KR100355533B1 - Process for the stereoselective synthesis of nucleoside analogues - Google Patents

Abstract

본 발명은 2R-히드록시메틸-5S-(5'-플루오로시토신 -1'-일)-1,3-옥사티올란의 제조방법에 관한 것으로, a)(2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산 (1'R,2'S,5'R)멘틸에스테르에 5-플루오로우라실을 커플링시켜 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조한 다음, b)상기 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란 -2R-카르복실래이트를 활성화시킨 후 아민화하여 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조하고, c)상기 (1'R, 2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 환원시키는 것으로 구성되는 것을 특징으로 한다.The present invention relates to a method for preparing 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane, comprising: a) (2R, 5R) -5-hydroxy -[1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) by coupling 5-fluorouracil to menthyl ester (1'R, 2'S, 5'R)- Menthyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate was prepared, and b) the above (1'R, 2'S, 5 ' R) -mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate is activated and then aminated to give (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate, and c) the (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate. .

Description

뉴클레오시드 유사체의 입체선택적 합성방법{Process for the stereoselective synthesis of nucleoside analogues}Process for the stereoselective synthesis of nucleoside analogues

본 발명은 뉴클레오시드 유사체의 입체 선택적 합성 벙법에 관한 것으로, 특히 2R-히드록시메틸-5S-(5'-플루오로시토신-1'-일)-1,3-옥사티올란((-)-FTC)의 합성방법에 관한 것이다.The present invention relates to stereoselective synthesis of nucleoside analogs, in particular 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane ((-)- To a method of synthesizing FTC).

인체 면역 결핍 바이러스와 B형 간염 바이러스에 대해 항바이러스 활성을 갖는 뉴클레오시드 유사체들이 알려져 있다. 이러한 뉴클레오시드 유사체들은 대부분 두개 이상의 카이랄 센터를 가지고 있고, 크게는 시스 및 트랜스 광학 이성질체 각각의 쌍을 가지고 있다. 이 가운데 한가지 시스 이성질체 만이 일반적으로 생물학적 활성과 낮은 세포독성을 나타낸다. 따라서, 이러한 한가지 시스-뉴클레오시드의 입체 선택적 합성 기술이 매우 중요하다.Nucleoside analogs with antiviral activity against human immunodeficiency virus and hepatitis B virus are known. These nucleoside analogues mostly have two or more chiral centers, and largely have pairs of cis and trans optical isomers, respectively. Only one cis isomer generally shows biological activity and low cytotoxicity. Therefore, the technique of stereoselective synthesis of one such cis-nucleoside is very important.

뉴클레오시드 유사체의 제조방법으로는 국제특허공개공보 WO92/20669, 및 WO95/29176 호에 게시되어 있는 바와 같이, 루이스 산을 이용해 1.3-옥사 티올란 유도체에 퓨린 또는 피리미딘 염기를 글리코실화하는 방법이 있다. 그러나, 이 방법으로 제조하게 되면 시스형태와 트랜스 형태의 뉴클레오시드 유사체를 분리해야하므로 생물학적 활성이 있는 시스 형태의 수율이 낮은 단점이 있다.Methods for preparing nucleoside analogs include glycosylation of purine or pyrimidine bases on 1.3-oxa thiolane derivatives using Lewis acids, as disclosed in WO 92/20669 and WO 95/29176. have. However, this method has a disadvantage in that the yield of the cis form with biological activity is low because the nucleoside analogue of the cis form and the trans form must be separated.

국제특허공개공보 WO94/29301 호에는 비사이클릭 중간체를 이용하여 입체 선택적으로 뉴클레오시드를 합성하여 2R-히드록시메틸-5S-(5'-플루오로시토신-1'-일) -1,3-옥사티올란((-)-FTC)를 제조하는 방법이 게시되어 있다. 그러나, 상기 방법들은 모두 루이스 산을 이용해야 하는 단점이 있다.International Patent Publication No. WO94 / 29301 discloses 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3 by synthesizing nucleosides stereoselectively using bicyclic intermediates. A method for preparing oxathiolane ((-)-FTC) is disclosed. However, all of the above methods have the disadvantage of using Lewis acid.

루이스 산을 이용하지 않고 뉴클레오시드 유사체를 합성하는 방법이 WO95/29174호에 게시되어 있으나, 이 방법도 완전하게 한가지 시스 형태만의 뉴클레오시드 유사체를 합성하지는 못했다. 따라서, 동반 생성되는 광학 이성질체를 효소나 크로마토그램으로 분리해야하는 문제가 있었다.A method for synthesizing nucleoside analogs without the use of Lewis acids has been published in WO95 / 29174, but this method also did not synthesize nucleoside analogs of only one cis form. Therefore, there is a problem in that the optical isomers generated together must be separated by enzymes or chromatograms.

따라서, 본 발명의 목적은 루이스 산을 사용하지 않으면서 입체 선택적으로 뉴클레오시드 유사체를 제조하는 방법을 제공하는 것이다.It is therefore an object of the present invention to provide a method for preparing nucleoside analogs that are stereoselectively without the use of Lewis acids.

본 발명의 다른 목적은 저가의 원료를 사용하여 높은 수율로 2R-히드록시메틸-5S-(5'-플루오로시토신 -1'-일)-1,3-옥사티올란((-)-FTC)를 제조하는 방법을 제공하는 것이다.Another object of the present invention is to provide 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane ((-)-FTC) in high yield using inexpensive raw materials. It is to provide a method for producing.

상기 본 발명의 목적은 a)(2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R)멘틸에스테르에 5-플루오로우라실을 커플링시켜 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조한 다음, b)상기 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 활성화시킨 후 아민화하여 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조하고, c)상기 (1'R, 2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 환원시키는 것으로 구성되는 것을 특징으로 하는 2R-히드록시메틸-5S-(5'-플루오로시토신 -1'-일)-1,3-옥사티올란의 제조방법에 의해 달성된다.The object of the present invention is a) 5-fluoro to (2R, 5R) -5-hydroxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) menthyl ester Coupling the lauracil to (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylay B) (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R- After activating the carboxylate, it is then aminated to (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane- 2R-carboxylate, and c) (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxa To a process for producing 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane, characterized by reducing thiolane-2R-carboxylate. Is achieved.

본 발명의 2R-히드록시메틸-5S-(5'-플루오로시토신 -1'-일)-1,3-옥사티올란의 합성 방법을 다음 화학반응식에 도시하였다.The synthesis method of 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane of the present invention is shown in the following chemical scheme.

1) 타르타르산(I)에-멘톨과 p-톨루엔설폰산을 반응시켜 (1'R,2'S,5'R)-디멘틸에스테르(II)를 제조한다. 이때 용매로 공비가 가능한 혼합용매를 사용하는 것이 효과적이다. 예를 들어 톨루엔, 벤젠, 및 에틸아세테이트 중 하나와 테트라히드로 퓨란, 아세토니트릴 중 하나의 혼합용매를 사용할 수 있고, 특히 톨루엔과 테트라히드로퓨란 혼합용매를 사용하는 것이 바람직하다.1) to tartaric acid (I) (1'R, 2'S, 5'R) -dimentyl ester (II) is prepared by reacting menthol with p-toluenesulfonic acid. At this time, it is effective to use a mixed solvent which can be azeotropic as a solvent. For example, a mixed solvent of one of toluene, benzene, and ethyl acetate and one of tetrahydrofuran and acetonitrile can be used, and it is particularly preferable to use a mixed solvent of toluene and tetrahydrofuran.

2)타르타르산(1'R,2'S,5'R)-디멘틸에스테르(II)를 과요오드화나트륨 및 소듐티오설패이트를 사용하여 (1'R,2'S,5'R)-멘틸글리옥실래이트(III)로 만든다.2) Tartaric acid (1'R, 2'S, 5'R) -dimentyl ester (II) using sodium iodide and sodium thiosulfate Made with t (III).

3)(1'R,2'S,5'R)-멘틸글리콕실래이트(III)를 1,4-디티안-2,5-디올(IV)과 반응시켜 (2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R)멘틸에스테르(V)를 제조한다.3) (1'R, 2'S, 5'R) -Mentylglycolate (III) is reacted with 1,4-dithiane-2,5-diol (IV) to give (2R, 5R) -5-hydride Roxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) mentyl ester (V) is prepared.

4)(2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R)멘틸에스테르(V)에 5-플루오로우라실을 커플링시켜 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VII)를 제조한다. 이 커플링 반응에서는 약 유기산, 예를 들어, p-니트로페놀 또는 페놀을 촉매로 사용한다.4) 5-fluorouracil in (2R, 5R) -5-hydroxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) mentyl ester (V) Coupling to (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate (VII ). In this coupling reaction, a weak organic acid such as p-nitrophenol or phenol is used as a catalyst.

5)(1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VII)를 활성화시킨 후 아민화 하여 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VIII)를 제조한다. 활성화에는 염화설포닐 화합물을, 아민화에는 암모니아, 알킬 아민, 디알킬 아민 등의 아민 화합물을 사용한다.5) (1'R, 2'S, 5'R) -Mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate (VII) Was activated and then aminated to give (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxyl To prepare the plate (VIII). Sulfonyl chloride compounds are used for activation, and amine compounds such as ammonia, alkyl amines and dialkyl amines are used for amination.

6)(1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VIII)를 환원시켜 2R-히드록시메틸-5S-(5'-플루오로시토신-1'-일)-1,3-옥사티올란(IX)을 제조한다.6) (1'R, 2'S, 5'R) -Mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate (VIII) To reduce 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane (IX).

이하 본 발명을 실시예에 의해 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail with reference to Examples.

실시예1: 타르타르산 (1'R,2'S,5'R)-디멘틸에스테르(II)Example 1 Tartaric Acid (1'R, 2'S, 5'R) -Dymentyl Ester (II)

1l-플라스크(Dean-Stark-Trap장착)에 타르타르산(69.6g), l-멘톨(135.2g), p-톨루엔설폰산(35.6g), 300㎖ Tol, 150㎖ THF를 넣고 6시간 환류시켰다. 그동안 물 약 22g이 생성되었다. 반응액을 상온으로 냉각 시킨후 미리 준비한 5%-NaHCO₃수용액 400㎖에 교반하며 적가하였다. 유기층을 분리하고 반응액을 물 200g으로 다시 세척한 후 용매를 감압하에서 완전히 증류시켜 제거하였다.Tartaric acid (69.6 g), l-menthol (135.2 g), p-toluenesulfonic acid (35.6 g), 300 ml Tol, and 150 ml THF were added to a 1 l - flask (with Dean-Stark-Trap ) and refluxed for 6 hours. In the meantime, about 22g of water was produced. The reaction solution was cooled to room temperature and then added dropwise with stirring to 400 ml of a 5% -NaHCO ₃ aqueous solution prepared in advance. The organic layer was separated and the reaction solution was washed again with 200 g of water, and then the solvent was distilled off under reduced pressure.

실시예 2: (1'R,2'S,5'R)-멘틸글리옥실래이트(III)Example 2: (1'R, 2'S, 5'R) -mentylglyoxylate (III)

상기 잔류물(II)에 메탄올 560㎖를 투입하여 10분간 교반하여 용해시켰다.560 ml of methanol was added to the residue (II) , followed by stirring for 10 minutes to dissolve it.

물 30㎖를 투입하고 승온(60-65℃)후, 과요오드산나트륨 92g을 투입하여 4시간동안 환류한 후 냉각(10℃)시켰다. 반응액을 셀라이트(celite) 위로 여과 한후 메탄올로 세척하였다. 여액을 감압 증류하였다. 잔류물에 톨루엔 550㎖와 5% 소듐티오설패이트 80㎖를 넣고 고체가 녹을때까지 교반한 다음, 유기층을 분리하고 소금물 100㎖로 세척하였다. 유기층에 basic charcoal 10g / 무수 황산나트륨 10g을 투입, 1 시간 교반 후에 여과하였다.30 ml of water was added and the temperature was raised (60-65 ° C.), followed by sodium periodate. 92 g was added to reflux for 4 hours, and then cooled (10 ° C.). The reaction solution was filtered over celite and washed with methanol. The filtrate was distilled under reduced pressure. 550 ml of toluene and 80 ml of 5% sodium thiosulfate were added to the residue, followed by stirring until the solid was dissolved. The organic layer was separated and washed with 100 ml of brine. Basic charcoal 10g / 10g anhydrous sodium sulfate was added to the organic layer, and filtered after stirring for 1 hour.

실시예 3: (2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R) 멘틸에스테르(V)Example 3: (2R, 5R) -5-hydroxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) menthyl ester (V)

1L 플라스크(Dean-Stark-Trap장착)에 상기 반응액(III)을 넣고 아세트산 12g을 넣고 환류 (외부온도 : 140℃)하며 물을 제거 한 다음, 반응액을 감압증류하여 잔류물의양이 520㎖가 되도록 하였다. 1,4-디티안-2,5-디올(IV) 60g을 넣고 2 시간 환류시키며 물을 빼냈다.(약 20g). 반응 혼합물의 온도를 약 80℃로 낮춘 다음 여과하고 내부온도를 0 - 5도로 낯춘후 헥산/TEA(1.2ℓ : 12㎖)를 약 2시간에 걸쳐 투입한 후 약 6시간 상기 온도에서 교반하여 생성된 고체를 여과한 후 톨루엔/헥산(1:3) 40㎖ x 2 로 세척하였다. 생성된 흰색 고체를 감압 건조 시켰다(50-55℃도에서 10시간). mp =93.8℃. [α]²⁵ _D= -45 (c = 0.1, MC).¹H NMR(DMSO-d6) 5.56(d,1H), 4.72(t,1H), 3.1(dd,1H), 2.8(dd,1H), 1.4-2.0 (m,7H), 1.0(m,2H), 0.9(t,6H), 0.8(d,3H). MS; 288(2, M+), 271(3), 139(72, 멘톨 x H₂O), 105((50), 83(100).The reaction solution in a 1 L flask (Dean-Stark-Trap equipped)III), 12 g of acetic acid was added, refluxed (external temperature: 140 ° C.), water was removed, and the reaction solution was distilled under reduced pressure to obtain a residue of 520 ml. 1,4-dithiane-2,5-diol (IV) 60 g was added and refluxed for 2 hours to extract water (about 20 g). The temperature of the reaction mixture was lowered to about 80 ° C., filtered and the internal temperature was reduced to 0-5 ° C., followed by addition of hexane / TEA (1.2 L: 12 ml) over about 2 hours, followed by stirring at the temperature for about 6 hours. The solid was filtered and washed with 40 mL × 2 of toluene / hexane (1: 3). The resulting white solid was dried under reduced pressure (10 hours at 50-55 ℃). mp = 93.8 ° C. [α]²⁵ _D= -45 (c = 0.1, MC).^OneH NMR (DMSO-d 6) 5.56 (d, 1 H), 4.72 (t, 1H), 3.1 (dd, 1H), 2.8 (dd, 1H), 1.4-2.0 (m, 7H), 1.0 (m, 2H), 0.9 (t, 6H), 0.8 (d, 3H) . MS; 288 (2, M +), 271 (3), 139 (72, menthol x H₂0), 105 (50), 83 (100).

실시예4:(1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VII)Example 4: (1'R, 2'S, 5'R) -Mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate ( VII)

1ℓ플라스크에 상기 화합물(V)50g과 MC 500㎖에 메탄설폰산 0.13㎖와 DMF 14.3㎖를 넣고 교반하면서 온도를 5-8℃로 냉각하였다 (외부습기차단). 티오닐클로라이드 13.3 ㎖를 상기 온도를 유지하며 약 1시간동안 적가한 다음 반응조의 온도를 10 - 15℃로 올리며 1.5시간 동안 교반한다. 반응액을 1.5시간 동안 상압에서 증류하였다(약 300㎖).50 g of the compound (V) and 500 ml of MC were added to 0.11 ml of methanesulfonic acid and 14.3 ml of DMF in a 1 l flask, and the temperature was cooled to 5-8 ° C. while stirring (external moisture blocking). 13.3 mL of thionyl chloride is added dropwise for about 1 hour while maintaining the above temperature, and then the temperature of the reaction vessel is raised to 10-15 ° C. and stirred for 1.5 hours. The reaction solution was distilled at atmospheric pressure for 1.5 hours (about 300 mL).

500㎖ 가지가 세개 달린 플라스크에 실릴화시켜 증류한 5-플루오로우라실47.6g과 클로로포름 100㎖, p-니트로페놀 5.74g을 넣고 환류시키는 동안 상기 용액을 30분동안 적가하고 5시간 환류시켰다. 물 50㎖와 TEA 12.2㎖의 혼합물에 상온으로 냉각시킨 상기 반응액을 서서히 적가 한후 30분간 교반하고 유기층을 분리하였다. 용매를 완전히 감압, 증류시킨후 이소프로필아세테이트 75㎖를 넣고 10-15℃에서 5시간 교반후 여과하였다. 생성된 흰색 고형물을 55-60℃에서 6시간 감압 건조시켰다. 55g의 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 얻었다(수율 80%). mp = 220℃(dec.). [α]²⁵ _D= -57.9 (c=1.02, DMF).¹H NMR(DMSO-d6); 8.2(d,1H), 6.3(t,1H), 4.7(t,1H), 3.5(dd,1H), 3.2(dd,1H), 1.4-2.0 (m,7H), 1.0(m,2H), 0.9(t,6H), 0.8(d,3H). MS; 401(5, M+1), 270(26), 189(16), 139(30, 멘톨 x H₂O), 83(100).The solution was added dropwise for 30 minutes and refluxed for 5 hours while refluxing 47.6 g of 5-fluorouracil, distilled and silyzed into a 500 ml eggplant flask, and 100 ml of chloroform and 5.74 g of p-nitrophenol. To the mixture of 50 mL of water and 12.2 mL of TEA was slowly added dropwise the reaction solution, which was stirred for 30 minutes and the organic layer was separated. After completely distilling off the solvent under reduced pressure, 75 ml of isopropyl acetate was added thereto, and the mixture was stirred at 10-15 ° C. for 5 hours and filtered. The resulting white solid was dried under reduced pressure at 55-60 ° C. for 6 hours. 55 g of (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate was obtained ( Yield 80%). mp = 220 ° C. (dec.). [a] ²⁵ _D = -57.9 (c = 1.02, DMF). ¹ H NMR (DMSO-d 6); 8.2 (d, 1H), 6.3 (t, 1H), 4.7 (t, 1H), 3.5 (dd, 1H), 3.2 (dd, 1H), 1.4-2.0 (m, 7H), 1.0 (m, 2H) , 0.9 (t, 6H), 0.8 (d, 3H). MS; 401 (5, M + 1), 270 (26), 189 (16), 139 (30, menthol x H ₂ O), 83 (100).

실시예5:(1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트(VIII)Example 5 (1'R, 2'S, 5'R) -Mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate ( VIII)

100㎖ 플라스크에 상기 화합물(VII) 5.0g , 무수 K₂CO₃3.45 g, 토실클로라이드(Tosyl-Cl)4.76 g을 아세토니트릴 50㎖에 넣고 2시간 환류시켰다. 상온으로 냉각시킨후 암모니아를 1-2시간동안 투입하였다. 염화메틸렌 50㎖와 물 50㎖를 넣고 30분 교반후 유기층을 분리하였다. 용매를 감압증류하고 이소프로필 아세테이트 10㎖를 넣고 30분간 교반후 여과하였다. 생성된 고형물을 감압 건조시켜 4.0g의(1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 얻었다(수율 80%). mp = 220℃(dec.). [α]²⁵ _D= -20 (c=0.07, MeOH).¹H NMR(DMSO-d6); 8.1(b,1H), 6.4(t,1H), 5.8(b,1H), 4.8(dt,1H), 3.5(dd,1H), 3.1(dd,1H), 1.4-2.0 (m,7H), 1.0(m,2H), 0.9(t,6H), 0.8(d,3H).In a 100 ml flask, 5.0 g of the compound ( VII ), 3.45 g of anhydrous K ₂ CO ₃ , and 4.76 g of tosyl chloride (Tosyl-Cl) were added to 50 ml of acetonitrile and refluxed for 2 hours. After cooling to room temperature, ammonia was added for 1-2 hours. 50 ml of methylene chloride and 50 ml of water were added thereto, followed by stirring for 30 minutes to separate an organic layer. The solvent was distilled under reduced pressure, 10 ml of isopropyl acetate was added thereto, stirred for 30 minutes, and filtered. The resulting solid was dried under reduced pressure to give 4.0 g of (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R -Carboxylate was obtained (yield 80%). mp = 220 ° C. (dec.). [a] ²⁵ _D = -20 (c = 0.07, MeOH). ¹ H NMR (DMSO-d 6); 8.1 (b, 1H), 6.4 (t, 1H), 5.8 (b, 1H), 4.8 (dt, 1H), 3.5 (dd, 1H), 3.1 (dd, 1H), 1.4-2.0 (m, 7H) , 1.0 (m, 2H), 0.9 (t, 6H), 0.8 (d, 3H).

실시예 6 : 2R-히드록시메틸-5S-(5'-플루오로시토신-1'-일)-1,3-옥사티올란(IX)Example 6 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane (IX)

K₂HPO₄4.58g을 물 50㎖에 용해시켰다. 상기화합물(VIII) 3.5g과 에탄올 35 ㎖를 넣고 10분간 교반하였다. 미리 제조한 NaBH₄0.7 g/ 7㎖ 물/0.07㎖ 25% NaOH 수용액의 혼합물을 약 2시간에 걸쳐 적가하였다. 반응이 완결되면 30분간 정치 한후 유기층을 분리하였다. 진한 HCl 1㎖로 유기층 용액을 pH 4-4.5로 맞추고, 다시 2N-NaOH 4㎖로 pH 6.8-7.2 사이로 맞추었다. 반응물의 유기용매를 완전히 감압증류 한후 물 20 ㎖를 넣고 톨루엔으로 2번 추출하였다. (IX)가 들어 있는 물 층을 감압증류 한후 플래쉬 칼럼 크로마토그래피(flash column chromatography) (MC/MeOH,95:5)로 분리 하였다. 1.95g의 (-)-FTC를 얻었다(수율 95%). mp =195℃, [α]²⁵ _D= -130 (c=1.02, MeOH).¹H NMR (DMSO-d6); 8.2(d,1H), 7.83(br,1H), 7.59(br,1H), 6.1(m,1H), 5.4(t,1H), 5.2(m,1H), 3.7(m,2H), 3.4(dd,1H), 3.1(dd,1H). MS; 248(1, M+1), 130(26, 5-플로오로시토신+1), 118(95), 87(100).4.58 g K ₂ HPO ₄ was dissolved in 50 mL of water. 3.5 g of the compound ( VIII ) and 35 ml of ethanol were added thereto, followed by stirring for 10 minutes. Pre-prepared mixtures of NaBH ₄ 0.7 g / 7㎖ water /0.07㎖ 25% NaOH aqueous solution was added dropwise over a period of about 2 hours. After the reaction was completed, the organic layer was separated after standing for 30 minutes. The organic layer solution was adjusted to pH 4-4.5 with 1 ml of concentrated HCl and again between pH 6.8-7.2 with 4 ml of 2N-NaOH. After distilling the organic solvent completely under reduced pressure, 20 ml of water was added thereto and extracted twice with toluene. The water layer containing ( IX ) was distilled under reduced pressure, and then separated by flash column chromatography (MC / MeOH, 95: 5). 1.95 g of (-)-FTC were obtained (yield 95%). mp = 195 ° C., [a] ²⁵ _D = −130 (c = 1.02, MeOH). ¹ H NMR (DMSO-d 6); 8.2 (d, 1H), 7.83 (br, 1H), 7.59 (br, 1H), 6.1 (m, 1H), 5.4 (t, 1H), 5.2 (m, 1H), 3.7 (m, 2H), 3.4 (dd, 1H), 3.1 (dd, 1H). MS; 248 (1, M + 1), 130 (26, 5-fluorocytosine + 1), 118 (95), 87 (100).

비교예(실시예 5의 화합물(VIII) 합성을 5-플로오로시토신을 출발물질로 하여 국제 특허공개공보 제 WO95/29174 호에 기재된 방법대로 실시하였다). Comparative Example ( Compound ( VIII ) of Example 5 was synthesized according to the method described in WO 95/29174, using 5-fluorocytosine as a starting material).

100㎖ 플라스크에 상기 화합물(V) 6.7g와 염화메틸렌 67㎖, 메탄술폰산 0.02㎖, DMF 2.0㎖를 넣고 5-10℃로 냉각시켰다. 티오닐클로라이드 1.9㎖를 서서히 적가하고 10-15℃에서 1-2시간 교반한 다음 상압에서 MC 35㎖를 증류하였다. 100㎖ 가지가 세개 달린 플라스크에 3.0g 5-플루오로시토신, 5.3㎖ 헥사메틸디실라잔, 0.02㎖ 메탄술폰산, 10㎖ 톨루엔을 넣고 맑은 용액이 될 때까지 환류한다음 50℃로 냉각시킨후 TEA 3.2㎖를 넣고 상기 (V)의 염소화 반응액을 50℃에서 실릴화시킨 5-플루오로시토신 용액에 서서히 적가한다음 5시간 환류시켰다. 반응액을 1.65㎖ TEA/물 25㎖에 현탁시키고 60분 교반한 후 헥산 27㎖를 서서히 적가하여 상온에서 밤새 교반하였다. 여과하고 이소프로필아세테이트로 세척한 다음 건조시켰다. 3.25g의 흰색고체(VIII)를 얻었다(수율 35%)In a 100 ml flask, 6.7 g of the compound ( V ), 67 ml of methylene chloride, 0.02 ml of methanesulfonic acid, and 2.0 ml of DMF were added and cooled to 5-10 ° C. 1.9 ml of thionyl chloride was slowly added dropwise, stirred for 1-2 hours at 10-15 ° C., and 35 ml of MC was distilled off at atmospheric pressure. Into a 100 ml three-necked flask, add 3.0 g 5-fluorocytosine, 5.3 ml hexamethyldisilazane, 0.02 ml methanesulfonic acid, 10 ml toluene, reflux until a clear solution, and cool to 50 ° C. 3.2 ml was added, and the chlorination reaction solution of ( V ) was slowly added dropwise to a 5-fluorocytosine solution silylated at 50 ° C, and then refluxed for 5 hours. The reaction solution was suspended in 25 mL of 1.65 mL TEA / water, stirred for 60 minutes, and then slowly added dropwise with 27 mL of hexane, followed by stirring at room temperature overnight. It was filtered, washed with isopropyl acetate and dried. 3.25 g of white solid (VIII) were obtained (yield 35%).

mp = 220℃(dec.). [α]²⁵ _D= -20 (c=0.07, MeOH).¹H NMR(DMSO-d6); 8.1(b,1H), 6.4(t,1H), 5.8(b,1H), 4.8(dt,1H), 3.5(dd,1H), 3.1(dd,1H), 1.4-2.0 (m,7H), 1.0(m,2H), 0.9(t,6H), 0.8(d,3H).mp = 220 ° C. (dec.). [a] ²⁵ _D = -20 (c = 0.07, MeOH). ¹ H NMR (DMSO-d 6); 8.1 (b, 1H), 6.4 (t, 1H), 5.8 (b, 1H), 4.8 (dt, 1H), 3.5 (dd, 1H), 3.1 (dd, 1H), 1.4-2.0 (m, 7H) , 1.0 (m, 2H), 0.9 (t, 6H), 0.8 (d, 3H).

상기 본 발명의 상세한 설명 및 실시예에서 알 수 있는 바와 같이, 본 발명의 제조방법은 루이스 산을 사용하지 않으면서도 입체 선택적 반응으로 추가적인 이성질체의 분리 작업이 요구되지 않는다. 또한, 커플링 반응시 염기로 고가의 5-플루오로시토신 대신 가격이 싼 5-플루오로우라실을 사용하고 종래의 방법에 비해 수율이 월등히 높아 경제적인 장점이 있다.As can be seen in the description and examples of the present invention, the preparation method of the present invention does not require the separation of additional isomers in a stereoselective reaction without using Lewis acids. In addition, inexpensive 5-fluorouracil is used as a base instead of expensive 5-fluorocytosine in the coupling reaction, and the yield is much higher than that of the conventional method.

Claims (7)

a)(2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R)멘틸에스테르에 5-플루오로우라실을 커플링시켜 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조한 다음,a) 5-fluorouracil is coupled to (2R, 5R) -5-hydroxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) mentyl ester Prepare (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate, b)상기 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란 -2R-카르복실래이트를 활성화시킨 후 아민화하여 (1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 제조하고,b) activating the (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorouracil-1'-yl) -1,3-oxathiolane-2R-carboxylate And then aminated to (1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate Manufacturing the c)상기 (1'R,2'S,5'R)-멘틸 -5S-5- (5'-플루오로시토신-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 환원시키는 것으로 구성되는 것을 특징으로 하는 2R-히드록시메틸-5S-(5'-플루오로시토신 -1'-일)-1,3-옥사티올란의 제조방법c) reducing the (1'R, 2'S, 5'R) -menthyl-5S-5- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane-2R-carboxylate Method for producing 2R-hydroxymethyl-5S- (5'-fluorocytosine-1'-yl) -1,3-oxathiolane, characterized in that consisting of 제 1 항에 있어서, 상기 단계 a)의 출발물질인 (2R,5R)-5-히드록시-[1,3]-옥사티올란-2-카르복실산(1'R,2'S,5'R)멘틸에스테르는, i) 타르타르산(I)에-멘톨과 p-톨루엔설폰산을 반응시켜 (1'R,2'S,5'R)-디멘틸에스테르를 제조한 다음, ii)타르타르산(1'R,2'S,5'R)-디멘틸에스테르를 과요오드화나트륨을 사용하여 (1'R,2'S,5'R)-멘틸글리옥실래이트로 만들고 iii)(1'R,2'S,5'R)-멘틸글리옥실래이트를 1,4-디티안-2,5-디올과 반응시켜 제조하는 것을 특징으로 하는 방법.The (2R, 5R) -5-hydroxy- [1,3] -oxathiolane-2-carboxylic acid (1'R, 2'S, 5'R) according to claim 1, which is a starting material of step a). Menthyl ester is i) to tartaric acid (I) (1'R, 2'S, 5'R) -dimentyl ester was prepared by reacting menthol with p-toluenesulfonic acid, followed by ii) tartaric acid (1'R, 2'S, 5'R) -dimentyl ester. (1'R, 2'S, 5'R) -Mentylglyoxylate was prepared using sodium periodide and iii) (1'R, 2'S, 5'R) -mentylglyoxylate was prepared using 1,4-dithiane. Prepared by reacting with -2,5-diol. 제 2 항에 있어서, 상기 단계 i)의 용매는 공비가 가능한 혼합용매인 것을특징으로 하는 방법.The method of claim 2, wherein the solvent of step i) is azeotropic mixed solvent. 제 3 항에 있어서, 상기 혼합용매는 톨루엔, 벤젠, 및 에틸아세테이트 중 하나와 테트라히드류 퓨란, 아세토니트릴 중 하나의 혼합용매인 것을 특징으로 하는 방법.The method of claim 3, wherein the mixed solvent is a mixed solvent of one of toluene, benzene, and ethyl acetate, and one of tetrahydric furan and acetonitrile. 제 1 항에 있어서, 상기 단계 a)의 커플링 반응은 약 유기산을 촉매로 사용하는 것을 특징으로 하는 방법.The process of claim 1, wherein the coupling reaction of step a) uses a weak organic acid as a catalyst. 제 5 항에 있어서, 상기 약 유기산은 p-니트로페놀 또는 페놀인 것을 특징으로 하는 방법.6. The method of claim 5 wherein the weak organic acid is p-nitrophenol or phenol. 제 1 항에 있어서, 상기 단계 b)의 아민화 반응은 1'R,2'S,5'R)-멘틸-5S-5-(5'-플루오로우라실-1'-일)-1,3-옥사티올란-2R-카르복실래이트를 염화설포닐화합물로 활성화시킨 후 아민과 반응시키는 것으로 구성되는 것을 특징으로 하는 방법.The process of claim 1, wherein the amination reaction of step b) is 1'R, 2'S, 5'R) -mentyl-5S-5- (5'-fluorouracil-1'-yl) -1,3- Oxatiola-2R-carboxylate is activated with a sulfonyl chloride compound and then reacted with an amine.