JP2006151875A - Lipase inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an oil-soluble lipase inhibitor contributing to prevent and treat obesity caused by excessive intake of fat and diseases caused by excessive ingestion of fat and able to be added to every fat and oil. <P>SOLUTION: The lipase inhibitor comprises LUU type and UUL type triacyl glycerols (wherein L expresses a 16-22C long chain saturated fatty acid; U expresses an asymmetrical triacyl glycerol constituted of a 16-22C long chain unsaturated fatty acid) as effective ingredients and is a lipid absorption inhibitor, an anti-obesity agent and a hyperlipidemia-improving agent. Foods and medicinal agent containing the same are claimed. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a lipase inhibitor and a food containing the same. More specifically, it is highly safe to digest and absorb lipids in vivo and effectively inhibit pancreatic lipase, which is the key to obesity and hyperlipidemia, and contribute to the suppression and prevention of these diseases. The present invention relates to an oil-soluble lipase inhibitor.

In recent years, the relationship between obesity due to excessive intake of fat from the diet and lifestyle-related diseases such as diabetes, hyperlipidemia, and cardiovascular disease has been taken up and has become a problem. As a method of improving this, 1. Reduction of fat content in foods 2. Reduce the calories of the oil itself; 3. Replacement with oil substitutes 4. Promotion of metabolism, Lipase inhibitors have been proposed. 5. As for lipase inhibitors, development of drugs that suppress and prevent obesity by inhibiting the degradation of fat by pancreatic lipase has been attempted.
For example, in Patent Document 1, Red Scenic Sky, Iwabenkei, Savonsou, Bold, Paschaca, Tormentilla, El Campuri, Turmeric islet, Chuchueashi, Cat's Claw, Cinnamon, Yam, Sendangsa, Ukogi, Strawberry, Mauger, Rose, Camellia, Hypericum perforatum, Lipase inhibitors containing at least one selected from the group of plants consisting of Tochu and white tea have been proposed.

Further, in Patent Document 2, an active ingredient is an extract extracted from at least one material selected from the group consisting of yucca, ginseng, jasmine tea, yamazuko, yellow ginger tea, rooibos tea, soybean germ, ginger and tochu tea. A lipase inhibitor has been proposed.
However, the extract is almost water-soluble and cannot be mixed with fats and oils. Not on the market.
On the other hand, as an oil-soluble substance that can be dissolved in fats and oils, Patent Document 3 proposes the use of tetrahydrolipstatin as a gastrointestinal lipase inhibitor. This inhibitor is said to inactivate by covalent bonding directly with the lipase itself, its effect is quite strong, some diarrhea symptoms are also seen, food safety concerns remain, Oil-soluble lipase inhibitors that are mildly effective are desired.

JP 2002-179586 A JP 2002-275077 A US Pat. No. 4,598,089

  An object of the present invention is to provide an oil-soluble lipase inhibitor that can contribute to the prevention and treatment of obesity caused by excessive intake of fat and diseases caused by obesity, and that can be added to all fats and oils.

As a result of intensive studies to solve the above problems, the present inventors have found that an asymmetric triacylglycerol in which glycerol is composed of a long-chain saturated fatty acid at position 1 and long-chain unsaturated fatty acids at positions 2 and 3 It was discovered that asymmetric triacylglycerol composed of a long-chain saturated fatty acid at position 3 and a long-chain unsaturated fatty acid at positions 1 and 2 inhibits the activity of pancreatic lipase. It has been found that the degradation rate of the whole oil and fat is reduced by containing a small amount of in the base oil and fat.
That is, the first of the present invention is LUU type and UUL type triacylglycerol (L is a long-chain saturated fatty acid having 18 to 22 carbon atoms, U is an asymmetric type triacylglycerol composed of unsaturated fatty acid having 16 to 22 carbon atoms. ) As an active ingredient. The second is a lipid absorption inhibitor containing the asymmetric triacylglycerol described in the first as an active ingredient. The third is an anti-obesity agent containing the asymmetric triacylglycerol as described in the first item as an active ingredient. The fourth is a hyperlipidemia improving agent comprising the asymmetric triacylglycerol as described in the first item as an active ingredient. 5th is the foodstuff containing the agent in any one of 1st thru | or 4th. The sixth is a medicine containing the agent according to any one of the first to fourth.

  The LUU type and UUL type triacylglycerol of the present invention strongly inhibits lipase activity and is oil-soluble and can be added to any fats and oils for the prevention and treatment of obesity caused by excessive intake of fats and diseases caused by obesity. It is valid.

  In the present invention, the oil-soluble substance LUU type and UUL type triacylglycerol that delays hydrolysis by lipase is a saturated fatty acid whose fatty acid bonded to the 1st or 3rd position of the glycerol is a carbon number from palmitic acid having 16 carbon atoms. Long chain fatty acids are preferred, up to 22 behenic acids, and fatty acids that bind to the 2nd and 3rd positions (in the case of LUU type) or the 1st and 2nd positions (in the case of UUL type) have 16 carbon atoms. It is composed of fatty acids ranging from tooleic acid to erucic acid having 22 carbon atoms, and long-chain unsaturated fatty acids may be any of mono-unsaturated fatty acids to polyunsaturated fatty acids. From the viewpoint of safety, monounsaturation is preferable. Some examples of such LUU-type or UUL-type triacylglycerols are SOO (1st is stearic acid, 2nd and 3rd oleic acid), OOS (3rd is stearic acid, 1st and 2nd are olein) Acid), SLiLi (position 1 is stearic acid, positions 2 and 3 are linoleic acid), SLnLn (position 1 is stearic acid, positions 2 and 3 are linolenic acid), BOO (position 1 is behenic acid, positions 2 and 3 are Oleic acid), BLiO (behenic acid at the 1st position, linoleic acid at the 2nd position, oleic acid at the 3rd position), etc. Among these, BOO (behenic acid at the 1st position, oleic acid at the 2nd and 3rd positions), OOB (behenic acid at position 3 and oleic acid at positions 1 and 2) is preferred.

The LUU type and UUL type triacylglycerol oils and fats have a property that they are not easily hydrolyzed by lipase because the fatty acid bonded to the 1- or 3-position of the glycerol is a long-chain saturated fatty acid. And containing UUL type triacylglycerol in the base fat and oil in an amount of 0.5 to 30% by weight, preferably 1 to 15% by weight, and most preferably 1 to 10% by weight, so that hydrolysis by lipase is alleviated / retarded. Is.
Mild inhibitory effect is preferable as the degree of hydrolysis and delay by lipase inhibitors for fats and oils. Specifically, about 10 to 30%, preferably 10 to 20% of ingested fats and oils are ideally resistant to degradation. And
In addition, when two long-chain saturated fatty acids are introduced into the molecule, such as LLU-type and LUL-type triacylglycerol, the melting point rises and crystallization occurs near the body temperature, so that it is recognized as a lipase substrate in vivo. It becomes difficult to expect an inhibitory effect.

  This LUU-type and UUL-type triacylglycerol is generally a mixture of fats and oils containing long-chain saturated fatty acids derived from animals, plants and fish, or esters thereof, and fats and oils containing long-chain unsaturated fatty acids in any combination. Thereafter, it is obtained by transesterification by a known method using a lipase having 1,3-position specificity. Examples of vegetable oils include soybean oil, rapeseed oil, palm oil, cottonseed oil, sunflower oil, corn oil, canola oil, and animal oils include beef fat, pork fat, fish oil, and the like. Although it is possible to make non-selective transesterification, for example, using an alkaline catalyst such as sodium methylate, the regioisomers ULU, UUU, LLU, LUL, and LLL can be produced. Becomes cumbersome and not very practical. The LUU type and UUL type fats and oils thus obtained can then be concentrated to about 80% by subjecting them to ordinary processing such as fractionation.

  The LUU type and UUL type triacylglycerols of the present invention may be used as they are, or optionally blended with other base oils and fats. The blend ratio varies depending on the expected effect and the system to be used, but the base fat is not limited as long as it is an edible animal / vegetable fat.

  The LUU type and UUL type triacylglycerols of the present invention can be widely used for foods usually containing fats and oils. For example, emulsified foods such as cream, margarine, mayonnaise, dressing, dairy products, confectionery represented by chocolate, processed meat products such as bread, ham and sausage, fishery processed foods such as kamaboko and chikuwa, etc. Can be used without impairing the texture.

  The lipase inhibitor, lipid absorption inhibitor, anti-obesity agent, hyperlipidemia ameliorating agent of the present invention, and the method of administering a medicine containing these may be either oral or parenteral. In administration, the active ingredient can be mixed with a solid or liquid pharmaceutical carrier suitable for administration methods such as oral administration, rectal administration, and injection, and administered in the form of a preparation.

  EXAMPLES The present invention will be described in more detail with reference to the following examples, but the spirit of the present invention is not limited to the following examples. In the examples, “%” and “part” mean weight basis.

Production Example 1
By mixing 50 parts of high oleic sunflower oil with an iodine value of 84 and 50 parts of ethyl behenate with a purity of 95%, transesterification using a 1,3-specific lipase (Novozaymes, Lipozyme RM-IM) After the reaction oil was obtained and the esters were removed by distillation, 100 parts of hexane was further added, and crystallization and fractionation were carried out at -5 ° C to obtain 20 parts of a 70% pure LUU / UUL fraction and a 78% pure LUL fraction. 30 parts per minute were obtained.

Pharmacological test 1
The LUU / UUL fraction thus obtained was 1 part in soybean oil (LUU / UUL purity about 0.7%), 5 parts (LUU / UUL purity about 3.5%), 10 parts (LUU / UUL purity about 7%). 0.0%) The following lipase activity measurement was carried out in each of the substituted systems and the LUU / UUL fraction itself (LUU / UUL purity 70%). 80 mg of each fat and oil was sampled, phosphatidylcholine (Sigma) 80 mg, Taurocholate Na (Wako Pure Chemical Industries) 5 mg, 0.1 MTES buffer 9 ml (PH 7) containing 0.1 M NaCl were added and then ultrasonic oscillation The substrate emulsified for 1 minute was used as a substrate. 300 μl of the substrate was collected, 5 μl (5 U) of porcine pancreatic lipase (Sigma) was added, reacted at 37 ° C. for 1 hour, and then mixed with 3 ml of extraction solvent (chloroform / heptane / methanol = 49 parts / 49 parts / 2 parts). 1) and well stirred, centrifuged at 2500rpm for 5 minutes, the upper layer is removed, and copper reagent (2.98g triethanolamine, 2.42g copper nitrate, 0.48g NaOH) is dissolved in 200ml water in the lower layer. Add 1 ml of NaCl), stir for 10 minutes, centrifuge at 2500 rpm for 10 minutes, collect 1.5 ml of the upper layer, and add color reagent (0.2 g of bathocuproine, 0.1 g of butylhydroxyanisole to chloroform). The fatty acid liberated by adding 1.5 ml) was quantified by absorbance at OD480. Table 1 shows the relative activity of soybean oil itself to the system. As shown in Table 1, it was found that the activity was inhibited by about 15% in the system in which 1 part was replaced with LUU / UUL and about 25% in the system in which 5 parts were replaced.

Comparative pharmacology test 1
The LUL fraction obtained in Production Example 1 was 1 part in soybean oil (LUL purity of about 0.8%), 5 parts (LUL purity of about 3.9%), and 10 parts (LUL purity of about 7.8%). The lipase activity was measured in the same manner as in the previous test example using the substituted system and the system of the LUL fraction itself (LUL purity 78%). As shown in Table 1, even when the substitution amount of LUL was increased, no significant inhibitory effect was observed, and partial emulsion breakage due to crystallization was confirmed, and it was judged that there was no effect of inhibiting lipase hydrolysis. .

Comparative production example 1
Mixing 40 parts of tribehen with an iodine value of 1, 60 parts of ethyl oleate with a purity of 98% and 300 parts of hexane, and transesterifying using 1,3-specific lipase (Novozymes, Lipozyme RM-IM) After removing the solvent by distillation and removing the esters by distillation, 100 parts of hexane was further added, and crystallization and fractionation were carried out at a predetermined temperature to obtain 15 parts of LLU / ULL fraction having a purity of 72%. 10 parts of the 70% ULU fraction were obtained.

Comparative pharmacology test 2
The LLU / ULL fraction obtained in Comparative Production Example 1 is 1 part in soybean oil (LLU / ULL purity about 0.7%), 5 parts (LLU / ULL purity about 3.6%), 10 parts (LLU / The lipase activity was measured in the same manner as in the above-mentioned test examples in the substituted system and the LLU / ULL fraction itself (LLU / ULL purity 72%). As shown in Table 1, when the amount of substitution of LLU / ULL increased, the lipase hydrolysis inhibitory effect was somewhat recognized, but the effect as LUU / UUL was not observed.

Comparative pharmacology study 3
The ULU fraction obtained in Comparative Production Example 1 was 1 part in soybean oil (ULU purity about 0.7%), 5 parts (ULU purity about 3.5%), 10 parts (ULU purity about 7.0%). The lipase activity was measured in the same manner as in the above-mentioned test example with the substituted system and the ULU fraction itself (ULU purity 70%). As shown in Table 1, no significant lipase hydrolysis inhibitory effect was observed even when the amount of ULU substitution increased.

Digestion and absorption test in mice Using the LUU / UUL fraction obtained in Production Example 1, a digestion and absorption test in mice for about 2 months was performed. The mice used were C57BL / 6J pre-bred from 7 weeks of age for 1 week, and the diet composition was about 6 in each group with the soybean oil group as a control in the mixed feed of Table 2 with a partially improved AIN-93G composition. Breeding for 2 months, changes in body weight, feed efficiency, and body fat percentage were measured. The body fat percentage was measured using a PIXImus2 (GE Medical Systems), an X-ray bone density measuring device dedicated to experimental mice. As a result of the 56-day digestion and absorption test in mice, the group using the SUU / UUS fraction showed a decrease of about 9% in body weight and 12% in feed efficiency, and a decrease in body fat percentage of about 13%. It was observed that LUU / UUL micro-addition to obesity due to excessive intake of fat was suggested to be effective. These results are summarized in Table 3.

  According to the present invention, a lipase inhibitor that gently inhibits lipase activity, is oil-soluble, can be added to any edible oil and fat, and is effective in the prevention and treatment of obesity caused by excessive intake of fat and obesity caused by obesity And an oil / fat composition containing the same can be obtained.

Claims (6)

Lipase inhibition containing LUU-type and UUL-type triacylglycerol (L is a long-chain saturated fatty acid having 16 to 22 carbon atoms, U is an asymmetric type triacylglycerol composed of unsaturated fatty acid having 16 to 22 carbon atoms) as an active ingredient Agent. A lipid absorption inhibitor comprising the asymmetric triacylglycerol according to claim 1 as an active ingredient. An anti-obesity agent comprising the asymmetric triacylglycerol according to claim 1 as an active ingredient. The hyperlipidemia improving agent which uses the asymmetric type | mold triacylglycerol of Claim 1 as an active ingredient. A food containing the agent according to any one of claims 1 to 4. A medicament comprising the agent according to any one of claims 1 to 4.