RU2201234C2 - Polymeric water-soluble analog of acetyl-salicylic acid and method of its synthesis - Google Patents

Abstract

FIELD: organic chemistry, chemical-pharmaceutical industry, pharmacy. SUBSTANCE: invention relates to preparing water-soluble preparations of acetylsalicylic acid. Invention proposes novel water-soluble analog of acetylsalicylic acid of the formula:

Description

 The invention relates to the pharmaceutical industry, is associated with the production of water-soluble preparations of acetylsalicylic acid (ASA), related to non-steroidal anti-inflammatory drugs and having anti-thrombotic, analgesic, anti-inflammatory and antipyretic effects, the main of which is the effect on the adhesion and aggregation of platelets I. B [ ., Ivleva A.Ya. and other Cardiology, 8, 1998, p. 84-96]. In doses from 120 to 325 mg / day, as well as 75 and 50 mg / day, ASA is used to prevent thrombosis and embolism, with unstable angina, myocardial infarction, cerebrovascular accident, etc. [Nasonov, Russian honey. Journal, HT 2000, pp. 1-8.]. It is common to use ASA as an analgesic, anti-inflammatory, antipyretic agent in the treatment of rheumatic diseases, neurologies, migraines, etc. At the same time, this drug has side effects that limit its use. Among the most dangerous toxic phenomena, damage to the gastrointestinal tract in the form of ulcers and bleeding even in the conditions of short-term administration of reduced doses of ASA [Bokarev IN, Ivleva A.Ya. and other Cardiology, 8, 1998, pp. 84-96].

 Closest to the claimed polymer water-soluble analogue of acetylsalicylic acid is the preparation of acetylsalicylic acid, obtained by the method associated with the use of poly-N-vinylpyrrolidone (PVP) and polyethylene glycol (PEG) as complexing polymer components [patent 2054939, RU, 1996]. According to the known method, the preparation of acetylsalicylic acid (RA) was obtained by joint grinding and mixing under conditions of mechanochemical interaction of the substance ASA with a mixture of medical low molecular weight PVP polymers (m.m. 12600) and PEG (m.m. 4000-6000). The total mass content of polymers in the mixture is up to 35%; the ratio of the mass fractions of polymers is from 1: 9 to 9: 1. The choice of polymers is due to their use in the manufacture of medicines as excipients. The ASA preparation is obtained by joint mechanical grinding and mixing in an S-1266-00 jet mill at an air pressure of 0.5 MPa at the inlet of the chamber and a feed rate of 10 g / min. During this process, the mechanochemical interaction of PVP, PEG, and ASA takes place, resulting in the formation of complexes of ASA with polymers, which reports a significant increase in the rate of transition of the substance of ASA into an aqueous solution. The resulting powdery preparation is packaged in 0.77 g with a content of 0.5 g ASA. In the manufacture of tablets, starch and magnesium stearate are used as additional auxiliary substances. The rate of transition of ASA into an aqueous solution from tablets (made by a known method) is 7 times higher than that of tablets made without the use of mechanochemical transformations in the presence of PVP and PEG. The solubility of the substance ASA from the composition of tablets made by a known method is comparable with the same indicator for acetylsalicylic acid.

 The ASA preparation obtained by a known method (RA) was investigated in comparison with the ASA substance (FS 422688-94, Yilin Pharmaceutical, China) in order to evaluate antithrombotic activity and ulcerogenic effect.

 When determining antithrombotic activity, a daily dose of RA containing 75 mg of ASA was used; the reference drug (ASA) was prescribed in the same dose. An experimental study on animals showed that by day 30 there was a significant (p <0.01) decrease in the coagulation index compared with the control group of animals when administered by mouth in RA solutions by 44%, ASA by 32% RA and ASA in the prophylactic doses of 75 mg / day have comparable antithrombotic activity.

 Pathomorphological studies of the internal organs of animals showed that signs of irritation of the mucous membrane were found in the stomachs, in particular, traces of increased secretion of mucus, especially in the neck of the fundus glands, and deposition of mucus on the surface of the gastric mucosa were visible. In some places, desquamation of the integumentary epithelium is observed, which indicates ulcerogenicity of the RA.

 Pathomorphological changes in the stomachs in rats treated with the same ASA substance in the same dose were similar to the changes caused by the RA preparation. Thus, RA and ASA at a dose of 75 mg / day in experimental animals by the 30th day of administration cause damage to the gastric mucosa and renal parenchyma. The result of the comparative study is framed in the form of relevant acts attached to the description of the invention.

 The claimed invention is aimed at obtaining a polymer of a water-soluble analogue of acetylsalicylic acid, which has increased analgesic, anti-inflammatory, antipyretic, thrombolytic activity and reduced gastro- and nephrotoxicity.

 The essence of the invention lies in the fact that the polymer water-soluble form of ASA is obtained by chemical immobilization of a substance on a copolymer (CPL) of N-vinylpyrrolidone with N- (glycidyl) vinyl-γ-aminobutyric acid to form drug-polymer covalent bonds. CPL of N-vinylpyrrolidone with N- (glycidyl) vinyl-γ-aminobutyric acid containing 30-70% of units with the structure of N- (glycidyl) vinyl-γ-aminobutyric acid is obtained by treating PVP with chloromethyloxyran (EPH) in the presence of a catalyst, while the pyrrolidone cycles are opened in part of the PVP units with the formation of the structure of N-vinyl-γ-aminobutyric acid and the formation of side chains carrying reactive groups such as glycidyl or haloalkyl structures, which provide increased chemical activity in comparison with PV P. Such side chains act as spacers after chemical addition of the drug component. The structural features of the resulting SPLs determine their high hydrophilicity with a significant mass content of fixed ASA. The best option is to use as a polymer matrix SPL composition from 2: 1 to 1: 2 with the formation of a water-soluble polymer form containing chemically covalently bound ASA up to 39 wt.%.

 The results of the studies showed that the inventive polymer water-soluble analogue of ASA (IASA) has an analgesic activity that is 2 times superior in strength to ASA, which is associated with the potentiating action of the polymer matrix, anti-inflammatory, antipyretic activity is comparable to ASA substance. Pharmacological tests of IASC were carried out in doses from 1 to 100 mg / kg and included studies of analgesic, anti-inflammatory, antipyretic activity, effects on the blood coagulation system, its morphological composition and biochemical parameters of blood serum. Gastro-, nephrotoxicity and acute toxicity were evaluated in comparison with ASA in equivalent doses for the substance. Analgesic activity was determined using three generally accepted methods: 1) "vinegar cramps" (intraperitoneal injection of 3% acetic acid), 2) electric pain irritation of the paws, 3) tail compression. As a result of the studies, it was found that IASK at a dose of 100 mg / kg of animal body weight when administered orally by analgesic activity exceeds ASA by 1.5-2 times.

 According to the effect on the blood coagulation system, the claimed polymer water-soluble analogue of ASA is 1.5-2 times greater than the substance ASA in action on blood coagulation processes (in an equivalent dose of 75 mg / day). A characteristic feature of the inventive polymer water-soluble analogue of ASA is that it does not cause changes in the gastric mucosa, does not have a damaging effect when introduced to the stomach and kidneys. At the same time, ASA and RA [patent 2054939, RU, 1996] in equivalent doses of the substance ASA with a similar method of administration causes a damaging effect on the gastric mucosa and a slight degree of hydropic damage to the renal tubule epithelium.

 The acute toxicity of the inventive polymer water-soluble analogue of ASA is comparable with the acute toxicity of acetylsalicylic acid.

 Example 1

A. 20 g of EPH, 12 ml of ethanol, 0.04 ml of acetyl chloride were introduced through a dropping funnel into a solution of 6.0 g of PVP in 75 mg of water with stirring; the mixture was thermostated at 50-70 ^o C for 4 hours. The residue after evacuation of the reaction mass was extracted with ether and dried in vacuum.

3,7 г СПЛ N-винилпирролидона с N-винил-N-глицидил-γ-аминомасляной кислотой (1:1), полученного модифицированным известным способом [Мусин Р.И., Ли В. А. и др. ХФЖ, 11, с.1379-1381, 1989], 2,2 г АСК и 0,02 мл ацетилхлорида в 40 мл этанола перемешивали при 50^oС в течение 60 минут и при 70^oС 240 минут; раствор вакуумировали при 70^oС (-7 кПа). Полученное сиропообразное вещество экстрагировали ацетоном и эфиром, остаток вакуумировали в тех же условиях до образования затвердевшей бесцветной пены, после измельчения которой выделен порошкообразный продукт, растворимый в воде, спирте и хлороформе с т.пл. 120-125^oС. Выход 2,6 г (43%). Найдено, %: N 5,91. (С₆Н₉NO₇)₅(С₁₈Н₂₂NO₇)₅. Вычислено, %: N 6,13. Содержание иммобилизованной АСК, определявшееся титрованием раствора СПЛ в спирте водным раствором КОН, а также методом ИК-спектрального анализа, составляет 39,3 мас.%.The isolated amorphous solid product (5.8 g) was dissolved in 70 ml of ethanol containing 1.2 KOH. The solution was thermostated at 25 ^o C for 4 hours, the residue was filtered on a KCl-1.4 filter, the filtrate was evacuated, the residue (4.1 g) was a solid water-soluble product, a copolymer of N-vinylpyrrolidone and N-vinyl-N-glycidyl-γ-aminobutyric acids (1: 1)

3.7 g CPF N-vinylpyrrolidone with N-vinyl-N-glycidyl-γ-aminobutyric acid (1: 1), obtained by a modified known method [Musin R.I., Lee V.A. et al. CFG, 11, p.1379-1381, 1989], 2.2 g of ASA and 0.02 ml of acetyl chloride in 40 ml of ethanol were stirred at 50 ^{° C.} for 60 minutes and at 70 ^{° C. for} 240 minutes; the solution was evacuated at 70 ^o C (-7 kPa). The resulting syrupy substance was extracted with acetone and ether, the residue was evacuated under the same conditions to form a hardened colorless foam, after grinding of which a powdery product was obtained, soluble in water, alcohol and chloroform with mp 120-125 ^o C. Yield 2.6 g (43%). Found,%: N 5.91. (C ₆ H ₉ NO ₇ ) ₅ (C ₁₈ H ₂₂ NO ₇ ) ₅ . Calculated,%: N 6.13. The content of immobilized ASA, determined by titration of a solution of SPL in alcohol with an aqueous solution of KOH, as well as by IR spectral analysis, is 39.3 wt.%.

B. 3.7 g of CPL of N-vinyl pyrrolidone with N-vinyl glycidyl-γ-aminobutyric acid (1: 1) obtained by a modified known method [Kirsh, Yu.E. Synthesis and physical. -chem. properties, M., 1998; Musin R.I. Lee V.A. et al. KhFZh, 11, p. 1379-1381, 1989], 2.2 g of ASA and 0.02 ml of acetyl chloride in 40 ml of ethanol were stirred at 50 ^° C for 60 minutes and at 70 ^° C for 240 minutes, the solution was evacuated at 70 ^° C (-7 kPa) . The resulting syrupy substance was extracted with acetone and ether, the residue was evacuated under the same conditions until a solidified colorless foam was formed, after grinding of which a powdery product was obtained, soluble in water, alcohol and chloroform, so pl. 120-125 ^o C. Yield 2.6 g (43%). Found,%: N 5.91. (C ₆ H ₉ NO) ₅ (C ₁₈ H ₂₂ NO ₇ ) ₅ . Calculated. %: N 6.13. The content of immobilized ASA, determined by titration of a solution of SPL in alcohol with an aqueous solution of KOH, as well as by IR spectral analysis, is 39.3 wt.%.

 Example 2

The solubility of IASK was determined in comparison with the substance ASA (FS 422688-94, Yilin Pharmaceutical, China). A portion of IASK 1 g, previously ground into a fine powder, was added with 10 ml of distilled water and shaken until completely dissolved at a temperature of t = 20 ^o C. After 3 minutes, no particles of the substance were detected in the solution with the naked eye. The ASA substance in an amount of 1 g, ground into a fine powder and added to a similar solvent, when shaken and at a temperature of t = 20 ^o C was completely dissolved in a volume of 1000 ml of distilled water. Thus, IASK refers according to the requirements of the State Pharmacopoeia [State Pharmacopoeia, M., 1968, p. 756] to readily soluble substances, whereas the ASA substance is poorly soluble, therefore, IASK is about 100 times superior in solubility to ASA.

 Example 3

 Experiments to study the effect of the test substance on analgesic activity were carried out against the background of intraperitoneal administration of a 3% solution of acetic acid at a dose of 300 mg / kg of animal body weight. Studies of the inventive polymer water-soluble analogue of acetylsalicylic acid and the substance of acetylsalicylic acid (ASA) (FS 422688-94, Yilin Pharmaceutical, China) were carried out according to the method described by Gatsura V.V. [book "Methods of the primary pharmacological study of biologically active substances", 1974]. All substances were administered intragastrically with a metal probe in equivalent doses. The results of the comparative evaluation are shown in table 1.

 As a result of the tests, it was found that the inventive polymer water-soluble analogue of acetylsalicylic acid in terms of ASA at a dose of 100 mg / kg of animal body weight when injected into the stomach by analgesic activity against the background of intraperitoneal injection of 3% acetic acid is 2 times higher than acetylsalicylic acid.

 Example 4

 The effect of the polymer water-soluble analogue of acetylsalicylic acid on the blood coagulation system and its morphological composition was tested on 54 white outbred male rats with a body weight of 200-250 g. Blood was studied in dynamics over the days: 1, 10, 30. Blood was taken from the tail vein ( when cutting off the tip of the tail). Registration of parameters was carried out by the device "Coagulograph portable recording type H-334." For comparison, acetylsalicylic acid (ASA) (FS 422688-94, Yilin Pharmaceutical, China) and PA were used in the same dose per ASA and in the same volume. An aqueous solution of ASA, IAAS and RA was introduced using a metal probe intragastrically daily (30 days, except Sundays).

 The results of the comparative evaluation are shown in table 2.

 An analysis of coagulograms of white rats given orally ASA and IASC allowed us to conclude that on the 10th and 30th day of the experiment, there was a change in the coagulation index (Ic), an increase in blood coagulation time after administration of ASA by 21.8%, and IASC - by 53.1% (p <0.01).

 Thus, in contrast to the effect on blood coagulation processes, IASK surpasses acetylsalicylic acid at a dose of 75 mg / day in terms of the weight of an adult 1.5-2 times. RA causes hypocoagulation (an increase in the coagulation index by 66% and a fibrinolysis index by 400%, which is a risk factor leading to bleeding).

 Example 5

 The studies were carried out on 54 white outbred male rats with a body weight of 200-250 g. During the experiment, the first group of animals daily (30 days, except Sundays) was administered orally with the help of a probe ASA preparations, the second group - RA, the third group - IASK in equivalent doses on acetylsalicylic acid (75 mg / day). Animals of the control and experimental groups were killed by decapitation (under ether anesthesia), then pathological autopsy was performed and histological examination of the stomach, liver, kidneys, heart, lungs and spleen was performed.

 At autopsy of rats from the control and experimental groups, no visible pathological changes were detected.

 However, histological examination in rats receiving ASA at a dose of 75 mg / day showed signs of increased secretion in the fundus glands of the gastric mucosa. In particular, the cytoplasm of the main (zymogenic) cells contained more secretory granules; the parietal (parietal) and additional (mucous) cells were slightly increased in volume. In the epithelium of the stomach of rats receiving IASK at a dose of 75 mg / day, there are no such changes.

 In animals of both experimental groups (treated with ASA and IASK), there was a slight increase in the volume of hepatocytes on the periphery of the hepatic lobules, an expansion of the cavity of Shumlyansky-Bowman capsules in the renal bodies, more pronounced myocardial vascularization and a slight increase in the amount of lymphoid tissue in the spleen. All these changes are physiological in nature and can be regarded as signs of some increase in the function of the organs studied.

 In animals treated with RA at a dose of 75 mg / day, signs of mucosal irritation were found in the stomachs, in particular, traces of increased mucus secretion, especially in the neck of the fundus glands, and mucus deposition on the surface of the gastric mucosa are visible. In some places, desquamation of the integumentary epithelium is observed.

 A slight degree of hydropic lesion of the tubular epithelium in the kidneys was revealed, manifested by swelling of cells, smoothing of borders, displacement of nuclei; vacuoles also appear in the cytoplasm, protein masses are visible in some tubules. In the spleens, the amount of lymphoid tissue is increased, which indicates the antigenic effect of PVP.

 Thus, IASK at a dose of 75 mg / day does not cause a damaging effect on the gastric mucosa, while ASA and RA cause irritation of the gastric mucosa and increase the secretion of the fundus glands.

Claims (2)

1. Polymer water-soluble analogue of the acid acetylsalicylic formula

where n: m = 5: 1-1: 5, with increased analgesic, anti-inflammatory, antipyretic, thrombolytic activity, reduced gastro and nephrotoxicity.  2. A method of producing a polymer of a water-soluble analogue of acetylsalicylic acid, described in paragraph 1, which consists in chemically immobilizing acetylsalicylic acid on a copolymer of N-vinylpyrrolidone with N- (glycidyl) vinyl-γ-aminobutyric acid obtained by treating polyvinylpyrrolidone with chloromethyloxy to the opening of part of the pyrrolidone cycles with the formation of the structure of N-vinyl-γ-aminobutyric acid and the formation of side chains carrying reactive glycidyl groups feasting.

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