CC BY
13Chemical Journal of Kazakhstan
ISSN 1813-1107, elSSN 2710-1185 https://doi.org/10.51580/2021-1/2710-1185.31
Volume 2, Number 74 (2021), 87 - 103
УДК 615.017:616.079
STRUCTURE AND PROPERTIES OF THE DI-((2S)-2-AMINO-3-(1H-INDOL-3-YL)PROPIONATE)DIHYDROTETRAIODIDE
A.N. Sabitov , S. Turganbay, A. Dzhumagazieva
Test Facility, JSC «Scientific center for anti-infectious drugs», Almaty, Kazakhstan
*E-mail: aitugans@mail.ru
Abstract: New organic iodine complex in the amino acid - alkali metal salt - iodine -water system was synthesized. The physico-chemical properties of complex di-((2S)-2-amino-3-(1H-indol-3-yl)propionate)dihydrotetraiodide were studied. Microscopic analysis shows that particles of complex have elongated needle-like linear stick-like shapes with average size 2.50-4.00 ^m. The cytotoxicity test on MDCK cell culture, antimicrobial activity on S. aureus ATCC 6538-Р (museum susceptible strain); S. aureus ATCC-BAA-39 (museum multiresistant strain); E. coli ATCC 8739 (museum susceptible strain); E. coli ATCC-BAA-196 (museum multiresistant strain); P. aeruginosa ATCC 9027 (museum susceptible strain); P. aeruginosa TA2were observed. Complex has low cytotoxicity, direct antiviral effect, and antimicrobial activity against antibiotic-resistant strains of microorganisms. Di-((2S)-2-amino-3-(1H-indol-3-yl) propionate)-dihydro-tetraiodide does not cause any mutagenic effect on mammalian cells of the L5178Y line, both in the presence and in the absence of metabolic activation.
Keywords: iodine polyanion complex, Di- ((2S) -2-amino-3- (1H-indol-3-yl) pro-pionate)-dihydro-tetraiodide, x-ray diffraction analysis, mutagenic test on L5178Y line, cytotoxicity.
1. Introduction
Semiorganic compounds, including metal atoms and various organic molecules are of great interest from the point of view of the diversity of their pharmaceutical and physical properties. For example, the large glycine family of metal halides and metal halides includes a large variety of different crystal structures. The structural units of these compounds are glycine molecules, halide ions, and metal cations. Depending on the combination of building blocks, five different structural families of glycine halides and glycine metal halides were distinguished [1]. Studies of the physical properties have shown that some of
Citation: Sabitov A.N., Turganbay S., Dzhumagazieva A. Structure and properties of the di-((2S)-2-amino-3-(1H-indol-3-yl)propionate)dihydrotetraiodide. Chem. J. Kaz., 2021, 2(74), 87-103. DOI: https://doi.org/10.51580/2021-1/2710-1185.31
these compounds are ferroelectrics, ionic conductors, and exhibit interesting thermal and optical properties [2-9]. Due to the presence of asymmetric centers in most amino acid molecules, coordination compounds with amino acids crystallize within non-centrosymmetric spatial groups, being possible candidates for nonlinear optical materials and are interesting compounds because of their possible use in new optoelectronic technologies [10, 11].
In our early studies, we have already studied semi-organic iodine complexes in the amino acid - alkali metal salt - iodine - water (or organic solvent) system. A number of them had significant biological activity against pathogenic microorganisms and viruses [12-14].
The aim of this research is the synthesis of new compound in the tryptophan-iodine-sodium iodide-water system, determination of the crystal structure of new compound and physico-chemical properties of the obtained complex.
2. Experimental part
Synthesis of new semiorganic iodine complex. 2.000 g of tryptophan, 11 mL of purified water, and 3 ml of hydrochloric acid (18%) were added to a 50 mL beaker. The mixture was stirred with a glass rod until complete dissolution of the amino acid, while heating the beaker on a water bath. The molar ratio of tryptophan: NaI : I2 was 1:0.5:1. 0.7340 g of sodium iodide, 1.0132 g of iodine, previously crushed in an agate mortar to a fine powder, and 20 mL of ethanol (96%) were added to a 100 mL flask with ground-in stopper. The mixture was stirred until complete dissolution, while heating on a water bath. After complete dissolution of the mixture in the flask the sodium triiodide solution was added to the tryptophan solution. The resulting reaction mixture was capped and stirred at room temperature for 5 min. The prepared product was kept in a dark place at room temperature.
A day later the complex solution was poured into a crystallizer, a crystallizer with the complex solution was further placed into a desiccator with anhydrous calcium chloride to evaporate the solvent at room temperature. After complete evaporation of the complex solution we took the monocrystals of the new tryptophan: NaI:iodine system and the yield of monocrystals of the complex was 1.95 g (65% of the theoretical).
X-ray diffraction analysis. The purpose of these studies was to determine and refine the crystal structure of the new coordination compound formed in the tryptophan: NaI:iodine:water system. Small crystals were formed after crystallization of the synthesized compound. All diffraction measurements were carried out by using Enraf-Nonius CAD4 autodiffractometer (Nederland) (graphite monochromator, Mo-Ka radiation, 9/29 scan, CAD4 software) [15]. The main fragment of the structure was decoded by a direct method. The coordinates of missing atoms were determined by difference Fourier syntheses of electron density. Due to the presence of heavy atoms in the structure, it was not possible to detect hydrogen atoms by difference Fourier syntheses of electron density.
The coordinates of hydrogen atoms were determined by geometric calculations and refined using the "rider" model with the following conditions: C-H bond length = 0.96A, Uiso(H) =1.5Ueq (C) for CH2 groups, CH = 0.93A, Uiso (H) =1.5Ueq (C) for CH groups, NH = 0.86A, Uiso (H) = 1.2Ueq (N) for NH groups, NH = 0.89A, Uiso (H) = 1.2U«j (N) for NH3 groups and O- H = 0.86A, Uiso (H) = 1.2Ueq (O) for water molecules. The structure was refined using full matrix OLS in the anisotropic approximation for non-hydrogen atoms and in the isotropic approximation for hydrogen atoms. All structural calculations were carried out using the SHELXTL [16] and JANA2006 [17] software packages; the purity of structural data was checked using the PLATON software [18].
Investigation of physical-chemical parameters. UV-VIS spectra of the water solution of obtained compound was measured using an UV-Vis spectrophotometer Perkin Elmer Lambda-35 at wavelength from 200 to 900 nm.
Determination of the melting point of the compoundwere carried out using a synchronous thermoanalyzer STA 449 F1 Jupiter (NETZSCH) combining simultaneous measurement of mass changes (thermogravimetry) and heat fluxes. Samples weighing 0.8-2 mg were heated in the temperature range from 28 to 320 °C in 85 ^l closed corundum crucibles at a rate of 10 K/min in a dry nitrogen atmosphere (gas flow rate 40 ml/min). The thermograms were recorded differential curves of change in the heat capacity of the sample.
The determination of the melting temperature of substances was carried out by using the Netzsch-Proteus program at the intersection point of the tangents held to the beginning of the endothermic peak on the Differential Scanning Calorimetry (DSC) curve. The temperature of the beginning of the peak, which corresponds to the melting point of the sample, was determined.
Bulk density (kg/m3) is determined by weighing a graduated cylinder filled with a substance after repeated compaction. Particle shape and size were determined using a Nikon Eclipse 50 i microscope and NIS Elements AR software. Solubility is determined in purified water, as well as several polar and non-polar solvents. The measurement of pH is carried out by potentiometric method using a combined electrode on an ionometer Basic pH Meter PB-11, manufactured by Sartorius. The test was carried out on 1% aqueous solutions of the substance.
Determination of cytotoxicity. Cytotoxicity is the indicator of the quality of being toxic to cells. The in vitro cytotoxicity of substances was determined using the MTT test. Cells were seeded into 96-well plates at a concentration of 200,000 cells in 1 ml. Dies were cultivated in a thermostat at 37 °C, 5% CO2. After 24 h of incubation, the growth medium was removed from the wells of the plate, and 200 ^l of the medium containing the test substances was added. 200 ^l DMEM nutrient medium was added to the negative control wells.
After 48 and 72 h, the medium with the substance was removed from the wells, 200 ^l of fresh nutrient medium and 50 ^l of MTT working solution (5 mg/ml) were added, the plate was incubated for 4 hours at 37 °C. After the expiration of the incubation period, the supernatant was removed. 100 ^l of DMSO was added to each well. The optical density in the wells was measured on
a Tecan Sunrise RC.4 microplate reader, Austria, at the wavelength of the main filter - 492 nm and the reference filter - 620 nm. The calculation of the results was performed using formulas 5-7:
- calculated the arithmetic average value of optical density (Y) for the negative control by theequation (5):
F=y-+-±y,=I£y, (5)
n n~~{
Where, yi - measurement result of the optical density (OD) of each object of the group; n - is the number of objects in the group; - calculated the percentage of surviving cells for each repetition of each concentration of the test substance according to the (6):
Y
Percentage of surviving cells = -=^ *100%. (6)
YNC
Where, Yi - the result of the measurement of the OD for each group; Y NC -
arithmetic average OD (Y ) for negative control; - calculated the arithmetic
average value of the percentage of surviving cells (Y ) for each concentration of the analyte by the formula (3);
- CTC50 (concentration of substances at which 50% of cells die) for each test substance was calculated as in (7):
CTC = ° 1
X1 50 -* (Mx 2 - Mx1)
X1 - X 2
+Mx1. (7)
Where X1 - > 50 % surviving cells; X 2 - < 50 % surviving cells;Mx1- concentration of matter where survived > 50 %; Mx2 - concentration of matter where survived < 50 %.
Studies on antimicrobial activity. The antimicrobial activity of the coordination compound was studied by using the twofold serial dilution method in a liquid nutrient medium [19].
The procedure for antimicrobial activity was performed by the method of twofold serial dilutions in a liquid nutrient medium (CLSI M100, 2016). For the method of two-fold serial dilutions, an inoculum of the test strain of the microorganism at a concentration of 1.5 x106 CFU/ml was used. The primary suspension of the test strain was prepared in a physiological solution (0.9% NaCl). An aliquot of a day-cultured test strain was taken with a sterile loop. After that it was introduced into a sterile tube with 5 ml of 0.9% NaCl. The turbidity of the inoculum obtained was monitored by measuring the optical density on a DEN-1 densitometer (Biosan, Latvia). The density of the primary suspension was 0.5 units. According to McFarland, which corresponds to 1.5 x 108 CFU/ml. Next, the primary suspension in an amount of 0.1 ml was introduced into a test tube with 9.9 ml of isotonic solution to achieve a working
concentration of 1.5 * 106 CFU/ml.Testing was carried out on a liquid nutrient medium - Muller-Hinton broth (Himedia, India).
A 48-well plate (BIOLOGIX, China) was used to determine the antimicrobial activity. In all the wells, except the 1st (from 2 to 16), poured nutrient broth Muller-Hinton (MHB) in an amount of 0.5 ml. Working solutions of coordination compounds were introduced in a volume of 0.5 ml into the 1st tube, and the second with MHB already present in it (0.5 ml). Next, serial dilutions were made, which were carried out by taking the mixture (MHB (0.5 ml) + test compound (0.5 ml)) from the 2nd tube in the amount of 0.5 ml to the 3rd tube, which already contains 0, 5 ml of broth, etc. From the last tube, 0.5 ml of the mixture was removed. Thus, the following dilutions were obtained: 1: 0; 1: 1; 1: 2; 1: 4; 1: 8; 1:16; 1:32; 1:64; 1: 128; 1: 256; 1: 512; 1: 1024; 1: 2048; 1: 4096; 1: 8192; 1: 16384, which corresponds to the tubes from the 1st to the 16th. The 17th test tube was the control of culture growth.
After conducting a series of dilutions, 0.05 ml of microorganism test strain was added to all tubes at a concentration of 1.5*106 CFU/ml. The procedure was repeated for all test cultures. All samples were incubated for 18-24 hours at a temperature of (37±1) °C. After the time of incubation, seeding was carried out on a dense nutrient medium - Muller-Hinton agar (Himedia, India) to determine viable cells. After seeding the cups were placed in a thermostat for 18-24 h, the cultivation was carried out at a temperature of (37±1) °C.
The results were recorded by the presence / absence of visible growth of microorganisms on the surface of a dense nutrient medium. The minimum bactericidal concentration (MBC) was considered the lowest concentration in the well, which completely suppressed the growth of microorganisms. All experiments were performed in triplicate.
Test strains used in the study were obtained from the American Type Culture Collection (ATCC). The museum susceptible, museum multiresistant, and one clinical test strains were used in the experiment: S. aureus ATCC 6538-P (museum susceptible strain); S. aureus ATCC-BAA-39 (museum multiresistant strain); E. coli ATCC 8739 (museum susceptible strain); E. coli ATCC-BAA-196 (museum multiresistantstrain); P. aeruginosa ATCC 9027 (museum susceptible strain); P. aeruginosa TA2 (clinical multiresistant strain) [20].
Investigation of mutagenic properties in in vitro micronucleus assay. The choice of the investigated concentrations of the complexes based on the range of concentrations of drugs recommended for their study on the putative mutagenic activity in vitro. According to the OECD Guideline for Research on Chemical Substances No. 487 "In vitro micronucleus test of mammalian cells", adopted on September 26, 2014 [21], for non-toxic, highly soluble compounds, the maximum dose can be up to 5 mg/ml,
The suspension culture of L5178Y cells is recommended for MNvit by the OECD Guidelines for Testing Chemical Substances No. 487.
Murine L5178Y lymphoma cells are used because they are sensitive indicators of mutagenic activity of a wide range of chemical compounds.
3. Results and Discussion
Structure of the obtained crystals. The resulting organic complex is a dark gray-green with a weak iodine odor (Figure 1). Particles have elongated needlelike linear stick-like shapes. All synthesized amount of the monocrystals of new compound were stored at dark place for further use. The parameters of triclinic unit cell were determined (Table 1) and refined based on 25 reflexes with 10.2 < 0 < 12.9.
Figure 1- Photo (a) of the obtained di-((2S)-2-amino-3-(1H-indol-3-yl)propionate)-dihydrotetraiodide complex and (b) its particles sizes.
Table 1 - Crystallographic data for C„H12N202KC„H13N202)+Kl4)2-^NaMH20, determination accuracy is given in parentheses
Crystallographic data
Formula C11H12N2O2^(C11H13N2O2)+^ Na+-2H2O- (I4)-2
Molecular weight 976.08
Syngony; Space group Triclinic, Р1
Lattice parameters a, b, c [A] 5.4298(11), 11.427(2), 13.388(3)
alpha, beta, gamma [deg] 101.58(3), 100.96(3), 93.43(3)
V [A]3; Z 794.8(3); 1
D(calc)[g/cm3]; F(000) 2.039; 460
Mu (MoKa) [ mm-1 ] 3.973
Crystalsizes [mm] 0.22x0.10x0.005
Measurements
Temperature (K); Radiation+ [A] 293; MoKa; X=0.71073
0min;0max[Deg] 1.6, 30.0
Measuring area -7<h<7; -16<k<16; -18<l<18
Number of reflexes 7080
Observed data [I > 2.0 sigma(I)] 2408
Refinement
Number of reflexes, Number of parameters 7080, 363
R, wR2, S 0.0784, 0.2176, 0.97
w = 1/[\sA2A(FoA2A)+(0.0349P)A2A+8.8867P] where P=(FoA2A+2FcA2A)/3
Max. and Av. Shift/Error 0.00, 0.00
Min. and Max. Resd. Dens. [e/A3] -0.45, 0.41
According to the results of the calculations performed, the chemical structure of the compound can be represented as CnH12N202^(CnH13N202)+^Na^2H20^(I4) 2-. By IUPAC it will be di-((2S)-2-amino-3-(1H-indol-3-yl) propionate)-dihydro-tetraiodide. Interatomic distances in the crystal structure are given in Table 2.
Table 2 - Main interatomic distances in the structure C„H12N202KC„ H13N202)+^Na+^2H20^(I4) 2-
Atoms Distance (Â) Atoms Distance (Â) Atoms Distance (Â)
I1-I2 2.762(4) C4-C5 1.38(5) C1-C2 1.44(4)
I1-I3A 3.431(4) I1-I3B 3.444(4) C1-C6 1.41(3)
I2-I4B 3.628(4) I2-I4A 3.629(4) C1-C9 1.42(5)
Na1-01W 2.30(2) C5-C6 1.35(4) C2-C3 1.41(5)
Na1-02W 2.36(3) N7-H7 0.8600 C3-C4 1.40(5)
Na1-035 2.43(2) C8-C9 1.41(4) C5-H5 0.9300
Na1-014_b 2.39(2) C9-C10 1.45(4) C8-H8 0.9300
Na1-015 2.42(2) C10-C11 1.55(3) C10-H10A 0.9700
Na1-034_b 2.34(3) C11-C13 1.51(3) C10-H10B 0.9800
014-C13 1.29(3) N12-H12B 0.8900 C11-H11 0.9800
015-C13 1.21(3) N12-H12C 0.8900 C22-H22 0.9300
034-C33 1.35(4) N12-H12A 0.8900 C23-H23 0.9300
035-C33 1.17(4) C21-C22 1.34(4) N32-H32C 0.8900
01W-H1W2 0.87(11) C21-C26 1.49(4) N32-H32B 0.8900
01W-H1W1 0.86(7) C21-C29 1.40(5) C2-H2 0.9300
02W-H2W1 0.86(17) C22-C23 1.31(4) C3-H3 0.9300
02W-H2W2 0.86(6) C23-C24 1.47(5) C4-H4 0.9300
N7-C6 1.37(3) C24-C25 1.30(5) C24-H24 0.9300
N7-C8 1.39(4) C25-C26 1.37(4) C25-H25 0.9300
N12-C11 1.43(3) N27-H27 0.8600 C28-H28 0.9200
014-H14 0.8200 C28-C29 1.37(4) C30-H30B 0.9700
N27-C28 1.34(4) C29-C30 1.50(4) C30-H30A 0.9700
N27-C26 1.36(4) C30-C31 1.52(3) C31-H31 0.9800
N32-C31 1.49(4) C31-C33 1.51(4)
034-H34 0.8200 N32-H32A 0.8900
The independent part of the unit cell contains two molecules of tryptophan CnH12N2O2, two molecules of water H2O, sodium cation Na+, and iodine polyanion (I4)"2. At the same time, one of the tryptophan molecules is positively charged (C11H13N2O2)+ due to the addition of a proton. The negative charge of the polyanion (I4)-2is compensated by the positive charge of tryptophanium (C11H13N2O2)+and Na+ cation (Figure 2).
In three-dimensional packing of the crystal structure of the compound C„H12N2O2KC„H13N2O2)^Na^2H20Kl4)2", Na+ cations are octahedral coordinated by six oxygen atoms, forming infinite tapes in the plane direction (Figure 3).
Figure 2 - Model of atomic structure C„H12N2O2KC„H13N2O2)^Na^2H2O^L,)2-. Ellipsoids of anisotropic thermal oscillations are shown at the 50% probability level, hydrogen atoms and disordered polyanions (I4)-2are not demonstrated.
Figure 3 - Perspective view of the crystal structure C„H12N2O2KC„H13N2O2)^NaMH2OKL,) 2-.
According to the obtained structural data, a search was conducted in the Cambridge Structural Database in order to establish the originality of the results or find analogues in the case of their availability. The search indicated that these crystals are original, were not previously obtained and studied, and new crystallographic data in the CIF format were deposited in the Cambridge Crystallographic Data Centre, the deposit number is CCDC 1877292.
The composition of the semiorganic complex compound formed in the tryptophan-sodium iodide-iodine-water system indicates that the system is non-equilibrium. The iodine hydrolysis reaction takes place in it.
H2O +I2 ^HI + HOI 2HOI ^ 2HI +O2T
(1) (2)
The resulting hydroiodic acid protonates part of the tryptophan molecules at the nitrogen atom of -NH2 -group. Therefore, in the equilibria in the tryptophan-NaI-I2-H2O system, along with the initial components there are protonated tryptophan and triiodide ions, which are formed by the reaction I2 + I- ^ I3-.
Na+ cation coordinates one tryptophan molecule and one protonated tryptophan molecule (tryptophanium) on the oxygen atoms of the carboxyl groups, as well as two water molecules.Iodide ions formed during dissociation of sodium iodide and iodide ions of hydroiodic acid form with molecular iodine tetraiodide I42-.
OH
+ 2HI
OH
+ I- (3)
The bond length in the iodine molecule (I1-I2, Table 2) is 2.762 A, while the bond length of ions I3- and I4attached to the molecule is as follows: I1-I3 -3.431 A I2-I4 - 3.628 A. In the polyanion I4- a rare case is realized when two iodide anions polarize the electrons of the iodine molecule with the formation of a partially positive charge on each atom
I- 3.431Ai8+ 2.762A 15+ 3.628 Ap-
(4)
Determination of the physical-chemical parameters of the compound. UV spectra, a 0.1% aqueous solution of tryptophan as the control and a 0.05% aqueous solution of the obtained iodine complex were measured. The UV spectrum of tryptophan displays two peaks at 218.45 nm at 0.83 A and 278.41 at 0.13A (Figure 4).
O
O
nm
Name Description - Триптофан 0,1%_250p.Sample H2O
Figure 4 - UV-spectrum of tryptophan.
The pictures below show the UV spectrum of tryptophan and tryptophan complexes: Nal: iodine. The spectra show real absorption bands that do not exist in the spectra of a pure tryptophan sample. These are bands at 190 and 280 nm for the corresponding complex formed by the reaction of tryptophan and potassium iodide with iodine (Figure 5).
l194,30nm; 1,46A|
200 300 400 500 600 700 800 900
nm
Name Description
Figure 5 - UV-spectrum of di- ((2S) -2-amino-3- (1H-indol-3-yl) propionate)-dihydro-tetraiodide.
The thermograms were recorded differential curves of change in the heat capacity of the sample. The DSC curves of tryptophan shows the peak of melting point of tryptophan is 298.1°C. Starting value of the melting point at the peak is 292°C. The table data for the melting point of tryptophan is 293-295°C (Figure 6).
Figure 6 - DSC and TG curves of Tryptophan.
The DSC and TG curves of complex shows the peak of melting point of complex is 58.9°C (Figure 7). The TG curve of the new complex shows us the single stage decomposition.
TG /% DSC /(jiVimg)
Figure 7 - DSC and TG curves of di- ((2S) -2-amino-3- (1H-indol-3-yl) propionate)-dihydro-tetraiodide.
Some physicochemical parameters of the di-((2S)-2-amino-3- (1H-indol-3-yl) propionate)dihydro-tetraiodide complex is presented at Table 3.
Table 3 - Physicochemical parameters of the di- ((2S) -2-amino-3- (1H-indol-3-yl)propionate)-dihydrotetraiodide complex
Parameter Value
Particleshape elongated needle-like linear stick-like shape
Bulkdensity, g/cm3 0.767
Particlesize, ^m 3.20
Solubility It is soluble in water, DMSO, slightly soluble in ethanol, acetone and cyclohexane.
pH of aqueous solutions 4.30-4.40
Solubility studies show that the complex dissolves well in solvents with a polarity index of 7.2 and 10.2, and in solvents with an index of 5.1; 4.3 and 0.04 are slightly soluble. The dissolution process in organic solvents is accompanied by the formation of an intense brown color.
Cytotoxicity of the complex. The cytotoxic effect of the coordination compound was studied to determine the maximum concentrations that do not have toxic properties. Monolayer transplantable cell culture MDCK was used to evaluate the toxicity. The results of the study assessing the cytotoxic effect of the coordination compound on MDCK cell culture are presented in Figure 8.
0.0156 0.0313 0.0625 0.125 0.250 0.500 1.000 2.000
Concentration, mg/m)
Figure 8 - Cytotoxic effect of the coordination compound on MDCK culture.
The toxicity value for the coordination compound was 1.70 mg/mL, which characterizes the examined substance as a low toxic compound.
Antimicrobial activity of complex. The results of testing for antimicrobial activity are presented in Table 4.
As may be seen from the above, the semiorganic complex contains molecular iodine in its structure, which has antimicrobial properties. Its content in the complex is 26 mass%, and the coordination compound is therefore of particular
Table 4 - Minimum bactericidal concentrations values of the di-((2S -2-amino-3-(1H-indol-3-yl) propionate)-dihydro-tetraiodide, ^g/mL
Sample Test strain Minimum bactericidal concentrations of the complex, ^g/mL
C„H12N2O^ (C11H13N2Ü2)+ • Na+^2H20 •(L) 2" S. aureus ATCC 6538-P 125
S. aureus ATCC-BAA-33591 125
E. coli ATCC 8739 250
E. coli ATCC-BAA 2523 125
P. aeruginosa ATCC 9027 250
P. aeruginosa TA 2 250
interest due to its potential antimicrobial activity. This compound is effective against both susceptible and resistant strains of S. aureus ATCC 6538-P and S. aureus ATCC-BAA-33591, value of the minimum bactericidal concentration is125 ^g/mL. The coordination compound is effective against the susceptible P. aeruginosa strain ATCC 9027, as well as the clinical multiresistant of P. aeruginosa strain TA2, at a concentration of 250 ^g/mL.
The minimum bactericidal concentrations of the compound under testing against the susceptible E. coli strain ATCC 8739 and multiresistant E. coli strain ATCC-BAA 2523 are 250 ^g/mL and 125 ^g/mL, respectively. The complex exhibits bactericidal activity against both susceptible and multiresistant bacterial strains.
Determination of the mutagenic activity of the complex in the in vitro micronucleus assay. In these studies, the maximum concentration used is 5 mg / ml, followed by 2-fold dilution in exposure medium. The analysis is carried out with a 4-hour exposure with metabolic activation (+ S9) and without metabolic activation (- S9). For the analysis of micronuclei, slides with L5178Y cells treated with complex di-((2S -2-amino-3- (1H-indol-3-yl)propionate)-dihydro-tetraiodide at concentrations from 0.625 to 0.039 mg/ml in the presence or absence of metabolic activation are taken.
Calculation of the number of micronuclei per 1000 mononuclear cells with new complex shows (Figure 9) an unreliable increase in the frequency of micronuclei with an increase in the concentration of complex in the presence and absence of metabolic activation.
Evaluation of the results of the experiment. If the validity of the analysis is satisfied, the following criteria are met:
- Positive response: a test substance is considered a mutagenic substance if it induces a statistically significant, dose-dependent increase in the frequency of mononuclear cells compared to a negative control.
- Negative response: a test substance is considered a non-mutagenic substance if there is no statistically significant increase in the frequency of mononuclear cells compared to a negative control.
Figure 9 - Frequency of micronuclei in the presence of a new complex.
Thus, the new complex does not cause any mutagenic effect on mammalian cells of the L5178Y line, both in the presence and in the absence of metabolic activation.
4. Conclusion
New iodine complex from system tryptophan:NaI:iodine:water system was synthesized. The parameters of triclinic unit cell were determined. As follows from the obtained data, a new complex di- ((2S) -2-amino-3- (1H-indol-3-yl) propionate)-dihydro-tetraiodideor C11H12N202^(C11H13N202)+^Na+^2H20^(I4) 2-has low cytotoxicity, direct antiviral effect, antimicrobial activity against antibiotic-resistant strains of microorganisms and is therefore promising for further use as an active antimicrobial substance. New complex does not cause any mutagenic effect on mammalian cells of the L5178Y line, both in the presence and in the absence of metabolic activation.
Acknowledgments. The research was carried out with the financial support of the Republic of Kazakhstan for a fundamental research on the topic "Development of new anti-infectious drugs".
The authors express their gratitude to the staff of the laboratories of virology and microbiology of JSC «Scientific center for anti-infectious drugs», Almaty, Republic of Kazakhstan who provided assistance in preparing the materials of the article.
Information about authors:
Sabitov A.N. - Cand. of chemical sciences (PhD); e-mail: aitugans@mail.ru; ORCID ID: https://orcid.org/0000-0003-3677-86856
Turganbay S. - Doctor PhD; e-mail: serik530@mail.ru; ORCID ID: https://orcid.org/0000-0001-9621-3534
Jumagaziyeva A.B. - PhD Student;e-mail: r_dawa@mail.ru; ORCID ID: https://orcid.org/0000-0002-8610-7321
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Тушндеме
ДИ-(^)-2-АМИН-3-(1Н-ИНДОЛ-3-ИЛ)ПРОПИОНАТ)-ДИГИДРОТЕТРАИОДИДТЩ ЦУРЫЛЫМЫ ЖЭНЕ ЦАСИЕТТЕР1
А.Н. Сабитов, С. Турганбай, A. Джумагазиева
Тестшеу зауыты, АК «Инфекцияга царсы дэр^дэрмектер орталыгы», Алматы, Казацстан
E-mail: aitugans@mail.ru
Амин кышкылы - сштшк метал тры - иод - су жуйей негiзiнде жаца органи-кальщ комплекс синтезделд^ ди-((28)-2-амино-3-(1Н-индол-3-ил) пропионат) -дигидро-тетраиодид ретшде аныкталган комплекстщ физика-химиялык касиеттерi зерттелдi. Микроскопиялык талдау керсеткендей, комплекстiн бвлшектерi :рын ине тэрiздi сызыкты таякша тэрiздi пiшiнде, орташа мвлшерi 2.50-4.00 мм. Комплекстщ микробка карсы белсендiлiгi S. aureus ATCC 6538-Р (м^ражайсезiмтал штаммы); S. aureus ATCC-BAA-39 (м^ражайдыц теракты штаммы); E. coli ATCC 8739 (м^ра-жай сезiмтал штаммы); E. coli ATCC-BAA-196 (м^ражайдыц теракты штаммы); P. aeruginosa ATCC 9027 (м^ражай сезiмтал штаммы); P. aeruginosa TA2 жасушала-рында тiкелей вируска жэне антибиотикке твзiмдi микроорганизмдердiн штамда-рына карсы белсендiлiгi бар екенi аныкталды. MDCK жасушасына жасалган зерттеу нэтижесшде цитотоксикалык эсерiнiн твмендiгi кврсетiлдi. Di-((2S)-2-амино-3-(1H-индол-3-ил)пропионат)-дигидротетраиодид L5178Y сызыкты CYткоректiлер жасу-шаларына катысты мутагендiэсер етпейдi.
Тушн свздер: иод полианион кешенi, ди-((28)-2-амин-3-(1Н-индол-3-ыл) про-пионат)-дигидротетраиодид, рентгендiк к¥рылымдык талдау, L5178Y сызыкты жа-сушаларына мутагендж тест, цитотоксикалык эсер.
Резюме
СТРУКТУРА И СВОЙСТВА ДИ-(^)-2-АМИН-3-(1Н-ИНДОЛ-3-ИЛ) ПРОПИОНАТ)-ДИГИДРОТЕТРАИОДИДА
А.Н. Сабитов, С. Турганбай, A. Джумагазиева
Испытательная база АО «Научный центр противоинфекционных препаратов», Алматы, Казахстан E-mail: aitugans@mail.ru
Синтезирован новый комплекс йода в системе аминокислота - соль щелочного металла - йод - вода. Изучены физико-химические свойства комплекса ди-((28)-2-амино-3 -(1Н-индол-3 -ил)пропионат)дигидротетраиодид. Микроскопический анализ показывает, что частицы комплекса имеют продолговатую игольчатую линейную
палочковидную форму со средним размером 2,50-4,00 мкм. Определяли цитоток-сичность на культуре клеток MDCK, антимикробную активность на S. aureus ATCC 6538-Р (музейно-чувствительный штамм); S. aureus ATCC-BAA-39 (музейный мультирезистентный штамм); E. coli ATCC 8739 (музейно-чувствительный штамм); E. coli ATCC-BAA-196 (музейный мультирезистентный штамм); P. aeruginosa ATCC 9027 (музейно-чувствительный штамм); P. aeruginosa TA2. Комплекс обладает низкой цитотоксичностью, прямым противовирусным дейст-вием и антимикробной активностью в отношении устойчивых к антибиотикам штаммов микроорганизмов. Ди-((2S)-2-амино-3-(1H-индол-3-ил)пропионат)дигидротeтраиодид не оказывает мутагенного действия на клетки млекопитающих линии L5178Y как в присутствии, так и в отсутствии метаболической активации.
Ключевые слова: иод-полианионный комплекс, ди-((2S)-2-амино-3-(1H-индол-3-ил)пропионат)дигидротетраиодид, рентгеноструктурный анализ, мутагенный тест на клетках L5178Y, цитотоксичность.
