TW201534298A

TW201534298A - FPR1 antagonist derivatives and use thereof (2)

Info

Publication number: TW201534298A
Application number: TW104105442A
Authority: TW
Inventors: Tsong-Long Hwang; Pei-Wen Hsieh; yin-ting Huang; Chih-Hao Hung
Original assignee: Univ Chang Gung
Priority date: 2013-10-09
Filing date: 2013-10-09
Publication date: 2015-09-16

Abstract

Binding of fomyl peptide receptor 1 (FPR1) by N-formyl peptides can induce neutrophil activation and may represent a new therapeutic target in either sterile or septic inflammation. In present application, a series of new dipeptide derivatives as shown general formula (I) is provided. Of these, each of R1 and R4 is one selected from a group consisting of hydrogen atom, hydroxyl group, C1-C4 alkyl, C1-C4 alkoxyl and glycin-nitrile amide group; each of R2 and R3 is one selected from a group consisting of hydrogen atom, hydroxyl group, pyridinyl group, phenyl group, pyridinyl group, C1-C4 alkoxyl substitute on the ester group, and a hydroxyl group, halogen group, C1-C4 alkoxyl group, C1-C4 alkyl group substitute on the aromatic ring of benzoyl group. Additionally, the configurations of two amino acids are S and R configurations, respectively.

Description

FPR1拮抗劑的衍生物及其用途(二) Derivatives of FPR1 antagonists and their uses (2)

本申請相關於甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)拮抗劑之雙胜肽類衍生物N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters)，及其用於治療涉及FPR1活性之病症。 The present application relates to a double peptide derivative N- ( N -aralkyl-L-chromocarbyl)-D- of a Formyl Peptide Receptor 1 (FPR1) antagonist. N- ( N- aroyl-L-tryptophanyl-D-phenylalanine methyl esters), and their use in the treatment of conditions involving FPR1 activity.

甲醯化胜肽受體(Formyl peptide receptor,FPR)屬於G蛋白偶聯受體(GPCR)的一種，在人體中FPR受體家族可分成三類；分別為FPR1、FPR2、FPR3。而FPR2與FPR3納入類甲醯化胜肽受體，FPR2又稱為類甲醯化胜肽受體1(FPR-like receptor 1,FPRL-1)，FPR3則稱為類甲醯化胜肽受體2(FPRL-2)。FPR1多存在於單核細胞、多形核白血球(PMN)以及未成熟樹突細胞；而FPR2則存在於肝細胞、肺臟細胞、脾臟細胞、T淋巴球、多形核白血球及單核細胞，其中嗜中性白血球只表現FPR1和FPR2。N-甲醯甲硫氨醯亮氨醯苯丙氨酸(N-formyl-L-methionyl-L-leucyl-L-phenylalanine,fMLP或fMIF)屬於N-甲醯胜肽，為一常用於結合在FPR的化學趨化因子進而啟動細胞活化反應並釋放毒殺物質或蛋白酶。fMLF對於FPR三種受體的親和性不同，其中對於FPR1具有較高的親和性(FPR1的EC₅₀為0.1-1nM；FPR2為1M)。當FPR1與配體結合時，使得G蛋白活化，並且依細胞不同可活化下游不同的訊息傳遞路徑，比如鈣離子流、磷脂酵素毒素C、磷脂醯肌醇3-激酶(PI3K)、有絲***活化蛋白質激酶(mitogen-activated protein kinases,MAPKs)及蛋白酪胺酸激酶，使得嗜中性白血球活化進行轉移、呼吸暴破及去顆粒化等作用。因此有文獻指出，FPR1拮抗劑可作為治療嗜中性白血球所誘發的各種發炎疾病；比如氣喘(asthma)、類風濕性關節炎(rheumatoid arthritis)、乾癬(psoriasis)等。此外，其他文獻亦指FPR1不當的調控會造成牙周炎(periodontitis)、心血管疾病、動脈粥狀硬化、胃癌等疾病。而近年來的研究指出，FPR1能與內生性膜聯蛋白(annexin AI)作用，而活化膠質母細胞瘤癌細胞中表皮生長因子(EGFR)的表現，進而影響該腫瘤細胞的生長與轉移。因此FPR1也可做為膠質母細胞瘤(glioblastoma)的治療標的。 Formyl peptide receptor (FPR) belongs to G protein-coupled receptor (GPCR). In the human body, FPR receptor family can be divided into three categories; FPR1, FPR2 and FPR3. FPR2 and FPR3 are involved in the hyperthyroidism receptor, FPR2 is also known as FPR-like receptor 1 (FPRL-1), and FPR3 is called hyperthyroidism. Body 2 (FPRL-2). FPR1 is mostly present in monocytes, polymorphonuclear white blood cells (PMN) and immature dendritic cells; while FPR2 is present in hepatocytes, lung cells, spleen cells, T lymphocytes, polymorphonuclear leukocytes and monocytes, of which Neutrophilic white blood cells only exhibit FPR1 and FPR2. N - acyl carboxylic acyl methionyl leucine phenylalanine acyl (N -formyl-L-methionyl- L-leucyl-L-phenylalanine, fMLP or FMIF) belonging to N - carboxylic acyl peptide, is a commonly used binding The chemical chemokine of FPR in turn initiates a cell activation reaction and releases a toxic substance or protease. fMLF affinity for the FPR different three receptors, which have a high affinity for FPR1 (FPR1 the EC ₅₀ is 0.1-1nM; FPR2 of 1M). When FPR1 binds to a ligand, it activates the G protein and activates different downstream signaling pathways depending on the cell, such as calcium ion flux, phospholipase toxin C, phospholipid creatinine 3-kinase (PI3K), mitotically activated protein. Mitogen-activated protein kinases (MAPKs) and protein tyrosine kinases cause neutrophil activation, metastasis, and granulation. Therefore, it has been pointed out in the literature that FPR1 antagonists can be used as various inflammatory diseases induced by neutrophils; for example, asthma, rheumatoid arthritis, psoriasis, and the like. In addition, other literature also refers to the improper regulation of FPR1 can cause periodontitis (periodontitis), cardiovascular disease, atherosclerosis, gastric cancer and other diseases. In recent years, studies have shown that FPR1 can interact with endogenous annexin (AI) and activate epidermal growth factor (EGFR) in glioblastoma cells, which in turn affects the growth and metastasis of tumor cells. Therefore, FPR1 can also be used as a therapeutic target for glioblastoma.

在2010年，Movitz等人的研究指出在多胜肽C端具有色胺酸(tryptophan)與***酸(phenylalanine)所形成的雙肽能夠協助此多胜肽選擇性地與FPR1受體結合；然而，此雙肽並無法抑制N-甲醯甲硫氨醯亮氨醯苯丙氨酸(fMLF)所誘導的嗜中性白血球細胞的呼吸***反應及其相關的超氧自由基的生成。 In 2010, Movitz et al. showed that dipeptides with tryptophan and phenylalanine at the C-terminus of the multi-peptide help the multi-peptide to selectively bind to the FPR1 receptor; This dipeptide does not inhibit the respiratory burst reaction of neutrophil cells induced by N -methylpyrazine and leucine phenylalanine (fMLF) and the formation of related superoxide radicals.

另外，經由Google專利與Orbit專利資料庫搜尋得知，具有編號EP2490021 A1與WO2012112048 A1等的專利文件載明一系列具有化學式如H-X₁-X₂-OH之雙胜肽類衍生物，其可作用於模式識別受體與G蛋白偶聯受體訊息傳遞路徑。其中兩胺基酸需皆為L(或S)構型。 In addition, through the search of the Google patent and the Orbit patent database, the patent documents having the numbers EP2490021 A1 and WO2012112048 A1 and the like contain a series of double-peptide derivatives having a chemical formula such as HX ₁ -X ₂ -OH, which can act. The pattern recognition receptor and G protein-coupled receptor signaling pathway. The two amino acids need to be in the L (or S ) configuration.

具有編號US20130109866 A1與WO2013062947 A1的專利文件載明一系列N端具有尿素基團的胺基酸衍生物，其可調節FPRL-1受體，此FPRL-1受體為FPR2受體的別名。 The patent documents with the numbers US20130109866 A1 and WO2013062947 A1 contain a series of N-terminal amino acid-containing amino acid derivatives which modulate the FPRL-1 receptor, which is an alias for the FPR2 receptor.

具有編號WO2012074785 A1、WO2013070600 A1、US8440684 B2、WO2013009543 A1、US20120238628 A1、US20110319454 A1與WO2012109544 A1的專利文件皆載明調節FPRL-1受體化合物的技術方案。 The patent documents for the regulation of FPRL-1 receptor compounds are all described in the patent documents with the numbers WO2012074785 A1, WO2013070600 A1, US8440684 B2, WO2013009543 A1, US20120238628 A1, US20110319454 A1 and WO2012109544 A1.

由上述可知，FPR1拮抗劑可以調控發炎與腫瘤等疾病，且目前臨床上並無FPR1拮抗劑，故發展FPR1拮抗劑是目前極為重要的。 From the above, it can be known that FPR1 antagonists can regulate diseases such as inflammation and tumors, and currently there is no FPR1 antagonist in clinical practice, so development of FPR1 antagonists is extremely important at present.

因此，本發明主要提供一種如式一所示的雙胜肽類衍生物，其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；其中R1、R4分別選自氫基(hydrogen atom)、羥基(hydroxyl group)、C₁-C₄烷基取代之羥基、C₁-C₄烷氧基(alkoxyl)、羧酸基團(carboxylic acids group)，醯胺基團含有C₁-C₄烷基(alkyl)氰基之取代、或C₁-C₄烷基或C₁-C₄烷氧基之取代，酯基團含有C₁-C₄烷基(alkyl)之取代或苯環上含有C₁-C₄烷基(alkyl)取代之苯醯基團(benzoyl group)其中之一；R2、R3分別選自氫基、羥基、苯基(phenyl group)、吡啶(pyridinyl)、羧酸基團(carboxylic acids group)，或是含有C₁-C₄烷氧基之酯基團，或是在苯環上含有羥基、鹵素、C₁-C₄烷氧基(alkoxyl)或C₁-C₄烷基(alkyl)取代之苯醯基團(benzoyl group)。其中鹵素係氟、氯、溴、碘等基團。 Therefore, the present invention mainly provides a double peptide derivative as shown in Formula 1, wherein: the chiral centers of the compound of the formula 1 are S configuration and R configuration, respectively; wherein R1 and R4 are respectively Selected from a hydrogen atom, a hydroxyl group, a C ₁ -C ₄ alkyl substituted hydroxyl group, a C ₁ -C ₄ alkoxyl group, a carboxylic acid group, a guanamine The group contains a C ₁ -C ₄ alkyl cyano group, or a C ₁ -C ₄ alkyl group or a C ₁ -C ₄ alkoxy group, the ester group containing a C ₁ -C ₄ alkyl group ( a substitution of alkyl) or a phenyl ring containing a C ₁ -C ₄ alkyl substituted benzoyl group; R 2 and R 3 are each selected from a hydrogen group, a hydroxyl group, or a phenyl group. Pyridinyl, carboxylic acid acid group, or an ester group containing a C ₁ -C ₄ alkoxy group, or a hydroxyl group, a halogen, a C ₁ -C ₄ alkoxy group on the benzene ring Alkoxyl or a C ₁ -C ₄ alkyl substituted benzoyl group. Among them, halogen is a group such as fluorine, chlorine, bromine or iodine.

本發明亦揭露雙胜肽類衍生物對於抑制fMLF所誘導的嗜中性白血球彈性蛋白酶與超氧自由基的生成具有極佳的抑制作用，且具競爭性抑制FPR1下游的訊息傳遞路徑，包含鈣離子、MAPKs以及蛋白質激酶B(protein Kinase B,Akt)。實驗結果指出上述雙胜肽類衍生物具備做為FPR1拮抗劑的潛力。因此本發明所載之雙胜肽類衍生物可進一步開發作為抗發炎疾病藥物。 The present invention also discloses that the double peptide derivative has an excellent inhibitory effect on inhibiting the formation of neutrophil elastase and superoxide radical induced by fMLF, and competitively inhibits the signal transduction pathway downstream of FPR1, including calcium. Ions, MAPKs, and protein kinase B (Akt). The experimental results indicate that the above-mentioned double peptide derivatives have the potential to be antagonists of FPR1. Therefore, the double peptide derivative contained in the present invention can be further developed as an anti-inflammatory disease drug.

R₁~R₄‧‧‧雙胜肽類衍生物中的取代基 Substituents in R ₁ ~R ₄ ‧‧‧bis peptide derivatives

化合物3-24‧‧‧雙胜肽類衍生物 Compound 3-24 ‧‧‧Double peptide derivatives

Sivelestat,EFB-1,Cyclosporin(CycH),WRW4‧‧‧正向控制組 Sivelestat, EFB-1, Cyclosporin (CycH), WRW4‧‧‧ Forward Control Group

Cs H‧‧‧Cyclosporin H Cs H‧‧‧Cyclosporin H

DMSO‧‧‧二甲基亞碸 DMSO‧‧ dimethyl hydrazine

HCH6-1‧‧‧雙胜肽類衍生物之某一實施例 An embodiment of HCH6-1‧‧‧ double peptide derivatives

FNLFNYK‧‧‧N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein FNLFNYK‧‧‧N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein

MAPKs(ERK、p38和JNK)及Akt‧‧‧訊息傳遞路徑 MAPKs (ERK, p38 and JNK) and Akt‧‧‧ message delivery paths

圖1(a)~圖1(b)繪示HCH6-1對於人類嗜中性白血球釋放乳酸脫氫酶影響；圖2(a)~圖2(f)繪示HCH6-1選擇性抑制fMLF引發人類嗜中性白血球CD11b表現；圖3(a)~圖3(b)繪示HCH6-1抑制FNLFNYK結合在人類嗜中性白血球膜上的FPR1的效應；圖4(a)~圖4(d)繪示HCH6-1具有顯著的抑制MAPKs(ERK、p38和JNK)及Akt磷酸化。 Figure 1 (a) ~ Figure 1 (b) shows the effect of HCH6-1 on the release of lactate dehydrogenase from human neutrophils; Figure 2 (a) ~ Figure 2 (f) shows that HCH6-1 selectively inhibits fMLF initiation Human neutrophil CD11b expression; Figure 3 (a) ~ Figure 3 (b) shows that HCH6-1 inhibits FNLFNYK binding in humans Effect of FPR1 on neutrophils; Figures 4(a) to 4(d) show that HCH6-1 has significant inhibition of MAPKs (ERK, p38 and JNK) and Akt phosphorylation.

本發明主要提供一種如式一所示的雙胜肽類衍生物，其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型； The present invention mainly provides a double peptide derivative as shown in Formula 1, wherein: the chiral centers of the compound of the formula 1 are S configuration and R configuration, respectively;

其中該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；其中R1、R4分別選自氫基(hydrogen atom)、羥基(hydroxyl group)、C₁-C₄烷基取代之羥基、C₁-C₄烷氧基(alkoxyl)、羧酸基團(carboxylic acids group)，醯胺基團含有C₁-C₄烷基(alkyl)氰基之取代、或C₁-C₄烷基或C₁-C₄烷氧基之取代，酯基團含有C₁-C₄烷基(alkyl)之取代或苯環上含有C₁-C₄烷基(alkyl)取代之苯醯基團(benzoyl group)其中之一；R2、R3分別選自氫基、羥基、苯基(phenyl group)、吡啶(pyridinyl)、羧酸基團(carboxylic acids group)，或是含有C₁-C₄烷氧基之酯基團，或是在苯環上含有羥基、鹵素、C₁-C₄烷氧基(alkoxyl)或C₁-C₄烷基(alkyl)取代之苯醯基團(benzoyl group)。其中鹵素係氟、氯、溴、碘等基團。 Wherein the chiral centers of the compound of the formula are S configuration and R configuration, respectively; wherein R1 and R4 are respectively selected from a hydrogen atom, a hydroxyl group, and a C ₁ -C ₄ alkane. a substituted hydroxy group, a C ₁ -C ₄ alkoxyl group, a carboxylic acid group, a guanamine group containing a C ₁ -C ₄ alkyl cyano group, or a C _{1 group} a substitution of a -C ₄ alkyl group or a C ₁ -C ₄ alkoxy group, the ester group containing a C ₁ -C ₄ alkyl group or a phenyl ring containing a C ₁ -C ₄ alkyl group substituted One of a benzoyl group; R2 and R3 are each selected from a hydrogen group, a hydroxyl group, a phenyl group, a pyridinyl group, a carboxylic acid group, or a C ₁ group _; An ester group of a -C ₄ alkoxy group, or a benzoquinone group having a hydroxyl group, a halogen, a C ₁ -C ₄ alkoxyl group or a C ₁ -C ₄ alkyl group substituted on the benzene ring (benzoyl group). Among them, halogen is a group such as fluorine, chlorine, bromine or iodine.

本案之雙胜肽類衍生物N-(N-苄基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-Benzoyl-L-tryptophanyl)-D-phenylanlanine methyl ester，HCH6-1)是以下列合成方法：流程(scheme)一、流程二、流程三或流程四而得出。流程一可合成化合物2-13，流程二可合成化合物16-24，流程三可合成化合物25-27以及流程四可合成化合物14-15，上述化合物的每一化合物或可進一步區分為主要(major)或次要(minor)產物，於化合物編碼註記主要產物為a與次要產物為b，R₁~R₄為取代基，在藥學上可接受的鹽類溶劑合物或其組合亦可加入雙胜肽類衍生物之中。 Double Chance case the peptide derivative N - (N - benzyl-amino acyl -L- color) -D- phenylalanine methyl ester (N - (N -Benzoyl-L -tryptophanyl) -D-phenylanlanine methyl ester , HCH6-1) is derived from the following synthesis methods: scheme 1, process 2, process 3 or process 4. Scheme 1 can synthesize compound 2-13 , Scheme 2 can synthesize compound 16-24 , Scheme 3 can synthesize compound 25-27, and Scheme 4 can synthesize compound 14-15 . Each compound of the above compound can be further divided into major (major) Or a minor product, wherein the main product is a and the minor product is b, and R ₁ to R ₄ are a substituent, and a pharmaceutically acceptable salt solvate or a combination thereof may also be added. Among the double peptide derivatives.

流程一是製備N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters)以及其相關衍生物的方法，可合成化合物2-13。該流程一的試劑與反應條件如下：(a)2.0N氫氧化鈉(NaOH)/芳香醯氯(aroyl chlorides)/20小時；(b)D-苯丙氨酸甲酯鹽酸鹽(D-phenylalanine methyl ester)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)/N,N-二異丙基乙胺(DIEA)/二氯甲烷(DCM)6小時；(c)2.0N氫氧化鈉(NaOH)/苯甲醯氯(benzoyl chloride)20小時；(d)D-苯丙氨醇(D-phenylalaninol)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)/N,N-二異丙基乙胺(DIEA)/二氯甲烷(DCM)6小時。 The first step is to prepare N- ( N -aryl-L-tryptophanyl-D-phenylalanine methyl esters) and the N- ( N- aroyl-L-tryptophanyl)-D-phenylalanine methyl esters thereof. Compounds 2-13 can be synthesized by methods related to derivatives. The reagents and reaction conditions of the first scheme are as follows: (a) 2.0 N sodium hydroxide (NaOH) / aromatic aroyl chlorides / 20 hours; (b) D-phenylalanine methyl ester hydrochloride (D- Phenylalanine methyl ester)/benzotriazole- N,N,N',N' -tetramethyluronium hexafluorophosphate (HBTU) / N,N -diisopropylethylamine (DIEA)/dichloromethane (DCM) 6 hours; (c) 2.0 N sodium hydroxide (NaOH) / benzoyl chloride for 20 hours; (d) D-phenylalaninol / benzotriazole - N,N,N',N '-tetramethyluronium hexafluorophosphate (HBTU) / N,N -diisopropylethylamine (DIEA) / dichloromethane (DCM) for 6 hours.

流程二是製備N-(N-苄基-L-色氨醯基)-對-取代-D-苯丙氨酸甲酯(N-(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine methyl esters)與N-(N-苄基-L-色氨醯基)-3-環己基-D-丙氨酸甲酯(N-(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters)的方法，可合成化合物16-24。該流程二的試劑與反應條件如下：(a)20%三氟乙酸(TFA)(TFA/DCM=1：4)/30mins；(b)甲醇(MeOH)/濃硫酸(c-H₂SO₄)/迴流2小時；(c)N-苯甲醯-左旋色胺酸(N-benzoyl-L-tryptophan)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)/N,N-二異丙基乙胺(DIEA)/二氯甲烷(DCM)6小時。 Second Preparation Process N - (N - benzyl-acyl -L- tryptophyl) - on - substituted -D- phenylalanine methyl ester (N - (N -benzoyl-L -tryptophanyl) - para -substituted-D -phenylalanine methyl esters) with N- ( N -benzyl-L-chromomethyl)-3-cyclohexyl-D-alanine methyl ester ( N -( N -benzoyl-L-tryptophanyl)-3-cyclohexyl The compound 16-24 can be synthesized by the method of -D-alanine methyl esters). The reagents and reaction conditions of the second scheme are as follows: (a) 20% trifluoroacetic acid (TFA) (TFA/DCM = 1:4) / 30 mins; (b) methanol (MeOH) / concentrated sulfuric acid ( c - H ₂ SO ₄ ) / refluxed for 2 hours; (c) N - benzoyl - L-tryptophan (N -benzoyl-L-tryptophan) / benzene Bing triazole - N, N, N ', N' - tetramethyluroniumhexafluorophosphate Fluorophosphate (HBTU) / N,N -diisopropylethylamine (DIEA) / dichloromethane (DCM) for 6 hours.

流程三是製備N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸氨基乙腈(N-(N-benzoyl-L-tryptophanyl)-D-phenylalanine-glycine-nitriles)的方法，可合成化合物25-27。該流程三的試劑與反應條件如下：(a)氨基乙腈(aminoacetonitrile)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(2-(1H-benzotriazol-1-yl-1,1,3,3-tetramethyluronium-hexafluorophosphate，HBTU)/N,N-二異丙基乙胺(N,N-Diisopropylethylamine，DIEA)/二氯甲烷(DCM)6小時；(b)20%三氟乙酸(TFA)與二氯甲烷(DCM)依照TFA/DCM=1：4調配之混合物，30mins；(c)N-苯甲醯-左旋色胺酸(N-benzoyl-L-tryptophan)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)/N,N-二異丙基乙胺(DIEA)/二氯甲烷(DCM)6小時。 Third Preparation Process N - (N - aralkyl acyl -L- tryptophyl) -D- phenylalanine aminoacetonitrile (N - (N -benzoyl-L -tryptophanyl) -D-phenylalanine-glycine-nitriles) The method can synthesize compounds 25-27 . The reagents and reaction conditions of the third scheme are as follows: (a) aminoacetonitrile / benzotriazole - N, N, N', N '-tetramethyl urea hexafluorophosphate (2-(1H-benzotriazol) -1-yl-1,1,3,3-tetramethyluronium- hexafluorophosphate, HBTU) / N, N - diisopropylethylamine (N, N -Diisopropylethylamine, DIEA) / dichloromethane (DCM) 6 hours; ( b) a mixture of 20% trifluoroacetic acid (TFA) and dichloromethane (DCM) according to TFA/DCM=1:4, 30 mins; (c) N -benzidine-L-tryptamine ( N- benzoyl-L) -tryptophan)/benzotriazole- N,N,N',N' -tetramethylurea hexafluorophosphate (HBTU) / N,N -diisopropylethylamine (DIEA) / dichloromethane ( DCM) 6 hours.

流程四是製備N-(N-菸鹼基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-nictotinoyl-L-tryptophanyl)-D-phenylalanine methyl esters)的方法，可合成化合物14-15。該流程四的試劑與反應條件如下：(a)烟醯氯(nicoinoyl chloride)/吡啶(pyridine)/20小時；(b)1.0M氫氧化鋰/四氫呋喃(THF)1 小時；(c)丙烯酸甲酯D-***酸丙烯酸甲酯(D-phenylalanine methyl ester)/苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)/N,N-二異丙基乙胺(DIEA)/二氯甲烷(DCM)6小時。 Process four the preparation of N - (N - nicotine acyl group -L- tryptophyl) -D- phenylalanine methyl ester (N - (N -nictotinoyl-L -tryptophanyl) -D-phenylalanine methyl esters) method , can synthesize compound 14-15 . The reagents and reaction conditions of the scheme 4 are as follows: (a) nicoinoyl chloride / pyridine / 20 hours; (b) 1.0 M lithium hydroxide / tetrahydrofuran (THF) for 1 hour; (c) acrylic acid D-phenylalanine methyl ester/benzotriazole- N,N,N',N '-tetramethyluronium hexafluorophosphate (HBTU) / N,N - diiso Propylethylamine (DIEA) / dichloromethane (DCM) for 6 hours.

本發明中所指賦形劑或稱為『藥學上可接受之載體或賦形劑』、『生物可利用之載體或賦形劑』，係包括溶媒、分散劑、包衣、抗菌或抗真菌劑，保存或延緩吸收劑等任何習知用於製備成劑型之適當化合物。通常此類載體或賦形劑，本身不具備治療疾病之活性，且將本技術所揭示之化合物，搭配藥學上可接受之載體或賦形劑，製備成各劑型，投與動物或人類不致於造成不良反應、過敏或其它不適當反應。因而本技術所揭示之化合物，搭配藥學上可接受之載體或賦形劑，係適用於臨床及人類。『有效劑量』係代表足以改善或防止醫學症狀或生物體狀態之劑量。有效劑量亦說明投與化合物之劑量足供用於診斷之劑量。除非說明書另有敘述，否則『活性化合物』以及『醫藥活性化合物』於此均可替換使用，係指稱一具有製藥學、藥理學或治療以及其他效果之物質。 The excipients referred to in the present invention are referred to as "pharmaceutically acceptable carriers or excipients", "bioavailable carriers or excipients", including solvents, dispersing agents, coatings, antibacterial or antifungal agents. Any suitable compound which is conventionally used in the preparation of a dosage form, such as an absorbent or a delaying absorbent. Usually such carriers or excipients do not themselves have the activity of treating diseases, and the compounds disclosed in the present technology, together with pharmaceutically acceptable carriers or excipients, are prepared into various dosage forms, and are not administered to animals or humans. Cause adverse reactions, allergies or other inappropriate reactions. Thus, the compounds disclosed in the art, in combination with pharmaceutically acceptable carriers or excipients, are suitable for use in clinical and human applications. An "effective dose" is a dose sufficient to ameliorate or prevent a medical condition or a biological condition. The effective dose also means that the dose of the administered compound is sufficient for the diagnosis. Unless otherwise stated in the specification, "active compound" and "pharmaceutically active compound" are used interchangeably herein to refer to a substance having pharmaceutical, pharmacological or therapeutic as well as other effects.

人類嗜中性白血球彈性蛋白酶(elastase)的釋放測定方法：當嗜中性白血球活化時，會由去顆粒化作用的方式釋放出彈性蛋白酶。將含有彈性蛋白酶專一性受質Methoxysuccinyl-Ala-Ala-Pro- Val-p-nitroanilide的嗜中性白血球懸浮液，於37℃與待測化合物作用2分鐘，再加入0.5μg/ml細胞鬆弛素B作用3分鐘來放大反應，最後加入fMLF(30nM)或WKYMVm(1.5nM)反應10分鐘。使用分光光度計於波長405nm下測量吸光值變化，以此評估待測化合物對彈性蛋白酶釋放的影響。測量結果以二甲基亞碸(DMSO)吸光值的改變量作百分比表示。 Method for the release of human neutrophil elastase: When neutrophils are activated, elastase is released by degranulation. Will contain elastase-specific receptor Methoxysuccinyl-Ala-Ala-Pro- A neutrophilic suspension of Val-p-nitroanilide was applied to the test compound for 2 minutes at 37 ° C, and then 0.5 μg / ml of cytochalasin B was added for 3 minutes to amplify the reaction, and finally fMLF (30 nM) or WKYMVm ( 1.5 nM) reaction for 10 minutes. The change in absorbance was measured at a wavelength of 405 nm using a spectrophotometer to evaluate the effect of the test compound on elastase release. The measurement results are expressed as a percentage of the change in the absorbance of dimethyl hydrazine (DMSO).

表一~表六係本案雙胜肽類衍生物、FPR1或FPR2專一性刺激劑所誘導的人類嗜中性白血球彈性蛋白酶(neutrophil elastase)的釋放與超氧自由基(superoxide anion)的生成抑制活性評估結果。 Table 1 to Table 6 are the release of human neutrophil elastase and the superoxide anion production inhibitory activity induced by the double peptide derivative, FPR1 or FPR2 specific stimulator in this case. evaluation result.

表一係N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters及其衍生物，對於由FPR1專一性刺激劑fMLF所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成抑制活性評估結果。 Table 1 is N- ( N- aroyl-L-tryptophanyl)-D-phenylalanine methyl esters and their derivatives, for the release of human neutrophil elastase induced by FPR1 specific stimulator fMLF and superoxide free radicals The resulting inhibitory activity was evaluated.

^a50%抑制所需的濃度IC₅₀表示成平均值±標準誤差(n=3)^bSivelestat & EFB-1為正向控制組^c取自文獻值 ^a 50% inhibition required concentration IC ₅₀ expressed as mean ± standard error (n = 3) ^b Sivelestat & EFB-1 is positive control group ^c taken from the literature value

表二係N-(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine methyl esters與N-(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters對於由FPR1專一性刺激劑fMLF所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成抑制活性評估結果。 Table II Department N - (N -benzoyl-L- tryptophanyl) - para -substituted-D-phenylalanine methyl esters and N - (N -benzoyl-L- tryptophanyl) -3-cyclohexyl-D-alanine methyl esters for the specific FPR1 The results of the evaluation of the release of human neutrophil elastase induced by the sexual stimulator fMLF and the inhibition activity of superoxide radical production.

^a50%抑制所需的濃度IC₅₀表示成平均值±標準誤差(n=3)^bSivelestat & EFB-1為正向控制組 ^a 50% inhibition required concentration IC ₅₀ expressed as mean ± standard error (n = 3) ^b Sivelestat & EFB-1 is the positive control group

表三係N-(N-benzoyl-L-tryptophanyl)-D-phenyl-alanine-glycine-nitriles對於由FPR1專一性刺激劑fMLF所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成抑制活性評估結果。 Table III N- ( N- benzoyl-L-tryptophanyl)-D-phenyl-alanine-glycine-nitriles for the release of human neutrophil elastase induced by the FPR1 specific stimulator fMLF with superoxide radicals The results of the inhibition activity evaluation were generated.

表四係化合物3、6、24a、與24b分別對於由FPR1專一性刺激劑fMLF與FPR2專一性刺激劑WKYMVm所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成抑制活性評估比較結果。 Table 4 Compounds 3 , 6 , 24a , and 24b are evaluated for the release of human neutrophil elastase induced by the FPR1 specific stimulator fMLF and the FPR2 specific stimulator WKYMVm, and the inhibitory activity of superoxide radical production. Comparing results.

^a50%抑制所需的濃度IC₅₀表示成平均值±標準誤差(n=3)^bCyclosporin(CycH)& WRW4為正向控制組 ^a 50% inhibition required concentration IC ₅₀ expressed as mean ± standard error (n = 3) ^b Cyclosporin (CycH) & WRW4 is the positive control group

表五和表六探討雙胜肽衍生物是否具有抗發炎的作用。由於正鐵細胞色素C無法穿透細胞膜，在細胞外與超氧自由基反應，使正鐵細胞色素C還原，且於波長550nm時有吸光反應，以測量吸光值變化的方式來評估待測化合物對超氧自由基釋放的影響；當嗜中性白血球活化時，會經由去顆粒化作用的方式釋放出彈性蛋白酶。表五和表六是利用具有專一性可與彈性白酶作用的受質(Methoxysucciny-Ala-Ala-Pro-Val-p-nitroanilide)，來評估待測化合物影響彈性蛋白酶的釋放。 Tables 5 and 6 discuss whether the double peptide derivative has an anti-inflammatory effect. Since the iron cytochrome C cannot penetrate the cell membrane, it reacts with superoxide radicals outside the cell to reduce the positive iron cytochrome C, and absorbs light at a wavelength of 550 nm. The test compound is evaluated by measuring the change in absorbance. The effect on the release of superoxide radicals; when neutrophils are activated, elastase is released via degranulation. Tables 5 and 6 use Methoxysucciny-Ala-Ala-Pro-Val- p- nitroanilide, which has specificity to interact with elastase, to evaluate the effect of the test compound on the release of elastase.

表五係化合物3(compound HCH6-1)、化合物6(HCH30-2)、化合物19a(HCH108-4)、化合物19b(HCH108-3)、化合物22a(HCH113-4)、化合物22b(HCH113-3)、化合物24a(HCH99-2)、化合物24b(HCH99-1)、化合物27a(HCH90-2-2)和化合物27b(HCH90-2-1)分別對於由FPR1專一性刺激劑fMLF與FPR2專一性刺激劑WKYMVm所誘導的人類嗜中性白血球超氧自由基的生成抑制活性評估比較結果；表六係化合物3(HCH6-1)、化合物6(HCH30-2)、化合物19a(HCH108-4)、化合物19b(HCH108-3)、化合物22a(HCH113-4)、化合物22b(HCH113-3)、化合物24a(HCH99-2)、化合物24b(HCH99-1)、化合物27a(HCH90-2-2)和化合物27b(HCH90-2-1)分別對於由FPR1專一性刺激劑fMLF與FPR2專一性刺激劑WKYMVm所誘導的人類嗜中性白血球彈性蛋白酶的釋放抑制活性評估比較結果。表五和表六之中的IC₅₀表示成平均值±標準誤差(n=4或8)、***p<0.001(相較於控制組)以及Inh%為10μM之下的抑制百分比。 Table 5 is compound HCH6-1, compound 6 (HCH30-2), compound 19a (HCH108-4), compound 19b (HCH108-3), compound 22a (HCH113-4), compound 22b (HCH113-3) ), Compound 24a (HCH99-2), Compound 24b (HCH99-1), Compound 27a (HCH90-2-2), and Compound 27b (HCH90-2-1) are specific to FPR1 specific stimulator fMLF and FPR2, respectively. The stimulator WKYMVm induces the human neutrophil superoxide radical production inhibition activity evaluation comparison results; Table 6 compounds 3 (HCH6-1), compound 6 (HCH30-2), compound 19a (HCH108-4), Compound 19b (HCH108-3), Compound 22a (HCH113-4), Compound 22b (HCH113-3), Compound 24a (HCH99-2), Compound 24b (HCH99-1), Compound 27a (HCH90-2-2) and Compound 27b (HCH90-2-1) was evaluated for the release inhibitory activity of human neutrophil elastase induced by the FPR1 specific stimulator fMLF and the FPR2 specific stimulator WKYMVm, respectively. The IC _{50 in} Tables 5 and 6 is expressed as mean ± standard error (n = 4 or 8), *** p < 0.001 (compared to the control group), and Inh% is the percentage of inhibition below 10 μM.

表五 Table 5

在表五和表六之中，利用fMLF(FPR1活化劑)刺激嗜中性白血球，可得知一系列的雙胜肽衍生物具有抑制人類嗜中性白血球所釋放的超氧自由基及彈性蛋白酶，接著再利用WKYMVm(FPR1/2活化劑)刺激細胞，發現抑制效果減弱，觀察到這些雙胜肽衍生物有選擇性的抑制fMLF所誘導活化的嗜中性白血球。可以從表五和表六觀察到HCH99-2(化合物24a)與HCH6-1(化合物3)抑制效果極佳，IC₅₀分別為0.19±0.07μM和0.32±0.03μM，針對FPR1及FPR1/2選擇性而言，化合物24a相差8倍，化合物3則相差15倍。 In Tables 5 and 6, using fMLF (FPR1 activator) to stimulate neutrophils, it is known that a series of double peptide derivatives have superoxide free radicals and elastases that inhibit the release of human neutrophils. Then, WKYMVm (FPR1/2 activator) was used to stimulate the cells, and it was found that the inhibitory effect was weakened, and it was observed that these double peptide derivatives selectively inhibited the activation of neutrophils by fMLF. It can be observed from Tables 5 and 6 that HCH99-2 ( Compound 24a ) and HCH6-1 ( Compound 3 ) have excellent inhibitory effects with IC ₅₀ of 0.19±0.07 μM and 0.32±0.03 μM, respectively, for FPR1 and FPR1/2 selection. Sexually, compound 24a differs by a factor of eight and compound 3 differs by a factor of 15.

以下實施例一~實施例五係本案之雙胜肽類衍生物的製備方法。而實施例六~實施例十三則是針對各種雙胜肽類衍生物進行以下各項活性測試，比如由FPR1專一性刺激劑fMLF與FPR2專一性刺激劑甲醯肽受體(Formyl Peptide Receptor-Like 1 Agonist，WKYMVm)所誘導的人類嗜中性白血球彈性蛋白酶(neutrophil elastase)的釋放與超氧自由基(superoxide anion)的生成抑制活性評估，與其他相關藥理基轉探討等，有關各項活性測試之方法與實驗步驟，及其活性測試結果如下： The following Example 1 to Example 5 are the preparation methods of the double peptide derivative of the present invention. In the sixth embodiment to the thirteenth embodiment, the following activity tests are performed on various dipeptide derivatives, such as the FPR1 specific stimulator fMLF and the FPR2 specific stimulant meglumine receptor (Formyl Peptide Receptor- Like 1 Agonist, WKYMVm) The release of human neutrophil elastase and the inhibition of superoxide anion production, and other related pharmacological transformations, etc. The test method and experimental procedure, and the activity test results are as follows:

【實施例一】N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters，3-7)以及其相關衍生物(化合物8-10、11a及11b)的製備方法： [Example A] N - (N - aralkyl acyl -L- tryptophyl) -D- phenylalanine methyl ester (N - (N -aroyl-L -tryptophanyl) -D-phenylalanine methyl esters, 3 -7 ) and preparation methods of related derivatives ( compounds 8-10 , 11a and 11b ):

將1.0當量(equiv)左旋色胺酸(L-tryptophan)或左旋5-羥基色胺酸(L-5-hydroxy-tryptophan)(1.0當量)溶於2N氫氧化鈉(NaOH)溶液中並將反應瓶置於冰浴上，採緩慢滴加方式加入醯基氯化物(acyl chloride)類化合物(1.1當量)於反應液中，滴加完即移至室溫下進行反應。反應3小時後，加入1N氯化氫(HCl)做pH值調整動作，利用pH試紙測定觀察，將pH值調至1~2。接著以氯仿(CHCl₃)進行分配萃取，取得之氯仿層經減壓濃縮後即可得到氮取代左旋色胺酸(N-substituded-L-tryptophan)中間產物。中間產物(1.0當量)與D-苯丙氨酸甲酯鹽酸鹽(D-phenylalanine methyl ester)(1.0當量)進行反應，以二氯甲烷(dichloromethane)為溶媒，隨後加入HBTU(2.0當量)及DIEA(1.5當量)在室溫下反應6小時，經以溶媒系統正已烷/乙酸乙酯(6：4)或正已烷/丙酮(7：3)分別進行管柱層析純化，即可得到最終產物(化合物3-10、11a及11b)。 1.0 equivalent (equiv) L-tryptophan or L-5-hydroxy-tryptophan (1.0 eq.) was dissolved in 2N sodium hydroxide (NaOH) solution and the reaction was carried out. The bottle was placed on an ice bath, and an acyl chloride compound (1.1 equivalent) was added to the reaction liquid in a slow dropwise manner, and the reaction was carried out by moving to room temperature after the dropwise addition. After reacting for 3 hours, 1N hydrogen chloride (HCl) was added for pH adjustment, and the pH was adjusted to 1 to 2 by measurement using a pH test paper. Then, the mixture was extracted with chloroform (CHCl ₃ ), and the obtained chloroform layer was concentrated under reduced pressure to obtain a nitrogen-substituted intermediate product of N- substituded-L-tryptophan. The intermediate product (1.0 eq.) was reacted with D-phenylalanine methyl ester (1.0 eq.) with dichloromethane as solvent, followed by HBTU (2.0 eq.) and DIEA (1.5 eq.) was reacted at room temperature for 6 hours and purified by column chromatography with solvent system n-hexane/ethyl acetate (6:4) or n-hexane/acetone (7:3). The final product (compounds 3-10 , 11a and 11b ) was obtained.

N-(N-Benzoyl-L-tryptophanyl)-D-phenylanlanine methyl ester(化合物3，又名HCH6-1) N -( N -Benzoyl-L-tryptophanyl)-D-phenylanlanine methyl ester (Compound 3, also known as HCH6-1)

產率52%。白色粉狀固體，熔點：174-176℃。圓二色光譜(c 0.11mM，MeOH)nm(mdeg)：261(-0.43)，253(-0.40)，239(-0.01)，223(0.46)nm。 The yield was 52%. White powdery solid, melting point: 174-176 °C. Circular dichroism ( c 0.11 mM, MeOH) nm (mdeg): 261 (-0.43), 253 (-.40), 239 (-0.01), 223 (0.46) nm.

¹H NMR(C₅D₅N)：δ 11.79(1H,s,NH),9.41(1H,d,J=8.0Hz, C₁ONH),9.06(1H,d,J=8.0Hz,NH),8.06(2H,d,J=7.2Hz),7.88(1H,d,J=8.0Hz),7.54(1H,d,J=8.0Hz),7.36-7.12(11H,m),5.67(1H,ddd,J=14.4,7.2,7.2Hz),5.23(1H,ddd,J=14.0,7.2,7.2Hz),3.77(1H,dd,J=14.6,7.2Hz),3.63(1H,dd,J=14.4,6.8Hz),3.50(3H,s),3.20(1H,dd,J=13.6,7.2Hz),3.09(1H,dd,J=13.6,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.79 (1H, s, NH), 9.41 (1H, d, J = 8.0 Hz, C ₁ ONH), 9.06 (1H, d, J = 8.0 Hz, NH) , 8.86 (2H, d, J = 7.2 Hz), 7.88 (1H, d, J = 8.0 Hz), 7.54 (1H, d, J = 8.0 Hz), 7.36-7.12 (11H, m), 5.67 (1H, Ddd, J = 14.4, 7.2, 7.2 Hz), 5.23 (1H, ddd, J = 14.0, 7.2, 7.2 Hz), 3.77 (1H, dd, J = 14.6, 7.2 Hz), 3.63 (1H, dd, J = 14.4, 6.8 Hz), 3.50 (3H, s), 3.20 (1H, dd, J = 13.6, 7.2 Hz), 3.09 (1H, dd, J = 13.6, 7.2 Hz).

¹³C NMR(C₅D₅N)： 172.9(s),172.5(s),168.0(s),137.7(3×C,s),131.6(d),130.0(2×C,d),129.0(2×C,d),128.9(s),128.7(2×C,d),128.2(2×C,d),127.3(d),124.6(d),122.0(d),119.5(2×C,d),112.2(d),111.4(s),55.5(d),54.6(d),52.0(q),38.2(t),29.20(t)。 ¹³ C NMR (C ₅ D ₅ N): 172.9(s), 172.5(s), 168.0(s), 137.7(3×C,s), 131.6(d), 130.0(2×C,d),129.0(2×C,d),128.9(s ), 128.7 (2 × C, d), 128.2 (2 × C, d), 127.3 (d), 124.6 (d), 122.0 (d), 119.5 (2 × C, d), 112.2 (d), 111.4 (s), 55.5(d), 54.6(d), 52.0(q), 38.2(t), 29.20(t).

低解析噴灑式質譜(m/z，%)：492[M+Na]⁺(100),470[M+1]⁺(44)。高解析度質譜：m/z 492.1901[M+Na]⁺(計算值；C₂₈H₂₇N₃O₄Na 492.1899)。 Low analytical spray mass spectrometry ( m/z , %): 492 [M+Na] ⁺ (100), 470 [M+1] ⁺ (44). High Resolution Mass Spectrum: m / z 492.1901 [M + Na] + ( _{_{_{calcd; C 28 H 27 N 3 O}}} 4 Na 492.1899).

N-(N-4-氟基苯甲醯基-L-色氨醯基)-D-苯丙氨酸甲酯 (N-(N-4-Fluorobenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物4) N- ( N -4-fluorobenzhydryl -L-chromoylmethyl)-D-phenylalanine methyl ester (N-(N-4-Fluorobenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester , compound 4)

產率31%。白色，粉狀固體，熔點：191-193℃。 The yield was 31%. White, powdery solid, melting point: 191-193 °C.

¹H NMR(C₅D₅N)：δ 11.81(1H,s),9.48(1H,d,J=8.0Hz),9.11(1H,d,J=8.0Hz),8.04(2H,dd,J=8.0Hz),7.87(1H,d,J=7.6Hz),7.52(1H,d J=8.0Hz,H-7),7.32(1H,s),7.26-7.13(7H,m),7.02(2H,t,J=8.6Hz),5.66(1H,ddd,J=14.6,7.2,7.2Hz),5.24(1H,ddd,J=14.2,7.2,7.2Hz),3.75(1H,dd,J=14.4,6.8Hz),3.61(1H,dd,J=14.4,6.8Hz),3.51(3H,s),.3.20(1H,dd,J=14.2,7.2Hz),3.09(1H,dd,J=14.2,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.81 (1H, s), 9.48 (1H, d, J = 8.0 Hz), 9.11 (1H, d, J = 8.0 Hz), 8.04 (2H, dd, J = 8.0 Hz), 7.87 (1H, d, J = 7.6 Hz), 7.52 (1H, d J = 8.0 Hz, H-7), 7.32 (1H, s), 7.26-7.13 (7H, m), 7.02 ( 2H, t, J = 8.6 Hz), 5.66 (1H, ddd, J = 14.6, 7.2, 7.2 Hz), 5.24 (1H, ddd, J = 14.2, 7.2, 7.2 Hz), 3.75 (1H, dd, J = 14.4, 6.8 Hz), 3.61 (1H, dd, J = 14.4, 6.8 Hz), 3.51 (3H, s), .3.20 (1H, dd, J = 14.2, 7.2 Hz), 3.09 (1H, dd, J = 14.2, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ174.4(s),174.0(s),168.4(s),166.4(s,J _C-F=248.0Hz),139.2(2×C,s),133.5(s),132.2(2×C,d,J _C-F=9.0Hz),131.5(2×C,d),130.5(2×C,d),130.4(s),128.8(d),126.1(d),123.5(d),121.0(2×C,d),117.0(2×C,d,J _C-F=22.0Hz),113.7(d),112.9(s),57.1(d),56.1(d),53.6(q),39.7(t),30.7(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ174.4 (s), 174.0 (s), 168.4 (s), 166.4 (s, J CF = 248.0Hz), 139.2 (2 × C, s), 133.5 ( s), 132.2 (2 × C, d, J _CF = 9.0 Hz), 131.5 (2 × C, d), 130.5 (2 × C, d), 130.4 (s), 128.8 (d), 126.1 (d) , 123.5(d), 121.0 (2 × C, d), 117.0 (2 × C, d, J _CF = 22.0 Hz), 113.7 (d), 112.9 (s), 57.1 (d), 56.1 (d), 53.6 (q), 39.7 (t), 30.7 (t).

低解析噴灑式質譜(m/z，%)：510[M+Na]⁺(100),488[M+1]⁺ (37)。高解析度質譜：m/z 510.1802[M+Na]⁺(calcd for C₂₈H₂₆FN₃O₄Na 510.1805)。 Low analytical spray mass spectrometry ( m/z , %): 510 [M+Na] ⁺ (100), 488 [M+1] ⁺ (37). High resolution mass spectrum: m/z 510.1802 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ FN ₃ O ₄ Na 510.1 805).

N-(N-4-氯基苯甲醯基-L-色氨醯基)-D-苯丙氨酸甲酯 (N-(N-4-Chlorobenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物5) N- ( N -4 -Chlorobenzoyl-L-tryptophanyl-D-phenylalanine methyl ester , compound 5)

產率23%。白色粉狀固體，熔點：191-193℃。 The yield was 23%. White powdery solid, melting point: 191-193 ° C.

¹H NMR(C₅D₅N)：δ 11.82(1H,s),9.48(1H,d,J=7.6Hz),9.18(1H,d,J=8.0Hz),7.96(2H,d,J=8.4),7.85(1H,d,J=8.0Hz),7.51(1H,d,J=8.0Hz),7.28(1H,s),7.26-7.12(9H,m),5.65(1H,ddd,J=14.6,6.8,6.8Hz),5.24(1H,ddd,J=14.0,7.6,7.6Hz),3.74(1H,dd,J=14.4,6.8Hz),3.59(1H,dd,J=14.2,7.2Hz),3.50(3H,s),3.20(1H,dd,J=13.8,7.6Hz),3.08(1H,dd,J=13.6,7.6Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.82 (1H, s), 9.48 (1H, d, J = 7.6 Hz), 9.18 (1H, d, J = 8.0 Hz), 7.96 (2H, d, J =8.4), 7.85 (1H, d, J = 8.0 Hz), 7.51 (1H, d, J = 8.0 Hz), 7.28 (1H, s), 7.26-7.12 (9H, m), 5.65 (1H, ddd, J = 14.6, 6.8, 6.8 Hz), 5.24 (1H, ddd, J = 14.0, 7.6, 7.6 Hz), 3.74 (1H, dd, J = 14.4, 6.8 Hz), 3.59 (1H, dd, J = 14.2, 7.2 Hz), 3.50 (3H, s), 3.20 (1H, dd, J = 13.8, 7.6 Hz), 3.08 (1H, dd, J = 13.6, 7.6 Hz).

¹³C NMR(C₅D₅N)：δ 172.8(s),172.5(s),166.9(s),137.7(s),137.3(s),135.7(s),134.2(s),130.0(2×C,d),129.9(2×C,d),129.0(2×C,d),128.9(s),128.8(2×C,d),127.3(d),124.5(d),122.0(d),119.5(2×C,d),112.2(d),111.3(s),55.6(d),54.6(d),52.0(q),38.2(t),29.1(t)。’ ¹³ C NMR (C ₅ D ₅ N): δ 172.8 (s), 172.5 (s), 166.9 (s), 137.7 (s), 137.3 (s), 135.7 (s), 134.2 (s), 130.0 (2) ×C, d), 129.9 (2 × C, d), 129.0 (2 × C, d), 128.9 (s), 128.8 (2 × C, d), 127.3 (d), 124.5 (d), 122.0 ( d), 119.5 (2 x C, d), 112.2 (d), 111.3 (s), 55.6 (d), 54.6 (d), 52.0 (q), 38.2 (t), 29.1 (t). '

低解析噴灑式質譜(m/z，%)：526[M+Na]⁺(100),504[M+1]⁺(33)。高解析度質譜：m/z 526.1506[M+Na]⁺(calcd for C₂₈H₂₆ClN₃O₄Na 526.1509)。 Low analytical spray mass spectrometry ( m/z , %): 526 [M+Na] ⁺ (100), 504 [M+1] ⁺ (33). High-resolution mass spectrum: m/z 526.1506 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ ClN ₃ O ₄ Na 526.1509).

N-(N-4-甲基苯甲醯基-L-色氨醯基)-D-苯丙氨酸甲酯 (N-(N-4-Methylbenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物6) N- ( N -4-methylbenzhydryl -L-chromomethoxymethyl)-D-phenylalanine methyl ester (N-(N-4-Methylbenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester , compound 6)

產率39%。白色粉狀固體，熔點：169-171℃。 The yield was 39%. White powdery solid, melting point: 169-171 °C.

¹H NMR(C₅D₅N)：δ11.77(1H,s),9.41(1H,d,J=8.0Hz),8.98(1H,d,J=8.0Hz),8.00(2H,d,J=8.0Hz),7.89(1H,d,J=7.6Hz),7.54(1H,d,J=8.4Hz),7.33(1H,s),7.30-7.14(7H,m),7.05(2H,d,J=7.6Hz),5.69(1H,ddd,J=14.4,6.8,6.8Hz),5.23(1H,ddd,J=14.0,7.2,7.2Hz),3.79(1H,dd,J=14.6,6.8Hz),3.65(1H,dd,J=14.4,6.8Hz),3.51(3H,s),3.21(1H,dd,J=13.8, 6.8Hz),3.09(1H,dd,J=13.6,7.2Hz),2,13(3H,s)。 ¹ H NMR (C ₅ D ₅ N): δ 11.77 (1H, s), 9.41 (1H, d, J = 8.0 Hz), 8.98 (1H, d, J = 8.0 Hz), 8.00 (2H, d, J = 8.0 Hz), 7.89 (1H, d, J = 7.6 Hz), 7.54 (1H, d, J = 8.4 Hz), 7.33 (1H, s), 7.30-7.14 (7H, m), 7.05 (2H, d, J = 7.6 Hz), 5.69 (1H, ddd, J = 14.4, 6.8, 6.8 Hz), 5.23 (1H, ddd, J = 14.0, 7.2, 7.2 Hz), 3.79 (1H, dd, J = 14.6, 6.8 Hz), 3.65 (1H, dd, J = 14.4, 6.8 Hz), 3.51 (3H, s), 3.21 (1H, dd, J = 13.8, 6.8 Hz), 3.09 (1H, dd, J = =13.6, 7.2 Hz), 2, 13 (3H, s).

¹³C NMR(C₅D₅N)：δ 174.5(s),174.0(s),169.5(s),143.4(s),139.2(2×C,s),134.3(s),131.5(2×C,d),130.9(2×C,d),130.5(2×C,d),130.4(s),129.8(2×C,d),128.8(d),126.1(d),123.5(d),121.0(2×C,d),113.7(d),112.9(s),57.0(d),56.2(d),53.6(q),39.7(t),30.7(t),22.9(q)。 ¹³ C NMR (C ₅ D ₅ N): δ 174.5 (s), 174.0 (s), 169.5 (s), 143.4 (s), 139.2 (2 × C, s), 134.3 (s), 131.5 (2 × C, d), 130.9 (2 × C, d), 130.5 (2 × C, d), 130.4 (s), 129.8 (2 × C, d), 128.8 (d), 126.1 (d), 123.5 (d ), 121.0 (2 × C, d), 113.7 (d), 112.9 (s), 57.0 (d), 56.2 (d), 53.6 (q), 39.7 (t), 30.7 (t), 22.9 (q) .

低解析噴灑式質譜(m/z，%)：506[M+Na]⁺(99),484[M+1]⁺(100)。高解析度質譜：m/z 506.2059[M+Na]⁺(calcd for C₂₉H₂₉N₃O₄Na 506.2056)。 Low analytical spray mass spectrometry ( m/z , %): 506 [M+Na] ⁺ (99), 484 [M+1] ⁺ (100). High-resolution mass spectrum: m/z 506.2059 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₄ Na 506.2056).

N-(N-4-甲氧基苯甲醯基-L-色氨醯基)-D-苯丙氨酸甲酯 (N-(N-4-Methoxylbenzoyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物7) N- ( N -4-methoxybenzimidyl -L-chromomethoxymethyl)-D-phenylalanine methyl (N-(N-4-Methoxylbenzoyl-L-tryptophanyl)-D-phenylalanine methyl Ester , compound 7)

產率47%。白色粉狀固體，熔點：151-153℃。 The yield was 47%. White powdery solid, melting point: 151-153 ° C.

¹H NMR(C₅D₅N)：δ11.80(1H,s),9.39(1H,d,J=8.0Hz),8.92(1H,d,J=8.0Hz),8.09(2H,d,J=8.4Hz),7.99(1H,d,J=8.0Hz),7.53(1H,d,J=8.0Hz),7.33(1H,s),7.33-7.13(7H,m),6.88(2H,d,J=8.4Hz),5.69(1H,ddd,J=14.4,6.8,6.8Hz),5.23(1H,ddd,J=14.0,7.2,7.2Hz),3.78(1H,dd,J=14.2,6.8Hz),3.64(1H,dd,J=14.6,6.8Hz),3.59(3H,s),3.50(3H,s),3.20(1H,dd,J=13.6,7.2Hz),3.09(1H,dd,J=13.6,7.6Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.80 (1H, s), 9.39 (1H, d, J = 8.0 Hz), 8.92 (1H, d, J = 8.0 Hz), 8.09 (2H, d, J = 8.4 Hz), 7.99 (1H, d, J = 8.0 Hz), 7.53 (1H, d, J = 8.0 Hz), 7.33 (1H, s), 7.33 - 7.13 (7H, m), 6.88 (2H, d, J = 8.4 Hz), 5.69 (1H, ddd, J = 14.4, 6.8, 6.8 Hz), 5.23 (1H, ddd, J = 14.0, 7.2, 7.2 Hz), 3.78 (1H, dd, J = 14.2, 6.8 Hz), 3.64 (1H, dd, J = 14.6, 6.8 Hz), 3.59 (3H, s), 3.50 (3H, s), 3.20 (1H, dd, J = 13.6, 7.2 Hz), 3.09 (1H, Dd, J = 13.6, 7.6 Hz).

¹³C NMR(C₅D₅N)：δ174.6(s),174.0(s),169.0(s),164.1(s),139.2(2×C,s),131.6(2×C,d),131.5(2×C,d),130.5(2×C,d),130.4(s),129.4(s),128.8(d),126.1(d),123.5(d),121.0(2×C,d),115.6(2×C,d),113.7(d),113.0(s),57.0(q),56.9(d),56.1(d),53.6(q),39.03(t),30.7(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ174.6 (s), 174.0 (s), 169.0 (s), 164.1 (s), 139.2 (2 × C, s), 131.6 (2 × C, d) , 131.5 (2 × C, d), 130.5 (2 × C, d), 130.4 (s), 129.4 (s), 128.8 (d), 126.1 (d), 123.5 (d), 121.0 (2 × C, d), 115.6 (2 × C, d), 113.7 (d), 113.0 (s), 57.0 (q), 56.9 (d), 56.1 (d), 53.6 (q), 39.03 (t), 30.7 (t ).

低解析噴灑式質譜(m/z，%)：522[M+Na]⁺(100)，500[M+1]⁺(38)。高解析度質譜：m/z 522.2007[M+Na]⁺(calcd for C₂₉H₂₉N₃O₅Na 522.2005)。 Low analytical spray mass spectrometry ( m/z , %): 522 [M+Na] ⁺ (100), 500 [M+1] ⁺ (38). High resolution mass spectrum: m/z 522.2007 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₅ Na 522.2005).

N-(N-苯乙醯基-L-色氨醯基)-D-苯丙氨酸甲酯 (N-(N-Phenylacetyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物8) N - (N - phenyl acetyl group -L- tryptophyl acyl) -D- phenylalanine methyl ester (N- (N-Phenylacetyl-L -tryptophanyl) -D-phenylalanine methyl ester, Compound 8)

產率43%。白色粉狀固體，熔點：182-184℃。 The yield was 43%. White powdery solid, melting point: 182-184 °C.

¹H NMR(C₅D₅N)：δ11.76(1H,s),9.34(1H,d,J=8.0Hz),8.98(1H,d,J=8.4Hz),7.81(1H,d,J=8.0Hz),7.53(1H,d,J=8.4Hz),7.33(2H,d,J=7.2Hz),7.27-7.13(11H,m),5.50(1H,ddd,J=14.6,6.8,6.8Hz),5.19(1H,ddd,J=14.0,7.2,7.2Hz),3.75-3.46(4H,m),3.49(3H,s),3.17(1H,dd,J=13.6,6.8Hz),3.05(1H,dd,J=13.8,6.8Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.76 (1H, s), 9.34 (1H, d, J = 8.0 Hz), 8.98 (1H, d, J = 8.4 Hz), 7.81 (1H, d, J = 8.0 Hz), 7.53 (1H, d, J = 8.4 Hz), 7.33 (2H, d, J = 7.2 Hz), 7.27-7.13 (11H, m), 5.50 (1H, ddd, J = 14.6, 6.8 , 6.8 Hz), 5.19 (1H, ddd, J = 14.0, 7.2, 7.2 Hz), 3.75-3.46 (4H, m), 3.49 (3H, s), 3.17 (1H, dd, J = 13.6, 6.8 Hz) , 3.05 (1H, dd, J = 13.8, 6.8 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),172.5(s),171.1(s),137.7(s),137.6(s),136.9(s),130.0(2×C,d),129.9(2×C,d),129.0(2×C,d),128.9(s),128.9(2×C,d),127.3(d),127.0(d),124.5(d),121.9(d),119.5(d),119.4(d),112.1(d),111.2(s),54.9(d),54.6(d),52.0(q),43.5(t),38.2(t),29.3(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.8 (s), 172.5 (s), 171.1 (s), 137.7 (s), 137.6 (s), 136.9 (s), 130.0 (2 x C, d ), 129.9 (2 × C, d), 129.0 (2 × C, d), 128.9 (s), 128.9 (2 × C, d), 127.3 (d), 127.0 (d), 124.5 (d), 121.9 (d), 119.5(d), 119.4(d), 112.1(d), 111.2(s), 54.9(d), 54.6(d), 52.0(q), 43.5(t), 38.2(t), 29.3 (t).

低解析噴灑式質譜(m/z，%)：506[M+Na]⁺(100),484[M+1]⁺(87)。高解析度質譜：m/z 506.2060[M+Na]⁺(calcd for C₂₉H₂₉N₃O₄Na 506.2056)。 Low analytical spray mass spectrometry ( m/z , %): 506 [M+Na] ⁺ (100), 484 [M+1] ⁺ (87). High-resolution mass spectrum: m/z 506.2060 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₄ Na 506.2056).

N-(N-苄基-丙醯基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-Phenylpropanoyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物9) N - (N - benzyl - propan-acyl amino acyl -L- color) -D- phenylalanine methyl ester (N- (N-Phenylpropanoyl-L -tryptophanyl) -D-phenylalanine methyl ester, Compound 9)

產率24%。白色粉狀固體，熔點：159-161℃。 The yield was 24%. White powdery solid, melting point: 159-161 ° C.

¹H NMR(C₅D₅N)：δ11.78(1H,s),9.27(1H,d,J=8.0Hz),8.88(1H,d,J=8.4Hz),7.85(1H,d,J=7.6Hz),7.55(1H,d,J=7.6Hz),7.27-7.12(13H,m),5.52(1H,ddd,J=14.8,7.2,7.2Hz),5.21(1H,ddd,J=14.0,7.2,7.2Hz),3.65(1H,dd.J=14.4,6.8Hz),3.50(3H,s),3.50-3.45(1H,m),3.17(1H,J=13.6,6.8Hz),3.09-2.97(3H,m),2.67-2.61(2H,m)。 ¹ H NMR (C ₅ D ₅ N): δ 11.78 (1H, s), 9.27 (1H, d, J = 8.0 Hz), 8.88 (1H, d, J = 8.4 Hz), 7.85 (1H, d, J = 7.6 Hz), 7.55 (1H, d, J = 7.6 Hz), 7.27-7.12 (13H, m), 5.52 (1H, ddd, J = 11.8, 7.2, 7.2 Hz), 5.21 (1H, ddd, J =14.0, 7.2, 7.2 Hz), 3.65 (1H, dd. J = 14.4, 6.8 Hz), 3.50 (3H, s), 3.50-3.45 (1H, m), 3.17 (1H, J = 13.6, 6.8 Hz) , 3.09-2.97 (3H, m), 2.67-2.61 (2H, m).

¹³C NMR(C₅D₅N)：δ174.4(s),174.0(s),173.9(s),143.7(s),139.2(2×C,s),131.5(2×C,d),130.5(2×C,d),130.4(d),130.4(d),130.3(2×C,d),128.8(s),128.0(d),126.0(d),123.4(d),121.0(d),120.9(d),113.6(d),112.9(s),56.4(d),56.0(d),53.5(q),39.7(2×C,t),33.7(t),30.8(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 174.4 (s), 174.0 (s), 173.9 (s), 143.7 (s), 139.2 (2 x C, s), 131.5 (2 x C, d) , 130.5 (2 × C, d), 130.4 (d), 130.4 (d), 130.3 (2 × C, d), 128.8 (s), 128.0 (d), 126.0 (d), 123.4 (d), 121.0 (d), 120.9(d), 113.6(d), 112.9(s), 56.4(d), 56.0(d), 53.5(q), 39.7(2xC,t),33.7(t),30.8( t).

低解析噴灑式質譜(m/z，%)：520[M+Na]⁺(100),498[M+1]⁺(39)。高解析度質譜：m/z 520.2215[M+Na]⁺(Calcd for C₂₉H₂₉N₃O₄Na 520.2212)。 Low analytical spray mass spectrometry ( m/z , %): 520 [M+Na] ⁺ (100), 498 [M+1] ⁺ (39). High-resolution mass spectrum: m/z 520.2215 [M+Na] ⁺ (Calcd for C ₂₉ H ₂₉ N ₃ O ₄ Na 520.2212).

N-(N-苄基-碳醯基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-Benzyloxycarbonyl-L-tryptophanyl)-D-phenylalanine methyl ester ，化合物10) N - (N - benzyl - carbon acyl -L- tryptophyl acyl) -D- phenylalanine methyl ester (N- (N-Benzyloxycarbonyl-L -tryptophanyl) -D-phenylalanine methyl ester, Compound 10)

產率67%。白色粉狀固體，熔點：147-149℃。 The yield was 67%. White powdery solid, melting point: 147-149 ° C.

¹H NMR(C₅D₅N)：δ11.76(1H,s),9.32(1H,d,J=7.6Hz),8.63(1H,d,J=8.4Hz),7.88(1H,d,J=7.6Hz),7.54(1H,d,J=8.4Hz),7.32-7.14(13H,m),5.25-5.13(4H,m),3.72(1H,dd,J=14.4,7.2Hz),3.55(1H,dd,J=14.8,7.2Hz),3.49(3H,s),3.17(1H,dd,J=13.6,7.2Hz),3.07(1H,dd,J=13.8,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.76 (1H, s), 9.32 (1H, d, J = 7.6 Hz), 8.63 (1H, d, J = 8.4 Hz), 7.88 (1H, d, J = 7.6 Hz), 7.54 (1H, d, J = 8.4 Hz), 7.32 - 7.14 (13H, m), 5.25 - 5.13 (4H, m), 3.72 (1H, dd, J = 14.4, 7.2 Hz), 3.55 (1H, dd, J = 14.8, 7.2 Hz), 3.49 (3H, s), 3.17 (1H, dd, J = 13.6, 7.2 Hz), 3.07 (1H, dd, J = 13.8, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.9(s),172.5(s),157.2(s),137.9(s),137.7(2×C,s),130.0(2×C,d),129.0(2×C,d),129.0(d),128.9(2×C,d),128.9(s),128.3(2×C,d),127.3(s),124.6(d),122.0(d),119.5(2×C,d),112.2(d),111.3(s),66.6(t),57.0(d),54.5(d),52.0(q),38.2(t),29.7(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.9 (s), 172.5 (s), 157.2 (s), 137.9 (s), 137.7 (2 × C, s), 130.0 (2 × C, d) , 129.0 (2 × C, d), 129.0 (d), 128.9 (2 × C, d), 128.9 (s), 128.3 (2 × C, d), 127.3 (s), 124.6 (d), 122.0 ( d), 119.5 (2 × C, d), 112.2 (d), 111.3 (s), 66.6 (t), 57.0 (d), 54.5 (d), 52.0 (q), 38.2 (t), 29.7 (t ).

低解析噴灑式質譜(m/z，%)：522[M+Na]⁺(100),500[M+1]⁺(30)。高解析度質譜：m/z 522.2008[M+Na]⁺(calcd for C₂₉H₂₉N₃O₅Na 522.2005)。 Low analytical spray mass spectrometry ( m/z , %): 522 [M+Na] ⁺ (100), 500 [M+1] ⁺ (30). High resolution mass spectrum: m/z 522.2008 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₅ Na 522.2005).

N-(N-苄基-5-羥基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-Benzoyl-5-hydroxy-L-tryptophanyl)-D-phenylalanine methyl esters ，化合物11a,11b) N - (N - benzyl-5-hydroxy -L- tryptophyl acyl) -D- phenylalanine methyl ester (N- (N-Benzoyl-5 -hydroxy-L-tryptophanyl) -D-phenylalanine methyl esters , compound 11a, 11b)

化合物11a：產率23%。白色粉狀固體，熔點：112-114℃。 Compound 11a: Yield 23%. White powdery solid, melting point: 112-114 ° C.

¹H NMR(C₅D₅N)：δ11.54(1H,s,NH),9.39(1H,d,J=8.0Hz,NH),9.04(1H,d,J=8.0Hz,NH),8.06(2H,d,J=7.2Hz),7.74(1H,d,J=2.0Hz),7.47(1H,d,J=8.8Hz),7.35(1H,t,J=7.2Hz),7.29-7.13(9H,m),5.65(1H,dd,J=14.2,7.2Hz),5.23(1H,dd,J=14.2,7.2Hz),3.72(1H,dd,J=14.2,7.2Hz),3.59(1H,dd,J=14.2,7.2Hz),3.47(3H,s),3.15(1H,dd,J=13.6,7.2Hz),3.05(1H,dd,J=13.6,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.54 (1H, s, NH), 9.39 (1H, d, J = 8.0 Hz, NH), 9.04 (1H, d, J = 8.0 Hz, NH), 8.06 (2H, d, J = 7.2 Hz), 7.74 (1H, d, J = 2.0 Hz), 7.47 (1H, d, J = 8.8 Hz), 7.35 (1H, t, J = 7.2 Hz), 7.29- 7.13 (9H, m), 5.65 (1H, dd, J = 14.2, 7.2 Hz), 5.23 (1H, dd, J = 14.2, 7.2 Hz), 3.72 (1H, dd, J = 14.2, 7.2 Hz), 3.59 (1H, dd, J = 14.2, 7.2 Hz), 3.47 (3H, s), 3.15 (1H, dd, J = 13.6, 7.2 Hz), 3.05 (1H, dd, J = 13.6, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),172.3(s),167.7(s),152.4(s),137.4(s),135.5(s),132.2(s),131.4(d),129.8(s),129.8(2×C,d),128.8(2×C, d),128.5(2×C,d),128.0(2×C,d),127.1(d),125.0(d),112.8(d),112.6(d),110.3(s),103.9(d),55.5(d),54.5(d),51.8(q),38.0(t),29.2(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.7 (s), 172.3 (s), 167.7 (s), 152.4 (s), 137.4 (s), 135.5 (s), 132.2 (s), 131.4 ( d), 129.8(s), 129.8 (2 x C, d), 128.8 (2 x C, d), 128.5 (2 x C, d), 128.0 (2 x C, d), 127.1 (d), 125.0 (d), 112.8(d), 112.6(d), 110.3(s), 103.9(d), 55.5(d), 54.5(d), 51.8(q), 38.0(t), 29.2(t).

低解析噴灑式質譜(m/z，%)：508[M+Na]⁺(100)。高解析度質譜：m/z 508.1850[M+Na]⁺(calcd for C₂₈H₂₇N₃O₅Na 508.1848)。 Low analytical spray mass spectrometry ( m/z , %): 508 [M+Na] ⁺ (100). High resolution mass spectrum: m/z 508.1850 [M+Na] ⁺ (calcd for C ₂₈ H ₂₇ N ₃ O ₅ Na 508.1848).

化合物11b：產率17%。白色粉狀固體，熔點：110-112℃。 Compound 11b: Yield 17%. White powdery solid, melting point: 110-112 ° C.

¹H NMR(C₅D₅N)：δ11.53(1H,s,NH),9.41(1H,d,J=8.0Hz,NH),9.11(1H,d,J=8.0Hz,NH),8.10(2H,d,J=8.0Hz),7.64(1H,d,J=1.6Hz),7.43(1H,d,J=8.0Hz),7.37(1H,t,J=7.6Hz),7.30(2H,t,J=7.6Hz),7.26(2H,t,J=7.6Hz),7.19-7.13(4H,m),7.08(1H,t,J=8.0Hz),5.65(1H,dd,J=14.4,7.2Hz),5.21(1H,dd,J=14.2,7.2Hz),3.74(1H,dd,J=14.4,7.2Hz),3.61(1H,dd,J=14.4,7.2Hz),3.55(3H,s),3.27(1H,dd,J=13.6,7.2Hz),3.15(1H,dd,J=13.6,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.53 (1H, s, NH), 9.41 (1H, d, J = 8.0 Hz, NH), 9.11 (1H, d, J = 8.0 Hz, NH), 8.10 (2H, d, J = 8.0 Hz), 7.64 (1H, d, J = 1.6 Hz), 7.43 (1H, d, J = 8.0 Hz), 7.37 (1H, t, J = 7.6 Hz), 7.30 ( 2H, t, J = 7.6 Hz), 7.26 (2H, t, J = 7.6 Hz), 7.19-7.13 (4H, m), 7.08 (1H, t, J = 8.0 Hz), 5.65 (1H, dd, J =14.4, 7.2 Hz), 5.21 (1H, dd, J = 14.2, 7.2 Hz), 3.74 (1H, dd, J = 14.4, 7.2 Hz), 3.61 (1H, dd, J = 14.4, 7.2 Hz), 3.55 (3H, s), 3.27 (1H, dd, J = 13.6, 7.2 Hz), 3.15 (1H, dd, J = 13.6, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.6(s),172.3(s),167.8(s),152.3(s),137.5(s),135.5(s),132.2(s),131.4(d),129.8(2×C,d),129.7(s),128.7(2×C,d),128.6(2×C,d),128.1(2×C,d),127.0(d),125.0(d),112.8(d),112.5(d),110.3(s),103.9(d),55.2(d),54.5(d),51.9(q),38.2(t),28.9(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.6 (s), 172.3 (s), 167.8 (s), 152.3 (s), 137.5 (s), 135.5 (s), 132.2 (s), 131.4 ( d), 129.8 (2 × C, d), 129.7 (s), 128.7 (2 × C, d), 128.6 (2 × C, d), 128.1 (2 × C, d), 127.0 (d), 125.0 (d), 112.8(d), 112.5(d), 110.3(s), 103.9(d), 55.2(d), 54.5(d), 51.9(q), 38.2(t), 28.9(t).

【實施例二】N-(N-苄基-L-色氨醯基)-D-苯丙氨醇(N-(N-benzoyl-L-tryptophanyl)-D-phenylalaninol，化合物12a-13a及12b-13b)的製備方法如下： [Second Embodiment] N - (N - benzyl-amino acyl -L- color) -D- phenylalaninol (N- (N-benzoyl-L -tryptophanyl) -D-phenylalaninol, 12a-13a and compound 12b -13b ) is prepared as follows:

N-苯甲醯-左旋色胺酸(N-benzoyl-L-tryptophan)(1.0當量)或N-苯甲醯-5羥基左旋色胺酸(N-benzoyl-5-hydroxy-L-tryptophan)(1.0當量)各別與DL-2-氨基-3-苯基-1-丙醇(DL-2-Amino-3-phenyl-1-propanol)(1.0當量)及D-phenylalanyl-glycine-nitrile(1.0當量)進行反應，以二氯甲烷(dichloromethane)為溶媒，隨後加入HBTU(2.0當量)及DIEA(1.5當量)在室溫下反應6小時，經分別以溶媒系統甲醇/氯仿(1：20)或乙酸乙酯進行管柱層析純化，即可分別含有化合物12a、12b或13a、13b的混合物；將上述混合物以溶媒系統35%乙腈(MeCN)+0.3%三氟乙酸(TFA)進行高效液相色譜法(HPLC)分離，即可得到最終產物(化合物12a、12b及13a、13b)。 N - benzoyl - L-tryptophan (N -benzoyl-L-tryptophan) (1.0 eq) or N - benzoyl -5-hydroxy-L-tryptophan (N -benzoyl-5-hydroxy- L-tryptophan) ( 1.0 equivalent) each with DL-2-amino-3-phenyl-1-propanol (1.0 equivalent) and D-phenylalanyl-glycine-nitrile (1.0) Equivalent) The reaction was carried out with dichloromethane as a solvent, followed by addition of HBTU (2.0 eq.) and DIEA (1.5 eq.) at room temperature for 6 hours, respectively, by solvent system methanol/chloroform (1:20) or The ethyl acetate was purified by column chromatography to contain a mixture of compounds 12a , 12b or 13a, 13b , respectively; the mixture was subjected to high performance liquid phase in a solvent system of 35% acetonitrile (MeCN) + 0.3% trifluoroacetic acid (TFA). The final product ( compounds 12a , 12b and 13a, 13b ) is obtained by chromatography (HPLC).

N-(N-苄基-L-色氨醯基)-5-羥基-D-苯丙氨醇(N-(N-benzoyl-L-tryptophanyl)-5-hydroxy-D-phenylalaninol ，化合物12a和12b) N - (N - benzyl-acyl -L- tryptophyl) - 5 - hydroxy - -D- -phenylalaninol (N- (N-benzoyl-L -tryptophanyl) -5-hydroxy-D-phenylalaninol, and compound 12a 12b)

化合物12a：產率16%。白色粉狀固體，熔點：175-177℃。 Compound 12a: Yield 16%. White powdery solid, melting point: 175-177 °C.

¹H NMR(C₅D₅N)：δ11.77(1H,s,NH),8.97(1H,d,J=8.0Hz,NH),8.88(1H,d,J=8.0Hz,NH),8.05(2H,d,J=7.2Hz),7.89(1H,d,J=8.0Hz),7.51(1H,d,J=8.0Hz),7.36-7.32(4H,m),7.28-7.22(5H,m),7.19(1H,m),7.09(1H,t,J=7.6Hz),5.58(1H,dd,J=14.4,6.8Hz),5.50(1H,br.s,OH),4.75(1H,m),3.91(2H,m),3.75(1H,dd,J=14.4,6.8Hz),3.64(1H,dd,J=14.4,6.8Hz),3.15(1H,dd,J=13.4,7.2Hz),3.03(1H,dd,J=13.4,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.77 (1H, s, NH), 8.97 (1H, d, J = 8.0 Hz, NH), 8.88 (1H, d, J = 8.0 Hz, NH), 8.05 (2H, d, J = 7.2 Hz), 7.89 (1H, d, J = 8.0 Hz), 7.51 (1H, d, J = 8.0 Hz), 7.36-7.32 (4H, m), 7.28-7.22 (5H , m), 7.19 (1H, m), 7.09 (1H, t, J = 7.6 Hz), 5.58 (1H, dd, J = 14.4, 6.8 Hz), 5.50 (1H, br.s, OH), 4.75 ( 1H, m), 3.91 (2H, m), 3.75 (1H, dd, J = 14.4, 6.8 Hz), 3.64 (1H, dd, J = 14.4, 6.8 Hz), 3.15 (1H, dd, J = 13.4, 7.2 Hz), 3.03 (1H, dd, J = 13.4, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),167.9(s),139.8(s),137.7(s),135.7(s),131.5(d),130.2(d),129.0(s),128.9(2×C,d),128.7(2×C,d),128.2(2×C,d),126.7(d),124.6(d),121.9(d),119.5(d),119.4(d),112.1(d),111.5(s),63.2(t),55.9(d),53.8(d),37.7(t),29.4(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.7 (s), 167.9 (s), 139.8 (s), 137.7 (s), 135.7 (s), 131.5 (d), 130.2 (d), 129.0 ( s), 128.9 (2 × C, d), 128.7 (2 × C, d), 128.2 (2 × C, d), 126.7 (d), 124.6 (d), 121.9 (d), 119.5 (d), 119.4(d), 112.1(d), 111.5(s), 63.2(t), 55.9(d), 53.8(d), 37.7(t), 29.4(t).

低解析噴灑式質譜(m/z，%)：464[M+Na]⁺(100),442[M+1]⁺(5)。高解析度質譜：m/z 464.1947[M+Na]⁺(calcd for C₂₇H₂₇N₃O₃Na 464.1950)。 Low analytical spray mass spectrometry ( m/z , %): 464 [M+Na] ⁺ (100), 442 [M+1] ⁺ (5). High-resolution mass spectrum: m/z 464.1947 [M+Na] ⁺ (calcd for C ₂₇ H ₂₇ N ₃ O ₃ Na 464.1950).

化合物12b：產率11%。白色粉狀固體，熔點：173-175℃。 Compound 12b: Yield 11%. White powdery solid, melting point: 173-175 °C.

¹H NMR(C₅D₅N)：δ11.78(1H,s,NH),9.07(1H,d,J=7.6Hz,NH),8.87(1H,d,J=8.0Hz,NH),8.09(2H,d,J=7.6Hz),7.85(1H,d,J=8.0Hz),7.52(1H,br.s),7.48(1H,d,J=8.0Hz),7.29(2H,t,J=7.6Hz),7.27(2H,t,J=7.6Hz),7.22-7.15(3H,m),7.09(2H,t,J=8.6Hz),5.62(1H,dd,J=14.4,7.2Hz),4.71(1H,m),3.87(2H,m),3.80(1H,dd,J=14.4,7.2Hz),3.68(1H,dd,J=14.4,7.2Hz),3.21(1H,dd,J=13.6,6.8Hz),3.09(IH,dd,J=13.6,6.8Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.78 (1H, s, NH), 9.07 (1H, d, J = 7.6 Hz, NH), 8.87 (1H, d, J = 8.0 Hz, NH), 8.09 (2H, d, J = 7.6 Hz), 7.85 (1H, d, J = 8.0 Hz), 7.52 (1H, br.s), 7.48 (1H, d, J = 8.0 Hz), 7.29 (2H, t , J = 7.6 Hz), 7.27 (2H, t, J = 7.6 Hz), 7.22 - 7.15 (3H, m), 7.09 (2H, t, J = 8.6 Hz), 5.62 (1H, dd, J = 14.4, 7.2 Hz), 4.71 (1H, m), 3.87 (2H, m), 3.80 (1H, dd, J = 14.4, 7.2 Hz), 3.68 (1H, dd, J = 14.4, 7.2 Hz), 3.21 (1H, Dd, J = 13.6, 6.8 Hz), 3.09 (IH, dd, J = 13.6, 6.8 Hz).

¹³C NMR(C₅D₅N)：δ172.5(s),167.8(s),139.6(s),137.5(s),135.5(s),131.4(d),129.9(2×C,d),128.7(s),128.6(2×C,d),128.5(2×C,d),128.1(2×C,d),126.4(d),124.4(d),121.7(d),119.3(d),119.2(d),111.9(d),111.4(s),63.1(t),55.5(d),53.8(d),37.6(t),28.8(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.5 (s), 167.8 (s), 139.6 (s), 137.5 (s), 135.5 (s), 131.4 (d), 129.9 (2 x C, d ), 128.7 (s), 128.6 (2 × C, d), 128.5 (2 × C, d), 128.1 (2 × C, d), 126.4 (d), 124.4 (d), 121.7 (d), 119.3 (d), 119.2(d), 111.9(d), 111.4(s), 63.1(t), 55.5(d), 53.8(d), 37.6(t), 28.8(t).

低解析噴灑式質譜(m/z，%)：464[M+Na]⁺(100),442[M+1]⁺(37)。高解析度質譜：m/z 464.1947[M+Na]⁺(calcd for C₂₇H₂₇N₃O₃Na 464.1950)。 Low analytical spray mass spectrometry ( m/z , %): 464 [M+Na] ⁺ (100), 442 [M+1] ⁺ (37). High-resolution mass spectrum: m/z 464.1947 [M+Na] ⁺ (calcd for C ₂₇ H ₂₇ N ₃ O ₃ Na 464.1950).

N-(N-苄基-L-色氨醯基)-D-苯丙氨醇(N-(N-benzoyl-L-tryptophanyl)-D-phenylalaninol ，化合物13a和13b) N - (N - benzyl-amino acyl -L- color) -D- phenylalaninol (N- (N-benzoyl-L -tryptophanyl) -D-phenylalaninol, 13a and compound 13b)

化合物13a：產率53%。白色粉狀固體，熔點：151-153℃。 Compound 13a: Yield 53%. White powdery solid, melting point: 151-153 ° C.

¹H NMR(C₅D₅N)：δ11.53(1H,s,NH),8.96(1H,d,J=7.6Hz,NH),8.82(1H,d,J=8.0Hz,NH),8.07(2H,d,J=7.6Hz),7.76(1H,s),7.46(1H,d,J=8.4Hz),7.34-7.13(10H,m),5.95(1H,br.s,OH),5.56(1H,dd,J=14.0,7.2Hz),4.73(1H,m),3.91(1H,dd,J=10.8,5.2Hz),3.85(1H,dd,J=10.8,5.2Hz),3.70(1H,dd,J=14.0,7.2Hz),3.60(1H,dd,J=14.0,7.2Hz),3.11(1H,dd,J=13.6,7.2Hz),3.00(1H,dd,J=13.6,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.53 (1H, s, NH), 8.96 (1H, d, J = 7.6 Hz, NH), 8.82 (1H, d, J = 8.0 Hz, NH), 8.07 (2H, d, J = 7.6 Hz), 7.76 (1H, s), 7.46 (1H, d, J = 8.4 Hz), 7.34 - 7.13 (10H, m), 5.95 (1H, br.s, OH) , 5.56 (1H, dd, J = 14.0, 7.2 Hz), 4.73 (1H, m), 3.91 (1H, dd, J = 10.8, 5.2 Hz), 3.85 (1H, dd, J = 10.8, 5.2 Hz), 3.70 (1H, dd, J = 14.0, 7.2 Hz), 3.60 (1H, dd, J = 14.0, 7.2 Hz), 3.11 (1H, dd, J = 13.6, 7.2 Hz), 3.00 (1H, dd, J = 13.6, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.5(s),167.6(s),152.4(s),139.5(s),135.5(s),132.2(s),131.4(d),130.1(2×C,d),129.8(s),128.7(2×C,d),128.5(2×C,d),128.0(2×C,d),125.0(d),112.8(d),112.5(d),110.5(s),104.0(d),62.8(t),55.8(d),53.6(d),37.5(t),29.5(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.5 (s), 167.6 (s), 152.4 (s), 139.5 (s), 135.5 (s), 132.2 (s), 131.4 (d), 130.1 ( 2 × C, d), 129.8 (s), 128.7 (2 × C, d), 128.5 (2 × C, d), 128.0 (2 × C, d), 125.0 (d), 112.8 (d), 112.5 (d), 110.5 (s), 104.0 (d), 62.8 (t), 55.8 (d), 53.6 (d), 37.5 (t), 29.5 (t).

低解析噴灑式質譜(m/z，%)：480[M+Na]⁺(100),458[M+1]⁺(18)。高解析度質譜：m/z 480.1897[M+Na]⁺(calcd for C₂₇H₂₇N₃O₄Na 480.1899)。 Low analytical spray mass spectrometry ( m/z , %): 480 [M+Na] ⁺ (100), 458 [M+1] ⁺ (18). High-resolution mass spectrum: m/z 480.1897 [M+Na] ⁺ (calcd for C ₂₇ H ₂₇ N ₃ O ₄ Na 480.1899).

化合物13b：產率25%。白色粉狀固體，熔點：120-122℃。 Compound 13b: Yield 25%. White powdery solid, melting point: 120-122 ° C.

¹H NMR(C₅D₅N)：δ11.53(1H,s,NH),9.05(1H,d,J=8.0Hz,NH),8.76(1H,d,J=8.0Hz,NH),8.10(2H,d,J=7.6Hz),7.71(1H,s),7.55(1H,s),7.46(1H,d,J=8.0Hz),7.38-7.35(3H,m),7.30(2H,t,J=7.6Hz),7.18-7.14(4H,m),7.08(1H,d,J=8.0Hz),5.90(1H,br.s,OH),5.59(1H,dd,J= 14.2,7.2Hz),4.69(1H,m),3.88(1H,dd,J=10.4,5.2Hz),3.82(1H,dd,J=10.4,5.2Hz),3.74(1H,dd,J=14.2,7.2Hz),3.64(1H,dd,J=14.2,7.2Hz),3.20(1H,dd,J=13.6,7.2Hz),3.07(1H,dd,J=13.6,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.53 (1H, s, NH), 9.05 (1H, d, J = 8.0 Hz, NH), 8.76 (1H, d, J = 8.0 Hz, NH), 8.10 (2H, d, J = 7.6 Hz), 7.71 (1H, s), 7.55 (1H, s), 7.46 (1H, d, J = 8.0 Hz), 7.38-7.35 (3H, m), 7.30 (2H) , t, J = 7.6 Hz), 7.18-7.14 (4H, m), 7.08 (1H, d, J = 8.0 Hz), 5.90 (1H, br.s, OH), 5.59 (1H, dd, J = 14.2) , 7.2 Hz), 4.69 (1H, m), 3.88 (1H, dd, J = 10.4, 5.2 Hz), 3.82 (1H, dd, J = 10.4, 5.2 Hz), 3.74 (1H, dd, J = 14.2, 7.2 Hz), 3.64 (1H, dd, J = 14.2, 7.2 Hz), 3.20 (1H, dd, J = 13.6, 7.2 Hz), 3.07 (1H, dd, J = 13.6, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.4(s),167.7(s),152.3(s),139.6(s),135.5(s),132.2(s),131.4(d),129.9(2×C,d),129.8(s),128.6(2×C,d),128.5(2×C,d),128.1(2×C,d),126.4(d),125.0(d),112.8(d),112.5(d),110.6(s),104.0(d),63.8(t),55.5(d),53.7(d),37.6(t),28.9(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.4 (s), 167.7 (s), 152.3 (s), 139.6 (s), 135.5 (s), 132.2 (s), 131.4 (d), 129.9 ( 2 × C, d), 129.8 (s), 128.6 (2 × C, d), 128.5 (2 × C, d), 128.1 (2 × C, d), 126.4 (d), 125.0 (d), 112.8 (d), 112.5(d), 110.6(s), 104.0(d), 63.8(t), 55.5(d), 53.7(d), 37.6(t), 28.9(t).

低解析噴灑式質譜(m/z，%)：480[M+Na]⁺(100),458[M+1]⁺(82)。高解析度質譜：m/z 480.1897[M+Na]⁺(calcd for C₂₇H₂₇N₃O₄Na 480.1899)。 Low analytical spray mass spectrometry ( m/z , %): 480 [M+Na] ⁺ (100), 458 [M+1] ⁺ (82). High-resolution mass spectrum: m/z 480.1897 [M+Na] ⁺ (calcd for C ₂₇ H ₂₇ N ₃ O ₄ Na 480.1899).

【實施例三】N-(N-煙醯基-5-羥基-L-色氨醯基)-D-苯丙氨酸甲酯(N-(N-nicotinoyl-L-tryptophanyl)-D-phenylalanine methyl esters，化合物15a及15b)的製備方法如下： [Third Embodiment] N - (N - acyl-5-hydroxy -L- smoke color amino acyl) -D- phenylalanine methyl ester (N- (N-nicotinoyl-L -tryptophanyl) -D-phenylalanine The preparation methods of methyl esters , compounds 15a and 15b ) are as follows:

左旋色胺酸丙烯酸甲酯(L-tryptophan methyl ester)(1.0當量)與煙醯氯(nicotinoyl chloride)(1.1當量)進行反應，以吡啶(pyridine)作為溶媒，室溫下反應overnight的時間，經管柱層析進行純化〔溶媒系統MeOH/CHCl₃=1/20〕，得到第一個中間產物N-nicotinoyl-L-tryptophan methyl ester(產率：50.8%)。接著利用1M LiOH(2.0當量)進行皂化反應(saponification)，以四氫呋喃(THF)作溶媒，反應1小時後加入飽和碳酸氫鈉溶液，接著以乙酸乙酯(Ethyl acetate,EA)進行分配萃取，經萃取得到之水層加入1N鹽酸調整pH值至1~2，再做一次萃取動作，得到之EA層經減壓濃縮後即得第二個中間產物N-nicotinoyl-L-tryptophan(產率：93.5%)。最後再將第二個中間產物(1.0當量)與D-phenylalanine methyl ester(1.0當量)進行反應，以二氯甲烷(CH₂Cl₂)為溶媒，在苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)(2.0當量)及N,N-二異丙基乙胺(DIEA)(1.5當量)作用下在室溫下反應6小時，經以溶媒乙酸乙酯進行管柱層析純化，即可得到最終產物(化合物15a及15b)。 L-tryptophan methyl ester (1.0 eq.) was reacted with nicotinoyl chloride (1.1 eq.), using pyridine as a solvent, and reacted overnight at room temperature. Purification by column chromatography [solvent system MeOH/CHCl ₃ = 1/20] gave the first intermediate product N- nicotinoyl-L-tryptophan methyl ester (yield: 50.8%). Then, saponification was carried out using 1 M LiOH (2.0 eq.), and tetrahydrofuran (THF) was used as a solvent. After reacting for 1 hour, a saturated sodium hydrogencarbonate solution was added, followed by partition extraction with ethyl acetate (Ethyl acetate, EA). The extracted aqueous layer was added with 1N hydrochloric acid to adjust the pH to 1-2, and an extraction operation was carried out. The obtained EA layer was concentrated under reduced pressure to obtain a second intermediate product, N- nicotinoyl-L-tryptophan (yield: 93.5). %). Finally, a second intermediate (1.0 eq.) was reacted with D-phenylalanine methyl ester (1.0 eq.) with dichloromethane (CH ₂ Cl ₂ ) as the solvent in benzotriazole- N, N, N. ',N' -tetramethylurea hexafluorophosphate (HBTU) (2.0 eq.) and N,N -diisopropylethylamine (DIEA) (1.5 eq.) were reacted at room temperature for 6 hours. The solvent ethyl acetate was purified by column chromatography to obtain the final product ( compounds 15a and 15b ).

化合物15a：產率30%。白色粉狀固體，熔點：161-163℃。 Compound 15a: Yield 30%. White powdery solid, melting point: 161-163 ° C.

¹H NMR(C₅D₅N)：δ8.88(1H,s),8.66(1H,d,J=4.4Hz),8.21(1H,s,NH),7.98(1H,d,J=8.0Hz),7.72(1H,d,J=7.6Hz),7.35(1H,d,J=7.6Hz),7.31(1H,dd,J=7.8,2.0Hz),7.23-7.12(5H,m),7.08(1H,d,J=7.6Hz),6.85(1H,d,J=6.8Hz),6.35(1H,d,J=8.0Hz,NH),4.95(1H,ddd,J=14.4,7.6,7.6Hz),4.79(1H,dd,J=13.6,6.0Hz),3.63(3H,s),3.40(1H,dd,J=14.4,7.6Hz),3.23(1H,dd,J=14.4,7.6Hz),2.93(1H,dd,J=13.6,6.0Hz),2.82(1H,dd,J=13.6,6.0Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 8.88 (1H, s), 8.66 (1H, d, J = 4.4 Hz), 8.21. (1H, s, NH), 7.98 (1H, d, J = 8.0 Hz), 7.72 (1H, d, J = 7.6 Hz), 7.35 (1H, d, J = 7.6 Hz), 7.31 (1H, dd, J = 7.8, 2.0 Hz), 7.23 - 7.12 (5H, m), 7.08 (1H, d, J = 7.6 Hz), 6.85 (1H, d, J = 6.8 Hz), 6.35 (1H, d, J = 8.0 Hz, NH), 4.95 (1H, ddd, J = 14.4, 7.6, 7.6 Hz), 4.79 (1H, dd, J = 13.6, 6.0 Hz), 3.63 (3H, s), 3.40 (1H, dd, J = 14.4, 7.6 Hz), 3.23 (1H, dd, J = 14.4, 7.6 Hz), 2.93 (1H, dd, J = 13.6, 6.0 Hz), 2.82 (1H, dd, J = 13.6, 6.0 Hz).

¹³C NMR(C₅D₅N)：δ172.0(s),171.4(s),165.8(s),152.7(d),148.6(d),136.6(s),136.0(d),135.6(s),129.9(s),129.5(2×C,d),129.0(2×C,d),127.9(s),127.6(d),123.8(d),123.6(d),122.8(d),120.3(d),119.1(d),111.8(d),110.7(d),54.8(d),53.7(d),52.7(q),37.9(t),28.8(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.0 (s), 171.4 (s), 165.8 (s), 152.7 (d), 148.6 (d), 136.6 (s), 136.0 (d), 135.6 ( s), 129.9 (s), 129.5 (2 × C, d), 129.0 (2 × C, d), 127.9 (s), 127.6 (d), 123.8 (d), 123.6 (d), 122.8 (d) , 120.3 (d), 119.1 (d), 111.8 (d), 110.7 (d), 54.8 (d), 53.7 (d), 52.7 (q), 37.9 (t), 28.8 (t).

低解析噴灑式質譜(m/z，%)：471[M+1]⁺(100)。高解析度質譜：m/z 493.1850[M+Na]⁺(calcd for C₂₇H₂₆N₄O₄Na 493.1852)。 Low analytical spray mass spectrometry ( m/z , %): 471 [M+1] ⁺ (100). High-resolution mass spectrum: m/z 493.1850 [M+Na] ⁺ (calcd for C ₂₇ H ₂₆ N ₄ O ₄ Na 493.1852).

化合物15b：產率6%。白色粉狀固體，熔點：130-132℃。 Compound 15b: Yield 6%. White powdery solid, melting point: 130-132 ° C.

¹H NMR(C₅D₅N)：δ8.88(1H,s),8.69(1H,d,J=4.0Hz),8.25(1H,s,NH),7.99(1H,dd,J=6.0,2.0Hz),7.73(1H,d,J=8.0Hz),7.35-7.31(2H,m),7.20(1H,t,J=8.0Hz),7.13-7.06(6H,m),6.85(2H,d,J=6.8Hz),6.30(1H,d,J=7.6Hz,NH),4.92(1H,dd,J=14.6,7.6Hz),4.73(1H,dd,J=13.6,6.4Hz),3.65(3H,s),3.45(1H,dd,J=14.6,7.6Hz),3.21(1H,dd,J=14.6,7.6Hz),3.01(1H,dd,J=13.6,6.4Hz),2.92(1H,dd,J=13.6,6.4Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 8.88 (1H, s), 8.69 (1H, d, J = 4.0 Hz), 8.25 (1H, s, NH), 7.99 (1H, dd, J = 6.0 , 2.0 Hz), 7.73 (1H, d, J = 8.0 Hz), 7.35 - 7.31 (2H, m), 7.20 (1H, t, J = 8.0 Hz), 7.13 - 7.06 (6H, m), 6.85 (2H) , d, J = 6.8 Hz), 6.30 (1H, d, J = 7.6 Hz, NH), 4.92 (1H, dd, J = 14.6, 7.6 Hz), 4.73 (1H, dd, J = 13.6, 6.4 Hz) , 3.65 (3H, s), 3.45 (1H, dd, J = 14.6, 7.6 Hz), 3.21 (1H, dd, J = 14.6, 7.6 Hz), 3.01 (1H, dd, J = 13.6, 6.4 Hz), 2.92 (1H, dd, J = 13.6, 6.4 Hz).

¹³C NMR(C₅D₅N)：δ171.8(s),171.1(s),165.7(s),152.8(d),148.6(d),136.7(s),135.9(d),135.5(s),129.9(s),129.5(2×C,d),128.9(2×C,d),127.8(s),127.5(d),124.0(d),123.8(d),122.8(d),120.4(d),119.2(d),111.8(d),110.7(d),54.5(d),53.9(d),52.7(q),38.2(t),28.7(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 171.8 (s), 171.1 (s), 165.7 (s), 152.8 (d), 148.6 (d), 136.7 (s), 135.9 (d), 135.5 ( s), 129.9 (s), 129.5 (2 × C, d), 128.9 (2 × C, d), 127.8 (s), 127.5 (d), 124.0 (d), 123.8 (d), 122.8 (d) , 120.4 (d), 119.2 (d), 111.8 (d), 110.7 (d), 54.5 (d), 53.9 (d), 52.7 (q), 38.2 (t), 28.7 (t).

低解析噴灑式質譜(m/z，%)：471[M+1]⁺(100)。高解析度質譜：m/z 493.1849[M+Na]⁺(calcd for C₂₇H₂₆N₄O₄Na 493.1852)。 Low analytical spray mass spectrometry ( m/z , %): 471 [M+1] ⁺ (100). High resolution mass spectrum: m/z 493.1849 [M+Na] ⁺ (calcd for C ₂₇ H ₂₆ N ₄ O ₄ Na 493.1852).

【實施例四】N-(N-苄基-L-色氨醯基)-對-取代-D-苯丙氨酸甲酯(N-(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine methyl esters，化合物18a-23a與18b-23b)以及N-(N-苄基-L-色氨醯基)-3-環己基-D-丙氨酸甲酯(N-(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters，24a與24b)的製備方法如下： According to a fourth embodiment] [N - (N - benzyl-acyl -L- tryptophyl) - on - substituted -D- phenylalanine methyl ester (N - (N -benzoyl-L -tryptophanyl) - para -substituted- D-phenylalanine methyl esters, compounds 18a-23a and 18b-23b) and N- ( N -benzyl-L-chromomethylamino)-3-cyclohexyl-D-alanine methyl ester ( N -( N - The preparation methods of benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters, 24a and 24b) are as follows:

二碳酸二叔丁酯-D-苯丙氨酸(Boc-D-phenyl alanine)衍生物(1.0當量)加入20%三氟乙酸(TFA)(TFA/CH₂Cl₂=1/4)切除Boc保護基團，室溫下反應30分鐘，待反應結束後經減壓濃縮除去TFA及溶媒，得到D-苯丙氨酸衍生物。D-苯丙氨酸衍生物或(2R)-2-氨基-3-環己基丙酸(3-Cyclohexyl-D-alanine)加入甲醇(MeOH)做為溶媒，並加入濃硫酸(conc.H₂SO₄)1mL，加熱迴流2小時，進行酯化反應(esterification)。反應結束後加入預先冰浴的逆滲透(RO)水10mL，隨後加入氨水做pH值調整動作，利用pH試紙測定觀察，將pH值調至7-9，接著以氯仿(CHCl₃)進行分配萃取，取得之氯仿(CHCl₃)層經減壓濃縮後即可得到中間產物丙烯酸甲酯D-***酸丙烯酸甲酯(D-phenylalanine methyl ester)衍生物。D-苯丙氨酸甲酯鹽酸鹽(D-phenylalanine methyl ester)衍生物(1.0當量)與N-苯甲醯-左旋色胺酸(N-benzoyl-L-tryptophan)(1.0當量)衍生物進行反應，以二氯甲烷(CH₂Cl₂)為溶媒，隨後加入苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)(2.0當量)及N,N-二異丙基乙胺(DIEA)(1.5當量)在室溫下反應6小時，經以溶媒系統正已烷/乙酸乙酯(6：4)分別進行管柱層析純化，即可得到最終產物(化合物18a-24a與18b-24b)。 Bic-D-phenyl alanine derivative (1.0 eq.) was added to 20% trifluoroacetic acid (TFA) (TFA/CH ₂ Cl ₂ = 1/4) to remove Boc. The protective group was reacted at room temperature for 30 minutes, and after completion of the reaction, the TFA and the solvent were removed by concentration under reduced pressure to give a D-phenylalanine derivative. D-phenylalanine derivative or (2R)-2-amino-3-cyclohexylpropanoic acid (3-Cyclohexyl-D-alanine) is added to methanol (MeOH) as a solvent, and concentrated sulfuric acid (conc.H _{2 is} added) SO ₄ ) 1 mL, heated under reflux for 2 hours, and subjected to esterification. After the reaction, 10 mL of reverse osmosis (RO) water in a pre-ice bath was added, followed by the addition of ammonia water for pH adjustment, and the pH was adjusted to 7-9 by pH test paper, followed by partition extraction with chloroform (CHCl ₃ ). The obtained chloroform (CHCl ₃ ) layer was concentrated under reduced pressure to obtain an intermediate product of methyl acrylate D-phenylalanine methyl ester. D- phenylalanine methyl ester hydrochloride (D-phenylalanine methyl ester) derivative (1.0 eq.) And N - benzoyl - L-tryptophan (N -benzoyl-L-tryptophan) (1.0 eq.) Derivative The reaction was carried out with dichloromethane (CH ₂ Cl ₂ ) as a solvent, followed by the addition of benzotriazole- N,N,N',N' -tetramethyluronium hexafluorophosphate (HBTU) (2.0 equivalents) and N,N -diisopropylethylamine (DIEA) (1.5 eq.) was reacted at room temperature for 6 hours, and purified by column chromatography using solvent system n-hexane/ethyl acetate (6:4). The final product ( compounds 18a-24a and 18b-24b ) was obtained.

N-(N-苄基-L-色氨醯基)-對-硝基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-nitro-D-phenylalanine methyl esters(化合物18a和18b) N - (N - benzyl-acyl -L- tryptophyl) - on - nitro -D- phenylalanine methyl ester (N- (N-Benzoyl-L -tryptophanyl) -para-nitro-D-phenylalanine methyl Esters (compounds 18a and 18b)

化合物18a：產率42%。白色粉狀固體，熔點：240-242℃。 Compound 18a: Yield 42%. White powdery solid, melting point: 240-242 ° C.

¹H NMR(C₅D₅N)：δ11.91(1H,s,NH),9.76(1H,d,J=8.0Hz,NH),9.24(1H,d,J=8.0Hz,NH),8.09(2H,d,J=8.0Hz),7.98(2H,d,J=8.0Hz),7.90(1H,d,J=8.0Hz),7.56(1H,d,J=8.0Hz),7.45(1H,s),7.36(1H,t,J=8.0Hz),7.30-7.24(3H,m),7.17-7.14(3H,m),5.66(1H,dd,J=14.2,7.2Hz),5.31(1H,dd,J=13.8,7.2Hz),3.80(1H,dd,J=14.2,7.2Hz),3.62(1H,dd, J=14.2,7.2Hz),3.55(3H,s),3.22(1H,dd,J=13.8,7.2Hz),3.15(1H,dd,J=13.8,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.91 (1H, s, NH), 9.76 (1H, d, J = 8.0 Hz, NH), 9.24 (1H, d, J = 8.0 Hz, NH), 8.09 (2H, d, J = 8.0 Hz), 7.98 (2H, d, J = 8.0 Hz), 7.90 (1H, d, J = 8.0 Hz), 7.56 (1H, d, J = 8.0 Hz), 7.45 ( 1H, s), 7.36 (1H, t, J = 8.0 Hz), 7.30-7.24 (3H, m), 7.17-7.14 (3H, m), 5.66 (1H, dd, J = 14.2, 7.2 Hz), 5.31 (1H, dd, J = 13.8, 7.2 Hz), 3.80 (1H, dd, J = 14.2, 7.2 Hz), 3.62 (1H, dd, J = 14.2, 7.2 Hz), 3.55 (3H, s), 3.22 ( 1H, dd, J = 13.8, 7.2 Hz), 3.15 (1H, dd, J = 13.8, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),171.7(s),167.9(s),147.1(s),145.4(s),137.5(s),135.4(s),131.5(d),130.7(2×C,d),128.6(s),128.6(2×C,d),128.1(2×C,d),124.5(d),123.7(2×C,d),121.9(d),119.4(d),119.3(d),112.1(d),111.2(s),55.5(d),53.8(d),52.1(q),37.5(t),28.9(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.8 (s), 171.7 (s), 167.9 (s), 147.1 (s), 145.4 (s), 137.5 (s), 135.4 (s), 131.5 ( d), 130.7 (2 × C, d), 128.6 (s), 128.6 (2 × C, d), 128.1 (2 × C, d), 124.5 (d), 123.7 (2 × C, d), 121.9 (d), 119.4(d), 119.3(d), 112.1(d), 111.2(s), 55.5(d), 53.8(d), 52.1(q), 37.5(t), 28.9(t).

低解析噴灑式質譜(m/z，%)：537[M+Na]⁺(100)。高解析度質譜：m/z 537.1754[M+Na]⁺(calcd for C₂₈H₂₆N₄O₆Na 537.1750)。 Low analytical spray mass spectrometry ( m/z , %): 537 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 537.1754 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ N ₄ O ₆ Na 537.1750).

化合物18b：產率8%。白色粉狀固體，熔點：182-184℃。 Compound 18b: Yield 8%. White powdery solid, melting point: 182-184 °C.

¹H NMR(C₅D₅N)：δ11.81(1H,s,NH),9.72(1H,d,J=8.0Hz,NH),9.29(1H,d,J=8.0Hz,NH),8.10(2H,d,J=8.0Hz),7.97(2H,d,J=8.0Hz),7.74(1H,d,J=8.0Hz),7.49(1H,d,J=8.0Hz),7.44(1H,s),7.41-7.37(3H,m),7.32(2H,t,J=7.6Hz),7.20(1H,t,J=8.0Hz),7.05(1H,t,J=8.0Hz),5.65(1H,dd,J=14.8,7.2Hz),5.31(1H,dd,J=13.6,8.0Hz),3.77(1H,dd,J=14.8,7.2Hz),3.62(3H,s),3.61(1H,dd,J=14.8,7.2Hz),3.39(1H,dd,J=13.6,8.0Hz),3.21(1H,dd,J=13.6,8.0Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.81 (1H, s, NH), 9.72 (1H, d, J = 8.0 Hz, NH), 9.29 (1H, d, J = 8.0 Hz, NH), 8.10 (2H, d, J = 8.0 Hz), 7.97 (2H, d, J = 8.0 Hz), 7.74 (1H, d, J = 8.0 Hz), 7.49 (1H, d, J = 8.0 Hz), 7.44 ( 1H, s), 7.41-7.37 (3H, m), 7.32 (2H, t, J = 7.6 Hz), 7.20 (1H, t, J = 8.0 Hz), 7.05 (1H, t, J = 8.0 Hz), 5.65 (1H, dd, J = 14.8, 7.2 Hz), 5.31 (1H, dd, J = 13.6, 8.0 Hz), 3.77 (1H, dd, J = 14.8, 7.2 Hz), 3.62 (3H, s), 3.61 (1H, dd, J = 14.8, 7.2 Hz), 3.39 (1H, dd, J = 13.6, 8.0 Hz), 3.21 (1H, dd, J = 13.6, 8.0 Hz).

¹³C NMR(C₅D₅N)：δ173.0(s),171.9(s),168.0(s),147.1(s),145.4(s),137.5(s),135.4(s),131.6(d),130.9(2×C,d),128.6(2×C,d),128.5(s),128.1(2×C,d),124.4(d),123.7(2×C,d),121.7(d),119.2(d),119.1(d),111.9(d),111.1(s),55.3(d),53.7(d),52.2(q),37.8(t),28.5(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 173.0 (s), 171.9 (s), 168.0 (s), 147.1 (s), 145.4 (s), 137.5 (s), 135.4 (s), 131.6 ( d), 130.9 (2 × C, d), 128.6 (2 × C, d), 128.5 (s), 128.1 (2 × C, d), 124.4 (d), 123.7 (2 × C, d), 121.7 (d), 119.2(d), 119.1(d), 111.9(d), 111.1(s), 55.3(d), 53.7(d), 52.2(q), 37.8(t), 28.5(t).

低解析噴灑式質譜(m/z，%)：537[M+Na]⁺(100)。高解析度質譜：m/z 537.1753[M+Na]⁺(calcd for C₂₈H₂₆N₄O₆Na 537.1750)。 Low analytical spray mass spectrometry ( m/z , %): 537 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 537.1753 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ N ₄ O ₆ Na 537.1750).

N-(N-苄基-L-色氨醯基)-對-甲基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-methyl-D-phenylalanine methyl esters ，化合物19a和19b) N - (N - benzyl-acyl -L- tryptophyl) - on - -D- phenylalanine methyl ester (N- (N-Benzoyl-L -tryptophanyl) -para-methyl-D-phenylalanine methyl Esters , compounds 19a and 19b)

化合物19a：產率50%。白色粉狀固體，熔點：185-187℃。圓二色光譜(c 0.18mM,MeOH)nm(mdeg)：232(-1.30),224(-0.28)nm。 Compound 19a: Yield 50%. White powdery solid, melting point: 185-187 °C. Circular dichroism ( c 0.18 mM, MeOH) nm (mdeg): 232 (-1.30), 224 (-0.28) nm.

¹H NMR(C₅D₅N)：δ11.82(1H,s,NH),9.35(1H,d,J=8.0 Hz,NH),9.07(1H,d,J=8.0Hz,NH),8.04(2H,d,J=8.0Hz),7.87(2H,d,J=8.0Hz),7.53(1H,d,J=7.6Hz),7.36-7.34(2H,m),7.28-7.20(3H,m),7.14(1H,t,J=7.6Hz),7.08(1H,d,J=7.6Hz),6.99(2H,d,J=7.6Hz),5.68(1H,dd,J=14.4,6.8Hz),5.22(1H,dd,J=13.8,7.2Hz),3.80(1H,dd,J=14.4,6.8Hz),3.64(1H,dd,J=14.4,6.8Hz),3.52(3H,s),3.16(1H,dd,J=13.8,7.2Hz),3.07(1H,dd,J=13.8,7.2Hz),2.11(3H,s)。 ¹ H NMR (C ₅ D ₅ N): δ 11.82 (1H, s, NH), 9.35 (1H, d, J = 8.0 Hz, NH), 9.07 (1H, d, J = 8.0 Hz, NH), 8.04 (2H, d, J = 8.0 Hz), 7.87 (2H, d, J = 8.0 Hz), 7.53 (1H, d, J = 7.6 Hz), 7.36-7.34 (2H, m), 7.28-7.20 (3H , m), 7.14 (1H, t, J = 7.6 Hz), 7.08 (1H, d, J = 7.6 Hz), 6.99 (2H, d, J = 7.6 Hz), 5.68 (1H, dd, J = 14.4, 6.8 Hz), 5.22 (1H, dd, J = 13.8, 7.2 Hz), 3.80 (1H, dd, J = 14.4, 6.8 Hz), 3.64 (1H, dd, J = 14.4, 6.8 Hz), 3.52 (3H, s), 3.16 (1H, dd, J = 13.8, 7.2 Hz), 3.07 (1H, dd, J = 13.8, 7.2 Hz), 2.11 (3H, s).

¹³C NMR(C₅D₅N)：δ172.7(s),172.4(s),167.8(s),137.5(s),135.4(s),134.3(s),131.4(d),129.7(2×C,d),129.5(2×C,d),128.7(s),128.5(2×C,d),128.0(2×C,d),124.4(d),121.8(d),119.3(2×C,d),112.0(d),111.2(s),55.3(d),54.5(d),51.9(q),37.6(t),29.0(t),20.9(q)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.7 (s), 172.4 (s), 167.8 (s), 137.5 (s), 135.4 (s), 134.3 (s), 131.4 (d), 129.7 ( 2 × C, d), 129.5 (2 × C, d), 128.7 (s), 128.5 (2 × C, d), 128.0 (2 × C, d), 124.4 (d), 121.8 (d), 119.3 (2 x C, d), 112.0 (d), 111.2 (s), 55.3 (d), 54.5 (d), 51.9 (q), 37.6 (t), 29.0 (t), 20.9 (q).

低解析噴灑式質譜(m/z，%)：506[M+Na]⁺(100)。高解析度質譜：m/z 506.2059[M+Na]⁺(calcd for C₂₉H₂₉N₃O₄Na 506.2056)。 Low analytical spray mass spectrometry ( m/z , %): 506 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 506.2059 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₄ Na 506.2056).

化合物19b：產率16%。白色粉狀固體，熔點：197-199℃。圓二色光譜(c 0.18mM,MeOH)nm(mdeg)：233(1.75),221(-1.81),213(-0.42)nm。 Compound 19b: Yield 16%. White powdery solid, melting point: 197-199 ° C. Circular dichroism ( c 0.18 mM, MeOH) nm (mdeg): 233 (1.75), 221 (-1.81), 213 (-0.42) nm.

¹H NMR(C₅D₅N)：δ11.79(1H,s,NH),9.52(1H,d,J=8.0Hz,NH),9.16(1H,d,J=8.0Hz,NH),8.09(2H,d,J=8.0Hz),7.78(2H,d,J=8.0Hz),7.48(1H,d,J=8.0Hz),7.46(1H,d,J=1.6Hz),7.38(1H,t,J=8.0Hz),7.30(2H,t,J=8.0Hz),7.22-7.15(3H,m),7.07(1H,d,J=8.0Hz),6.92(2H,d,J=7.6Hz),5.70(1H,dd,J=14.4,7.2Hz),5.24(1H,dd,J=13.8,7.2Hz),3.80(1H,dd,J=14.4,7.2Hz),3.65(1H,dd,J=14.4,7.2Hz),3.58(3H,s),3.27(1H,dd,J=13.8,7.2Hz),3.14(1H,dd,J=13.8,7.2Hz),2.04(3H,s)。 ¹ H NMR (C ₅ D ₅ N): δ 11.79 (1H, s, NH), 9.52 (1H, d, J = 8.0 Hz, NH), 9.16 (1H, d, J = 8.0 Hz, NH), 8.09 (2H, d, J = 8.0 Hz), 7.78 (2H, d, J = 8.0 Hz), 7.48 (1H, d, J = 8.0 Hz), 7.46 (1H, d, J = 1.6 Hz), 7.38 ( 1H, t, J = 8.0 Hz), 7.30 (2H, t, J = 8.0 Hz), 7.22 - 7.15 (3H, m), 7.07 (1H, d, J = 8.0 Hz), 6.92 (2H, d, J = 7.6 Hz), 5.70 (1H, dd, J = 14.4, 7.2 Hz), 5.24 (1H, dd, J = 13.8, 7.2 Hz), 3.80 (1H, dd, J = 14.4, 7.2 Hz), 3.65 (1H) , dd, J = 14.4, 7.2 Hz), 3.58 (3H, s), 3.27 (1H, dd, J = 13.8, 7.2 Hz), 3.14 (1H, dd, J = 13.8, 7.2 Hz), 2.04 (3H, s).

¹³C NMR(C₅D₅N)：δ172.7(s),172.5(s),167.9(s),137.5(s),136.2(s),135.3(s),134.3(s),131.4(d),129.7(2×C,d),129.4(2×C,d),128.7(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.7(d),119.2(2×C,d),111.9(d),111.2(s),55.1(d),54.6(d),51.9(q),37.8(t),28.6(t),20.9(q)。 ¹³ C NMR (C ₅ D ₅ N): δ 172.7 (s), 172.5 (s), 167.9 (s), 137.5 (s), 136.2 (s), 135.3 (s), 134.3 (s), 131.4 ( d), 129.7 (2 × C, d), 129.4 (2 × C, d), 128.7 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.7 (d), 119.2 (2 × C, d), 111.9 (d), 111.2 (s), 55.1 (d), 54.6 (d), 51.9 (q), 37.8 (t), 28.6 (t), 20.9 ( q).

低解析噴灑式質譜(m/z，%)：506[M+Na]⁺(100)。高解析度質譜：m/z 506.2060[M+Na]⁺(calcd for C₂₉H₂₉N₃O₄Na 506.2056)。 Low analytical spray mass spectrometry ( m/z , %): 506 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 506.2060 [M+Na] ⁺ (calcd for C ₂₉ H ₂₉ N ₃ O ₄ Na 506.2056).

N-(N-苄基-L-色氨醯基)-對-氟基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-fluoro-D-phenylalanine methyl esters ，化合物20a和20b) N - (N - benzyl-acyl -L- tryptophyl) - on - -D- fluoro-phenylalanine methyl ester (N- (N-Benzoyl-L -tryptophanyl) -para-fluoro-D-phenylalanine methyl Esters , compounds 20a and 20b)

化合物20a：產率45%。白色粉狀固體，熔點：178-180℃。圖二色光譜(c 0.12mM,MeOH)nm(mdeg)：232(-0.76),223(0.68)nm。 Compound 20a: Yield 45%. White powdery solid, melting point: 178-180 ° C. Figure 2 color spectrum ( c 0.12 mM, MeOH) nm (mdeg): 232 (-0.76), 223 (0.68) nm.

¹H NMR(C₅D₅N)：δ11.84(1H,s,NH),9.40(1H,d,J=7.6Hz,NH),9.15(1H,d,J=7.6Hz,NH),8.07(2H,d,J=7.6Hz),7.90(2H,d,J=7.6Hz),7.54(1H,d,J=7.6Hz),7.40(1H,s),7.35(1H,t,J=7.2Hz),7.29-7.25(3H,m),7.25-7.20(3H,m),7.15(1H,t,J=7.6Hz),7.08(1H,d,J=6.4Hz),6.89(2H,t,J=8.0Hz),5.68(1H,dd,J=14.2,6.8Hz),5.19(1H,dd,J=13.6,6.8Hz),3.79(1H,dd,J=14.2,6.8Hz),3.65(1H,dd,J=14.2,6.8Hz),3.58(3H,s),3.27(1H,dd,J=13.8,6.8Hz),3.13(1H,dd,J=13.6,6.8Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.84 (1H, s, NH), 9.40 (1H, d, J = 7.6 Hz, NH), 9.15 (1H, d, J = 7.6 Hz, NH), 8.07 (2H, d, J = 7.6 Hz), 7.90 (2H, d, J = 7.6 Hz), 7.54 (1H, d, J = 7.6 Hz), 7.40 (1H, s), 7.35 (1H, t, J = 7.2 Hz), 7.29-7.25 (3H, m), 7.25-7.20 (3H, m), 7.15 (1H, t, J = 7.6 Hz), 7.08 (1H, d, J = 6.4 Hz), 6.89 (2H) , t, J = 8.0 Hz), 5.68 (1H, dd, J = 14.2, 6.8 Hz), 5.19 (1H, dd, J = 13.6, 6.8 Hz), 3.79 (1H, dd, J = 14.2, 6.8 Hz) , 3.65 (1H, dd, J = 14.2, 6.8 Hz), 3.58 (3H, s), 3.27 (1H, dd, J = 13.8, 6.8 Hz), 3.13 (1H, dd, J = 13.6, 6.8 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),172.1(s),167.9(s),162.1(s,J _C-F=242.2Hz),137.5(s),135.4(s),133.4(s,J _C-F=3.0Hz),131.4(d),128.7(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.9(d),119.3(2×C,d),115.4(2×C,d,J _C-F=21.4Hz),112.0(d),111.2(s),55.4(d),54.4(d),51.9(q),37.1(t),29.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.7 (s), 172.1 (s), 167.9 (s), 162.1 (s, J CF = 242.2Hz), 137.5 (s), 135.4 (s), 133.4 (s, J _CF = 3.0 Hz), 131.4 (d), 128.7 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.9 (d), 119.3 ( 2 × C, d), 115.4 (2 × C, d, J _CF = 21.4 Hz), 112.0 (d), 111.2 (s), 55.4 (d), 54.4 (d), 51.9 (q), 37.1 (t ), 29.0(t).

低解析噴灑式質譜(m/z，%)：510[M+Na]⁺(100)。高解析度質譜：m/z 510.1808[M+Na]⁺(calcd for C₂₈H₂₆FN₃O₄Na 510.1805)。 Low analytical spray mass spectrometry ( m/z , %): 510 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 510.1808 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ FN ₃ O ₄ Na 510.1805).

化合物20b：產率14%。白色粉狀固體，熔點：184-186℃。圓二色光譜(c 0.12mM,MeOH)nm(mdeg)：233(0.69),221(-0.55)nm。 Compound 20b: Yield 14%. White powdery solid, melting point: 184-186 ° C. Circular dichroism ( c 0.12 mM, MeOH) nm (mdeg): 233 (0.69), 221 (-0.55) nm.

¹H NMR(C₅D₅N)：δ11.80(1H,s,NH),9.60(1H,d,J=8.0Hz,NH),9.22(1H,d,J=8.0Hz,NH),8.11(2H,d,J=7.6Hz),7.77(2H,d,J=7.6Hz),7.49(1H,d,J=7.6Hz),7.46(1H,s),7.37(1H,t,J=7.6Hz),7.30(1H,t,J=7.6Hz),7.25-7.20(3H,m),7.06(1H,t,J=7.6Hz),6.89(2H,t,J=8.0Hz),5.68(1H,dd,J=14.6,7.2Hz),5.22(1H,dd,J=13.8,7.2Hz),3.79(1H,dd,J=14.6,7.2Hz),3.65(1H,dd,J=14.6,7.2Hz),3.58(3H,s),3.27(1H,dd,J=13.8, 7.2Hz),3.13(1H,dd,J=13.8,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.80 (1H, s, NH), 9.60 (1H, d, J = 8.0 Hz, NH), 9.22 (1H, d, J = 8.0 Hz, NH), 8.11 (2H, d, J = 7.6 Hz), 7.77 (2H, d, J = 7.6 Hz), 7.49 (1H, d, J = 7.6 Hz), 7.46 (1H, s), 7.37 (1H, t, J = 7.6 Hz), 7.30 (1H, t, J = 7.6 Hz), 7.25-7.20 (3H, m), 7.06 (1H, t, J = 7.6 Hz), 6.89 (2H, t, J = 8.0 Hz), 5.68 (1H, dd, J = 14.6, 7.2 Hz), 5.22 (1H, dd, J = 13.8, 7.2 Hz), 3.79 (1H, dd, J = 14.6, 7.2 Hz), 3.65 (1H, dd, J = 14.6, 7.2 Hz), 3.58 (3H, s), 3.27 (1H, dd, J = 13.8, 7.2 Hz), 3.13 (1H, dd, J = 13.8, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),172.3(s),167.9(s),162.1(s,J _C-F=242.0Hz),137.5(s),135.5(s),133.4(s,J _C-F=3.2Hz),131.6(d,J _C-F=8.0Hz),131.5(d),128.6(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.7(d),119.2(d),119.1(d),115.4(2×C,d,J _C-F=21.0Hz),111.9(d),111.2(s),55.2(d),54.4(d),52.0(q),37.3(t),28.6(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.8 (s), 172.3 (s), 167.9 (s), 162.1 (s, J CF = 242.0Hz), 137.5 (s), 135.5 (s), 133.4 (s, J _CF = 3.2 Hz), 131.6 (d, J _CF = 8.0 Hz), 131.5 (d), 128.6 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.7 (d) , 119.2 (d), 119.1 (d), 115.4 (2 × C, d, J CF = 21.0Hz), 111.9 (d), 111.2 (s), 55.2 (d), 54.4 (d), 52.0 (q), 37.3 (t), 28.6 (t).

低解析噴灑式質譜(m/z，%)：510[M+Na]⁺(100)。高解析度質譜：m/z 510.1809[M+Na]⁺(calcd for C₂₈H₂₆FN₃O₄Na 510.1805)。 Low analytical spray mass spectrometry ( m/z , %): 510 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 510.1809 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ FN ₃ O ₄ Na 510.1805).

N-(N-苄基-L-色氨醯基)-對-氯基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-chloro-D-phenylalanine methyl esters ，化合物21a和21b) N - (N - benzyl-acyl -L- tryptophyl) - on - chloro - -D- phenylalanine methyl ester (N- (N-Benzoyl-L -tryptophanyl) -para-chloro-D-phenylalanine methyl Esters , compounds 21a and 21b)

化合物21a：產率37%。白色粉狀固體，熔點：190-191℃。 Compound 21a: Yield 37%. White powdery solid, melting point: 190-191 °C.

¹H NMR(C₅D₅N)：δ11.86(1H,s,NH),9.42(1H,d,J=8.0Hz,NH),9.16(1H,d,J=8.0Hz,NH),8.07(2H,d,J=8.0Hz),7.90(1H,d,J=8.0Hz),7.54(1H,d,J=7.6Hz),7.40(1H,d,J=1.6Hz),7.35(1H,t,J=7.2Hz),7.29-7.23(3H,m),7.19-7.14(3H,m),7.02(2H,d,J=8.0Hz),5.68(1H,dd,J=14.2,7.2Hz),5.18(1H,dd,J=14.0,7.2Hz),3.79(1H,dd,J=14.2,7.2Hz),3.64(1H,dd,J=14.2,7.2Hz),3.51(3H,s),3.11(1H,dd,J=14.0,7.2Hz),3.03(1H,dd,J=14.0,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.86 (1H, s, NH), 9.42 (1H, d, J = 8.0 Hz, NH), 9.16 (1H, d, J = 8.0 Hz, NH), 8.07 (2H, d, J = 8.0 Hz), 7.90 (1H, d, J = 8.0 Hz), 7.54 (1H, d, J = 7.6 Hz), 7.40 (1H, d, J = 1.6 Hz), 7.35 ( 1H, t, J = 7.2 Hz), 7.29-7.23 (3H, m), 7.19-7.14 (3H, m), 7.02 (2H, d, J = 8.0 Hz), 5.68 (1H, dd, J = 14.2, 7.2 Hz), 5.18 (1H, dd, J = =14.0, 7.2 Hz), 3.79 (1H, dd, J = 14.2, 7.2 Hz), 3.64 (1H, dd, J = 14.2, 7.2 Hz), 3.51 (3H, s), 3.11 (1H, dd, J = 14.0, 7.2 Hz), 3.03 (1H, dd, J = 14.0, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),172.0(s),167.9(s),137.5(s),136.2(s),135.5(s),132.6(s),131.5(2×C,d),131.4(2×C,d),128.8(2×C,d),128.7(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.9(d),119.3(2×C,d),112.0(d),111.2(s),55.4(d),54.2(d),51.9(q),37.2(t),29.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.7 (s), 172.0 (s), 167.9 (s), 137.5 (s), 136.2 (s), 135.5 (s), 132.6 (s), 131.5 ( 2 × C, d), 131.4 (2 × C, d), 128.8 (2 × C, d), 128.7 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 124.4 ( d), 121.9(d), 119.3 (2×C, d), 112.0(d), 111.2(s), 55.4(d), 54.2(d), 51.9(q), 37.2(t), 29.0(t ).

低解析噴灑式質譜(m/z，%)：526[M+Na]⁺(100)。高解析度質譜：m/z 526.1506[M+Na]⁺(calcd for C₂₈H₂₆ClN₃O₄Na 526.1509)。 Low analytical spray mass spectrometry ( m/z , %): 526 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 526.1506 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ ClN ₃ O ₄ Na 526.1509).

化合物21b：產率9%。白色粉狀固體，熔點：174-176℃。 Compound 21b: Yield 9%. White powdery solid, melting point: 174-176 °C.

¹H NMR(C₅D₅N)：δ11.80(1H,s,NH),9.62(1H,d,J=8.0Hz,NH),9.23(1H,d,J=8.0Hz,NH),8.11(2H,d,J=8.0Hz),7.77(1H,d,J= 8.0Hz),7.49(1H,d,J=8.0Hz),7.46(1H,br.s),7.38(1H,t,J=8.0Hz),7.31(2H,t,J=8.0Hz),7.22-7.19(3H,m),7.14(2H,t,J=8.0Hz),7.06(1H,d,J=8.0Hz),5.68(1H,dd,J=14.6,7.2Hz),5.22(1H,dd,J=13.8,7.6Hz),3.79(1H,dd,J=14.6,7.2Hz),3.64(1H,dd,J=14.6,7.2Hz),3.58(3H,s),3.26(1H,dd,J=13.8,7.6Hz),3.12(1H,dd,J=13.8,7.6Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.80 (1H, s, NH), 9.62 (1H, d, J = 8.0 Hz, NH), 9.23 (1H, d, J = 8.0 Hz, NH), 8.11 (2H, d, J = 8.0 Hz), 7.77 (1H, d, J = 8.0 Hz), 7.49 (1H, d, J = 8.0 Hz), 7.46 (1H, br.s), 7.38 (1H, t , J = 8.0 Hz), 7.31 (2H, t, J = 8.0 Hz), 7.22-7.19 (3H, m), 7.14 (2H, t, J = 8.0 Hz), 7.06 (1H, d, J = 8.0 Hz) ), 5.68 (1H, dd, J = 14.6, 7.2 Hz), 5.22 (1H, dd, J = 13.8, 7.6 Hz), 3.79 (1H, dd, J = 14.6, 7.2 Hz), 3.64 (1H, dd, J = 14.6, 7.2 Hz), 3.58 (3H, s), 3.26 (1H, dd, J = 13.8, 7.6 Hz), 3.12 (1H, dd, J = 13.8, 7.6 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),172.2(s),168.0(s),137.5(s),136.3(s),135.5(s),132.5(s),131.5(d),131.4(2×C,d),128.7(s),128.7(2×C,d),128.6(2×C,d),124.4(d),121.7(d),119.2(d),119.1(d),111.9(d),111.2(s),55.2(d),54.2(d),52.0(q),37.4(t),28.6(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.8 (s), 172.2 (s), 168.0 (s), 137.5 (s), 136.3 (s), 135.5 (s), 132.5 (s), 131.5 ( d), 131.4 (2 × C, d), 128.7 (s), 128.7 (2 × C, d), 128.6 (2 × C, d), 124.4 (d), 121.7 (d), 119.2 (d), 119.1(d), 111.9(d), 111.2(s), 55.2(d), 54.2(d), 52.0(q), 37.4(t), 28.6(t).

N-(N-苄基-L-色氨醯基)-對-溴基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-bromo-D-phenylalanine methyl esters ，化合物22a和22b) N- ( N -benzyl -L-tryptophanyl)-p-bromo-D-phenylalanine methyl ( N-(N-Benzoyl-L-tryptophanyl)-para-bromo-D-phenylalanine methyl Esters , compounds 22a and 22b)

化合物22a：產率23%。白色粉狀固體，熔點：190-191℃。圓二色光譜(c 0.15mM,MeOH)nm(mdeg)：232(-0.41),223(0.15)nm。 Compound 22a: Yield 23%. White powdery solid, melting point: 190-191 °C. Circular dichroism ( c 0.15 mM, MeOH) nm (mdeg): 232 (-0.41), 223 (0.15) nm.

¹H NMR(C₅D₅N)：δ11.85(1H,s,NH),9.40(1H,d,J=8.0Hz,NH),9.12(1H,d,J=8.0Hz,NH),8.07(2H,d,J=7.2Hz),7.90(1H,d,J=7.6Hz),7.54(1H,d,J=7.6Hz),7.41(1H,br.s),7.38-7.32(3H,m),7.28(1H,t,J=7.2Hz),7.24(1H,t,J=7.6Hz),7.15(1H,t,J=8.0Hz),6.97(2H,d,J=8.0Hz),5.67(1H,dd,J=14.4,7.2Hz),5.17(1H,dd,J=13.8,7.2Hz),3.79(1H,dd,J=14.4,7.2Hz),3.64(1H,dd,J=14.4,7.2Hz),3.51(3H,s),3.09(1H,dd,J=13.8,7.2Hz),3.02(1H,dd,J=13.8,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.85 (1H, s, NH), 9.40 (1H, d, J = 8.0 Hz, NH), 9.12 (1H, d, J = 8.0 Hz, NH), 8.07 (2H, d, J = 7.2 Hz), 7.90 (1H, d, J = 7.6 Hz), 7.54 (1H, d, J = 7.6 Hz), 7.41 (1H, br.s), 7.38-7.32 (3H , m), 7.28 (1H, t, J = 7.2 Hz), 7.24 (1H, t, J = 7.6 Hz), 7.15 (1H, t, J = 8.0 Hz), 6.97 (2H, d, J = 8.0 Hz) ), 5.67 (1H, dd, J = 14.4, 7.2 Hz), 5.17 (1H, dd, J = 13.8, 7.2 Hz), 3.79 (1H, dd, J = 14.4, 7.2 Hz), 3.64 (1H, dd, J = 14.4, 7.2 Hz), 3.51 (3H, s), 3.09 (1H, dd, J = 13.8, 7.2 Hz), 3.02 (1H, dd, J = 13.8, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),172.0(s),167.8(s),137.5(s),136.7(s),135.4(s),131.8(2×C,d),131.7(2×C,d),131.5(d),128.7(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.9(d),120.9(s),119.3(2×C,d),112.0(d),111.2(s),55.4(d),54.1(d),51.9(q),37.2(t),29.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.7 (s), 172.0 (s), 167.8 (s), 137.5 (s), 136.7 (s), 135.4 (s), 131.8 (2 × C, d ), 131.7 (2 × C, d), 131.5 (d), 128.7 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.9 (d), 120.9 (s), 119.3 (2 x C, d), 112.0 (d), 111.2 (s), 55.4 (d), 54.1 (d), 51.9 (q), 37.2 (t), 29.0 (t).

低解析噴灑式質譜(m/z，%)：570[M+Na]⁺(100),572 [M+2+Na]⁺(100)。高解析度質譜：m/z 570.1008[M+Na]⁺(calcd for C₂₈H₂₆BrN₃O₄Na 570.1004)。 Low analytical spray mass spectrometry ( m/z , %): 570 [M+Na] ⁺ (100), 572 [M+2+Na] ⁺ (100). High Resolution Mass Spectrum: m / z 570.1008 [M + Na] + (calcd for C 28 H 26 BrN 3 O 4 Na 570.1004).

化合物22b：產率8%。白色粉狀固體，熔點：174-176℃。圓二色光譜(c 0.18mM,MeOH)nm(mdeg)：241(0.49),232(0.24),220(-0.75),214(-0.28)nm。 Compound 22b: Yield 8%. White powdery solid, melting point: 174-176 °C. Circular dichroism ( c 0.18 mM, MeOH) nm (mdeg): 241 (0.49), 232 (0.24), 220 (-0.75), 214 (-0.28) nm.

¹H NMR(C₅D₅N)：δ11.81(1H,s,NH),9.62(1H,d,J=8.0Hz,NH),9.23(1H,d,J=8.0Hz,NH),8.11(2H,d,J=8.0Hz),7.76(1H,d,J=7.6Hz),7.49(1H,d,J=7.6Hz),7.45(1H,d,J=1.6Hz),7.39(1H,t,J=7.6Hz),7.32(2H,t,J=7.6Hz),7.29(2H,d,J=7.2Hz),7.20(1H,t,J=7.6Hz),7.14(2H,d,J=8.4Hz),7.06(1H,t,J=7.6Hz),5.68(1H,dd,J=14.6,7.2Hz),5.21(1H,dd,J=13.6,7.6Hz),3.79(1H,dd,J=14.6,7.2Hz),3.64(1H,dd,J=14.6,7.2Hz),3.58(3H,s),3.24(1H,dd,J=13.6,7.6Hz),3.10(1H,dd,J=13.6,7.6Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.81 (1H, s, NH), 9.62 (1H, d, J = 8.0 Hz, NH), 9.23 (1H, d, J = 8.0 Hz, NH), 8.11 (2H, d, J = 8.0 Hz), 7.76 (1H, d, J = 7.6 Hz), 7.49 (1H, d, J = 7.6 Hz), 7.45 (1H, d, J = 1.6 Hz), 7.39 ( 1H, t, J = 7.6 Hz), 7.32 (2H, t, J = 7.6 Hz), 7.29 (2H, d, J = 7.2 Hz), 7.20 (1H, t, J = 7.6 Hz), 7.14 (2H, d, J = 8.4 Hz), 7.06 (1H, t, J = 7.6 Hz), 5.68 (1H, dd, J = 14.6, 7.2 Hz), 5.21 (1H, dd, J = 13.6, 7.6 Hz), 3.79 ( 1H, dd, J = 14.6, 7.2 Hz), 3.64 (1H, dd, J = 14.6, 7.2 Hz), 3.58 (3H, s), 3.24 (1H, dd, J = 13.6, 7.6 Hz), 3.10 (1H) , dd, J =13.6, 7.6 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),172.1(s),168.0(s),137.5(s),136.8(s),135.5(s),131.8(2×C,d),131.7(2×C,d),131.5(d),128.6(s),128.6(s),128.6(2×C,d),128.1(2×C,d),124.4(d),121.7(d),120.9(s),119.2(d),119.1(d),111.9(d),111.2(s),55.2(d),54.2(d),52.0(q),37.4(t),28.6(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.8 (s), 172.1 (s), 168.0 (s), 137.5 (s), 136.8 (s), 135.5 (s), 131.8 (2 × C, d ), 131.7 (2 × C, d), 131.5 (d), 128.6 (s), 128.6 (s), 128.6 (2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.7 (d), 120.9(s), 119.2(d), 119.1(d), 111.9(d), 111.2(s), 55.2(d), 54.2(d), 52.0(q), 37.4(t), 28.6 (t).

低解析噴灑式質譜(m/z，%)：570[M+Na]⁺(100),572[M+2+Na]⁺(100)。高解析度質譜：m/z 570.1007[M+Na]⁺(calcd for C₂₈H₂₆BrN₃O₄Na 570.1004)。 Low resolution spray mass spectrometry ( m/z , %): 570 [M+Na] ⁺ (100), 572 [M+2+Na] ⁺ (100). High-resolution mass spectrum: m/z 570.1007 [M+Na] ⁺ (calcd for C ₂₈ H ₂₆ BrN ₃ O ₄ Na 570.1004).

N-(N-苄基-L-色氨醯基)-對-三氟甲基-D-苯丙氨酸甲酯( N-(N-Benzoyl-L-tryptophanyl)-para-trifluoromethyl-D-phenylalanine methyl esters ，化合物23a和23b) N- ( N -benzyl -L-chromomethyl)-p-trifluoromethyl-D -phenylalanine methyl ester ( N-(N-Benzoyl-L-tryptophanyl)-para-trifluoromethyl-D- Phenylalanine methyl esters , compounds 23a and 23b)

化合物23a：產率26%。白色粉狀固體，熔點：233-235℃。 Compound 23a: Yield 26%. White powdery solid, melting point: 233-235 ° C.

¹H NMR(C₅D₅N)：δ11.88(1H,s,NH),9.50(1H,d,J=8.0Hz,NH),9.18(1H,d,J=8.0Hz,NH),8.08(2H,d,J=7.6Hz),7.89(1H,d,J=7.6Hz),7.54(1H,d,J=7.6Hz),7.45(2H,d,J=8.0Hz),7.42(1H,d,J=2.0Hz),7.36(1H,t,J=7.6Hz),7.29-7.23(3H,m),7.19(2H,d,J=8.0Hz),7.15 (1H,t,J=7.6Hz),5.67(1H,dd,J=14.4,7.2Hz),5.23(1H,dd,J=14.0,7.2Hz),3.79(1H,dd,J=14.4,7.2Hz),3.63(1H,dd,J=14.4,7.2Hz),3.52(3H,s),3.20(1H,dd,J=14.0,7.2Hz),3.14(1H,dd,J=14.0,7.2Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.88 (1H, s, NH), 9.50 (1H, d, J = 8.0 Hz, NH), 9.18 (1H, d, J = 8.0 Hz, NH), 8.08 (2H, d, J = 7.6 Hz), 7.89 (1H, d, J = 7.6 Hz), 7.54 (1H, d, J = 7.6 Hz), 7.45 (2H, d, J = 8.0 Hz), 7.42 ( 1H,d, J =2.0 Hz), 7.36 (1H, t, J = 7.6 Hz), 7.29-7.23 (3H, m), 7.19 (2H, d, J = 8.0 Hz), 7.15 (1H, t, J =7.6 Hz), 5.67 (1H, dd, J = 14.4, 7.2 Hz), 5.23 (1H, dd, J = 14.0, 7.2 Hz), 3.79 (1H, dd, J = 14.4, 7.2 Hz), 3.63 (1H) , dd, J = 14.4, 7.2 Hz), 3.52 (3H, s), 3.20 (1H, dd, J = 14.0, 7.2 Hz), 3.14 (1H, dd, J = 14.0, 7.2 Hz).

¹³C NMR(C₅D₅N)：δ172.8(s),171.9(s),167.6(s),142.1(s),137.5(s),135.4(s),131.5(d),130.4(2×C,d),128.7(2×C,d),128.6(s,J _C-F=32.2Hz),128.1(2×C,d),125.5(2×C,d,J _C-F=4.0Hz),125.0(s,J _C-F=270.1Hz),124.4(d),121.9(d),119.3(2×C,d),112.0(d),111.3(s),55.4(d),54.0(d),52.0(q),37.5(t),29.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.8 (s), 171.9 (s), 167.6 (s), 142.1 (s), 137.5 (s), 135.4 (s), 131.5 (d), 130.4 ( 2 × C, d), 128.7 (2 × C, d), 128.6 (s, J _CF = 32.2 Hz), 128.1 (2 × C, d), 125.5 (2 × C, d, J _CF = 4.0 Hz) , 125.0 (s, J _CF = 270.1 Hz), 124.4 (d), 121.9 (d), 119.3 (2 × C, d), 112.0 (d), 111.3 (s), 55.4 (d), 54.0 (d) , 52.0 (q), 37.5 (t), 29.0 (t).

低解析噴灑式質譜(m/z，%)：560[M+Na]⁺(100)。高解析度質譜：m/z 560.1776[M+Na]⁺(calcd for C₂₉H₂₆F₃N₃O₄Na 560.1773)。 Low analytical spray mass spectrometry ( m/z , %): 560 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 560.1776 [M+Na] ⁺ (calcd for C ₂₉ H ₂₆ F ₃ N ₃ O ₄ Na 560.1773).

化合物23b：產率9%。白色粉狀固體，熔點：187-189℃。 Compound 23b: Yield 9%. White powdery solid, melting point: 187-189 ° C.

¹H NMR(C₅D₅N)：δ11.81(1H,s,NH),9.72(1H,d,J=8.0Hz,NH),9.25(1H,d,J=8.0Hz,NH),8.12(2H,d,J=8.0Hz),7.75(1H,d,J=8.0Hz),7.49(1H,d,J=8.0Hz),7.44(1H,d,J=2.0Hz),7.43(2H,d,J=8.0Hz),7.40-7.37(3H,m),7.31(2H,d,J=8.0Hz),7.20(1H,t,J=8.0Hz),7.05(1H,t,J=8.0Hz),5.67(1H,dd,J=14.4,7.2Hz),5.25(1H,dd,J=14.0,7.2Hz),3.78(1H,dd,J=14.4,7.2Hz),3.62(1H,dd,J=14.4,7.2Hz),3.60(3H,s),3.36(1H,dd,J=13.6,8.0Hz),3.21(1H,dd,J=13.6,8.0Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.81 (1H, s, NH), 9.72 (1H, d, J = 8.0 Hz, NH), 9.25 (1H, d, J = 8.0 Hz, NH), 8.12 (2H, d, J = 8.0 Hz), 7.75 (1H, d, J = 8.0 Hz), 7.49 (1H, d, J = 8.0 Hz), 7.44 (1H, d, J = 2.0 Hz), 7.43 ( 2H, d, J = 8.0 Hz), 7.40-7.37 (3H, m), 7.31 (2H, d, J = 8.0 Hz), 7.20 (1H, t, J = 8.0 Hz), 7.05 (1H, t, J = 8.0 Hz), 5.67 (1H, dd, J = 14.4, 7.2 Hz), 5.25 (1H, dd, J = 14.0, 7.2 Hz), 3.78 (1H, dd, J = 14.4, 7.2 Hz), 3.62 (1H) , dd, J = 14.4, 7.2 Hz), 3.60 (3H, s), 3.36 (1H, dd, J = 13.6, 8.0 Hz), 3.21 (1H, dd, J = 13.6, 8.0 Hz).

¹³C NMR(C₅D₅N)：δ172.9(s),172.0(s),168.0(s),142.2(s),137.5(s),135.4(s),131.5(d),130.5(2×C,d),128.6(2×C,d),128.5(s,J _C-F=32.0Hz),128.1(2×C,d),125.5(2×C,d,J _C-F=4.0Hz),124.9(s,J _C-F=270.0Hz),124.4(d),121.9(d),119.2(d),119.1(d),111.9(d),111.1(s),55.4(d),54.0(d),52.0(q),37.5(t),29.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.9 (s), 172.0 (s), 168.0 (s), 142.2 (s), 137.5 (s), 135.4 (s), 131.5 (d), 130.5 ( 2 × C, d), 128.6 (2 × C, d), 128.5 (s, J _CF = 32.0 Hz), 128.1 (2 × C, d), 125.5 (2 × C, d, J _CF = 4.0 Hz) , 124.9 (s, J _CF = 270.0 Hz), 124.4 (d), 121.9 (d), 119.2 (d), 119.1 (d), 111.9 (d), 111.1 (s), 55.4 (d), 54.0 (d ), 52.0 (q), 37.5 (t), 29.0 (t).

低解析噴灑式質譜(m/z，%)：560[M+Na]⁺(100)。高解析度質譜：m/z 560.1775[M+Na]⁺(calcd for C₂₉H₂₆F₃N₃O₄Na 560.1773)。 Low analytical spray mass spectrometry ( m/z , %): 560 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 560.1775 [M+Na] ⁺ (calcd for C ₂₉ H ₂₆ F ₃ N ₃ O ₄ Na 560.1773).

N-(N-苄基-L-色氨醯基)-3-環己基-D-丙氨酸甲酯( N-(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters ，化合物24a和24b) N - (N - benzyl-acyl -L- tryptophyl) -3-cyclohexyl-alanine methyl ester -D- (N- (N-benzoyl-L -tryptophanyl) -3-cyclohexyl-D-alanine methyl esters , compounds 24a and 24b)

化合物24a：產率39%。白色粉狀固體，熔點：170-172℃。 Compound 24a: Yield 39%. White powdery solid, melting point: 170-172 ° C.

¹H NMR(C₅D₅N)：δ11.88(1H,s,NH),9.45(1H,d,J=8.0Hz,NH),9.07(1H,d,J=8.0Hz,NH),8.08(2H,d,J=8.4Hz),7.94(1H,d,J=7.6Hz),7.56(1H,d,J=7.6Hz),7.50(1H,br.s),7.35(1H,dt,J=7.6,1.2Hz),7.29-7.23(3H,m),7.14(1H,t,J=7.6Hz),5.72(1H,dd,J=14.2,7.2Hz),5.00(1H,m),3.86(1H,dd,J=14.2,7.2Hz),3.72(1H,dd,J=14.2,7.2Hz),3.58(3H,s),1.77(1H,m),1.66(2H,m),1.59-1.48(4H,m),1.38(1H,m),1.25-0.96(3H,m),0.84-0.71(2H,m)。 ¹ H NMR (C ₅ D ₅ N): δ 11.88 (1H, s, NH), 9.45 (1H, d, J = 8.0 Hz, NH), 9.07 (1H, d, J = 8.0 Hz, NH), 8.08 (2H, d, J = 8.4 Hz), 7.94 (1H, d, J = 7.6 Hz), 7.56 (1H, d, J = 7.6 Hz), 7.50 (1H, br.s), 7.35 (1H, dt) , J = 7.6, 1.2 Hz), 7.29-7.23 (3H, m), 7.14 (1H, t, J = 7.6 Hz), 5.72 (1H, dd, J = 14.2, 7.2 Hz), 5.00 (1H, m) , 3.86 (1H, dd, J = 14.2, 7.2 Hz), 3.72 (1H, dd, J = 14.2, 7.2 Hz), 3.58 (3H, s), 1.77 (1H, m), 1.66 (2H, m), 1.59-1.48 (4H, m), 1.38 (1H, m), 1.25-0.96 (3H, m), 0.84-0.71 (2H, m).

¹³C NMR(C₅D₅N)：δ173.7(s),172.9(s),167.7(s),137.5(s),135.5(s),131.4(s),128.7(s),128.5(2×C,d),128.0(2×C,d),124.3(d),121.8(d),119.3(d),119.2(d),112.0(d),111.2(d),55.4(d),51.9(q),50.9(d),39.4(t),34.1(d),33.6(t),32.4(t),29.2(t),26.6(t),26.3(t),26.1(t)。 ¹³ C NMR (C ₅ D ₅ N): δ 173.7 (s), 172.9 (s), 167.7 (s), 137.5 (s), 135.5 (s), 131.4 (s), 128.7 (s), 128.5 ( 2 × C, d), 128.0 (2 × C, d), 124.3 (d), 121.8 (d), 119.3 (d), 119.2 (d), 112.0 (d), 111.2 (d), 55.4 (d) , 51.9(q), 50.9(d), 39.4(t), 34.1(d), 33.6(t), 32.4(t), 29.2(t), 26.6(t), 26.3(t), 26.1(t) .

低解析噴灑式質譜(m/z，%)：498[M+Na]⁺(100)。高解析度質譜：m/z 498.2366[M+Na]⁺(calcd for C₂₈H₃₃N₃O₄Na 498.2369)。 Low analytical spray mass spectrometry ( m/z , %): 498 [M+Na] ⁺ (100). High Resolution Mass Spectrum: m / z 498.2366 [M + Na] + (calcd for C 28 H 33 N 3 O 4 Na 498.2369).

化合物24b：產率14%。白色粉狀固體，熔點：109-111℃。 Compound 24b: Yield 14%. White powdery solid, melting point: 109-111 ° C.

¹H NMR(C₅D₅N)：δ11.82(1H,s,NH),9.67(1H,d,J=8.0Hz,NH),9.27(1H,d,J=8.0Hz,NH),8.12(2H,d,J=8.0Hz),7.78(1H,d,J=7.6Hz),7.55(1H,br.s),7.49(1H,d,J=7.6Hz),7.35(1H,dt,J=1.2,8.0Hz),7.28(2H,dt,J=1.2,8.0Hz),7.20(1H,t,J=7.6Hz),7.06(1H,t,J=7.6Hz),5.71(1H,dd,J=14.6,7.6Hz),5.12(1H,dd,J=14.6,7.6Hz),3.86(1H,dd,J=14.6,7.2Hz),3.71(1H,dd,J=14.6,7.2Hz),3.65(3H,s),1.73(2H,m),1.63(1H,m),1.54-1.41(5H,m),1.13-0.92(3H,m),0.85-0.70(2H,m)。 ¹ H NMR (C ₅ D ₅ N): δ 11.82 (1H, s, NH), 9.67 (1H, d, J = 8.0 Hz, NH), 9.27 (1H, d, J = 8.0 Hz, NH), 8.12 (2H, d, J = 8.0 Hz), 7.78 (1H, d, J = 7.6 Hz), 7.55 (1H, br.s), 7.49 (1H, d, J = 7.6 Hz), 7.35 (1H, dt , J = 1.2, 8.0 Hz), 7.28 (2H, dt, J = 1.2, 8.0 Hz), 7.20 (1H, t, J = 7.6 Hz), 7.06 (1H, t, J = 7.6 Hz), 5.71 (1H) , dd, J = 14.6, 7.6 Hz), 5.12 (1H, dd, J = 14.6, 7.6 Hz), 3.86 (1H, dd, J = 14.6, 7.2 Hz), 3.71 (1H, dd, J = 14.6, 7.2 Hz), 3.65 (3H, s), 1.73 (2H, m), 1.63 (1H, m), 1.54-1.41 (5H, m), 1.13-0.92 (3H, m), 0.85-0.70 (2H, m) .

¹³C NMR(C₅D₅N)：δ173.9(s),173.0(s),167.8(s),137.5(s),135.5(s),131.4(s),128.7(s),128.5(2×C,d),128.1(2×C,d),124.4(d),121.7(d),119.2(2×C,d),111.9(d),111.2(d),55.3(d),52.0(q),50.8(d),39.7(t),34.2(d),33.7(t),32.3(t),28.6(t),26.6(t),26.3(t),26.1(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ173.9 (s), 173.0 (s), 167.8 (s), 137.5 (s), 135.5 (s), 131.4 (s), 128.7 (s), 128.5 ( 2 × C, d), 128.1 (2 × C, d), 124.4 (d), 121.7 (d), 119.2 (2 × C, d), 111.9 (d), 111.2 (d), 55.3 (d), 52.0(q), 50.8(d), 39.7(t), 34.2(d), 33.7(t), 32.3(t), 28.6(t), 26.6(t), 26.3(t), 26.1(t).

低解析噴灑式質譜(m/z，%)：498[M+Na]⁺(100)。高解析度質譜：m/z 498.2367[M+Na]⁺(calcd for C₂₈H₃₃N₃O₄Na 498.2369)。 Low analytical spray mass spectrometry ( m/z , %): 498 [M+Na] ⁺ (100). High-resolution mass spectrum: m/z 498.2367 [M+Na] ⁺ (calcd for C ₂₈ H ₃₃ N ₃ O ₄ Na 498.2369).

【實施例五】N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸氨基乙腈( N-(N-benzoyl-L-tryptophanyl)-D-phenylalanine-glycine-nitriles，化合物27a與27b)的製備方法如下： [Fifth Embodiment] N - (N - aralkyl acyl -L- tryptophyl) -D- phenylalanine aminoacetonitrile (N- (N-benzoyl-L -tryptophanyl) -D-phenylalanine-glycine-nitriles The preparation methods of the compounds 27a and 27b) are as follows:

二碳酸二叔丁酯-D-苯丙氨酸(Boc-D-phenyl-alanine)(1.0當量)與氨基乙腈(aminoacetonitrile)(1.0當量)進行反應，以二氯甲烷(CH₂Cl₂)為溶媒，隨後加入苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)(2.0當量)及N,N-二異丙基乙胺(DIEA)(1.5當量)在室溫下反應6小時，經管柱層析進行純化〔溶媒系統乙酸乙酯(EtOAc)/正己烷(n-hexane)=4/6〕，得到第一個中間產物Boc-D-phenylalanyl-glycine-nitrile。接著加入20%三氟乙酸(TFA)(TFA/CH₂Cl₂=1/4)進行切除二碳酸二叔丁酯(Boc)保護基團，室溫下反應30分鐘，經管柱層析進行純化〔溶媒系統甲醇(MeOH)/三氯甲烷(CHCl₃)=1/10〕，得到D-phenylalanyl-glycine-nitrile。N-benzoyl-L-tryptophan(1.0當量)與N-benzoyl-5-hydroxy-L-tryptophan(1.0當量)各別D-phenylalaninol(D-2-amino-3-phenyl-1-propanol)(1.0當量)及D-phenylalanyl-glycine-nitrile(1.0當量)進行反應，以CH₂Cl₂為溶媒，隨後加入苯幷三氮唑-N,N,N',N'-四甲基脲六氟磷酸鹽(HBTU)(2.0當量)及N,N-二異丙基乙胺(DIEA)(1.5當量)在室溫下反應6小時，經以溶媒系統正已烷/乙酸乙酯(1：1)進行管柱層析純化，再經過高效液相色譜法(HPLC)分離純化(溶媒系統：65% MeOH+0.3% TFA)，即可得到最終產物(化合物27a與27b)。 Bic-D-phenyl-alanine (1.0 eq.) was reacted with aminoacetonitrile (1.0 eq.) with dichloromethane (CH ₂ Cl ₂ ). Solvent, followed by the addition of benzotriazole- N,N,N',N' -tetramethyluronium hexafluorophosphate (HBTU) (2.0 equivalents) and N,N-diisopropylethylamine (DIEA) ( The reaction was carried out for 6 hours at room temperature, and purified by column chromatography [solvent system ethyl acetate (EtOAc) / n-hexane (n-hexane) = 4/6] to give the first intermediate product Boc-D- Phenylalanyl-glycine-nitrile. Then, 20% trifluoroacetic acid (TFA) (TFA/CH ₂ Cl ₂ = 1/4) was added to remove the di-tert-butyl dicarbonate (Boc) protecting group, and the reaction was carried out for 30 minutes at room temperature, and purified by column chromatography. [Solvent system methanol (MeOH) / chloroform (CHCl ₃ ) = 1/10] gave D-phenylalanyl-glycine-nitrile. N- benzoyl-L-tryptophan (1.0 eq.) and N- benzoyl-5-hydroxy-L-tryptophan (1.0 eq.), respectively, D-phenylalaninol (D-2-amino-3-phenyl-1-propanol) (1.0 equivalents) And D-phenylalanyl-glycine-nitrile (1.0 eq.), using CH ₂ Cl ₂ as a solvent, followed by the addition of benzotriazole- N,N,N',N' -tetramethyluronium hexafluorophosphate (HBTU) (2.0 eq.) and N,N-diisopropylethylamine (DIEA) (1.5 eq.) were allowed to react at room temperature for 6 hours, and the solvent system n-hexane/ethyl acetate (1:1) Purification by column chromatography and purification by high performance liquid chromatography (HPLC) (solvent system: 65% MeOH + 0.3% TFA) gave the final product ( compounds 27a and 27b ).

化合物27a：產率49%。白色粉狀固體，熔點：167-169℃。 Compound 27a: Yield 49%. White powdery solid, melting point: 167-169 °C.

¹H NMR(C₅D₅N)：δ1.81(1H,s,NH),9.73(1H,t,J=5.6Hz,NH),9.63(1H,d,J=8.4Hz,NH),8.39(1H,d,J=6.4Hz,NH),8.07(2H,d,J=7.6Hz),7.77(1H,d,J=8.0Hz),7.56(1H,d,J=8.0Hz),7.35(1H,t,J=7.6Hz),7.29-7.23(5H,m),7.20-7.18(3H,m),7.15-7.09(2H,m),5.35-5.29(2H,m),5.00(1H,m),4.47(2H,d,J=5.6Hz),3.70(1H,dd,J=14.2,7.6Hz),3.61(1H,dd,J=14.2,7.6Hz),3.33(1H,dd,J=13.6,8.0Hz),3.09(1H,dd,J=13.6,8.0Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 1.81 (1H, s, NH), 9.73 (1H, t, J = 5.6 Hz, NH), 9.63 (1H, d, J = 8.4 Hz, NH), 8.39 (1H, d, J = 6.4 Hz, NH), 8.07 (2H, d, J = 7.6 Hz), 7.77 (1H, d, J = 8.0 Hz), 7.56 (1H, d, J = 8.0 Hz), 7.35 (1H, t, J = 7.6 Hz), 7.29-7.23 (5H, m), 7.20-7.18 (3H, m), 7.15-7.09 (2H, m), 5.35-5.29 (2H, m), 5.00 ( 1H, m), 4.47 (2H, d, J = 5.6 Hz), 3.70 (1H, dd, J = 14.2, 7.6 Hz), 3.61 (1H, dd, J = 14.2, 7.6 Hz), 3.33 (1H, dd , J = 13.6, 8.0 Hz), 3.09 (1H, dd, J = 13.6, 8.0 Hz).

¹³C NMR(C₅D₅N)：δ173.2(s),172.7(s),166.8(s),138.1(s),137.5(s),135.5(s),131.7(s),129.7(2×C,d),128.8(2×C,d),128.5(s),128.5(2×C,d),128.2(2×C,d),126.9(d),124.5(d),121.9(d),119.3(d),119.1(d),117.6(s),112.0(s),110.9(s),56.6(d),55.0(d),37.8(t),28.4(t),28.0(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ173.2 (s), 172.7 (s), 166.8 (s), 138.1 (s), 137.5 (s), 135.5 (s), 131.7 (s), 129.7 ( 2 × C, d), 128.8 (2 × C, d), 128.5 (s), 128.5 (2 × C, d), 128.2 (2 × C, d), 126.9 (d), 124.5 (d), 121.9 (d), 119.3(d), 119.1(d), 117.6(s), 112.0(s), 110.9(s), 56.6(d), 55.0(d), 37.8(t), 28.4(t), 28.0 (t).

低解析噴灑式質譜(m/z，%)：516[M+Na]⁺(100)。高解析度質譜：m/z 516.2008[M+Na]⁺(calcd for C₂₉H₂₇N₅O₃Na 516.2011)。 Low analytical spray mass spectrometry ( m/z , %): 516 [M+Na] ⁺ (100). High resolution mass spectrum: m/z 516.2008 [M+Na] ⁺ (calcd for C ₂₉ H ₂₇ N ₅ O ₃ Na 516.2011).

化合物27b：產率7%。白色粉狀固體，熔點：195-197℃。 Compound 27b: Yield 7%. White powdery solid, melting point: 195-197 ° C.

¹H NMR(C₅D₅N)：δ11.80(1H,s,NH),9.76(1H,d,J=8.0Hz,NH),9.63(1H,t,J=5.2Hz,NH),9.10(1H,d,J=7.6Hz,NH),8.07(2H,d,J=8.0Hz),7.74(1H,d,J=8.0Hz),7.49(1H,d,J=8.0Hz),7.44(1H,br.s),7.37(1H,t,J=7.2Hz),7.31-7.25(4H,m),7.18-7.14(3H,m),7.10-7.04(2H,m),5.59(1H,dd,J=14.2,7.2Hz),5.18(1H,dd,J=14.2,7.2Hz),4.43(1H,dd,J=17.2,5.6Hz),4.32(1H,dd,J=17.2,5.6Hz),3.74(1H,dd,J=14.2,7.6Hz),3.62(1H,dd,J=14.2,7.6Hz),3.41(1H,dd,J=13.6,8.0Hz),3.18(1H,dd,J=13.6,8.0Hz)。 ¹ H NMR (C ₅ D ₅ N): δ 11.80 (1H, s, NH), 9.76 (1H, d, J = 8.0 Hz, NH), 9.63 (1H, t, J = 5.2 Hz, NH), 9.10 (1H, d, J = 7.6 Hz, NH), 8.07 (2H, d, J = 8.0 Hz), 7.74 (1H, d, J = 8.0 Hz), 7.49 (1H, d, J = 8.0 Hz), 7.44 (1H, br.s), 7.37 (1H, t, J = 7.2 Hz), 7.31-7.25 (4H, m), 7.18-7.14 (3H, m), 7.10-7.04 (2H, m), 5.59 ( 1H, dd, J = 14.2, 7.2 Hz), 5.18 (1H, dd, J = 14.2, 7.2 Hz), 4.43 (1H, dd, J = 11.2, 5.6 Hz), 4.32 (1H, dd, J = 17.2, 5.6 Hz), 3.74 (1H, dd, J = 14.2, 7.6 Hz), 3.62 (1H, dd, J = 14.2, 7.6 Hz), 3.41 (1H, dd, J = 13.6, 8.0 Hz), 3.18 (1H, Dd, J = 13.6, 8.0 Hz).

¹³C NMR(C₅D₅N)：δ172.7(s),172.5(s),168.0(s),137.9(s),137.4(s),135.5(s),131.5(s),129.8(2×C,d),128.8(2×C,d),128.6(s),128.5(2×C,d),128.1(2×C,d),126.9(d),124.5(d),121.7(d),119.2(d),119.1(d),117.5(s),112.0(s),111.0(s),55.4(d),55.3(d),38.4(t),28.5(t),27.8(t)。 ^{_{_{13 C NMR (C 5 D 5}}} N): δ172.7 (s), 172.5 (s), 168.0 (s), 137.9 (s), 137.4 (s), 135.5 (s), 131.5 (s), 129.8 ( 2 × C, d), 128.8 (2 × C, d), 128.6 (s), 128.5 (2 × C, d), 128.1 (2 × C, d), 126.9 (d), 124.5 (d), 121.7 (d), 119.2(d), 119.1(d), 117.5(s), 112.0(s), 111.0(s), 55.4(d), 55.3(d), 38.4(t), 28.5(t), 27.8 (t).

【實施例六】人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成抑制活性評估 [Example 6] Evaluation of human neutrophil elastase release and superoxide radical production inhibition activity

人類嗜中性白血球的製備：取自年約20-30歲作息正常且禁服藥物達兩周以上之健康捐血者，以含有肝素(heparin)的真空採血管於肘靜脈採血。在室溫下以650g離心8或10分鐘後，移除上層液，再加入等體積3%右旋糖酐(dextran)溶液混合均勻，並於室溫靜置30分鐘。取出含有嗜中性白血球之上層懸浮液加入單核球細胞分離液 (Ficoll-Paque)，於20℃下以400g離心40分鐘。以低張溶液將下層小丸(pellet)中殘餘紅血球脹破，再以4℃、200g離心8分鐘去除上清液，最後將分離出之嗜中性白血球重新懸浮於不含鈣的平衡鹽溶液(HBSS)中進行實驗。 Preparation of human neutrophils: Take a healthy blood donor with a normal work schedule of about 20-30 years old and banned medication for more than two weeks, and collect blood in the elbow vein with a vacuum blood collection tube containing heparin. After centrifugation at 650 g for 8 or 10 minutes at room temperature, the supernatant was removed, and an equal volume of 3% dextran solution was added and mixed uniformly, and allowed to stand at room temperature for 30 minutes. Remove the supernatant containing neutrophils from the supernatant and add mononuclear cell fraction (Ficoll-Paque), centrifuged at 400 g for 40 minutes at 20 °C. The residual red blood cells in the lower pellet were swelled with a low-tension solution, and then the supernatant was removed by centrifugation at 200 ° C for 8 minutes at 4 ° C, and finally the separated neutrophils were resuspended in a balanced salt solution containing no calcium ( Experiments were performed in HBSS).

超氧自由基(Superoxide,O₂ ^‧-)的釋放測定：由於正鐵細胞色素C(ferricytochrome c)無法穿透細胞膜，在細胞外與超氧自由基反應，使細胞色素c還原，且於波長550nm時有吸光反應，以測量吸光值變化的方式來評估待測化合物對超氧自由基釋放的影響。將含有0.5mg/ml細胞色素c的嗜中性白血球懸浮液(6×10⁵cells/ml)，於37℃與待測化合物作用2分鐘，再加入1μg/ml細胞鬆弛素B(cytochalasin B)作用3分鐘來放大反應，最後加入fMLF(30nM)或WKYMVm(1.5nM)反應10分鐘。使用分光光度計於波長550nm下測量吸光值變化。 Superoxide (O ₂ ^‧- ) release assay: Since ferricytochrome c cannot penetrate the cell membrane, it reacts with superoxide radicals outside the cell to reduce cytochrome c and at wavelength There is a light absorption reaction at 550 nm, and the effect of the test compound on the release of superoxide radicals is evaluated by measuring the change in absorbance. A neutrophil suspension (6 × 10 ⁵ cells / ml) containing 0.5 mg / ml of cytochrome c was applied to the test compound for 2 minutes at 37 ° C, and then 1 μg / ml of cytochalasin B was added. The reaction was amplified for 3 minutes and finally reacted with fMLF (30 nM) or WKYMVm (1.5 nM) for 10 minutes. The change in absorbance was measured using a spectrophotometer at a wavelength of 550 nm.

【實施例七】細胞毒性測試 [Example 7] Cytotoxicity test

在圖1(a)~圖1(b)繪示HCH6-1對於人類嗜中性白血球釋放乳酸脫氫酶(lactate dehydrogenase,LDH)影響，其實驗條件如下。將嗜中性白血球懸浮液於37℃與不同濃度待測化合物反應15分鐘或固定濃度反應不同時間，另以0.1% Triton X-100反應30分鐘作為總乳酸脫氫酶(Total LDH)釋放量，於4℃、200g離心8分鐘取上清液，加入乳酸脫氫酶試劑於室溫下避光反應15分鐘，於波長492nm下測量吸光值。結果以總乳酸脫氫酶釋放量為100%來計算。 Fig. 1(a) to Fig. 1(b) show the effect of HCH6-1 on the release of lactate dehydrogenase (LDH) from human neutrophils, and the experimental conditions are as follows. The neutrophil suspension was reacted with different concentrations of the test compound at 37 ° C for 15 minutes or fixed concentration for different time, and further reacted with 0.1% Triton X-100 for 30 minutes as total lactate dehydrogenase (Total LDH) release. The supernatant was centrifuged at 200 g for 8 minutes at 4 ° C, and the lactate dehydrogenase reagent was added thereto to protect from light at room temperature for 15 minutes, and the absorbance was measured at a wavelength of 492 nm. The results were calculated as 100% of total lactate dehydrogenase release.

圖1(a)實驗結果顯示HCH6-1抑制fMLF刺激人類嗜中性白血球釋放超氧自由基和彈性蛋白酶的作用，並不是透過毒殺嗜中性白血球所導致的。 Figure 1 (a) experimental results show that HCH6-1 inhibits the release of superoxide radicals and elastase by human neutrophils induced by fMLF, not by poisoning neutrophils.

圖1(b)顯示：在長時間的狀態下，HCH6-1 30μM與細胞作用至120分鐘仍然不具有顯著的毒性，證明高濃度的HCH6-1與細胞作用120分鐘內是無毒殺細胞的能力。 Figure 1 (b) shows that under long-term conditions, HCH6-1 30μM and cells for 120 minutes still have no significant toxicity, demonstrating the ability of high concentrations of HCH6-1 to act as a non-toxic cell killer within 120 minutes of cell action. .

【實施例八】彈性蛋白酶活性測定 [Example 8] Determination of elastase activity

將含有1mM鈣離子的嗜中性白血球懸浮液，於37℃ 下以0.5g/ml細胞鬆弛素B作用2分鐘，最後加入fMLF反應15分鐘後，以4℃、1000g離心5分鐘取上清液。上清液於37℃下與待測化合物作用2分鐘，再加入N-甲氧基琥珀醯-丙醯氨-丙醯氨-脯醯氨-纈氨酸對硝基醯苯胺(N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide)100M反應10分鐘。使用分光光度計於波長405nm下測量吸光值變化。 The neutrophil suspension containing 1 mM calcium ion was treated with 0.5 g/ml cytochalasin B for 2 minutes at 37 ° C, and finally added to the fMLF reaction for 15 minutes, and then centrifuged at 1000 ° for 5 minutes at 4 ° C for 5 minutes. . The supernatant was allowed to react with the test compound for 2 minutes at 37 ° C, and then N -methoxysuccinyl-propionamidine-propionamidine-ammonium-valine p-nitroanilide ( N- methoxysuccinyl- Ala-Ala-Pro-Val- p- nitroanilide) was reacted at 100 M for 10 minutes. The change in absorbance was measured using a spectrophotometer at a wavelength of 405 nm.

【實施例九】細胞表面CD11b的表現將嗜中性白血球懸浮液在37℃下混勻，預熱5分鐘後加入不同濃度待測化合物，接著加入細胞鬆弛素B(0.5g/ml)反應3分鐘再加入fMLF(30nM)反應10分鐘後置冰上終止反應。離心後去除上清液，將細胞重新懸浮於含5%牛血清白蛋白(BSA)的平衡鹽溶液中，加入異硫氰酸螢光素(FITC)標定的anti-CD11b於冰上避光反應，最後再加入平衡鹽溶液終止反應。利用具異硫氰酸螢光素標定的anti-CD11b，以流式細胞儀來偵測，根據螢光表現量的變化來進一步了解待測化合物是否會影響，受刺激後嗜中性白血球細胞膜上CD11b的表現。結果顯示，在fMLF刺激下，HCH6-1具有濃度相關性的抑制細胞膜上的CD11b的表現，IC₅₀為0.26±0.05μM(請參閱圖2(a)和圖2(b))，然而在Leu-Glu-Ser-Ile-Phe-Arg-Ser-Leu-Leu-Phe-Arg-Val-Met(MMK1)及IL-8刺激之下，HCH6-1並無抑制的作用(請參閱圖2(c)~圖2(f))。 [Example 9] Expression of cell surface CD11b The neutrophil suspension was mixed at 37 ° C, and after preheating for 5 minutes, different concentrations of the test compound were added, followed by the addition of cytochalasin B (0.5 g/ml). After a further 10 minutes of reaction with fMLF (30 nM), the reaction was stopped on ice. After centrifugation, the supernatant was removed, and the cells were resuspended in a balanced salt solution containing 5% bovine serum albumin (BSA), and anti-CD11b calibrated with fluorescein isothiocyanate (FITC) was added to protect against light on ice. Finally, a balanced salt solution is added to terminate the reaction. Using anti-CD11b calibrated with luciferin isothiocyanate, it is detected by flow cytometry. According to the change of fluorescence expression, it is further understood whether the test compound will affect the neutrophil cell membrane after stimulation. The performance of CD11b. The results showed that under the stimulation of fMLF, HCH6-1 had a concentration-dependent inhibition of CD11b expression on the cell membrane with an IC ₅₀ of 0.26 ± 0.05 μM (see Figure 2 (a) and Figure 2 (b)), however in Leu -Glu-Ser-Ile-Phe-Arg-Ser-Leu-Leu-Phe-Arg-Val-Met (MMK1) and IL-8 stimulation, HCH6-1 has no inhibitory effect (see Figure 2 (c ) ~ Figure 2 (f)).

【實施例十】受體測試 [Example 10] Receptor test

嗜中性白血球細胞膜上含有fMLF接受體(formyl peptide receptor 1,FPR1)，藉由會和FPR1結合且具備螢光性質的N-甲醯甲硫氨醯亮氨醯苯丙氨酸(Formyl-NIe-Leu-Phe-NIe-Tyr-Lys，FNLFNYK)來與待測化合物做接受體的競爭結合。因此以接受體結合測定，進一步釐清待測化合物是否和接受體作用。在圖3(a)~圖3(b)繪示HCH6-1抑制FNLFNYK結合在人類嗜中性白血球膜上的FPR1的效應，實驗結果顯示，HCH6-1具有濃度相關性的抑制FNLFNYK結合在人類嗜中性白血球膜上的FPR1，IC₅₀為2.02±0.34μM。 Neutrophil leukocyte cell membrane contains fMLF receptor (FPR1), which is a fluorescent-enhanced N-methionine leucine phenylalanine (Formyl-NIe) that binds to FPR1. -Leu-Phe-NIe-Tyr-Lys, FNLFNYK) to compete with the test compound as a receptor. Therefore, the receptor binding assay is used to further clarify whether or not the test compound acts on the acceptor. Figure 3(a) to Figure 3(b) show the effect of HCH6-1 on FPR1 binding of FNLFNYK to human neutrophils, and the results show that HCH6-1 has a concentration-dependent inhibition of FNLFNYK binding in humans. FPR1 on the neutrophil globule membrane had an IC ₅₀ of 2.02 ± 0.34 μM.

【實施例十一】西方墨點法 [Embodiment 11] Western ink dot method

Mitogen-activated protein kinase(MAPKs)和Akt的磷酸化表現與細胞發炎反應的調控息息相關。藉由專一性抗體的結合，觀察化合物對於不同訊息所激活的蛋白質表現量的影響。在嗜中性白血球中，MAPKs(ERK、p38和JNK)及Akt訊息傳遞路徑參與調控超氧自由基產生以及彈性蛋白酶釋放。並且於先前的實驗得知，HCH6-1具競爭性抑制超氧自由基產生以及彈性蛋白酶釋放。因此，欲探討FPR1下游中MAPKs及Akt訊息傳遞路徑是否參與HCH6-1競爭性抑制超氧自由基產生以及彈性蛋白酶釋放。 Mitogen-activated protein kinase (MAPKs) and Akt phosphoric acid The performance is closely related to the regulation of cellular inflammatory response. The effect of the compound on the amount of protein activated by different messages was observed by the binding of specific antibodies. In neutrophils, MAPKs (ERK, p38, and JNK) and the Akt signaling pathway are involved in the regulation of superoxide production and elastase release. And it was found in previous experiments that HCH6-1 competitively inhibits superoxide radical production and elastase release. Therefore, it is necessary to investigate whether MAPKs and Akt signaling pathways in the downstream of FPR1 are involved in the competitive inhibition of superoxide radical production and elastase release by HCH6-1.

圖4(a)~圖4(d)繪示HCH6-1具有顯著的抑制MAPKs(ERK、p38和JNK)及Akt磷酸化，其實驗條件如下：前處理HCH6-1(10μM)5分鐘，接著於fMLF(3-30nM)刺激反應30秒。實驗結果顯示，HCH6-1有抑制細胞內MAPKs及Akt的訊息傳遞，並且在低濃度fMLF(3nM)刺激之下，HCH6-1具有顯著的抑制MAPKs及Akt磷酸化(電泳呈現淺色帶)，間接證明其競爭性的能力(請參閱圖4(a)~圖4(d))。 Figure 4(a) to Figure 4(d) show that HCH6-1 has significant inhibition of MAPKs (ERK, p38 and JNK) and Akt phosphorylation, and the experimental conditions are as follows: pretreatment of HCH6-1 (10 μM) for 5 minutes, followed by The reaction was stimulated for 30 seconds at fMLF (3-30 nM). The results showed that HCH6-1 inhibited the signaling of intracellular MAPKs and Akt, and under the stimulation of low concentration of fMLF (3nM), HCH6-1 significantly inhibited MAPKs and Akt phosphorylation (electrophoresis showed a light band). The ability to indirectly prove its competitiveness (see Figure 4(a) ~ Figure 4(d)).

【實施例十二】統計學分析 [Twelfth embodiment] Statistical analysis

本實驗是採用GraphPad Prism軟體(GraphPad Software,San Diego,CA)來進行統計分析。所有的數據是藉由史徒登氏t-試驗(Student’s t-test)或單因子變異數分析(one-way analysis of variance,ANOVA)，並且繼之以特奇氏檢定(Turkey’s test)來作分析，俾以評估各組之間的差異性。若所得到的統計比對結果是p<0.05，則代表有統計學顯著性。 This experiment was performed using the GraphPad Prism software (GraphPad Software, San Diego, CA) for statistical analysis. All data history only by Gordon's t - test (Student's t -test), or a single factor analysis of variance (one-way analysis of variance, ANOVA), followed by Tec's assay and (Turkey's test) to make Analysis, to assess differences between groups. If the resulting statistical alignment result is p < 0.05, then it is statistically significant.

【實施例十三】：一種可治療一嗜中性白血球發炎性障礙的甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)拮抗劑，其包括如式一所示的化合物(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters)：其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；R₁選自氫(hydrogen atom)、羥基(hydroxyl group)和甲氧基(Methoxy)的其中之一；R₂選自無取代苯基(non-substituted phenyl group)、單取代苯基(mono-substituted phenyl group)、雙取代苯基(di-substituted phenyl group)、三取代苯基(tri-substituted phenyl group)、吡啶(pyridinyl)和C₄-C₆環烷基(cycloalkyl group)的其中之一；R₃選自無取代苯醯基團(non-substituted benzoyl group)、單取代苯醯基團(mono-substituted benzoyl group)、雙取代苯醯基團(di-substituted benzoyl group)和三取代苯醯基團(tri-substituted benzoyl group)的其中之一；以及R₄選自羥基(hydroxyl)、C₁-C₄烷氧基(alkoxyl)和甘氨酸氰(glycin-nitrile)的其中之一。 [Example 13]: A Formyl Peptide Receptor 1 (FPR1) antagonist capable of treating a neutrophil inflammatory disorder, comprising a compound as shown in Formula 1 ( N -( N -aroyl-L-tryptophanyl)-D-phenylalanine methyl esters): wherein: the chiral centers of the compound of the formula 1 are in the S configuration and the R configuration, respectively; and R _{1 is} selected from hydrogen ( One of hydrogen atom), a hydroxyl group, and a methoxy group; R _{2 is} selected from a non-substituted phenyl group, a mono-substituted phenyl group, and a double One of a di-substituted phenyl group, a tri-substituted phenyl group, a pyridinyl group, and a C ₄ -C ₆ cycloalkyl group; R _{3 is} selected from none a substituted non-substituted benzoyl group, a mono-substituted benzoyl group, a di-substituted benzoyl group, and a trisubstituted benzoquinone group (tri-) wherein one substituted benzoyl group); and R ₄ is selected from hydroxyl (hydroxyl), C ₁ -C ₄ alkoxy (Alkoxyl) and glycine cyanide (glycin-nitrile) is one of them.

【實施例十四】：一種具有抑制N-甲醯甲硫氨醯亮氨醯苯丙氨酸fMLF所誘導的人類嗜中性白血球彈性蛋白酶釋放與超氧自由基生成的可治療一嗜中性白血球發炎性障礙的甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)拮抗劑，其包括如式一所示的化合物(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters)：其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；R₁選自氫(hydrogen atom)、羥基(hydroxyl group)和甲氧基(Methoxy)的其中之一；R₂選自無取代苯基(non-substituted phenyl group)、單取代苯基(mono-substituted phenyl group)、雙取代苯基(di-substituted phenyl group)、三取代苯基(tri-substituted phenyl group)、吡啶(pyridinyl)和C₄-C₆環烷基(cycloalkyl group)的其中之一；R₃選自無取代苯醯基團(non-substituted benzoyl group)、單取代苯醯基團(mono-substituted benzoyl group)、雙取代苯醯基團(di-substituted benzoyl group)和三取代苯醯基團(tri-substituted benzoyl group)的其中之一；以及R₄選自羥基(hydroxyl)、C₁-C₄烷氧基(alkoxyl)和甘氨酸氰(glycin-nitrile)的其中之一。 [Embodiment 14]: A therapeutic neutrophil capable of inhibiting human neutrophil elastase release and superoxide radical production induced by inhibition of N -mercaptomethionine leucine phenylalanine fMLF A Formyl Peptide Receptor 1 (FPR1) antagonist comprising a compound of the formula ( N - ( N -aroyl-L-tryptophanyl)-D-) Phenylalanine methyl esters): wherein: the chiral centers of the compound of formula 1 are in the S configuration and the R configuration, respectively; and R _{1 is} selected from the group consisting of hydrogen atom, hydroxyl group and methoxy group ( One of Methoxy); R _{2 is} selected from the group consisting of a non-substituted phenyl group, a mono-substituted phenyl group, a di-substituted phenyl group, and a trisubstituted benzene. One of a tri-substituted phenyl group, a pyridinyl group, and a C ₄ -C ₆ cycloalkyl group; R _{3 is} selected from the group consisting of a non-substituted benzoyl group, a single Substituted benzoyl group, disubstituted benzoyl group (di-substit) One of uted benzoyl group) and a tri-substituted benzoyl group; and R _{4 is} selected from the group consisting of hydroxyl, C ₁ -C ₄ alkoxyl and glycine-glycin- One of nitrile).

【實施例十五】：如實施例十四所述的FPR1拮抗劑，其更包含該式一化合物在藥學上可接受的鹽類溶劑合物或其組合。 [Embodiment 15] The FPR1 antagonist according to embodiment 14, which further comprises the compound of the formula I in a pharmaceutically acceptable salt solvate or a combination thereof.

【實施例十六】：如實施例十四所述的FPR1拮抗劑，其中該嗜中性白血球發炎性障礙係選自於下列病症所組成之群組：肺損傷、慢性阻塞性肺臟疾病、急性呼吸窘迫症候群、氣喘、缺血性再灌流損傷、關節炎以及敗血病。 [16] The FPR1 antagonist according to the fourteenth aspect, wherein the neutrophil inflammatory disorder is selected from the group consisting of lung injury, chronic obstructive pulmonary disease, acute Respiratory distress syndrome, asthma, ischemic reperfusion injury, arthritis, and septicemia.

【實施例十七】：一種雙胜肽類衍生物，其結構如式一所示，其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R 構型；R₁選自氫(hydrogen atom)、羥基(hydroxyl group)的其中之一；R₂選自無取代苯基(non-substituted phenyl group)、單取代苯基(mono-substituted phenyl group)、雙取代苯基(di-substituted phenyl group)、三取代苯基(tri-substituted phenyl group)、吡啶(pyridinyl)的其中之一；R₃選自無取代苯醯基團(non-substituted benzoyl group)、單取代苯醯基團(mono-substituted benzoyl group)、雙取代苯醯基團(di-substituted benzoyl group)和三取代苯醯基團(tri-substituted benzoyl group)的其中之一；以及R₄選自C₁-C₄烷氧基(alkoxyl)和甘氨酸氰(glycin-nitrile)的其中之一。 [Example] seventeen: Double Chance one kind of peptide derivatives, a structure as shown in formula, wherein: the center of the palm of a double compound (chiral centers) and S configuration, respectively R configuration; _{R. 1} One selected from the group consisting of a hydrogen atom and a hydroxyl group; R _{2 is} selected from a non-substituted phenyl group, a mono-substituted phenyl group, and a disubstituted phenyl group. One of di-substituted phenyl group, tri-substituted phenyl group, pyridinyl; R _{3 is} selected from non-substituted benzoyl group, monosubstituted benzene One of a mono-substituted benzoyl group, a di-substituted benzoyl group, and a tri-substituted benzoyl group; and R _{4 is} selected from C ₁ One of -C ₄ alkoxyl and glycine-nitrile.

【實施例十八】：如實施例十七所述的雙胜肽類衍生物的用途，在於該雙胜肽類衍生物可抑制並拮抗一甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)。 [Embodiment 18] The use of the bis-peptide derivative as described in Example 17 is that the bis-peptide derivative inhibits and antagonizes a methicone peptide receptor type 1 (Formyl Peptide) Receptor 1, FPR1).

【實施例十九】：如實施例十八所述的用途，其中，該雙胜肽類衍生物抑制一類fMLP的衍生物N-甲醯甲硫氨醯亮氨醯苯丙氨酸螢光(N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein)FNLFNYK結合至該FPR1。 [Embodiment 19] The use according to the embodiment 18, wherein the bis-peptide derivative inhibits a derivative of a class of fMLP, N -methyl sulfoxime, leucine, phenylalanine, fluorescein ( N- formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein)FNLFNYK binds to the FPR1.

【實施例二十】：如實施例十八所述的用途，其中，該雙胜肽類衍生物抑制FPR1下游鈣離子(Calcium ion)、有絲***活化蛋白質激酶(mitogen-activated protein kinases,MAPKs)和蛋白質激酶B(protein kinase B,Akt)的至少其中之一的訊息傳遞。 [Embodiment 20] The use according to the embodiment 18, wherein the bis-peptide derivative inhibits calcium ion, mitogen-activated protein kinases (MAPKs), and Signal transfer of at least one of protein kinase B (Akt).

【實施例二十一】：如實施例十八所述的用途，其中該雙胜肽類衍生物競爭性抑制一FPR1活化物所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧化基的生成。 [Embodiment 21] The use according to the embodiment 18, wherein the bis-peptide derivative competitively inhibits release of human neutrophil elastase induced by an FPR1 activator and superoxide-based generate.

【實施例二十二】：如實施例二十一所述的用途，其中：該FPR1活化物係源自於一嗜中性白血球發炎性障礙；以及該嗜中性白血球發炎性障礙係至少一選自於下列病症所組成之群組：肺損傷、慢性阻塞性肺炎、急性呼吸窘迫症候群、氣喘、缺血性再灌流損傷、關節炎以及敗血病。 [Embodiment 22] The use of embodiment 21, wherein: the FPR1 activator is derived from a neutrophil inflammatory disorder; and the neutrophil inflammatory disorder is at least one It is selected from the group consisting of lung injury, chronic obstructive pneumonia, acute respiratory distress syndrome, asthma, ischemic reperfusion injury, arthritis, and septicemia.

【實施例二十三】：一種雙胜肽類衍生物，其結構如式一所示，其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；R₁選自氫；R₂選自無取代苯基；R₃選自無取代苯醯基團；以及R₄選自C₁烷氧基。 [Example 23]: a double peptide derivative, the structure of which is shown in Formula 1, wherein: the chiral centers of the compound of the formula 1 are the S configuration and the R configuration, respectively; _{1 is} selected from hydrogen; R _{2 is} selected from unsubstituted phenyl; R _{3 is} selected from unsubstituted benzoquinone groups; and R _{4 is} selected from C ₁ alkoxy.

【實施例二十四】：一種雙胜肽類衍生物，其結構如式一所示，其中：該式一化合物的雙對掌中心(chiral centers)分別為S構型與R構型；R₁選自氫；R₂選自C₆環烷基；R₃選自無取代苯醯基團；以及R₄選自C₁烷氧基。 [Embodiment 24]: a double peptide derivative, the structure of which is shown in Formula 1, wherein: the chiral centers of the compound of the formula 1 are the S configuration and the R configuration, respectively; _{1 is} selected from hydrogen; R _{2 is} selected from a C ₆ cycloalkyl group; R _{3 is} selected from an unsubstituted benzoquinone group; and R _{4 is} selected from a C ₁ alkoxy group.

綜上所述，本案以較簡單的合成方法，合成一系列雙胜肽類衍生物，而此雙胜肽類衍生物其兩胺基酸需分別為L(或S)與D(或R)構型，另在N端與C端皆有取代基。實驗發現此系列化合物在人類嗜中性白血球實驗模式下對於抑制由FPR1專一性刺激劑fMLF所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成皆有極佳的抑制作用。但對於由FPR2專一性刺激劑WKYMVm所誘導的人類嗜中性白血球彈性蛋白酶的釋放與超氧自由基的生成雖有抑制效果，但較FPR1專一性刺激劑所誘導的抑制活性明顯降低約3~20倍，顯示此系列新穎的雙胜肽類衍生物對於FPR1具有選擇性的拮抗作用。 In summary, this case synthesizes a series of double peptide derivatives by a simpler synthesis method, and the two amino acids of the double peptide derivatives need to be L (or S ) and D (or R ), respectively. The configuration has a substituent at both the N-terminus and the C-terminus. It was found that this series of compounds have excellent inhibitory effects on the release of human neutrophil elastase induced by the FPR1 specific stimulator fMLF and the production of superoxide radicals in the human neutrophil experimental mode. However, the release of human neutrophil elastase induced by the FPR2 specific stimulator WKYMVm and the formation of superoxide radicals were inhibited, but the inhibitory activity induced by the FPR1 specific stimulator was significantly reduced by about 3~. 20 times, showing that this series of novel double peptide derivatives have selective antagonism for FPR1.

若進一步地進行雙胜肽類衍生物的藥理機轉探討，其結果顯示化合物3(HCH6-1)在無細胞系統的實驗中，並無改變超氧自由基和彈性蛋白酶釋放的作用。除此之外，藉由非FPR1刺激劑活化之下，化合物3(HCH6-1)僅具些微的影響。有趣的是，化合物3(HCH6-1)具競爭性抑制FPR1下游的訊息傳遞路徑，包含鈣離子、MAPKs以及Akt。綜合實驗結果可以得知化合物3(HCH6-1)為一選擇性和競爭性地抑制FPR1誘使人類嗜中性白血球活化。符合此假設，HCH6-1具有抑制N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein(一種類fMLP的螢光衍生物)結合FPR的能力。於人類嗜中性白血球中，化合物3(HCH6-1)具競爭性的抑制FPR1的能力。 Further investigation of the pharmacokinetics of the bis- peptide derivatives revealed that the compound 3 (HCH6-1) did not change the effects of superoxide radicals and elastase release in the cell-free system experiment. In addition to this, compound 3 (HCH6-1) has only a slight effect by activation with non-FPR1 stimulator. Interestingly, Compound 3 (HCH6-1) competitively inhibits the signaling pathway downstream of FPR1, including calcium ions, MAPKs, and Akt. Based on the results of the comprehensive experiment, it was found that Compound 3 (HCH6-1) selectively and competitively inhibits FPR1 from inducing human neutrophil activation. Consistent with this hypothesis, HCH6-1 has the ability to inhibit the binding of N- formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein, a fluorescent derivative of fMLP-like, to FPR. Compound 3 (HCH6-1) is competitively capable of inhibiting FPR1 in human neutrophils.

另外，雙胜肽類衍生物在合成的過程中，可分別獲得N端具有(E)-N-benzoyl或(Z)-N-benzoyl取代基團的兩種異構物，上述化合物利用圓二色光譜儀分析結果顯示，具有(E)-N-benzoyl的雙胜肽類衍生物其在波長220nm的科頓效應(Cotton Effect)為負值，而具有(Z)-N-benzoyl的雙胜肽類衍生物其在波長220nm的Cotton effect則為正值。在活性評估上，具有(Z)-N-benzoyl的雙胜肽類衍生物則明顯高於具有(E)-N-benzoyl的雙胜肽類衍生物。 In addition, in the synthesis process, the two peptides have two isomers of ( E ) -N- benzoyl or ( Z ) -N- benzoyl substituent groups at the N- terminus, and the above compounds utilize round two The results of color spectrometer analysis showed that the double peptide derivative with ( E ) -N- benzoyl had a negative Cotton effect at a wavelength of 220 nm, while the double peptide with ( Z ) -N- benzoyl The derivative is positive at the Cotton effect at a wavelength of 220 nm. In the activity evaluation, the double peptide derivative with ( Z ) -N- benzoyl was significantly higher than the double peptide derivative with ( E ) -N- benzoyl.

本案雙胜肽衍生物則可抑制fMLF所誘導的嗜中性白血球彈性蛋白酶與超氧自由基的生成具有極佳的抑制作用。且經由進一步藥理活性評估結果顯示，上述雙胜肽類衍生物具競爭性抑制FPR1下游的訊息傳遞路徑，包含鈣離子、MAPKs以及Akt。表一之綜合實驗結果可以得知其中化合物3(HCH6-1)與24a為選擇性和競爭性地抑制FPR1誘使人類嗜中性白血球活化；同時化合物3(HCH6-1)具有抑制一種類fMLF的N-甲醯甲硫氨醯亮氨醯苯丙氨酸螢光衍生物(N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein)結合FPR的能力。因此，雙胜肽類衍生物可做為FPR1拮抗劑。 In this case, the double peptide derivative can inhibit the formation of neutrophil elastase and superoxide radical induced by fMLF. Furthermore, the results of further pharmacological activity evaluation showed that the above-mentioned double peptide derivatives competitively inhibit the signal transduction pathway downstream of FPR1, including calcium ions, MAPKs and Akt. The comprehensive experimental results in Table 1 show that Compound 3 (HCH6-1) and 24a selectively and competitively inhibit FPR1 to induce human neutrophil activation; while Compound 3 (HCH6-1) inhibits a class of fMLF The ability of the N- formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein to bind to FPR. Therefore, the double peptide derivative can be used as an FPR1 antagonist.

參考文獻 references

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4. Hwang TL, Hung CH, Hsu CY, Huang YT, Tsai YC, Hsieh PW., Design and synthesis of tryptophan containing dipeptide derivatives as formyl peptide receptor 1 antagonist. Org Biomol Chem. 2013 Jun 14;11(22):3742-55. 4. Hwang TL, Hung CH, Hsu CY, Huang YT, Tsai YC, Hsieh PW., Design and synthesis of tryptophan containing dipeptide derivatives as formyl peptide receptor 1 antagonist. Org Biomol Chem. 2013 Jun 14;11(22):3742 -55.

5. 黃吟婷(2013)。新雙胜肽HCH6-1藉由選擇性拮抗FPR1抑制發炎反應。長庚大學中醫系天然藥物碩士班碩士論文。台灣。 5. Huang Yuting (2013). The new double peptide HCH6-1 inhibits the inflammatory response by selectively antagonizing FPR1 . Master's thesis of the Master of Natural Medicines, Department of Traditional Chinese Medicine, Chang Gung University. Taiwan.

Claims

一種組合物之用途，係製備可治療甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)拮抗劑之組合物，其包括如式四所示的化合物N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine)衍生物：其中：其中：該式四化合物的雙對掌中心(chiral centers)分別為S構型與R構型；其中R1選自氫基(hydrogen atom)、羥基(hydroxyl group)、C₁-C₄烷基取代之羥基、C₁-C₄烷氧基(alkoxyl group)其中之一；R4選自氫基、羥基；含有C₁-C₄烷基取代之羥基；醯胺基團含有C₁-C₄烷基(alkyl)氰基之取代、或C₁-C₄烷基或C₁-C₄烷氧基之取代。 A composition for the preparation of a composition for treating a Formyl Peptide Receptor (FPR1) antagonist comprising a compound N- ( N -aralkyl) as shown in Formula IV Derivatives of N- ( N- aroyl-L-tryptophanyl)-D-phenylalanine: Wherein: wherein the chi-centers of the four compounds of the formula are S configuration and R configuration, respectively; wherein R1 is selected from a hydrogen atom, a hydroxyl group, a C ₁ -C ₄ alkane. a group substituted with a hydroxyl group, one of a C ₁ -C ₄ alkoxyl group; R 4 is selected from a hydrogen group, a hydroxyl group; a C ₁ -C ₄ alkyl group-substituted hydroxyl group; and a guanamine group containing a C ₁ -C group ₄ alkyl (alkyl) substituted by a cyano, or C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy groups substituted.

如請求項1所述組合物之用途，其更包含該式一化合物在藥學上可接受的鹽類或其組合。 The use of the composition of claim 1, which further comprises a pharmaceutically acceptable salt of the compound of the formula I or a combination thereof.

一種化合物之用途，係製備可治療甲醯化胜肽受體第1型(Formyl Peptide Receptor 1,FPR1)拮抗劑之化合物，其包括如式四所示的化合物N-(N-芳烷基-L-色氨醯基)-D-苯丙氨酸(N-(N-aroyl-L-tryptophanyl)-D-phenylalanine)衍生物：其中：其中：該式四化合物的雙對掌中心(chiral centers)分別為S構型與R構型；其中R1選自氫基(hydrogen atom)、羥基(hydroxyl group)、C₁-C₄烷基取代之羥基、C₁-C₄烷氧基(alkoxyl group)其中之一；R4選自氫基、羥基；含有C₁-C₄烷基取代之羥基；醯胺基團含有C₁-C₄烷基(alkyl)氰基之取代、或C₁-C₄烷基或C₁-C₄烷氧基之取代。 A compound for use in the preparation of a compound which treats a Formyl Peptide Receptor 1, FPR1 antagonist, comprising a compound N- ( N -aralkyl) as shown in Formula IV acyl L- tryptophyl) -D- phenylalanine (N - (N -aroyl-L -tryptophanyl) -D-phenylalanine) derivative: Wherein: wherein the chi-centers of the four compounds of the formula are S configuration and R configuration, respectively; wherein R1 is selected from a hydrogen atom, a hydroxyl group, a C ₁ -C ₄ alkane. a group substituted with a hydroxyl group, one of a C ₁ -C ₄ alkoxyl group; R 4 is selected from a hydrogen group, a hydroxyl group; a C ₁ -C ₄ alkyl group-substituted hydroxyl group; and a guanamine group containing a C ₁ -C group ₄ alkyl (alkyl) substituted by a cyano, or C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy groups substituted.