Samandarov N.Yu.
SYNTHESIS AND BIOCHEMICAL STUDY OF CHOLELYTOLYTIC, HEPATOPROTECTIVE PREPARATIONS "URSOSLITA" *
Аннотация:
in this paper author makes synthesis and biochemical study of cholelytolytic, hepatoprotective preparations "ursoslita"
Ключевые слова:
synthesis, biochemical, cholelytolytic
УДК 54
Samandarov N.Yu.
Scientific Research Institute
"Institute of Gastroenterology of the Republic of Tajikistan"
Tajik National University State Institution
(Dushanbe, Tajikistan)
SYNTHESIS AND BIOCHEMICAL STUDY OF CHOLELYTOLYTIC,
HEPATOPROTECTIVE PREPARATIONS "URSOSLITA"
Abstract: in this paper author makes synthesis and biochemical study of cholelytolytic, hepatoprotective preparations "ursoslita".
Keyword: synthesis, biochemical, cholelytolytic.
One of the important achievements of modern medicine is the establishment in the early 70s of the fact that chenodeoxycholic acid causes a pronounced decrease in the lithogenicity of bile in patients with cholelithiasis. This was the beginning of the chemical dissolution of cholesterol stones in the gallbladder.
The incoming chenodeoxycholic acid reduces the secretion of cholesterol in bile, due to which gallbladder bile loses its lithogenicity. It has been shown that the activity of HMG-CoA reductase in patients with cholesterol gallstones increases.
Reducing the concentration of cholesterol in bile, according to H.H. Mansurov [1-3].
During the course of treatment with CDCA, every 2 weeks, the lithogenicity of bile was monitored and the activity of enzymes of the transamination process was determined.
The structure of the liver after 6-8 months of treatment with chenodeoxycholic acid did not show significant changes, while a slightly greater accumulation of fat in hepatocytes and an increase in the content of litofucin could be noted. Under the wobbling of chenotherapy, a natural dynamics was observed on the part of bile chemistry. The content of total bilirubin gradually decreased, the amount of total cholesterol decreased markedly; at the same time, the amount of total bile acids increased sharply, mainly due to chenodeoxycholic acid. The level of phospholipids in the course of chenotherapy significantly increased, favoring the process of dissolution of cholesterol gallstones. As a result, the cholato-cholesterol ratios were normalized.
Ursofol hydrophilic non-toxic tertiary bile acid formed under the action of bacterial enzymes from 7-keto-lithocholic acid, which entered the liver from the small intestine, was of the greatest importance in the treatment of liver diseases from drugs of different groups.
Ursadeoxycholic acid appears to be the most effective agent used for the pathogenetic treatment of primary biliary cirrhosis and primary sclerosing cholangitis. The effect of ursodeoxycholic acid on the course of primary biliary cirrhosis has been evaluated in a number of detailed randomized studies, in most of which, the dose of the drug was 13-15 mg per 1 kg of body weight per day.
According to most researchers, ursodeoxycholic acid is most effective at stage III of primary biliary cirrhosis and ineffective at the last –IV stage of the disease.
To date, none of the hepatoprotectors used in medical practice fully satisfy these requirements, although in recent years the arsenal of modern hepatoprotective agents has expanded due to the appearance of both synthetic drugs and new natural remedies.
Depending on the chemical structure and origin, there are several groups of hepatoprotectors:
These include silymarin, liv-52, essential phospholipids, amino acids, vitamins C, E, and finally drugs of different groups, for example, UDCA.
In the treatment of liver diseases, of all existing drugs, ursadeoxycholic acid is of the greatest importance. By its properties, it is a non-toxic tertiary bile acid formed under the action of bacterial enzymes from 7α-keto-3α-hydroxy-5β-cholanic acid, which enters the liver from the small intestine. When using the drug, the enterohepatic circulation of hydrophobic cholanic acids decreases, their toxic effect on the membranes of hepatocytes and on the epithelium of the bile ducts is prevented.
Thanks to the conducted experimental and clinical experiments, which indicate that ursadeoxycholic acid has hepatoprotective, anticholestatic, immunomodulating, hypocholesterolemic, litholytic effects.
Ursadeoxycholic acid has a therapeutic effect in non-alcoholic steatohepatitis, primary biliary cirrhosis. However, it reduces the serum bilirubin content by 25%, the activity of alanine aminotransferase (ALT) by 35%, aspartate aminotransferase (AST) by 33%, alkaline phosphatase by 40% and γ-glutamyl transpeptidase by 50%.
In the liver and intestinal mucosa, almost the only way of metabolism of ursadeoxycholic acid occurs conjugation, that is, it forms esters with taurine and glycine. The main way of its transformation by the intestinal microflora is the formation of 7-keto-3α-hydroxy 5β-cholanic acid or 3α-hydroxy-5β-cholanic acid.
Lithocholic acid contributes to cholestasis, which has an adverse effect on the liver up to death due to liver failure in patients with sulfonation disorder. When using the drug, the enterohepatic circulation of hydrophobic cholanic acids decreases, their toxic effect on the membranes of hepatocytes and on the epithelium of the bile ducts is prevented.
Thanks to the conducted experimental and clinical experiments, which indicate that ursadeoxycholic acid has hepatoprotective, anticholestatic, immunomodulating, hypocholesterolemic, litholytic effects.
Ursadeoxycholic acid has a therapeutic effect in non-alcoholic steatohepatitis, primary biliary cirrhosis. However, it reduces the serum bilirubin content by 25%, the activity of alanine aminotransferase (ALT) by 35%, aspartate aminotransferase (AST) by 33%, alkaline phosphatase by 40% and γ-glutamyl transpeptidase by 50%.
In the liver and intestinal mucosa, almost the only way of metabolism of ursadeoxycholic acid occurs conjugation, that is, it forms esters with taurine and glycine. The main way of its transformation by the intestinal microflora is the formation of 7-keto-3α-hydroxy 5β-cholanic acid or 3α-hydroxy-5β-cholanic acid.
Lithocholic acid contributes to cholestasis, which has an adverse effect on the liver up to death due to liver failure in patients with sulfonation disorder. Lithocholic acid causes DNA breaks, being a strong co-mutagen, promoting cell transformation, leads to segmental damage to the bile ducts, liver failure. As shown by gas chromatographic data, normally up to 44% of ursadeoxycholic acid is converted into 3α-hydroxy-5β-cholanic acid within 2 hours of incubation in vitro and up to 100% in vivo for 12 to 24 hours. lithocholic in experiments of patients receiving ursadeoxycholic acid.
In clinical studies of ursodeoxycholic acid, as a rule, the level of the formed lithocholic acid was not determined, although it is known that the oxidation or reduction of this drug occurs at the 7α-position of the steroid molecule.
It should be noted that in all cases of the use of ursadeoxycholic acid for the treatment of patients with liver pathology, the level of ursadeoxycholic acid was not determined (4).
The search for literature data shows that until recently there is no information on gas chromatographic analysis of the content of bile acids in the blood serum in patients with steatosis and steatohepatitis, as well as in cases of using any drugs in the course of therapy.
Therefore, the use of more sensitive methods for determining the content of bile acids is an urgent problem. It would be interesting to consider the content of bile acids in the blood serum in patients taking ursadeoxycholic acid using gas-liquid chromatography.
When assessing the content of bile acids in the blood serum of patients with fatty liver disease: it was found that in fatty liver disease in the case of steatosis, the content of bile acids is: cholic - 0.42 ± 0.08 mg / ml; chenodeoxycholic - 0.092 ± 0.001 mg / ml; dehydrocholic - 0.013 mg / ml; deoxycholic - 0.037 ± 0.007 mg / ml, and lithocholic acid - 0.016 ± 0.03 mg / ml.
However, in the case of steatohepatitis, a noticeable increase in the concentration of chenodeoxy-0.11 ± 0.022 mg / ml was also noted; cholic-1.17 mg / ml; deoxycholic - 0.041 ± 0.003 mg / ml; dehydrocholic acid - 0.15 ± 0.004 mg / ml and lithocholic acid - 0.037 mg / ml.
The data obtained indicate that the total content of bile acids significantly increased in patients with steatosis and steatohepatitis.
The result of gas chromatographic analysis of bile acids in patients with steatohepatitis after taking ursadeoxycholic acid at a dose of 15 mg / kg per body weight for 2 months takes the following form.
The data obtained after therapy show that the concentration of cholic is 0.0097 mg / ml; chenodeoxycholic - 0.0081 mg / ml; g deoxycholic - 0.0070mg / ml; dehydrocholeau - 0.0029 mg / ml; and lithocholic acid - 0.014 mg / ml, respectively.
As can be seen from the above results of gas chromatographic analysis of bile acids, in patients with steatohepatitis before and after treatment with ursadeoxycholic acid for 2 months, strictly according to certain indications, there is a significant normalization of bile acids, except for lithocholic acid.
The increase in the concentration of lithocholanic acid in the blood serum of patients with steatohepatitis was 0.018 mg / ml of the total bile acids, which indicates that part of the ursofalk in the body is metabolized with the formation of lithocholic acid causing DNA breaks.
Thus, the obtained gas chromatographic results confirm the data on some toxic factors of ursadeoxycholic acid.
In recent days, the number of patients with various hepatobiliary pathologies in the world has exceeded 2 billion people. If we take into account the CIS countries, then annually up to 1 million people are registered suffering from this or that liver disease.
Considering all this, it should be noted that the significant spread of diseases of the liver, gallbladder, obesity and the difficulty of recognizing them, as well as more effective treatment, prompts us to look for more suitable new drugs, based on certain steroids such as bile acids.
At present, the search for new litholytic, hypocholesterolemic, hypolipidemic and hepatoprotective agents remains an unresolved issue in the development of bioorganic chemistry and biochemistry.
A rational chemical, biochemical and pharmacological approach to the production of certain substances is based on an assessment of the possible mechanism of their biotransformation or on structural analogs with known pharmacologically active compounds. All newly obtained drugs are tested in biochemical or biological laboratories, where their effect in vitro or in vivo on infected experimental animals is studied.
In order to create litholytic, hypocholesterolemic and hepatoprotective agents, attempts were made to establish a relationship between the chemical structure and biological activity. This prompted us to carry out a targeted synthesis of pharmacologically active agents based on certain steroids such as bile acids, which have increased selectivity, efficacy, stability and duration of action.
The solution to the set task of our research also included the production of sodium salt of 3α, 7β-dihydroxy-5β-cholanic acid in order to synthesize substances with cholelitic, hypocholesterolemic, hypolipidemic and hepatoprotective properties.
The neutralization of 3α, 7β-dihydroxy-5β-cholanic acid was carried out by its interaction with sodium hydroxide in a dioxane solution.
In this regard, we have carried out the synthesis of propane-1,2-diol ester of 3α, 7β-dihydroxy-5β-cholanic acid (III) by the interaction of the sodium salt of the corresponding cholanic acid (II) with glycerol α-monochlorohydrin.
It should be noted, after cleaning and establishing the structure of ursoslite by physicochemical methods, that the study was directed, mainly from the position of studying choleretic, hypolipidemic, hypocholesterolemic, hepatoprotective and litholytic properties, this explains the choice of the direction of work.
"Ursoslit" in accordance with this work is a new, undescribed in the literature derivative of "Ursofalk" -а propane-1,2-diol ester 3α, 7β-dihydroxy-5β-cholanic acid, belonging to the group of steroids of the type of bile acids, and exhibiting the best cholelitholytic, choleretic, hepatoprotective, hypocholesterolemic and hypolipidemic actions. These qualities are manifested due to the ester group.
In order to study the cholelitic and hypocholesterolemic properties of Ursoslit, we conducted experiments on 20 hamsters of both sexes, weighing 55-70 g. ... 2-untreated hamsters who received CGFD for 6 months (cholelitogenic-hyperlipidemic diet), experienced, who, along with CGFD for 6 months, daily intragastric administration of "Ursoslit" at a dose of 50 mg / kg body weight, 4 animals, who received HGLD and "Ursofalk" at a dose of 50 mg / kg body weight according to the same scheme.
The effectiveness of the studied drugs was judged by:
In the group of animals that received daily for 6 months, along with dry CGLD, the study "Ursoslit" at a dose of 50 mg / kg of body weight was found only in the gallbladder of 1 animal out of 5, and 1 stone with a size of 2.0 mm was found. Sand and small stones were absent in all cases.
In animals that received the same scheme Ursofalk at a dose of 50 mg / kg body weight, gallbladder calculi were found in 3 out of 5 cases, which was 60%.
Thus, "Ursoslit" at a dose of 50 mg / kg of body weight, on average, 3-4 times more actively prevented the occurrence of cholelithiasis in experimental chomiyaks than "Ursofalk" at the same dose.
Biochemical studies in hamsters that had been fed a cholelitogenic diet for 6 months showed that they sharply disrupt the chemistry of bile in the direction of increasing its lithogenicity. There was also a statistically significant increase in the concentration of cholesterol and especially bilirubin in bile (P. <0.001).
Thus, "Ursoslit" shows an active therapeutic effect in experimental cholelithiasis and is not inferior in degree of effectiveness, but on the contrary surpasses the activity of "Ursofalk" -a. To confirm the above and for the purpose of conducting a biochemical study to study the litholytic properties of "Ursoslit" -а, calculi, it was necessary to clarify the content.
Biochemical synthesis of bile acids in the body of animals and humans is carried out in the smooth endoplasmic reticulum of hepatocytes, mainly from newly synthesized cholesterol. In this regard, the quantitative determination of bile acids in bile plays an important role in making an accurate diagnosis, as well as effective treatment.
On the basis of gas chromatographic data, a graph of the content of bile acids was plotted depending on the intake of cholelitogenic-hyperlipidemic diet (HGLD) by hamsters for 6 months, in the process of treatment with Ursoslit and Ursofalk.
From the studies carried out, the following conclusion can be drawn: "Ursoslit" exhibits pronounced cholelitic, hypocholesterolemic, hypolipidemic and hepatoprotective actions, increases the content of total bile acids and phospholipids, reduces the high lithogenicity of bile.
"Ursoslit" exhibits an active therapeutic effect in experimental cholelithiasis and surpasses the activity of ursodeoxycholic acid in terms of efficiency. This is manifested:
"Ursoslit" refers to a true cholelitic, litholytic, hepatoprotective agent, and the studies carried out make it possible to recommend it for the treatment and prevention of gallstone and other liver diseases.
Gas chromatographic analysis of higher fatty acids in bile in experimental hamsters in the case of CGLD + Ursoslit always shows an increase in unsaturated fatty acids, which can lead to the development of positive dynamics from the effective action of Ursoslit.
It was shown that during gas chromatographic analysis of higher fatty acids in bile in hamsters, with experimental cholelithiasis under the influence of Ursoslit, the content of unsaturated higher fatty acids increased significantly: for example, oleic - (23.89 ± 0.73%), linoleic - (10.27 ± 0.16%), linolenic acid (5.26 ± 0.11%) and arachidonic acid (15.25 ± 0.57%) compared with bile in hamsters who received dry CGLD.
Gas chromatographic methods have established the effect of ursodeoxycholic acid on the nature of changes in the content of cholesterol and bile acids in fatty liver disease. Under the action of ursodeoxycholic acid in bile and blood serum, the amount of bile acids, especially chenodeoxycholic acid, increases. Ursodeoxycholic acid also stimulates the increase in unsaturated fatty acids and suppresses unsaturated fatty acids, which also reduces the degree of hepatic acid steatosis.
The results of gas chromatographic analysis of cholanic acids, in patients with steatohepatitis before and after treatment with ursodeoxycholic acid for 2 months, strictly according to certain indications, there is a significant normalization of bile acids, except for lithocholic acid.
The increase in the concentration of lithocholic acid in the blood serum of patients with steatohepatitis was 0.018 mg / ml of the total amount of bile acids, which indicates that part of the ursodeoxycholic acid in the body is metabolized to form lithocholic acid causing DNA breaks.
The obtained gas chromatographic results confirm some toxic factors of ursodeoxycholic acid.
Obtaining a new analogue of bile acids (Kaholit) and an isolated plant agent "Fitasuman" exhibiting antimicrobial activity against field strains, staphylococcus, nocardia, corynobacteria, pastearella and is not inferior to synthetic drugs of a similar purpose.
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Номер журнала Вестник науки №10 (55) том 2
Ссылка для цитирования:
Samandarov N.Yu. SYNTHESIS AND BIOCHEMICAL STUDY OF CHOLELYTOLYTIC, HEPATOPROTECTIVE PREPARATIONS "URSOSLITA" // Вестник науки №10 (55) том 2. С. 106 - 116. 2022 г. ISSN 2712-8849 // Электронный ресурс: https://www.вестник-науки.рф/article/6267 (дата обращения: 20.04.2024 г.)
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