JPH0121839B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel derivatives of chenodeoxy acids. Chenodeoxycholic acid is a type of bile acid that exists as a normal component in human bile, accounting for approximately 30% of all bile acids. In recent years, it has been discovered that this bile acid has a gallstone dissolving effect, and it has attracted attention. When chenodeoxycholic acid is orally administered to humans, it is rapidly absorbed into the body, and in the liver it conjugates with glycine or taurine to become a conjugated bile acid, which is then transferred into bile. Bile is secreted into the small intestine. The above-mentioned conjugated bile acids in bile are deconjugated to free chenodeoxycholic acid by the action of intestinal bacteria. Thereafter, chenodeoxycholic acid becomes a conjugated bile acid in the liver, and becomes a free bile acid in the intestine and circulates into the enterohepatic system, but a portion of the free bile acid becomes lithocholic acid through the action of intestinal bacteria. The amount of lithocholic acid increases with each repeat of enterohepatic circulation. Lithocholic acid is excreted into feces via the large intestine. When chenodeoxycholic acid is used as a gallstone dissolving agent, a relatively large amount is administered over a long period of time, so a relatively large amount of lithocholic acid is produced and remains in the intestines. It has been reported that lithocholic acid promotes carcinogenesis when lithocholic acid and carcinogenic substances coexist, and the present inventor believes that the above situation is unfavorable. As a result of research on bile acids, he invented sarcochenodeoxycholic acid, a new derivative of chenodeoxycholic acid. Sarcochenodeoxycholic acid is a compound of chenodeoxycholic acid and sarcosine (also known as N-methylaminoacetic acid), and its melting point is 205.5-
At 206°C (when crystallized from ethyl acetate), the specific rotation is [α] ¹⁵ _D +5.3° (methanol, C=1.0). This compound can be easily produced by a known method for synthesizing a compound having an acid amide bond from an acid and an amine. The behavior of sarcochenodeoxycholic acid in the bodies of experimental animals was found to be similar to that of chenodeoxycholic acid, except for its metabolism in the intestines. This will be explained in detail below. Intestinal absorption: 24-C ¹⁴ -sarcochenodeoxycholic acid (synthesized from 24-C ¹⁴ -chenodeoxycholic acid and sodium sarcosine) was administered into the duodenum of male Wistar rats with a bile duct cannula, and was excreted into bile within 12 hours. When the radioactivity was measured, the recovery rate was almost 100%. Also,
The majority of the radioactive compound recovered in bile was sarcochenodeoxycholic acid (determined by radio thin layer chromatography). The results of this experiment indicate that sarcochenodeoxycholic acid is rapidly absorbed into the body and transferred into bile without being broken down into free acid in the liver. Treatment with cholylglycine hydrolase: The above bile containing 24-C ¹⁴ -sarcochenodeoxycholic acid was treated with cholylglycine hydrolase (EC3.5.1.24, a conjugated bile acid hydrolase obtained from Clostridium perfuingens).
After culturing at 37°C for 2.5 hours, the reaction mixture was analyzed by radio thin layer chromatography.
Taurocholic acid spots, the main conjugated bile acid in rats, were not detected, but instead cholic acid spots were detected, while radioactivity was detected only in sarcochenodeoxycholic acid spots. This experiment showed that sarcochenodeoxycholic acid is not hydrolyzed under the conditions under which the enzyme hydrolyzes taurocholic acid. Excretion: 24-C ¹⁴ -sarcochenodeoxycholic acid was orally administered to hamsters, raised in a metabolic cage, and feces were collected to measure radioactivity in the feces. Almost 100% of the administered radioactivity was detected within 5 days.
Of these, 20% was detected from lithocholic acid. This experiment showed that sarcochenodeoxycholic acid was converted to lithocholic acid at a much lower rate than free chenodeoxycholic acid. When we compared the number of gallstones in experimental animals raised on a gallstone-forming diet and those raised on an experimental diet containing sarcochenodeoxycholic acid added to the gallstone-forming diet, we found that sarcochenodeoxycholic acid has an inhibitory effect on gallstone formation. . CD-1 mice were fed a gallstone-forming diet containing 0.5% cholesterol and 0.25% cholic acid, and a gallstone-forming diet containing 0.03% or 0.15% sarcochenodeoxycholic acid.
After being fed the supplemented experimental diet and reared for 12 weeks, the abdomen was opened to examine the formation of gallstones in the gallbladder. The number of animals forming gallstones is shown in Table 1.

[Table] This experimental result indicates that sarcochenodeoxycholic acid has an effect of inhibiting gallstone formation, and it is assumed that sarcochenodeoxycholic acid has a gallstone dissolving effect. The acute toxicity test was conducted as follows. DDY
body weight Kg in the stomach of strain mice (average weight 22.7±1.0)
Amounts of sarcochenodeoxycholic acid corresponding to 2.44 g, 3.05 g, 3.81 g, and 4.76 g per sample were dissolved in 1.5 ml of distilled water containing 1.1 equivalents of sodium bicarbonate and administered by force. As a control, only an aqueous sodium bicarbonate solution was administered in the same manner, and the survival rate after 72 hours was examined (3 to 5 animals per group). The results are shown in Table 2.

[Table] Based on these results, the LD ₅₀ of sarcochenodeoxycholic acid was determined to be 3.75 g/Kg body weight using the up-and-down method. By the way, chenodeoxycholic acid
LD ₅₀ is 3.7g/Kg body weight. This result indicates that the acute toxicity of sarcochenodeoxycholic acid is comparable to that of chenodeoxycholic acid.
The following formula was used to calculate LD ₅₀ . LD ₅₀ = 10g ^-1 [A ₀ +b ( ₃ 〓 ⁱ⁼⁰ i・Ni/N-1/2)] Example 1 Tri-n-butylamine 8 in 150ml dioxane
20 g of chenodeoxycholic acid was added and dissolved in this solution, cooled to 10° C., 4.6 ml of ethyl chlorocarbonate was added, and the mixture was stirred for 5 minutes. A solution of 6.1 g of sarcosine dissolved in 1N sodium hydroxide solution was added to this solution, stirred for 2 hours, water was added, and the mixture was concentrated under reduced pressure to obtain a syrup. When this syrup was dissolved in water and hydrochloric acid was added, a viscous precipitate precipitated out. This was crystallized from ethyl acetate to obtain 13.5 g of sarcochenodeoxycholic acid. Yield 57
%, melting point 205.5-206.0°C, specific rotation [α] ¹⁵ _D +5.3° (methanol, C=1.0). Elemental analysis values (as C ₂₇ H ₄₅ NO ₅ ) Theoretical values C 69.9%, H 9.7%, N 3.0% Experimental values C 70.0%, H 9.6%, N 2.9%

Claims (1)

[Claims] 1 formula Sarcochenodeoxycholic acid represented by.