KR100396113B1 - Method for purifying tauroursodeoxycholic acid - Google Patents

Method for purifying tauroursodeoxycholic acid Download PDF

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KR100396113B1
KR100396113B1 KR1019960045669A KR19960045669A KR100396113B1 KR 100396113 B1 KR100396113 B1 KR 100396113B1 KR 1019960045669 A KR1019960045669 A KR 1019960045669A KR 19960045669 A KR19960045669 A KR 19960045669A KR 100396113 B1 KR100396113 B1 KR 100396113B1
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acid
bile acid
acidic
xad
bile
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KR19980027016A (en
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김덕규
안상용
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동아제약 주식회사
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07JSTEROIDS
    • C07J41/00Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0038Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 with an androstane skeleton, including 18- or 19-substituted derivatives, 18-nor derivatives and also derivatives where position 17-beta is substituted by a carbon atom not directly bonded to a further carbon atom and not being part of an amide group
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07JSTEROIDS
    • C07J41/00Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0055Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives
    • C07J41/0061Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives one of the carbon atoms being part of an amide group

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Abstract

PURPOSE: A method for purifying tauroursodeoxycholic acid and a salt thereof is provided to obtain high-purity, high-yield tauroursodeoxycholic acid, which is therapeutically useful for treatment of gallstones, indigestion due to abnormal secretion of bile, or phlegm. CONSTITUTION: The method comprises: preparing a crude bile acid and dissolving or suspending the crude bile acid in an organic solvent; treating the resultant solution or suspension with an acidic or alkali agent to make an aqueous solution; saturating the aqueous solution with inorganic salts under the acidic or alkali conditions, followed by stirring for phase separation; and performing purification by repeatedly extracting and separating an organic layer or by using a non-ionic amber light XAD polymer adsorbent.

Description

타우로우르소데옥시콜린산의 정제방법Purification method of tauroursodeoxycholic acid

본 발명은 의약품으로 중요한 생리물질로서 체내기관이나 담에 널리 분포되어 있으며, 약학적으로 콜레스테롤성 담석의 용해제, 답즙분비이상으로 인한 소화불랑, 이담제 등 치료학적으로 유용한 타우로우르소데옥시콜린산을 얻기 위한 정제방법에 관한 것이다.The present invention is widely applied to medicines as important physiological substances and is widely distributed in body organs and pharmacologically. It is useful as a solubilization agent for cholesterol gallstones, digestive burn due to abnormality of secretion of bile, pharmacologically useful taurolosideoxycholic acid And a method for purifying the same.

일반적으로 타우로우루소데옥시콜린산은 우르소데옥시콜린산을 원료물질로하여 용매중에서 염기하 산무수물 등 산활성체를 제조한 후, 타우린의 염기염이나 알카리금속염으로 용해하여 축합시키는 일련의 공정으로 제조시, 공정상 어쩔수 없이 생성수반되는 여러 가지의 부생성물 및 원료, 즉 우르소데옥시콜린산, 타우린, 유기염, 무기염 등의 잔류로 물리적 화학적 방법으로 효과적으로 분리정제하기에는, 이들이 수용성이고 용해성이 유사하여 통상의 방법으로는 상당한 곤란이 따른다.In general, taururousodeoxycholic acid is a series of processes in which an acid active substance such as a base acid anhydride is prepared in a solvent using ursodeoxycholic acid as a raw material, and then dissolved and dissolved in a base salt or an alkali metal salt of taurine In order to efficiently separate and purify by various physical and chemical methods residuals of various byproducts and raw materials, such as ursodeoxycholic acid, taurine, organic salts, inorganic salts and the like, which are inevitably produced during the production process, There is a considerable difficulty in the conventional method.

이 산의 분리정제에 대한 종래의 분리정제방법을 대별하면A conventional separation and purification method for separation and purification of the acid

(1) 직접 정제법(일본화학잡지 79권, p605, 1958년, 한국공개특허 제 89-6164호,(1) direct purification method (Japanese Chemical Journal, Vol. 79, p605, 1958, Korean Patent Publication No. 89-6164,

한국특허공고 제 85-1571호)Korean Patent Publication No. 85-1571)

(2) 실리카 분리법(벨기에특허 제 901069호, 유럽특허 제 272462호)(2) Separation of silica (Belgian Patent No. 901069, European Patent No. 272462)

(3) 이온수지에 의한 정제법(유럽특허 제 400695호)(3) Purification by ionic resin (European Patent No. 400695)

(4) 전기투석법(일본공개특허 제 91-27196호) 등으로 현재 알려져 있다.(4) an electrodialysis method (Japanese Patent Laid-Open No. 91-27196).

또한 이 산의 제조는 공지방법(일본화학잡지, 79권, p605, 1958년; 일본화학잡지 76권, p505, 1995년; Acta chemical scandinavia, 7권, p1126, 1953년; Arkiv. Kemi., 8권, p331, 1955년; 벨기에특허 제 901069호 등)으로 합성이 가능하며, 본 발명자들은 종래의 제조방법을 개량하였다.The production of this acid was carried out in accordance with a known method (Japanese Chemical Journal, Vol. 79, p605, 1958, Japanese Chemical Journal, Vol. 76, p505, 1995; Acta chemical scandinavia, Vol. 7, p1126, 1953; Arkiv. Kemi., 8 , P. 331, 1955, Belgian Patent No. 901069, etc.), and the present inventors have improved the conventional production method.

한편, 종래정제 방법중 (1)의 방법 즉, 제조에서 얻어진 조결정을 에탄올이나 톨루엔 등 가용용매에 용해후 수회에 걸쳐 반복정제한 것을 대용매(Anti Solvent)로 단리하는 것으로 되어 있으나, 단순한 용해도의 차이를 이용하여 혼재하는 유무기염 및 타우로우르소데옥시콜린산유도체등으로부터 고순도의 산을 정제하여 고수율로 얻을 수 없다. (2)의 방법은 공업적으로 반복 생산하는데 있어서 회수재생 또는 재사용의 문제 및 저수율 등의 문제점이 있으며, (3)의 방법은 생성되는 부산물이 양이온성 물질 및 음이온성 물질이 동시에 혼재하기 때문에 양이온 수지와 음이온 수지를 사용함에 따라, 분리 공정조작이 많아지고 또한 저수율이다. (4)의 방법은 이온성투석막을 이용한 막분리로, 고순도로 기재되어 있으나, 이온성 분리물질과 목적산의 분자량의 차가 적어 분리조작시 목적산도 같이 투석되고 운전시 막유지가 번거로운 점 등 바람직한 방법이라고는 할 수 없다.On the other hand, the method of (1) of the conventional purification method, that is, the crude crystals obtained in the preparation are dissolved in a solvent such as ethanol or toluene and then repeatedly refined over several times to isolate the crude crystals with an antisolvent. The high purity acid can not be obtained in high yield from the mixed organic or inorganic salt and the tauroorosdeoxycholic acid derivative or the like. The method (2) has problems such as recovery / reuse and recovery in the industrial repetitive production. The method (3) has problems in that the produced by-products contain both cationic and anionic substances simultaneously, With the use of resin and anion resin, the operation of the separation step is increased and the water amount is also low. (4) is a membrane separation using an ionic dialysis membrane. However, since the ionic separation material has a small molecular weight difference between the ionic separation material and the target acid, the desired acid is dialyzed during the separation operation, It is not a method.

본 발명자들은 상기의 공지기술을 개량한 조제 담즙산의 공업적 정제방법으로 특히 타우로우르소데옥시콜린산 등 담즙산의 정제에 유효하다. 즉, 물과 분리가 되는 유기용제와 산성 또는 알칼리 수용액을 이용한 두층액 추출법에 의한 것으로처리조작이 간편하고 단시간에 분리가 가능하다. 또한 이 방법은 고순도, 고수율로 수득할 수 있어 대량공업생산의 정제법으로 우수하다.The present inventors have found that the present invention is useful for the purification of bile acid such as tauroursodeoxycholic acid, in particular, by an industrial purification method of crude bile acid improved from the above-mentioned known techniques. That is, by the two-layered liquid extraction method using an organic solvent and an acidic or alkaline aqueous solution which are separated from water, the treatment operation is simple and can be separated in a short time. In addition, this method can be obtained at a high purity and a high yield, and is excellent as a purification method for mass industrial production.

다음에 본 발명을 상세히 설명한다.The present invention will be described in detail below.

순도 55% - 95%의 조제담즙산을 물과 층분리가 가능한 유기용제 예를들면, 이소부탄올, 노르말부탄올, 이소프로판올, n-부탄올, t-부탄올, 테트라하이드로퓨란, 에틸헥산올, 에탄올 등의 단독 또는 혼합용액에 용해한다.Preparative bile acid having a purity of 55% to 95% is dissolved in an organic solvent capable of separating water and layer, for example, an organic solvent such as isobutanol, normal butanol, isopropanol, n-butanol, t-butanol, tetrahydrofuran, ethylhexanol, Or a mixed solution.

바람직하게는 이소부탄올, 이소프로판올, t-부탄올, 에틸헥산올, 2-부탄올 또는 테트라하이드로퓨란이며, 필요시 물에 불용인 헥산, 에틸아세테이트, 벤젠 등을 혼합하는 것도 좋다. 담즙산 무게당 5-20배 상기 유기용제의 사용이 다음 처리공정에 유리하다. 동량의 물을 가해 잠시 교반하고, 조제 담즙산 각 몰에 대해 산성제 0.1당량-0.5당량의 산성수용액을 가해 비교적 강한 산수용액으로 조정해 주거나, 조제담즙산 각 몰에 대해 알카리제 0.1 - 0.65당량의 알카리수용액을 가해 비교적 강한 염기성용액으로 조정해 준다. 이때 바람직하기는 산 또는 알카리의 사용량은 용액 pH에 좌우되며 이는 담즙산과 부생성물의 추출제거에 유리하다. 이 용액에 수층을 포화시키고자 수용성 무기염 즉 알칼리금속의 할라이드 등을 사용하며, 바람직하기로는 소디움 클로라이드, 포타시움 클로하이드이다. 이때 포화상태에 따라 유기용제에 추출되는 담즙산의 이행정도가 다르며, 용액중에 무기염이 소량 남은 상태가 바람직하였다.Preferred is isobutanol, isopropanol, t-butanol, ethylhexanol, 2-butanol or tetrahydrofuran, and if necessary, hexane, ethyl acetate, benzene or the like insoluble in water may be mixed. 5-20 times per weight of bile acid The use of the organic solvent is advantageous for the next treatment. The same amount of water is added and stirred for a while. An acidic aqueous solution of 0.1 equivalent to 0.5 equivalent of an acidic agent is added to each mole of the prepared bile acid to adjust the aqueous solution to a relatively strong acid, or 0.1 to 0.65 equivalents of an alkali Add an aqueous solution and adjust to a relatively strong basic solution. At this time, the amount of acid or alkali to be used is preferably dependent on the pH of the solution, which is advantageous for extracting and removing bile acids and by-products. In order to saturate the aqueous layer with this solution, a water-soluble inorganic salt such as a halide of an alkali metal or the like is preferably used, and sodium chloride and potassium chloride are preferably used. At this time, depending on the saturated state, the degree of migration of the bile acid extracted into the organic solvent is different, and it is preferable that a small amount of inorganic salt remains in the solution.

일련의 조작은 실온에서 행하며, 이상(二相: two phase)을 적절히 분배를 위해 15-30분 교반후, 방치하면 두 개의 층으로 분액이 되는데 이때 유기층을 분취한다. 이 조작을 조제담즙산의 순도에 따라 2-5회 또는 그 이상 행할 수 있다. 상기와 같은 조작에 의해 보다 수용성 원료물질인 우르소데옥시콜린산의 염, 타우린, 무기염 등이 수용액층으로 분배되어 제거될 수 있다. 산성제로서는 염산, 황산, 질산, 초산, 인산, 개미산 등이 가능하고 염산, 초산이 바람직하다. 알카리제로서는 유기알킬아민 즉, 트리에틸아민, 피리딘, 트리메틸아민, 디에틸아민 등이며, 무기알카리 즉, 소디움하이드록사이드, 암모니아용액, 포타시움 하이드록사이드 등으로 조절되며, 좋기로는 소디움하이드록사이드, 암모니아 용액이다.A series of operations is carried out at room temperature. After stirring for 15-30 minutes for proper distribution (two phases), the solution is separated into two layers, and the organic layer is separated. This operation may be carried out 2-5 times or more depending on the purity of the prepared bile acid. The salt, taurine, inorganic salt, etc. of ursodeoxycholic acid, which is a water-soluble raw material, can be distributed and removed to the aqueous solution layer by the above operation. Acidic agents include hydrochloric acid, sulfuric acid, nitric acid, acetic acid, phosphoric acid, and formic acid, and hydrochloric acid and acetic acid are preferred. Examples of the alkaline agent include organic alkyl amines such as triethylamine, pyridine, trimethylamine, diethylamine and the like, and they are controlled by inorganic alkalis such as sodium hydroxide, ammonia solution, potash hydroxides and the like, Side, ammonia solution.

이때, 수층의 포화소금물액의 추출 pH는 0.5이하를 보이며, 상층의 유기층을 탈수보조제로 처리후 농축하면 1차 정제된 조제 담즙산을 얻을 수 있는데, 액체 크로마토그래피법으포 95% 이상의 순도로 얻어진다. 얻어진 담즙산은 산성제를 사용할 때는 타우로우르소콜린산의 산성염을 알칼리제를 사용할 때 타우로콜린산의 알칼리염으로 수득된다.At this time, the extraction pH of the saturated saline solution of the water layer is 0.5 or less. When the organic layer of the upper layer is treated with a dehydrating auxiliary agent and then concentrated, primary purified purified bile acid can be obtained. . The obtained bile acid is obtained as an acid salt of taurocholic acid when an acid agent is used and an alkali salt of taurocholic acid when an alkali agent is used.

상기의 방법으로 얻어진 담즙산은 기존 방법에서 얻어진 공기중 급격한 인습문제, 부생물의 잔존문제 등을 해결하였고 고수율로 정제되었다. 보다 정제된 조재담즙산을 제조키 위해 흡착포리머 앰버라이트 XAD계 흡착제 즉, XAD-2, XAD-4, XAD-7, XAD-16 등을 사용할 수 있는데, 본 담즙산을 상용 앰버라이트의 조작에 따라 흡착할 수도 있고, 바로 흡착없이 용리할 수도 있다.The cholanic acid obtained by the above method solved abrupt convention problems in the air obtained by the conventional method and remained the by-products, and was purified at a high yield. XAD-2, XAD-4, XAD-7, XAD-16 and the like can be used in order to prepare purified purified bile acid. The present bile acid is adsorbed by the operation of commercially available amberlite Or may be eluted without adsorption.

비이온성 흡착제는 수용액중 또는 기타 극성 용매중에 녹아 있는 물질을 이온성에 무관하게 분자내 친수성 부위나, 소수성 부위를 극성결합(수소결합 등)의 성질을 이용하여 앰버라이트에 흡착 또는 분배시켜 목적하는 담즙산을 정제 분리할수 있다. 사용 XAD계 앰버라이트는 통상의 방법에 의거 활성화시켜 반복 재사용할 수 있다.The nonionic adsorbent can be prepared by adsorbing or distributing a substance dissolved in an aqueous solution or other polar solvent to a hydrophilic part in a molecule or a hydrophobic part in an amberite regardless of the ionic property using the property of polar bonding (such as hydrogen bonding) Can be purified and separated. The used XAD type amber light can be reused repeatedly by activating it according to the usual method.

본 발명자들은 상기의 이상액상 추출법에 의해 정제된 조제담즙산을 물에 용해후, pH를 중성으로 조정하고, XAD계 앰브라이트 재생방법에 의거 활성화된 앰브라이트에 흡착하여, 물, 메탄올, 아세톤, 에탄올, 암모니아수 또는 이들의 혼합용액으로 용리할 수 있다. 이때 앰브라이트 종류에 따라 설정조건이 다소 다를 수 있으나, 수세후, 그래디안트(농도증가) 메탄올 또는 아세톤에서 가장 바람직하다. 통상의 방법으로 분획하여 TLC로 확인하고 회수분을 모아 농축하고 동결건조, 스프레이드라이 또는 아세톤으로 처리하면 99.5% 이상의 고순도의 목적 담즙산을 얻을 수 있다. 순도는 액체크로마토법에 의해 측정하였다.After dissolving the prepared bile acid purified by the above-mentioned abnormal liquid extraction method into water, the present inventors adjusted the pH to neutral and adsorbed on the activated Ambalite according to the XAD amberlite regeneration method. Then, water, methanol, acetone, ethanol , Ammonia water, or a mixed solution thereof. At this time, the setting conditions may be slightly different depending on the type of the amber bright, but it is most preferable in the case of the gradual increase (concentration increase) methanol or acetone. The fraction is fractionated by a conventional method and confirmed by TLC. The collected fraction is collected and concentrated and lyophilized, spray-dried or treated with acetone to obtain a desired bile acid having a purity of 99.5% or more. The purity was measured by liquid chromatography.

이 조제 담즙산은 타우로우르소데옥시콜린산의 수화물로 되고 필요시 분획되어진 정제담즙산 수용액에 소디움하이드록사이드, 소디움바이카보레이트르 당량 투입, 조제담즙산의 나트륨염을 상기 방법으로 제조, 수득되며 기준에 상응한다.This prepared bile acid is prepared by the above method and is prepared by adding sodium hydroxide, sodium bicarbonate equivalent and sodium salt of the prepared bile acid to an aqueous solution of purified bile acid, which is a hydrate of taurosoldoxycholic acid, ≪ / RTI >

다음에 실시예로 본 발명을 더욱 상세히 설명하며, 이 실시예가 본 발명을 제한하는 것은 아니다.EXAMPLES The present invention will be described in further detail with reference to the following examples, which should not be construed as limiting the scope of the present invention.

실시예 1: 조담즙산의 제조Example 1: Preparation of crude bile acid

반응부에 테트라하이드로퓨란 120ml를 투입하고 실온에서 우르소데옥시콜린산 12.17g(0.031M)을 가하여 교반하여 용해한다. 같은 온도에서 트리에틸아민 5.0ml를 투입후 10분 교반하고 질소 기류하 빙냉하고, 10분 반응하여 산의 염을 제조한다. 같은 온도에서 메틸클로르포르메이트 2.59ml를 30분간 적하하면서 교반하여 산활성 에스테르를 제조한다. 무수물의 백탁물질에 타우린 4.5g을 노르말 수산화나트륨 30ml에 녹인 용액을 5℃ 빙욕하 30분간 가하고 동 온도에서 2시간, 상온에서 3시간 교반하여 반응을 종결시킨다. 잔류물에 이소프로필알콜 180ml를 투입 가온후, 냉각하여 불용물을 유거한다. 여액을 오일상이 될 때까지 농축한 잔사에 아세톤 150ml를 가해 결정화한 크루드 담즙산을 여과하고 건조하면 인습성의 백색결정 24g이 얻어진다. 순도는 63%(HPLC법)이다.120 ml of tetrahydrofuran was added to the reaction part, and 12.17 g (0.031 M) of ursodeoxycholic acid was added at room temperature and dissolved by stirring. At the same temperature, 5.0 ml of triethylamine is added, and the mixture is stirred for 10 minutes, cooled on a nitrogen stream, and reacted for 10 minutes to prepare an acid salt. 2.59 ml of methyl chloroformate was added dropwise at the same temperature for 30 minutes while stirring to prepare an acid active ester. A solution obtained by dissolving 4.5 g of taurine in an anhydrous cloudy material in 30 ml of normal sodium hydroxide is added to the mixture at 5 deg. C in an ice bath for 30 minutes, and the reaction is terminated by stirring at the same temperature for 2 hours and at room temperature for 3 hours. 180 ml of isopropyl alcohol is added to the residue, which is then cooled to remove the insolubles. The filtrate is concentrated to an oil phase. Crude bile acid crystallized by adding 150 ml of acetone to the residue is collected by filtration and dried to obtain 24 g of an intrinsic white crystal. The purity is 63% (HPLC method).

실시예 2: 담즙산의 2상액 정제Example 2: Purification of two-phase solution of bile acid

(1) 산성제처리(1) Acid treatment

실시예 1에서 제조한 담즙산 크루드상내의 잔류물에 순수 120ml를 가해 녹이고, 에틸아세테이트 70ml를 가해 상온에서 잠시 교반한 후 분리건조에서 방치, 분액하여 미반응 담즙산을 제거한다. 수층을 모아 교반하면서 2노르말염산 2.5ml를 가해 산성으로 하고, t-부탄올 130ml를 가한후 소디움클로라이드를 가해 수층을 포화시킨다. 다시 상온(30℃)에서 15분 교반후 분리하여 수용성 유기염 및 타우린을 제거한다.120 ml of pure water was added to the residue in the bile acid crud form prepared in Example 1, and 70 ml of ethyl acetate was added thereto. The mixture was stirred at room temperature for a while, and then separated and dried to separate unreacted bile acid. The aqueous layer was collected and stirred while 2.5 ml of 2N hydrochloric acid was added to make it acidic, 130 ml of t-butanol was added, and sodium chloride was added to saturate the aqueous layer. After stirring at room temperature (30 ° C) for 15 minutes again, the water-soluble organic salt and taurine are removed.

상층의 유기층을 모아 포화소금물 120ml를 가한후 2노르말 염산 2ml를 가해 교반하고 분리한다. 다시 위와 같은 방법으로 유기층을 회수하여 새로운 포화소금물의 상태에 따라 1-2회 추가 조작으로 액액분리추출을 실시한다. 이 조작은 추출회수에 따라 담즙산의 순도는 증가하며 4-5회가 바람직하다. 유기층에 활성탄을 가해 탈색하고 여과보조제 셀라이트를 깔아 여과 처리후 농축한 잔류물에 에탄올 10ml 및 아세톤 120ml를 가해 상온에서 결정화 한후 여과건조하면 백색결정으로 타우로 우르소콜린산의 염산염 16.3g(98%)을 얻는다. 순도는 95.8%(HPLC법)이며, 녹는점은 168-172℃이다.Collect the organic layer of the upper layer, add 120 ml of saturated brine, add 2 ml of 2N hydrochloric acid, and stir and separate. The organic layer is recovered in the same manner as described above, and the liquid separation and extraction is carried out by one or two additional operations depending on the condition of the new saturated salt water. This operation increases the purity of bile acid depending on the number of times of extraction, and is preferably 4-5 times. The organic layer was decolorized with activated charcoal, filtered, and concentrated. The filtrate was concentrated, and 10 ml of ethanol and 120 ml of acetone were added. The residue was crystallized at room temperature and then filtered to obtain 16.3 g of a hydrochloride of tauroolic acid %). The purity is 95.8% (HPLC method) and the melting point is 168-172 ° C.

(2) 알칼리제 처리(2) Alkali treatment

제 1공정의 산성제 처리방법과 동일하게 처리하되 2 노르말 염산 대신에 2노르말 가성소오다 4ml를 사용 처리하고 t-부탄올을 사용한 후 수층을 포화 소금물로 추출하여 결정화 한후 여과 건조하면 백색의 결정인 타우로우르소콜린산의 소디움염 15.4g(95%)을 얻는다. 순도는 95.2%(HPLC법)이며, 녹는점은 170-175℃이다.The same procedure as in the acidifying agent treatment in the first step was carried out except that 4 ml of 2N sodium hydroxide was used instead of 2N hydrochloric acid and t-butanol was used. The aqueous layer was extracted with saturated brine and then crystallized. 15.4 g (95%) of sodium salt of tauroolic acid is obtained. The purity is 95.2% (HPLC method) and the melting point is 170-175 ° C.

실시예3: 비이온성 포리머형 앰버라이트에 의한 정제Example 3: Purification by non-ionic polymeric amberlite

실시예 2의 제 2공정에서 제조된 담즙산의 잔류물에 순수 60ml에 용해후 2노르말 가성소오다 용액을 가해 용액의 pH를 중성으로 조정하여 미리 활성화 되고 정열을 완료한 앰버라이트 XAD-7 0.8리터에 가한후 천천히 흡착시킨다. 투입이 완료되면 순수 1.2리터를 넣어 서서히 용출, 미흡착물을 세척해낸다. 계속하여 메탄올 2.4리터를 가해 용해하면서 박층 크로마토그래피 제어 확인하고 분획 용출액을 감압 농축하여 오일상이 되면 에탄올 7ml에 용해하고, 아세톤 120ml를 가해 상온에서 결정화하면 목적의 백색 분말로 타우로 우르소콜린산의 2수화물 15.5g(93%)을 얻는다.To the residue of the bile acid prepared in the second step of Example 2 was dissolved in 60 ml of pure water and then 2 normal sodium sododate solution was added to adjust the pH of the solution to neutral. Amberlite XAD-7 0.8 liter And then adsorbed slowly. When the addition is complete, add 1.2 liters of pure water, slowly elute, and wash off the poor complex. Subsequently, 2.4 liters of methanol was added to dissolve and the mixture was analyzed by thin layer chromatography. The resulting mixture was concentrated under reduced pressure to give an oil phase. The residue was dissolved in 7 ml of ethanol. To the residue was added 120 ml of acetone and crystallized at room temperature to obtain the target white powder 15.5 g (93%) of dihydrate is obtained.

순도: 99.6%(HPLC법, 무수물로서)Purity: 99.6% (HPLC method, as anhydrous)

녹는점: 178-180℃, 215℃(분해)Melting point: 178-180 캜, 215 캜 (decomposition)

[α]D: +45˚(1% 메탄올)[?] D : + 45 ° (1% methanol)

실시예 4: 담즙산 나트륨염의 제조Example 4: Preparation of sodium salt of bile acid

정제한 타우로우르소데옥시콜린산 2수화물 5g을 물 10ml에 투입하고 소디움하이드로겐카보네이트 0.8g을 가해 잠시 가온하여 용해하고 나트륨염 4.77g(98%)을 얻는다.5 g of purified tauroorosdeoxycholic acid dihydrate is added to 10 ml of water and 0.8 g of sodium hydrogencarbonate is added and dissolved by heating for a while to obtain 4.77 g (98%) of a sodium salt.

순도: 99.7%(HPLC법, 무수물로서)Purity: 99.7% (HPLC method, as anhydrous)

녹는점 : 215-218℃ (분해)Melting point: 215-218 ℃ (decomposition)

[α]D: +35˚(1% 메탄올)[?] D : + 35 ° (1% methanol)

Claims (7)

합성된 조담즙산을 물과 분리가 되는 유기용제에 용해 또는 현탁시킨 용액에 산성제 또는 알카리제로 처리하여 수용액으로 한 다음, 산성 조건 또는 알카리 조건하에서 무기염으로 포화시킨후 교반하여 층분리하고, 유기층을 반복 추출분리하여 정제하거나, 비이온성 앰버라이트 XAD계 포리머 흡착제로 정제하는 방법을 특징으로 하는 담즙산, 그 산 또는 알칼리염, 또는 그수화물을 정제하는 방법.The resultant crude bile acid is dissolved or suspended in an organic solvent which is separated from water to give an aqueous solution which is then treated with an acidic or an alkaline agent and then saturated with an inorganic salt under acidic or alkaline conditions, Is purified by repeated extraction and purification, or purified by a nonionic amberlite XAD-based polymeric adsorbent. The method for purifying a bile acid, an acid or an alkali salt thereof, or a hydrate thereof. 제 1항에서, 담즙산이 타우로우르소데옥시콜린산인 것을 특징으로하는 방법.The method according to claim 1, wherein the bile acid is tauroursodeoxycholic acid. 제 1항에서, 유기용제가 이소프로판올, t-부탄올, 이소부탄올, 2-부탄올, n-부탄올, 테트라하이드로퓨란, 에탄올 또는 이들의 혼합용매에서 선택된 것을 특징으로 하는 방법.The method according to claim 1, wherein the organic solvent is selected from isopropanol, t-butanol, isobutanol, 2-butanol, n-butanol, tetrahydrofuran, ethanol or a mixed solvent thereof. 제 1항에서, 산성제로 염산, 황산, 초산, 개미산 또는 이들의 혼합산에서 선택된 것을 특징으로 하는 방법.The method according to claim 1, wherein the acidic agent is selected from hydrochloric acid, sulfuric acid, acetic acid, formic acid or a mixture thereof. 제 1항에서, 알카리제로 가성소오다, 암모니아, 수산화칼륨 또는 이들의 혼합 알카리에서 선택된 것을 특징으로 하는 방법.The method according to claim 1, wherein the alkaline agent is selected from caustic soda, ammonia, potassium hydroxide or a mixed alkali thereof. 제 1항에서, 무기염으로 알칼리금속의 할라이드 또는 이들의 혼합염에서 선택된 것을 특징으로 하는 방법.The method according to claim 1, wherein the inorganic salt is selected from halides of alkali metals or mixed salts thereof. 제 1항에서, 비이온성 앰버라이트 XAD계 폴리머로 비이온성 앰버라이트 XAD-2, XAD-4 또는 XAD-16에서 선택된 것을 특징으로 하는 방법.The method of claim 1, wherein the nonionic amberlite XAD polymer is selected from nonionic amberlite XAD-2, XAD-4 or XAD-16.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1985622A1 (en) * 2007-04-23 2008-10-29 Prodotti Chimici E Alimentari Spa Process for the preparation of tauroursodesoxycholic acid
WO2008128844A1 (en) * 2007-04-23 2008-10-30 Prodotti Chimici E Alimentari Spa Process for the preparation of tauroursodesoxycholic acid
WO2022059948A1 (en) 2020-09-18 2022-03-24 서울대학교 산학협력단 Method for mass-producing sodium taurodeoxycholate

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