SEARCH FOR NEW DRUGS

NYSTATIN:

METHODS OF PREPARATION, SEARCH FOR DERIVATIVES,

AND PROSPECTS FOR MEDICINAL USE (REVIEW)

Yu. D. Shenin, V. V. Belakhov,
and R. A. Araviiskii

UDC 615.332.012.071048.8]

The tetraene macrolide antibiotic nystatin was isolated in 1950 from a culture of
Streptomyces noursei, and immediately attracted the attention of investigators due to its
high antifungal activity [68, 69]. Nystatin has been produced in our country since 1962. The
antibiotic possesses a pronounced antimicrobial spectrum of activity, described in the USSR
State Pharmacopeia and the pharmacopeias of leading nations of the world, and is widely used
in medical practice. It is, prescribed for treating the most diverse manifestations of candidiasis, from thrush in newborns to systemic lesions in patients with malignant neoplasms,
weakened by the use of cytostatics.
The preparation is an amorphous or crystalline powder with a light-yellow color, practically insoluble in water and slightly soluble in polar organic solvents. Its solubility is
increased in aqueous-organic systems. In aqueous alcohols and acetone, the solubility of
nystatin passes through a maximum; the greatest solubility is achieved with a water content
of 30-40% [15]. In aqueous solutions, nystatin has a pK l and pK 2 of 2.64 and 9.73, respectively [46].
It has been shown that nystatin is a complex mixture of tetraene components, designated
Al, A2, and A s [43]. Different strains of Streptomyces noursei, depending on the genotype, can
synthesize preparations with different component ratios [99, i00]. For example, strain 153
produces nystatin consisting mainly of component A I [43], and strains LIA 0178 and 0187 with
A 2 and A 3 contents of 50 and 65%, respectively [41]. Thomas et al. [i00] have studied the
component composition of pharmacopeial forms of nystatin from different countries: the USSR,
China, Hungary, Italy, and the USA. While all forms met the requirements of the British pharmacopeia for physicochemical and biologic properties, the A I content of the mixtures ranged
from 3.8 to 78%. The component composition of nystatin produced by the Penzensk combine Biosintez has in recent years been stable, with an A I content not less than 90% and not more
than 10% nystatin A 2.
Physicochemical properties for nystatin components are given in Table I.
The components of nystatin differ from one another in their biological activity and
stability on storage [43].
A comparative study of therapeutic efficacy for sodium salts of components on generalized
candidiasis in white mice has shown that the sodium salt of nystatin A 3 is the most effective;
it also has the broadest therapeutic range. AI is considerably inferior in therapeutic activity to component A 3 [44].
The tetraene antibiotics polyfungin [75, 76, 88] and tetrafungin [103, 104] are similar
to nystatin. The first was isolated from a culture of Streptomyces noursei var. polyfungini,
the second from Streptomyces noursei subsp, tetrafungini. In contrast to nystatin, polyfungin
contains an additional component-polyfungin B [88, 89]. In tetrafungin, together with the
three components of nystatin, there is an original and predominant tetraene component [104].
It has also been shown that nystatin contains an original heptaene antibiotic, which we
have named nursimitzin [42]. Nursimitzin is generated during spontaneous oxidation of nystatin. The heptaene has also been demonstrated to be present in other nystatin preparations
[58, 83] and in tetrafungin [104].
More detailed study of its metabolic products has shown that Streptomyces produces the
nonpolyene antifungal antibiotics actiphenol, cycloheximide, and a number of other biologicAll-Union Scientific-Investigative Technologic Institute of Medicinal Enzymes and Antibiotics, St. Petersburg. Translated from Khimiko-farmatsevticheskii Zhurnal, Vol. 27, No.
2, pp. 14-21, February, 1993. Original article submitted November 15, 1991.

84

0091-150X/93/2702-0084512.50

9 1993 Plenum Publishing Corporation

TABLE i.
'Component
I ..

Physicochemical Properties of Nystatin Components

I (r (D~A)I

, Mol. wt.

Ai

C47H75Oi7
925

Az

C47HTsOI6

9 A3

909C~aHssO2o
1055

--16
,

+17,

~=36,o

Partition coefficient in system*

93
l
.I
2
1
' 2 9 1 , 3 0 4 , 318
" -(510, 83o, 795)
.. 290, 303, 318
(530, 740,

292, 3o5,~)

(405, 810; 760)

' 4,60 ~

2,88

2,47 .'"

16,8

1,85.

3,17

16,0
. . . . . .

*i) Amyl alcohol-isoamyl alcohol-citrate-phosphate

(2,5)
-.
4,85
3,54
~: (6,51 . . . . . . . . . . . . . . .

:}HPLC reten"Ition timer162

'37,4"
9

41,2

42,5

buffer pH 6.0 (12:17:29); 2)

MeOH-CHCI3-1% NaCI (2:2:1).

**Acetonitrile-acetate buffer pH 4.8 (Nucleosil 5 Cls).
ally inactive metabolites [61, 102]. Apparently, industrial strains have lost the ability
during selection to produce actiphenol and cycloheximide, and they have not been detected
recently in samples of nystatin.
Chemical structures have been established for nystatins At, A2, A3, and polyfungin B
[12, 50, 51,54, 72, 107].
For nystatin At, the absolute configuration of the asymmetric centers has been proven
[73, 77, 78, 90]. Its stereoselective synthesis has been carried out [84].
The molecules of components At, A2, A3, and polyfungin B contain an amino sugar, mycosamine (3-amino-3,6-dideoxy-D-mannose).
Its configuration was established by studying chemcal degradation products [106] and by electron diffraction of its hydrochloride [67]. There
is an additional deoxysugar in the nystatin A and polyfungin B molecules-digitoxose (2,6dideoxy-~-L-ribohexapyranose) [108]. In the macrocyclic components of nystatin there are
two chromophores, a tetraene and diene system of conjugated trans double bonds, and a large
number of hydroxyl groups. Therefore, the antibiotic molecule may be conventionally divided
into hydrophobic and hydrophilic parts. The hydrophobic part, by binding to sterols in biological membranes, forms an antibiotic-sterol complex, and the hydrophilic part forms an
aqueous pore within the membrane [ii]. The carbonyl group at C(13) and the hydroxyl at C(17)
form an internal hemiketal ring. Its presence in the nystatin molecule is confirmed by the
appearance, in the C 13 NMR spectrum, of a hemiketal carbon signal at 97.1 ppm [56, 86]. The
hemiketal ring has been shown to have a chair configuration [67]. The carboxyl and amino
groups endow the nystatin molecule with an amphoteric nature.
Fluorescence and circular dichroism spectra in ethanol, dimethylsulfoxide, and dimethylformamide solutions have revealed two forms of nystatin, characterized by different fluorescence times [37]. The ratio of these forms remained practically constant for all solvents
studied, over a wide temperature range ( f r o m - 1 9 6 to 70~
The authors suggest that they are
enantiomers, either in the amino sugar or hemiketal part of the molecule.
A number of investigators [i0, II, 49, 71, 81, 87, 91] have shown that the macrolactone
ring of nystatin is capable of considerable conformational rearransement, and may change from
a planar to a doubled-up form. These may explain the formation of nystatin preparations
that differ in physicochemical properties, under conditions of potentiometric titration and
photoirradiation.
Potentiometric titration curves of nystatin in water and organic solvents reveal three
main states:
inert (S), native (N), and activated (A). Preparations of all these states
have been isolated in solid form and their physicochemical characteristics determined [37,
38]. The most stable is S, in which form nystatin is isolated from the producing mycelium.
This is the state in which it is found in pharmaceuticals, and it is two orders less soluble
than N or A. When nystatin in state A is dissolved in methanol, it is spontaneously transformed to N, from which it may be photochemically converted to form A.
Shortcomings of the natural polyene macrolides, and in particular of nystatin, are insolubility in water and poor absorbability, which limits their sphere of use. A whole series
of nystatin derivatives has therefore been prepared. These may be divided into two groups:
ionic derivatives (salts and complexes) and solubilized preparations; and those with modifications to functional groups.

85

!-)--'k o . K

) "x/A~x/~xJm%/<~/'%//>'~J~'xN,/'~0g]C4s
:

/X/

".

~3

.....

/..~\ /R.,.../%/i,~,~,N.~\y/~/.~Vi.
2s

i)

E (:PolyfunginB ):.

X
OFi

,.-.

-~= ~

- ~ , A $ .

Polyfungin B ),

Nystatin ,\~-~i,~\5; Polyfungin B

.]
TrO~.

..~....

~
0

OH OE 0}{

<xd

OH ~i

~ ,::~" ~',k~"" , ~ " . / " , . . , / A ~ / ' , S ' % / / * v

'
~o--.._kS{

Component

:-\,

V - -

Among the first group should be noted the water-soluble sodium salts, hydrochlorides,
sulfates, and phosphates [9, 17, 57, 74, 92]. They are obtained by the addition of acids or
bases to methanolic solutions of the antibiotic, with subsequent precipitation by ether or
ethyl acetate [17]. Nystatin hydrochloride is soluble in water but rapidly loses activity,
which does not allow its use in medical practice. Aqueous solutions of the sodium salt of
nystatin have considerable viscosity; this has prevented its introduction as a pharmaceutical
preparation [17].
Methods have also been described for preparing nystatin salts with organic bases (for
example, N-methyl-D-glucosamine) [36]. Different investigators have shown that nystatin easily forms complex compounds with polyvalent ions, such as with cations [18, 19], complexes
containing the anion of an organic acid [20], with sodium deoxycholate [79], and with polyvinylpyrrolidone [12, 13]. The symmetry of nystatin's copper complexes has been determined.
It was found that nystatin may function as a uni-, bi-, and tetradentate ligand [105]. Complex compounds of nystatin with metal cations dissolve well in water and have antifungal
activity on a par with the starting antibiotic.
Complexes of nystatin with polyvinylpyrrolidone [13, 14], dispersed in water, form fine
suspensions over a wide pH interval. The authors chose system c o m p o s ~
for complete pre -~
cipitation of the resulting complex. It was demonstrated that solubilization is achieved
due to hydrogen bonds between hydroxyl groups of the antibiotic and keto groups of the polymer.
A whole series of derivatives of nystatin A z (I) have been prepared via the carboxyl
and amino groups (see scheme on next page).
In contrast to other polyene antibiotics, the acetyl and succinyl derivatives (II) of
nystatin were in fact biologically inactive [94]. However, when m-nitrobenzoyl chloride, or
chloroanhydrides of 5-nicotinic acids are used as acylalin,g agents, it is possible to
obtain N-substituted nystatin derivatives that display biologic activity equal to that
of the Starting preparation [39]. Nystatin reacts with aldose and ketonse mono- and
oligosaccharides under mild conditions with nearly quantitative yield [62-64]. With the use
of the Amadori rearrangement in dimethylformamide solution, in the presence of organic or
inorganic bases at 37~
N-substituted derivatives III are obtained. The compounds formed
are water-soluble and retain biological activity at the level of the starting nystatin,
against Saccharomycescerevisiae.
In addition, N-alkylsilyl derivatives IV of nystatin have been produced by reacting the
latter with trimthyl- and triethylchlorosilanes [8]. Organosilicon derivatives possess
marked antifungal activity. Their minimal fungistatic concentration against ii pathogenic
fungi in culture was 0.78-6.25 ~g/ml, and their acute toxicity was 3-4 fold less than the
starting antibiotic.
Hydrophosphoryl derivatives V [6] have been synthesized by reacting nystatin with aromatic aldelydes and hypophosphorous acid (Kabachnik-Fields reaction). The resulting compounds
had the same spectrum of antifungal activity, at the level of the starting compound, but the
toxicity was 2-3 fold lower.
86

Scheme for preparation of nystatin derivatives

o_.4~~ ~

I ~G

~o-k.ooo~

~" Y

c~_~~

~Z~o~ ~

I ~
o--~o~

,o..~x..co~ ~

(Ys~

~"v

c~

C~b

~~

.o--k~
O

%* " (
?

~
"

"

o~_

O_
~,

0-4"#3

~"~

p--ka~ ~

.o--<~ey-

"'6 Y
,

,,_

"~

~O~o~

o--~.~l
%

Y
l

~,
-

Among amino-group derivatives of nystatin, one should also note the amidines VI and enamines
VII, formed by reacting the antibiotic with acetylacteone ( ~ ) or acetoacetic acid ethyl ester
(for enamines), or with dimethylacetamide (DMA) (for amidines) at 20-35~ for 2 to 24 h, with
subsequent precipitation of the reaction product with ether [98]. The resulting derivatives
possessed better solubility in organic solvents and retained antifugal activity on a par with
the starting compound.
The methyl ester of N-(N',N'-dimethylaminoglycyl)nystatin VIII has been prepared. In
this case, nystatin reacts at 0~ in dimethylformamide solution with N,N'-dimethylglycine
hydrochloride, in the presence of triethylamine and phosphoric acid diphenyl ester azide
(DPAPA) [60]. The activity of the product was 2-fold less than that of nystatin.
The second functional group which has been used to obtain derivatives of nystatin is
the carboxyl group. When nystatin's caroboxyl group is esterified with diazoalkanes, esters
IX are produced, which retain antifungal activity comparable to the starting preparation [52,
53, 93]. The reaction is carried out in dimethylsulfoxide at 6~
the esters formed are precipitated with ethyl acetate. Transformation to the salt with acid via the free amino group
results in a preparation with high solubility in water at mildly acidic pH. While retaining
its antifungal spectrum of activity, nystatin methyl ester is 15 times less toxic.
However, a significant shortcoming of this method is its impracticability, in view of
the toxicity and explosion hazard of diazoalkanes. Therefore, the authors, in more recent
works, have proposed a method for methylating nystatin with methanol in the presence of
dicyclohexylcarbodiimide (DCC) at 30-40~ for 24 h [97].
On treating nystatin in alcoholic solution with dimethylsulfate in the presence of sodium
carbonate as a neutralizing agent, one obtains the methyl ester of nystatin trimethylammonium
salt X [65, 66]. Its biologic activity was 2.5-fold lower than that of the starting material,
but its toxicity was decreased 10-fold. The resulting salt was soluble in water at concentrations up to 10%.
Aliphatic amides of nystatin XI have been synthesized. The reaction was carried out in
dimethylsulfoxide at room temperature with the corresponding amine, in the presence of triethylamine and DPAPA [31]. The amides formed water-soluble salts with inorganic and organic
acids, the antifungal activity of which was similar to that of the starting nystatin.
A new type of nystatin derivative via the carboxyl group is X!I, which is obtained by
reacting the DPAPA-activated carboxyl with l-amino-4-methylpiperazine in dimethylformamide at

87

TABLE 2. Antifungal Activity Spectrum for Nystatin (complex) and Its Components,
and for Polyfungin B
Test culture

I
Candida albicans
Candida tropicalis "
Candida pseudoiropiealis
Candida krusei
Candida parapsilosie
Candida stellatoidea
Candida quiltermondii
Candida utilis
Saccharomyces eerevisiae
Torulopsis glabr,~ta
Cryptococcus neoformans
Histoplasma capsuiatumCoccidiodes immitis
Sporotrichum s c h e n e k i i
Phialophora verrucosa
Aspergillus ftimigatus :
Trichophyton rubrum
.Trichophyton men-tagrophytes
Microsporum gypseam
Prototheca filamenta
Blastomyces dermatitidis
Hormodendrum pedrosoi.,

complex
5,9
6,9
4,7
7,4
7,2
4,0
4,0

Minimal fungistatic concentration

p01yfungin B "
A,
I
A~

(1,2--10,0)
(2,0--10,0)
(4,1--5,0).
(4,1--6,1)
(3,1--IO,O)

1,57
1,57

0,78
0,78

0,78

1,57

1,57

1,57

1,57

0,78

1,57

(2,o--5,o)

(2,0-- I0,0)

],6

2,3 (1,6--3,0)
3,1
1,56 ( 1,5-- i,6)
1,56.
0,52 (0,2--i,s6)
9,2 (6,0-- 12,5)
12,5
7,6 (3,2--12,0)
9,5 {6,2--12,5)
9,2 (6,0--12,5)
20,0

6,25
3,14 ,

1,57
1,57

10,0

. .10,0

1,57
3,14

!,57
3,14

0,78

3,14
1,57
10,0 .

1,75(0,56--3,12)
1,56

1,57

6,25

0 ~ in the presence of triethylamine [59]. Hydrazides of salts, formed with organic and inorganic acids, had biological activity close to that of nystatin, and were well soluble in
water.
For extracting nystatin from wet mycelium, the lower alcohols, acetone, 80% acetic acid,
pyridine, dimethylformamide, and other organic solvents have been used [i, 2, 28, 32, 34, 35].
In [29] methanolic solutions of sodium hydroxide or ammonia, or 3% solutions of oxalic or
citric acids in methanol were used to extract nystatin from mycelium. The method is based
on the improved solubility of the antibiotic in methanol at acidic or alkaline pH values.
Nystatin is then isolated by adding water to the extracts and adjusting to neutral pH.
Methanolic solutions of mineral acids (for example, sulfuric acid at pH 3.0) or amines,
ammonia at pH 9.6, or buffers with solubilizing agents may also be used for extraction [22,
24]. The solubilizing agents can be bases, calcium and magnesium salts of cyclohexylsalicylic
acid, 2-hydroxycyclohexylbenzoylglycine, anion-active, sodium or calcium salts of saccharin,
and benzyl alcohol [25].
Dry mycelium is extracted with a 2% solution of CaCI 2 in methanol, followed by vacuum
concentration of the solution and decomposition of the CaCI 2 complex with water [3, 5, 38].
The crude antibiotic is desorbed by treating the aqueous suspension with hydrogen peroxide [30], which decomposes impurities and increases the biologic activity of nystatin by 800i000 units, or by treating with aqueous sulfur or sulfite solutions at pH 4.5-5.0 at 50 ~ , or
passing SO 2 through the aqueous suspension [27].
For purifying nystatin, the following methods are used: partitioning the antibiotic in
diphasic systems containing butanol, methanol, hexane, and water [26]; reprecipitating nystatin as the CaCl= complex [4]; extracting crude material with acetone, saturated sodium iodide
or sodium, potassium, or ammonium thiocyanate, with subsequent precipitation of the antibiotic
with water [23]; using ion-exchangers, in particular XAD-2 resin [33]; and crystallization
from aqueous solutions of lower alcohols [21]. Crystalline nystatin is thus obtained.
Raising the temperature in the presence of moisture and atmospheric oxygen, as well as
daylight, during isolation and purification of the antibiotic leads to rapid destruction of
the tetraene chromophore. Various methods are used to counteract these factors (shortening
the time of technical operations, carrying out the process in an atmosphere of inert gas
and at low temperatures, using complex-formers and antioxidants, and other methods).
A unified method for isolation and chemical purification of polyene antibiotics, including
ing nystatin, has been developed, calling for combined use of the antioxidant ionol, inert
gases (nitrogen or argon), and complex-forming agents (Triton B, tripolyphosphate), which prevent oxidative inactivation of preparations [7, 45].

88

Nystatin has a wide spectrum of antifungal activity (Table 2).

It is important to note that strains of pathogenic fungi, isolated from patients, may
differ significantly in sensitivity [48].
Resistance to nystatin in vitro develops quite slowly, after multiple passages. In this
regard, cross-resistance to other polyene antibiotics is also seen. Acute toxicity of nystatin for laboratory animals (LDs0) is equal to 30 mg/kg intravenously, 45 mg/kg intraperitoneally, and by mouth - more than 8 g/kg. The antibiotic is strongly bound to serum proteins.
With oral use, nystatin is excreted in the feces almost entirely unchanged. Even when i0
million units are given, only traces are found in the blood, without antifungal effect.
In the first experiments with nystatin, mice were infected intraperitoneally with ~.
albicans and were given chlortetracycline, to amplify the infection. All animals died within
a few days. A single dose of nystatin (3 mg), given subcutaneously 2 h prior to or simultaneously with infection, protected two-thirds of the animals from death up to day 16.
In many subsequent studies, a positive effect of nystatin on survival of laboratory animals was demonstrated with subcutaneous, intravenous, intramuscular, and oral use. However,
patholomorphologic changes and seeding by fungi of internal organs persisted. Experiments
were done with the use of nystatin to treat deep mycoses. In [55], its chemotherapeutic efficacy in experimental coccidioidsis was studied. With a 10-day course of treatment with intravenous nystatin (i mg/mouse, daily), after two weeks all the animals were alive. In the control, untreated group, 35% of animals died. However, deaths were eventually observed among
treated animals, with a 22.5% fatality rate at day 28 and 31.5% at day 40. In the control
group at these times, the death rate reached 50 and 87.5%. Tests of intravenous nystatin on
5 patients with cavernous histoplasmosis and coccidiomycosis were unsuccessful; due to a
large number of side effects, treatment in 4 patients was discontinued [80]. One patient with
histoplasmosis did receive treatment (200,000 units per day) up to day 62, but without apparent effect.
In clinical practice at present, nystatin is used orally and locally, mainly to treat
candidal lesions localized to the skin and mucous membranes. Oral candidiasis in newborns
and children is usually cured within 3-15 days [47].
Despite the appearance in medical practice of modern synthetic antibiotics, nystatin
has not lost its importance, and is used for candidal complications of HIV infection. When
treating candidal stomatitis, glossitis, and esophagitis in AIDS patients, nystatin has been
used successfully as a suspension of 250,000-500,000 units, every 4 h for 2 weeks [95].
Response to the medication is good; however, frequent recurrences are noted. In connection
with this, such patients are begun on treatment with ketoconazole (Nizoral) 200-400 mg every
12 h, and then over time are placed on nystatin (in the usual doses) as supportive therapy.
Nystatin suppositories, tablets, and ointments are used to treat candidiasis of the
female reproductive tract. For treating urogenital candidates, nystatin tablets are prescribed to be taken at 4 million units per day for 2 weeks, with simultaneous local application of nystatin ointment [16, 40, 85]. Results of treating generalized candidiasis with
nystatin do not always support its suitability for this, although the cutaneous manifestations
frequently regress. There is a report on curing candidal peritonitis with nystatin [70].
Inhalation of nystatin suspension has a pronounced clinical effect on pulmonary aspergillosis
[96].
For prophylaxis of candidiasis, which often accompanies prolonged use of broad-spectrum
antibacterial antibiotics, preparations containing nystatin in combination with antibiotics
are often utilized.
Nystatin enters into the scheme of combination therapy for cancer patients receiving
cytostatic preparations [i01].
For preventing fungal endocarditis, which complicates open-heart surgery, 1.5 to 3 million units of nystatin is prescribed daily as tablets for 4-6 weeks [i01].
Nystatin is produced in many countries in the form of tablets, suppositories, beads,
and ointments. To protect against inactivation by gastric juice, tablets are coated with a
protective membrane.
Nystatin does not cause serious side-effects, even with prolonged internal use. With
large doses there may be gastrointestinal disturbances noted - nausea, vomiting, lower bowel
upsets. Decreasing the dose lessens these side-effects.
89

In our country, nystatin is used clinically in the following pharmaceutical forms:

coated tablets of 250,000 and 500,000 units (VFS 42-1684-87); ointment, I00,000 units/g (FS
42-1271-87); vaginal suppositories, 250,000 and 500,000 units (FS 42-2017-83); suppositories,
2,500,000 and 500,000 units (FS 42-2016-83); coated tablets of tetracycline with nystatin,
i00,000 units (FS-42-36-72); nystatin for veterinary use (TU-64-3-223-85).
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2.
3.
4.
5.
6.
7.

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90

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ACTIVITY OF CINNAMIC ACID DERIVATIVES AND NEW METHODS FOR

THEIR SYNTHESIS (REVIEW)
A. V. Simonyan

UDC 615.31:547.586.5].015

Derivatives of cinnamic acid, displaying a broad spectrum of biological activity and low
toxicity, are of definite interest for the purposes of creating new highly effective drugs
based on them.
In this report, we attempt to systematize data on the biological activity of cinnamic
acids substituted on the phenyl radical and a-substituted cinnamic acids, and to show the
connection between the different and most characteristic types of biological activity.
Many pathological states of living organisms due to hypoxia of various etiologies, toxic
effects, inflammatory processes, ischemia, stress, etc. are connected with damage to the cell
membranes.
One universal mechanism for cell damage is free-radical oxidation of their membrane lipids [12, 33]. On this basis, we can assume that compounds displaying antioxidant
and membrane-stabilizing activ%ty may prevent development of a whole series of pathological
states.
Accordingly, for preliminary screening of pharmacologically active compounds, often we
resort to in vitro determination of the antioxidant and membrane-stabilizing activities,
which makes it possible to significantly shorten the process of selection of the most active
structures.
However, we should note that even when in vitro investigation of the qualitative
and quantitative characteristics of the structural-biological activity relation gives satisfactory results, the determined characteristics are not always confirmed by in vitro experiments, since in practice it is impossible to completely reproduce the conditions of the living organism. However, investigations determining the structure vs. activity characteristics
make a definite contribution to the development of goal-directed synthesis of biologically
active compounds.
i_. Antihypoxic, Hepatoprotective, and Anti-inflammatory Activity. The Approach described
above was used in studying the structure-activity relation for derivatives of 4-alkoxy-substituted cinnamic acids. As a result of in vitro investigations, it had been established that
the membrane-stabilizing activity of the compounds increases with an increase in the positive
electronic effect of the substituents in the aromatic nucelus and their lipophilicity. At
the same time, a positive inductive effect of the ~-substituents reduces the membrane-stabilizing activity.
Introduction of the lipophilic alkoxy group in the 4 position combined with
introduction of a halogen at the 3 position, increasing the overall acidity of the molecule,
leads to enhancement of the membrane-stabilizing activity; this has been confirmed by synthesis of new compounds considering this prediction.
However, an attempt to transfer the determined characteristic to in vivo experiments in a study of the anti-inflammatory activity of
the compounds obtained did not lead to the expected results [51].
For derivatives of hydrocinnamic acid (characterized by high antioxidant activity), antiinflammatory, analogesic, and antipyretic activity has been described. Thus drugs have been
proposed based on 4-cyclohexyl-g-[(alkyl)alkanoylthio]hydrocinnamic acid which display the
above-indicated types of activity [17]. Furthermore, drugs have been proposed which have
Pyatigorsk Pharmaceutical Institue. Translated from Khimiko-farmatsevticheskii Zhurnal,
Vol. 27, No. 2, pp. 21-27, February, 1993. Original article submitted April 9, 1991.
92

0091-150X/93/2702-0092512.50

9 1993 Plenum Publishing Corporation