1245753 欢、發明說明： 【發明所屬之技術領域】 本發明係關於一種合成雙環四氫噻唑乙内醯胨 / γ\ * · cyclic thlaz〇iidine hydant〇in)化合物的方法，特 別疋一種應用在生物素合成領域，以單一反應容器内操作 即可元成兩度結晶性光學活性化合物之簡單方法者。 【先前技術】 生物晶片（biochip)用途，就是將生物的訊號，經過 处里後轉換成可以利用的生物資訊。簡單說，生物晶片 的原理，主要是將單股的DNA、蛋白質（protelns)、抗原 (抓1：1§611)或抗體（抓1：讣〇(^)等作為探針（1^〇1^)，固定在 晶片基材上，晶片材料可能為玻璃片或是尼龍薄膜。將生 物晶片與要檢測的標的物所產生的反應，經由電腦判讀、 比對、分析後，產生有用的資訊。依照生物晶片的特性可 粗分為基因晶片（gene chip)即為DNA微陣列（DNA miCr〇array)、晶片實驗室Gab-on-a-Chip)及蛋白質晶片 (protein chip)二大類。DNA晶片對於基因檢測具有快速 分析、準確率高以及可以同時大量檢測基因的優點，因此 在後基因時代（post —gene age)，生物晶片加速帶動人類 基因功能的自動化分析，生物晶片加速帶動人類基因功能 的自動化分析，也使生物晶片成為生技業中的明星產品。 具光學活性的雙環四氫噻唑乙内醯脲（BicycHc thiazoUdme hydantcnn)化合物就是用在蛋白質與核酸 標示材料上化合物生物素的關鍵中間體（intermediate)。 1245753 根據美國專利 US4009172 ’ US4130713 ’ US4337345 ’ US4550075 ， US4732987 ， US4837402 ， US4877882 ， US4937351 ， US5250699 ， US506834 ， US5095118 ，歐洲專 利EP0243734等所揭示之技術皆可經由雙環四氫噻唑乙内 醯腺（Bicyclic Thiazolidine hydantoin)製造生物素， 而製造雙環四氫嗟σ坐乙内酷脲 （Bicyclic thiazolidine hydanto i η)之方 >去浴p 4艮少。 根據文獻 Chem. Ber_ 1 948 年 81 卷 210頁及 Tetrahedron Lett. 1 988年29卷57頁等習知的記載，都 疋由L-胱氨酸二甲基酯二氫氯化物（[—cystinedime1：hyi ester dihydrochloride)為起始物依下式所示合成雙環四 獻口塞口坐乙内酉藍脲 （Μ ] . . vuicyciic tniazolidine hydantoin) 5 見下列反應式所示： hci.h2n 〇 H^rCOOMe ll CCI3COCCI3, PhCH3 jLMHDn 3) Ph3P« 90〇C. 叫 DME/Me0H/H20 )2 ~~~~ ^ H“「C〇〇Me ----——^ - Bn 、St21245753 Description of the invention: [Technical field to which the invention belongs] The present invention relates to a method for synthesizing a bicyclic tetrahydrothiazole hydantoin / γ \ * · cyclic thlaz〇iidine hydant〇in) compound, and particularly to an application in biology In the field of element synthesis, a simple method that can be converted into a two-degree crystalline optically active compound by operating in a single reaction vessel. [Previous technology] The purpose of biochip is to convert the biological signal into biologic information that can be used after passing through it. To put it simply, the principle of biochips is mainly to use single-stranded DNA, proteins (protelns), antigens (grabbing 1: 1§611) or antibodies (grabbing 1: 讣 〇 (^)) as probes (1 ^ 〇1 ^), Fixed on the wafer substrate, the wafer material may be a glass sheet or a nylon film. The reaction generated by the biochip and the target to be detected is generated by computer through interpretation, comparison, and analysis, and generates useful information. According to the characteristics of biochips, gene chips can be roughly divided into two categories: DNA microarrays (DNA microarrays), wafer labs (Gab-on-a-Chip), and protein chips. DNA chips have the advantages of fast analysis, high accuracy, and large number of genes can be detected simultaneously. Therefore, in the post-gene age, biochips accelerate the automated analysis of human gene functions, and biochips accelerate human genes. The automated analysis of functions has also made biochips a star product in the biotechnology industry. BicycHc thiazoUdme hydantcnn compound with optical activity is a key intermediate of the compound biotin used in protein and nucleic acid labeling materials. 1245753 According to the US patents US4009172 'US4130713' US4337345 'US4550075, US4732987, US4837402, US4877882, US4937351, US5250699, US506834, US5095118, European patent EP0243734 and other disclosed technologies can be passed through the bicyclic tetrahydrothiazole hydantoin The method of producing biotin, and the method of producing bicyclic tetrathiazolidine (bicyclic thiazolidine hydanto i η) > Going to bath p 4 is less. According to the conventional records in the literature, such as Chem. Ber_ 1 vol. 81, page 210, and Tetrahedron Lett. 1 988, Vol. 29, page 57, etc., they are all composed of L-cystine dimethyl ester dihydrochloride ([—cystinedime1: (hyi ester dihydrochloride) is the starting material to synthesize a bicyclic four-orifice stopper (U). Vuicyciic tniazolidine hydantoin) 5 See the following reaction formula: hci.h2n 〇H ^ rCOOMe ll CCI3COCCI3, PhCH3 jLMHDn 3) Ph3P «90〇C. Called DME / Me0H / H20) 2 ~~~~ ^ H" 「C〇〇Me -------- ^-Bn, St2
4) 6N HCI, 1〇〇°C4) 6N HCI, 100 ° C
〇 人 HN N〇 People HN N
〇 SH 5) PhCHO, P〇CI3，PhCH3 〇〇 SH 5) PhCHO, P〇CI3, PhCH3 〇
-Bn 〇 上述文獻所述之方法 1 ·反應步驟需5 2 ·反應試劑昂貴 3·廢棄物難 有下列缺點： 步驟，費時太長。 ’且見毒化物特性。 <理’如二苯基膦氧化物 1245753 得到具光學活性雙環四氫“乙内轉（Blcycllc thiazolidme hydantQln)化合物，以期消除或減輕習用 製作方法之缺點。 本么明之主要目的係在於提供一種合成具光學活性雙 衣四氫垂唑乙内醯脲（Biqclic thiazol idme hydar^t01n)化合物的方法，其只需要在同一反應容器内操 作，热需經單離純化中間體，即可環化合成產物，以達到 簡化製作過程之目的者。 本毛明的另一目的係在於提高雙環四氫噻唑乙内醯脲 的生產效率，可縮短反應時間及純化步驟以及避免產物流 失。 本發明之再一目的是提高產品的結晶性。 為了達到前述的發明目的，本發明所運用的技術手段 係在於：其方法係以L-( + )-半胱氨酸（L(+) —cysteine)， S全類與苯曱基異氰酸酯（benzy 1 is〇cyanate)及分子篩添加 劑為反應物依序進行雙環化合成反應，其反應式如下所示-Bn 〇 The method described in the above literature 1 · The reaction step requires 5 2 · The reaction reagent is expensive 3 · The waste is difficult There are the following disadvantages: The steps take too long. 'See also the properties of poisons. < Li 'such as diphenylphosphine oxide 1247553 to obtain an optically active bicyclic tetrahydro "B internal transfer (Blcycllc thiazolidme hydantQln) compound, in order to eliminate or alleviate the shortcomings of conventional manufacturing methods. The main purpose of this Meming is to provide a synthetic A method for optically active Biqclic thiazol idme hydar ^ t01n compound, which only needs to be operated in the same reaction vessel, and the intermediate needs to be purified by isolation and then cyclized to synthesize the product. In order to achieve the purpose of simplifying the production process, another objective of the present invention is to improve the production efficiency of bicyclotetrahydrothiazole hydantoin, which can shorten the reaction time and purification steps and avoid product loss. Another object of the present invention To improve the crystallinity of the product. In order to achieve the aforementioned object of the invention, the technical means used in the present invention lies in that the method is based on L-(+)-cysteine (L (+)-cysteine), all types of S Sequential bicyclization reaction with benzyl isocyanate (benzy 1 is〇cyanate) and molecular sieve additives as reactants, the reaction formula is shown below
R1ch〇 + r2nc〇 (Additive)R1ch〇 + r2nc〇 (Additive)
C5的烷基、笨 其中R1 ’ R2代表氫原子或苯環或苯曱基 環或苯化C广C5的烷基等取代基。 操作上述反應包含有下列步驟： 1245753 半胱氰 l (L—( + )-Cysteine)、酸 _、右德 驗與C广CR有；^十， 、有换 機酵浴劑反應生成白色固體中間體； 去除C广cs有機醇溶劑； 加入固體分子筛添加劑、苯甲基異氰醋 (benzylls〇cyanate)與酮類溶劑攪拌； 去除_類溶劑； 加入醚類溶劑及無機酸攪拌，形成一下層水層I一 層酯層； 去除固體分子篩添加劑及水層； 去除醚類溶劑； 加入C4的醇類溶劑逼晶； 去除的醇類溶劑；與 乾餘’即可得產物白色固體之雙環四氫噻唑乙内醯腺 〇 由方、所有操作可在同一反應容器内進行，可以簡化操 作之程序，（並可加速雙環化反應，縮短反應時間），同時 ’本發明中利用分子篩為添加劑於反應中進行除水的功能 ’使產π口結晶性提高，為一種新穎的提高產品結晶性之方 法。 【實施方式】 本％明之合成光學活性雙環四氫嗟σ坐乙内酿胧 (Bicyclic thiazolidine hydantoin)化合物的方法，其 彡要合成方法請參考如下所列之反應方程式及所得產物：C5 alkyl group, wherein R1'R2 represents a hydrogen atom or a benzene ring or a phenylfluorenyl ring or a substituent such as a C5 alkyl group. The operation of the above reaction includes the following steps: 1245753 Cysteine (L — (+)-Cysteine), acid, and dextran are compatible with the C-CR; ^ ten, there is a change of the enzyme bath agent to form a white solid intermediate Removal of organic solvents from C and Cs; adding solid molecular sieve additives, benzyl isocyanate (benzylls cyanate) and ketone solvents; removing _-type solvents; adding ether solvents and inorganic acids and stirring to form a lower layer of water Layer I and ester layer; removal of solid molecular sieve additives and water layer; removal of ether solvents; addition of C4 alcohol solvents to force crystals; removal of alcohol solvents; Inner gland: all the operations can be performed in the same reaction container, which can simplify the operation process (and can accelerate the bicyclization reaction and shorten the reaction time). At the same time, in the present invention, molecular sieve is used as an additive to remove the reaction. The function of water improves the crystallinity of the product, which is a novel method to improve the crystallinity of products. [Embodiment] A method for synthesizing an optically active bicyclic tetrahydropyrazine sigma bicyclic compound (Bicyclic thiazolidine hydantoin). The synthesis method please refer to the reaction equations and products obtained below: