Thus, researches of recent years point out a special attention towards therapeutic combinative effect of pyridoxine and zinc. During the development and introduction of new medications into medical practice, pharmacological researches play a special part, as their results allow one to receive accurate quantitative data on absorption, distribution, metabolism, and extermination of a medicine, and also show a correlation dependence of pharmacologic activity and pharmacologic parameters of a medication [1, 2].
Pyracin is a complex solution of zinc microelement with pyroxidin. Due to pharmacologically rational doze of the preparation components, Pyracin has a wide range of therapeutic effect. Studies of metabolic activity of the preparation show that Pyracin has hypolipidemic and anti-atherosclerotic activity, decreases contents of cholesterol in periphery blood, general lipids, treglycerides, lipoproteins of low density, products of peroxide oxidation of lipids, increases contents of lipoproteins of high density and phospholipids. Under sugar diabetes with hyperlipidemia, the preparation decreases contents of glucose, pyruvate, and lactic acid in blood. Besides, the preparation is widely used in treating dermatologic-venereologic pathologies in terms of skin displays that are linked to a deficit of zinc (vitiligo, inflammatory and seborrheic alopecia, pustular and phegmonous acne, acrodermatitis enteropathica, difficult-to-heal skin ulcers) [3].
Objective: pharmacologic kinetic research of acute toxicity of the medical preparation «Pyracin-RG», injection solution 0,25 %, produced by Joint Enterprise «Remedy Group», Tashkent, Republic Uzbekistan.
Finding a preparation dose for pharmacologic kinetic research was carried out via studying an acute toxicity of «Pyracin-RG». The research has been implemented on white pedigreeless mice of both sexes of mass 18–20 g, 6 animals per a group, total of 24 mice have been used.
The tested solution was introduced to animals one time hypodermically in dose: 15; 2; 22,5; 25 ml/kg.
After a single introduction of the medication, the animals were observed hourly on the day of introduction, 3 times a day on days 2–3 after the introduction, and once a day during the following 7 days of the research. We have examined general condition, behavior, fur pigment, breath, heartbeat, moving activity, and death of the mice.
Under the dose of 15 ml/kg, the first 15 minutes we have been observing breath of increased frequency, grouping, the mice moved slowly, scratched and washed. Under the dose of 20 ml/kg, during the first 45 minutes, a heavy short breath, slow movement, itching, and grouping has been observed. The dose of 22,5 ml/kg led to a heavy short breath, immobility, grouping during the first 1,5 hours, and also strong itching during the first 15 minutes.
Under the dose of 25 ml/kg a strong itching, heavy short breath, immobility, grouping has been observed during the first 2 hours.
Death of animals has not been observed after the introduction of «Pyracin-RG» (Table 1).
Therefore, the preparation does not have a lethal-toxic effect in the studied doses.
Pharmacologic-kinetic parameters of the medication have been estimated according to a quantitative content of zinc in organs and tissues.
43 pedigreeless white male rats of body mass of 180–200 gr have been used in the experiment. A control group has been formed of 3 animals, definition of pyracin in fecal mass has been carried out among 5 animals, and 5 rats have been selected at each checkpoint. The studied preparation was introduces hypodermically to the experiment rats in dose of 5 ml/kg. Tests of blood and organs have been received in 30 minutes, 1, 2, 3, 4, 6, and 24 hours after decapitation of animals. Fecal masses were also collected once available, as the main criterion of zinc excretion is gastro-intestinal tract. Fecal losses of zinc consist of non-absorbed and secreted zinc endogen.
Table 1
Results of defining an acute toxicity of «Pyracin-RG»
|
Number |
Dose (ml/kg) |
Number of animals/total dead (units) |
|
1. |
15 |
6/0 |
|
2. |
20 |
6/0 |
|
3. |
22,5 |
6/0 |
|
4. |
25 |
6/0 |
|
Lethal Dose50 > 25 ml/kg |
||
Organs, blood serum, and faeces have been analyzed via method of optical-emission spectrometry with inductively-linked argon plasma. 0,500–2,000 of the examined organ have been weighed and transported to teflon autoclaves. 3 ml of nitrous acid and 2 ml of hydrogen peroxide have been added to autoclaves, then they were closed and placed into a facility of microwave dissolution BERGHOF, equipped with the software Speebwaveтм MWS-З4+. After the dissolution contents of autoclaves were transited into a 50 ml measure retort and have been added up to the mark of 2 % nitrous acid.
Definition of the studied substance has been carried out on optical-emission spectrometer with inductively linked argon plasma Optima-2100 DV (USA). Optimal wave length of the defined channel Zn has been used, with it the element discharges its maximum emission of the absorbed energy. Quantitative content of the studied element Zn in samples was calculated according to the formula (mkg/g):
Zn = N∙100/C,
where N is a quantity of metal from the graphic (mg/kg); C is a concentration of the studied sample in the prepared solution.
Organs and faeces of rats contain certain amount of zinc. Therefore, via excluding control data from the experiment rats, we find the quantity of zinc that has penetrated them with the preparation (mkg/g).Considering zinc contents in pyracin, we have re-calculated the preparation dose and established its concentration on the studied object. Multiplying the received results by weight of each organ, we have calculated contents of pyracin in an organ, it is shown in Table 2.
Table 2
Quantitative contents of Pyracin in organs in faeces of rats (mkg/organ)
|
Number |
Checkpoints (minutes) |
||||||
|
30 minutes |
1 hour |
2 hours |
3 hours |
4 hours |
6 hours |
24 hours |
|
|
Liver |
|||||||
|
1 |
705,1 |
1095 |
30,779 |
257,28 |
716,38 |
756,74 |
282,67 |
|
2 |
607,4 |
1072 |
29,74 |
287,72 |
727,79 |
703,6 |
343,57 |
|
3 |
584,5 |
932,6 |
24,577 |
346,81 |
843,02 |
828,43 |
288,32 |
|
4 |
570,2 |
959,4 |
24,725 |
347,1 |
842,18 |
726,1 |
308,74 |
|
5 |
537,7 |
842,9 |
21,383 |
340,28 |
781,25 |
797,28 |
298,47 |
|
Average |
601,0 ± 63,47 |
980,40 ± 103,9 |
26,241 ± 3,92 |
315,84 ± 41,089 |
782,12 ± 60,398 |
762,43 ± 50,95 |
304,35 ± 24,076 |
|
Kidney |
|||||||
|
1 |
52,01 |
56,24 |
54,758 |
45,636 |
83,135 |
59,157 |
19,509 |
|
2 |
43,29 |
40,64 |
52,353 |
60,617 |
94,111 |
44,23 |
25,013 |
|
3 |
38,4 |
44,46 |
57,745 |
73,903 |
125,42 |
50,717 |
21,073 |
|
4 |
34,3 |
43,85 |
48,650 |
72,144 |
107,49 |
54,716 |
24,555 |
|
5 |
36,72 |
56,12 |
44,479 |
67,145 |
113,83 |
48,852 |
24,537 |
|
Average |
40,940 ± 7,007 |
48,260 ± 7,372 |
51,5976 ± 5,188 |
63,889 ± 11,433 |
104,800 ± 16,565 |
51,534 ± 5,68 |
22,937 ± 2,48 |
|
Lungs |
|||||||
|
1 |
91,82 |
15,16 |
1,327 |
– |
– |
– |
– |
|
2 |
121,1 |
11,34 |
1,358 |
– |
– |
– |
– |
|
3 |
79,09 |
8,832 |
0,811 |
– |
– |
– |
– |
|
4 |
91,52 |
11,53 |
1,132 |
– |
– |
– |
– |
|
5 |
110 |
10,3 |
1,225 |
– |
– |
– |
– |
|
Average |
98,71 ± 16,69 |
11,43 ± 2,34 |
1,171 ± 0,21 |
– |
– |
– |
– |
|
Lien |
|||||||
|
1 |
2,893 |
7,259 |
128,835 |
22,963 |
43,625 |
20,184 |
7,6677 |
|
2 |
2,252 |
8,845 |
109,591 |
27,706 |
43,754 |
32,539 |
17,309 |
|
3 |
4,046 |
10,39 |
132,89 |
30,136 |
38,11 |
40,172 |
12,327 |
|
4 |
3,466 |
7,617 |
97,7924 |
41,746 |
38,809 |
31,505 |
14,075 |
|
5 |
2,757 |
4,658 |
81,99 |
48,852 |
46,979 |
29,585 |
13,111 |
|
Average |
3,083 ± 0,69 |
7,754 ± 2,12 |
110,22 ± 21,28 |
34,28 ± 10,683 |
42,255 ± 3,72 |
30,797 ± 7,16 |
12,898 ± 3,48 |
|
Faeces |
|||||||
|
1 |
1,291 |
– |
– |
– |
– |
125,674 |
1830,598 |
|
2 |
– |
– |
12,573 |
– |
– |
123,707 |
1924,699 |
|
3 |
0,627 |
– |
– |
– |
– |
130,456 |
1794,714 |
|
4 |
1,047 |
– |
14,387 |
– |
– |
127,464 |
1728,33 |
|
5 |
– |
– |
8,997 |
– |
– |
128,44 |
1742,372 |
|
Average |
0,988 ± 0,335 |
– |
11,985 ± 2,742 |
– |
– |
127,14 ± 2,58 |
1804,143 ± 78,87 |
|
Blood (mkg/ml) |
|||||||
|
1 |
10,794 |
18,966 |
20,406 |
73,628 |
24,062 |
3,934 |
– |
|
2 |
10,730 |
19,396 |
20,600 |
73,134 |
24,040 |
3,848 |
– |
|
3 |
10,772 |
19,246 |
20,492 |
73,522 |
24,084 |
4,342 |
– |
|
4 |
10,622 |
19,482 |
20,126 |
73,780 |
24,126 |
4,300 |
– |
|
5 |
10,686 |
19,352 |
20,642 |
73,306 |
24,040 |
4,256 |
– |
|
Average |
10,72 ± 0,068 |
19,29 ± 0,18 |
20,45 ± 0,20 |
73,46 ± 0,26 |
24,06 ± 0,02 |
4,136 ± 0,22 |
– |
According to the received data on the amount of pyracin in rat blood we have carried out calculations of pharmacological-kinetic parameters for the studied preparation in the application Borgia [4, 5]:
Тmax – period of achieving maximum concentration, minutes;
Cmax – maximum concentration, mkg/ml;
Period of half-absorption, K ½, minutes;
Clearance, Cl, ml/min;
Area under the curve, AUC;
Average period of hold, MRT (0-∞).
Pharmacological parameters are provided in Table 3.
Table 3
Pharmacological-kinetic parameters for «Pyracin-RG», solution of 0,25 %
|
Pharmacological-kinetic parameters |
Parameter indexes for serum |
|
Тmax, minutes |
162,4 ± 0,894 |
|
Сmax, mkg/ml |
75,123 ± 0,375 |
|
Ka1/2, minutes |
53,801 ± 1,041 |
|
Т1/2, minutes |
41,992 ± 0,230 |
|
Cl, ml/min |
0,0438 ± 0,0004 |
|
AUC |
22529 ± 732,1 |
|
MRT (0-∞) |
19,54 ± 0,151 |
Thus, the received research data of the amount of pyracin in organs and faeces of rats in definite time periods – 30 minutes, 1, 2, 3, 4, 6, and 24 hours after the introduction of medicine in calculations (mkg/g) show its concentration in blood, liver, kidney, lungs tends to decrease in later periods of the experiment in comparison to its earlier periods. On the other hand, concentration of pyracin in lien and faeces tends to increase, obviously, due to the maximum excretion and accumulation of zinc solutions in the studied biomaterials.
The provided research has shown that in 24 hours after the medicine introduction, 72 % of pyracin is removed from rat organism with faeces. In 24 hours after the introduction the medicine cannot be found in an organism. Maximum contents of pyracin in lungs are observed is observed during the first 30 minutes after the introduction, and it is removed in 3 hours. Maximum concentration of pyracin in lien is observed in 2 hours after the introduction, and in 24 hours only 0,05 % of the preparation remains. Maximum content of pyracin in kidney is observed in 4 hours after the introduction. After 24 hours of observation 0,9 % of the preparation remains in kidney. Concentration of pyracin in liver reaches its maximum concentration in 4-6 hours after the introduction, and 1,23 % of the medicine remains in it after 24 hours.
Resume
1. «Pyracin-RG» 0,25 % solution does not lead to death of an animal after a single introduciton.
2. Results of the pharmacological-kinetic research show that a maximum concentration of pyracin in blood serum equals 75, 1,23 mkg/ml, period of achieving it – 2 hours 40 minutes, and period of its half-distribution in blood equals 42 minutes. In 24 hours after the introduction Pyracin cannot be found in blood. 72 % of the preparation is removed from rat organism with faeces.
3. Study of Pyracin distribution dynamics throughout organs has shown that maximum contents of pyracin in lungs is observed is observed during the first 30 minutes after the introduction, and it is removed in 3 hours. Maximum concentration of pyracin in lien is observed in 2 hours after the introduction, and in 24 hours only 0,05 % of the preparation remains. Maximum content of pyracin in kidney is observed in 4 hours after the introduction. After 24 hours of observation 0,9 % of the preparation remains in kidney. Concentration of pyracin in liver reaches its maximum concentration in 4–6 hours after the introduction, and 1,23 % of the medicine remains in it after 24 hours.