RU2414453C2 - 1-alkyl-2-alkylcarbamoyl-glycerins exhibiting antihypertensive activity, and method for preparing thereof - Google Patents

Abstract

FIELD: medicine, pharmaceutics. ^ SUBSTANCE: invention refers to medicine, namely to an agent exhibiting antihypertensive activity which represents 1-alkyl-2-alkylcarbamoylglycerins of general formula I . In formula I R means hydrocarbon radical -(CH2)nCH3 (n=10-18), R1 means methyl or ethyl. Also, the invention refers to a method for preparing the compounds of formula I. The method consists in the fact that parent 1-alkylglycerins of general formula II react with trimethylchlorosilane with using triethylamine in toluene medium at temperature -20C to 0C, then the reaction mass is added with appropriate alkylisocyanate and processed with ammonium bifluoride in methanol medium at room temperature. ^ EFFECT: preparation of the agent exhibiting antihypertensive activity. ^ 2 cl, 7 ex

Description

The invention relates to chemistry and medicine, namely to antihypertensive agents and methods for their preparation.

BACKGROUND

The closest analogues in structure and biological activity are 1-alkyl-2-acetyl-glycerins of the formula

where R is a hydrocarbon radical With ₁₆ -C ₁₈ ,

which exhibit marked antihypertensive activity, but are unstable due to the tendency of the acetyl group to migrate from position 2 of the glycerol molecule to position 3, which leads to a loss of physiological activity within 10-30 min after their introduction into the bloodstream [1-5].

As the starting 1-alkyl-glycerins, compounds of the general formula [6, 7] were used:

where R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18).

The technical result to which this invention is directed is to obtain new derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing in the 2nd position of the molecule an alkyl carbamoyl group and having pronounced prolonged antihypertensive activity and a method for their preparation.

SUMMARY OF THE INVENTION

1. The technical result is achieved by the fact that proposed 1-alkyl-2-alkylcarbamoyl-glycerins, of the General formula

where a) R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18), R 'is methyl;

b) R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18), R 'is ethyl,

possessing antihypertensive activity.

The present invention is represented by the chemical structure of the new compounds in the series of 1-alkyl-glycerols, namely 1-alkyl-2-alkylcarbamoyl-glycerols containing in the 2nd position of the molecule an alkylcarbamoyl group of the general formula

where a) R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18), R 'is methyl;

b) R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18), R 'is ethyl,

possessing antihypertensive activity.

The inventive compounds are crystalline substances of white color, insoluble in water, readily soluble in ethanol, methanol, a mixture of methanol and chloroform. The structure and identity of the claimed compounds are confirmed by elemental analysis, ¹ H NMR and IR spectra and thin layer chromatography.

2. The technical result is also achieved by the fact that the proposed method for producing 1-alkyl-2-alkylcarbamoyl-glycerols containing in the 2nd position of the molecule alkylcarbamoyl group and having a pronounced antihypertensive activity, in which the original 1-alkyl-glycerols are reacted with trimethylchlorosilane in the presence of triethylamine in toluene at -20 ÷ 0 ° C, then with an alkyl isocyanate and subsequent treatment of the reaction mixture with ammonium bifluoride in methanol at room temperature, as the starting 1-alkyl-hl compounds of the general formula used

where R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18).

The proposed method consists in the fact that the starting 1-alkyl-glycerols are reacted with trimethylchlorosilane in the presence of triethylamine in toluene at -20 ÷ 0 ° C, then with an alkyl isocyanate and then the reaction mixture is treated with ammonium bifluoride in methanol at room temperature.

Compounds of the general formula are used as starting 1-alkyl-glycerins

where R is the hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18).

Example 1. Obtaining 1-hexadecyl-2-methylcarbamoyl-glycerol.

To a solution of 3.67 g (0.012 mol) of 1-hexadecylglycerol and 1.17 g (0.012 mol) of triethylamine in 150 ml of toluene in an argon atmosphere at -20 ÷ 0 ° С, a solution of 1.26 g (0.012 mol) of trimethylchlorosilane in 20 ml of toluene is added. The reaction mixture was kept at 0 ° C, 2 ml of methyl isocyanate was added and left overnight. Then it is filtered, the solvent is removed in vacuo, the residue is dissolved in 40 ml of hexane, 0.66 g (0.012 mol) of ammonium bifluoride in 40 ml of methanol are added, and the mixture is kept at room temperature for 4 hours. The mixture was filtered through an alumina layer, the solvent was removed in vacuo, 150 ml of hexane was added, filtered, hexane was removed in vacuo to give 3.33 g (77%) of 1-hexadecyl-2-methylcarbamoyl-glycerol), mp 59 ° C, R _f = 0.3 (silica gel, ether-hexane = 4: 1). 1 H NMR (δ, ppm): 0.9 (3H, t) - CH ₃ ; 1.2 (28 N, s.)

(-CH ₂ -) ₁₄ ; 1.6 (2H, m) OS H ₂ (CH ₂ ) ₁₄ CH ₃ ; 2.8 (3H, s); CONHCH ₃ ; 3.0 (1H, O) —CH ₂ C H CH ₂ ; 3.4-3.6 (4H, m) -CH ₂ CHCH ₂ -; 3.8 (1H, s) CH ₂ O H ; 4.9 (1H, m) OCONHCH ₃ . IR spectrum (ν _max , cm ^-1 ): 1700 (СО); 3300 (NH, OH). Found,%: C 67.05; 67.12; H 11.45; 11.13; N, 2.85; 2.94. C ₂₁ H ₄₃ NO ₄ . Calculated,%: C 67.56; H 11.53; N, 3.75.

New derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of the alkylcarbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity are obtained.

Example 2. Obtaining 1-hexadecyl-2-ethylcarbamoyl-glycerol.

To a solution of 3.67 g (0.012 mol) of 1-hexadecylglycerol and 1.17 g (0.012 mol) of triethylamine in 150 ml of toluene in an argon atmosphere at -20 ÷ 0 ° С, a solution of 1.26 g (0.012 mol) of trimethylchlorosilane in 20 ml of toluene is added. The reaction mixture was kept at 0 ° C, 2 ml of ethyl isocyanate was added and left overnight. Then it is filtered, the solvent is removed in vacuo, the residue is dissolved in 40 ml of hexane, 0.66 g (0.012 mol) of ammonium bifluoride in 40 ml of methanol are added, and the mixture is kept at room temperature for 4 hours. The mixture was filtered through a layer of alumina, the solvent was removed in vacuo, 150 ml of hexane were added, filtered, hexane was removed in vacuo to give 3.8 g (85%) of 1-hexadecyl-2-ethylcarbamoyl-glycerol), mp 50 ° C, R _f = 0.34 (silica gel, ether-hexane = 4: 1). 1 H NMR (δ, ppm): 0.9 (3H, t) -C H ₃ ; 1.2 (28 N, s.) (-C H ₂ -) ₁₄ ; 1.6 (2H, m) -C H ₂ OC ₁₆ H ₃₃ ; 2.4 (2H, s) -CONHC H ₂ CH ₃ ; 3.4 (2H, s) -NHC H ₂ CH ₃ ; 3.6 (4H, m) - CON H CH ₂ CH ₃ ; 5.2 (1H, m) -CH ₂ C H CH ₂ . Found,%: C 67.91; 67.88; H 10.82; 10.91; N 3.10; 3.24. C ₂₂ H ₄₅ NO ₄ . Calculated,%: C 68.21; H, 11.62; N, 3.62.

New derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of the alkylcarbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity are obtained.

Example 3. Obtaining 1-octadecyl-2-methylcarbamoyl-glycerol.

To a solution of 7.74 g (0.02 mol) of 1-octadecylglycerol and 2.02 g (0.02 mol) of triethylamine in 200 ml of toluene in an argon atmosphere at -20 ÷ 0 ° С, a solution of 3.01 g (0.02 mol) of trimethylchlorosilane in 40 ml of toluene is added. The reaction mixture was kept at 0 ° C, 4 ml of methyl isocyanate was added and left overnight. Then it is filtered, the solvent is removed in vacuo, the residue is dissolved in 60 ml of petroleum ether, 1.1 g (0.02 mol) of ammonium bifluoride in 60 ml of methanol are added, and the mixture is kept at room temperature for 4 hours. The mixture was filtered through a layer of alumina, the solvent was removed in vacuo, 200 ml of petroleum ether was added, filtered, the solvent was removed in vacuo to give 5.65 g (65%) of 1-octadecyl-2-methylcarbamoyl-glycerol), mp 46 ° C, R _f = 0.42 (silica gel, ether-petroleum ether = 3: 1). PMR (δ, ppm): 0.9 (3H, t) -CH ₃ ; 1.2 (30 N, s.) (-CH ₂ -) ₁₅ ; 1.6 (2H, m) OC H ₂ (CH ₂ ) ₁₅ C H ₃ ; 2.8 (3H, s); CONHCH ₃ ; 3.0 (1H, O) -CH ₂ C H CH ₂ ; 3.4-3.6 (4H, m) -C H ₂ SNA H ₂ -; 3.8 (1H, s) CH ₂ O H ; 4.9 (1H, m) OSONSN ₃ . Found,%: C 67.81; 67.75; H 11.00; 11.21; N 3.10; 3.04. C ₂₂ H ₄₅ NO ₄ . Calculated,%: C 68.21, H 11.62, N 3.62.

New derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of the alkylcarbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity are obtained.

Example 4. Obtaining 1-octadecyl-2-ethylcarbamoyl-glycerol.

To a solution of 4 g (0.01 mol) of 1-octadecylglycerol and 1 g (0.01 mol) of triethylamine in 150 ml of toluene in an argon atmosphere at -20 ÷ 0 ° С, a solution of 1.05 g (0.01 mol) of trimethylchlorosilane in 20 ml of toluene is added. The reaction mixture was kept at 0 ° C, 2 ml of ethyl isocyanate was added and left overnight. Then it is filtered, the solvent is removed in vacuo, the residue is dissolved in 40 ml of petroleum ether, 0.55 g (0.01 mol) of ammonium bifluoride in 40 ml of methanol are added and incubated for 4 hours at room temperature. The mixture was filtered through a layer of alumina, the solvent was removed in vacuo, 100 ml of petroleum ether was added, filtered, the solvent was removed in vacuo to give 3.06 g (68%) of 1-octadecyl-2-ethylcarbamoyl-glycerol), mp 42 ° C, R _f = 0.44 (silica gel, ether-petroleum ether = 3: 1). 1 H NMR (δ, ppm): 0.9 (3H, t) - C H ₃ ; 1.6 (30H, s.) (-CH ₂ -) ₁₅ ; 1.6 (2H, m) - С Н ₂ ОС ₁₆ Н ₃₃ ; 2.4 (2H, s) -CONHC H ₂ CH ₃ ; 3.4 (2H, s) -NHC H ₂ CH ₃ ; 3.6 (4H, m) - CON H CH ₂ CH ₃ ; 5.2 (1H, m) -CH ₂ C H CH ₂ . Found,%: C 67.96; 68.11; H 11.00; 11.11; N, 3.30; 3.26. C ₂₃ H ₄₇ NO ₄ . Calculated,%: C 68.82, H 11.72, N 3.49.

New derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of the alkylcarbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity are obtained.

The study of the synthesized compounds for lowering blood pressure was carried out in rats with vasorenal hypertension and in rats with spontaneous hypertension, as shown in examples 5, 6, 7.

Example 5. The study of synthesized compounds to lower blood pressure.

In the following experiments on laboratory animals, in particular rats, to obtain vasorenal hypertension, the Goldblatt model was used - “1 clip - 2 kidneys” or “1 clip - 1 kidney”. The application of a clip restricting blood flow to the renal artery leads to activation of the renin-angiotensin-aldosterone system and to an increase in mean arterial pressure (BP). The degree of increase in blood pressure depends on the diameter of the clip. The “1 clip - 1 kidney” model involves the removal of one kidney. In this case, the initial rise in blood pressure is also due to the activation of the renin-angiotensin-aldosterone system, the indicators of which return to normal after some time. Moreover, the situation is aggravated by the development of renal failure, the accumulation of excess fluid in the body, and in the later stages of the development of the disease, increased blood pressure is supported by other mechanisms.

Male Wistar rats with an initial weight of 240-300 g were used in the experiment. A pre-anesthetized rat — sodium ethamine 40 mg / kg, intraperitoneally — was placed on the operating table in the “on the back” position with limb fixation. The surgical suture was treated with streptocide. Arterial systolic pressure was measured in the caudal artery in waking rats. Before measurement, the rat was placed in a box with a thermostatically controlled substrate with a temperature of 37 ° C for 10-15 minutes. A cuff was put on the base of the tail of the rat, connected through a buffer tank to a pressure gauge and a rubber bulb. A sensor was located near the cuff, recording changes in the tail circumference during the passage of a pulse wave. The sensor is a rubber tube filled with carbon powder. The signal from the sensor was amplified and recorded on an oscilloscope having an afterglow screen. Air was pumped into the cuff to a pressure value at which the blood flow in the caudal artery completely stopped. Then the air pressure in the cuff was slowly reduced and the level of pressure at which blood pulsations appeared was noted. This cuff pressure value corresponded to the systolic blood pressure of the animal.

The experiments on awake rats were carried out as standard: one experimental rat and one control were placed in blood pressure measuring devices, where the rat was able to move freely, a catheter was connected to a pressure transducer, and the rats were left for 1 hour to adapt to the experimental conditions. After 1 hour, the initial parameters were recorded for 20 minutes. The test preparations based on the obtained compounds were administered orally, once, in olive oil, to a experimental animal at a dose of 5 mg / kg in a volume of 0.3-0.4 ml, depending on the weight of the rat. Control rats received olive oil in an equivalent volume. Since, according to preliminary data, it was known that substances had a long-lasting effect, blood pressure was recorded intermittently, parameters were recorded every 20 min for one hour and during 10 hours of daylight hours on the first and second days of the experiment. One month after the operation to simulate vasorenal hypertension “1 kidney - 1 clip”, animals were obtained with a spectrum of changes in blood pressure. In each group there were at least 10 animals. It is known that simple removal of the kidney in rats does not lead to an increase in blood pressure, the second kidney usually copes with the increased load. This "variability" of the reaction of animals in response to the standard procedure for removing one kidney and clipping the lumen of the artery of the second renal artery may be due to several reasons. Firstly, animals have individual characteristics of the diameter of the renal artery, for some rats the application of a standard diameter clip may be critical, for others it is insufficient for the development of hypertension. Secondly, it is possible that each animal has its own internal, individual pressure range, in which the kidney copes with the load, so the pressure rise turns out to be small. Thirdly, there may be errors during the operation, for example, the shoulders of the clips are not tight enough and the diameter of the vessel in this case may be slightly larger than was intended. In a further experiment to study the effect of oral administration of test drugs on blood pressure, animals with an average pressure above 145 mm Hg were selected. Oral administration of olive oil to control animals did not lead to a change in mean blood pressure either on the first or second day of the experiment. Feeding with drugs caused a decrease in blood pressure, which by the second hour of the first day of the experiment was about 20 mm Hg. On the second day, this drop was significant, blood pressure decreased by 50-55 mm Hg, starting from the 4th hour of the experiment.

Studies have shown that all synthesized compounds with oral administration once and gradually and for a prolonged, at least 2 days, the period reduces blood pressure.

Example 6. The study of synthesized compounds to lower blood pressure.

In Okamoto and Aoki rats, the development of hypertension is genetically determined and this line is a model of primary hypertension, or human hypertension. 2 days before the experiment, animals under anesthesia received 500 mg / kg of urethane and 70 mg / kg of ∝-chloralosis, and polyethylene catheters were implanted intraperitoneally into the spinal aorta through the femoral artery to record blood pressure (BP) and heart rate (HR) and inferior vena cava through the femoral vein for the administration of the test drug. Registration of these hemodynamic parameters was carried out on awake animals in free behavior, as described above. Rats were divided into 2 groups: experimental animals, which were injected with drugs dissolved in liposomes (1 mg / ml), and control, which was administered only liposomes. The experiment began after adaptation of the animal to the experimental setting for at least 1 hour. The preparations obtained on the basis of the proposed compounds were administered by infusion at a dose of 1 mg / kg for 30 seconds. Statistical data processing was performed using the non-parametric Mann-Whitney test to compare the control and experimental groups and the non-parametric Wilcoxon criterion for paired samples for comparison with the initial background values. The synthetic metabolites of the platelet aggregation factor caused a marked decrease in blood pressure, which reached a maximum approximately 50 minutes after drug administration. Within 1 hour after administration of the drug, the average blood pressure in the experimental group dropped as much as possible on average by 26 ± 5 mm Hg. (by 17 ± 3%, p <0.05) compared with the initial background values of mean blood pressure (151 + 4 mm Hg) and was lower than the average blood pressure in rats of the control group, by 28 mm Hg. (18%; p <0.05).

Along with a decrease in blood pressure, drug administration caused tachycardia, which reached a maximum approximately 15 minutes after drug administration. Heart rate increased 1.4 times (p <0.05) compared to baseline values and was higher than heart rate in rats of the control group by 1.3 times (p <0.05). Blood pressure and heart rate in control rats did not significantly change compared to baseline values (152 + 3 mmHg and 313 + 11 beats / min, respectively). The mean blood pressure remained reduced one day after drug administration by 23 ± 3 mm Hg. (by 15 ± 2%; p <0.05) compared with the initial background values and by 23 mm Hg. (15%; p <0.05) compared with rats of the control group. Signs of tachycardia after 1 day were completely absent. The antihypertensive effect of the drugs persists for 1 day and completely disappears 4 days after its administration.

Example 7. The study of synthesized compounds to lower blood pressure.

The following experiments were performed on a segment of a perfused isolated rat tail caudal artery. The prepared segment of the rat caudal artery, fixed at one end on the cannula, was placed in the working chamber, the cannula was connected to the internal duct system and pressure sensor for internal perfusion of the vessel and measurement of perfusion pressure, respectively. Within 30 minutes, the vessel was adapted to the experimental conditions. After 30 minutes, the presentation of norepinephrine (HA) in increasing concentrations (from 10 ^-8 M to 10 ^-8 M) began, the concentration of HA that caused an increase in perfusion pressure by 100-120 mm Hg was considered working, in this solution acetylcholine (AH) and synthesized preparations were diluted. After reaching a plateau of the reaction to HA, in order to ensure the preservation of the vascular endothelium, perfusion concentration AX in

10 ^-8 M. AH, acting on vascular endothelial cells, leads to the release of nitric oxide and relaxation of the vessel. The reaction to AX was washed with the same concentration of HA. After reaching a reaction plateau, perfusion with a 10 ^-4 M solution of the tested drugs was started, then HA and saline were perfused, the experiment was completed.

In control experiments, in response to perfusion of the solvent of the preparations, dimethyl sulfoxide, no changes in the vessel tone were observed, the tested drugs in increasing concentrations caused dose-dependent relaxation to 20% of its initial level. Evidence suggests that drugs can act directly on the vessel, causing it to relax.

The preparations presented, in particular in the examples, obtained on the basis of the above compounds described in the claims, were tested for toxicity at a dose of 10 to 100 mg per 1 kg of weight in Wistar rats. In the experiment, 100 male rats weighing 240-300 grams were used. The drugs showed no toxic effect and did not affect the behavior of animals.

A study of the synthesized preparations based on the above compounds for lowering blood pressure was performed on rats with vasorenal hypertension and on rats with spontaneous hypertension, Okamoto and Aoki line.

All tested preparations based on the above compounds showed no toxicity in rats at a dose of 100 mg / kg or less.

All tested preparations based on the above compounds steadily - within 24 hours or more - reduced blood pressure with a single oral administration and when administered intravenously in liposomes, as in rats with vasorenal hypertension "1 kidney - 1 clip" in the Goldblatt modification - experimental model symptomatic human hypertension, and in rats with spontaneous hypertension - the Okamoto and Aoki line - an experimental model of human hypertension.

From the above it follows that drugs based on the proposed compounds have a prolonged antihypertensive effect with a maximum reaction on the 2nd day after administration and do not significantly affect the frequency of cardiovascular contractions.

Experiments on an isolated rat tail caudal artery showed that the antihypertensive properties of the drugs may be due to a direct effect on the vessel wall.

Thus, the desired technical result was achieved, namely, new derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of an alkyl carbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity, and a method for their preparation were obtained.

INDUSTRIAL APPLICABILITY

The invention relates to chemistry and medicine, namely to antihypertensive agents and methods for their preparation.

New derivatives of 1-alkyl-2-alkylcarbamoyl-glycerols containing molecules of the alkylcarbamoyl group in the 2nd position and having pronounced prolonged antihypertensive activity, and a method for their preparation are proposed.

Compounds obtained on the basis of the proposed inventions can be used as a prolonged-acting antihypertensive agent, as drugs for the treatment of hypertension caused by kidney disease, and hypertension, diseases that affect up to 20% of the population, mainly in industrialized countries.

INFORMATION SOURCES

1. Blank ML, Cress EA, Snyder F. "A new class of antihypertensive neutral lipid: 1-alkyl-2-acetyl-sn-glycerols, a precursor of platelet activating factor," Biochemical and biophysical Research Communications, 118, No. 1 , R. 344-350, 1984.

2. Blank ML, Spector AA, Kaduce TL, Lee T., Snyder F. "Metabolism of platelet activating factor (1-alkyl-2-acetyl-sn-glycero-3-phoshocholine) and 1-alkyl-2-acety- sn-glycerol by numan endothelial cells, "Biochimica et Biophysica Acta, 876, p. 373-378, 1986.

3. Kulikov VI, Muzya GI "The biological role of cellular metabolites - structural analogues of platelet activation factor," Biochemistry, 61, No. 3, pp. 387-403, 1996.

4. Sezdarevich B., "Glyceride isomerizations in lipid chemistry," J. Am. Oil Chem. Soc. 44, p. 381-393, 1967.

5. Van Lohuizen O.E., Varkade P.E., "Migration of acyl group in glycerol," Rec.trav chim., 79 (2), p. 133-138, 1960.

6. Malrin, T. "The synthesis of glycerides. Progress in the chemistry of fat and other lipids," 4, p. 64-67, 1957.

7. Mattson F.H., Volpenheim R.A., "Synthesis and properties of glycerids," J. Lipid Res., 3, p. 281-296, 1962.

Claims (2)

1. An agent having antihypertensive activity, representing 1-alkyl-2-alkylcarbamoylglycerols of the general formula I

,
where R is a hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18), R ¹ is methyl, ethyl. 2. The method of obtaining 1-alkyl-2-alkylcarbamoylglycerols of the General formula I

,
where the values of R and R ¹ are indicated in claim 1, according to which the starting 1-alkylglycerols of the general formula II

where R is the hydrocarbon radical - (CH ₂ ) _n CH ₃ (n = 10-18) is reacted with trimethylchlorosilane using triethylamine in toluene at a temperature of from -20 to 0 ° C, then the corresponding alkyl isocyanate is added to the reaction mass and treated ammonium bifluoride in methanol at room temperature.