RU2187309C2 - Agent effecting on arterial pressure - Google Patents

Abstract

FIELD: organic chemistry, medicine, pharmacy. SUBSTANCE: invention claims an agent comprising S-alkylisothiurnium as an active substance of the formula

, where Alk is the branched or nonbranched C₁-C₄-alkyl and x^- is anion of phosphorus- containing acid. A new agent is deprived of undesirable adverse effects typical for adrenomimetics exhibiting the same action. Invention can be used for production of preparations that effect on arterial pressure. EFFECT: expanded assortment of agents with hypertensive effect, valuable medicinal property. 3 cl, 14 dwg, 13 tbl, 3 ex

Description

 The invention relates to the field of meeting the vital needs of a person, namely to new drugs that can be used in the pharmaceutical, pharmaceutical and medical industries, specifically to phosphorus derivatives of isothiuronium, including new chemical compounds that affect blood pressure, which have a vasoconstrictor used in cases of chronic and acute hypotension (bleeding, trauma, shock, poisoning), including especially in emergency and emergency care bitch.

 In medical practice, medicinal substances of pressor action are used, which act either due to the stimulation of alpha-adrenergic receptors, or due to the direct effect on the smooth muscles of the vascular wall. Most often, adrenomimetics are used, for example, adrenaline, norepinephrine, mesatone, methoxaline, ephedrine, fetanol, etc., and polypeptide preparations, for example, glucagon, antioteninamide, octapressin, etc.

 According to pharmacological properties, the closest analogue to the proposed compound is the well-known drug mesatone (M.D. Mashkovsky. Medicines, ed. 12th, M., Medicine, 1993, part I, p. 303).

Mesatone is 1- (m-hydroxyphenyl) -2-methylaminoethanol hydrochloride. It selectively stimulates α ₁ -adrenoreceptors, causes narrowing of arterioles and an increase in systolic and diastolic pressure (with possible reflex bradycardia). Mesatone has virtually no pacemaking effect. Unlike adrenaline and norepinephrine, mesatone is not a catecholamine (it contains only one hydroxyl group in the aromatic nucleus) and is little susceptible to the action of the enzyme catechol-O-methyltransferase, and therefore more resistant, has a longer effect. The antihypertensive effect of mesatone usually lasts about 20 minutes after a single intravenous administration (M.D. Mashkovsky, Medicines, ed. 12th, M., Medicine, 1993, part I, p. 303; O. M. Avakyan, Pharmacological regulation function of adrenergic receptors, M., Medicine, 1988, p. 8).

 Adrenergic agonists, including mesatone, have some common drawbacks, as they increase oxygen consumption by tissues, cause metabolic acidosis, can cause arrhythmias (especially during general anesthesia), have an exciting effect on the central nervous system (O.M. Avakyan, Pharmacological regulation of adrenoreceptor function , M., Medicine, 1988, p. 8, V.G. Kulinsky, A.N. Kovalevsky, Bul.experim.biol.med., 1984, p. 9, “Dualistic control of oxygen consumption by catecholamines in the body of mice”). They are characterized by the occurrence of secondary hypotension. Adrenergic agonists do not correct arterial hypotension caused by adrenergic blockers, under conditions of metabolic acidosis, they have only a weak antihypertensive effect (C. Kortanje, VIMathy, R. Chartdorp, Haunyn-Gohinedeleg is Arh. Pharmacol., V.330, N3, p. 187-192 (1985), "Influence respiratory acidosis or alkalosis on pressor responses imediated by and alfa-adrenoreceptors in pithed normotensive rats", IMAutkunson, SIDusting, VIRand, Aust.J.Exp.Biol.Med., V.50, p .847-859 (1972), "Acidosis induced by catecholamines in acidosis").

 Preparations with a polypeptide structure and most adrenomimetics have a short duration of action and are administered in the form of perfusion to achieve a prolonged effect (IMAutkunson, SIDusting, VIRand, Aust. J.Exp.Biol.Med., V.50, p.847- 859 (1972), "Acidosis induced by catecholamines in acidosis").

 The chemical synthesis of phosphorus-containing derivatives of S-alkylisothiuronium was first described by the interaction of thiourea with trialkylphosphates or dialkylphosphites when heated in an organic solvent, however, the structure of N-alkylthiuronone phosphates was attributed to the obtained compounds (JBParker, T. D. Smith, J. Chem. Soс., 1961, p. .442-445, "The reaction of thiourea with dialkyl phosphites and trialkyl phosphates"). Later, their structure was correctly established as phosphate or phosphinates of S-alkylisothiuronium) (P.G. Zherebchenko, Yu. D. Zilber, G.P. Posepekhova et al., Radiobiology, vol. 8, pp. 582-587 (1968) , "Radioprotective effect of salts of S-alkyl substituted isothiourea derivatives in their separate and combined study") and was confirmed by physicochemical methods of analysis (L.I. Mizrakh, V.G. Yakovlev, E.M. Yukhno et al., MLC, t .41, pp. 2654-2658 (1971), “Some phosphorus-containing derivatives of isothiuronium”), but a detailed method for their preparation has not yet been described.

 The literature describes that phosphorus-containing derivatives of S-alkyl isothiuronium have a pronounced radioprotective effect (P.G. Zherebchenko, Yu.D. Zilber, G.P. Posepekhova and others. Radiobiology, vol. 8, pp. 582-587 (1968) , "Radioprotective effect of salts of S-alkyl-substituted isothiourea derivatives in their separate and combined study"; N.A. Izmozherov, Yu.V. Gilev, L.I. Mizrakh and others. Abstracts at the 2nd All-Union Conference on Pharmacology of Radiation preparations, M., 1972, pp. 129-133, "Modification of radiation death of animals using phosphorus-containing derivatives isothiuronium alone "; N.A. Izmozherov, L.I. Mizrakh, V.V. Kozhevnikov et al., ibid., pp. 133-136," Radiolucency at the cellular level of phosphorus-containing isothiuronium derivatives "; T.Yu. Ilyuchenok , L.M. Frigidova, Yu.V. Zavyalov et al. Pharmacology and Toxicology, vol. 39, pp. 191-198 (1972), "Radioprotective properties of some phosphorus-containing derivatives of isothiuronium"; Zh. A. Goloshchapova, Radiation exposure , recovery and chemical protection. Sverdlovsk, 1978, pp. 71-75, "Toxic and radioprotective properties of certain phosphorus-containing derivatives of isothiuronium"; Zh.A. Goloshchapova, T.N. Tuzhilkova, L.I. Mizrakh, Radiobiology, t.21, p. 521-525 (1981), "Experimental justification for the use of some derivatives of isothiuronium as radioprotective compounds") and some toxicological specifications.

 The objective of the invention is the search for substances, including new compounds with hypertensive activity and devoid of these disadvantages of adrenergic agonists.

где Alk представляет собой разветвленный или неразветвленный алкил, а Х^- - анион фосфорсодержащей кислоты, в качестве средств, влияющих на артериальное давление.The problem is solved using phosphorus-containing derivatives of S-alkylisothiuronium, including new ones not described in the literature, of the general formula

where Alk is a branched or unbranched alkyl, and X ^is an anion of phosphorus-containing acid, as agents that affect blood pressure.

 Their synthesis is carried out by known methods, most often by alkylation of thiourea with the corresponding phosphorus derivatives, the salt exchange reaction was rarely used.

 As can be seen from the data on biological studies below (Fig. 1-10, Table 1) of phosphorus-containing derivatives of S-alkyl isothiuronium, the latter have a pronounced, as a rule, quick and lasting effect on blood pressure.

Of particular note are the new chemical compounds:
S-methylisothiuronium dimethyl phosphate,
S-isopropylisothiuronium metaphosphate,
isobutylphosphite S-isobutylisothiuronium.

 It is useful to emphasize that S-isopropylisothiuronium metaphosphate, significantly increasing blood pressure, does not affect the frequency of respiratory movements.

 The diethylphosphate of S-ethylisothiuronium, the effect of which is presented in Table 2, has been studied in most detail. 2-5 and FIG. 11-14.

 The duration of the action of diethylphosphate S-ethylisothiuronium in comparison with adrenomimetics (for example, mesatone) increases by 2-3 times (Table 2-5, Fig. 11-13). The proposed compound does not negatively affect the acid-base balance (Tables 6, 7), does not cause secondary hypotension (Figs. 11-13), has specific activity under the action of α-blockers (Table 3), practically does not reduce the effectiveness of metabolic acidosis (table. 4), improves the performance of systemic hemodynamics in hemorrhagic shock (table. 4).

 Under the action of diethylphosphate S-ethylisothiuronium, the indicators of systemic hemodynamics change as follows: peripheral vascular resistance increases, the stroke volume and central volume of the circulating blood increase, the work of the left ventricle increases (Table 4).

 In case of shock conditions (traumatic, hemorrhagic), intramuscular administration of diethylphosphate S-ethylisothiuronium allows you to provide assistance at the prehospital stage (Table 4), which cannot be achieved with the currently used drugs.

 It is recommended to use diethylphosphate S-ethyl isothiuronium as a drug to increase blood pressure in acute arterial hypotension caused by surgical interventions, injuries, poisoning, shock conditions, bleeding, with epidural anesthesia, with an overdose of ganglion blockers (Table 5), alpha-blockers (Table 5), alpha-blockers (Table 5), . 3), antipsychotics, anesthetics, and in other conditions when adrenergic agonists are contraindicated or ineffective (Tables 6, 7).

 Interestingly, the effect of diethylphosphate S-ethylisothiuronium is stronger in the case of animal hypotonics than for normotonics (Figs. 12, 13).

 All these data show that the proposed compounds differ from known agents that are used to increase blood pressure.

 Currently, documentation is being prepared for diethylphosphate S-ethylisothiuronium for submission to the Pharmacological Committee of the Ministry of Health of the Russian Federation for permission to conduct its clinical trials called Difetur, the rest of the substances undergo extensive biological and toxicological studies.

 The use of the proposed compounds in medical practice creates new opportunities in the treatment of acute and possibly chronic arterial hypotension, as well as in cases of emergency and emergency care.

 Phosphorus-containing derivatives of S-alkylisothiuronium can be used in a dosage form convenient for a particular case. Most often, it is proposed to use drugs in the form of tablets, ampoules for intramuscular or intravenous administration, however, other dosage forms, such as suppositories, can be used. In emergency conditions, it is recommended to administer drugs intravenously or intramuscularly in the form of 1-10% aqueous solutions at a dose of 0.1-20 mg / kg body weight, usually 0.2-5 mg / kg.

Phosphorus derivatives of S-alkylisothiuronium are low toxic (LD ₅₀ is 100-1000 mg / kg).

 Relative contraindications are: arterial hypertension, pheochromocytoma, pregnancy. With caution, you need to prescribe the drug to the elderly.

 In case of drug overdose, intravenous administration of calcium antagonists (verapramil) or papaverine is recommended.

 The following experimental data illustrate but do not limit the claims of the applicant.

 Synthesis of phosphorus derivatives of S-alkylisothiuronium.

 Diethyl phosphate S-ethylisothiuronium.

36.6 g (0.201 M) of triethyl phosphate and 15.2 g (0.2 M) of pre-ground thiocarbamide are charged into the flask. The reaction mass with stirring is heated to 132-135 ^o C and maintained at this temperature until complete dissolution of thiocarbamide (about 3-5 minutes). Continuing stirring, heating is stopped and the mixture is allowed to cool spontaneously. When the reaction mixture reaches a temperature of 50-60 ^o With 40 ml of acetone is poured to it. With a further decrease in temperature, crystallization of the product begins, which ends with stirring at room temperature in 3 hours. The crystals are filtered off, washed with 10 ml of acetone and dried to constant weight. 26.6 g of crude S-ethylisothiuronium diethyl phosphate are recovered, which are recrystallized from acetone. Obtain 24.6 g (47.7% of theory) of diethylphosphate S-ethylisothiuronium with so pl. 144-146 ^o C, literature data so pl. 145 ^o C (JB Parker, 1961), the content of the main substance of 99.6%.

Found,%: C 32.57, H 7.43, N 10.75, P 11.87, S 12.47. C ₇ H ₁₉ N ₂ O ₄ PS. Calculated,%: С 32.56, Н 7.36, N 10.85, Р 12.01, S 12.40.

Thin layer chromatography - silofol plate system:
aqueous ammonia: chloroform: propan-2-ol (1: 8: 4) R _f = 0.19;
aqueous ammonia: chloroform: propan-2-ol (3: 1: 2) R _f = 0.44.

 Isopropylphosphite S-isopropylisothiuronium.

A mixture of 13.7 g (0.082 M) of diisopropopylphosphite and 6.2 g (0.082 M) of thiourea is heated in a three-necked flask equipped with a stirrer and reflux condenser with a calcium chloride tube to a temperature of 135 ^o C. At the same time, thiourea dissolves and a homogeneous mass forms. Heating at this temperature is continued for another 60-75 minutes until the reaction mass is slightly reddened. Then the mixture is cooled to 40-50 ^o C and with a working stirrer pour 50 ml of acetone. After 30 minutes, the crude is filtered off and 10 g (50.8%) are obtained. After recrystallization from a mixture of acetone - ethyl alcohol (2: 1), 6.0 g of S-isopropylisothiuronium isopropylphosphite are obtained with a melting point of 169-171 ^o With, literature data so pl. 164 ^o C (V. V. Orlovsky, B. A. Vovsi, Zhokh, vol. 39, pp. 1259-1260 (1969). On the question of the interaction of dialkyl phosphites with thiourea).

 Metaphosphate S-ethylisothiuronium.

A mixture of 8.65 g (0.05 M) of diethyl chlorophosphate and 3.8 g (0.05 M) of well-ground thiourea is heated with stirring, raising the temperature to 145 ^o C for 1.5 hours, kept at this temperature for 10 minutes, cool, pour a mixture of ethanol - acetone (3: 2) and leave overnight. The precipitate is triturated in this solvent mixture and filtered; after crystallization from aqueous alcohol, 6.0 g (65%) of S-ethylisothiuronium metaphosphate are obtained with mp. 224-225 ^o C.

Found,%: C 19.57, H 4.90, P 16.73, S 17.32. C ₇ H ₁₉ N ₂ O ₄ PS. Calculated,%: С 19.55, Н 4.90, Р 16.85, S 17.40.

The same substance is obtained in another way:
A mixture of 4.25 g of ethyl metaphosphate and 3.0 g of thiourea with stirring is heated for 1.5 hours to 145 ^o C, maintained at this temperature for 10 minutes, cooled and treated as described above. Obtain 5.0 g (69%) of S-ethylisothiuronium metaphosphate with so pl. 223-224.5 ^o C.

Other compounds are obtained similarly:
methylphosphite S-methylisothiuronium with a yield of 73%, mp 119-120 ^o With, literature data so pl. 120 ^o C (V.V. Orlovsky, 1969),
S-methylisothiuronium dimethyl phosphate in 54% yield,
ethylphosphite S-ethylisothiuronium with a yield of 62%, so pl. 109-110 ^o With, literature data so pl. 112 ^o C (V.V. Orlovsky, 1969),
propylphosphite S-propylisothiuronium with a yield of 40%, so pl. 99-100 ^o With, literature data so pl. 102 ^o C (V.V. Orlovsky, 1969),
S-isopropylisothiuronium metaphosphate with a yield of 21%, mp 257-259 ^o C,
S-butylisothiuronium dibutyl phosphate in 38% yield, mp 96-98 ^o With, literature data so pl. 98 ^o C (V.V. Orlovsky, 1969),
isobutylphosphite S-isobutylisothiuronium with a yield of 43%, mp 162-164 ^o C.

The structure of the obtained compounds was confirmed by IR spectra - all obtained compounds have characteristic absorption bands in the region υ _PO 1195-1207 cm ^-1 , υ _CN 1680-1695 cm ^-1 , υ _NH 1568-1590 cm ^-1 , and UV spectra having a characteristic short-wave absorption with λ _max = 201-203 nm and 223-225 nm.

 Biological study of phosphorus-containing derivatives of S-alkylisothiuronium.

 This section presents the results of the initial pharmacological evaluation of phosphorus-containing derivatives of S-alkylisothiuronium. The experiments were conducted on 52 anesthetized (sodium ethaminol, 45 mg / kg, intraperitoneal) cats of both sexes weighing from 2.8 to 4.2 kg. Blood pressure was recorded using a mercury manometer and an ECG in a second standard lead. At the same time, the respiratory rate was also recorded. The severity of the hypertensive effect was assessed (as a percentage of the initial level of systolic blood pressure (SBP)), the duration of observation was 1-3 hours, the heart rate (HR) was calculated by ECG. The results are shown in FIG. 1-10 and in table. 1.

 Methylphosphite S-methylisothiuronium increases blood pressure by 20%. The hypergenic effect persists for 1 hour, the compound does not affect heart rate (Fig. 1).

 S-methylisothiuronium dimethylphosphate is one of the most active compounds; it increases blood pressure by almost 30%. The hypertensive effect persists for 1.5 hours, the compound increases the respiratory rate (NPV) (Fig. 2).

 S-ethylisothiuronium metaphosphate increases blood pressure by 25 mm Hg. Art., which is about 15% of the initial level. Hypertension affects NPV (Fig. 3).

 Ethylphosphite S-ethylisothiuronium increases blood pressure by 15%. The duration of action is 1.5 hours, the compound causes bradycardia, but does not affect the NPV (Fig. 4).

 S-ethylisothiuronium diethyl phosphate is one of the most active compounds. In the studied dose, it increases SBP by 40%, increases NPV (Fig. 5).

 Propylphosphite S-propylisothiuronium increases blood pressure by 24%. The hypertensive effect persists for 1 hour, the compound causes moderate tachycardia, but does not affect the NPV (Fig. 6).

 S-isopropylisothiuronium metaphosphate slowly increases SBP, the maximum effect is observed only after 15 minutes, but the effect persists for more than 2 hours. Does not affect NPV (Fig. 7).

 Isopropylphosphite S-isopropylisothiuronium is one of the most promising substances that increases SBP by almost 30%, the maximum effect is reached already at the 2nd minute after administration and lasts more than 2 hours. Transient bradycardia is noted, the connection does not affect the NPV (Fig. 8).

 S-butylisothiuronium dibutyl phosphate slightly increases blood pressure. The hypertensive effect persists for 1 hour (Fig. 9).

 Isobutylphosphite S-isobutylisothiuronium has a moderate hypertensive effect lasting 2 hours, while increasing heart rate and NPV (Fig. 10).

 The following are the main results of experiments confirming the possibility of using diethylphosphate S-ethylisothiuronium in the directions indicated in the description.

 Experiments on the effects of diethylphosphate S-ethylisothiuronium on systemic hemodynamics were performed on 8 anesthetized cats weighing 2-4 kg. Urethane was used for anesthesia (intraperitoneally in the form of a 30% solution at the rate of 1-1.2 g / kg). In order to identify the period of maximum effectiveness and duration of action of the drug, the main parameters of the systemic circulation were determined before its administration (initial data), as well as at 2, 15, 30, and 60 minutes after the administration of diethylphosphate S-ethylisothiuronium at a dose of 5 mg / kg.

 Systemic hemodynamics was evaluated by a combination of the following indicators: blood pressure (BP), heart rate (HR), cardiac output (IOC), cardiac systolic volume (CO), total peripheral vascular resistance (OPSS), left ventricle (Alzh) .

 Blood pressure was recorded in cats in the left common carotid artery using a mercury manometer. To prevent blood coagulation, 0.2-0.3 ml (1000-1500 IU) of heparin was injected intravenously. The heart rate was calculated by the R-R interval of the electrocardiogram, the recording of which was carried out by a two-channel electrocardiograph ELKAR-2 (model 0-87). IOC was calculated by Ramirez A. A. et al. (1956). On the basis of data on IOC, blood pressure, central blood volume and heart rate, heart CO, Alge and OPSS were calculated by the formula Wiggers C. G. (1947). The digital material obtained in the experiments was subjected to statistical processing using programs for calculating Student t-test. The results are summarized in table 2.

 The results of the experiments showed that a single intravenous administration of diethylphosphate S-ethylisothiuronium at a dose of 5 mg / kg causes a rapid and pronounced increase in blood pressure. Already at the 2nd minute after drug administration, blood pressure was 141% relative to the initial level and remained at a high level for a long time. At the 60th minute, the level of blood pressure decreased compared to that set at the 30th minute, but still exceeded (by 11%) the initial value. The difference in the values characterizing the level of blood pressure during the hour was significant and statistically significant. The severity and duration of the increase in blood pressure depended mainly on the increase in OPSS and to a lesser extent (at the 2nd minute of the drug's action) on the increase in IOC (Table 2).

 Already at the 2nd minute after the introduction of diethylphosphate, S-ethylisothiuronium OPSS was 135% relative to the initial level and remained high throughout the entire observation period. At the 60th minute, it exceeded the initial value by 58%. Therefore, the total tone of vascular resistance after drug administration increased (table. 2). IOC at the 2nd min after administration of the drug increased by 7%, and at the 15th and 30th min decreased by 11 and 22%, respectively. Such changes in the IOC were determined by the nature of the change under the influence of the drug - CTC, the magnitude of the venous return of blood to the heart, and as a consequence of this, WITH the heart. The increase in Alzh was transient, so on the 2nd and 15th min after the injection of the substance it exceeded the initial value by 89% and 22%, and by the 30th min it returned to the initial level. The results obtained indicate that the hypertensive effect that occurs with a single intravenous administration of diethylphosphate S-ethylisothiuronium and lasts for a long time (up to 1 hour), depends mainly on the increase in SPS and to a lesser extent (mainly in the initial period of the drug) IOC.

It is known that adrenomimetics practically do not suppress hypotension caused by adrenergic blockers. It was of interest to study the effect of diethylphosphate S-ethylisothiuronium with hypotension arising from the administration of the known α ₁ -adrenergic blocking agent, prazosin (adversuten). Adversuten was administered once intravenously at a dose of 1 mg / kg. The results are summarized in table 3. Based on the selective site of action of adversuten, the peripheral resistance of blood vessels and, consequently, the level of blood pressure are significantly reduced. After the administration of adverzuten already at the 5th minute, blood pressure decreases by 51 mm Hg. Art., against this background, the introduction of diethylphosphate S-ethylisothiuronium already at the 2nd minute increases blood pressure by 29 mm RT. Art. At the 15th and 30th min, the blood pressure level was 11.8 and 9.4% higher compared to the 5th min after the administration of adverzuten. No statistically significant changes in heart rate and respiratory rate were observed (with the exception of the 60th minute after the administration of diethylphosphate S-ethylisothiuronium, when bradycardia was noted). Based on the foregoing, we can conclude that diethylphosphate S-ethylisothiuronium increases blood pressure and against the background of the action of α ₁ -adrenergic blocking agents, although less long than without their purpose.

 As an experimental model of hemorrhagic shock, which is close in its course to severe clinical forms, acute blood loss (3.2% of the animal’s body weight) was selected, which is 45-47% of the circulating blood volume, over 3-5 minutes. In order to identify the maximum effectiveness and duration of the drug, the main parameters of systemic circulation were determined before blood loss, 30 minutes after bloodletting, and also at 2, 10, 30 and 60 minutes after intravenous administration of the drug. The results are shown in table 4.

 In the control group consisting of 8 animals fixed to the machine, blood loss caused a drop in blood pressure from 140.0 ± 3.9 to 65.0 ± 4.1 mm Hg. Art., which amounted to 46% of the initial level, heart rate at the 30th minute from the onset of blood loss increased by 18%. IOC decreased from 102.8 ± 4.5 to 44.0 ± 5.2 ml / min • kg, i.e. 57% compared to the original.

 The decrease in blood pressure by the 30th minute from the onset of acute blood loss was mainly due to a decrease in IOC, which, in turn, depends on a decrease in stroke volume of blood (up to 36%) and central blood volume. Thus, posthemorrhagic hypovolemia leads to a decrease in IOC, which cannot be compensated by an increase in OPSS and an increase in heart rate.

 Alge calculations indicate a decrease of 6 times.

 At the 2nd minute after the administration of the isotonic solution, the hemodynamic parameters did not differ much from the same at the 30th minute after the onset of acute blood loss, however, at the 10th and 30th minutes after the administration of 1 ml of the isotonic solution of sodium chloride, the systemic hemodynamics were progressively worsened. So, 30 min after injection of a solution of blood pressure decreased by 13% compared with the 30th min from the start of bloodletting. ChOS decreased by 7%, and IOC and CO decreased by 11 and 8%, respectively, Alzh decreased by half, and OPSS increased by 8%. A further decrease in IOC, due to a decrease in the productivity of the left ventricle, led to the death of animals.

 A single intravenous administration of diethylphosphate S-ethylisothiuronium at a dose of 5 mg / kg with hemorrhagic shock quickly and clearly increases blood pressure. So, already at the 2nd minute after the injection of the drug substance, blood pressure increased by 88% (compared to the 30th minute from the moment of blood loss) and this effect persists throughout the observation period. On the 2nd minute after the administration of diethylphosphate S-ethylisothiuronium, a significant decrease in heart rate was noted, which can be explained as a reflex effect on the heart as a result of a sharp increase in blood pressure. S-ethylisothiuronium diethylphosphate has a two-phase effect on IOC. Initially, the IOC increases by 19%, followed by its decrease by the 30th and 60th min after drug administration. However, it should be noted that in the control group with the introduction of 1 ml of isotonic solution at the 30th and 60th min, the same decrease in IOC was revealed. СО significantly increased only at the 2nd minute after the injection of the drug. Alge increased significantly at the 2nd and 10th min. OPSS significantly increased during the entire observation period.

 Analyzing the change in systemic hemodynamics under the influence of diethylphosphate S-ethylisothiuronium, we can conclude that the substance increases blood pressure in the initial period due to an increase in IOC and OPSS, and in the late due to an increase in OPS.

 The effect of diethylphosphate S-ethylisothiuronium on the background of arterial hypotension caused by the well-known ganglion blocker - hexonium was studied. Hexonium was administered once intravenously at a dose of 10 mg / kg. Sciatic and vagus nerves were stimulated by an EM-1 electric impulse above a threshold threshold rectangular current of 1 m • s duration, 10 seconds of stimulation and 20 imp / sec (peripheral section of the vagus nerve) and 300 imp / sec (central end of the sciatic nerve). The results are shown in table 5. 2 minutes after the introduction of hexonium, blood pressure decreased by 46%. The introduction of diethylphosphate S-ethylisothiuronium already at the 2nd minute increases blood pressure by 61 mm RT. Art., and a significant increase in blood pressure is maintained throughout the observation period. Heart rate has changed in the direction of bradycardia, however, these changes are statistically unreliable. Respiratory rate did not change. Ganglioblocking properties with the introduction of diethylphosphate S-ethylisothiuronium are fully preserved, as evidenced by the absence of changes in blood pressure during repeated (at 2, 15, 30 and 60 min) stimulations of the peripheral segment of the vagus and central end of the sciatic nerves.

 It can be concluded that diethylphosphate S-ethylisothiuronium quickly restores blood pressure after hypotension caused by ganglion blockers and maintains it for a long time.

крови. рНа и рН исследовали микрометодом Аструпа. Содержание гемоглобина (Нв) определяли цианогемоглобиновым методом. Газовый состав крови и рН приведены в соответствии с температурой тела животного. Содержание бикарбонатных ионов [HCO₃ ^-], показатели сдвига буферных оснований (BE) рассчитывали с помощью линейки Северинхауза. С целью упрощения расчеты проводили на ЭВМ модели GM-1403-02 по ранее составленной программе на языке Фортран. Показатели КЩС и газового состава крови исследовали в исходном состоянии, на 30-й мин с момента начала острой кровопотери, а также на 2-й, 10-й, 30-й и 60-й минутах после введения лекарственных веществ. Вещество вводили однократно внутривенно на 30-й мин с момента кровопускания.The effect of S-ethylisothiuronium diethyl phosphate at a dose of 5 mg / kg on the main indicators of acid-base condition (KHS) and blood gas composition in hemorrhagic shock was also studied in experiments on cats anesthetized with intraperitoneal administration of sodium ethaminol (nembutal) at a dose of 40 mg / kg . Blood loss was caused by bloodletting from the common carotid artery in the amount of 3.2% of the animal’s mass, which approximately corresponds to 45-47% of the circulating blood volume. Bloodletting was carried out for 3-5 minutes. Voltage indicators of oxygen and carbon dioxide in arterial (PaO ₂ , PaCO ₂ ) and mixed venous

blood. pH and pH were investigated by the Astrup micromethod. The hemoglobin (HB) content was determined by the cyanohemoglobin method. Blood gas composition and pH are given in accordance with the body temperature of the animal. The content of bicarbonate ions [HCO ₃ ^- ], the shift indicators of buffer bases (BE) were calculated using the Severinhaus ruler. To simplify the calculations, the calculations were carried out on a computer model GM-1403-02 according to a previously compiled program in Fortran. Indicators of KShchS and gas composition of blood were investigated in the initial state, at the 30th minute from the onset of acute blood loss, as well as at the 2nd, 10th, 30th and 60th minutes after the administration of drugs. The substance was administered once intravenously for 30 minutes from the moment of bloodletting.

 The control group (n = 8). Cats were injected with 1 ml of isotonic sodium chloride solution.

достоверно уменьшался с 7,284±0,011 в исходном состоянии до 7,184±0,028, 7,127±0,049 и 7,081±0,049 соответственно на 30-й, 40-й и 60-й минутах после кровопускания (p<0,05).The experimental results showed that in control cats with acute blood loss at the 30th minute after bloodletting, changes in arterial blood coronary artery syndrome were twofold, 5 out of 7 cats had metabolic and respiratory acidosis, and two had metabolic acidosis with compensated respiratory alkalosis. The pH of arterial blood decreased on average from 7.313 ± 0.015 in the initial state to 7.284 ± 0.033 by the 30th minute from the onset of blood loss. At the 40th and 60th minutes after bloodletting, a further decrease in pH was observed to 7.242 ± 0.056 and 7.152 ± 0.072, respectively. A more pronounced decrease in pH was observed on the part of venous blood. So, in 7 cats from 8 to the 30th minute after blood loss, the pH decreased, but in one did not undergo significant changes.

significantly decreased from 7.284 ± 0.011 in the initial state to 7.184 ± 0.028, 7.127 ± 0.049 and 7.081 ± 0.049, respectively, at the 30th, 40th and 60th minutes after bloodletting (p <0.05).

After acute blood loss, the metabolic parameters of the coronary artery and arterial venous blood for the entire observation period significantly decreased. The content of bicarbonate ions in arterial blood at the 30th, 40th, and 60th minutes after blood loss decreased by 25%, 27%, and 40%, respectively, compared with the initial data (P <0.01). In parallel with this, a significant decrease in BE a was found - from 6.97 ± 0.70 mmol / L in the initial state to - 10.61 ± 0.86 mmol / L at the 30th minute after blood loss (P <0.01 ) Subsequently, by the 30th and 60th minutes from the start of bloodletting, this indicator decreased by 87.7% and 143%, respectively. Similar changes in [NSO ₃ ^- ] and [BE] were detected by venous blood. So, on the 30th minute after blood loss, the content of bicarbonate ions [НСО ₃ ^- ] was 14.6% lower than the control.

 The deficiency of buffer bases significantly increased by the 30th, 40th, and 60th minutes after acute blood loss by 55%, 99%, and 130%, respectively, compared with the initial one (P <0.01). Thus, in the early stages after massive acute hemorrhage, according to our results, metabolic acidosis is the most common form of ASH disturbance. This is consistent with literature data.

что приводило к увеличению артериовенозного градиента РCО₂ в сравнении с исходным уровнем. Вполне понятно, что наблюдаемое в динамике острой кровопотери уменьшение градиента существенно ухудшало условия газообмена в легких и тканях и приводило к резкому нарастанию метаболического и респираторного ацидоза. Степень оксигенации артериальной крови в контрольной группе достаточно высока, тогда как РО₂ венозной крови при этом достоверно снижалось.At the 30th minute after acute blood loss, a decrease in PaCO ₂ and a slight increase were noted

which led to an increase in the arteriovenous gradient of PCO ₂ compared with the initial level. It is quite clear that the decrease in the gradient observed in the dynamics of acute blood loss significantly worsened the conditions of gas exchange in the lungs and tissues and led to a sharp increase in metabolic and respiratory acidosis. The degree of arterial blood oxygenation in the control group is quite high, while PO _{2 of} venous blood significantly decreased.

 The group with the introduction of diethylphosphate S-ethylisothiuronium (n = 6). The drug was administered at a dose of 5 mg / kg.

 From table 6 it follows that diethylphosphate S-ethylisothiuronium in cats with hemorrhagic shock corrects metabolic acidosis caused by blood loss, while increasing the pH of arterial blood from 7.230 ± 0.015 for the 30th minute from the moment of bloodletting to 7.252 ± 0.018 and 7.310 ± 0.064, respectively 10th and 30th minutes after injection of diethylphosphate S-ethylisothiuronium.

 Similarly to changes in pH, there were also changes in the deficiency of buffer bases, which at the 10th and 30th minutes after the administration of the drug substance significantly decreased by 20% and 18%, respectively, compared with the values observed at the 30th minute of shock (Table 6) . On average, the content of bicarbonate ions of arterial blood at the 10th and 30th min after the administration of diethylphosphate S-ethylisothiuronium increased by 5% and 16%, respectively, compared to the initial values (Table 6).

 The same changes were observed from the main indicators of the acid-base state of mixed venous blood, however, these changes were statistically unreliable.

 The partial tension of carbon dioxide in arterial and venous blood compared with the 30th minute after bloodletting decreased by 10th and 30th minutes after injection of diethylphosphate S-ethylisothiuronium. So, on the 10th minute it decreased by 9% (table. 6, table. 7). The partial pressure of oxygen in arterial and venous blood under the action of diethylphosphate S-ethylisothiuronium changed differently: in arterial blood it tended to increase, and in venous blood it decreased, but these variations were statistically unreliable.

 Analyzing the data on the effect of etrone and S-ethylisothiuronium diethylphosphate on the acid-base balance and blood gas composition under conditions of hemorrhagic shock, we can conclude that S-ethylisothiuronium diethylphosphate corrects metabolic acidosis more effectively, affecting, apparently, not only the increase in bicarbonate content ions, but also by increasing the phosphate buffer, since the molecule of diethylphosphate S-ethylisothiuronium, in comparison with etiron, contains diethylphosphate.

 We can also assume that diethylphosphate S-ethylisothiuronium affects the blood acid-base balance by freeing the body from the end products of metabolism: in particular, from lactic acid due to its more active inclusion in the measles and Krebs cycles, while maintaining the conjugation of oxidation and phosphorylation.

 In the next cycle of experiments, we studied the effect of diethylphosphate S-ethylisothiuronium on the cardiovascular system of dogs. Blood pressure was measured in two ways: recorded on the caudal artery using a piezoelectric transducer on a Ugo Bazile pressure measuring device or in the femoral artery using a catheter pre-filled with heparin. The electrocardiogram was recorded in three standard leads on a Mingograph-82.

 In FIG. 11 presents data on the selection of the optimal dose of the drug with the intravenous route of administration. S-ethylisothiuronium diethyl phosphate was administered at doses of 1 mg / kg (n = 5), 5 mg / kg (n = 18) and 100 mg / kg (n = 8). As can be seen from the drawing, diethylphosphate S-ethylisothiuronium in all doses used has a pronounced hypertensive reaction. The optimal dose is apparently a dose of 5-10 mg / kg. On the electrocardiogram, in the study of all the above doses of the drug, no pathological changes were detected. Only the development of a bradycardial reaction is noted, which develops already from the first minutes after administration, reaching an average of 25-30% of the initial level. The maximum value of this reaction is observed for 30-40 minutes after the start of the experiment and persists until the 60th minute of observation.

 Of particular note is the fact that the severity of a hypertensive reaction in dogs with low blood pressure (hypotensives) is significantly higher than in dogs with a high initial level of blood pressure (hypertensives). For this reason, further study was carried out separately for normotonics and hypotonics.

 In FIG. Figures 12 and 13 show blood pressure with the introduction of various doses of diethylphosphate S-ethylisothiuronium. As can be seen from the drawings, it was confirmed that the hypertensive reaction in dogs with low blood pressure is stronger and longer, and the optimal dose of diethylphosphate S-ethylisothiuronium with the intravenous route of administration is 5-10 mg / kg.

 In FIG. 14 shows blood pressure indicators in experiments on dogs with the oral route of administration of diethylphosphate S-ethylisothiuronium. It is shown that in this case the quantitative effect of increasing blood pressure is preserved, and the duration of the drug increases significantly.

In experiments on mice of the F ₁ line (C ₅₇ BlxCBA), the functional state of the body was evaluated by biochemical tests with the introduction of diethylphosphate S-ethylisothiuronium with the intravenous route of administration at a dose of 500 mg / kg once and at a dose of 100 mg / kg in a chronic experiment for 30 days in a row. In an acute experiment, blood was taken 30, 60, and 75 minutes after drug administration. In a chronic experiment, blood was examined daily 1 hour after drug administration. The terms of the study 1, 7, 14 and 21 days after administration of the drug. The state of protein, lipid and carbohydrate metabolism was assessed, for which the concentration of total protein, total cholesterol and glucose in the blood serum of mice was determined by conventional methods. Total protein was determined by the biuret method, total cholesterol was determined by the Ilke method, and glucose by the oxidase method.

 Blood was obtained from mice after rapid decapitation, the test material was taken at the same time of the day in the same season of the year.

Blood to obtain serum was collected in centrifuge tubes, placed in a thermostat for 30-40 minutes, the resulting clot was circled with a needle, then placed in a refrigerator for 1 hour at a temperature of + 4 ^o С, after cooling, it was centrifuged at 3000 rpm for 10-15 min The resulting serum with no signs of hemolysis was collected in a clean tube. All quantitative indicators of blood biochemical characteristics were obtained using an SF-26 spectrophotometer. Calibration curves were previously constructed to determine total protein and total cholesterol. The data obtained are summarized in tables 8 and 9.

 As a result of the study, it was found that after the administration of diethylphosphate S-ethylisothiuronium at a dose of 500 mg / kg, there were significant increases in the concentration of total protein, cholesterol and glucose after 30 minutes, which later tended to normalize.

 Due to the fact that the liver is included in all biochemical processes that occur in the body and is the site of the formation of protein components of the blood, it can be assumed that diethylphosphate S-ethylisothiuronium at a dose of 500 mg / kg affects the functional state of the liver. An increased level of total protein is possibly associated with a violation of the regulatory processes of the synthesis of blood protein by the liver, as well as damage to the parenchyma.

 Thus, we can conclude that the introduction of diethylphosphate S-ethylisothiuronium at a dose of 500 mg / kg leads to a change in indicators of protein, lipid and carbohydrate metabolism, which is obviously associated with a lack of compensatory capabilities of the liver and a decrease in functional abilities. These changes go beyond physiological adaptive reactions and lead to changes lying on the verge of pathology, however, these changes are not accompanied by long-term changes in the functional state, the changes are reversible.

 Under the conditions of a chronic experiment, it was shown that at a dose of 100 mg / kg of diethylphosphate S-ethylisothiuronium, the fluctuations of the studied parameters do not go beyond physiological fluctuations and do not cause the observed changes from biochemical parameters.

 In experiments on dogs, the functional state of the body was evaluated by biochemical tests with the introduction of diethylphosphate S-ethylisothiuronium in the form of a 10% solution for injection intravenously at a dose of 10 mg / kg daily at the same time for 3 consecutive days. Blood was examined daily 1 hour after drug administration for 3 days and after administration on days 4, 5, and 10. The state of protein, lipid and carbohydrate metabolism was assessed, for which the concentration of total protein, albumin, globulin, total cholesterol and glucose in the blood serum of dogs was determined by conventional methods. Total protein was determined by the biuret method, total cholesterol was determined by the Ilke method, and glucose by the oxidase method. The results are summarized in tables 10-14.

 After administration of the drug, the level of total protein had wave-like fluctuations with a tendency to decrease on the 4th and 5th day, the level of albumin and globulins also wave-like, the level of total cholesterol and glucose had insignificant fluctuations that did not go beyond the limits typical for healthy dogs .

 Thus, based on a study of the general toxic effect and biochemical parameters in a daily three-day experiment on the intravenous administration of a drug at a dose of 10 mg / kg in the form of a 10% solution, we can conclude that the drug, in addition to a specific antihypertensive effect, does not have a significant effect on the health status of dogs, there were no signs of intoxication or any significant deviations of the studied parameters compared to the initial data, individual sensitivity of animals was not noted x for intravenous administration of the drug.

 As a result of a pathomorphological study of the internal organs of experimental animals (mice, dogs), subjected to single and repeated exposure to diethylphosphate S-ethylisothiuronium in moderate lethal and sublethal doses, it was found that this drug in the conditions of the experiments causes mild signs of hemodynamic disturbance in the early stages of observation in the form of expansion of the gaps and plethora of mainly small and medium vessels of the arterial and venous channels, mainly in the liver, m okarde, lung, spleen, and to a lesser extent, in the kidney. The circulatory disorder is reversible and, as a rule, normalizes after 7-10 days after the termination of the course of exposure to the drug. No marked inflammatory, dystrophic, or necrobiotic changes on the part of internal organs were observed in all experiments. In a number of cases, signs of compensatory-adaptive reactions from the liver were noted (proliferation of Kupffer cells, hyperchromia of the cytoplasm and nuclei of hepatocytes).

Claims (3)

1. A tool that affects blood pressure, containing as an active substance a phosphorus-containing derivative of S-alkylisothiuronium of the General formula

where Alk is a branched or unbranched C ₁ -C ₄ alkyl;
x ^- is the anion of phosphorus-containing acid.  2. The tool according to claim 1 for the treatment of emergency conditions in the form of ampoule solutions containing 1-10% of the active substance.  3. The tool according to claim 1 for the treatment of hypotonic conditions in the form of solid dosage forms at a dose of the active substance of 0.1-20 mg / kg

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