CN102796165B

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Info

Publication number: CN102796165B
Application number: CN201110135588.8A
Authority: CN
Inventors: 赵明; 彭师奇; 于言言
Original assignee: Capital Medical University
Current assignee: Capital Medical University
Priority date: 2011-05-24
Filing date: 2011-05-24
Publication date: 2014-02-26
Anticipated expiration: 2031-05-24
Also published as: CN102796165A

Abstract

The invention discloses Arg-Tyr modified aliphatic amine/ alcohol derivatives and a preparation method thereof and application thereof as anodynes and anti-inflammatory agents. The structure of the Arg-Tyr modified aliphatic amine/ alcohol derivatives is shown as a general formula I, namely Arg-TyrXCH2(CH2)nCH3, wherein in the formula, X is NH or O; and n is 6, 8, 10, 12, 14 or 16. The pain relieving activity of the compounds in the general formula is evaluated by adopting a mouse heat thrilling process model, the anti-inflammatory activity of the compounds in the general formula I is evaluated by adopting a mouse ear swelling model, and experimental results show that the 12 compounds represented by the general formula I have excellent pain relieving effect and high anti-inflammatory activity under the same dose and can be applied to production of analgesics and anti-inflammatory medicaments.

Description

Translated fromChinese

京都酚逆序列修饰的脂肪胺/醇衍生物及其制备方法和应用Fatty amine/alcohol derivatives modified by reverse sequence of Kyoto phenol, preparation method and application thereof

技术领域technical field

本发明涉及一种人工合成的多肽化合物，特别是涉及一种对二肽的羧基端用脂肪胺、脂肪醇进行保护修饰而得到的化合物及其制备方法以及在制备镇痛抗炎药物中的应用。The present invention relates to an artificially synthesized polypeptide compound, in particular to a compound obtained by protecting and modifying the carboxyl end of dipeptide with fatty amine and fatty alcohol, its preparation method and its application in the preparation of analgesic and anti-inflammatory drugs .

背景技术Background technique

疼痛作为人类主要生命指征之一，具有重要的生物学意义。世界疼痛大会将疼痛确认为继呼吸、脉搏、体温和血压之后的“人类第5大生命指征”，众多患者正在忍受着疼痛的折磨。目前，临床上使用的镇痛药物都存在各自的毒副作用，例如阿片类镇痛药的成瘾性和水杨类镇痛药对胃肠道的刺激性，寻找镇痛药物是新药研究的热点之一。在镇痛剂设计中发明人认识到京都酚逆序列(Arg-Tyr)是一种有潜力的镇痛二肽，以及口服活性差。按照这种认识，发明人对二肽的羧基端用脂肪胺、脂肪醇进行保护修饰，形成了本发明。As one of the main vital signs of human beings, pain has important biological significance. The World Pain Conference recognized pain as the "fifth vital sign of human beings" after breathing, pulse, body temperature and blood pressure, and many patients are suffering from pain. At present, the analgesics used clinically have their own side effects, such as the addiction of opioid analgesics and the irritation of salicylic analgesics to the gastrointestinal tract. Finding analgesics is a hot spot in the research of new drugs one. In the design of analgesics, the inventors realized that Kyoto phenol reverse sequence (Arg-Tyr) is a potential analgesic dipeptide, and its oral activity is poor. Based on this understanding, the inventors protected and modified the carboxyl end of the dipeptide with fatty amines and fatty alcohols to form the present invention.

发明内容Contents of the invention

本发明要解决的第一个技术问题是，提供通式I中的化合物(5a-l)：Arg-Tyr-XCH₂(CH₂)_nCH₃式中，X为NH或O；式中，n为6、8、10、12、14或16。The first technical problem to be solved by the present invention is to provide the compound (5a-l) in the general formula I: Arg-Tyr-XCH₂ (CH₂ )_n CH₃ In the formula, X is NH or O; in the formula, n is 6, 8, 10, 12, 14 or 16.

对所合成的通式I中的化合物(5a-l)进行编号：5a中X＝NH，n＝6；5b中X＝NH，n＝8；5c中X＝NH，n＝10；5d中X＝NH，n＝12；5e中X＝NH，n＝14；5f中X＝NH，n＝16；5g中X＝O，n＝6；5h中X＝O，n＝8；5i中X＝O，n＝10；5j中X＝O，n＝12；5k中X＝O，n＝14；5l中X＝O，n＝16。The compound (5a-l) in the synthesized general formula I is numbered: X=NH in 5a, n=6; X=NH in 5b, n=8; X=NH in 5c, n=10; X=NH, n=12; X=NH in 5e, n=14; X=NH in 5f, n=16; X=O in 5g, n=6; X=O in 5h, n=8; X=O, n=10; X=O, n=12 in 5j; X=O, n=14 in 5k; X=O, n=16 in 5l.

以上编码号只是为了方便描述，并无任何对发明的限制意义。The above code numbers are only for convenience of description, and do not have any limiting meaning to the invention.

本发明所要解决的第二个技术问题是提供通式化合物5a-l的制备方法，该方法包括：The second technical problem to be solved by the present invention is to provide a preparation method of the compound ofgeneral formula 5a-1, the method comprising:

(1)浓硫酸、浓硝酸条件下，Arg硝基化得N^G-Arg(NO₂)；(1) Under the conditions of concentrated sulfuric acid and concentrated nitric acid, Arg is nitrated to^NG -Arg(NO₂ );

(2)弱碱性水二氧六环条件下，N^G-Arg(NO₂)与(Boc)₂O反应制得N^α-Boc-N^G-Arg(NO₂)，L-Tyr与(Boc)₂O反应制得Boc-Tyr；(2) Under the condition of dioxane in weak alkaline water, N^G -Arg(NO₂ ) reacts with (Boc)₂ O to prepare N^α -Boc-N^G -Arg(NO₂ ), and L-Tyr reacts with ( Boc)₂ O reaction makes Boc-Tyr;

(3)在DCC和HOBt存在下，Boc-Tyr在无水THF中与脂肪胺(CH₃(CH₂)_nCH₂NH₂，n＝6，8，10，12，14，16)或脂肪醇(CH₃(CH₂)_nCH₂OH，n＝6，8，10，12，14，16)缩合成Boc-Tyr-XCH₂(CH₂)_nCH₃；(3) In the presence of DCC and HOBt, Boc-Tyr reacted with aliphatic amines (CH₃ (CH₂ )_n CH₂ NH₂ , n=6, 8, 10, 12, 14, 16) or aliphatic amines in anhydrous THF Condensation of alcohols (CH₃ (CH₂ )_n CH₂ OH, n=6, 8, 10, 12, 14, 16) into Boc-Tyr-XCH₂ (CH₂ )_n CH₃ ;

(4)在含氯化氢的乙酸乙酯中，Boc-Tyr-XCH₂(CH₂)_nCH₃脱Boc保护，得Tyr-XCH₂-(CH₂)_nCH₃；(4) In ethyl acetate containing hydrogen chloride, Boc-Tyr-XCH₂ (CH₂ )_n CH₃ is de-Boc-protected to obtain Tyr-XCH₂ -(CH₂ )_n CH₃ ;

(5)在DCC和HOBt存在下，N^α-Boc-N^G-Arg(NO₂)在无水THF中与Boc-Tyr-X-CH₂(CH₂)_nCH₃缩合，制备得到N^α-Boc-N^G-Arg(NO₂)-Tyr-XCH₂(CH₂)_nCH₃；(5) In the presence of DCC and HOBt, N^α -Boc-^NG -Arg(NO₂ ) was condensed with Boc-Tyr-X-CH₂ (CH₂ )_n CH₃ in anhydrous THF to prepare N^α -Boc-N^G -Arg(NO₂ )-Tyr-XCH₂ (CH₂ )_n CH₃ ;

(6)H₂/Pd条件下N^α-Boc-N^G-Arg(NO₂)-Tyr-XCH₂(CH₂)_nCH₃脱-NO₂保护，得Boc-Arg-Tyr-XCH₂(CH₂)_nCH₃；(6) Under the condition of H₂ /Pd, N^α -Boc-^NG -Arg(NO₂ )-Tyr-XCH₂ (CH₂ )_n CH₃ is de-protected by -NO₂ to obtain Boc-Arg-Tyr-XCH₂ ( CH₂ )_n CH₃ ;

(7)在含氯化氢的乙酸乙酯中，Boc-Arg-Tyr-XCH₂(CH₂)_nCH₃脱Boc保护，得Arg-Tyr-XCH₂(CH₂)_nCH₃。(7) In ethyl acetate containing hydrogen chloride, Boc-Arg-Tyr-XCH₂ (CH₂ )_n CH₃ is de-Boc-protected to obtain Arg-Tyr-XCH₂ (CH₂ )_n CH₃ .

该制备方法可以用图1的路线概括。The preparation method can be summarized by the route in Figure 1.

本发明所要解决的第三个技术问题是提供了所述的化合物在制备镇痛和抗炎药物中的应用。The third technical problem to be solved by the present invention is to provide the application of the compound in the preparation of analgesic and anti-inflammatory drugs.

本发明所要解决的第四个技术问题是在小鼠热甩尾法模型上评价本发明的化合物的镇痛活性。The fourth technical problem to be solved by the present invention is to evaluate the analgesic activity of the compound of the present invention on the mouse heat tail flick model.

本发明的第五个目的是在小鼠耳肿胀模型上评价本发明的化合物的抗炎活性。A fifth object of the present invention is to evaluate the anti-inflammatory activity of the compounds of the present invention on the mouse ear swelling model.

附图说明Description of drawings

图1为本发明通式5a-l化合物的合成路线示意图。Fig. 1 is a schematic diagram of the synthetic route of the compound ofgeneral formula 5a-1 of the present invention.

i)DCC，HOBt，CH₃(CH₂)_nCH₂NH₂(n＝6，8，10，12，14，16)或CH₃(CH₂)_nCH₂OH，(n＝6，8，10，12，14，16)；ii)EtoAc/HCl，冰浴；iii)DCC，HOBt，Boc-Tyr；iv)H₂，Pd/C；v)EtOAc/HCl，冰浴；5a中X＝NH，n＝6；5b中X＝NH，n＝8；5c中X＝NH，n＝10；5d中X＝NH，n＝12；5e中X＝NH，n＝14；5f中X＝NH，n＝16；5g中X＝O，n＝6；5h中X＝O，n＝8；5i中X＝O，n＝10；5j中X＝O，n＝12；5k中X＝O，n＝14；5l中X＝O，n＝16。i) DCC, HOBt, CH₃ (CH₂ )_n CH₂ NH₂ (n=6, 8, 10, 12, 14, 16) or CH₃ (CH₂ )_n CH₂ OH, (n=6, 8 , 10, 12, 14, 16); ii) EtoAc/HCl, ice bath; iii) DCC, HOBt, Boc-Tyr; iv) H₂ , Pd/C; v) EtOAc/HCl, ice bath; X in 5a =NH, n=6; X=NH in 5b, n=8; X=NH in 5c, n=10; X=NH in 5d, n=12; X=NH in 5e, n=14; X in 5f =NH, n=16; X=O in 5g, n=6; X=O in 5h, n=8; X=O in 5i, n=10; X=O in 5j, n=12; X in 5k =O, n=14; X=O, n=16 in 5l.

具体实施方式Detailed ways

为了进一步阐述本发明，下面给出一系列实施例。这些实施例完全是例证性的，它们仅用来对本发明进行具体描述，不应当理解为对本发明的限制。In order to further illustrate the present invention, a series of examples are given below. These examples are entirely illustrative, and they are only used to specifically describe the present invention, and should not be construed as limiting the present invention.

实施例1制备N^G-Arg(NO₂)Example 1 Preparation of^NG -Arg(NO₂ )

冰盐浴搅拌条件下，向500ml茄瓶中加入80ml浓硝酸，然后缓慢加入80ml浓硫酸。10min后，分批加入47.33g(272mmol)L-Arg，反应混合物冰盐浴下搅拌反应2h。向反应混合物中缓慢加入冰水96ml，然后缓慢加入浓氨水，调pH＝8，再用冰醋酸调pH＝6，有大量无色固体析出，放入4℃冰箱中继续析晶。减压过滤，弃去滤液，滤饼用80℃热水溶解，放入4℃冰箱中重结晶。室温下减压过滤，收集不溶物52.5g，为无色固体，产率88％.ESI-MS(m/e)：220[M+H]⁺.Under the condition of ice-salt bath stirring, add 80ml concentrated nitric acid to 500ml eggplant bottle, then slowly add 80ml concentrated sulfuric acid. After 10 min, 47.33 g (272 mmol) of L-Arg was added in batches, and the reaction mixture was stirred and reacted for 2 h in an ice-salt bath. Slowly add 96ml of ice water to the reaction mixture, then slowly add concentrated ammonia water, adjust the pH to 8, then adjust the pH to 6 with glacial acetic acid, a large amount of colorless solid precipitates, put it in a refrigerator at 4°C to continue crystallization. Filter under reduced pressure, discard the filtrate, dissolve the filter cake with hot water at 80°C, and put it in a refrigerator at 4°C for recrystallization. Filter under reduced pressure at room temperature to collect 52.5 g of insoluble matter as a colorless solid with a yield of 88%. ESI-MS (m/e): 220[M+H]⁺ .

实施例2制备N^α-Boc-N^G-Arg(NO₂)Example 2 Preparation of N^α -Boc-^NG -Arg(NO₂ )

称取1.752g N^G-Arg(NO₂)于反应瓶中，加入3ml水溶解，冰浴条件下，缓缓滴加1N NaOH 8ml，将溶有1.92g(Boc)₂O的二氧六环缓缓倒入其中，调反应液pH为9，反应48h，期间每隔2h用单通抽反应体系中CO₂一次。TLC检测反应完全后，调反应液pH为7，旋除二氧六环，再调反应液pH为2，用乙酸乙酯萃取反应液三次，得到的乙酸乙酯层再用饱和NaCl萃洗三次，得到乙酸乙酯层用无水NaSO₄干燥过夜，过滤除去NaSO₄，旋干乙酸乙酯得到标题化合物2.4g，为无色固体，产率94％.ESI-MS(m/e)：320[M+H]⁺.Weigh 1.752g^NG -Arg(NO₂ ) into a reaction flask, add 3ml of water to dissolve it, and slowly add 8ml of 1N NaOH dropwise under ice bath conditions, and dissolve 1.92g of (Boc)₂ O in dioxane Slowly pour it into it, adjust the pH of the reaction solution to 9, and react for 48 hours. During this period, the CO₂ in the reaction system is pumped once every 2 hours with a single pass. After the reaction is complete as detected by TLC, adjust the pH of the reaction solution to 7, spin off dioxane, then adjust the pH of the reaction solution to 2, extract the reaction solution three times with ethyl acetate, and then wash the ethyl acetate layer three times with saturated NaCl , the ethyl acetate layer was dried overnight with anhydrous NaSO₄ , NaSO₄ was removed by filtration, and the ethyl acetate was spin-dried to obtain 2.4 g of the title compound as a colorless solid with a yield of 94%. ESI-MS (m/e): 320 [M+H]⁺ .

实施例3制备Tyr-OMeEmbodiment 3 prepares Tyr-OMe

将120ml无水甲醇放入反应瓶中，冰浴搅拌下滴加10.4ml氯化亚砜，30min内滴加完毕，分批加入7.24g(40mmol)L-Tyr，反应混合物室温搅拌24小时，薄层层析监测至原料消失。水泵减压抽干反应液，加入甲醇溶解，抽干，重复3次；加入乙醚，抽干，重复3次。用甲醇-乙醚重结晶，过滤得标题化合物8.78g，为无色固体，产率87％.ESI-MS(m/e)：196[M+H]⁺.Put 120ml of anhydrous methanol into the reaction flask, add 10.4ml of thionyl chloride dropwise under stirring in an ice bath, and dropwise add 7.24g (40mmol) of L-Tyr in batches within 30min, and stir the reaction mixture at room temperature for 24 hours, thin Layer chromatography was monitored until the disappearance of the starting material. The water pump decompressed and drained the reaction liquid, added methanol to dissolve, drained, and repeated 3 times; added ether, drained, and repeated 3 times. Recrystallized with methanol-ether, and filtered to obtain 8.78 g of the title compound as a colorless solid, with a yield of 87%. ESI-MS (m/e): 196[M+H]⁺ .

实施例4制备Boc-TyrEmbodiment 4 prepares Boc-Tyr

称取0.905g Boc-Tyr于反应瓶中，加入2ml水溶解，冰浴条件下，缓缓滴加1N NaOH5ml，将溶有1.199g(Boc)₂O的二氧六环缓缓倒入其中，调反应液pH为9，反应48h，期间每隔2h用单通抽反应体系中CO₂一次。TLC检测反应完全后，调反应液pH为7，旋除二氧六环，再调反应液pH为2，用乙酸乙酯萃取反应液三次，得到的乙酸乙酯层再用饱和NaCl萃洗三次，得到乙酸乙酯层用无水NaSO₄干燥过夜，过滤除去NaSO₄，旋干乙酸乙酯得到标题化合物0.91g，为无色固体，产率65％.ESI-MS(m/e)：282[M+H]⁺.Weigh 0.905g of Boc-Tyr into the reaction bottle, add 2ml of water to dissolve, slowly add 1N NaOH 5ml dropwise under ice bath condition, and slowly pour 1.199g of (Boc)₂ O dioxane into it, Adjust the pH of the reaction solution to 9, and react for 48 hours, during which CO2 in the reaction system was pumped_once every 2 hours with a single pass. After the reaction is complete as detected by TLC, adjust the pH of the reaction solution to 7, spin off dioxane, then adjust the pH of the reaction solution to 2, extract the reaction solution three times with ethyl acetate, and then wash the ethyl acetate layer three times with saturated NaCl , the ethyl acetate layer was dried overnight with anhydrous NaSO₄ , NaSO₄ was removed by filtration, and the ethyl acetate was spin-dried to obtain 0.91 g of the title compound as a colorless solid with a yield of 65%. ESI-MS (m/e): 282 [M+H]⁺ .

实施例5制备Boc-Tyr-NHCH₂(CH₂)₆CH₃ (1a)Example 5 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₆ CH₃ (1a)

将2.256g(8.03mmol)Boc-Tyr溶20ml无水四氢呋喃(THF)，冰浴下往里加1.35g(10mmol)N-羟基苯并三唑(HOBt)和2.472g(12mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟，得对应的活泼酯溶液，待用。Dissolve 2.256g (8.03mmol) Boc-Tyr in 20ml anhydrous tetrahydrofuran (THF), add 1.35g (10mmol) N-hydroxybenzotriazole (HOBt) and 2.472g (12mmol) dicyclohexylcarbonyldi imine (DCC) in anhydrous THF. The reaction mixture was stirred in an ice bath for 30 minutes to obtain the corresponding active ester solution for use.

将1.29g(10mmol)CH₃(CH₂)₇NH₂溶于20ml无水四氢呋喃(THF)，然后与上面待用的活泼酯溶液混溶，用N-甲基吗啉(NMM)调pH＝8～9。得到的反应混合物室温反应24小时。TLC(展开剂CHCl₃∶CH₃OH＝30∶1)显示CH₃(CH₂)₇NH₂消失。反应混合物减压浓缩至干，残留物用乙酸乙酯溶解，滤除不溶物。滤液依次用5％碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次；5％硫酸氢钾水溶液洗3次，饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩，重结晶纯化，得到标题化合物2.0g，为无色固体，产率64％.ESI-MS(m/e)：415.7[M+Na]⁺.Dissolve 1.29g (10mmol) CH₃ (CH₂ )₇ NH₂ in 20 ml of anhydrous tetrahydrofuran (THF), and then mix with the active ester solution to be used above, adjust the pH with N-methylmorpholine (NMM) = 8~9. The resulting reaction mixture was reacted at room temperature for 24 hours. TLC (developer CHCl₃ :CH₃ OH=30:1) showed that CH₃ (CH₂ )₇ NH₂ disappeared. The reaction mixture was concentrated to dryness under reduced pressure, the residue was dissolved in ethyl acetate, and the insoluble matter was filtered off. The filtrate was washed 3 times with 5% sodium bicarbonate aqueous solution successively, and saturated sodium chloride aqueous solution was washed 3 times; 5% potassium hydrogensulfate aqueous solution was washed 3 times, saturated sodium chloride aqueous solution was washed 3 times and saturated sodium bicarbonate aqueous solution was washed 3 times, Wash with saturated sodium chloride aqueous solution 3 times. The separated ethyl acetate layer was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 32°C and purified by recrystallization to obtain 2.0 g of the title compound as a colorless solid with a yield of 64%. ESI-MS (m/e ): 415.7[M+Na]⁺ .

实施例6制备Boc-Tyr-NHCH₂(CH₂)₈CH₃ (1b)Example 6 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₈ CH₃ (1b)

按照实施例5的方法，3.472g(12mmol)Boc-Tyr与1.57g(10mmol)CH₃(C-H₂)₉NH₂反应，得到标题化合物3.1g，为无色固体，产率74％.ESI-MS(m/e)：443.5[M+Na]⁺.According to the method of Example 5, 3.472g (12mmol) Boc-Tyr was reacted with 1.57g (10mmol) CH₃ (CH₂ )₉ NH₂ to obtain 3.1 g of the title compound as a colorless solid with a yield of 74%. ESI- MS(m/e): 443.5[M+Na]⁺ .

实施例7制备Boc-Tyr-NHCH₂(CH₂)₁₀CH₃ (1c)Example 7 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₁₀ CH₃ (1c)

按照实施例5的方法，3.372g(12mmol)Boc-Tyr与1.85g(10mmol)CH₃(C-H₂)₁₁NH₂反应，得到标题化合物3.45g，为无色固体，产率77％.ESI-MS(m/e)：471.5[M+Na]⁺.According to the method of Example 5, 3.372g (12mmol) Boc-Tyr was reacted with 1.85g (10mmol) CH₃ (CH₂ )₁₁ NH₂ to obtain 3.45 g of the title compound as a colorless solid with a yield of 77%. ESI- MS(m/e): 471.5[M+Na]⁺ .

实施例8制备Boc-Tyr-NHCH₂(CH₂)₁₂CH₃ (1d)Example 8 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₁₂ CH₃ (1d)

按照实施例5的方法，5.48g Boc-Tyr(18mmol)与3.215g(15mmol)CH₃(C-H₂)₁₃NH₂反应，得到标题化合物5.81g，为无色固体，产率81％.ESI-MS(m/e)：499.5[M+N4a]⁺.According to the method of Example 5, 5.48g Boc-Tyr (18mmol) was reacted with 3.215g (15mmol) CH₃ (CH₂ )₁₃ NH₂ to obtain 5.81g of the title compound as a colorless solid with a yield of 81%.ESI- MS(m/e): 499.5[M+N4a]⁺ .

实施例9制备Boc-Tyr-NHCH₂(CH₂)₁₄CH₃ (1e)Example 9 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₁₄ CH₃ (1e)

按照实施例5的方法，3.372g(12mmol)Boc-Tyr与2.42g(10mmol)CH₃(C-H₂)₁₅NH₂反应，得到标题化合物3.17g，为无色固体，产率63％.ESI-MS(m/e)：527.7[M+Na]⁺.According to the method of Example 5, 3.372g (12mmol) Boc-Tyr was reacted with 2.42g (10mmol) CH₃ (CH₂ )₁₅ NH₂ to obtain 3.17g of the title compound as a colorless solid with a yield of 63%.ESI- MS(m/e): 527.7[M+Na]⁺ .

实施例10制备Boc-Tyr-NHCH₂(CH₂)₁₆CH₃ (1f)Example 10 Preparation of Boc-Tyr-NHCH₂ (CH₂ )₁₆ CH₃ (1f)

按照实施例5的方法，5.062g(18mmol)Boc-Tyr与4.035g(15mmol)CH₃-(CH₂)₁₇NH₂反应，得到标题化合物7g，为无色固体，产率88％.ESI-MS(m/e)：555.5[M+Na]⁺.According to the method of Example 5, 5.062g (18mmol) Boc-Tyr was reacted with 4.035g (15mmol) CH₃ -(CH₂ )₁₇ NH₂ to obtain the title compound 7g as a colorless solid with a yield of 88%.ESI- MS(m/e): 555.5[M+Na]⁺ .

实施例11制备Boc-Tyr-OCH₂(CH₂)₆CH₃ (1g)Example 11 Preparation of Boc-Tyr-OCH₂ (CH₂ )₆ CH₃ (1g)

将4.125g(15mmol)Boc-Tyr溶于30ml无水四氢呋喃(THF)，冰浴下往里加0.525g(15mmol)N-羟基苯并三唑(HOBt)和3.708g(18mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟，得对应的活泼酯溶液，待用。Dissolve 4.125g (15mmol) Boc-Tyr in 30ml anhydrous tetrahydrofuran (THF), add 0.525g (15mmol) N-hydroxybenzotriazole (HOBt) and 3.708g (18mmol) dicyclohexylcarbonyldi imine (DCC) in anhydrous THF. The reaction mixture was stirred in an ice bath for 30 minutes to obtain the corresponding active ester solution for use.

将2.34g(18mmol)CH₃(CH₂)₇OH溶于20ml无水四氢呋喃(THF)，然后与上面待用的活泼酯溶液混溶。得到的反应混合物室温反应24小时。TLC(展开剂CHCl₃∶CH₃OH＝30∶1)显示Boc-Tyr消失。反应混合物减压浓缩至干，残留物用乙酸乙酯溶解，滤除不溶物。滤液依次用5％碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次；5％硫酸氢钾水溶液洗3次，饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩，柱层析纯化，得1.204g标题化合物，为无色油状物，产率20％.ESI-MS(m/e)：416.7[M+Na]⁺.2.34 g (18 mmol) of CH₃ (CH₂ )₇ OH were dissolved in 20 ml of anhydrous tetrahydrofuran (THF), and then miscible with the active ester solution to be used above. The resulting reaction mixture was reacted at room temperature for 24 hours. TLC (developer CHCl₃ :CH₃ OH=30:1) showed that Boc-Tyr disappeared. The reaction mixture was concentrated to dryness under reduced pressure, the residue was dissolved in ethyl acetate, and the insoluble matter was filtered off. The filtrate was washed 3 times with 5% sodium bicarbonate aqueous solution successively, and saturated sodium chloride aqueous solution was washed 3 times; 5% potassium hydrogensulfate aqueous solution was washed 3 times, saturated sodium chloride aqueous solution was washed 3 times and saturated sodium bicarbonate aqueous solution was washed 3 times, Wash with saturated sodium chloride aqueous solution 3 times. The separated ethyl acetate layer was dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure at 32°C, and purified by column chromatography to obtain 1.204 g of the title compound as a colorless oil, with a yield of 20%. ESI-MS (m /e): 416.7[M+Na]⁺ .

实施例12制备Boc-Tyr-OCH₂(CH₂)₈CH₃ (1h)Example 12 Preparation of Boc-Tyr-OCH₂ (CH₂ )₈ CH₃ (1h)

按照实施例11的方法，3.372g(12mmol)Boc-Tyr与2.775g(15mmol)CH₃-(CH₂)₉OH反应，得到标题化合物2.5g，为无色油状物，产率49％.ESI-MS(m/e)：444.7[M+Na]⁺.According to the method of Example 11, 3.372g (12mmol) Boc-Tyr was reacted with 2.775g (15mmol) CH₃ -(CH₂ )₉ OH to obtain 2.5g of the title compound as a colorless oil with a yield of 49%.ESI -MS(m/e): 444.7[M+Na]⁺ .

实施例13制备Boc-Tyr-OCH₂(CH₂)₁₀CH₃ (1i)Example 13 Preparation of Boc-Tyr-OCH₂ (CH₂ )₁₀ CH₃ (1i)

按照实施例24的方法，2.81g(10mmol)Boc-Tyr与2.232g(12mmol)CH₃-(CH₂)₁₁OH反应，得到标题化合物1.4g，为无色油状物，产率31％.ESI-MS(m/e)：472.8[M+Na]⁺.According to the method of Example 24, 2.81g (10mmol) Boc-Tyr was reacted with 2.232g (12mmol) CH₃ -(CH₂ )₁₁ OH to obtain 1.4g of the title compound as a colorless oil with a yield of 31%.ESI -MS(m/e): 472.8[M+Na]⁺ .

实施例14制备Boc-Tyr-OCH₂(CH₂)₁₂CH₃ (1j)Example 14 Preparation of Boc-Tyr-OCH₂ (CH₂ )₁₂ CH₃ (1j)

按照实施例11的方法，5.058g(19.8mmol)Boc-Tyr与4.922g(23mmol)CH₃-(CH₂)₁₃OH反应，得到标题化合物2.2g，为无色油状物，产率23％.ESI-MS(m/e)：500.7[M+Na]⁺.According to the method of Example 11, 5.058g (19.8mmol) Boc-Tyr was reacted with 4.922g (23mmol) CH₃ -(CH₂ )₁₃ OH to obtain 2.2g of the title compound as a colorless oil with a yield of 23%. ESI-MS(m/e): 500.7[M+Na]⁺ .

实施例15制备Boc-Tyr-OCH₂(CH₂)₁₄CH₃ (1k)Example 15 Preparation of Boc-Tyr-OCH₂ (CH₂ )₁₄ CH₃ (1k)

按照实施例11的方法，5.06g(20mmol)Boc-Tyr与5.808g(24mmol)CH₃-(CH₂)₁₅OH反应，得到标题化合物2.4g，为无色油状物，产率24％.ESI-MS(m/e)：528.8[M+Na]⁺.According to the method of Example 11, 5.06g (20mmol) Boc-Tyr was reacted with 5.808g (24mmol) CH₃ -(CH₂ )₁₅ OH to obtain 2.4g of the title compound as a colorless oil with a yield of 24%.ESI -MS (m/e): 528.8[M+Na]⁺ .

实施例16制备Boc-Tyr-OCH₂(CH₂)₁₆CH₃ (1l)Example 16 Preparation of Boc-Tyr-OCH₂ (CH₂ )₁₆ CH₃ (1l)

按照实施例11的方法，6.744g(24mmol)Boc-Tyr与5.4g(20mmol)CH₃-(CH₂)₁₇OH反应，得到标题化合物2.9g，为无色油状物，产率27％.ESI-MS(m/e)：556.8[M+Na]⁺.According to the method of Example 11, 6.744g (24mmol) Boc-Tyr was reacted with 5.4g (20mmol) CH₃ -(CH₂ )₁₇ OH to obtain 2.9g of the title compound as a colorless oil with a yield of 27%.ESI -MS (m/e): 556.8[M+Na]⁺ .

实施例17制备HCl·Tyr-NHCH₂(CH₂)₆CH₃ (2a)Example 17 Preparation of HCl·Tyr-NHCH₂ (CH₂ )₆ CH₃ (2a)

3.0g(7.65mmol)Boc-Tyr-NHCH₂(CH₂)₆CH₃溶于适量乙酸乙酯，加入约25ml4N无水氯化氢-乙酸乙酯液，室温搅拌3小时，TLC(展开剂石油醚∶丙酮＝5∶1)显示原料点消失。反应混合液在室温下减压浓缩，残留物再用乙酸乙酯溶解并室温下浓缩，如此反复4次；然后向浓缩物中加入乙醚，室温下减压浓缩，如此反复3次，直至除净游离的氯化氢，得标题化合物2.45g，为无色固体，产率97％.ESI-MS(m/e)：293.5[M+H]⁺.3.0g (7.65mmol) Boc-Tyr-NHCH₂ (CH₂ )₆ CH₃ was dissolved in an appropriate amount of ethyl acetate, and about 25ml of 4N anhydrous hydrogen chloride-ethyl acetate solution was added, stirred at room temperature for 3 hours, TLC (petroleum ether: Acetone=5:1) showed disappearance of raw material spots. The reaction mixture was concentrated under reduced pressure at room temperature, and the residue was dissolved in ethyl acetate and concentrated at room temperature, and this was repeated 4 times; then ether was added to the concentrate, and concentrated under reduced pressure at room temperature, and this was repeated 3 times until the Free hydrogen chloride gave 2.45g of the title compound as a colorless solid with a yield of 97%. ESI-MS (m/e): 293.5[M+H]⁺ .

实施例18制备HCl·Tyr-NHCH₂(CH₂)₈CH₃ (2b)Example 18 Preparation of HCl Tyr-NHCH₂ (CH₂ )₈ CH₃ (2b)

按照实施例17的方法，以3.1g(7.38mmol)Boc-Tyr-NHCH₂(CH₂)₈CH₃为原料，得标题化合物2.01g，为无色固体，产率97％.ESI-MS(m/e)：321.5[M+H]⁺.According to the method of Example 17, using 3.1 g (7.38 mmol) of Boc-Tyr-NHCH₂ (CH₂ )₈ CH₃ as raw material, 2.01 g of the title compound was obtained as a colorless solid with a yield of 97%. ESI-MS ( m/e): 321.5[M+H]⁺ .

实施例19制备HCl·Tyr-NH CH₂(CH₂)₁₀CH₃ (2c)Example 19 Preparation of HCl Tyr-NH CH₂ (CH₂ )₁₀ CH₃ (2c)

按照实施例17的方法，以2.75g(5.58mmol)Boc-Tyr-NHCH₂(CH₂)₁₀CH₃为原料，得标题化合物2.05g，为无色固体，产率94％.ESI-MS(m/e)：349.5[M+H]⁺.According to the method of Example 17, using 2.75 g (5.58 mmol) of Boc-Tyr-NHCH₂ (CH₂ )₁₀ CH₃ as raw material, 2.05 g of the title compound was obtained as a colorless solid with a yield of 94%. ESI-MS ( m/e): 349.5[M+H]⁺ .

实施例20制备HCl·Tyr-NHCH₂(CH₂)₁₂CH₃ (2d)Example 20 Preparation of HCl Tyr-NHCH₂ (CH₂ )₁₂ CH₃ (2d)

按照实施例17的方法，以4.0g(8.4mmol)Boc-Tyr-NHCH₂(CH₂)₁₂CH₃为原料，得标题化合物2.98g，为无色固体，产率86％.ESI-MS(m/e)：377.6[M+H]⁺.According to the method of Example 17, using 4.0 g (8.4 mmol) of Boc-Tyr-NHCH₂ (CH₂ )₁₂ CH₃ as raw material, 2.98 g of the title compound was obtained as a colorless solid with a yield of 86%. ESI-MS ( m/e): 377.6[M+H]⁺ .

实施例21制备HCl·Tyr-NHCH₂(CH₂)₁₄CH₃ (2e)Example 21 Preparation of HCl Tyr-NHCH₂ (CH₂ )₁₄ CH₃ (2e)

按照实施例17的方法，以3.17g(6.29mmol)Boc-Tyr-NHCH₂(CH₂)₁₄CH₃为原料，得标题化合物2.67g，为无色固体，产率96％.ESI-MS(m/e)：405.6[M+H]⁺.According to the method of Example 17, using 3.17 g (6.29 mmol) of Boc-Tyr-NHCH₂ (CH₂ )₁₄ CH₃ as raw material, 2.67 g of the title compound was obtained as a colorless solid with a yield of 96%. ESI-MS ( m/e): 405.6[M+H]⁺ .

实施例22制备HCl·Tyr-NHCH₂(CH₂)₁₆CH₃ (2f)Example 22 Preparation of HCl Tyr-NHCH₂ (CH₂ )₁₆ CH₃ (2f)

按照实施例17的方法，以7.0g(13.2mmol)Boc-Tyr-NHCH₂(CH₂)₁₆CH₃为原料，得标题化合物5.5g，为无色固体，产率89％.ESI-MS(m/e)：433.5[M+H]⁺.According to the method of Example 17, using 7.0 g (13.2 mmol) of Boc-Tyr-NHCH₂ (CH₂ )₁₆ CH₃ as raw material, 5.5 g of the title compound was obtained as a colorless solid with a yield of 89%. ESI-MS ( m/e): 433.5[M+H]⁺ .

实施例23制备HCl·Tyr-OCH₂(CH₂)₆CH₃ (2g)Example 23 Preparation of HCl·Tyr-OCH₂ (CH₂ )₆ CH₃ (2g)

按照实施例17的方法，以2.2g(5.6mmol)Boc-Tyr-OCH₂(CH₂)₆CH₃为原料，得标题化合物1.8g，为无色固体，产率98％.ESI-MS(m/e)：294.7[M+H]⁺.According to the method of Example 17, using 2.2 g (5.6 mmol) of Boc-Tyr-OCH₂ (CH₂ )₆ CH₃ as raw material, 1.8 g of the title compound was obtained as a colorless solid with a yield of 98%. ESI-MS ( m/e): 294.7[M+H]⁺ .

实施例24制备HCl·Tyr-OCH₂(CH₂)₈CH₃ (2h)Example 24 Preparation of HCl Tyr-OCH₂ (CH₂ )₈ CH₃ (2h)

按照实施例17的方法，以2.5g(5.94mmol)Boc-Tyr-OCH₂(CH₂)₈CH₃为原料，得标题化合物2.02g，为无色固体，产率95％.ESI-MS(m/e)：322.6[M+H]⁺.According to the method of Example 17, using 2.5 g (5.94 mmol) of Boc-Tyr-OCH₂ (CH₂ )₈ CH₃ as raw material, 2.02 g of the title compound was obtained as a colorless solid with a yield of 95%. ESI-MS ( m/e): 322.6[M+H]⁺ .

实施例25制备HCl·Tyr-OCH₂(CH₂)₁₀CH₃ (2i)Example 25 Preparation of HCl·Tyr-OCH₂ (CH₂ )₁₀ CH₃ (2i)

按照实施例17的方法，以1.9g(4.23mmol)Boc-Tyr-OCH₂(CH₂)₁₀CH₃为原料，得标题化合物1.53g，为无色固体，产率94％.ESI-MS(m/e)：350.5[M+H]⁺.According to the method of Example 17, using 1.9 g (4.23 mmol) of Boc-Tyr-OCH₂ (CH₂ )₁₀ CH₃ as raw material, 1.53 g of the title compound was obtained as a colorless solid with a yield of 94%. ESI-MS ( m/e): 350.5[M+H]⁺ .

实施例26制备HCl·Tyr-OCH₂(CH₂)₁₂CH₃ (2j)Example 26 Preparation of HCl·Tyr-OCH₂ (CH₂ )₁₂ CH₃ (2j)

按照实施例17的方法，以2.2g(4.61mmol)Boc-Tyr-OCH₂(CH₂)₁₂CH₃为原料，得标题化合物1.87g，为无色固体，产率98％.ESI-MS(m/e)：378.6[M+H]⁺.According to the method of Example 17, using 2.2 g (4.61 mmol) of Boc-Tyr-OCH₂ (CH₂ )₁₂ CH₃ as raw material, 1.87 g of the title compound was obtained as a colorless solid with a yield of 98%. ESI-MS ( m/e): 378.6[M+H]⁺ .

实施例27制备HCl·Tyr-OCH₂(CH₂)₁₄CH₃ (2k)Example 27 Preparation of HCl·Tyr-OCH₂ (CH₂ )₁₄ CH₃ (2k)

按照实施例17的方法，以2.4g(4.75mmol)Boc-Tyr-OCH₂(CH₂)₁₄CH₃为原料，得标题化合物2.02g，为无色固体，产率96％.ESI-MS(m/e)：406.6[M+H]⁺.According to the method of Example 17, using 2.4 g (4.75 mmol) of Boc-Tyr-OCH₂ (CH₂ )₁₄ CH₃ as raw material, 2.02 g of the title compound was obtained as a colorless solid with a yield of 96%. ESI-MS ( m/e): 406.6[M+H]⁺ .

实施例28制备HCl·Tyr-OCH₂(CH₂)₁₆CH₃ (2l)Example 28 Preparation of HCl·Tyr-OCH₂ (CH₂ )₁₆ CH₃ (2l)

按照实施例17的方法，以2.9g(5.44mmol)Boc-Tyr-OCH₂(CH₂)₁₆CH₃为原料，得标题化合物2.46g，为无色固体，产率97％.ESI-MS(m/e)：434.5[M+H]⁺.According to the method of Example 17, using 2.9 g (5.44 mmol) of Boc-Tyr-OCH₂ (CH₂ )₁₆ CH₃ as raw material, 2.46 g of the title compound was obtained as a colorless solid with a yield of 97%. ESI-MS ( m/e): 434.5[M+H]⁺ .

实施例29制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OMeExample 29 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OMe

将2.807g(8.8mmol)N^α-Boc-N^G-Arg(NO₂)溶于20ml无水四氢呋喃(THF)，冰浴下往里加1.08g(8mmol)N-羟基苯并三唑(HOBt)和1.978g(9.6mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟，得对应的活泼酯溶液，待用。Dissolve 2.807g (8.8mmol) N^α -Boc-N^G -Arg (NO₂ ) in 20ml anhydrous tetrahydrofuran (THF), add 1.08g (8mmol) N-hydroxybenzotriazole (HOBt) to it under ice bath and 1.978 g (9.6 mmol) of dicyclohexylcarbodiimide (DCC) in anhydrous THF. The reaction mixture was stirred in an ice bath for 30 minutes to obtain the corresponding active ester solution for use.

将1.852g(6.57mmol)HCl·Tyr-OMe溶于30ml无水四氢呋喃(THF)，然后与上面待用的活泼酯溶液混溶，用N-甲基吗啉(NMM)调pH＝8～9。得到的反应混合物室温反应14小时。TLC显示HCl·Tyr-OMe消失。反应混合物减压浓缩至干，残留物用乙酸乙酯溶解，滤除不溶物。滤液依次用5％碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次；5％硫酸氢钾水溶液洗3次，饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩，柱层析纯化，得标题化合物1.7g，为无色固体，产率52％.Mp 72℃，ESI-MS(m/z)：519.6[M+Na]⁺.IR(KBr)：3325.14，2958.48，1657.85，1524.64，1260.73.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.21(s，1H)，8.49(s，1H)，8.12-8.09(d，J＝9Hz，1H)，7.90-7.88(m，1H)，6.70-6.97(d，J＝9Hz，2H)，6.83-6.81(d，J＝6Hz，1H)，6.67-6.64(d，J＝9Hz，3H)，4.43-4.37(Q，J＝6Hz，1H)，3.95-3.93(m，1H)，3.13-3.11(d，J＝6Hz，2H)，2.88-2.81(m，2H)，1.57-1.52(m，2H)，1.45(s，2H)，1.38(s，9H).Dissolve 1.852g (6.57mmol) of HCl Tyr-OMe in 30ml of anhydrous tetrahydrofuran (THF), and then mix with the active ester solution to be used above, adjust the pH to 8-9 with N-methylmorpholine (NMM) . The resulting reaction mixture was reacted at room temperature for 14 hours. TLC showed that HCl·Tyr-OMe disappeared. The reaction mixture was concentrated to dryness under reduced pressure, the residue was dissolved in ethyl acetate, and the insoluble matter was filtered off. The filtrate was washed 3 times with 5% sodium bicarbonate aqueous solution successively, and saturated sodium chloride aqueous solution was washed 3 times; 5% potassium hydrogensulfate aqueous solution was washed 3 times, saturated sodium chloride aqueous solution was washed 3 times and saturated sodium bicarbonate aqueous solution was washed 3 times, Wash with saturated sodium chloride aqueous solution 3 times. The separated ethyl acetate layer was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 32°C, and purified by column chromatography to obtain 1.7 g of the title compound as a colorless solid with a yield of 52%. Mp 72°C, ESI-MS (m/z): 519.6[M+Na]⁺ .IR (KBr): 3325.14, 2958.48, 1657.85, 1524.64, 1260.73.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm=9.21 (s, 1H), 8.49(s, 1H), 8.12-8.09(d, J=9Hz, 1H), 7.90-7.88(m, 1H), 6.70-6.97(d, J=9Hz, 2H), 6.83- 6.81(d, J=6Hz, 1H), 6.67-6.64(d, J=9Hz, 3H), 4.43-4.37(Q, J=6Hz, 1H), 3.95-3.93(m, 1H), 3.13-3.11( d, J=6Hz, 2H), 2.88-2.81(m, 2H), 1.57-1.52(m, 2H), 1.45(s, 2H), 1.38(s, 9H).

实施例30制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₆CH₃ (3a)Example 30 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₆ CH₃ (3a)

将1.914g(6mmol)N^α-Boc-N^G-Arg(NO₂)溶于20ml无水四氢呋喃(THF)，冰浴下往里加0.599g(4.44mmol)N-羟基苯并三唑(HOBt)和1.098g(5.33mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟，得对应的活泼酯溶液，待用。Dissolve 1.914g (6mmol) N^α -Boc-N^G -Arg (NO₂ ) in 20ml anhydrous tetrahydrofuran (THF), add 0.599g (4.44mmol) N-hydroxybenzotriazole (HOBt) to it under ice bath and 1.098 g (5.33 mmol) of dicyclohexylcarbodiimide (DCC) in anhydrous THF. The reaction mixture was stirred in an ice bath for 30 minutes to obtain the corresponding active ester solution for use.

将1.46g(4.44mmol)HCl·Tyr-NHCH₂(CH₂)₆CH₃溶于30ml无水四氢呋喃(THF)，然后与上面待用的活泼酯溶液混溶，用N-甲基吗啉(NMM)调pH＝8～9。得到的反应混合物室温反应14小时。TLC显示HCl·Tyr-NHCH₂(CH₂)₆CH₃消失。反应混合物减压浓缩至干，残留物用乙酸乙酯溶解，滤除不溶物。滤液依次用5％碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次；5％硫酸氢钾水溶液洗3次，饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次，饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩，柱层析纯化，得标题化合物1.5g，为无色固体，产率57％.Mp 87℃，

ESI-MS(m/z)：592.9[M-H]^-.IR(KBr)：3318.41，2930.42，1650.26，1529.87，1263.40，684.95.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.11(s，1H)，8.45(s，1H)，7.78(m，1H)，7.70-7.67(d，J＝9Hz，2H)，6.97-6.94(d，J＝9Hz，2H)，6.63-6.61(d，J＝6Hz，2H)，4.40-4.33(Q，J＝6Hz，1H)，3.86-3.84(m，1H)，3.10-2.90(m，4H)，2.83-2.72(m，2H)，1.54-1.47(m，6H)，1.38(s，9H)，1.23(s，10H)，0.88-0.84(t，J＝6.0Hz，3H).Dissolve 1.46g (4.44mmol) of HCl·Tyr-NHCH₂ (CH₂ )₆ CH₃ in 30ml of anhydrous tetrahydrofuran (THF), and then mix with the active ester solution to be used above, and use N-methylmorpholine ( NMM) adjust pH=8～9. The resulting reaction mixture was reacted at room temperature for 14 hours. TLC showed the disappearance of HCl·Tyr-NHCH₂ (CH₂ )₆ CH₃ . The reaction mixture was concentrated to dryness under reduced pressure, the residue was dissolved in ethyl acetate, and the insoluble matter was filtered off. The filtrate was washed 3 times with 5% sodium bicarbonate aqueous solution successively, and saturated sodium chloride aqueous solution was washed 3 times; 5% potassium hydrogensulfate aqueous solution was washed 3 times, saturated sodium chloride aqueous solution was washed 3 times and saturated sodium bicarbonate aqueous solution was washed 3 times, Wash with saturated sodium chloride aqueous solution 3 times. The separated ethyl acetate layer was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 32°C, and purified by column chromatography to obtain 1.5 g of the title compound as a colorless solid with a yield of 57%. Mp 87°C,

ESI-MS (m/z): 592.9 [MH]^- .IR (KBr): 3318.41, 2930.42, 1650.26, 1529.87, 1263.40, 684.95.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm=9.11 (s, 1H), 8.45 (s, 1H), 7.78 (m, 1H), 7.70-7.67 (d, J=9Hz, 2H), 6.97-6.94 (d, J=9Hz, 2H), 6.63-6.61 ( d, J=6Hz, 2H), 4.40-4.33(Q, J=6Hz, 1H), 3.86-3.84(m, 1H), 3.10-2.90(m, 4H), 2.83-2.72(m, 2H), 1.54 -1.47(m, 6H), 1.38(s, 9H), 1.23(s, 10H), 0.88-0.84(t, J=6.0Hz, 3H).

实施例31制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₈CH₃ (3b)Example 31 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₈ CH₃ (3b)

按照实施例30的方法，3.062g(9.6mmol)N^α-Boc-N^G-Arg(NO₂)与2.56g(7.18mmol)HCl·Tyr-NH-CH₂(CH₂)₈CH₃反应，得标题化合物3.05g，为无色固体，产率68％.Mp 98℃，

ESI-MS(m/z)：644.3[M+Na]⁺.IR(KBr)：3312.67，2934.92，1646.79，1529.90，1262.13，1165.64，683.16.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.15(s，1H)，8.49-8.47(m，1H)，7.91(s，1H)，7.81(m，2H)，7.72-7.69(d，J＝9Hz，1H)，6.97-6.94(d，J＝9Hz，3H)，6.63-6.61(d，J＝6Hz，2H)，4.39-4.34(m，1H)，3.84(s，1H)，3.09(m，2H)，3.03-2.89(m，1H)，2.83-2.69(m，2H)，1.54-1.47(m，2H)，1.38(s，9H)，1.44-1.37(m，4H)，1.23(m，10H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 30, 3.062g (9.6mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 2.56g (7.18mmol) HCl Tyr-NH-CH₂ (CH₂ )₈ CH₃ , 3.05 g of the title compound was obtained as a colorless solid with a yield of 68%. Mp 98°C,

ESI-MS (m/z): 644.3[M+Na]⁺ .IR (KBr): 3312.67, 2934.92, 1646.79, 1529.90, 1262.13, 1165.64, 683.16.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.15(s, 1H), 8.49-8.47(m, 1H), 7.91(s, 1H), 7.81(m, 2H), 7.72-7.69(d, J=9Hz, 1H), 6.97-6.94( d, J=9Hz, 3H), 6.63-6.61(d, J=6Hz, 2H), 4.39-4.34(m, 1H), 3.84(s, 1H), 3.09(m, 2H), 3.03-2.89(m , 1H), 2.83-2.69(m, 2H), 1.54-1.47(m, 2H), 1.38(s, 9H), 1.44-1.37(m, 4H), 1.23(m, 10H), 0.86-0.82(t , J=6.0Hz, 3H).

实施例32制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₁₀CH₃ (3c)Example 32 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₁₀ CH₃ (3c)

按照实施例30的方法，1.914g(6mmol)N^α-Boc-N^G-Arg(NO₂)与2.05g(5mmol)HCl·Tyr-NH-CH₂(CH₂)₁₀CH₃反应，得标题化合物2.2g，为无色固体，产率64％.Mp 104℃，

ESI-MS(m/z)：672.3[M+Na]⁺.IR(KBr)：3319.46，2925.19，2855.17，1638.89，1520.02，1260.59，1166.98，684.44.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.15(s，1H)，8.47(s，1H)，7.91(s，1H)，7.81(s，2H)，7.72-7.70(d，J＝6Hz，1H)，6.96-6.93(d，J＝9Hz，3H)，6.63-6.60(d，J＝9Hz，2H)，4.39(s，1H)，3.84(s，1H)，3.08(m，2H)，3.03-2.94(m，1H)，2.80-2.72(m，1H)，1.37(m，15H)，1.22(s，18H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 30, 1.914g (6mmol) N^α -Boc-^NG -Arg (NO₂ ) was reacted with 2.05g (5 mmol) HCl Tyr-NH-CH₂ (CH₂ )₁₀ CH₃ to obtain the title Compound 2.2g, a colorless solid, yield 64%. Mp 104°C,

ESI-MS (m/z): 672.3[M+Na]⁺ .IR (KBr): 3319.46, 2925.19, 2855.17, 1638.89, 1520.02, 1260.59, 1166.98, 684.44.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.15(s, 1H), 8.47(s, 1H), 7.91(s, 1H), 7.81(s, 2H), 7.72-7.70(d, J=6Hz, 1H), 6.96-6.93( d, J=9Hz, 3H), 6.63-6.60(d, J=9Hz, 2H), 4.39(s, 1H), 3.84(s, 1H), 3.08(m, 2H), 3.03-2.94(m, 1H ), 2.80-2.72(m, 1H), 1.37(m, 15H), 1.22(s, 18H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例33制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₁₂CH₃ (3d)Example 33 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₁₂ CH₃ (3d)

按照实施例30的方法，1.6g(5mmol)N^α-Boc-N^G-Arg(NO₂)与1.504g(3.6mmol)HCl·Tyr-NH-CH₂(CH₁₂CH₃反应，得标题化合物0.81g，为无色固体，产率33％.Mp 105℃，ESI-MS(m/z)：700.3[M+Na]⁺.IR(KBr)：3326.27，2930.89，2342.47，1652.85，1529.62，1263.94，672.84.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.16(s，1H)，8.48(s，1H)，7.94(s，1H)，7.82(m，1H)，7.72-7.70(d，J＝6Hz，1H)，7.00(s，2H)，6.96-6.93(d，J＝9Hz，2H)，6.63-6.60(d，J＝9Hz，2H)，4.37-4.35(m，1H)，3.83(m，1H)，3.08(m，2H)，3.03-2.89(m，3H)，2.86-2.67(m，1H)，1.37(s，9H)，1.22(m，28H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 30, 1.6 g (5 mmol) N^α -Boc-^NG -Arg (NO₂ ) was reacted with 1.504 g (3.6 mmol) HCl Tyr-NH-CH₂ (CH₁₂ CH_{3 )} to obtain the title compound 0.81g, as a colorless solid, yield 33%.Mp 105°C, ESI-MS (m/z): 700.3[M+Na]⁺ .IR (KBr): 3326.27, 2930.89, 2342.47, 1652.85, 1529.62, 1263.94, 672.84.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.16(s, 1H), 8.48(s, 1H), 7.94(s, 1H), 7.82(m, 1H), 7.72-7.70(d, J=6Hz, 1H), 7.00(s, 2H) , 6.96-6.93(d, J=9Hz, 2H), 6.63-6.60(d, J=9Hz, 2H), 4.37-4.35(m, 1H), 3.83(m, 1H), 3.08(m, 2H), 3.03-2.89(m, 3H), 2.86-2.67(m, 1H), 1.37(s, 9H), 1.22(m, 28H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例34制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₁₄CH₃ (3e)Example 34 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₁₄ CH₃ (3e)

按照实施例30的方法，1.914g(6mmol)N^α-Boc-N^G-Arg(NO₂)与2.0g(5mmol)HCl·Tyr-NH-CH₂(CH₂)₁₄CH₃反应，得标题化合物1.96g，为无色固体，产率56％.Mp110℃，

ESI-MS(m/z)：728.5[M+Na]⁺.IR(KBr)：3321.22，2931.61，1638.03，1533.06，1258.94，685.21.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.14(s，1H)，8.48(s，1H)，7.94-7.92(m，1H)，7.84-7.80(d，J＝6Hz，1H)，7.72-7.70(d，J＝6Hz，1H)，7.00-6.98(d，J＝6Hz，1H)，6.96-6.93(d，J＝9Hz，2H)，6.62-6.59(d，J＝9Hz，2H)，4.39-4.33(Q，J＝6Hz 1H)，3.85-3.83(m，1H)，3.08-3.00(m，3H)，2.97-2.70(m，1H)，2.81-2.69(m，2H)，1.58-1.45(m，6H)，1.37(s，9H)，1.22(s，26H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 1.914g (6mmol) of N^α -Boc-^NG -Arg (NO₂ ) was reacted with 2.0g (5mmol) of HCl·Tyr-NH-CH₂ (CH₂ )₁₄ CH₃ to obtain the title Compound 1.96g, a colorless solid, yield 56%.Mp110°C,

ESI-MS (m/z): 728.5[M+Na]⁺ .IR (KBr): 3321.22, 2931.61, 1638.03, 1533.06, 1258.94, 685.21.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.14(s, 1H), 8.48(s, 1H), 7.94-7.92(m, 1H), 7.84-7.80(d, J=6Hz, 1H), 7.72-7.70(d, J=6Hz, 1H), 7.00-6.98(d, J=6Hz, 1H), 6.96-6.93(d, J=9Hz, 2H), 6.62-6.59(d, J=9Hz, 2H), 4.39-4.33(Q, J=6Hz 1H) , 3.85-3.83(m, 1H), 3.08-3.00(m, 3H), 2.97-2.70(m, 1H), 2.81-2.69(m, 2H), 1.58-1.45(m, 6H), 1.37(s, 9H), 1.22(s, 26H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例35制备N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₁₆CH₃ (3f)Example 35 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₁₆ CH₃ (3f)

按照实施例30的方法，1.914g(6mmol)N^α-Boc-N^G-Arg(NO₂)与2.16g(4.6mmol)HCl·Tyr-NH-CH₂(CH₂)₁₆CH₃反应，得标题化合物1.74g，为无色固体，产率52％.Mp 108℃，

ESI-MS(m/z)：756.4[M+Na]⁺.IR(KBr)：3318.16，2934.39，2344.03，1648.16，1529.94，1262.19，678.71.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.14(s，1H)，8.48(s，1H)，7.91(s，1H)，7.82(m，1H)，7.72-7.69(d，J＝9Hz，1H)，7.21-7.19(m，1H)，7.00(m，1H)，6.96-6.94(d，J＝6Hz，2H)，6.63-6.60(d，J＝9Hz，2H)，4.37-4.31(m，1H)，4.14-4.08(Q，1H)，3.18-3.16(m，2H)，3.07-2.98(m，2H)，2.95-2.91(m，1H)，2.84-2.67(m，1H)，1.38(m，15H)，1.22(m，30H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 30, 1.914g (6mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 2.16g (4.6mmol) HCl·Tyr-NH-CH₂ (CH₂ )₁₆ CH₃ to obtain The title compound 1.74g, is a colorless solid, yield 52%.Mp 108°C,

ESI-MS (m/z): 756.4[M+Na]⁺ .IR (KBr): 3318.16, 2934.39, 2344.03, 1648.16, 1529.94, 1262.19, 678.71.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.14(s, 1H), 8.48(s, 1H), 7.91(s, 1H), 7.82(m, 1H), 7.72-7.69(d, J=9Hz, 1H), 7.21-7.19(m, 1H), 7.00(m, 1H), 6.96-6.94(d, J=6Hz, 2H), 6.63-6.60(d, J=9Hz, 2H), 4.37-4.31(m, 1H), 4.14-4.08(Q , 1H), 3.18-3.16(m, 2H), 3.07-2.98(m, 2H), 2.95-2.91(m, 1H), 2.84-2.67(m, 1H), 1.38(m, 15H), 1.22(m , 30H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例36制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₆CH₃ (3g)Example 36 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₆ CH₃ (3g)

按照实施例30的方法，1.914g(6mmol)N^α-Boc-N^G-Arg(NO₂)与1.64g(4.98mmol)HCl·Tyr-O-CH₂(CH₂)₆CH₃反应，得标题化合物1.4g，为无色固体，产率47％.Mp 55℃，

ESI-MS(m/z)：617.5[M+Na]⁺.IR(KBr)：3326.55，2940.62，1652.29，1523.28，1262.36，574.71.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.24(s，1H)，8.13-8.11(d，J＝6Hz，1H)，7.00-6.97(d，J＝9Hz，2H)，6.86-6.83(d，J＝9Hz，1H)，6.66-6.63(d，J＝9Hz，2H)，4.38-4.34(t，J＝6Hz，1H)，3.97-3.93(t，J＝6Hz 3H)，3.11(s，2H)，2.86-2.84(m，2H)，1.62-1.58(m，2H)，1.57-1.45(m，4H)，1.37(s，9H)，1.23(s，10H)，0.88-0.84(t，J＝6.0Hz，3H).According to the method of Example 30, 1.914g (6mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 1.64g (4.98mmol) HCl·Tyr-O-CH₂ (CH₂ )₆ CH₃ to obtain The title compound 1.4g, is a colorless solid, the yield is 47%.Mp 55°C,

ESI-MS (m/z): 617.5[M+Na]⁺ .IR (KBr): 3326.55, 2940.62, 1652.29, 1523.28, 1262.36, 574.71.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.24(s, 1H), 8.13-8.11(d, J=6Hz, 1H), 7.00-6.97(d, J=9Hz, 2H), 6.86-6.83(d, J=9Hz, 1H), 6.66-6.63 (d, J=9Hz, 2H), 4.38-4.34(t, J=6Hz, 1H), 3.97-3.93(t, J=6Hz 3H), 3.11(s, 2H), 2.86-2.84(m, 2H) , 1.62-1.58(m, 2H), 1.57-1.45(m, 4H), 1.37(s, 9H), 1.23(s, 10H), 0.88-0.84(t, J=6.0Hz, 3H).

实施例37制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₈CH₃ (3h)Example 37 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₈ CH₃ (3h)

按照实施例30的方法，2.105g(6.6mmol)N^α-Boc-N^G-Arg(NO₂)与2.2g(6.15mmol)HCl·Tyr-OCH₂(CH₂)₈CH₃反应，得标题化合物2.0g，为微黄色固体，产率49％.Mp 65℃，

ESI-MS(m/z)：645.6[M+Na]⁺.IR(KBr)：3328.67，2934.64，1648.66，1520.60，1263.29，566.41.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.24(s，1H)，8.13-8.10(d，J＝9Hz，1H)，7.00-6.97(d，J＝9Hz，2H)，6.86-6.83(d，J＝9Hz，2H)，6.66-6.63(d，J＝9Hz，2H)，4.38-4.34(t，J＝6Hz，1H)，3.97-3.93(t，J＝6Hz 3H)，3.11(s，2H)，2.86-2.84(m，2H)，1.57-1.46(m，6H)，1.37(s，9H)，1.23(s，14H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 2.105g (6.6mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 2.2g (6.15mmol) HCl·Tyr-OCH₂ (CH₂ )₈ CH₃ to obtain the title Compound 2.0g, as light yellow solid, yield 49%.Mp 65°C,

ESI-MS (m/z): 645.6[M+Na]⁺ .IR (KBr): 3328.67, 2934.64, 1648.66, 1520.60, 1263.29, 566.41.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.24(s, 1H), 8.13-8.10(d, J=9Hz, 1H), 7.00-6.97(d, J=9Hz, 2H), 6.86-6.83(d, J=9Hz, 2H), 6.66-6.63 (d, J=9Hz, 2H), 4.38-4.34(t, J=6Hz, 1H), 3.97-3.93(t, J=6Hz 3H), 3.11(s, 2H), 2.86-2.84(m, 2H) , 1.57-1.46(m, 6H), 1.37(s, 9H), 1.23(s, 14H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例38制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₁₀CH₃ (3i)Example 38 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₁₀ CH₃ (3i)

按照实施例30的方法，1.194g(6mmol)N^α-Boc-N^G-Arg(NO₂)与1.3g(3.37mmol)HCl·Tyr-OCH₂(CH₂)₁₀CH₃反应，得标题化合物1.05g，为无色固体，产率48％.Mp 60℃，

ESI-MS(m/z)：673.8[M+Na]⁺.IR(KBr)：3325.14，2932.24，1650.58，1524.25，1263.84，566.08.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.23(s，1H)，8.12-8.09(d，J＝9Hz，1H)，7.00-6.97(d，J＝9Hz，2H)，6.85-6.82(d，J＝9Hz，2H)，6.66-6.63(d，J＝9Hz，2H)，4.38-4.34(t，J＝6Hz，1H)，3.97-3.93(t，J＝6Hz 3H)，3.11(s，2H)，2.86-2.84(m，2H)，1.57-1.46(m，6H)，1.37(s，9H)，1.23(s，18H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 1.194g (6mmol) N^α -Boc-^NG -Arg (NO₂ ) was reacted with 1.3g (3.37mmol) HCl Tyr-OCH₂ (CH₂ )₁₀ CH₃ to obtain the title compound 1.05g, as a colorless solid, yield 48%.Mp 60°C,

ESI-MS (m/z): 673.8[M+Na]⁺ .IR (KBr): 3325.14, 2932.24, 1650.58, 1524.25, 1263.84, 566.08.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.23(s, 1H), 8.12-8.09(d, J=9Hz, 1H), 7.00-6.97(d, J=9Hz, 2H), 6.85-6.82(d, J=9Hz, 2H), 6.66-6.63 (d, J=9Hz, 2H), 4.38-4.34(t, J=6Hz, 1H), 3.97-3.93(t, J=6Hz 3H), 3.11(s, 2H), 2.86-2.84(m, 2H) , 1.57-1.46(m, 6H), 1.37(s, 9H), 1.23(s, 18H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例39制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₁₂CH₃ (3j)Example 39 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₁₂ CH₃ (3j)

按照实施例30的方法，1.76g(5.52mmol)N^α-Boc-N^G-Arg(NO₂)与2.1g(5.08mmol)HCl·Tyr-OCH₂(CH₂)₁₂CH₃反应，得标题化合物1.04g，产率32％.Mp 61℃，

ESI-MS(m/z)：679.8[M+H]⁺.IR(KBr)：3325.27，2933.53，1652.46，1524.26，1262.52，571.07.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.23(s，1H)，8.51(s，1H)，8.11-8.09(d，J＝6Hz，1H)，6.99-6.96(d，J＝9Hz，2H)，6.85-6.82(d，J＝9Hz，1H)，6.66-6.63(d，J＝9Hz，2H)，4.39-4.35(t，J＝6Hz，1H)，3.97-3.93(t，J＝6Hz，3H)，3.11(s，2H)，2.86-2.84(m，2H)，1.57(m，2H)，1.46(m，4H)，1.37(s，9H)，1.23(s，22H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 1.76g (5.52mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 2.1g (5.08mmol) HCl·Tyr-OCH₂ (CH₂ )₁₂ CH₃ to obtain the title Compound 1.04g, yield 32%. Mp 61°C,

ESI-MS (m/z): 679.8[M+H]⁺ .IR (KBr): 3325.27, 2933.53, 1652.46, 1524.26, 1262.52, 571.07.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.23(s, 1H), 8.51(s, 1H), 8.11-8.09(d, J=6Hz, 1H), 6.99-6.96(d, J=9Hz, 2H), 6.85-6.82(d, J=9Hz , 1H), 6.66-6.63(d, J=9Hz, 2H), 4.39-4.35(t, J=6Hz, 1H), 3.97-3.93(t, J=6Hz, 3H), 3.11(s, 2H), 2.86-2.84(m, 2H), 1.57(m, 2H), 1.46(m, 4H), 1.37(s, 9H), 1.23(s, 22H), 0.87-0.83(t, J=6.0Hz, 3H) .

实施例40制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₁₄CH₃ (3k)Example 40 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₁₄ CH₃ (3k)

按照实施例30的方法，1.837g(5.76mmol)N^α-Boc-N^G-Arg(NO₂)与1.94g(4.39mmol)HCl·Tyr-OCH₂(CH₂)₁₄CH₃反应，得标题化合物1.8g，为无色固体，产率58％.Mp 64.7℃，

ESI-MS(m/z)：707.9[M+H]⁺.IR(KBr)：3322.96，2933.28，2855.09，1659.90，1519.15，1264.00，569.82.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.22(s，1H)，8.51(s，1H)，8.11-8.08(d，J＝9Hz，1H)，6.99-6.96(d，J＝9Hz，2H)，6.85-6.82(d，J＝9Hz，1H)，6.66-6.63(d，J＝9Hz，2H)，4.39-4.35(m，1H)，3.97-3.93(t，J＝6Hz，3H)，3.11(s，2H)，2.86-2.85(m，2H)，1.58(m，2H)，1.46(m，4H)，1.38(s，9H)，1.23(s，26H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 1.837g (5.76mmol) N^α -Boc-^NG -Arg(NO₂ ) was reacted with 1.94g (4.39mmol) HCl·Tyr-OCH₂ (CH₂ )₁₄ CH₃ to obtain the title Compound 1.8g, a colorless solid, yield 58%. Mp 64.7°C,

ESI-MS (m/z): 707.9[M+H]⁺ .IR (KBr): 3322.96, 2933.28, 2855.09, 1659.90, 1519.15, 1264.00, 569.82.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.22(s, 1H), 8.51(s, 1H), 8.11-8.08(d, J=9Hz, 1H), 6.99-6.96(d, J=9Hz, 2H), 6.85-6.82(d, J =9Hz, 1H), 6.66-6.63(d, J=9Hz, 2H), 4.39-4.35(m, 1H), 3.97-3.93(t, J=6Hz, 3H), 3.11(s, 2H), 2.86- 2.85(m, 2H), 1.58(m, 2H), 1.46(m, 4H), 1.38(s, 9H), 1.23(s, 26H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例41制备N^α-Boc-N^G-Arg(NO₂)-Tyr-OCH₂(CH₂)₁₆CH₃ (3l)Example 41 Preparation of N^α -Boc-N^G -Arg(NO₂ )-Tyr-OCH₂ (CH₂ )₁₆ CH₃ (3l)

按照实施例30的方法，2.1g(6.58mmol)N^α-Boc-N^G-Arg(NO₂)与2.55g(5.44mmol)HCl·Tyr-OCH₂(CH₂)₁₆CH₃反应，得标题化合物1.9g，为无色固体，产率48％.Mp 72℃，

ESI-MS(m/z)：736.3[M+H]⁺.IR(KBr)：3323.01，2925.30，2854.27，1655.84，1518.10，1262.25，1126.54，567.18.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.23(s，1H)，8.51(s，1H)，8.12-8.09(d，J＝9Hz，1H)，6.99-6.96(d，J＝9Hz，2H)，6.86-6.83(d，J＝9Hz，1H)，6.66-6.63(d，J＝9Hz，2H)，4.39-4.35(t，J＝6Hz，1H)，3.97-3.93(t，J＝6Hz，3H)，3.11(s，2H)，2.98-2.89(m，2H)，1.58(m，2H)，1.45(m，4H)，1.37(s，9H)，1.23(m，30H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 30, 2.1g (6.58mmol) of N^α -Boc-^NG -Arg(NO₂ ) was reacted with 2.55g (5.44mmol) of HCl·Tyr-OCH₂ (CH₂ )₁₆ CH₃ to obtain the title Compound 1.9g, a colorless solid, yield 48%.Mp 72°C,

ESI-MS (m/z): 736.3[M+H]⁺ .IR (KBr): 3323.01, 2925.30, 2854.27, 1655.84, 1518.10, 1262.25, 1126.54, 567.18.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.23(s, 1H), 8.51(s, 1H), 8.12-8.09(d, J=9Hz, 1H), 6.99-6.96(d, J=9Hz, 2H), 6.86-6.83(d , J=9Hz, 1H), 6.66-6.63(d, J=9Hz, 2H), 4.39-4.35(t, J=6Hz, 1H), 3.97-3.93(t, J=6Hz, 3H), 3.11(s , 2H), 2.98-2.89(m, 2H), 1.58(m, 2H), 1.45(m, 4H), 1.37(s, 9H), 1.23(m, 30H), 0.87-0.83(t, J=6.0 Hz, 3H).

实施例42制备Arg-TyrExample 42 Preparation of Arg-Tyr

取1.8g(3.63mmol)N^α-Boc-N^G-Arg(NO₂)-OMe于反应瓶中，加入适量甲醇溶解，冰浴搅拌条件下缓慢加入1N NaOH水溶液至pH＝12，冰浴下搅拌反应2h。TLC监测Boc-Arg(NO₂)-Tyr-OMe消失后，用饱和KHSO₄调节反应液pH为7，蒸去甲醇，再将反应液pH调节为2，用乙酸乙酯萃洗反应液三次，得到的乙酸乙酯层用饱和NaCl洗涤三次，随后用无水硫酸钠干燥过夜。滤除硫酸钠，旋除乙酸乙酯得Boc-Arg(NO₂)-Tyr 1.2g，为无色固体。Take 1.8g (3.63mmol) N^α -Boc-N^G -Arg(NO₂ )-OMe in a reaction flask, add an appropriate amount of methanol to dissolve, slowly add 1N NaOH aqueous solution to pH = 12 under ice bath stirring condition, and The reaction was stirred for 2h. After the disappearance of Boc-Arg(NO₂ )-Tyr-OMe was monitored by TLC, the pH of the reaction solution was adjusted to 7 with saturated KHSO₄ , methanol was evaporated, and the pH of the reaction solution was adjusted to 2, and the reaction solution was extracted and washed three times with ethyl acetate. The obtained ethyl acetate layer was washed three times with saturated NaCl, then dried over anhydrous sodium sulfate overnight. Sodium sulfate was filtered off, and ethyl acetate was spin-off to obtain 1.2 g of Boc-Arg(NO₂ )-Tyr as a colorless solid.

将800mg(1.66mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr用10ml CH₃OH溶解，再加入240mg Pd/C(5％)，减压排出反应瓶中的空气，通入氢气置换，反复置换三次后，通氢气室温搅拌反应，直至TLC显示Boc-Arg(NO₂)-Tyr消失。停止反应，滤除Pd/C，滤液减压浓缩至干，得Boc-Arg-Tyr 704mg，为无色固体。Dissolve 800mg (1.66mmol) N^α -Boc-NG^-Arg (NO₂ )-Tyr in 10ml CH₃ OH, then add 240mg Pd/C (5%), depressurize the air in the reaction flask, and inject Hydrogen replacement was repeated three times, and the reaction was stirred at room temperature with hydrogen until TLC showed that Boc-Arg(NO₂ )-Tyr disappeared. The reaction was stopped, Pd/C was filtered off, and the filtrate was concentrated to dryness under reduced pressure to obtain 704 mg of Boc-Arg-Tyr as a colorless solid.

取704mg(1.61mmol)N^α-Boc-N^G-Arg-Tyr溶于适量乙酸乙酯，加入约20ml 4N盐酸乙酸乙酯液，室温搅拌3小时，TLC显示原料点消失。反应混合液在室温下减压浓缩，残留物再用乙酸乙酯溶解并室温下浓缩，如此反复5次；然后向浓缩物中加入乙醚，室温下减压浓缩，如此反复5次，直至除净游离的氯化氢，得标题化合物480mg，为白色固体，产率59％.Mp 123-124℃，

ESI-MS(m/z)：338.2[M+H]⁺.IR(KBr)：3174.85，1665.97，1513.64，1233.75，826.70，565.01.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.38(s，1H)，9.17-9.15(m，1H)，8.98-8.95(m，1H)，8.42-8.38(m，1H)，7.95-7.91(m，1H)，7.58-7.53(m，1H)，7.42-7.39(m，1H)，7.25-7.22(m，1H)，7.09-7.06(d，J＝9Hz，2H)，6.69-6.67(d，J＝6Hz，2H)，4.39-4.35(dd，J＝3Hz，J＝9Hz，1H)，3.88-3.86(m，1H)，3.16(m，2H)，3.05-2.84(m，2H)，1.77-1.76(m，2H)，1.56(m，2H).Take 704 mg (1.61 mmol) of N^α -Boc-N^G -Arg-Tyr and dissolve in an appropriate amount of ethyl acetate, add about 20 ml of 4N hydrochloric acid ethyl acetate solution, stir at room temperature for 3 hours, TLC shows that the raw material point disappears. The reaction mixture was concentrated under reduced pressure at room temperature, and the residue was dissolved in ethyl acetate and concentrated at room temperature, and this was repeated 5 times; then ether was added to the concentrate, and concentrated under reduced pressure at room temperature, and this was repeated 5 times until the Free hydrogen chloride gave 480 mg of the title compound as a white solid with a yield of 59%. Mp 123-124°C,

ESI-MS (m/z): 338.2[M+H]⁺ .IR (KBr): 3174.85, 1665.97, 1513.64, 1233.75, 826.70, 565.01.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.38(s, 1H), 9.17-9.15(m, 1H), 8.98-8.95(m, 1H), 8.42-8.38(m, 1H), 7.95-7.91(m, 1H), 7.58-7.53(m, 1H), 7.42-7.39(m, 1H), 7.25-7.22(m, 1H), 7.09-7.06(d, J=9Hz, 2H), 6.69-6.67(d, J=6Hz, 2H), 4.39-4.35 (dd, J=3Hz, J=9Hz, 1H), 3.88-3.86(m, 1H), 3.16(m, 2H), 3.05-2.84(m, 2H), 1.77-1.76(m, 2H), 1.56( m, 2H).

实施例43制备Arg-Tyr-NHCH₂(CH₂)₆CH₃ (5a)Example 43 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₆ CH₃ (5a)

取430mg(0.73mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NHCH₂(CH₂)₆CH₃溶于适量乙酸乙酯，加入约15ml 4N盐酸乙酸乙酯液，室温搅拌3小时，TLC显示原料点消失。反应混合液在室温下减压浓缩，残留物再用乙酸乙酯溶解并室温下浓缩，如此反复5次；然后向浓缩物中加入乙醚，室温下减压浓缩，如此反复5次，直至除净游离的氯化氢，得标题化合物320mg，为无色固体90％.Mp 126-127℃，

ESI-MS(m/z)：449.4[M+H]⁺.IR(KBr)：3182.43，2936.06，1656.07，1514.07，1239.21，567.11.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.34(s，1H)，8.80-8.77(d，J＝9Hz，1H)，8.44-8.43(d，J＝3Hz，1H)，8.39-8.38(m，1H)，8.10(s，1H)，7.92(s，1H)，7.62-7.59(m，1H)，7.51-7.40(m，1H)，7.29-7.23(m，1H)，7.07-7.04(d，J＝9Hz，2H)，6.68-6.65(d，J＝9Hz，2H)，4.37-4.36(m，1H)，3.82(s，1H)，3.13-3.02(m，3H)，2.93-2.88(m，1H)，2.83-2.72(m，2H)，1.78-1.76(m，2H)，1.54-1.51(m，2H)，1.31-1.27(m，2H)，1.22(s，9H)，0.87-0.83(t，J＝6.0Hz，3H).Take 430mg (0.73mmol) of N^α -Boc-N^G -Arg(NO₂ )-Tyr-NHCH₂ (CH₂ )₆ CH₃ dissolved in an appropriate amount of ethyl acetate, add about 15ml of 4N hydrochloric acid ethyl acetate solution, and stir at room temperature After 3 hours, TLC showed that the starting material spot disappeared. The reaction mixture was concentrated under reduced pressure at room temperature, and the residue was dissolved in ethyl acetate and concentrated at room temperature, and this was repeated 5 times; then ether was added to the concentrate, and concentrated under reduced pressure at room temperature, and this was repeated 5 times until the Free hydrogen chloride gave 320 mg of the title compound as a colorless solid 90%. Mp 126-127°C,

ESI-MS (m/z): 449.4[M+H]⁺ .IR (KBr): 3182.43, 2936.06, 1656.07, 1514.07, 1239.21, 567.11.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.34(s, 1H), 8.80-8.77(d, J=9Hz, 1H), 8.44-8.43(d, J=3Hz, 1H), 8.39-8.38(m, 1H), 8.10(s, 1H), 7.92(s, 1H), 7.62-7.59(m, 1H), 7.51-7.40(m, 1H), 7.29-7.23(m, 1H), 7.07-7.04(d, J=9Hz, 2H), 6.68-6.65 (d, J=9Hz, 2H), 4.37-4.36(m, 1H), 3.82(s, 1H), 3.13-3.02(m, 3H), 2.93-2.88(m, 1H), 2.83-2.72(m, 2H), 1.78-1.76(m, 2H), 1.54-1.51(m, 2H), 1.31-1.27(m, 2H), 1.22(s, 9H), 0.87-0.83(t, J=6.0Hz, 3H) .

实施例44制备Arg-Tyr-NHCH₂(CH₂)₈CH₃ (5b)Example 44 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₈ CH₃ (5b)

按照实施例43的方法，以800mg(1.29mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NH-CH₂(CH₂)₈CH₃为原料，得标题化合物620mg，为无色固体，产率94％.Mp 127-128℃，

ESI-MS(m/z)：477.5[M+H]⁺.IR(KBr)：3101.92，2932.95，1657.55，1515.14，1357.76，1239.89，564.88.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.34(s，1H)，8.79-8.76(d，J＝9Hz，1H)，8.43-8.41(d，J＝6Hz，1H)，8.38-8.36(m，1H)，8.12-8.08(t，J＝6Hz，1H)，7.91(m，1H)，7.51(m，2H)，7.07-7.04(d，J＝9Hz，2H)，6.68-6.65(d，J＝9Hz，2H)，4.40-4.34(m，1H)，3.82-3.81(m，1H)，3.13-3.09(m，2H)，3.07-3.02(m，1H)，2.94-2.87(m，1H)，2.83-2.71(m，2H)，1.78-1.73(m，2H)，1.54-1.49(m，2H)，1.32-1.28(m，2H)，1.27(s，10H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 800 mg (1.29 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-NH-CH₂ (CH₂ )₈ CH₃ as raw material, 620 mg of the title compound was obtained as a free Color solid, yield 94%. Mp 127-128°C,

ESI-MS (m/z): 477.5[M+H]⁺ .IR (KBr): 3101.92, 2932.95, 1657.55, 1515.14, 1357.76, 1239.89, 564.88.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.34(s, 1H), 8.79-8.76(d, J=9Hz, 1H), 8.43-8.41(d, J=6Hz, 1H), 8.38-8.36(m, 1H), 8.12-8.08(t , J=6Hz, 1H), 7.91(m, 1H), 7.51(m, 2H), 7.07-7.04(d, J=9Hz, 2H), 6.68-6.65(d, J=9Hz, 2H), 4.40- 4.34(m, 1H), 3.82-3.81(m, 1H), 3.13-3.09(m, 2H), 3.07-3.02(m, 1H), 2.94-2.87(m, 1H), 2.83-2.71(m, 2H ), 1.78-1.73(m, 2H), 1.54-1.49(m, 2H), 1.32-1.28(m, 2H), 1.27(s, 10H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例45制备Arg-Tyr-NHCH₂(CH₂)₁₀CH₃ (5c)Example 45 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₁₀ CH₃ (5c)

按照实施例43的方法，以700mg(1.08mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NH-CH₂(CH₂)₁₀CH₃为原料，得标题化合物520mg，为无色固体，产率89％.Mp 132-133℃，

ESI-MS(m/z)：505.5[M+Na]⁺.IR(KBr)：3186.92，2931.65，1656.99，1535.35，1472.93，1241.65，821.43，569.81.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.34(s，1H)，8.80-8.77(d，J＝9Hz，1H)，8.44-8.42(d，J＝6Hz，1H)8.39-8.38(m，1H)，8.11(m，1H)，7.92(s，1H)，7.50(m，1H)，7.54-7.42(m，1H)，7.29-7.25(m，1H)，7.07-7.04(d，J＝9Hz，2H)，6.68-6.65(d，J＝9Hz，2H)，4.38-4.34(m，1H)，3.82(s，1H)，3.13-3.03(m，3H)，2.92-2.86(Q，J＝6Hz，1H)，2.83-2.72(m，2H)，1.78-1.76(m，2H)，1.51(m，2H)，1.31-1.27(m，2H)，1.22(s，9H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 43, using 700 mg (1.08 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-NH-CH₂ (CH₂ )₁₀ CH₃ as raw material, 520 mg of the title compound was obtained as a free Color solid, yield 89%. Mp 132-133°C,

ESI-MS (m/z): 505.5[M+Na]⁺ .IR (KBr): 3186.92, 2931.65, 1656.99, 1535.35, 1472.93, 1241.65, 821.43, 569.81.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.34(s, 1H), 8.80-8.77(d, J=9Hz, 1H), 8.44-8.42(d, J=6Hz, 1H), 8.39-8.38(m, 1H), 8.11(m, 1H), 7.92(s, 1H), 7.50(m, 1H), 7.54-7.42(m, 1H), 7.29-7.25(m, 1H), 7.07-7.04(d, J=9Hz, 2H), 6.68- 6.65(d, J=9Hz, 2H), 4.38-4.34(m, 1H), 3.82(s, 1H), 3.13-3.03(m, 3H), 2.92-2.86(Q, J=6Hz, 1H), 2.83 -2.72(m, 2H), 1.78-1.76(m, 2H), 1.51(m, 2H), 1.31-1.27(m, 2H), 1.22(s, 9H), 0.87-0.83(t, J=6.0Hz , 3H).

实施例46制备Arg-Tyr-NHCH₂(CH₂)₁₂CH₃ (5d)Example 46 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₁₂ CH₃ (5d)

按照实施例43的方法，以540mg(0.8mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NH-CH₂(CH₂)₁₂CH₃为原料，得标题化合物340mg，为无色固体，产率75％.Mp 138-139℃，

ESI-MS(m/z)：533.5[M+H]⁺.IR(KBr)：3254.80，2930.14，1657.35，1532.19，1240.84，686.11.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.36(s，1H)，8.78-8.76(d，J＝6Hz，1H)，8.53-8.50(m，1H)，8.41-8.37(m，1H)，8.10(m，1H)，7.91(s，1H)，7.44-7.14(m，1H)，7.07-7.04(d，J＝9Hz，2H)，7.02(s，1H)，6.68-6.65(d，J＝9Hz，2H)，4.38-4.36(m，1H)，3.15(m，2H)，3.13-3.11(m，1H)，2.95-2.88(m，1H)，2.86-2.72(m，2H)，1.78-1.76(m，2H)，1.51(m，2H)，1.33-1.29(m，2H)，1.23(s，22H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 540 mg (0.8 mmol) of N^α -Boc-^NG -Arg(NO₂ )-Tyr-NH-CH₂ (CH₂ )₁₂ CH₃ as raw material, 340 mg of the title compound was obtained as a free Color solid, yield 75%. Mp 138-139°C,

ESI-MS (m/z): 533.5[M+H]⁺ .IR (KBr): 3254.80, 2930.14, 1657.35, 1532.19, 1240.84, 686.11.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.36(s, 1H), 8.78-8.76(d, J=6Hz, 1H), 8.53-8.50(m, 1H), 8.41-8.37(m, 1H), 8.10(m, 1H), 7.91(s, 1H), 7.44-7.14(m, 1H), 7.07-7.04(d, J=9Hz, 2H), 7.02(s, 1H), 6.68-6.65(d, J=9Hz, 2H), 4.38-4.36(m , 1H), 3.15(m, 2H), 3.13-3.11(m, 1H), 2.95-2.88(m, 1H), 2.86-2.72(m, 2H), 1.78-1.76(m, 2H), 1.51(m , 2H), 1.33-1.29(m, 2H), 1.23(s, 22H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例47制备Arg-Tyr-NHCH₂(CH₂)₁₄CH₃(5e)Example 47 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₁₄ CH₃ (5e)

按照实施例43的方法，以400mg(0.57mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NH-CH₂(CH₂)₁₄CH₃为原料，得标题化合物300mg，为无色固体，产率88％.Mp 141-142℃，ESI-MS(m/z)：561.6[M+H]⁺.IR(KBr)：3171.12，2928.33，1656.44，1514.55，1410.90，1241.34，821.01，572.80.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.31(s，1H)，8.78-8.76(d，J＝6Hz，1H)，8.43-8.41(d，J＝6Hz，1H)，8.38-8.37(d，J＝3Hz，1H)，8.12-8.08(t，J＝6Hz，1H)，7.91(s，1H)，7.63-7.57(t，J＝9Hz，1H)，7.46-7.40(t，J＝9Hz，1H)，7.26-7.20(t，J＝9Hz，1H)，7.07-7.04(d，J＝9Hz，2H)，6.68-6.65(d，J＝9Hz，2H)，4.39-4.34(dd，J＝6Hz，J＝9Hz，1H)，3.82-3.81(m，1H)，3.13-3.03(m，3H)，2.94-2.86(m，1H)，2.83-2.71(m，2H)，1.78-1.73(m，2H)，1.54-1.49(m，2H)，1.30-1.29(m，2H)，1.22(s，9H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 400 mg (0.57 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-NH-CH₂ (CH₂ )₁₄ CH₃ as raw material, 300 mg of the title compound was obtained as a free Color solid, yield 88%. Mp 141-142°C, ESI-MS (m/z): 561.6[M+H]⁺ .IR (KBr): 3171.12, 2928.33, 1656.44, 1514.55, 1410.90, 1241.34, 821.01, 572.80.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.31(s, 1H), 8.78-8.76(d, J=6Hz, 1H), 8.43-8.41(d, J=6Hz, 1H), 8.38-8.37(d, J=3Hz, 1H) , 8.12-8.08(t, J=6Hz, 1H), 7.91(s, 1H), 7.63-7.57(t, J=9Hz, 1H), 7.46-7.40(t, J=9Hz, 1H), 7.26-7.20 (t, J=9Hz, 1H), 7.07-7.04(d, J=9Hz, 2H), 6.68-6.65(d, J=9Hz, 2H), 4.39-4.34(dd, J=6Hz, J=9Hz, 1H), 3.82-3.81(m, 1H), 3.13-3.03(m, 3H), 2.94-2.86(m, 1H), 2.83-2.71(m, 2H), 1.78-1.73(m, 2H), 1.54- 1.49(m, 2H), 1.30-1.29(m, 2H), 1.22(s, 9H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例48制备Arg-Tyr-NHCH₂(CH₂)₁₆CH₃ (5f)Example 48 Preparation of Arg-Tyr-NHCH₂ (CH₂ )₁₆ CH₃ (5f)

按照实施例43的方法，以500mg(0.69mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-NH-CH₂(CH₂)₁₆CH₃为原料，得标题化合物340mg，为无色固体，产率79％.Mp 143-144℃，

ESI-MS(m/z)：589.7[M+H]⁺.IR(KBr)：3303.15，2926.49，1653.90，1536.20，1237.58，820.90，687.79，558.56.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝8.78-8.75(d，J＝9Hz，1H)，8.44-8.42(d，J＝6Hz，1H)，8.39-8.35(m，2H)，8.10-8.06(t，J＝6Hz，1H)，7.93(m，1H)，7.53(m，1H)，7.45-7.25(m，1H)，7.07-7.04(d，J＝9Hz，2H)，6.68-6.65(d，J＝9Hz，2H)，4.40-4.34(m，1H)，3.82-3.81(m，1H)，3.13(m，2H)，3.07-3.03(m，1H)，2.94-2.91(m，1H)，2.86-2.77(m，2H)，1.79-1.74(m，2H)，1.58-1.49(m，2H)，1.31-1.27(m，2H)，1.22(s，30H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 500 mg (0.69 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-NH-CH₂ (CH₂ )₁₆ CH₃ as raw material, 340 mg of the title compound was obtained as a free Color solid, yield 79%. Mp 143-144°C,

ESI-MS (m/z): 589.7[M+H]⁺ .IR (KBr): 3303.15, 2926.49, 1653.90, 1536.20, 1237.58, 820.90, 687.79, 558.56.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=8.78-8.75(d, J=9Hz, 1H), 8.44-8.42(d, J=6Hz, 1H), 8.39-8.35(m, 2H), 8.10-8.06(t, J=6Hz, 1H), 7.93(m, 1H), 7.53(m, 1H), 7.45-7.25(m, 1H), 7.07-7.04(d, J=9Hz, 2H), 6.68-6.65(d, J=9Hz, 2H ), 4.40-4.34(m, 1H), 3.82-3.81(m, 1H), 3.13(m, 2H), 3.07-3.03(m, 1H), 2.94-2.91(m, 1H), 2.86-2.77(m , 2H), 1.79-1.74(m, 2H), 1.58-1.49(m, 2H), 1.31-1.27(m, 2H), 1.22(s, 30H), 0.86-0.82(t, J=6.0Hz, 3H ).

实施例49制备Arg-Tyr-OCH₂(CH₂)₆CH₃(5g)Example 49 Preparation of Arg-Tyr-OCH₂ (CH₂ )₆ CH₃ (5g)

按照实施例43的方法，以750mg(1.26mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₆CH₃为原料，得标题化合物510mg，为无色固体，产率83％.Mp 109-110℃，

ESI-MS(m/z)：450.4[M+H]⁺.IR(KBr)：3194.15，2936.20，1660.13，1524.89，1358.14，1231.17，833.14，661.54.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.41(s，1H)，9.19-9.17(d，J＝6Hz，2H)，8.46-8.44(d，J＝6Hz，1H)，8.41-8.39(m，2H)，8.00(s，1H)，7.81(s，1H)，7.07-7.05(d，J＝6Hz，2H)，6.70-6.67(d，J＝9Hz，2H)，4.42-4.34(m，1H)，4.01-3.96(m，2H)，3.94-3.89(m，1H)，3.18-3.14(t，J＝6Hz，2H)，2.92-2.89(d，J＝9Hz，2H)，1.80-1.74(m，2H)，1.60-1.54(m，2H)，1.48-1.44(m，2H)，1.23(s，10H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 43, using 750 mg (1.26 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₆ CH₃ as raw material, 510 mg of the title compound was obtained as a free Color solid, yield 83%. Mp 109-110°C,

ESI-MS (m/z): 450.4[M+H]⁺ .IR (KBr): 3194.15, 2936.20, 1660.13, 1524.89, 1358.14, 1231.17, 833.14, 661.54.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.41(s, 1H), 9.19-9.17(d, J=6Hz, 2H), 8.46-8.44(d, J=6Hz, 1H), 8.41-8.39(m, 2H), 8.00(s , 1H), 7.81(s, 1H), 7.07-7.05(d, J=6Hz, 2H), 6.70-6.67(d, J=9Hz, 2H), 4.42-4.34(m, 1H), 4.01-3.96( m, 2H), 3.94-3.89(m, 1H), 3.18-3.14(t, J=6Hz, 2H), 2.92-2.89(d, J=9Hz, 2H), 1.80-1.74(m, 2H), 1.60 -1.54(m, 2H), 1.48-1.44(m, 2H), 1.23(s, 10H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例50制备Arg-Tyr-OCH₂(CH₂)₈CH₃ (5h)Example 50 Preparation of Arg-Tyr-OCH₂ (CH₂ )₈ CH₃ (5h)

按照实施例43的方法，以800mg(1.29mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₈CH₃为原料，得标题化合物570mg，为无色固体，产率86％.Mp 112-113℃，

ESI-MS(m/z)：478.5[M+H]⁺.IR(KBr)：3340.48，3191.91，2843.74，1669.32，1359.81，1238.80，834.75，671.82.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.22-9.20(d，J＝6Hz，2H)，8.49-8.47(d，J＝6Hz，1H)，8.43-8.39(m，2H)，8.00(s，1H)，7.54(s，1H)，7.07-7.04(d，J＝9Hz，2H)，6.71-6.68(d，J＝9Hz，2H)，4.42-4.35(m，1H)，4.03-3.91(m，3H)，3.17(s，2H)，2.92-2.89(d，J＝9Hz，2H)，1.80-1.76(m，2H)，1.60-1.55(m，2H)，1.48-1.46(m，2H)，1.23(s，14H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 800 mg (1.29 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₈ CH₃ as raw material, 570 mg of the title compound was obtained as a free Color solid, yield 86%. Mp 112-113°C,

ESI-MS (m/z): 478.5[M+H]⁺ .IR (KBr): 3340.48, 3191.91, 2843.74, 1669.32, 1359.81, 1238.80, 834.75, 671.82.¹ H-NMR (300MHz, DMSO-d₆ ) : δ/ppm=9.22-9.20(d, J=6Hz, 2H), 8.49-8.47(d, J=6Hz, 1H), 8.43-8.39(m, 2H), 8.00(s, 1H), 7.54(s , 1H), 7.07-7.04(d, J=9Hz, 2H), 6.71-6.68(d, J=9Hz, 2H), 4.42-4.35(m, 1H), 4.03-3.91(m, 3H), 3.17( s, 2H), 2.92-2.89(d, J=9Hz, 2H), 1.80-1.76(m, 2H), 1.60-1.55(m, 2H), 1.48-1.46(m, 2H), 1.23(s, 14H ), 0.86-0.82(t, J=6.0Hz, 3H).

实施例51制备Arg-Tyr-OCH₂(CH₂)₁₀CH₃ (5i)Example 51 Preparation of Arg-Tyr-OCH₂ (CH₂ )₁₀ CH₃ (5i)

按照实施例43的方法，以500mg(0.77mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₁₀CH₃为原料，得标题化合物320mg，为无色固体，产率77％.Mp 116-117℃，ESI-MS(m/z)：506.4[M+H]⁺.IR(KBr)：3401.62，3135.64，1660.13，1527.30，1222.44，826.64，565.08.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.38(s，1H)，9.19-9.160(d，J＝9Hz，2H)，8.46-8.44(d，J＝6Hz，1H)，8.41-8.39(m，2H)，7.97(m，1H)，7.61-7.44(m，2H)，7.07-7.04(d，J＝9Hz，2H)，6.70-6.67(d，J＝9Hz，2H)，4.42-4.37(m，1H)，4.08-4.02(m，2H)，3.98-3.89(m，2H)，3.18-3.14(t，J＝6Hz，2H)，2.92-2.89(d，J＝9Hz，2H)，1.79-1.75(m，2H)，1.59-1.54(m，2H)，1.51-1.46(m，2H)，1.23(s，18H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 43, using 500 mg (0.77 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₁₀ CH₃ as raw material, 320 mg of the title compound was obtained as a free Color solid, yield 77%. Mp 116-117°C, ESI-MS (m/z): 506.4[M+H]⁺ .IR (KBr): 3401.62, 3135.64, 1660.13, 1527.30, 1222.44, 826.64, 565.08.¹ H-NMR (300MHz, DMSO-d₆ ): δ /ppm=9.38(s, 1H), 9.19-9.160(d, J=9Hz, 2H), 8.46-8.44(d, J=6Hz, 1H), 8.41-8.39(m, 2H), 7.97(m, 1H ), 7.61-7.44(m, 2H), 7.07-7.04(d, J=9Hz, 2H), 6.70-6.67(d, J=9Hz, 2H), 4.42-4.37(m, 1H), 4.08-4.02( m, 2H), 3.98-3.89(m, 2H), 3.18-3.14(t, J=6Hz, 2H), 2.92-2.89(d, J=9Hz, 2H), 1.79-1.75(m, 2H), 1.59 -1.54(m, 2H), 1.51-1.46(m, 2H), 1.23(s, 18H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例52制备Arg-Tyr-OCH₂(CH₂)₁₂CH₃ (5j)Example 52 Preparation of Arg-Tyr-OCH₂ (CH₂ )₁₂ CH₃ (5j)

按照实施例43的方法，以580mg(0.86mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₁₂CH₃为原料，得标题化合物430mg，为无色固体，产率88％.Mp 139-140℃，ESI-MS(m/z)：534.4[M+H]⁺.IR(KBr)：3336.49，3171.37，2932.91，1658.63，1504.92，1218.19.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.34(s，1H)，9.20-9.18(d，J＝6Hz，1H)，8.38(s，3H)，8.01-7.97(t，J＝6Hz J＝6Hz，1H)，7.80(m，1H)，7.53(m，1H)，7.07-7.05(d，J＝6Hz，2H)，6.70-6.67(d，J＝9Hz，2H)，4.42-4.36(Q，J＝6Hz 1H)，4.04-3.97(m，2H)，3.94-3.88(m，1H)，3.18-3.16(m，2H)，2.92-2.89(d，J＝9Hz，2H)，1.80(m，2H)，1.59-1.57(m，2H)，1.47(m，2H)，1.23(s，22H)，0.87-0.83(t，J＝6.0Hz，3H).According to the method of Example 43, using 580 mg (0.86 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₁₂ CH₃ as raw material, 430 mg of the title compound was obtained as a free Color solid, yield 88%.Mp 139-140°C, ESI-MS (m/z): 534.4[M+H]⁺ .IR (KBr): 3336.49, 3171.37, 2932.91, 1658.63, 1504.92, 1218.19.¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm =9.34(s, 1H), 9.20-9.18(d, J=6Hz, 1H), 8.38(s, 3H), 8.01-7.97(t, J=6Hz J=6Hz, 1H), 7.80(m, 1H) , 7.53(m, 1H), 7.07-7.05(d, J=6Hz, 2H), 6.70-6.67(d, J=9Hz, 2H), 4.42-4.36(Q, J=6Hz 1H), 4.04-3.97( m, 2H), 3.94-3.88(m, 1H), 3.18-3.16(m, 2H), 2.92-2.89(d, J=9Hz, 2H), 1.80(m, 2H), 1.59-1.57(m, 2H ), 1.47(m, 2H), 1.23(s, 22H), 0.87-0.83(t, J=6.0Hz, 3H).

实施例53制备Arg-Tyr-OCH₂(CH₂)₁₄CH₃ (5k)Example 53 Preparation of Arg-Tyr-OCH₂ (CH₂ )₁₄ CH₃ (5k)

按照实施例43的方法，以800mg(1.13mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₁₄CH₃为原料，得标题化合物520mg，为无色固体，产率77％.Mp 143-144℃，

ESI-MS(m/z)：562.5[M+H]⁺.IR(KBr)：3184.48，2929.63.1721.03.1661.55，1513.36，1357.11，1227.39，825.79，570.18.According to the method of Example 43, using 800 mg (1.13 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₁₄ CH₃ as raw material, 520 mg of the title compound was obtained as a free Color solid, yield 77%. Mp 143-144°C,

ESI-MS(m/z): 562.5[M+H]⁺ .IR(KBr): 3184.48, 2929.63.1721.03.1661.55, 1513.36, 1357.11, 1227.39, 825.79, 570.18.

¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.22-9.19(d，J＝9Hz，1H)，8.48-8.47(d，J＝3Hz，1H)，8.43-8.42(m，1H)，8.39(s，1H)，8.00(m，1H)，7.55(s，1H)，7.22(m，1H)，7.07-7.04(d，J＝9Hz，2H)，6.70-6.67(d，J＝9Hz，2H)，4.42-4.35(m，1H)，4.07-3.96(m，2H)，3.94-3.91(m，1H)，3.17(s，2H)，2.91-2.89(d，J＝6Hz，2H)，1.80-1.78(m，2H)，1.60-1.57(m，2H)，1.46(m，2H)，1.22(s，26H)，0.86-0.82(t，J＝6.0Hz，3H).¹ H-NMR (300MHz, DMSO-d₆ ): δ/ppm=9.22-9.19(d, J=9Hz, 1H), 8.48-8.47(d, J=3Hz, 1H), 8.43-8.42(m, 1H ), 8.39(s, 1H), 8.00(m, 1H), 7.55(s, 1H), 7.22(m, 1H), 7.07-7.04(d, J=9Hz, 2H), 6.70-6.67(d, J =9Hz, 2H), 4.42-4.35(m, 1H), 4.07-3.96(m, 2H), 3.94-3.91(m, 1H), 3.17(s, 2H), 2.91-2.89(d, J=6Hz, 2H), 1.80-1.78(m, 2H), 1.60-1.57(m, 2H), 1.46(m, 2H), 1.22(s, 26H), 0.86-0.82(t, J=6.0Hz, 3H).

实施例54制备Arg-Tyr-OCH₂(CH₂)₁₆CH₃ (5l)Example 54 Preparation of Arg-Tyr-OCH₂ (CH₂ )₁₆ CH₃ (5l)

按照实施例43的方法，以850mg(1.17mmol)N^α-Boc-N^G-Arg(NO₂)-Tyr-OC-H₂(CH₂)₁₆CH₃为原料，得标题化合物640mg，为无色固体，产率87％.Mp 146-147℃，

ESI-MS(m/z)：590.6[M+H]⁺.IR(KBr)：3186.89，2928.04，1739.68，1661.65，1513.79，1356.93，1225.64，1111.31，825.40，565.51.¹H-NMR(300MHz，DMSO-d₆)：δ/ppm＝9.38(s，1H)，9.19-9.17(d，J＝6Hz，2H)，8.47-8.45(d，J＝6Hz，1H)，8.42-8.41(m，1H)，8.00(m，1H)，7.63-7.57(t，J＝9Hz，1H)，7.46-7.40(t，J＝9Hz，1H)，7.30-7.23(m，1H)，7.07-7.04(d，J＝9Hz，2H)，6.70-6.68(d，J＝6Hz，2H)，4.42-4.37(m，1H)，4.07-4.00(m，2H)，3.98-3.93(m，1H)，3.26(m，2H)，2.92-2.89(d，J＝9Hz，2H)，1.80-1.75(m，2H)，1.56-1.55(m，2H)，1.49-1.46(m，2H)，1.23(s，30H)，0.86-0.82(t，J＝6.0Hz，3H).According to the method of Example 43, using 850 mg (1.17 mmol) N^α -Boc-N^G -Arg(NO₂ )-Tyr-OC-H₂ (CH₂ )₁₆ CH₃ as raw material, 640 mg of the title compound was obtained as a free Color solid, yield 87%. Mp 146-147°C,

ESI-MS (m/z): 590.6[M+H]⁺ .IR (KBr): 3186.89, 2928.04, 1739.68, 1661.65, 1513.79, 1356.93, 1225.64, 1111.31, 825.40, 565.51.¹ H-NMR (300MHz, DMSO -d₆ ): δ/ppm=9.38(s, 1H), 9.19-9.17(d, J=6Hz, 2H), 8.47-8.45(d, J=6Hz, 1H), 8.42-8.41(m, 1H) , 8.00(m, 1H), 7.63-7.57(t, J=9Hz, 1H), 7.46-7.40(t, J=9Hz, 1H), 7.30-7.23(m, 1H), 7.07-7.04(d, J =9Hz, 2H), 6.70-6.68(d, J=6Hz, 2H), 4.42-4.37(m, 1H), 4.07-4.00(m, 2H), 3.98-3.93(m, 1H), 3.26(m, 2H), 2.92-2.89(d, J=9Hz, 2H), 1.80-1.75(m, 2H), 1.56-1.55(m, 2H), 1.49-1.46(m, 2H), 1.23(s, 30H), 0.86-0.82(t, J=6.0Hz, 3H).

试验例1本发明的化合物5a-l的镇痛活性实验Test example 1 The analgesic activity experiment ofcompound 5a-1 of the present invention

1)受试化合物：本发明的化合物5a-l1) Test compound: compound 5a-1 of the present invention

阳性对照品：阿司匹林；Positive control substance: aspirin;

2)实验动物：ICR小鼠，雄性，体重20±2g；每12只小鼠一组，空白及阳性对照各一组。2) Experimental animals: ICR mice, male, weighing 20±2g; every group of 12 mice, each group of blank and positive control.

3)剂量设置：阿司匹林165μmol/kg，Arg-Tyr 100μmol/kg，5a-l 10μmol/kg，均单次灌胃。3) Dosage setting: Aspirin 165 μmol/kg, Arg-Tyr 100 μmol/kg, 5a-l 10 μmol/kg, all administered by single gavage.

药物配制：生理盐水(NS)溶解。Drug preparation: dissolved in normal saline (NS).

4)给药方案：0.2ml溶液/鼠。4) Dosing regimen: 0.2ml solution/mouse.

5)动物模型5) Animal model

ICR雄性小鼠使用前静息1天，操作间保持室内温度22℃，每组小鼠12只。基础痛阈的测定是在开始时，先测三次，每次间隔5分钟，取其平均值为基础痛阈。每次试验设生理盐水组为平行对照。ICR male mice rested for 1 day before use, and the room temperature was kept at 22°C in the operating room, with 12 mice in each group. The basic pain threshold was measured three times at the beginning, with an interval of 5 minutes each time, and the average value was taken as the basic pain threshold. In each experiment, the normal saline group was set as a parallel control.

6)痛阈提高率的测定6) Determination of pain threshold improvement rate

灌胃后每30min测定一次痛阈值，共测定6组180min。痛阈改变以下列公式表示：痛阈提高％＝[(给药后痛阈-基础痛阈)/基础痛阈]×100％。The pain threshold was measured every 30 minutes after intragastric administration, and a total of 6 groups were measured for 180 minutes. The change of pain threshold is expressed by the following formula: Pain threshold increase %=[(pain threshold after administration-basic pain threshold)/basic pain threshold]×100%.

7)统计方法7) Statistical methods

本实验数据统计均采用t检验和方差分析，以X±SD表示。The statistics of the experimental data were all used t test and analysis of variance, expressed as X±SD.

8)实验结果8) Experimental results

受试化合物热辐射甩尾实验痛阈提高结果如表1所示。在10μmol/kg剂量下受试化合物经灌胃给药后对疼痛有显著的抑制作用。Table 1 shows the pain threshold improvement results of the tested compounds in the thermal radiation tail flick test. At a dose of 10 μmol/kg, the test compound has a significant inhibitory effect on pain after intragastric administration.

表1化合物5a-l的镇痛作用The analgesic effect of table 1compound 5a-l

Aspirin剂量：165μmol/kg；Arg-Tyr剂量：100μmol/kg；5a-l剂量：10μmol/kg.a)与NS组比较P＜0.01；b)与NS组比较P＜0.05；c)与NS组比较P＜0.01，与Arg-Tyr组比较P＜0.01；d)与NS组比较P＜0.01，与Arg-Tyr组比较P＜0.05；e)与NS组比较P＜0.01，与Arg-Tyr组比较P＜0.01，与Aspirin比较P＜0.01；f)与NS组比较P＜0.01，与Arg-Tyr组比较P＜0.01，与Aspirin比较P＜0.05；g)与NS组比较P＜0.01，与Arg-Tyr组比较P＜0.05，与Aspirin比较P＜0.01；h)与NS组比较P＜0.01，与Aspirin比较P＜0.01；i)与NS组比较P＜0.01，与Aspirin比较P＜0.05；j)与NS组比较P＜0.05；与Arg-Tyr组比较P＜0.05Aspirin dose: 165 μmol/kg; Arg-Tyr dose: 100 μmol/kg; 5a-l dose: 10 μmol/kg. a) Compared with NS group, P<0.01; b) Compared with NS group, P<0.05; c) Compared with NS group Compared P<0.01, compared with Arg-Tyr group, P<0.01; d) compared with NS group, P<0.01, compared with Arg-Tyr group, P<0.05; e) compared with NS group, P<0.01, compared with Arg-Tyr group Compared P<0.01, compared with Aspirin, P<0.01; f) compared with NS group, P<0.01, compared with Arg-Tyr group, P<0.01, compared with Aspirin, P<0.05; g) compared with NS group, P<0.01, compared with P<0.05 compared with Arg-Tyr group, P<0.01 compared with Aspirin; h) P<0.01 compared with NS group, P<0.01 compared with Aspirin; i) P<0.01 compared with NS group, P<0.05 compared with Aspirin; j) Compared with NS group, P<0.05; compared with Arg-Tyr group, P<0.05

试验例2本发明化合物5c，5l的镇痛活性剂量和效应的关系Test Example 2 Compound 5c of the present invention, the relationship between the analgesic activity dose and effect of 5l

试验方法同试验例1，受试化合物为5c，5l；将受试化合物分别按10μmol/kg，1μmol/kg和0.1μmol/kg剂量，均采用灌胃单次给药。试验结果见表2。结果表明，本发明化合物5c，5l的镇痛活性呈现剂量依赖关系。The test method was the same as that in Test Example 1, the test compounds were 5c and 5l; the test compounds were dosed at 10 μmol/kg, 1 μmol/kg and 0.1 μmol/kg, respectively, and were administered by intragastric administration in a single dose. The test results are shown in Table 2. The results show that the analgesic activity of compounds 5c and 5l of the present invention exhibits a dose-dependent relationship.

表2不同剂量5c，5l的镇痛活性Table 2 Different doses of 5c, the analgesic activity of 5l

n＝12；受试化合物5c，5l的剂量分别为H＝10μmol/kg，M＝1μmol/kg或L＝0.1μmol/kg.a)与NS组比较p＜0.01；b)与NS组比较p＜0.05；c)与NS组比较p＜0.01，与M组比较p＜0.01；d)与NS组比较p＜0.01，与1μmol/kg组比较p＜0.05.n=12; the doses of test compounds 5c and 5l were H=10 μmol/kg, M=1 μmol/kg or L=0.1 μmol/kg. a) compared with NS group p<0.01; b) compared with NS group p <0.05; c) p<0.01 compared with NS group, p<0.01 compared with M group; d) p<0.01 compared with NS group, p<0.05 compared with 1μmol/kg group.

实验例3本发明化合物5a-l的抗炎活性评价Experimental example 3 Anti-inflammatory activity evaluation ofcompound 5a-1 of the present invention

化合物5a-l或阿司匹林用生理盐水配成溶液，雄性ICR小鼠(20±2g)随机分为空白对照组、阿司匹林组、Arg-Tyr及5a-l组，每组12只小鼠。分别灌胃给予化合物5a-l(剂量为10μmol/kg)或Arg-Tyr(剂量为100μmol/kg)或阿司匹林(剂量为165μmol/kg)溶液0.2ml/只，给药30分钟后，往小白鼠的左耳外廓涂0.03ml二甲苯。2小时后将小白鼠颈椎脱臼处死、两耳用直径7mm的打孔器取圆形耳片、称重、把两圆耳片的重量差作为肿胀度。结果列入表3。实验结果表明，本发明的化合物5a-l具有明确的抗炎活性Compound 5a-l or aspirin was made into a solution with physiological saline, and male ICR mice (20±2g) were randomly divided into blank control group, aspirin group, Arg-Tyr and 5a-l groups, with 12 mice in each group. Intragastric administration ofcompound 5a-l (dose of 10 μmol/kg) or Arg-Tyr (dose of 100 μmol/kg) or aspirin (dose of 165 μmol/kg) solution 0.2ml/only, 30 minutes after administration, injected Apply 0.03ml of xylene to the outside of the left ear. After 2 hours, the mice were sacrificed by cervical dislocation, and round ear pieces were taken from both ears with a puncher with a diameter of 7 mm, weighed, and the weight difference between the two round ear pieces was used as the degree of swelling. The results are listed in Table 3. Experimental results show thatcompound 5a-1 of the present invention has definite anti-inflammatory activity

表3化合物5a-l对小鼠耳肿胀的抑制作用The inhibitory effect of table 3compound 5a-l on mouse ear swelling

阿司匹林剂量为165μmol/kg，Arg-Tyr剂量为100μmol/kg；5m-x剂量10μmol/kg；Aspirin dose is 165 μmol/kg, Arg-Tyr dose is 100 μmol/kg; 5m-x dose is 10 μmol/kg;

n＝12，a)与NS组比较P＜0.01；b)与NS组比较P＜0.01，与Aspirin组比较p＜0.05；c)与NS组比较P＜0.01，与Arg-Tyr组比较p＜0.05；d)与NS组比较P＜0.01，与Aspirin组比较p＜0.05，与Arg-Tyr组比较p＜0.05n=12, a) P<0.01 compared with NS group; b) P<0.01 compared with NS group, p<0.05 compared with Aspirin group; c) P<0.01 compared with NS group, p<0.01 compared with Arg-Tyr group 0.05; d) P<0.01 compared with NS group, p<0.05 compared with Aspirin group, p<0.05 compared with Arg-Tyr group

实验例4不同剂量5c，5l的抗炎活性评价Experimental example 4 different doses 5c, anti-inflammatory activity evaluation of 5l

按照实验例3的方法，5c，5l均按10μmol/kg、1μmol/kg、0.1μmol/kg三种剂量给雄性ICR小鼠灌胃，两圆耳片的重量差列入表4。结果表明本发明的化合物5c，5l的抗炎活性具有剂量依赖性。According to the method of Experimental Example 3, 5c and 5l were administered to male ICR mice in three doses of 10 μmol/kg, 1 μmol/kg, and 0.1 μmol/kg, and the weight difference between the two round ears is listed in Table 4. The results show that the anti-inflammatory activity of compounds 5c and 5l of the present invention is dose-dependent.

表4不同剂量5c，5l对小鼠耳肿胀的抑制作用The different doses of table 4 5c, the inhibitory effect of 5l on mouse ear swelling

a)与NS组比较p＜0.01；b)与NS组比较p＜0.01，与1μmol/kg组比较p＜0.01.a) p<0.01 compared with NS group; b) p<0.01 compared with NS group, p<0.01 compared with 1 μmol/kg group.

以上所述的实施例仅仅是对本发明的优选实施方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案作出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。The above-mentioned embodiments are only descriptions of preferred implementations of the present invention, and are not intended to limit the scope of the present invention. Variations and improvements should fall within the scope of protection defined by the claims of the present invention.

Claims

1. the compound that general formula is I:

Arg-Tyr-XCH₂(CH₂)_nCH₃

I

In formula, X is NH or O;

In formula, n is 6,8,10,12,14 or 16.

2. a method of preparing compound claimed in claim 1, is characterized in that comprising the following steps:

(1) under the vitriol oil, concentrated nitric acid condition, Arg nitration obtains N^g-Arg (NO₂);

(2) under alkalescent water dioxane condition, N^g-Arg (NO₂) and (Boc)₂o reaction makes N^α-Boc-N^g-Arg (NO₂), L-Tyr with (Boc)₂o reaction makes Boc-Tyr;

(3) under DCC and HOBt existence, Boc-Tyr is condensed into Boc-Tyr-XCH with aliphatic amide or fatty alcohol in anhydrous THF₂(CH₂)_ncH₃;

(4) in the ethyl acetate of containing hydrogen chloride, Boc-Tyr-XCH₂(CH₂)_ncH₃de-Boc protection, obtains Tyr-XCH₂-(CH₂)_ncH₃;

(5) under DCC and HOBt existence, N^α-Boc-N^g-Arg (NO₂) in anhydrous THF with Boc-Tyr-X-CH₂(CH₂)_ncH₃condensation, prepares N^α-Boc-N^g-Arg (NO₂)-Tyr-XCH₂(CH₂)_ncH₃;

(6) H₂n under/Pd condition^α-Boc-N^g-Arg (NO₂)-Tyr-XCH₂(CH₂)_ncH₃de--NO₂protection, obtains Boc-Arg-Tyr-XCH₂(CH₂)_ncH₃;

(7) in the ethyl acetate of containing hydrogen chloride, Boc-Arg-Tyr-XCH₂(CH₂)_ncH₃de-Boc protection, obtains Arg-Tyr-XCH₂(CH₂)_ncH₃.

3. method according to claim 2, is characterized in that: the aliphatic amide described in described step (3) is positive eight amine, positive ten amine, n-dodecylamine, positive tetradecy lamine, hexadecylamine or octadecyl amine; Described fatty alcohol is positive eight alcohol, positive ten alcohol, n-dodecanol, tetradecanol, positive hexadecanol or positive stearyl alcohol.

4. method according to claim 2, is characterized in that: described Arg and Tyr are L configuration amino acid.

5. the purposes of compound claimed in claim 1 in preparation analgesia and anti-inflammatory drug.