RU2414234C1 - Method for preparing hypocholesterolemic and hypolipidemic agent of cranberry cake - Google Patents

Abstract

FIELD: medicine, pharmaceutics. ^ SUBSTANCE: invention refers to pharmaceutical industry, namely: to preparing a hypocholesterolemic and hypolipidemic agent. A method for making a hypocholesterolemic and hypolipidemic agent by triple extraction of cranberry cake, infusion, merge of extracts, distillation of a solvent, separation of an end product under certain conditions. ^ EFFECT: agent prepared by said method is characterised by high content of triterpenic acids, does not cause irritation. ^ 2 tbl, 2 ex

Description

The invention relates to the pharmaceutical industry, namely to obtain therapeutic agents from plant materials.

A known method of obtaining [US Pat. RF 2125459] biologically active substances (BAS) from plant materials by extraction with an organic solvent and subsequent dilution of the obtained extract with water. As a raw material, use sea buckthorn pulp, or a sea buckthorn leaf, or a mixture thereof, and 90-96% ethyl alcohol as an organic solvent. The extraction is carried out at the boiling point of the solvent. Obtained by this method, the product contains both a lipophilic fraction (fats, carotenoids and chlorophylls) and water-soluble biologically active substances. Triterpenic acids with biological activity, in the amount of biological substances in the specified patent do not receive.

The closest analogue (prototype) to the claimed invention is a method for producing funds with lipid-lowering and anti-atherosclerotic action, hepatoprotective, choleretic and antilithogenic activity [US Pat. RF 2011383]. According to this method, the raw meal of sea buckthorn berries remaining after the production of sea buckthorn oil is used. The extraction of raw materials is carried out with isopropyl alcohol, and the allocation of the sum of triterpenic acids from one stripped off concentrate is carried out by precipitation with petroleum ether and subsequent filtration of the target product.

The disadvantage of the prototype method is the low content of triterpene acids in the extract and a significant content of polar components, including strong organic acids (malic, oxalic and citric), which have an irritating effect. This is due to the natural feature of sea buckthorn fruits, as well as to the technology: when receiving a product from sea buckthorn meal using a polar extractant (isopropyl alcohol), a significant amount of polar ballast impurities, including strong acids, are extracted along with triterpene acids.

The preliminary extraction of native raw materials in order to obtain sea buckthorn oil in the prototype is carried out sequentially with vegetable oil and freon, while the lipophilic fraction is mainly extracted, while polar compounds, including aliphatic and triterpenic acids, remain in the meal.

The objective of the present invention is to provide a new method for producing funds based on a concentrate of triterpenic acids with hypocholesterolemic, lipid-lowering activity.

The problem is solved by the method in which a triterpenic acid concentrate is obtained from a meal of cranberry berries by triple extraction with methyl tert-butyl ether for a total of 15 hours, subsequent purification of the product from impurities by concentrating the extract to 1/5 of the volume and separating the precipitate of the target product.

The tool obtained by the present method is characterized by a high content of triterpenic acids in the extract and a low content of strong acids (malic, oxalic, tartaric, citric), which have an undesirable irritating effect compared to the tool obtained by the prototype method.

Cranberry meal, which is a waste of the food industry (production of cranberry juice), contains less ballast impurities, in particular strong acids, which makes it possible to obtain a substance that does not have an irritating effect.

The task of obtaining funds containing a higher concentration of triterpenic acids and not containing strong acids and ballast impurities is also solved by using methyl tert-butyl ether as an extractant and precipitating triterpenic acids from the extract concentrate. Methyl tert-butyl ether effectively removes triterpenic acids and poorly dissolves the polar components contained in cranberry meal, including strong acids. Non-polar and polar impurity compounds from cranberry extracts remain in the mother liquors, separated from sediments containing triterpene acid concentrates.

The proposed method is as follows.

Cranberry meal is extracted 3 times with methyl tert-butyl ether for a total of 15 hours, the combined extract is concentrated at 60-70 ° C to 1/5 of the original volume, after which the evaporated solution is kept and the precipitate containing triterpenic acids is separated from the mother liquor containing lipophilic and polar impurities connections. After draining the organic solvent after draining the organic solvent containing the residual amount of solvent, water is charged and, when heated to 80-90 ° C, the residual solvent is distilled off from the meal with water vapor.

The agent obtained from cranberry meal according to the proposed method has increased activity, contains more triterpene acids, does not have an irritating effect and can be used in a lower dosage.

Example 1. Description of the method of obtaining funds

80 kg of cranberry meal are poured into 400 l of methyl tert-butyl ether, insisted for 5 hours. The extraction is repeated two more times. The combined extracts are concentrated to 1/5 volume. Concentrates remaining in the cube after distillation of the extractant, incubated for 24 hours to separate the precipitation of triterpenic acids (TTK). The mother liquors containing a solution of low-polar impurities in methyl tert-butyl ether are separated by decantation, the remaining amount of mother liquors is separated by filtration. The precipitate of triterpenic acids is dried in an oven at a temperature of 40-50 ° C. The yield of 2.8-3.6 kg (35-45 g / kg of meal 3.5-4.5%) with the content of the sum of ursolic and oleanolic acids 75% (titrimetrically), acid number 1.6 mg KOH / g.

The tool obtained by the prototype method is characterized by a triterpenic acid content of 62% (titrimetrically), an acid number of 151 mg KOH / g, 1.8% yield.

The tool obtained by the present method by extraction of cranberry meal with methyl tertiary butyl ether with the precipitation of the extract concentrate, is characterized by a high content of triterpene acids, practically does not contain strong acids and is released in higher yield compared to the tool obtained by the prototype method.

Example 2. Tests of the funds obtained by the claimed method

Experimental hypercholesterolemia in white rats was caused by intraperitoneal administration of Tween-80 at the rate of 200 mg / 100 g body weight. At the same time, the test agent was administered orally at doses of 10-500 mg / kg. After 12 hours, the rats were sacrificed by decapitation and the cholesterol content was determined in the blood serum. As a comparison substance, the sea buckthorn extract obtained by the prototype method was used, the hypocholesterolemic effect of which was studied previously.

As experiments showed (Table 1), as a result of the introduction of Tween-80 in rats of the control group, the level of cholesterol in the blood compared with intact animals increased significantly (by 52%). The introduction of the test agent delayed the increase in blood cholesterol in animals under the influence of Tween-80, doses of 10 and 50, 100, 200 mg / kg caused a decrease in cholesterol of 30-38%.

The hypocholesterolemic effect of the agent obtained by the present method exceeded the effect of the sea buckthorn extract (obtained by the prototype method), which may be due to the higher content of triterpenic acids and the absence of side and irritating substances that can reduce the hypocholesterolemic effect of the agent.

From the data presented in table 1, it is seen that the agent obtained by the present method more effectively reduces cholesterol compared to the sea buckthorn agent obtained by the prototype method, in dosages of 10 mg / kg -100 mg / kg, and in dosages of 200 mg / kg - 500 mg / kg, not inferior to the remedy from sea buckthorn.

To assess the lipid-lowering effect of the agent obtained by the present method, the following biochemical parameters were determined: in the blood - the content of total lipids, β-lipoproteins and pre-β-lipoproteins. The data are presented in table 2.

From the data presented in table 2, it is seen that as a result of the use of the drug, there is an improvement in blood indices (total lipids; β-lipoproteins and pre-β-lipoproteins) compared with the control at a dosage of 10 to 500 mg / kg.

Table 1
The effect of the funds obtained by the present method, and the extract of sea buckthorn (obtained by the prototype method) on the cholesterol in the blood of experimental animals Sea buckthorn extract (prototype method) n = 6 The tool according to the claimed method n = 6 Intact animals n = 6 Control, n = 13 Tween-80 and 10 mg / kg 1.54 ± 0.063
P _to > 0.05
P _and <0.05
-17% 1.31 ± 0.084
P _to > 0.05
P _and > 0.05
-thirty% 1.22 ± 0.111 1.86 ± 0.1
91 P _and <0.05
+ 52% Tween-80 and 50 mg / kg 1.40 ± 0.020
P _to > 0.05
-27%
P _and > 0.001 1.15 ± 0.110
P _to <0.01
-38%
P _and > 0.05 Tween-80 and 100 mg / kg 1.34 ± 0.106
P _to > 0.05
-28%
P _and > 0.001 1.17 ± 0.185
P _to <0.01
-37%
P _and > 0.05 Tween-80 and 200 mg / kg 1.21 ± 0.111
P _to <0.01
-35%
P _and > 0.05 1.20 ± 0.104 P _to <0.01
-35%
P _and > 0.05 Tween-80 and 500 mg / kg 1.28 ± 0.111
P _to > 0.05
P _and > 0.001 1.2 ± 0.089
P _to <0.01
-thirty%
P _and > 0.05 Note: R _And - the level of significant difference in relation to intact values; R _To - the level of significant differences in relation to the control values; n is the number of animals in the group

table 2
The influence of the funds obtained by the present method on lipid metabolism in the blood against the background of twin hyperlipidemia Groups of animals Blood ol, g / l β- and pre-β-LP blood, g / l Intact, n = 6 1.31 ± 0.288 0.73 + 0.129 Control, n = 13 3.29 ± 0.247
P _and <0.001
+ 151% 3.15 + 0.385
P _and <0.001
+ 332% TK, 10 mg / kg, n = 6 2.92 + 0.199
P _to > 0.05
P _and <0.001
+ 123% 2.49 + 0.304
P _to > 0.05
P _and <0.001
+ 241% TK, 50 mg / kg, n = 6 2.33 + 0.188
P _to <0.05
-29%
P _and <0.001 + 123% 2.15 + 0.401
P, <> 0.05
P _and <0.001
+ 195% TK, 100 mg / kg, n = 5 2.08 ± 0.139
P _to <0.05 - 2.22 + 0.232
P, <> 0.05 TK, 200 mg / kg, n = 6 2.10 ± 0.123
P _to <0.05 - 1.84 + 0.142
P _to <0.05- TK, 500 mg / kg, n = 6 1.79 + 0.007
P _to <0.05 - 1.70 + 0.220
P _to <0.05 -

Claims (1)

A method of obtaining funds with hypocholesterolemic and lipid-lowering effects by triple extraction of berry meal, insisting, combining extracts, distilling off the solvent, separating the target product, characterized in that cranberry berry meal is used as a meal of berries, extraction is carried out at a ratio of cranberry berry meal: methyl tertbutyl ether 1: 5 for 15 hours, the combined extracts are concentrated to 1/5 of the volume, the resulting concentrates are kept for 24 hours to separate the precipitate of triterpenic acids, wasp approx dried at a temperature of 40-50 ° C, wherein the product comprises at least 75 wt.% of triterpenic acids.