 17.2%.
graphy technique with electron-capture detector b Chromatographic peaks characterising accumulation of fatty
acid oxiproducts.
(figure 4).[41] Typical chromatograms of the lipid

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts
a this study by characteristic fluorescence of lipid
b showed decreased activity of glutathione reductase,
Fig. 5. Induction of posterior subcapsular cataract in the rabbit by
injection of the phospholipid liposomes into rabbit vitreous; 0.4mg
injection of (a) oxidised phospholipids, and (b) saturated phospho-
lipids.
pholipid peroxidation products was examined by the
injection of oxidised liposomes into the rabbit vitre-
ous (figure 5).[42] Auto-oxidised phospholipids in-
duced cataract after their injection with the concom-
itant accumulation of the LPO products in the lens
(table III) and the reduction of the lenticular gluta-
thione concentration (table IV). Injection of saturat-
ed phospholipids resistant to peroxidation in the
same concentrations did not induce any lens opacifi-
cation (figure 5b). This is evidence of the initiating
effect of the LPO products in cataract formation.[42]
2004 Adis Data Information BV. All rights reserved.
133
Such cataract modelling is based on a type of cloud-
ing of the crystalline lens similar to that observed in
cataract (posterior subcapsular cataract), resulting
from diffusion of toxic lipid peroxidation products
from the retina to the lens through the vitreous body
on degeneration of the photoreceptors.[42]
Our modelling of cataract was based on polymer-
isation of the secondary LPO products of the di-
aldehyde type presented in the vitreous with the lens
crystallins. Dialdehyde bifunctional reagents can in-
teract with free amino groups forming fluorescent
products of the Schiff-base type and, subsequently,
inter- and intramolecular cross-links. Phospholipid
intramolecular cross-links have been registered in
extracts from cataractous lenses.[27,38-40,42] It is im-
portant that the accumulation of rather small
amounts of oxidised lipids in the lens is sufficient to
induce cataract development. Preventing accumula-
tion of peroxide compounds in the lens and main-
taining a high level of reduced glutathione can prob-
ably prevent further development of cataract and, in
a number of cases, may even partially restore the
transparency of the already opacified lens. The crys-
talline lens can withstand the oxidative stress-in-
duced damaging influences, neutralising the pro-
oxidant agents from the surrounding aqueous hu-
mour in the eye.[43,44] Human cataractous lenses
glutathione peroxidase (catalysing reduction of
organic hydroperoxides including hydroperoxides
of lipids) and superoxide dismutase, but no signs of
depletion in activity of catalase or glutathione per-
oxidase (utilising hydrogen peroxide) [table V].[31,44]
Our findings indicated an impairment in peroxide
metabolism of the mature cataractous lenses com-
pared with normal lenses to result from a deficiency
of reduced glutathione (GSH). An oxidative stress
induced by accumulation of LPO products in the
lens membranes during cataract progression can be
considered to be a primary cause of GSH deficiency
and disturbance of the redox balance in the lens. It
seems essential to improve the capacity of the lens to
withstand oxidative stress induced by lipid perox-
ides, since the opacified lenses have reduced activity
Drugs R D 2004; 5 (3)
134 Babizhayev et al.

Table III. Fluorescent products of lipid peroxidation in eye tissues by cataract modelling with liposomes (fluorescence relative units
excitation/emission: 365/438nm). The data presented (mean SD) were obtained by intraocular injection of 0.4mg of phospholipids; the
material concentration in the samples was equalised by lipid concentration (calculated by characteristic lipid extract absorption at 206nm)
[all units are fluorescence relative units]
Tissue Norm Nonoxidised liposomes: Oxidised liposomes: Saturated liposomes:
(n = 10) 2.0 nmol MDA/mol 22.2 nmol MDA/mol 1 nmol MDA/mol
of phospholipids of phospholipids of phospholipids
(n = 7) (n = 10) (n = 6)
Lens 96.5 23.1 154.4 17.6 173.0 37.4 100.0 11.2
Vitreous 82.2 28.5 326.3 134.0 173.3 37.6 85.1 11.6
Aqueous humour 176.3 25.0 232.0 55.1 240.0 14.0 171.0 7.9
MDA = malonyldialdehyde; n = the number of eyes examined.

of lipid peroxidases that would use glutathione as a
substrate.[43,44]
5. Carnosine and N-Acetylcarnosine
for Vision
Recently it was discovered that some natural
compounds of a peptide character or their metal
chelates may be among the most potent lipoperox-
idase mimetics that have ever been character-
ised.[43-45] L-carnosine and its ophthalmic prodrug
N-acetylcarnosine are part of this group of products.
N-acetylcarnosine, like its parent compound,
carnosine, occurs naturally throughout the human
body. Both compounds are found primarily in the
heart and skeletal muscles (the word carnosine is
derived from the Latin word for flesh) and in the
brain. Research with N-acetylcarnosine demonstra-
ted that it is effective not only in preventing cata-
racts but also in treating them.[46-49] It has been
shown to improve vision by partially reversing the
development of the cataract, thus increasing the
transmissivity of the lens to light. The structural
difference between N-acetylcarnosine and carnosine
is that one hydrogen atom in carnosine replaces an
acetyl group (CH3CO-) and this substitution occurs
at a nitrogen atom. An important chemical differ-
ence between carnosine and N-acetylcarnosine is
that carnosine is relatively insoluble in lipids (fats
and fatty compounds), whereas N-acetylcarnosine is
relatively soluble in lipids (as well as in water). This
means that N-acetylcarnosine may pass through the
lipid membranes of the corneal and lens cells more
easily than carnosine, and may thereby gain access
more readily to the cells interior, which is primarily
aqueous. The advantage of these compounds as uni-
versal antioxidants resides in their ability to give
efficient protection against lipid peroxidation both
in the lipid phase of biological membranes and in the
aqueous environment.[44,50-52] Various protective an-
tioxidant enzymes such as superoxide dismutase or
catalase can react with their substrates only in an
aqueous environment.[50] These molecules are en-
dowed with lipid peroxidase- and hydroxyl radical-

Table IV. Thiol content in the lens by cataract modelling with liposomes. Protein molecular weight was assumed to average 20 000 (data are
presented as mean SD)
Liposomes Phospholipid content n Total thiols Reduced glutathione
(mg) (mol/protein mol) (nmol/mol of initial protein)
Nonoxidised, 0.4 7 2.92 0.34 450 116
unsaturated
1.5 8 2.99 0.37 251 75
Oxidised 0.4 10 2.96 0.24 403 63
1.5 8 3.10 0.47 121 52
Saturated 0.4 6 3.2 0.5 520 71
1.5 3 3.1 0.6 500 102
Controls 10 4.1 0.4 705 106
n = number of eyes examined.

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts 135
Table V. Activity of antioxidative enzymes and rate of decomposition of hydrogen peroxide (H2O2) by transparent lenses and lenses affected by cataract (data are presented as

NADPH = nicotinamide-adenine dinucleotide phosphate; TBHP = tert-buytlhydroperoxide. * p < 0.001; ** p < 0.01; *** p < 0.1; p < 0.05; p < 0.02 (n = 15) compared with
scavenging antioxidant activities as demonstrated in
our studies.[45] For comparison, the classic selenium-
0.404 0.049***

dependent glutathione peroxidase lowers the level

0.195 0.047*
0.51 0.04

of lipid hydroperoxides in cell membranes only

under their stripping (cleavage) with phospholipase
TBHPf

A2.[53] It appears that lipid peroxidase mimetics may

show promise in the therapy and prophylaxis of
cataracts.
0.98 0.21

0.92 0.21
glutathione
peroxidase
1.3 0.2

6.N-Acetylcarnosine as an Antioxidant
for Ophthalmic Application: Mechanism
of Action
glutathione reductasee

Previously published data suggest that L-

carnosine has excellent potential to act as a natural
0.111 0.020

0.059 0.015*
0.355 0.083

antioxidant with hydroxyl radical- and singlet oxy-

gen-scavenging activities.[43,45,54] Exogenous
mol NADPH/min per lens with H2O2 or TBHP as the substrate. Measurements were performed at 37C.

carnosine entering the body intravenously, intraper-

itoneally, with food or topically to the eye is not
accumulated by the tissues, but is excreted in the
urine or destroyed by carnosinase, a dipeptidase
Conventional superoxide dismutase units per lens. Measurements were performed at 37C.
76.2 20.1

61.0 14.2

20.1 7.9
dismutased

present in plasma, the aqueous humour of the eye,

superoxide

liver, kidney and other tissues, except muscle and,

probably, the lens.[43,55-58] Among dipeptides of the
carnosine family tested as potential substrates for a
Lens homogenate

mol reduced NADPH/min per lens. Measurements were performed at 37C.

highly purified human serum carnosinase prepara-

tion, N-acetylcarnosine and a few other compounds
catalasec

7.0 1.8

6.7 1.1

6.0 3.4

were not hydrolysed,[57] thus promising a prolonga-

tion of physiological responses to therapeutic treat-
ments. A knowledge of corneal and iris/ciliary body
esterase activity, in particular acetylesterase and, in
Ratio of area of zone of opacity to total area of lens.

addition to esterase, the identified N-acetyltransfer-

decompositionb
Rate of H2O2

ase activities, prompted the development of L-

270 60

245 40

carnosine for ophthalmic application as an antioxi-

71 7**

dant, e.g. as the chemically characterised N-acety-

lated form of the dipeptide.[51] Because of its relative
mol H2O2/min per lens at 37C.

hydrophobicity compared with L-carnosine, N-

nmol H2O2/h per lens at 20C.

acetylcarnosine may gradually cross the cornea of

opacitya

0.10.7

0.81.0
Area of

the treated eye and maintain, over a longer period of

00.1

time, the concentration of active principle reaching

the aqueous humour. The naturally occurring com-
Transparent human

pound N-acetylcarnosine is proposed as an

Immature human

transparent lens.
cataract (n = 9)

cataract (n = 8)
Mature human

ophthalmic prodrug of L-carnosine that is resistant

mean SD)

to enzymatic hydrolysis by carnosinase.[51] Rabbit

(n = 10)

eyes treated with 1% N-acetylcarnosine, L-

Lens

carnosine or placebo and extracts of aqueous hu-

a
b

d
e
c

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)
136
mour from the anterior eye chamber were analysed
for imidazole content by reverse-phase analytical
high-performance liquid chromatography, thin layer
and ion-exchange chromatographic techniques.
Topical administration to the rabbit eye of pure
carnosine did not lead to accumulation of this com-
pound in the aqueous humor over 30 minutes in
concentrations exceeding that in the placebo-treated
matched eye.[51,52] N-acetylcarnosine showed dose-
dependent hydrolysis in its passage from the cornea
to the aqueous humour, releasing carnosine after
1530 minutes of ocular administration in a series of
therapeutic modalities: instillation subconjunc-
tival injection ultrasound-induced phoresis. Dif-
ferent treatment techniques showed excellent tolera-
bility of 1% N-acetylcarnosine by the animal and
human eye. Once in the aqueous humour, carnosine
might act as an antioxidant and enter the lens tissue
when present at effective concentrations
(515 mmol/L).[51]
7. Clinical Studies of N-Acetylcarnosine
for the Treatment of Cataracts
Two randomised, double-blind, placebo-control-
led trials of 6 and 24 months duration, respectively,
investigated a 1% aqueous solution of N-acetyl-
carnosine administered as two eyedrops twice daily.
A total of 49 elderly patients (average age 65 years)
with cataracts ranging in severity from minimal to
advanced (but not to the point of requiring surgery)
were treated; the total number of eyes affected was
76. Using a variety of sophisticated optical tech-
niques, the condition of the cataracts, visual acuity
and glare sensitivity were monitored.[46-48,59] All pa-
tients were evaluated at entry and followed up every
2 months for a 6-month period (trial 1) or at 6-month
intervals for a 2-year period (trial 2) for best-
corrected visual acuity and glare testing. In addition,
cataract was measured using stereocinematographic
slit images and retro-illumination examination of
the lens. Digital analysis of lens images displayed
light scattering and absorbing centres in two- and
three-dimensional scales.[59]
The eyes treated with N-acetylcarnosine were
substantially improved in 6 months: the measured
2004 Adis Data Information BV. All rights reserved.
Babizhayev et al.
transmissivity of the lenses increased in 42% of the
eyes by 1250%; in 90% of the eyes visual acuity
improved by 7100%; and in 89% of the eyes glare
sensitivity improved by 27100%. These improve-
ments were sustained for the duration of the
24-month trial. In no eyes was any worsening of the
condition seen. By contrast, the condition of the
untreated eyes in the control group worsened. Visual
acuity dropped in 89% of the controls by 1780%
after 24 months. The overall clinical results observ-
ed in the N-acetylcarnosine-treated group by the
24-month period of examination differed signifi-
cantly (p < 0.001) from the control group in the eyes
with cortical, posterior subcapsular, nuclear or com-
bined lens opacities.
Another interesting study by the same team also
evaluated patients between the ages of 48 and 60
years who had various degrees of sight impairment,
but who did not have the symptoms of cataract.[49]
After a course of treatment ranging from 2 to 6
months, it was concluded that the eyedrops alleviat-
ed eye tiredness and continued to improve sight (i.e.
there was more clear vision). The participants re-
ported that the treatment brightened and relaxed
their eyes. This is an important indicator that the
eyedrops have a value both for preventive purposes
and as medical applications as therapy.
Topical short- or long-term administration of 1%
N-acetylcarnosine to the eye was very well tolerat-
ed, with no ocular or systemic adverse effects, no
hyperaemia of conjunctival vessels, and no signs of
allergy or other toxic manifestations being reported.
All patients completed the study without any prob-
lems related to their allocated treatment.
8. Treatment of Age-Related Cataracts
in Canines
N-acetylcarnosine has been investigated in
canines with the aim of reversing cataracts in order
to avoid the need for surgery. The following
ophthalmic formulation (Can-C, Innovative
Vision Products, Inc., Delaware USA/International
Anti-Ageing Systems Group Ltd, Sark, Great Brit-
ain; Product is available from http://www.N-acetyl-
Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts
carnosine.com and http://www.can-c.net)1, which
includes N-acetylcarnosine, was used for the trial:
a
b
c ment was determined, and a new phenomenon of
Fig. 6. Reduction of cataract in canine eyes: (a) canine cataract
before treatment with 1% N-acetylcarnosine eyedrops; (b) 2 weeks
of treatment of the canine cataract with topical administration of 1%
N-acetylcarnosine eyedrops; and (c) results of the treatment after 1
month. Already the break-up of the impaired proteins is visible; an
effect that has been described as melting snow. The lens be-
comes clearer and the cortical opacities disappear partially on the
images.
1 The use of trade names is for product identification purposes only and does not imply endorsement.
2004 Adis Data Information BV. All rights reserved.
137
de-ionised water 970g; glycerine 1.0%, 13g; N-
acetylcarnosine 1.0%, 10g; carboxymethylcellulose
0.3%, 3g; benzyl alcohol 0.3%, 3g; potassium borate
7.91g (or the amount necessary to bring the solution
to about pH 6.36.5); and potassium bicarbonate
3.47g (or the amount necessary to bring the solution
to about pH 6.36.5).
This ophthalmic drug shows potential for the
nonsurgical treatment of age-related cataracts in
canines and has been shown to have high efficacy
and good tolerability.[49] Thirty dogs (50 eyes) were
allocated to the topical application of the drug in-
cluding 1% N-acetylcarnosine solution in eyedrops
twice daily to cure cataracts, and the control group
consisted of 15 dogs (40 eyes) that received placebo
eyedrops and ten dogs (20 eyes) that received no
eyedrops. The animal eyes were evaluated at entry
and followed up every 2 months for a 6-month
period. Cataract was measured using slit images and
retro-illumination examination of the lens. Coloured
analysis of the lens images displayed light scattering
and absorbing centres in the space scale. The overall
intra-reader reproducibility of cataract measure-
ments has been described earlier.[46,47,59] After 6
months, 96% of eyes treated with N-acetylcarnosine
in the described treatment formula of eyedrops
showed improvements in the slit image and retro-
illumination photographs of the lens. Typical results
of the study obtained at intervals of 1 month follow-
ing topical instillation of N-acetylcarnosine in
eyedrops are displayed in figure 6, using the retro-
illumination photography analysis technique. The
most striking results were obtained using a 1% N-
acetylcarnosine instillation in canines with age-re-
lated cataracts. The efficacy of the cataract treat-
melting snow was observed upon the instillation of
N-acetylcarnosine within only 1 month of long-term
treatment (see figures 6ac). The cortical appear-
ance of cataract reversal starts from the periphery
and then the lens becomes more transparent. This is
then accompanied by improved visual behaviour in
the animal.
Drugs R D 2004; 5 (3)
138
These results suggest that N-acetylcarnosine is
one of the most important endogenous antioxidants
for cataract prevention. N-acetylcarnosine and its
bioactivated principal carnosine appear to show
promise as water-soluble, universal antioxidants
that work at several levels to protect against oxida-
tive stresses to the lens and glycosylation and cross-
linking of the lens proteins, and protect the lens
proteins and membrane lipids from oxidative dam-
age, thus preventing and reversing age-related cata-
racts in human and canine eyes. A possibility exists
from our studies that carnosine is reacting directly
with malonyldialdehyde (MDA) and other al-
dehydes/ketones.[43] Indeed, carnosine has been
shown to protect against MDA-induced cross-link-
ing and toxicity, and a hydroxynonenal-carnosine
adduct has recently been characterised, providing
further evidence for the potential of carnosine as an
aldehyde scavenger.[60] The presented results can be
explained in part by the adduction of the various
LPO products directly by carnosine following
deacetylation of N-acetylcarnosine.
9. Conclusion
Senile cataract is a very common disorder, and
one must ask why many researchers have over-
looked the possibility of a nonsurgical approach to
treatment to date. The high costs of the surgical
procedure, the limited numbers of trained surgeons,
the potential risk of surgical complications, and the
simple fact that an artificial lens does not have the
optical qualities of a natural lens, make a nonsurgi-
cal procedure such as the use of N-acetylcarnosine
eyedrops a very attractive proposition. Application
of N-acetylcarnosine eyedrops appears to be a safe,
effective means to prevent cataracts, and possibly to
treat cataracts that are forming. In the future, this
type of therapeutic approach may provide another
option to cataract surgery and benefit the course of
cataract reversal and prevention in this age-related
eye disease.
Acknowledgements
This work was planned, organised and supported by Inno-
vative Vision Products, Inc., County of New Castle, Dela-
2004 Adis Data Information BV. All rights reserved.
Babizhayev et al.
ware, USA. Innovative Vision Products, Inc. is a holder of the
worldwide patent for the application of N-acetylcarnosine for
the treatment of ophthalmic disorders including cataracts.
References
1. Vision Research, a National Plan 1999-2002. Report of the
National Advisory Council. US Department of Health and
Human Services, Public Health Service, National Institutes of
Health, National Eye Institute 1998: 59
2. Kupfer C, Underwood B, Gillen T. Leading causes of visual
impairment worldwide. In: Albert DM, Jakobiec FA, editors.
Principles and practice of ophthalmology: Basic Sci. Philadel-
phia (PA): WB Saunders, 1994: 1249-55
3. Stark WJ, Sommer A, Smith RE. Changing trends in intraocular
lens implantation. Arch Ophthalmol 1989; 107: 1441-4
4. Kupfer C. The conquest of cataract, a global challenge. Trans
Ophthalmol Soc U K 1984; 104: 1-10
5. Gibson JM, Shaw DE, Rosenthal AR. Senile cataract and senile
macular degeneration: an investigation into possible risk fac-
tors. Trans Ophthalmol Soc U K 1986; 105 (Pt 4): 463-8
6. Harding JJ. Cataract: biochemistry, epidemiology and pharma-
cology. London: Chapman and Hall, 1991
7. Chatterjee A. Cataract in Punjab. Ciba Found Symp 1973; 19:
265-79
8. Tavani A, Negri E, La Vecchia C. Food and nutrient intake and
risk of cataract. Ann Epidemiol 1996 Jan; 6 (1): 41-6
9. Javitt JC, Tielsch JM, Canner JK, et al. National outcomes of
cataract extraction: increased risk of retinal complications
associated with Nd:YAG laser capsulotomy: the cataract pa-
tient outcomes research team. Ophthalmology 1992; 99:
1487-98
10. Javitt JC, Street DA, Tielsch JM, et al. National outcomes of
cataract extraction: retinal detachment and endophthalmitis
after outpatient cataract surgery: cataract patient outcomes
research team. Ophthalmology 1994; 101: 100-6
11. Canner JK, Javitt JD, McBean AM. National outcomes of
cataract extraction: III. Corneal edema and transplant follow-
ing inpatient surgery. Arch Ophthalmol 1992; 110: 1137-42
12. Hirsch RP, Schwartz B. Increased mortality among elderly
patients undergoing cataract extraction. Arch Ophthalmol
1983 Jul; 101 (7): 1034-7
13. Horwitz J, Jaffe NS. Anatomy and embryology. In: Podos SM,
Yanoff M, editors. Textbook of ophthalmology 3, lens and
cataract. New York: Gower Medical Publishing, 1992: 1.1-.9
14. Harding JJ, Crabbe MJ. The lens: development, proteins, meta-
bolism and cataract. In: Davson H, editor. The eye. Orlando
(FL): Academic Press, 1984: 207-492
15. Babizhayev MA, Nikolayev GN, Goryachev SN, et al. NMR
spin-echo studies of hydration properties of the molecular
chaperone alpha-crystallin in the bovine lens. Biochim Bi-
ophys Acta 2002 Jul 29; 1598 (1-2): 46-54
16. Benedek GB. Theory of transparency of the eye. Appl Opt 1971;
10: 459-73
17. Bettelheim FA, Siew EL. The effect of change in concentration
upon lens turbidity as predicted by the random fluctuation
theory. Biophys J 1983; 41: 29-33
18. Delaye M, Tardieu A. Short range order of crystallin proteins
accounts for eye lens transparency. Nature 1983; 302: 415-7
19. Bettelheim FA, Paunovic M. Light scattering of the normal
human lens: I. Application of random density and orientation
fluctuation theory. Biophys J 1979; 26: 85-100
Drugs R D 2004; 5 (3)
Lipid Peroxidation and Cataracts
20. Harding JJ. Cataract, Alzheimers disease, and other conforma-
tional diseases. Curr Opin Ophthalmol 1998; 9 (1): 10-13
21. Harding JJ, Dilley KJ. Structural proteins of the mammalian
lens: a review with emphasis on changes in development,
aging and cataract. Exp Eye Res 1976; 22: 1-73
22. Bloemendal H. Lens research: from protein to gene. Exp Eye
Res 1985; 41: 429-48
23. Spector A. The search for a solution to senile cataracts. Invest
Ophthalmol Vis Sci 1984; 25: 130-46
24. Bloemendal H. The lens proteins. In: Bloemendal H, editor.
Molecular and cellular biology of the lens. New York: John
Wiley, 1981: 1-47
25. Broekhuyse RM. Biochemistry of membranes. In: Duncan G,
editor. Mechnisms of cataract formation in the human lens.
New York: Academic Press, 1981: 151-91
26. Cotlier E, Obara Y, Toftness B. Cholesterol and phospholipids
in protein fractions of human lens and senile cataract. Biochim
Biophys Acta 1978; 530: 267-78
27. Babizhayev MA, Deyev AI, Linberg LF. Lipid peroxidation as a
possible cause of cataract. Mech Ageing Dev 1988; 44: 69-89
28. Halliwell B, Cross CE. Oxygen-derived species: their relation to
human disease and environmental stress. Environ Health Per-
spect 1994; 102 Suppl. 10: 5-12
29. Fecondo JV, Augusteyn RC. Superoxide dismutase, catalase
and glutathione peroxidase in the human cataractous lens. Exp
Eye Res 1983; 36: 15-23
30. Rao GN, Sadasivudu B, Cotlier E. Studies on glutathione S-
transferase, glutathione peroxidase and glutathione reductase
in human normal and cataractous lenses. Ophthalmic Res
1983; 15: 173-9
31. Babizhayev MA, Deyev AI, Chernikov AV. Peroxide-metabo-
lizing systems of the crystalline lens. Biochim Biophys Acta
1992; 1138: 11-9
32. Davies MJ, Truscott RJW. Photo-oxidation of proteins and its
role in cataractogenesis. J Photochem Photobiol B 2001; 63:
114-25
33. Spector A, Garner WH. Hydrogen peroxide and human cataract.
Exp Eye Res 1981; 33: 673-81
34. Palmquist BM, Philipson B, Barr PO. Nuclear cataract and
myopia during hyperbaric oxygen therapy. Br J Ophthalmol
1984; 68: 113-7
35. Bhuyan KC, Bhuyan DK, Katzin HM. Amizol-induced cataract
and inhibition of lens catalase in rabbit. Ophthalmic Res 1973;
5: 236-47
36. Koch HR, Beitzen R, Kremer F, et al. 8-Methoxypsoralen and
long ultraviolet effects on the rat lens: experiments with high
dosage. Graefes Arch Clin Exp Ophthalmol 1982; 218: 193-9
37. Bhuyan KC, Bhuyan DK, Podos SM. Free radical enhancer
xenobiotic is an inducer of cataract in rabbit. Free Radic Res
Commun 1991; 12-13: 609-20
38. Babizhayev MA. Accumulation of lipid peroxidation products
in human cataracts. Acta Ophthalmol (Copenh) 1989; 67:
281-7
39. Babizhayev MA. Lipid fluorophores of the human crystalline
lens with cataract. Graefes Arch Clin Exp Ophthalmol 1989;
227: 384-91
40. Babizhayev MA, Deyev AI. Free radical oxidation of lipids and
thiol groups in genesis of cataract. Biophysics 1986; 31:
119-25
41. Babizhayev MA, Linberg LF. Evidence of oxidation of the
polyunsaturated fatty acids in cataracts. Biochemistry (Mosc)
1986; 51: 1702-7
2004 Adis Data Information BV. All rights reserved.
139
42. Babizhayev MA, Deyev AI. Lens opacity induced by lipid
peroxidation products as a model of cataract associated with
retinal disease. Biochim Biophys Acta 1989; 1004: 124-33
43. Babizhayev MA. Antioxidant activity of L-carnosine, a natural
histidine-containing dipeptide in crystalline lens. Biochim Bi-
ophys Acta 1989; 1004: 363-71
44. Babizhayev MA. Failure to withstand oxidative stress induced
by phospholipid hydroperoxides as a possible cause of the lens
opacities in systemic diseases and ageing. Biochim Biophys
Acta 1996; 1315: 87-99
45. Babizhayev MA, Seguin NC, Gueyne J, et al. L-carnosine (-
alanyl-L-histidine) and carcinine (-alanylhistamine) act as
natural antioxidants with hydroxyl-radical-scavenging and lip-
id peroxidase activities. Biochem J 1994; 304: 509-16
46. Babizhayev MA, Deyev AI, Yermakova VN, et al. Efficacy of
N-acetylcarnosine in the treatment of cataracts. Drugs R D
2002; 3 (2): 87-103
47. Babizhayev MA, Deyev AI, Yermakova VN, et al. N-acetyl-
carnosine, a natural histidine-containing dipeptide, as a potent
ophthalmic drug in treatment of human cataracts. Peptides
2001; 22 (6): 979-94
48. Babizhayev MA, Yermakova VN, Deyev AI, et al. Imidazole-
containing peptidomimetic NACA as a potent drug for the
medicinal treatment of age-related cataract in humans. J Anti-
Aging Medicine 2000; 3: 43-62
49. Babizhayev MA. Method for topical treatment of eye disease
and composition and device for said treatment. US Patent
Application S.N. 10/480, 724, 2003
50. Halliwell B, Gutteridge JMC. Free radicals in biology and
medicine. Oxford: Clarendon, 1985
51. Babizhayev MA, Yermakova VN, Sakina NL, et al. N-acetyl-
carnosine is a prodrug of L-carnosine in ophthalmic applica-
tion as antioxidant. Clin Chim Acta 1996; 254: 1-21
52. Babizhayev MA, Yermakova VN, Semiletov YA, et al. The
natural histidine-containing dipeptide N-acetylcarnosine as an
antioxidant for ophthalmic use. Biochemistry (Mosc) 2000;
65: 588-98
53. Thomas JP, Girotti AW. Reactivity of photochemically-generat-
ed lipid hydroperoxides in cell membranes with glutathione
peroxidase. Photochem Photobiol 1989; 49: 153-6
54. Dahl TA, Midden WR, Hartman PE. Some prevalent bio-mole-
cules as defenses against singlet oxygen damage. Photochem
Photobiol 1988; 47: 357-62
55. Boldyrev AA, Dupin AM, Bunin AY, et al. The antioxidative
properties of carnosine, a natural histidine containing dipep-
tide. Biochem Int 1987; 15: 1105-13
56. Jay JL, Miller DJ, Morrison JD, et al. Histidyl derivatives in
rabbit lens and their diminution in human cataract [abstract]. J
Physiol (Lond) 1990; 420: 155
57. Jackson MC, Kucera CM, Lenney JF. Purification and proper-
ties of human serum carnosinase. Clin Chim Acta 1991; 196:
193-206
58. Lenney JF, Peppers SC, Kucera-Orallo CM, et al. Characteriza-
tion of human tissue carnosinase. Biochem J 1985; 228: 653-
60
59. Babizhayev MA, Deyev AI, Yermakova VN, et al. Image
analysis and glare sensitivity in human age-related cataracts.
Clin Exp Optom 2003; 86: 157-72
60. Aldini G, Carini M, Beretta G, et al. Carnosine is a quencher of
4-hydroxy-nonenal: through what mechanism of reaction?
Biochem Biophys Res Commun 2002; 298 (5): 699-706
Drugs R D 2004; 5 (3)
140 Babizhayev et al.

Correspondence and offprints: Dr Mark A. Babizhayev, In-

novative Vision Products Inc., Moscow Office, Ivanov-
skaya 20, Suite 74, Moscow 127434, Russia.
E-mail: markbabizhayev2004@yahoo.com

2004 Adis Data Information BV. All rights reserved. Drugs R D 2004; 5 (3)