TW219932B - - Google Patents

Description

219982 V. Description of the invention (1) Background of the invention (1) Field of the invention The present invention relates to an optically active H- (D-Ct-alkyl- / 3-hydrothiothiopropionaldehyde) -L-proline acid The system of law. (2) Description of the related art The N-D-ct-methyl- / 3-hydrothiopropyl propanyl group represented by the following formula (IV) has been found-L-g namamine (common name: captopril Profit (C apt 〇pr ί 丨)):

CH 3

HS-CH2-CH-CON

COOH (IV) Read in Contrary to the meaning of the manuscript η Installed by the Ministry of Economic Affairs, the Bureau of Standards, and only cooperated with the Consumer Council to produce Du Yin. Due to the strong inhibitory effect on the conversion of vasopressin, and based on this inhibitory effect, Therefore, it has a significant blood pressure lowering effect, so it is extremely effective as a blood pressure lowering agent (Biochemistry, 16, 5 4 8 7 (1 9 7?)). As for the preparation method of this N- (D-ct-alkyl- / 3-hydrothiothiopropyl) -L-proline acid, it is known that the optically active D-ct-alkyl- / 3-drug based sulfur Propionic acid halide as a starting material, react with L-amidoamine in an organic solvent or aqueous medium to separate N-(D-ex -alkyl-/?-Certified sulfur! 'T propylene group ) -L-proline, and subsequently, the compound thus separated is subjected to a desulfurization reaction, and then, as desired, N- (D- < α-alkyl- / 3-hydrogen sulfide is obtained Kiblick

L proline. The countermeasures carried out in the organic solvent "reported to be published in Japan No. 4 0 6 7 6/1 9 8 0; while the countermeasures in aqueous media were reported in the Japanese Open Publication No. 1 8 9 8 5/1 9 8 No. 1. However, the acid halide as the starting material is unstable to water. Unless the 5k scale of this book is used in the middle of the 8 standard (CNS) T 4 specifications (210X297 male dragon) 81. 4. 10,000¾ (H) 玎 线 3 219932 cooperation by the employees of the Central Standards Bureau of Japan. Spit_print ¾ V. Description of the invention (2) As invented, the reaction between the acid halide and L-proline (aldimination) should be carried out carefully, otherwise the hydrolysis of the acid halide will occur, resulting in D -By-product of ct-alkyl- / 3-aldehyde thiopropionic acid, which is a hydrolysate of acid halide. Therefore, organic solvents that are inert to acid halides, such as halogenated hydrocarbons and tetrahydrofuran, are generally used as the reaction medium for the amidation reaction. Although the use of these organic solvents can prevent the hydrolysis of acid halides, there is a problem that the solubility of L-proline in these organic solvents is extremely low. Therefore, it is inevitable that the L-proline acid must be suspended in a solid form to perform the amine it reaction and sacrifice the reaction efficiency, or follow the cumbersome reaction process: first convert L-proline acid to this organic solvent Only after the soluble esters are reacted. In the aforementioned amination reaction, a condensing agent for deacidification is required to remove the hydrogen halide generated from the halide. However, when using organic solvents as the reaction medium, it is not allowed to use cheap and economic alkali hydroxides, such as sodium gas oxide. Therefore, it is inevitable to use expensive organic amines, such as Η, Ν-dimethyl Aniline, because organic amines are soluble in organic solvents. This organic amine must be separated and recovered from the reaction product, cetylamine. However, the separation and recovery of organic compounds are more difficult than that of alkali metal hydride. As a result, unreacted organic compounds are easily mixed into the product and become impurities. When carried out in an aqueous medium, the Scotch Bowen (S ch tte η-B auma η η.) Reaction between the acid halide and L-hexylamine St does not appear as if it is carried out in an organic solvent. Problem, but there will be new problems of acid halide hydrolysis. In the method described separately in No. 1 8 9 5 8/1 9 8 No. 1 of Yu Yueben, the acid halide is dissolved in an organic solvent, such as tetrahydrofuran, and the resulting solution is added with L-proline Aqueous solutions to avoid hydrolysis reactions. (Please read the notes beforehand, Sun fills in this page) Packing · Ordering · Thread · The standard of this paper is easy to use in a sleepy home standard (CNS) T4 specification (210x297 public) 81. 4. 10,000¾ (! [) 4 219932 A 6 I? 6 Printed by the Central Bureau of Tests of the Ministry of Examination and Testing 3 The Cooperative Society of Medical Treatment 5. V. Description of the invention (3) However, this method uses organic solvents, therefore, complex post-processing is required to obtain the reaction The organic solvent is separated from the aqueous solution of the product. In addition, various by-products, such as the acid halide hydrolysate, and the reaction product formed by the further reaction of the Scottish Bowman reaction product, usually appear easily in the Scottish Bowman reaction. When the Gordon Baumann reaction product with an acyl group is subjected to a deacylation reaction to obtain the final product with a hydrosulfide group according to the present invention, unless these by-products are removed in advance, the final product is water-soluble Therefore, purification of the final product will become difficult. Therefore, there is no solution unless the following steps are followed: Once the Scotten Bowman product has been purified and separated, the de-arylation reaction is carried out, and the de-acylation product thus obtained is purified. SUMMARY OF THE INVENTION An object of the present invention is to provide a method for preparing L-proline acid derivatives with high yield of M, the method comprising performing a Scotten Bowman reaction of an acid halide and L-proline acid in an aqueous medium At the same time, the hydrolysis of the acid halide is inhibited, thereby performing the deacylation reaction without separating the Scottish Bowman reaction product from the reaction mixture. In view of this, the present inventors have determined the reaction parameters that may affect the yield of the acid halide and L-proline by the Scotton Bowman reaction to the amidation product, as well as the D-ct-alkyl- / 3- Acetyl thiopropionic acid and N-acyl-L-proline and other by-products were thoroughly studied. As a result, it was found that the use of a deacidification condensing agent to control the pH of the reaction mixture to a predetermined level can avoid the hydrolysis of the Xue halide without using an organic solvent, while allowing the amidation reaction to proceed quickly; It is easy to produce by-products under extremely high Η, such as Ν- 醯 基 (please read the precautions before filling in the wooden page) Binding · Binding-Line-This paper standard is not in accordance with the a® family standard (CNS) T4 specifications (210x297 male dragon) 81. 4. 10,000 ^ (11) 5 219932 Λ 6 _Π_6_ V. Description of the invention (4) The formation of -L-proline acid, therefore, the aldination product can be obtained in high yield. It has also been found that the aldination products obtained in the foregoing manner can be directly subjected to deacidification and debasing reactions without prior separation and purification from the reaction mixture. Therefore, N-(D-α-can be obtained in high purity and with high yield Alkyl- / 3-hydrothiothiopropenyl s) -L-proline acid, as a result, the present invention was completed. Therefore, the present invention provides a preparation of the L-proline derivative represented by formula (I):

(Please read the notes on the back of Erguo to write wooden pages first). Installed by 3 Department of Standards Bureau of Ayang, and cooperated with Du Yin ¾ [where R2 stands for low-grade embankment], the method includes the order (I ) Represented by D-cr-; emulsyl-deyl thio (7¾ S3 ® compound: R2 T r1s-ch2-ch-cox (technical) [where R: stands for chrysanthemum, X stands for halogen and R2 It has the above definition: react with L-proline. The method includes:-«-alkyl-/?-Alkyl thiopropionic acid halide (I) added to L-proline-containing radon and In the aqueous medium of the condensing agent used for the removal of ib, the D-α-alkyl- / 3-shodothiopropionic acid halide and the L-amino acid are reversed, while maintaining the reaction temperature below 1 2 Ό And maintaining p Η within the range of 10.5-11.5, thereby obtaining the compound represented by the formula (H): _ _ line _ the paper standard is used in the 8-sleeper standard (CNS) T4 specifications (210x297 male dragon) δ 81. 4. 10,000 ^ (11) 219982 Λ β 15 6 By the Ministry of Economic Affairs of the Ministry of Economic Affairs, Beigong Xiaotuo Cooperative Society, India 4 V. Description of the invention (5) r2 RiS-ch2- (! h-con ^) (II) COOH [where 1 ^ and 1? 2 have the definition as described above]; and then, make The compound (Π) is defermented under strong alkali conditions without separating the compound (3) from the reaction mixture. (D-cx -alkyl_ / 3 -acylthio according to the method of the present invention The formation of by-products of propionic acid and N-acyl-L-proline can be suppressed in the amidation reaction, so N- (Da -alkyl- / 3 -acylthio can be obtained in high purity and high yield Propionyl) -L-proline. Again, according to the method of the present invention, by subjecting the aforementioned product to deacylation, D-ct-alkyl- / 3-hydrothiothiopropionic acid and The formation of by-products such as N-acyl-L-proline, so that H- (D- < α-alkyl- / 3-hydrothiothiopropionyl) -L-pro Aminic acid. The details of the preferred certified examples are shown in formulas (I) and (I). Examples of acetyl groups represented by ^ include C2_7 alkyl acetyl groups, such as acetyl and propyl groups; and aryl acetyl groups , For example, unsubstituted or composed of one or more lower alkyl groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl and tertiary butyl, or halogen atoms (fluorine, chlorine, bromine Or iodine atom), the substituted benzylidene group. In formula (I ), (I) and (S) in the example of lower alkyl represented by r2, including linear or branched _ (: Bu 7 alkyl, such as methyl, ethyl, propyl and isopropyl. Formula (I ), The examples of halogen atoms represented by X include atmosphere, bromine or iodine atoms. (Please read the notes before filling in and then fill out this page) Binding. Thread · This paper standard uses the Chinese National Standard (CNS ) < P4 specification (210x297 g; Ϊ) 7 81. 4. 10,000 ^ (1!) 219932 Λ β Η 6 Ministry of Economic Affairs, Pyongyang Bureau of Industry and Commerce cooperation Du Yin 3i

5. Description of the invention (6) The method for preparing the acid halide (I) of the present invention, for example, through the use of a halogenating agent, such as sulfenamide, phosphorus tribromide or phosphorus tribromide, known by the industry It is obtained by processing the corresponding carboxylic acid (obtained by the method shown in Japanese Patent Publication No. 222798/1989). As for the aqueous medium that can be used in the practice of the present invention, an aqueous solution having a pH of 10.5-11.5 can be used, and a preferred example of a condensation agent for deacidification includes alkaline compounds soluble in an aqueous medium, such as alkali Metal hydroxides, such as sodium hydroxide and potassium hydroxide. The condensing agent used for deacidification can also be used as a pH control agent for aqueous media. In the practice of the present invention, the reaction between the acid halide and L-proline is carried out by adding the acid halide to the aqueous medium containing L-proline, preferably under stirring. During the reaction, the P Hi% of the reaction mixture was maintained in the range of 10.5-11.5. The alkali metal hydroxide is used as a condensation agent for deacidification to remove the hydrogen halide emitted from the acid halide, and it is also used to control the pH of the reaction mixture. It is not appropriate to add the aqueous solution of L-proline to the acid halide, because the residence time of the acid halide in the aqueous solution is too long, and therefore hydrolysis of the acid halide occurs. The pH value of the mixture with the lanthanum becomes lower than 10.5. In other words, when approaching the acidic side, the acid halide becomes more sensitive to hydrolysis. The closer ΡΗ is to ρΗ7, the more D-a-alkyl- / 3-dartylthiopropionic acid by-product, that is, the hydrolysis product, is generated. Therefore, a pH lower than 10.5 is not suitable. On the other hand, as the pH of the reaction mixture rises above 1 1.5 and approaches 14, the more N-scaproyl-L-proline is produced. Therefore, P Η is higher than 1 1.5 & unsuitable. At higher ρ Η, the yield of by-products of N-acyl-L-proline is larger, which may be pseudo-attributed to the produced Η-(D (please read back and note beforehand # 塡 写 此 页) Install. Line, this paper is fully equipped with Chinese B standard (CNS) 4 specifications (210x297 public address) 81. 4. 10,000¾ (II) 8 2ί9982 A fi n 6 Central Ministry of Economic Affairs and β work consumption Cooperation Du Yin ft'14 V. Description of the invention (7) -α-alkyl- / 3-acylthiopropyl (propyl) -L-proline acid reacts with L-proline acid contained in the reaction mixture Reason. It is necessary to perform the amidation reaction at a temperature of 12f or less, preferably 1010 or less, and more preferably 81 or less. A temperature higher than 1 2 13 leads to an increase in the production of by-products of N-acyl and -L-proline, which is not suitable. In this way, it is preferable to keep the reaction mixture at a temperature of 0 T or above while the reaction is proceeding, so as to avoid undesired freezing of the reaction mixture. There is no special limitation on the reaction time, but the reaction time is preferably 0.5-8 hours, more preferably 1-5 hours. Economically, the feed amount of L-proline is advantageously 0.9-1.1 moles / mole acid halide. Although there is no particular limitation on the concentration of the L-proline acid aqueous solution fed, from the viewpoint of controlling the reaction temperature, it is preferably 1-5 0 w 15C, more preferably 2-3 0 wt S; Concentration feed. Alkyl- / 3-acylthiothiopropylbenzyl) -L-proline acid produced by the reaction of acid halide and L-proline acid can be removed after separation and purification in a manner known in the industry Rhenylation reaction. However, in the method of the present invention, the content of by-products in the reaction mixture obtained by the amidation reaction is extremely low. Therefore, the amidation product can be subjected to the deboration reaction in the same reactor, while maintaining the undesired mixture in the alkali Under aerobic conditions, it is not necessary to separate the amidation product after the amidation reaction is substantially complete. As used herein, the expression "after the amidation reaction is substantially completed" means that the amount of at least one of the acid halide and L-proline acid fed to the feed is reduced to 5% or less status. Although there is no special limit to the temperature range of the decyclization reaction, as long as the reaction (please read first, please pay attention to this item, please write this page). Install-Line-Use this standard as a national standard (CNS) A 4 specifications (210x297 g: ¢) 81. 4. 10,000¾ (H) 9 .year Β2.

Printed 2199S2 A6 _; _B6__ by the WBC Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the invention (8) It can be carried out under alkaline conditions. But to shorten the reaction time, it is better to increase the reaction S degree to more than 151. It is better to rise to 20¾ or above. It is better to carry out the reaction at a temperature between 40¾ and 50t: and a pH between 10.5 and 11.5. If the reaction mixture is maintained at a strong pH of 12 or above, the dezymation reaction can be carried out even at lower temperatures. However, economically, these lower temperatures are not beneficial because after the wake-up reaction To extract L-proline derivatives from the reaction mixture will require a large amount of mineral acid. On the other hand, a pH lower than 10.5 will cause problems with more by-products. Therefore, the pH is not suitable outside the aforementioned range. The reaction time required for the decyclization reaction is usually in the range of 10 minutes to 10 hours, preferably 0.5-4 hours. In addition, it is preferably carried out under an inert gas, such as radon, gas and argon atmosphere, and preferably under a gas flow, to carry out the debasing reaction. After the reaction is complete, add mineral acid, such as hydrochloric acid, to adjust the reaction mixture to be acidic. Then, the generated N- (D-cr-alkyl- / 3-hydrothiopropylpropionyl) -L-proline acid is extracted with an organic solvent, such as dichloromethane and ethyl acetate, and reduced. Dry or recrystallize-by which the Sassafras product can be isolated and obtained. The details of the present invention will be explained in more detail in the following examples. However, it should be borne in mind that the present invention is not limited to or only limited by the following examples. Example 1 9.8 s L-proline was added to 90 m of distilled water. The resulting solution was added to 2N hydrogenated aqueous solution to adjust the pH to 11. The resulting aqueous solution was cooled by ice-water bath to 5T: ~ 0C. Maintain the temperature of the reaction mixture at 5C ~ 0t :, add 1 .0 g D-α-methyl-zhao-acetothiothiopropionate dropwise over 1 hour (please read the notes on the back before filling this page ) —Installed-• V-0. This paper scale is applicable to China National Apple (CNS) A 丨 specifications (210 X 297 public loan) jq Jiang Zhengbin 82. 5. 20,000 Jingji Ministry of Central Bureau of Standards and Staff Respiratory Cooperation Th seal? Announcement 丨 Λ6 ......! _Β6__ V. Description of the invention (9) Fermentation atmosphere. After the dropwise addition was completed, the reaction mixture was reacted for another 2 hours. At the beginning of the dropwise addition, the reaction started to proceed and ρΗ began to decrease. Therefore, during the reaction, 2 NH sodium hydroxide aqueous solution was added to the pump by pH control, so that the pH of the reaction mixture was always maintained at 11. After no more D-α-methyl- / 3-acetyl thiopropionaldehyde gas was detected in the anti-mixture, the reaction mixture was heated to 50 ° C in a warm water bath under a nitrogen flow. As the temperature of the reaction mixture rises, its pH begins to decrease again. «The pH of the solution was maintained at 1 1 by adding a 5N hydrogenated sodium aqueous solution via a pH-controlled pump, and the debasing anti-plaque was carried out for another 2 hours. After the deacylation reaction was completed, the molar yield of the reaction product in the reaction mixture was analyzed by high performance liquid chromatography.结果 不 于 表 1。 The results are not in Table 1. Using D-ct-methyl- / 3-benzylthiopropion S atmosphere in equal molar amounts to replace D-α-methyl-dot-acetoxythiopropionase atmosphere can also be carried out with similar success The enzymatic amination and deamination reaction results in the formation of the corresponding L-proline derivative with high purity and high yield. Comparative Examples 1-4 The anti-plaque was carried out in a similar manner as in Example 1, but the -P Η in the S amination reaction was always maintained at 7.6 (in Comparative Example 1, the hydrogenated sodium aqueous solution was replaced with sodium hydrogen peroxide solution) , 10.0 (Comparative Example 2), 12.0 (Comparative Example 3), or at least 13 (Comparative Example 4). The molar yield of the reaction products in each mixture was analyzed by pseudo-hydrazide high performance liquid chromatography. The results are also shown in Table 1. ------------------------ Installed ------, 玎 ------ ^ (Please read the precautions on the back first, then 塡Write this page) This paper scale is applicable to the CNA Standard. !! Regulations (210 X 297 justice), A _ Li 82. 5. 20,000 11 Blowing to buy ^ 19932 V. Description of the invention (10) m_1 Example 1 Comparative example 1 2 3 4 pH of the reaction mixture 11.0 7.6 10.0 12.0 ^ 13 Product (%) 1 95.1 38.7 79.8 85.1 4 0.4 2 3.3 6 1.3 20.2 3.7 22.2 3 1.6 0 0 11.2 37.4 (please read the precautions before writing this page) Table 1 and the following table 2 and Table 3, product 1, product 2 and product 3 respectively represent N- (Da-methyl- / 3-hydrothiopropylpropenyl) -L-proline, Da-methyl- / 9-hydrogen Thiopropionic acid, and N-acyl-L-proline. In Examples 2 and 3, the reaction was carried out in a similar manner as in Example 1, but P Η became 10.5 (Example 2) or 11.5 (Example 3). The molar yield in each reaction mixture was analyzed by high-performance liquid chromatography. The results are shown in Table 2. Table Example 2 Example 3 The pH of the reaction mixture 10.5 11.5; product 1 90.0 92.0: (X) 2 9.8 2.0 3 0.2 1 6 · 0 As can be seen from the results of Examples 1-3 and Comparative Examples 1-4, high purity N- (Dd-Alkyl- / 3-Hydroxythiopropylpropylenyl) -L-proline acid can be obtained in high yields, and its manufacture and assembly-line. #, '| 4 g C rv Pingguo TW Λ _ * Gongyy 12 81. 4. 10,000¾ (H) Λ 6 Π 6 V. Description of the invention (11) The method of carrying out the amidation reaction through pseudo-acidification and simultaneously reacting The pH of the mixture is maintained within the range defined by the present invention, and then, the resulting reaction product is subjected to deacylation under alkaline conditions. Example 4 and Comparative Examples 5 and 6 were reacted in a manner similar to Example 1, but the temperature of amidation was changed. The results are shown in Table 3. (Please read back the precautions before filling in this page) Binding · Order_ Line · Table 3 Example 4 Comparative Example 5 6 Reaction Temperature 10 15 20 (° c) Product 1 9 0.2 83.8 78.7 m 2 7. 8 6. 2 8. 6 3 3.0 ί 10.0 12.7 The Ministry of Economic Affairs, Central Bureau of Industry and Commerce, A Industrial and Consumer Cooperation Du Printing 5 ί Ming Dian T W Λ Λ Reni Gong 81. 4. 10,000? *: (| [) 13 Five Description of the invention (11a) Comparative Example 7 At the end of the warranty period, the amine should be reacted. The production of amine should be reversed except for the example and the application. The reaction should be implemented. JiW The same sulfur 0 乙 I θ · I The purity of the base compound is not 2% 2 8 8 is the rate of production of arginine motilate-f-yl-L alpha-/-yl alpha-fermentation D-propionate propyl sulfide After the acid salt oil N is obtained, if the crystal is removed, the solvent is re-dissolved, the acetic acid vinegar is taken from the extracted alkylate, the formazan oil and gas, and then the acid is removed. Mixed with the base Ν 醯 之 三 銯 漯 单 醯 冯 入入基基 Ηίι A ao% base 7011 for the rate of sulfur production of acetylenic acid mono- Ο I acid amine A-α preserved in the insurance The mixture of Η Ρ 0 should be mixed with the mixture of nitrogen and acid under the temperature of the acid amine, and the temperature of the mixture should be reversed with V 0 5 to the mixture of hot additives. The temperature of the bath water should be 0 WU. Acetate P off 0 and go on and off at the same time. Open 1 degree and then hold the pH to keep its pH. The solubility of the liquid will be lowered to the end to complete 〇Η 化 ^ 0 Ν- take off (please read the notes on the back Then fill in the wood S) Pack · Order _

No. ¾. The 67 fluid is the dissolution rate and should be reversed, which is the fruit of the fruit. Liquid rate rapid production high ears 0 Mo-the total success of the J Leptopka low phase line _ Printed by the Central Standards Bureau of the Ministry of Economic Affairs Beigong Consumer Cooperative Paper standard using the Chinese a standard (CNS) A4 specifications (210x297g *) 13a

Claims (1)

219932 A7 B7 C7 D7 82,12 · 17 Xiuben Ben Six, patent application scope 1. A method for preparing L-amido acid derivative represented by formula (Μ) R 2 HS-CH2-CH-CON COOH (HI) [where L represents a linear or branched C17 alkyl group], assuming that D alkyl- / 3 aldehyde thiopropionic acid halide represented by formula (I) I 丄-/: \ 2 R1S-CH2-CH-COX (I) [In the formula, R i represents C2_7 alkanoyl or unsubstituted or substituted by one or more than two (: 17 alkyl or halogen atom substituted aromatic enzyme, X represents halogen Atom and L have 1 to 4 2 as defined above] react with L-proline, the method includes: adding D-Ot-alkyl- / 3-secretiothiopropionic acid halide (I) to In the aqueous medium of L-pro acid and the water-soluble sickle compound used for deacidification, D-oc-alkyl-/?-Acyl thiopropionic acid halide is reacted with L-proline acid and the reaction The temperature is maintained at 12 = below and the pH is maintained in the range of 10.5-11.5, thereby obtaining the compound represented by formula (E): ------------------- ------ ^ ------. 玎 ------ Line (please note the back of Xianqi first, and then write this page) Printed by the Ministry of Economic Affairs, Zhongji Bureau of Standards, Stomach Consumption, Hefei Society '' 11 ' (II) [In the formula, \ and 1? 2 have the definition as mentioned above]; and without compound (H) being separated from the reaction mixture, the compound should be given to the paper ruler; t applies to the tras house 丨 ticket standard (CNS) A! Specification (210 X 297 X 犮) 1 ^ 10932 Α7 Β7 C7 D7 Six. The scope of patent application (π) is under strong sickle conditions and 2. If the patent application is based on the thiopropionic acid halide, it is held at 10 or below 3. If the patent application is applied, the soluble basic compound is 4. If the patent application is applied, the hydrogenation is sold. 5. If the patent application range is 0.9 to 1.1 moles (I). 6. If the patent application ranges from 401C to 50C and 7. If the patent application is from the following formula (IV))-L-protonic acid: temperature is 15-5 0 ° C. Enterolysis defermentation reaction . The method according to item 1, wherein the reaction between D-a-homoyl- / 3-enzyme (I) and L-proline is carried out under the condition of the reaction temperature. The method around item 1, wherein the water used for deacidification is alkali metal krypton gasification. The method according to item 3, wherein the alkali hydrogenation method is the method according to item 1, wherein the use of L-proline / mole D-cc-alkyl- / 3-cyanothiopropionic acid The method according to item 1 of the tooth circumference, wherein the deacylation reaction P H 1 0.5 to ρ H 1 1.5 is performed. The method around item 1, wherein the L-proline derivatization is represented by H- (D-ct-methyl-zhao-arginylthiopropanyl ch3 HS-CHo-CH-CON COOH (IV) ------------------------- s ------ '玎 ------ ^ (Please read the back first (Notes to write a clothing page again) " ^ Printed by the Ministry of Standards and Staff ’s Consumer Cooperative ^ Only the paper size is applicable to the China S Yuanjia Rubbing Standard (CNS) A 4 specifications (210 X 297 g; $) 2