TWI428139B

TWI428139B - A novel glucagon-like peptide analogue, a composition and use thereof

Info

Publication number: TWI428139B
Application number: TW100141760A
Authority: TW
Original assignee: Betta Pharmaceuticals Co
Priority date: 2011-11-16
Filing date: 2011-11-16
Publication date: 2014-03-01
Also published as: TW201321015A

Description

一種新胰高血糖素樣肽類似物、組合物及其用途Novel glucagon-like peptide analog, composition and use thereof

本發明涉及一種新型胰高血糖素樣肽類似物及其組合物，用於調節哺乳動物體內胰島素的表達和治療糖尿病。特別地，這些多肽衍生物用於長效治療下述病症：糖尿病和其他與胰島素分泌多肽相關的疾病、與胰高血糖素水準相關的胃腸道功能和活性的疾病。本發明還涉及所述胰高血糖素樣肽類似物及其組合物用於上述病症的用途。The present invention relates to a novel glucagon-like peptide analog and a composition thereof for use in modulating insulin expression and treating diabetes in a mammal. In particular, these polypeptide derivatives are useful for the long-term treatment of diseases such as diabetes and other diseases associated with insulin secreting polypeptides, gastrointestinal function and activity associated with glucagon levels. The invention also relates to the use of said glucagon-like peptide analogs and compositions thereof for use in the above conditions.

胰島內分泌過程由一套複雜的控制機制調節。該機制不僅受血源性代謝物如葡萄糖、氨基酸和兒茶酚胺的影響，還受局部旁分泌的影響。主要的胰島激素、胰高血糖素、胰島素和生長抑制素可與特定的胰島細胞(A、B和D細胞)相互作用調節分泌反應。雖然胰島素分泌主要由血糖水準所決定，但是生長素抑制素也能抑制葡萄糖調節性血糖胰島素的分泌。除了胰島素分泌的內島旁分泌調節外，有事實證明存在腸內促胰島素因子。該促胰島素的概念起源於對以下現象的觀察：較相同數量的靜脈滴注能量(葡萄糖)，食物攝取或腸內葡萄糖吸收所獲得的能量能激起一個更大的刺激來促進胰島素的釋放(Elrick,H.,et al.,J.Clin. Endocrinol. Metab.1964,24:1076-1082；McIntyre,N.,et al.,.J. Clin. Endocrinol. Metab.,1965,25:1317-1324)。因此，假設當能量經胃腸道而非胃腸外途徑攝取時，口頭營養物質攝取所激發的腸源性信號代表了促使胰島素釋放增加的有效胰島素促分泌素(Dupre,J.,et al.,.Diabetes. 1966,15:555-559)。雖然一些神經傳遞質和胃腸激素也有類似腸促胰島素的活性，但是從免疫、拮抗劑和knockout的研究中所得到的大量證據表明，葡萄糖依賴性促胰島素多肽(Glucose-dependent insulinotropic polypeptide,GIP)和胰高血糖素樣肽-1(Glucagons-like peptide-1,GLP-1)才代表了對大多數營養物質刺激所引起的胰島素分泌起主導作用的多肽。研究表明，II型糖尿病患者在進餐所刺激的胰島素分泌釋放中表現出明顯降低，這一觀察結果引起了人們對以下研究的興趣：確定有缺陷的腸促胰島素釋放或腸促胰島素作用抗性是否與糖尿病患者β-細胞功能紊亂的病理生理學有關。The islet endocrine process is regulated by a complex set of control mechanisms. This mechanism is not only affected by blood-derived metabolites such as glucose, amino acids and catecholamines, but also by local paracrine. The major islet hormones, glucagon, insulin, and somatostatin interact with specific islet cells (A, B, and D cells) to regulate the secretory response. Although insulin secretion is mainly determined by blood glucose levels, auxin can also inhibit the secretion of glucose-regulated glycemic insulin. In addition to the endo-colonial regulation of insulin secretion, there is evidence of the presence of intestinal insulinotropic factors. The concept of insulinotropics stems from the observation that the same amount of intravenous infusion of energy (glucose), energy from food intake or intestinal glucose absorption can provoke a greater stimulus to promote insulin release ( Elrick, H., et al., J. Clin. Endocrinol. Metab. 1964, 24: 1076-1082; McIntyre, N., et al., J. Clin. Endocrinol. Metab., 1965, 25: 1317- 1324). Thus, assuming that when energy is taken up through the gastrointestinal tract rather than the parenteral route, the intestinal-derived signal stimulated by oral nutrient uptake represents an effective insulin secretagogue that promotes increased insulin release (Dupre, J., et al.,. Diabetes. 1966, 15: 555-559). Although some neurotransmitters and gastrointestinal hormones have similar incretin-like activities, a large body of evidence from studies of immunization, antagonists, and knockout suggests that Glucose-dependent insulinotropic polypeptide (GIP) and Glucagons-like peptide-1 (GLP-1) represents a polypeptide that plays a leading role in insulin secretion caused by most nutrient stimulation. Studies have shown that patients with type 2 diabetes show a significant reduction in insulin-stimulated release stimulated by meals, and this observation has raised interest in the following studies: determining whether defective incretin release or incretin resistance is resistant It is related to the pathophysiology of β-cell dysfunction in diabetic patients.

1987年，人類首次發現了胰高血糖素樣肽-1(GLP-1)，認為它是一種內分泌激素，是一種在食物攝取後由腸道分泌的多肽。GLP-1是由160個氨基酸前體蛋白(即前胰高血糖素原)經蛋白水解過程之後由腸L細胞分泌的。前胰高血糖素原經裂解後首先生成一種由37個氨基酸組成的肽GLP-1，即GLP-1(1-37)OH，該肽的活性很弱。隨後，再在7-位上裂解生成有生物活性的GLP-1(7-37)OH。在L-細胞的末端甘氨酸殘基被移除後，大約80%合成的GLP-1(7-37)OH在C-末端上被醯胺化。游離酸GLP-1(7-37)OH和醯胺化合物GLP-1(7-37)NH₂ 的生物學影響和代謝更新無法區分。In 1987, human glucagon-like peptide-1 (GLP-1) was first discovered as an endocrine hormone, a polypeptide secreted by the intestine after food intake. GLP-1 is secreted by intestinal L cells after proteolytic process by a 160 amino acid precursor protein (ie, proglucagon). After cleavage of proglucagon, a peptide GLP-1 consisting of 37 amino acids, GLP-1(1-37)OH, is first formed, and the activity of the peptide is very weak. Subsequently, it is cleaved at the 7-position to produce bioactive GLP-1(7-37)OH. After the glycine residue at the end of the L-cell was removed, approximately 80% of the synthesized GLP-1 (7-37) OH was amylated on the C-terminus. The biological effects and metabolic turnover of the free acid GLP-1 (7-37) OH and the guanamine compound GLP-1 (7-37) NH ₂ cannot be distinguished.

眾所周知，GLP-1能夠刺激引起細胞葡萄糖攝取的胰島素分泌，從而降低血糖水準(Mojsov,S.,et al.,J. Clin. Invest.,1987,79:616-619；Kreymann,B.,et al.,Lancet ii,1987,1300-1304；Orskov,C.,et al.,Endocrinology,1988,123:2009-2013)。急性腦室內注射GLP-1或GLP-1受體激動劑會產生短暫的食物攝取減少(Turton M.D.,et al.,Nature,1996,379:60-72)。然而，一些研究表明，更多長期的腦室內或胃腸外GLP-1受體激動劑的給藥與體重減輕有關(Meeran,K.,et al.,Endocrinology,1999,140:244-250；Davies,H.R.Jr.,Obes.Res.,1998,6:147-156；Szayna,M.,et al.,Endocrinology,2000,141:1936-1941；Larsen,P.J.,et al.,Diabetes,2001,50:2530-2539)。大量具有促胰島素作用的GLP-1類似物在本領域是已知的。例如，這些類似物包括GLP-1(7-36)、Gln⁹ -GLP-1(7-37)、D-Gln9-GLP-1(7-37)、乙醯基-Lys9-GLP-1(7-37)、Thr16-Lys18-GLP-1(7-37)和Lys18-GLP-1(7-37)。例如，GLP-1衍生物包括酸加成鹽、羧酸鹽、低級烷基酯和醯胺類化合物(WO91/11457、EP0733644及US5512549)。GLP-1 is known to stimulate insulin secretion that causes glucose uptake by cells, thereby lowering blood glucose levels (Mojsov, S., et al., J. Clin. Invest., 1987, 79: 616-619; Kreymann, B., et Al., Lancet ii, 1987, 1300-1304; Orskov, C., et al., Endocrinology, 1988, 123: 2009-2013). Acute intraventricular injection of GLP-1 or GLP-1 receptor agonists produces a transient reduction in food intake (Turton MD, et al., Nature, 1996, 379: 60-72). However, some studies have shown that administration of more long-term intraventricular or parenteral GLP-1 receptor agonists is associated with weight loss (Meeran, K., et al., Endocrinology, 1999, 140: 244-250; Davies , HR Jr., Obes. Res., 1998, 6: 147-156; Szayna, M., et al., Endocrinology, 2000, 141: 1936-1941; Larsen, PJ, et al., Diabetes, 2001, 50: 2530-2539). A large number of GLP-1 analogs with insulinotropic effects are known in the art. For example, these analogs include GLP-1 (7-36), Gln ⁹ -GLP-1 (7-37), D-Gln9-GLP-1 (7-37), acetyl-Lys9-GLP-1 ( 7-37), Thr16-Lys18-GLP-1 (7-37) and Lys18-GLP-1 (7-37). For example, GLP-1 derivatives include acid addition salts, carboxylates, lower alkyl esters, and guanamine compounds (WO 91/11457, EP 0733644, and US 5,512,549).

大多數在臨床前實驗中敍述的GLP-1作用在人體研究中也已經獲得了證實。在葡萄糖負載或食物攝取消化狀態下，對正常人體內注射GLP-1(7-36)NH₂ 會刺激胰島素分泌，明顯降低血糖水準(Orskov,C.,et al.,Diabetes,1993,42:658-661；Qualmann,C.,et al.,Acta.Diabetol.,1995,32:13-16)。Most of the effects of GLP-1 described in preclinical experiments have also been demonstrated in human studies. In the glucose-loaded or food-ingested state, injection of GLP-1(7-36)NH ₂ into normal humans stimulates insulin secretion and significantly lowers blood glucose levels (Orskov, C., et al., Diabetes, 1993, 42: 658-661; Qualmann, C., et al., Acta. Diabetol., 1995, 32: 13-16).

對於磺脲類藥物治療失敗的糖尿病患者而言，選用GLP-1類似肽來替代胰島素治療肯定會有良好的療效(Nauck,M.A.,et al.,Diabetes Care,1998,21:1925-1931)。GLP-1只在高血糖症的情況下刺激胰島素分泌。GLP-1的這個特性使其和胰島素相比起來更具有安全性，而且已經觀察到分泌胰島素的數量和高血糖症的程度是成比例的。另外，GLP-1治療法將導致胰腺釋放胰島素並使胰島素首先作用於肝臟。和皮下胰島素注射相比，這些結果可導致外周胰島素的低水準循環。GLP-1使胃腸道清空過程變慢，這更有利於讓營養物質在較長的時間內被吸收，從而降低餐後葡萄糖峰值。一些報導指出GLP-1能夠提高外周組織(如肌肉、肝臟和脂肪)中胰島素的敏感性。GLP-1還可作為潛在的食欲調節劑。For diabetic patients who have failed sulfonylurea therapy, the use of GLP-1-like peptides for insulin therapy will certainly have a good effect (Nauck, M.A., et al., Diabetes Care, 1998, 21:1925-1931). GLP-1 stimulates insulin secretion only in the case of hyperglycemia. This property of GLP-1 makes it safer than insulin, and it has been observed that the amount of insulin secreted is proportional to the degree of hyperglycemia. In addition, GLP-1 treatment will result in the release of insulin from the pancreas and the first action of insulin on the liver. These results can result in a low level of circulating insulin compared to subcutaneous insulin injections. GLP-1 slows the gastrointestinal emptying process, which is more conducive to allowing nutrients to be absorbed over a longer period of time, thereby reducing postprandial glucose peaks. Some reports indicate that GLP-1 can increase the sensitivity of insulin in peripheral tissues such as muscle, liver and fat. GLP-1 also acts as a potential appetite regulator.

如果考慮應用於I型糖尿病患者中，GLP-1及其類似物在治療方面的潛能會更加突出。大量研究證明了天然GLP-1在治療胰島素依賴性糖尿病(IDDM)中的有效性。與非胰島素依賴性糖尿病(NIDDM)患者類似，GLP-1通過其穩定胰高血糖素的特性，能有效地緩解餐前高血糖症。其他研究還表明，很可能通過延長胃的清空，GLP-1還能減少IDDM患者餐後血糖偏移。這些觀察結果表明，GLP-1可用於治療IDDM和NIDDM。If considered for use in patients with type 1 diabetes, the potential for treatment of GLP-1 and its analogs will be more prominent. Numerous studies have demonstrated the effectiveness of native GLP-1 in the treatment of insulin-dependent diabetes mellitus (IDDM). Similar to patients with non-insulin-dependent diabetes mellitus (NIDDM), GLP-1 is effective in alleviating pre-prandial hyperglycemia by stabilizing glucagon properties. Other studies have also shown that GLP-1 can also reduce postprandial glucose excursion in IDDM patients by prolonging gastric emptying. These observations indicate that GLP-1 can be used to treat IDDM and NIDDM.

然而，天然GLP-1分子的生物半衰期受二肽酶IV(DPPIV)活性的影響，其生物半衰期非常短。例如，GLP-1(7-37)OH的生物學半衰期僅為3-5分鐘(US5118666)。只有通過持續滴注GLP-1才能持續降低血糖濃度，這就像研究中所證明的那樣，24小時以上的靜脈滴注給藥才能控制血糖濃度(Larsen,J.,et al.,Diabetes Care,2001,24:1416-1421)。酶DPP IV是一種絲氨酸蛋白酶，可優先將倒數第二個位置上為脯氨酸(Xaa-Pro-)或丙氨酸(Xaa-Ala-)的NH₂ -末端肽水解下來(Mentlein,R.,Regul. Pept.,1999,85:9-25)，DPP IV酶已經表現出能夠在體外迅速代謝GLP-1的能力。因此，對DPP IV有抗性的長效GLP-1類似肽在治療糖尿病方面具有巨大的治療潛能。However, the biological half-life of native GLP-1 molecules is affected by dipeptidase IV (DPPIV) activity, which has a very short biological half-life. For example, the biological half-life of GLP-1(7-37)OH is only 3-5 minutes (US5118666). Only by continuous instillation of GLP-1 can the blood glucose concentration be continuously reduced, as demonstrated by the study, which can control blood glucose levels by intravenous infusion over 24 hours (Larsen, J., et al., Diabetes Care, 2001, 24: 1416-1421). The enzyme DPP IV is a serine protease which preferentially hydrolyzes the NH ₂ -terminal peptide of the proline (Xaa-Pro-) or alanine (Xaa-Ala-) in the penultimate position (Mentlein, R. , Regul. Pept., 1999, 85: 9-25), DPP IV enzymes have demonstrated the ability to rapidly metabolize GLP-1 in vitro. Therefore, long-acting GLP-1-like peptides that are resistant to DPP IV have enormous therapeutic potential in the treatment of diabetes.

本發明的目的是提供一種新的並能發揮更持久作用的GLP-1類似物，與天然GLP-1相比，該GLP-1類似物對酶DPP IV的水解作用具有完全的抗性。It is an object of the present invention to provide a novel GLP-1 analog which is more durable and which is completely resistant to the hydrolysis of the enzyme DPP IV compared to native GLP-1.

本發明包括至少一種具有下述式(I)的化合物或其藥學上可接受的鹽，The present invention includes at least one compound having the following formula (I) or a pharmaceutically acceptable salt thereof,

Xaa₇ -Q-Gly-Thr-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Tyr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B　------------------(I)Xaa ₇ -Q-Gly-Thr-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Tyr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe- Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B ------------------(I)

其中，Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Wherein, Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, and high An amino acid of histidine, α-fluoromethyl-histidine or α-methyl-histidine;

Q選自下述連接子(II)、(III)、(IV)：Q is selected from the following linkers (II), (III), (IV):

式中，R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₃ 為H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z選自N、O或S時，W不存在；當Z是C時，W選自H或F；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸或γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸，B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。Wherein R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ - C ₆ ) alkoxy; R ₃ is H, (C ₁ -C ₆ )alkyl, or form a 5-8 membered ring with R ₁ or R ₂ ; X is selected from H, F, OH, CF ₃ or O; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is selected from N, O or S, W is absent; when Z is When C, W is selected from H or F; Xaa ₁₄ is a natural or non-natural amino acid selected from serine or histidine, wherein one or more carbon atoms of the amino acid may also be arbitrarily one or more Alkyl substitution; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine, wherein one or more carbon atoms of the amino acid may also be arbitrarily one or more Alkyl substitution; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine, wherein one or more carbons of said amino acid atoms may be optionally substituted with one or more alkyl groups; Xaa ₂₂ and Xaa ₂₃ are natural or unnatural, selected Amino acid, amino isobutyric acid or glutamic acid, wherein one or more carbon atoms of said amino acids may also be optionally substituted by one or more alkyl _{_{groups; Xaa 26, Xaa 27, Xaa}} 34, Xaa 35 And Xaa ₃₆ are natural or non-natural, an amino acid selected from the group consisting of glycine, lysine, arginine, leucine, aspartame or aminoisobutyric acid, wherein one or more carbon atoms of the amino acid It may also be optionally substituted by one or more alkyl groups; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid or HOOC (CH _{2 )} _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine or γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids, and B is selected from glycine, glycinamine or a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

本發明進一步提供了上述技術方案的優選技術方案：式(I)中，Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The present invention further provides a preferred technical solution of the above technical solution: in the formula (I), Xaa ₂₆ is natural or non-natural, selected from glycine, lysine, arginine, leucine and aspartame or not A-aminoisobutyric acid linked to TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

上述肽片段為半胱氨酸-絲氨酸-甘氨酸、半胱氨酸-丙氨酸或連接著半胱氨酸的甲氧基聚乙二醇馬來醯亞胺。The above peptide fragment is cysteine-serine-glycine, cysteine-alanine or methoxypolyethylene glycol maleimide linked to cysteine.

本發明還提供了一種式(V)所示的GLP-1類似物、其組合物或其藥學上可接受的鹽，The present invention also provides a GLP-1 analogue represented by the formula (V), a composition thereof, or a pharmaceutically acceptable salt thereof,

在式(V)中，R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R3選自H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；X選自H、F、OH、CF₃ 或O；D是Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸或γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。In formula (V), R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ ) alkoxy; R 3 is selected from H, (C ₁ -C ₆ )alkyl, or forms a 5-8 membered ring with R ₁ or R ₂ ; X is selected from H, F, OH, CF ₃ or O; D is Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile -Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine , amino acid of 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine; Xaa ₁₄ is natural or unnatural An amino acid selected from the group consisting of serine or histidine, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; Xaa ₁₆ is natural or non-natural, selected from the group consisting of hydrazine ammonia An amino acid of acid, lysine or leucine, wherein one or more carbon atoms of said amino acid may also be optionally substituted by one or more alkyl groups; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ Natural or unnatural, selected from serine, arginine or lysine amino acid, wherein one or more carbon atoms of said amino acids may also be optionally substituted by one or more alkyl groups; Xaa ₂₂ and Xaa ₂₃ Is a natural or non-natural amino acid selected from the group consisting of glycine, aminoisobutyric acid or glutamic acid, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural amino acids selected from the group consisting of glycine, lysine, arginine, leucine, aspartame or aminoisobutyric acid, wherein One or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ-glutamic acid, β-alanine, γ- Aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, ₁₆ , 17 , 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine or γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids; B is selected from glycine, glycosamine or a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

本發明還提供了上述技術方案的優選技術方案：上述式(V)中，D所含有的Xaa₂₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The present invention also provides a preferred technical solution of the above technical solution: in the above formula (V), the Xaa ₂₆ contained in D is natural or non-natural, and is selected from the group consisting of glycine, lysine, arginine, leucine and day. Winter oxime or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

上述式(V)中，R₁ 、R₂ 和R₃ 均選自H或CH₃ ，或者R₁ 是CH₃ ，R₂ 和R₃ 均是H；或者R₁ 和R₃ 均是H，R₂ 是CH₃ ；或者R₃ 與R₁ 形成5~8元環，R₂ 是H；或者R₃ 與R₂ 形成5~8元環，R₁ 是H。In the above formula (V), R ₁ , R ₂ and R ₃ are each selected from H or CH ₃ , or R ₁ is CH ₃ , R ₂ and R ₃ are both H; or both R ₁ and R ₃ are H, R ₂ is CH ₃ ; or R ₃ and R ₁ form a 5-8-membered ring, R ₂ is H; or R ₃ and R ₂ form a 5-8-membered ring, and R ₁ is H.

本發明還進一步提供了一種式(VI)所示的GLP-1類似物、其組合物或其藥學上可接受的鹽，The present invention still further provides a GLP-1 analogue represented by the formula (VI), a composition thereof, or a pharmaceutically acceptable salt thereof,

R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₃ 選自H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；Y選自H、OH、F或(C₁ -C₆ )烷基；D是Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸或γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ ) Alkoxy; R _{3 is} selected from H, (C ₁ -C ₆ )alkyl, or forms a 5-8 membered ring with R ₁ or R ₂ ; Y is selected from H, OH, F or (C ₁ -C ₆ ) Alkyl; D is Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile- Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, Amino acid of 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine; Xaa ₁₄ is natural or unnatural An amino acid selected from the group consisting of serine or histidine, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; Xaa ₁₆ is natural or non-natural, selected from the group consisting of valine An amino acid of lysine or leucine, wherein one or more carbon atoms of said amino acid may also be optionally substituted by one or more alkyl groups; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ Is natural Or non-natural, an amino acid selected from the group consisting of serine, arginine or lysine, wherein one or more carbon atoms of said amino acid may also be optionally substituted by one or more alkyl groups; Xaa ₂₂ and Xaa ₂₃ are A natural or non-natural amino acid selected from the group consisting of glycine, aminoisobutyric acid or glutamic acid, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural amino acids selected from the group consisting of glycine, lysine, arginine, leucine, aspartame or aminoisobutyric acid, wherein the amino acid One or more carbon atoms may also be optionally substituted by one or more alkyl groups; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ-glutamic acid, β-alanine, γ-amino Butyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, ₁₆ , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine or γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids; B is selected from glycine, glycosamine or a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

本發明還提供了上述技術方案的優選技術方案：上述式(VI)中，D所含有的Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The present invention also provides a preferred technical solution of the above technical solution: in the above formula (VI), X contains Xaa _{26 which} is natural or non-natural, and is selected from the group consisting of glycine, lysine, arginine, leucine and day. Winter oxime or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

上述式(VI)中，R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基；R₃ 是H；Y選自H或(C₁ -C₆ )烷基。In the above formula (VI), R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl; R ₃ is H; Y is selected from H or ( C ₁ -C ₆ )alkyl.

本發明又進一步提供了一種式(VII)所示的GLP-1類似物、其組合物或其藥學上可接受的鹽，The present invention still further provides a GLP-1 analogue represented by the formula (VII), a composition thereof, or a pharmaceutically acceptable salt thereof,

在式(VII)中，R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₃ 選自H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；Y選自H、OH、F或(C₁ -C₆ )烷基；W選自H或F；.D是Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自。In formula (VII), R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{3 is} selected from H, (C ₁ -C ₆ )alkyl, or forms a 5-8 membered ring with R ₁ or R ₂ ; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; W is selected from H or F; D is Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa ₇ is natural or non-natural, selected from L-histamine Acid, D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl An amino acid of histidine; Xaa ₁₄ is a natural or non-natural amino acid selected from the group consisting of serine or histidine, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups. Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups. ; or Xaa ₁₆ is connected TU lysine; Xaa ₁₈ is natural or unnatural, selected from serine, arginine or lysine amino acid, wherein said one or more amino acids of carbon atoms can optionally be substituted with one or more Alkyl substitution; Xaa ₂₂ and Xaa ₂₃ are natural or non-natural, an amino acid selected from the group consisting of glycine, aminoisobutyric acid or glutamic acid, wherein one or more carbon atoms of the amino acid may also be arbitrarily one or Multiple alkyl substitutions; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural, selected from glycine, lysine, arginine, leucine, aspartame or amino An amino acid of butyric acid, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ-glutamic acid , β-alanine, γ-aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from .

式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸的氨基酸，或者T選自時存在，該式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Only T is an amino acid selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid, or T is selected from T In the formula, k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, and 9. Or 10; U is selected from C ₈ -C ₂₀ fatty acids; B is selected from glycine, glycosamine or a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

本發明還提供了上述技術方案的優選技術方案：上述式(VII)中，D所含有的Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The present invention also provides a preferred technical solution of the above technical solution: in the above formula (VII), the Xaa ₂₆ contained in D is natural or non-natural, and is selected from the group consisting of glycine, lysine, arginine, leucine and day. Winter oxime or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

上述式(VII)中，R₃ 是H，或與R₁ 或R₂ 形成5-8元環；Y是H或F。In the above formula (VII), R ₃ is H or forms a 5-8 membered ring with R ₁ or R ₂ ; Y is H or F.

本發明還提供了一種式(VIII)所示的GLP-1類似物、其組合物或其藥學上可接受的鹽，The present invention also provides a GLP-1 analogue represented by the formula (VIII), a composition thereof, or a pharmaceutically acceptable salt thereof,

其中，Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Q選自下述連接子(II)、(III)、(IV)：Wherein, Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, and high An amino acid of histidine, α-fluoromethyl-histidine or α-methyl-histidine; Q is selected from the following linkers (II), (III), (IV):

式中，R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₃ 選自H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z是N、O或S時，W不存在；當Z是C時，W選自H或F；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸，B選自甘氨酸或甘氨醯胺。Wherein R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ - C ₆ ) alkoxy; R _{3 is} selected from H, (C ₁ -C ₆ )alkyl, or forms a 5-8 membered ring with R ₁ or R ₂ ; X is selected from H, F, OH, CF ₃ or O Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is N, O or S, W is absent; when Z is When C, W is selected from H or F; Xaa ₁₄ is a natural or non-natural amino acid selected from serine or histidine, wherein one or more carbon atoms of the amino acid may also be arbitrarily one or more Alkyl substitution; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine, wherein one or more carbon atoms of the amino acid may also be arbitrarily one or more Alkyl substituted; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine, wherein one or more carbon atoms of the amino acid may also be optionally substituted by one or more alkyl groups. substituted; Xaa ₂₂ and Xaa ₂₃ are natural or unnatural, selected from glycine, aminoisobutyric acid or glutamic acid Acid, wherein one or more carbon atoms of said amino acids may also be optionally substituted by one or more alkyl _{_{groups; Xaa 27, Xaa 34, Xaa}} 35 and Xaa ₃₆ are natural or unnatural, selected from glycine, An amino acid of lysine, arginine, leucine, aspartame or aminoisobutyric acid, wherein one or more carbon atoms of said amino acid may also be optionally substituted by one or more alkyl groups; Selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids, and B is selected from glycine or glycine.

本發明還提供了一種式(IX)所示的GLP-1類似物、其組合物或其藥學上可接受的鹽，The present invention also provides a GLP-1 analogue represented by the formula (IX), a composition thereof, or a pharmaceutically acceptable salt thereof,

Xaa₇ -Q-Gly-Thr-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Tyr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -Gly-Xaaⁿ -Xaaⁿ⁺¹ -Xaaⁿ⁺² -Xaaⁿ⁺³ -Xaaⁿ⁺⁴ -Cys^(PEG) -Xaa^m -Xaa^m+1 -Xaa^m+2 -Xaa^m+3 -Xaa^m+4 IXXaa ₇ -Q-Gly-Thr-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Tyr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe- Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -Gly-Xaa ⁿ -Xaa ⁿ⁺¹ -Xaa ⁿ⁺² -Xaa ⁿ⁺³ -Xaa ⁿ⁺⁴ -Cys ^(PEG) -Xaa ^m -Xaa ^m+1 -Xaa ^m+2 -Xaa ^m+3 -Xaa ^m+4 IX

式中：Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Q選自下述連接子(II)、(III)、(IV)：Wherein Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, An amino acid of homohistidine, α-fluoromethyl-histidine or α-methyl-histidine; Q is selected from the following linkers (II), (III), (IV):

式中，R₁ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₂ 選自H、(C₁ -C₆ )烷基或(C₁ -C₆ )烷氧基；R₃ 為H、(C₁ -C₆ )烷基，或與R₁ 或R₂ 形成5-8元環；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z是N、O或S時，W不存在；當Z是C時，W選自H或F；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₂ 和Xaa₂₃ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸，其中，所述氨基酸的一個或多個碳原子也可以任意地被一個或多個烷基取代；或者Xaa₂₆ 是連接著T-U的賴氨酸；Xaaⁿ 、Xaaⁿ⁺¹ 、Xaaⁿ⁺² 、Xaaⁿ⁺³ 、Xaaⁿ⁺⁴ 、Xaa^m 、Xaa^m+1 、Xaa^m+2 、Xaa^m+3 及Xaa^m+4 一起組成1、2、3或4個天然的或非天然的氨基酸。也就是說，Xaaⁿ 、Xaaⁿ⁺¹ 、Xaaⁿ⁺² 、Xaaⁿ⁺³ 、Xaaⁿ⁺⁴ 、Xaa^m 、Xaa^m+1 、Xaa^m+2 、Xaa^m+3 和Xaa^m+4 與連接著甲氧基聚乙二醇馬來醯亞胺的半胱氨酸形成2~5個氨基酸的片段。Wherein R _{1 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R _{2 is} selected from H, (C ₁ -C ₆ )alkyl or (C ₁ - C ₆ ) alkoxy; R ₃ is H, (C ₁ -C ₆ )alkyl, or form a 5-8 membered ring with R ₁ or R ₂ ; X is selected from H, F, OH, CF ₃ or O; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is N, O or S, W is absent; when Z is C When W is selected from H or F; Xaa ₁₄ is a natural or non-natural amino acid selected from serine or histidine, wherein one or more carbon atoms of the amino acid may also be optionally substituted with one or more alkane Substituent; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine, wherein one or more carbon atoms of the amino acid may also be optionally substituted with one or more alkane Substituent; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine, wherein one or more carbon atoms of the amino acid may optionally be substituted with one or more alkyl groups; Xaa ₂₂ and Xaa ₂₃ are natural or unnatural, selected Gan Amino acids, amino isobutyric acid or glutamic acid, wherein one or more carbon atoms of the amino acid may optionally be substituted with one or more alkyl _{_{groups; Xaa 26, Xaa 27, Xaa}} 34, Xaa 35 , and Xaa ₃₆ is a natural or non-natural amino acid selected from the group consisting of glycine, lysine, arginine, leucine, aspartame or aminoisobutyric acid, wherein one or more carbon atoms of the amino acid are also It may be optionally substituted by one or more alkyl groups; or Xaa ₂₆ is a lysine linked to TU; Xaa ⁿ , Xaa ⁿ⁺¹ , Xaa ⁿ⁺² , Xaa ⁿ⁺³ , Xaa ⁿ⁺⁴ , Xaa ^m , Xaa ^m+1 , Xaa ^m+2 , Xaa ^m+3 and Xaa ^m+4 together constitute 1, 2, 3 or 4 natural or non-natural amino acids. That is, Xaa ⁿ , Xaa ⁿ⁺¹ , Xaa ⁿ⁺² , Xaa ⁿ⁺³ , Xaa ⁿ⁺⁴ , Xaa ^m , Xaa ^m+1 , Xaa ^m+2 , Xaa ^m+3 and Xaa ^m+4 The cysteine attached to the methoxypolyethylene glycol maleimide forms a fragment of 2 to 5 amino acids.

為幫助讀者理解本發明所涉及的化合物，本發明還提供選自如下的化合物：To assist the reader in understanding the compounds of the present invention, the present invention also provides a compound selected from the group consisting of:

‧[Q-連接子-d8,谷氨酸22]GLP-1-(7-37)-肽；‧[Q-linker-d8, glutamic acid 22] GLP-1-(7-37)-peptide;

‧[Q-連接子-a8-9,谷氨酸22]GLP-1-(7-37)-肽；‧[Q-linker-a8-9, glutamic acid 22] GLP-1-(7-37)-peptide;

‧[Q-連接子-b8-9,谷氨酸22]GLP-1-(7-37)-肽；‧[Q-linker-b8-9, glutamic acid 22] GLP-1-(7-37)-peptide;

‧[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽；‧[Q-linker-c8, glutamic acid 22] GLP-1-(7-37)-peptide;

‧[Q-連接子-e8-9,谷氨酸22]GLP-1-(7-37)-肽；‧[Q-linker-e8-9, glutamic acid 22] GLP-1-(7-37)-peptide;

‧[Q-連接子-f8-9,精氨酸34]GLP-1-(7-37)-肽；‧[Q-linker-f8-9, arginine 34] GLP-1-(7-37)-peptide;

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-c8, arginine 34]GLP-1-(7-37)- Peptide

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34]GLP-1-(7-37)- Peptide

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-_e 8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker- _e 8,arginine 34]GLP-1-(7-37) -peptide;

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-f8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-f8, arginine 34]GLP-1-(7-37)- Peptide

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-a8-9,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-a8-9,arginine 34]GLP-1-(7-37 )-peptide;

‧N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-b8-9,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-b8-9,arginine 34]GLP-1-(7-37 )-peptide;

‧N-ε²⁶ -[(N^ε -ω-羧基十七醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[(N ^ε -ω-carboxyheptadecanyl)]-[Q-linker-c8, arginine 34]GLP-1-(7-37)-peptide;

‧N-ε²⁶ -[(N^ε -ω-羧基十九醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；‧N-ε ²⁶ -[(N ^ε -ω-carboxynonanoyl)]-[Q-linker-c8, arginine 34] GLP-1-(7-37)-peptide;

‧[Q-連接子-d8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；‧ [Q-linker-d8] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;

‧[Q-連接子-c8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；‧ [Q-linker-c8] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;

‧[Q-連接子-a8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；‧ [Q-linker-a8-9] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;

‧[Q-連接子-b8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；‧ [Q-linker-b8-9] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;

‧[Q-連接子-e8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；‧ [Q-linker-e8-9] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;

‧[Q-連接子-f8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ 。‧[Q-linker-f8-9]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ .

本發明還提供了一種藥物組合物，包含有上述任一化合物及一種藥學上可接受的輔料。The invention also provides a pharmaceutical composition comprising any of the compounds described above and a pharmaceutically acceptable adjuvant.

本發明還提供了上述藥物組合物的優選技術方案：該藥物組合物適用於胃腸用藥。The present invention also provides a preferred technical solution of the above pharmaceutical composition: the pharmaceutical composition is suitable for gastrointestinal administration.

本發明還提供了上述任一化合物或藥物組合物在製備藥物中的應用。The invention also provides the use of any of the above compounds or pharmaceutical compositions for the preparation of a medicament.

本發明提供的任一化合物或藥物組合物在製備藥物中的優選技術方案：在製備治療或預防高血糖症、II型糖尿病、糖耐量受損、I型糖尿病、肥胖症、高血壓、X綜合征、血脂異常、認知障礙、動脈硬化、心肌梗塞、冠狀動脈心臟病和其他心血管疾病、中風、炎性腸道綜合征、消化不良和胃潰瘍藥物中的應用。A preferred technical solution for the preparation of a medicament by any of the compounds or pharmaceutical compositions provided by the present invention: in the preparation of a treatment or prevention of hyperglycemia, type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, X synthesis Applications in signs, dyslipidemia, cognitive impairment, arteriosclerosis, myocardial infarction, coronary heart disease and other cardiovascular diseases, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer medications.

在製備治療II型糖尿病的藥效延遲或預防惡化的藥物中的應用。The use in the preparation of a medicament for treating the delayed or preventable deterioration of the type 2 diabetes.

在製備用於減少食物攝入量、減少β細胞凋亡、增加胰島β細胞功能、增加β-細胞團和/或恢復葡萄糖對β-細胞的敏感性的藥物中的應用。Use in the preparation of a medicament for reducing food intake, reducing beta cell apoptosis, increasing islet beta cell function, increasing beta-cell mass and/or restoring glucose sensitivity to beta-cells.

本發明的核心是：用肽鍵模擬連接子替換GLP-1氨基末端的Ala⁸ 的氨基鍵，該氨基鍵是DPP-IV酶識別位點。所述肽鍵模擬連接子在藥物探索發現中是一種經典的方法，是通過模擬天然肽鍵且其保持了與生物靶點作用的能力並能產生相同的生物效應(Curr. Chem. Bio. 2008,12: 292-296)。基於相同的原理，所述肽鍵模擬連接子修飾的GLP-1類似物可能保持與天然GLP-1相同的生物活性，且具有長效促胰島素一樣持續作用。The core of the present invention is to replace the amino bond of Ala ⁸ at the amino terminus of GLP-1 with a peptide bond mimicking linker, which is a DPP-IV enzyme recognition site. The peptide bond mimic linker is a classical approach in drug discovery discovery by mimicking native peptide bonds and retaining the ability to interact with biological targets and produce the same biological effects (Curr. Chem. Bio. 2008) , 12: 292-296). Based on the same principle, the peptide-linked mock-modified GLP-1 analog may retain the same biological activity as native GLP-1 and have a sustained action like long-acting insulinotropic action.

本發明提供的化合物可能包含一個或多個不對稱中心，如式(I)的A-連接子。所述化合物可能具有一個或多個立體異構體。這些化合物可以是任意的活性形式，如外消旋體，富含對映異構體的立體異構體混合物。其中需要的單一對映異構體，如任意的活性形式，可以通過已知手段(如不對稱合成)採用光學活性起始物為原料合成，或外消旋拆分法獲得。外消旋拆分法可採用常規的方法完成。例如，在拆分試劑中結晶，在目標異構體分離後用對映純或富對映的拆分試劑中衍生化；或採用色譜法，如手性HPLC色譜柱。The compounds provided herein may contain one or more asymmetric centers, such as the A-linker of formula (I). The compound may have one or more stereoisomers. These compounds may be in any active form, such as a racemate, a mixture of stereoisomers enriched in enantiomers. The single enantiomers required therein, such as any of the active forms, can be synthesized by known means (e.g., asymmetric synthesis) using optically active starting materials, or by racemic resolution. The racemic resolution method can be carried out by a conventional method. For example, crystallization in a resolving reagent, derivatization with an enantiomerically or enantiomerically resolving reagent after separation of the target isomer; or chromatography, such as a chiral HPLC column.

本文中術語“多肽”和“肽”是指含有至少5個氨基酸組分通過肽鍵連接形成的化合物。所述氨基酸組分可選自被基因密碼編碼的氨基酸，不被基因密碼編碼的天然氨基酸以及合成氨基酸。所述不被基因密碼編碼的天然氨基酸可以是例如v-羧基谷氨酸酯、鳥氨酸、磷酸絲氨酸、D-丙氨酸和D-谷氨酸。合成氨基酸包括化學合成的氨基酸，例如基因密碼編碼的D-異構體，如D-丙氨酸和D-亮氨酸、α-氨基異丁酸(α-aminoisobutyric acid,Aib)、α-氨基丁酸(α-aminobutyric acid,Abu)、四丁基甘氨酸(tert-butylglycine,Tie)、β-丙氨酸、3-氨基甲基苯甲氨酸、鄰氨基苯甲酸。The terms "polypeptide" and "peptide" as used herein mean a compound containing at least 5 amino acid components joined by peptide bonds. The amino acid component may be selected from the group consisting of an amino acid encoded by a gene code, a natural amino acid not encoded by a gene code, and a synthetic amino acid. The natural amino acid not encoded by the genetic code may be, for example, v-carboxyglutamate, ornithine, phosphoserine, D-alanine, and D-glutamic acid. Synthetic amino acids include chemically synthesized amino acids, such as D-isomers encoded by the gene code, such as D-alanine and D-leucine, α-aminoisobutyric acid (Aib), α-amino --aminobutyric acid (Abu), tetrabutylglycine (Tie), β-alanine, 3-aminomethylphenyl methionine, anthranilic acid.

所述22種天然氨基酸是指：丙氨酸、精氨酸、天冬醯胺、天門冬氨酸、半胱氨酸、胱氨酸、谷氨醯胺、谷氨酸、甘氨酸、組氨酸、羥脯氨酸、異亮氨酸、亮氨酸、賴氨酸、蛋氨酸、苯丙氨酸、脯氨酸、絲氨酸、蘇氨酸、色氨酸、酪氨酸、纈氨酸。The 22 natural amino acids are: alanine, arginine, aspartame, aspartic acid, cysteine, cystine, glutamine, glutamic acid, glycine, histidine , hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, valine, serine, threonine, tryptophan, tyrosine, valine.

因此，非天然氨基酸作為一個成分並通過肽鍵與肽結合可以納入肽範疇，而不是天然氨基酸。具體實例如γ-羧基谷氨酸，鳥氨酸，磷酸絲氨酸，D-氨基酸如D-丙氨酸和D-谷氨酸。合成的非天然氨基酸包括化學合成製備的氨基酸，例如基因密碼編碼的氨基酸的D-異構體，例如D-丙氨酸和D-亮氨酸、α-氨基異丁酸(α-aminoisobutyric acid,Aib)、α-氨基丁酸(α-aminobutyric acid,Abu)、四丁基甘氨酸(tert-butylglycine,Tie)、3-氨基甲基苯甲酸、氨基苯甲酸、去氨基組氨酸，氨基酸β類似物如β-丙氨酸等。D-組氨酸、去氨基組氨酸、2-氨基-組氨酸、β-羥基-脯氨酸、高組氨酸、N^α -乙醯基-組氨酸、α-氟甲基-組氨酸、α-甲基-組氨酸、3-吡啶基丙氨酸、2-吡啶基丙氨酸或4-吡啶基丙氨酸、(1-氨基-環丙基)羧酸、(1-氨基環丁基)羧酸、(1-氨基環戊基)羧酸、(1-氨基環己基)羧基、(1-氨基環庚基)羧酸或(1-氨基環辛基)羧酸；關於GLP氨基酸序列已有報導(Lopez,L. C. et al.,Proc. Nat’l,Acad.Sci.,USA80 ,5485-5489,1983;Bell,G.I.,et al.,Nature302 :716-718(1983);Heinrich,G.,et al,Endocrinol,115 :2176-2181(1984)。前胰高血糖素原mRNA的結構及其相應的氨基酸序列是為公眾已知的。已有研究對該前體基因產物和前胰高血糖素水解成胰高血糖素和促胰島素釋放肽過程進行了表徵。本文中所述GLP-1(1-37)的標記是指含有所有從1位(N-端)到37位(C-端)氨基酸的GLP-1多肽。類似的，本文中所述GLP-1(7-37)的標記是指含有所有從7位(N-端)到37位(C-端)氨基酸的GLP-1多肽。類似的，本文中所述GLP-1(7-36)的標記是指含有所有從7位(N-端)到36位(C-端)氨基酸的GLP-1多肽。Therefore, an unnatural amino acid as a component and bound to a peptide by a peptide bond can be incorporated into the peptide category instead of the natural amino acid. Specific examples are γ-carboxyglutamic acid, ornithine, phosphoserine, D-amino acids such as D-alanine and D-glutamic acid. Synthetic non-natural amino acids include chemically synthesized amino acids, such as D-isomers of amino acids encoded by the gene code, such as D-alanine and D-leucine, alpha-aminoisobutyric acid (α-aminoisobutyric acid, Aib), α-aminobutyric acid (Abu), tetrabutylglycine (Tie), 3-aminomethylbenzoic acid, aminobenzoic acid, deaminohistidine, amino acid β Such as β-alanine and the like. D-histidine, deaminohistidine, 2-amino-histidine, β-hydroxy-valine, homohistidine, N ^α -acetyl-histidine, α-fluoromethyl- Histidine, α-methyl-histidine, 3-pyridyl alanine, 2-pyridyl alanine or 4-pyridyl alanine, (1-amino-cyclopropyl)carboxylic acid, ( 1-aminocyclobutyl)carboxylic acid, (1-aminocyclopentyl)carboxylic acid, (1-aminocyclohexyl)carboxyl, (1-aminocycloheptyl)carboxylic acid or (1-aminocyclooctyl)carboxylate Acid; has been reported for the amino acid sequence of GLP (Lopez, LC et al., Proc. Nat'l, Acad. Sci., USA 80 , 5485-5489, 1983; Bell, GI, et al., Nature 302 : 716- 718 (1983); Heinrich, G., et al, Endocrinol, 115 : 2176-2181 (1984). The structure of the proglucagonogen mRNA and its corresponding amino acid sequence are known to the public. The precursor gene product and proglucagon hydrolyzed into glucagon and insulinotropic peptides were characterized. The GLP-1 (1-37) label described herein refers to all from 1 position (N -end) a GLP-1 polypeptide to amino acid at position 37 (C-terminal). Similarly, the labeling of GLP-1 (7-37) described herein refers to all from the 7 position (N- a GLP-1 polypeptide to amino acid at position 37 (C-terminal). Similarly, the labeling of GLP-1 (7-36) as used herein refers to all from 7 (N-terminal) to 36 (C) - a terminal amino acid GLP-1 polypeptide.

本發明的另一個目的是提供一種藥物組合物，含有至少一種本發明的化合物和一種或一種以上藥學上可接受的載體、稀釋劑或輔料。Another object of the invention is to provide a pharmaceutical composition comprising at least one compound of the invention and one or more pharmaceutically acceptable carriers, diluents or adjuvants.

多肽的固相合成原理是本領域公知的，也可在本領域的通用出版物中查閱到，例如：Dugas,H. and Penney,C.,Bioorganic Chemistry(1981) Springer-Verlag,New York,page 54-92;Merrifield,J.M.,Chem. Soc.,85 ,2149,1962,and Stewart and Young,Solid Phase Peptide Synthesis,page 24-66,Freeman(San Francisco,1969)。The principle of solid phase synthesis of polypeptides is well known in the art and can also be found in general publications in the art, for example: Dugas, H. and Penney, C., Bioorganic Chemistry (1981) Springer-Verlag, New York, page 54-92; Merrifield, JM, Chem. Soc., 85 , 2149, 1962, and Stewart and Young, Solid Phase Peptide Synthesis, page 24-66, Freeman (San Francisco, 1969).

例如，本發明提供的肽片段可採用430肽合成儀(Applied Biosystems,Inc.,850 Lincoln Centre Drive,Foster City,CA 94404(應用生物系統有限公司，林肯中心區850號，福斯特市，加利福利亞州，94404))和應用生物系統有限公司提供的合成裝置的固相合成法得到。受Boc保護的氨基酸和試劑可從應用生物系統有限公司或其他化學品供應商購得。使用雙聯方案的連續的Boc化學方法可應用於對-甲基二苯甲胺樹脂，從而產生C-末端的羧醯胺。為了得到C-末端的酸，可以使用相應的PAM樹脂。可使用經預處理的羥基苯並***酯使天冬氨酸、谷氨醯胺和精氨酸進行偶聯。For example, the peptide fragment provided by the present invention can be used as a 430 peptide synthesizer (Applied Biosystems, Inc., 850 Lincoln Centre Drive, Foster City, CA 94404 (Applied Biosystems Co., Ltd., Lincoln Center Area 850, Foster City, Canada) Lieutenant State, 94404)) and solid phase synthesis of synthetic devices supplied by Applied Biosystems. The Boc protected amino acids and reagents are commercially available from Applied Biosystems, Inc. or other chemical suppliers. A continuous Boc chemistry using a duplex scheme can be applied to the p-methylbenzhydrylamine resin to produce a C-terminal carboguanamine. In order to obtain the C-terminal acid, the corresponding PAM resin can be used. The aspartic acid, glutamine and arginine can be coupled using a pretreated hydroxybenzotriazole ester.

本發明的又一目的是提供一種含有至少一種本發明化合物的藥物製劑，其含有濃度0.1mg/ml~25mg/ml的至少一種本發明的化合物，其中所述藥物製劑的pH是3.0~9.0。所述製劑還進一步包括緩衝溶液系統、防腐劑、滲透劑、螯合劑、穩定劑和表面活性劑。在本發明的一個具體實施方案中，所述藥物製劑室一種水狀製劑，例如含水製劑。所述製劑優選溶液或懸浮液。在本發明的另一個實施方案中，所述藥物製劑是一種水溶液。本文中術語“水狀製劑”是指含有至少50%w/w水的製劑。類似地，本文中術語“水溶液”是指含有至少50%w/w水的溶液，術語“水懸浮劑”是指含有至少50%w/w水的懸浮液。A further object of the present invention is to provide a pharmaceutical preparation comprising at least one compound of the invention comprising at least one compound of the invention at a concentration of from 0.1 mg/ml to 25 mg/ml, wherein the pH of the pharmaceutical preparation is from 3.0 to 9.0. The formulation further includes a buffer solution system, a preservative, a penetrant, a chelating agent, a stabilizer, and a surfactant. In a particular embodiment of the invention, the pharmaceutical preparation chamber is an aqueous preparation, such as an aqueous preparation. The formulation is preferably a solution or suspension. In another embodiment of the invention, the pharmaceutical formulation is an aqueous solution. The term "aqueous formulation" as used herein refers to a formulation containing at least 50% w/w water. Similarly, the term "aqueous solution" as used herein refers to a solution containing at least 50% w/w water, and the term "aqueous suspension" refers to a suspension containing at least 50% w/w water.

在本發明的另一個實施方案中，所述藥物製劑是凍幹製劑，在使用前醫師或者患者需要加入溶劑和/或稀釋劑。In another embodiment of the invention, the pharmaceutical formulation is a lyophilized formulation, and the physician or patient needs to add a solvent and/or diluent prior to use.

在本發明的另一個實施方案中，所述藥物製劑是乾燥製劑如凍幹或噴霧乾燥製劑，在使用前無需溶解。In another embodiment of the invention, the pharmaceutical formulation is a dry formulation such as a lyophilized or spray dried formulation which does not require dissolution prior to use.

在本發明的另一個實施方案中，本發明涉及一種含有至少一種本發明化合物的水狀製劑和緩沖劑的藥物製劑，含有濃度0.1mg/ml或以上的至少一種本發明化合物，其pH為3.0~9.0。在本發明的另一個實施例中，所述製劑的pH優選為7.0~9.5。在本發明的另一個實施例中，所述製劑的pH為3.0~7.0。在本發明的另一個實施例中，所述製劑的pH為5.0~7.5。在本發明的另一個實施例中，所述製劑的pH為7.5~9.0。在本發明的另一個實施例中，所述製劑的pH為7.5~8.5。在本發明的另一個實施例中，所述製劑的pH為6.0~7.5。In another embodiment of the invention, the invention relates to a pharmaceutical preparation comprising at least one aqueous preparation of a compound of the invention and a buffer comprising at least one compound of the invention at a concentration of 0.1 mg/ml or more, having a pH of 3.0 ~9.0. In another embodiment of the invention, the pH of the formulation is preferably from 7.0 to 9.5. In another embodiment of the invention, the formulation has a pH of from 3.0 to 7.0. In another embodiment of the invention, the formulation has a pH of from 5.0 to 7.5. In another embodiment of the invention, the formulation has a pH of from 7.5 to 9.0. In another embodiment of the invention, the formulation has a pH of from 7.5 to 8.5. In another embodiment of the invention, the formulation has a pH of from 6.0 to 7.5.

在本發明的另一個實施例中，所述製劑的pH更優選為6.0~7.0。在另一個實施方案中，所述藥物製劑的pH為8.0~8.5。In another embodiment of the invention, the pH of the formulation is more preferably from 6.0 to 7.0. In another embodiment, the pharmaceutical formulation has a pH of from 8.0 to 8.5.

在本發明的一個優選實施方案中，所述緩衝劑選自醋酸鈉、碳酸鈉、檸檬酸鹽、雙甘肽、組氨酸、甘氨酸、賴氨酸、精氨酸、磷酸二氫鈉、磷酸氫二鈉、磷酸鈉和三(羥基甲基)-氨基甲烷、二甘氨酸、三甘氨酸、蘋果酸、琥珀酸鹽、馬來酸、富馬酸、酒石酸、天冬氨酸或它們的混合物。上述特定緩衝劑的每一個都可以組成本發明的選擇性的實施方案。In a preferred embodiment of the invention, the buffer is selected from the group consisting of sodium acetate, sodium carbonate, citrate, diglycine, histidine, glycine, lysine, arginine, sodium dihydrogen phosphate, phosphoric acid Disodium hydrogen, sodium phosphate and tris(hydroxymethyl)-aminomethane, diglycine, triglycine, malic acid, succinate, maleic acid, fumaric acid, tartaric acid, aspartic acid or a mixture thereof. Each of the above specific buffers may constitute an optional embodiment of the invention.

在本發明進一步優選實施方案中，所述製劑還進一步含有藥學上可接受防腐劑。在本發明的一個優選實施方案中，所述防腐劑選自苯酚、鄰-甲酚、間-甲酚、對-甲酚、對-羥基苯甲酸甲酯、對-羥基苯甲酸丙酯、2-苯氧基乙醇、對-羥基苯甲酸丁酯、2-苯基乙醇、苄基乙醇、氯丁醇、硫柳汞、溴硝醇、苯甲酸、咪唑烷脲、氯己啶、脫氫醋酸鈉、氯甲酚、對-羥基苯甲酸乙酯、鹽酸苯索氯銨、氯苯甘醚(3對-氯苯氧基丙烷-1,2-二醇)。在一個實施方案中，所述防腐劑選自苯酚或間甲酚。在本發明的一個優選實施方案中，所述防腐劑的濃度為0.1mg/ml~20mg/ml。在本發明的一個優選實施方案中，所述防腐劑的濃度為0.1mg/ml~5mg/ml。在本發明的一個優選實施方案中，所述防腐劑的濃度為5mg/ml~10mg/ml。在本發明的一個優選實施方案中，所述防腐劑的濃度為10mg/ml~20mg/ml。上述特定防腐劑的每一個都可以組成本發明的選擇性的具體實施方案。本發明藥物藥物組合物中使用的防腐劑是本領域專業技術人員熟知的。為了方便，可參考Remington: The Science and Practice of Pharmacy,19^th edition,1995。In a further preferred embodiment of the invention, the formulation further comprises a pharmaceutically acceptable preservative. In a preferred embodiment of the invention, the preservative is selected from the group consisting of phenol, o-cresol, m-cresol, p-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2 -phenoxyethanol, butyl p-hydroxybenzoate, 2-phenylethanol, benzyl alcohol, chlorobutanol, thimerosal, bronopol, benzoic acid, imidazolidine, chlorhexidine, sodium dehydroacetate, Chlorocresol, ethyl p-hydroxybenzoate, benzethonium chloride hydrochloride, chlorophenylglycol (3 p-chlorophenoxypropane-1,2-diol). In one embodiment, the preservative is selected from the group consisting of phenol or m-cresol. In a preferred embodiment of the invention, the preservative is present in a concentration from 0.1 mg/ml to 20 mg/ml. In a preferred embodiment of the invention, the preservative is present in a concentration from 0.1 mg/ml to 5 mg/ml. In a preferred embodiment of the invention, the preservative is present in a concentration from 5 mg/ml to 10 mg/ml. In a preferred embodiment of the invention, the preservative is present in a concentration from 10 mg/ml to 20 mg/ml. Each of the specific preservatives described above may constitute an optional embodiment of the invention. Preservatives for use in the pharmaceutical compositions of the invention are well known to those skilled in the art. For convenience, reference is Remington: The Science and Practice of Pharmacy , 19 th edition, 1995.

在本發明的另一個優選實施方案中，所述製劑還進一步含有至少一種滲透劑。在本發明的一個優選實施方案中，所述滲透劑選自鹽(如氯化鈉)、糖或糖醇、氨基酸(如L-甘氨酸、L-組氨酸、精氨酸、賴氨酸、異亮氨酸、天冬氨酸、色氨酸、蘇氨酸)、醛醇(如丙三醇(甘油)、1,2-丙二醇(丙烯基乙二醇)、1,3-丙二醇,1,3-丁二醇)、聚乙二醇(如PEG400)或它們的混合物。在一個實施方案中，所述滲透劑是丙二醇，任何糖，例如單糖、二糖或多糖，或水溶性葡聚糖，包括如果糖、葡萄糖、甘露糖、山梨糖、木糖、麥芽糖、乳糖、蔗糖、海藻糖、右旋糖酐、支鏈澱粉、糊精、環糊***溶性澱粉、羥乙基澱粉和羰基甲基-纖維素鈉。在一個實施方案中，所述糖添加劑是蔗糖。糖醇是指含有至少一個羥基的C₄ -C₈ 碳氫化合物，包括，例如甘露醇、山梨醇、肌醇、衛矛醇(galactitol)、己六醇(dulcitol)、木糖醇、***醇。在一個實施方案中，所述糖醇是甘露醇。上述糖或糖醇可單獨使用或聯合使用。只要該糖或糖醇在液體製劑中是可以溶解的，對完成本發明使用的方法的穩定性效果不會造成不利影響的前提下，不對糖或糖醇的用量進行固定限制。在一個實施方案中，所述糖或糖醇的濃度是大約1mg/ml到大約150mg/ml。在本發明的一個優選實施方案中，所述滲透劑的濃度是1mg/ml~50mg/ml。在本發明的一個優選實施方案中，所述滲透劑的濃度為1mg/ml~7mg/ml。在本發明的一個優選實施方案中，所述滲透劑的濃度為5mg/ml~7mg/ml。在本發明的一個優選實施方案中，所述滲透劑的濃度為8mg/ml~24mg/ml。在本發明的一個優選實施方案中，所述滲透劑的濃度為25mg/ml~50mg/ml。上述特定滲透劑的每一個都可以組成本發明的選擇性的具體實施方案。本發明藥物藥物組合物中使用的滲透劑是本領域專業技術人員熟知的。為了方便，可參考Remington: The Science and Practice of Pharmacy,19^th edition,1995。In another preferred embodiment of the invention, the formulation further comprises at least one penetrant. In a preferred embodiment of the invention, the penetrant is selected from the group consisting of a salt (such as sodium chloride), a sugar or a sugar alcohol, an amino acid (such as L-glycine, L-histidine, arginine, lysine, Isoleucine, aspartic acid, tryptophan, threonine), aldol (such as glycerol (glycerol), 1,2-propanediol (propylene glycol), 1,3-propanediol, 1 , 3-butanediol), polyethylene glycol (such as PEG400) or a mixture thereof. In one embodiment, the penetrant is propylene glycol, any sugar, such as a monosaccharide, disaccharide or polysaccharide, or a water soluble glucan, including if sugar, glucose, mannose, sorbose, xylose, maltose, lactose , sucrose, trehalose, dextran, amylopectin, dextrin, cyclodextrin water-soluble starch, hydroxyethyl starch and carbonylmethyl-cellulose sodium. In one embodiment, the sugar additive is sucrose. The sugar alcohol refers to a C ₄ -C ₈ hydrocarbon containing at least one hydroxyl group, including, for example, mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, arabitol . In one embodiment, the sugar alcohol is mannitol. The above sugars or sugar alcohols may be used singly or in combination. As long as the sugar or sugar alcohol is soluble in the liquid preparation, the amount of the sugar or sugar alcohol is not fixedly limited without adversely affecting the stability effect of the method used in the present invention. In one embodiment, the concentration of the sugar or sugar alcohol is from about 1 mg/ml to about 150 mg/ml. In a preferred embodiment of the invention, the concentration of the penetrant is from 1 mg/ml to 50 mg/ml. In a preferred embodiment of the invention, the penetrant is present in a concentration from 1 mg/ml to 7 mg/ml. In a preferred embodiment of the invention, the concentration of the penetrant is from 5 mg/ml to 7 mg/ml. In a preferred embodiment of the invention, the concentration of the penetrant is from 8 mg/ml to 24 mg/ml. In a preferred embodiment of the invention, the penetrant is present in a concentration from 25 mg/ml to 50 mg/ml. Each of the above specific penetrants may constitute an optional embodiment of the invention. Penetrants for use in the pharmaceutical compositions of the invention are well known to those skilled in the art. For convenience, reference is Remington: The Science and Practice of Pharmacy , 19 th edition, 1995.

在本發明的另一個優選實施方案中，所述製劑還進一步含有一種螯合劑。在本發明的一個優選實施方案中，所述螯合劑選自乙二胺四乙酸(EDTA)的鹽、檸檬酸、天冬氨酸或它們的混合物。在另一個優選實施方案中，所述螯合劑的濃度為0.1mg/ml~5mg/ml。在本發明的一個優選實施方案中，所述螯合劑的濃度為0.1mg/ml~2mg/ml。在本發明的一個優選實施方案中，所述螯合劑的濃度為2mg/ml~5mg/ml。上述特定螯合劑的每一個都可以組成本發明的選擇性的具體實施方案。本發明藥物藥物組合物中使用的螯合劑是本領域專業技術人員熟知的。為了方便，可參考Remington: The Science and Practice of Pharmacy,19^th edition,1995。In another preferred embodiment of the invention, the formulation further comprises a chelating agent. In a preferred embodiment of the invention, the chelating agent is selected from the group consisting of salts of ethylenediaminetetraacetic acid (EDTA), citric acid, aspartic acid or mixtures thereof. In another preferred embodiment, the concentration of the chelating agent is from 0.1 mg/ml to 5 mg/ml. In a preferred embodiment of the invention, the concentration of the chelating agent is from 0.1 mg/ml to 2 mg/ml. In a preferred embodiment of the invention, the concentration of the chelating agent is from 2 mg/ml to 5 mg/ml. Each of the above specific chelating agents may constitute an optional embodiment of the invention. Chelating agents for use in the pharmaceutical compositions of the invention are well known to those skilled in the art. For convenience, reference is Remington: The Science and Practice of Pharmacy , 19 th edition, 1995.

在本發明的另一個優選實施方案中，所述製劑還進一步含有一種穩定劑。本發明藥物組合物中使用的穩定劑是本領域專業技術人員熟知的。為了方便，可參考Remington: The Science and Practice of Pharmacy,19^th edition,1995。In another preferred embodiment of the invention, the formulation further comprises a stabilizer. Stabilizers for use in the pharmaceutical compositions of the invention are well known to those skilled in the art. For convenience, reference is Remington: The Science and Practice of Pharmacy , 19 th edition, 1995.

更具體的，本發明的組合物是穩定的液體藥物組合物。所述液體藥物組合物的治療活性成分包括能夠在液體藥物製劑儲存中表現出聚合體形態的多肽。下文中的術語“形成聚集物”是指導致形成低聚物的多肽分子間是物理作用，這種低聚物仍然可溶，或肉眼可見聚集物從溶液中沉澱出來。下文中的術語“在存儲中”是指液體藥物組合物或一次性製備的製劑，不立即施用於對象。相當地，下述製劑可以液體形式、冷凍形態或乾燥形式包裝用以存儲，從而在後期處理成液體形式或其他適用於物件給藥的形式。術語“乾燥形態”是指液體藥物組合物或製劑採用凍幹法(例如凍幹法，參見Williams and PoIIi(1984) J. Parenteral Sci. Technol. 38:48-59)、噴霧乾燥法(參見Masters(1991) in Spray-Drying Handbook(5th ed;Longman Scientific and Technical,Essez,U.K.),pp. 491-676;Broadhead et al.(1992) Drug Devel. Ind. Pharm. 18:1 169-1206;and Mumenthaler et al.(1994) Pharm. Res. 11 :12-20)或空氣幹化法(Carpenter and Crowe(1988) Cryobiology 25:459-470;and Roser(1991) Biopharm. 4:47-53)進行乾燥。液體製劑藥物組合物的多肽在存儲中形成的聚集物能夠對該多肽的生物活性造成不利影響，導致該藥物組合物的治療效果損失。進一步，聚集物形成還可能引起其他問題，例如在採用輸注系統施用含有該多肽的藥物組合物時，該聚集物形態會堵塞針管、濾膜或輸液泵。More specifically, the compositions of the present invention are stable liquid pharmaceutical compositions. The therapeutically active ingredient of the liquid pharmaceutical composition comprises a polypeptide which is capable of exhibiting a polymeric form in the storage of a liquid pharmaceutical preparation. The term "forming aggregates" hereinafter refers to the physical interaction between the molecules of the polypeptide that result in the formation of the oligomer, the oligomers are still soluble, or the macroscopic aggregates are precipitated from the solution. The term "in storage" hereinafter refers to a liquid pharmaceutical composition or a one-time preparation that is not immediately administered to a subject. Quite, the formulations described below may be packaged for storage in liquid form, in frozen form or in dry form for later processing into a liquid form or other form suitable for administration of the article. The term "dry form" means that the liquid pharmaceutical composition or formulation is subjected to lyophilization (for example, lyophilization, see Williams and PoIIi (1984) J. Parenteral Sci. Technol. 38: 48-59), spray drying (see Masters). (1991) in Spray-Drying Handbook (5th ed; Longman Scientific and Technical, Essez, UK), pp. 491-676; Broadhead et al. (1992) Drug Devel. Ind. Pharm. 18:1 169-1206; Mumenthaler et al. (1994) Pharm. Res. 11: 12-20) or air drying (Carpenter and Crowe (1988) Cryobiology 25: 459-470; and Roser (1991) Biopharm. 4: 47-53) dry. The aggregate formed by the polypeptide of the liquid formulation pharmaceutical composition in storage can adversely affect the biological activity of the polypeptide, resulting in a loss of therapeutic effect of the pharmaceutical composition. Further, aggregate formation may also cause other problems, such as when the pharmaceutical composition containing the polypeptide is administered using an infusion system, the aggregate morphology can clog the needle, filter or infusion pump.

本發明的藥物組合物還可以進一步含有一定劑量的氨基酸堿，該氨基酸堿通過該組合物的多肽存儲過程，足以降低聚集物形成。術語“氨基酸堿”是指一種氨基酸或氨基酸聯用，其中任意使用的氨基酸既可以是其游離堿形式也可以是其鹽形式。當使用氨基酸聯用時，所有的氨基酸可以是其游離堿或鹽形式，或者當其他氨基酸為鹽形式時，也可以部分使用氨基酸游離堿。在本發明的一個實施方案中，用於製備本發明組合物的氨基酸是帶電荷側鏈的，例如，精氨酸、賴氨酸、天冬氨酸、谷氨酸。本發明的藥物組合物可使用某些特定的氨基酸(如蛋氨酸、組氨酸、咪唑、精氨酸、異亮氨酸、天冬氨酸、色氨酸、蘇氨酸或它們的混合物)的任意立體異構體(如L構型、D構型或它們的混合物)或這些異構體的混合物，其前提條件是只要該特定氨基酸是其游離堿或其鹽形式。在本發明的一個實施方案中，所述氨基酸是L-立體異構體。本發明的組合物也可以使用這些氨基酸類似物進行製備。術語“氨基酸類似物”是指天然存在的氨基酸的衍生物，這些氨基酸能夠達到降低本發明液體藥物組合物在存儲中的多肽形成聚集物的理想效果。適宜的精氨酸類似物包括，例如氨基胍、鳥氨酸和N-單乙基L-精氨酸，適宜的蛋氨酸類似物包括乙硫氨基酪酸和丁硫氨酸，適宜的半胱氨酸類似物包括S-甲基-L半胱氨酸。與其他氨基酸類似，本發明的組合物也含有上述氨基酸類似物的游離堿或鹽形式。在本發明的一個優選實施方案中，上述氨基酸或氨基酸衍生物的濃度能夠充分地阻止或延緩蛋白的聚集過程。在本發明的一個優選實施方案中，當多肽治療活性成分是含有至少一個易氧化的蛋氨酸片段時，可添加蛋氨酸(或其他含硫的氨基酸或氨基酸類似物)以抑制蛋氨酸片段氧化成蛋氨酸亞碸。術語“抑制”是指將整個過程中蛋氨酸氧化物的累積量最小化。抑制蛋氨酸氧化反應能夠將多肽最大限度地保持在適當的分子形態。可使用任意的蛋氨酸立體異構體(L或D構型)或它們的混合物。可添加一定量的蛋氨酸立體異構體或混合物以抑制蛋氨酸殘基的氧化作用，從而使得蛋氨酸亞碸的含量對調節試劑是可以接受的。典型地，這意味著本發明的藥物組合物含有不超過約10%到約30%的蛋氨酸亞碸。通常，可以通過加入蛋氨酸來完成上述要求，至此加入的蛋氨酸與蛋氨酸殘基的用量範圍比約是1:1到約1000:1，例如約10:1到約100:1。The pharmaceutical compositions of the present invention may further comprise a dose of an amino acid oxime sufficient to reduce aggregate formation by the polypeptide storage process of the composition. The term "amino acid oxime" refers to an amino acid or a combination of amino acids, wherein any amino acid used may be either in its free oxime form or in its salt form. When a combination of amino acids is used, all of the amino acids may be in the form of their free oxime or salt, or when other amino acids are in the form of a salt, the amino acid free oxime may also be used in part. In one embodiment of the invention, the amino acid used to prepare the compositions of the invention is a charged side chain, for example, arginine, lysine, aspartic acid, glutamic acid. The pharmaceutical composition of the present invention may use certain specific amino acids (such as methionine, histidine, imidazole, arginine, isoleucine, aspartic acid, tryptophan, threonine or a mixture thereof). Any stereoisomer (such as the L configuration, the D configuration or a mixture thereof) or a mixture of these isomers, provided that the particular amino acid is in the form of its free oxime or its salt. In one embodiment of the invention, the amino acid is an L-stereoisomer. Compositions of the invention may also be prepared using these amino acid analogs. The term "amino acid analog" refers to derivatives of naturally occurring amino acids which are capable of achieving the desired effect of reducing the formation of aggregates of polypeptides in storage by the liquid pharmaceutical compositions of the present invention. Suitable arginine analogs include, for example, aminoguanidine, ornithine and N-monoethyl L-arginine, suitable methionine analogs include ethylthiobutyric acid and butyl methionine, suitable cysteines Analogs include S-methyl-L cysteine. Similar to other amino acids, the compositions of the present invention also contain the free oxime or salt form of the above amino acid analogs. In a preferred embodiment of the invention, the concentration of the above amino acid or amino acid derivative is sufficient to prevent or delay the aggregation process of the protein. In a preferred embodiment of the invention, when the polypeptide therapeutically active ingredient is a fragment comprising at least one oxidizable methionine, methionine (or other sulfur-containing amino acid or amino acid analog) may be added to inhibit oxidation of the methionine fragment to methionine. . The term "inhibiting" means minimizing the cumulative amount of methionine oxide throughout the process. Inhibition of methionine oxidation allows the polypeptide to be held in the proper molecular form to the maximum extent possible. Any methionine stereoisomer (L or D configuration) or a mixture thereof can be used. A certain amount of a methionine stereoisomer or mixture may be added to inhibit the oxidation of the methionine residue, such that the content of methionine hydrazide is acceptable for the conditioning reagent. Typically, this means that the pharmaceutical compositions of the present invention contain no more than about 10% to about 30% methionine hydrazide. In general, the above requirements can be accomplished by the addition of methionine, whereby the amount of methionine to methionine residue added ranges from about 1:1 to about 1000:1, such as from about 10:1 to about 100:1.

在本發明的一個優選實施方案中，所述製劑還進一步含有穩定劑，選自高分子量的聚合物或低分子量的分子化合物。在本發明的一個優選實施方案中，所述穩定劑選自聚乙二醇(如PEG 3350)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮、羰基纖維素或羥基纖維素或它們的衍生物(如HPC、HPC-SL、HPC-L、HPMC)、環糊精、含硫物質(如硫代甘油、硫代乙醇酸、2-甲基硫代乙醇)或不同的鹽(如氯化鈉)。上述特定穩定劑的每一個都可以組成本發明的選擇性的具體實施方案。In a preferred embodiment of the invention, the formulation further comprises a stabilizer selected from the group consisting of high molecular weight polymers or low molecular weight molecular compounds. In a preferred embodiment of the invention, the stabilizer is selected from the group consisting of polyethylene glycol (such as PEG 3350), polyvinyl alcohol (PVA), polyvinylpyrrolidone, carbonyl cellulose or hydroxy cellulose or derivatives thereof ( Such as HPC, HPC-SL, HPC-L, HPMC), cyclodextrin, sulfur-containing substances (such as thioglycerol, thioglycolic acid, 2-methylthioethanol) or different salts (such as sodium chloride) . Each of the above specific stabilizers may constitute an optional embodiment of the invention.

所述藥物組合物也可以含有其他穩定劑，進一步增強治療活性成分多肽的穩定性。具體對本發明有意義的穩定劑，包括但不限於：蛋氨酸和EDTA，用於阻止多肽蛋氨酸氧化反應；一種非離子表面活性劑，用於阻止多肽凍融或機械剪切過程中伴隨的聚集作用。The pharmaceutical compositions may also contain other stabilizers to further enhance the stability of the therapeutically active ingredient polypeptide. Stabilizers that are particularly useful for the present invention include, but are not limited to, methionine and EDTA for blocking the oxidation of the polypeptide methionine; a nonionic surfactant for preventing the concomitant aggregation associated with freeze-thaw or mechanical shearing of the polypeptide.

在本發明的另一個優選實施方案中，所述製劑還進一步含有表面活性劑。在本發明的另一個具體實施方案中，所述藥物組合物含有兩種不同的表面活性劑。本文中術語“表面活性劑”是指任意含有水溶性(親水性)部分、頭部基團和脂溶性(親脂性)部分的分子或離子。表面活性劑優選在介面聚集，其親水性部分指向水相(親水相)、親脂性部分指向油相疏水相(如玻璃、空氣、油等)。表面活性劑開始形成離子膠束的濃度定義為臨界膠束濃度(critical micelle concentration,CMC)。此外，表面活性劑的表面張力低於液體表面張力。表面活性劑是熟知的兩性化合物。通常情況下，術語“清潔劑”可作為表面活性劑的同義詞使用。In another preferred embodiment of the invention, the formulation further comprises a surfactant. In another embodiment of the invention, the pharmaceutical composition contains two different surfactants. The term "surfactant" as used herein refers to any molecule or ion that contains a water soluble (hydrophilic) moiety, a head group, and a fat soluble (lipophilic) moiety. The surfactant preferably aggregates at the interface, the hydrophilic portion of which is directed to the aqueous phase (hydrophilic phase) and the lipophilic portion to the aqueous phase of the oil phase (eg, glass, air, oil, etc.). The concentration at which the surfactant begins to form ionic micelles is defined as the critical micelle concentration (CMC). In addition, the surface tension of the surfactant is lower than the surface tension of the liquid. Surfactants are well known amphoteric compounds. In general, the term "cleanser" can be used synonymously with surfactants.

陰離子表面活性劑選自：鵝脫氧膽酸、鵝脫氧膽酸鈉、膽酸、脫氫膽酸、脫氧膽酸、脫氧膽酸甲酯、洋地黃皂苷、洋地黃毒苷元、N,N-二甲基十二胺-N-氧化物、多庫酯鈉、甘氨鵝去氧膽酸鈉、甘氨膽酸水合物、甘氨脫氧膽酸一水合物、甘氨脫氧膽酸鈉、甘氨石膽酸-3-硫酸二鈉鹽、甘氨石膽酸乙酯、N-十二烷基肌氨酸鈉、N-十二烷基肌氨酸、十二烷基硫酸鋰、盧戈耳溶液(Lugol)、1-辛基磺酸鈉、1-丁基磺酸鈉、1-癸烷磺酸鈉、1-十二烷磺酸鈉、1-庚烷磺酸鈉、1-壬烷磺酸鈉、1-丙烷磺鈉一水合物、2-溴乙烷磺酸鈉、膽酸鈉水合物、牛膽汁或羊膽汁、膽汁酸鈉水合物、絡膽酸鈉、脫氧膽酸鈉、十二烷基硫酸鈉、己烷磺酸鈉、辛基磺酸鈉、丁烷磺酸鈉、牛黃膽酸鈉、牛磺鵝去氧膽酸鈉、牛磺鵝去氧膽酸鈉一水合物、牛磺鵝去氧膽酸-3-硫酸二鈉鹽、牛磺熊去氧膽酸鈉、十二烷基磺酸、多庫酯鈉(docusate sodium,DSS,CAS註冊號[577-11-7])、多庫酯鈣(CAS註冊號[128-49-4]),多庫酯鉀(CAS註冊號[7491-09-0])、十二烷基磺酸鈉(sodium dodecyl sulfate,sodium lauryl sulfate,SDS)、十二烷基磷酸膽鹼(Dodecylphosphocholine,FOS-Choline-12)、癸基磷酸膽鹼(Decylphosphocholine,FOS-Choline-10)、壬基磷酸膽鹼(Nonylphosphocholine,FOS-Choline-9)、二棕櫚醯磷脂酸、辛酸鈉和/或熊去氧膽酸。The anionic surfactant is selected from the group consisting of: chenodeoxycholic acid, sodium chenodeoxycholate, cholic acid, dehydrocholic acid, deoxycholic acid, methyl deoxycholate, digitonin, digitoxin, N, N - dimethyldodecyl-N-oxide, sodium docusate, sodium sulphate deoxycholate, glycocholic acid hydrate, glycodeoxycholic acid monohydrate, sodium glycodeoxycholate, Glycine cholic acid-3-sulfate disodium salt, glycine bile acid ethyl ester, N-dodecyl sarcosinate sodium, N-dodecyl sarcosine, lithium lauryl sulfate, Lu Lugol, sodium 1-octylsulfonate, sodium 1-butylsulfonate, sodium 1-decanesulfonate, sodium 1-dodecanesulfonate, sodium 1-heptanesulfonate, 1- Sodium decane sulfonate, sodium 1-propane sulfonate monohydrate, sodium 2-bromoethane sulfonate, sodium cholate hydrate, bovine bile or sheep bile, sodium bile hydrate, sodium cholesteric acid, deoxycholic acid Sodium, sodium lauryl sulfate, sodium hexanesulfonate, sodium octyl sulfonate, sodium butane sulfonate, sodium taurocholate, sodium tauroche-deoxycholate, sodium tauro-sodeoxycholate Hydrate, taurochenodeoxycholic acid-3-sulfate disodium salt, taurours sodium deoxycholate , Dodecyl sulfonic acid, docusate sodium (DSS, CAS registration number [577-11-7]), docusate calcium (CAS registration number [128-49-4]), potassium docusate (CAS registration number [7491-09-0]), sodium dodecyl sulfate (sodium lauryl sulfate, SDS), dodecylphosphocholine (FOS-Choline-12), 癸Decylphosphocholine (FOS-Choline-10), Nonylphosphocholine (FOS-Choline-9), dipalmitoside phosphatidic acid, sodium octanoate and/or ursodeoxycholic acid.

陽離子表面活性劑選自：烷基三甲基溴化銨、苯甲烷氯化銨、苄基二甲基十六烷基氯化銨、苄基二甲基十四烷基氯化銨、苄基二甲基十四烷基四氯化碘銨、二甲基雙十八烷基溴化銨、十二烷基乙基二甲基溴化銨、十二烷基三甲基溴化銨、乙基十六烷基二甲基溴化銨、十六烷基三甲基溴化銨、聚氧化乙烯(10)-N-牛油-1,3-二氨基丙烷、十六基/溴化二銨和/或三甲基十四烷基溴化銨。The cationic surfactant is selected from the group consisting of alkyltrimethylammonium bromide, benzylmethane chloride, benzyldimethylhexadecyl ammonium chloride, benzyldimethyltetradecyl ammonium chloride, benzyl Dimethyltetradecyl ammonium iodide, dimethyl dioctadecyl ammonium bromide, dodecyl ethyl dimethyl ammonium bromide, dodecyl trimethyl ammonium bromide, B Cetyldimethylammonium bromide, cetyltrimethylammonium bromide, polyoxyethylene (10)-N-tallow-1,3-diaminopropane, hexadecyl/brominated Ammonium and/or trimethyltetradecyl ammonium bromide.

非離子表面活性劑選自：BigCHAP，雙(聚乙烯乙二醇二咪坐基羰基)，嵌段共聚物例如聚氧化乙烯，或聚氧化丙烯嵌段共聚物如泊洛沙姆、泊洛沙姆-188和泊洛沙姆-407、35、56,72,76、92V、97、58P、EL、月桂醇聚氧乙烯醚、N-癸醯基-N-甲基-葡糖胺、n-十二醯基-N-甲基葡糖胺、烷基-聚葡萄糖苷、乙氧基化的蓖麻油、七乙二醇單癸醚、七乙二醇單十二醚、七乙二醇單十四醚、六乙二醇單十二醚、六乙二醇單十六醚、六乙二醇十八醚、六乙二醇十四醚、lgepal CA-630、lgepal CA-630、甲基-6-0-(N-庚基氨基甲醯)-β-D-葡萄吡喃糖甙、就乙二醇單十二醚、N-壬醯基-N-甲基葡糖胺、N-壬醯基-N-甲基葡糖胺、八乙二醇單癸醚、八乙二醇單十二醚、八乙二醇單十六醚、八乙二醇單十八醚、八乙二醇單十四醚、八-β-D-葡萄吡喃糖甙、五乙二醇單癸醚、五乙二醇單十二醚、五乙二醇單十六醚、五乙二醇單己醚、五乙二醇單十八醚、五乙二醇單辛醚、聚乙二醇二環氧丙醚、聚乙二醇醚W-1、聚氧化乙烯10十三醚、聚氧化乙烯100硬脂酸酯、聚氧化乙烯20異十六醚、聚氧化乙烯20油烯醚、聚氰乙烯40硬脂酸酯、聚氧化乙烯50硬脂酸酯、聚氧化乙烯8硬脂酸酯、聚氧化乙烯雙(咪唑基羰基)、聚氧化乙烯25丙二醇硬脂酸酯、皂皮樹(Quillaja bark)提取物皂素、司20(20)、司40、司60、司65、司80、司85、Tergitol Type 15-S-12、Tergitol Type 15-S-30、Tergitol Type 15-S-5、Tergitol Type 15-S-7、Tergitol Type 15-S-9、Tergitol Type NP-10、Tergitol Type NP-4、Tergitol Type NP-40、Tergitol Type NP-7、Tergitol Type NP-9、十四烷基-β-D-麥芽糖苷、四乙二醇單癸醚、四乙二醇單十二醚、四乙二醇單十四醚、三乙二醇單癸醚、三乙二醇單十二醚、三乙二醇單十六醚、三乙二醇單辛醚、三乙二醇單十四醚、Triton CF-21、Triton CF-32、Triton DF-12、Triton DF-16、Triton GR-5M、Triton QS-15、Triton QS-44、Triton X-100、Triton X-102、Triton X-15、Triton X-151、Triton X-200、Triton X-207、X-100、X-114、X-165溶液、X-305溶液、X-405、X-45、X-705-70、吐20(20)、吐40、吐60、吐6、吐65、吐80、吐81、吐85、泰羅沙伯、鞘磷脂(sphingophospholipids,sphingomyelin)、鞘糖脂(sphingoglycolipids,神經醯胺ceramides、神經節苷脂gangliosides)、磷脂、和/或n-十一烷基-β-D-葡萄吡喃糖甙。The nonionic surfactant is selected from the group consisting of: BigCHAP, bis(polyethylene glycol dimercaptocarbonyl), block copolymers such as polyethylene oxide, or polyoxypropylene block copolymers such as poloxamer, poloxa M-188 and Polosham-407, 35. 56, 72, 76. 92V, 97. 58P, EL, lauryl polyoxyethylene ether, N-mercapto-N-methyl-glucosamine, n-dodedecyl-N-methylglucamine, alkyl-polyglucoside, ethoxylation Castor oil, heptaethylene glycol monoterpene ether, heptaethylene glycol monododecyl ether, heptaethylene glycol monotetradecyl ether, hexaethylene glycol monododecyl ether, hexaethylene glycol monohexadecyl ether, six ethyl Glycol octadecyl ether, hexaethylene glycol tetradecyl ether, lgepal CA-630, lgepal CA-630, methyl-6-0-(N-heptylaminocarboxamidine)-β-D-glucopyranose Ethylene glycol monododecyl ether, N-mercapto-N-methylglucamine, N-mercapto-N-methylglucamine, octaethylene glycol monoterpene ether, octaethylene glycol Monododecyl ether, octaethylene glycol monohexadecyl ether, octaethylene glycol monostearyl ether, octaethylene glycol monotetradecyl ether, octa-β-D-glucopyranose, pentaethylene glycol monoterpene Ether, pentaethylene glycol monododecyl ether, pentaethylene glycol monohexadecyl ether, pentaethylene glycol monohexyl ether, pentaethylene glycol monostearyl ether, pentaethylene glycol monooctyl ether, polyethylene glycol II Glycidyl ether, polyethylene glycol ether W-1, polyoxyethylene tridecyl ether, polyoxyethylene 100 stearate, polyoxyethylene 20 isohexadecyl ether, polyoxyethylene 20 olefinene , polycyanoethylene 40 stearate, polyoxyethylene 50 stearate, polyoxyethylene 8 stearate, polyethylene oxide bis(imidazolylcarbonyl), polyethylene oxide 25 propylene glycol stearate, soap hull Tree (Quillaja bark) extract saponin, division 20( 20), Division 40, Division 60, Division 65, Division 80, Division 85, Tergitol Type 15-S-12, Tergitol Type 15-S-30, Tergitol Type 15-S-5, Tergitol Type 15-S-7, Tergitol Type 15-S-9, Tergitol Type NP-10, Tergitol Type NP-4, Tergitol Type NP-40, Tergitol Type NP-7, Tergitol Type NP-9, tetradecyl-β-D-maltoside, tetraethylene glycol monoterpene ether, tetraethylene glycol monododecyl ether , tetraethylene glycol monotetradecyl ether, triethylene glycol monoterpene ether, triethylene glycol monododecyl ether, triethylene glycol monohexadecyl ether, triethylene glycol monooctyl ether, triethylene glycol mono Tetraether, Triton CF-21, Triton CF-32, Triton DF-12, Triton DF-16, Triton GR-5M, Triton QS-15, Triton QS-44, Triton X-100, Triton X-102, Triton X -15, Triton X-151, Triton X-200, Triton X-207, X-100, X-114, X-165 solution, X-305 solution, X-405, X-45, X-705-70, spit 20( 20), spit 40, spit 60, spit 6, spit 65, spit 80, spit 81, spit 85. Teresa, sphingomyelin, sphingomyelin, sphingoglycolipids, ceramides, gangliosides, phospholipids, and/or n-undecyl-β-D-glucopyridine Unsweetened.

兩性離子表面活性劑選自CHAPS，CHAPSO，3-(癸基二甲基氨基)丙磺酸內鹽，3-(十二烷基二甲基氨基)丙磺酸內鹽，3-(十二烷基二甲基氨基)丙磺酸內鹽，3-(N,N-二甲基肉豆蔻基氨基)丙磺酸鹽，3-(N,N-二甲基十八烷基氨基)-丙磺酸鹽，3-(N,N-二甲基辛基)丙磺酸內鹽，3-(N,N-二甲基棕櫚醯基氨基)丙磺酸鹽，N-烷基-N,N-二甲基氨基-1-丙磺酸鹽，3-膽醯胺-1-丙基二甲基氨基-1-丙磺酸鹽，十二烷基磷酸膽鹼，肉豆蔻醯基溶血卵磷脂醯膽鹼，Zwittergent 3-12(N-十二烷基-N,N-二甲基-3-氨基-1-丙磺酸鹽)，Zwittergent 3-10(3-(癸基二甲基氨基)-丙磺酸內鹽)，Zwittergent 3-08(3-(辛基二甲基氨基)丙磺酸鹽)，甘油磷脂(卵磷脂，腦磷脂，磷脂醯基絲氨酸)，甘油糖脂(吡喃半乳糖苷)，溶血磷脂醯膽鹼和磷脂醯膽鹼的烷基、烷氧基(烷基酯)、烷氧基(烷基醚)衍生物，例如溶血磷脂醯膽鹼的月桂醯和肉豆蔻醯衍生物、二棕櫚醯磷脂醯膽鹼和極性頭部基團的膽鹼類、乙醇胺類、磷脂酸、絲氨酸、蘇氨酸類、甘油、纖維醇、溶血磷脂醯絲氨酸和溶血性磷蘇氨酸、醯基肉鹼或其衍生物的修飾產物，賴氨酸、精氨酸或組氨酸的N^beta -醯基化衍生物，或賴氨酸或精氨酸的側鏈醯基化衍生物，含有賴氨酸、精氨酸或組氨酸和中性或酸性氨基酸任意組成的二肽的N^beta -醯基化衍生物，含有中性氨基酸和帶有兩個電荷的氨基酸任意組成的三肽的N^beta -醯基化衍生物；或者所述表面活性劑選自咪唑啉衍生物，C₆ -C₁₂ 長鏈脂肪酸或其鹽(如油酸和羊脂酸)，N-十六烷基-N,N-二甲基-3-氨基-1-丙磺酸鹽，陰離子的(烷基-芳香烴基-磺酸鹽)單價表面活性劑、棕櫚醯基溶血磷脂醯基-L-色氨酸、溶血磷脂類(如1-乙醇胺、膽鹼、絲氨酸、蘇氨酸的醯基-sn-甘油-3-磷酸酯)或它們的混合物。The zwitterionic surfactant is selected from the group consisting of CHAPS, CHAPSO, 3-(decyldimethylamino)propanesulfonic acid inner salt, 3-(dodecyldimethylamino)propanesulfonic acid inner salt, 3-(12 Alkyl dimethylamino)propane sulfonate, 3-(N,N-dimethylmyristylamino)propane sulfonate, 3-(N,N-dimethyloctadecylamino)- Propane sulfonate, 3-(N,N-dimethyloctyl)propane sulfonate, 3-(N,N-dimethylpalmitoylamino)propane sulfonate, N-alkyl-N , N-dimethylamino-1-propane sulfonate, 3-cholestyramine-1-propyldimethylamino-1-propane sulfonate, dodecylphosphocholine, myristyl hemolysis Lecithin choline, Zwittergent 3-12 (N-dodecyl-N,N-dimethyl-3-amino-1-propane sulfonate), Zwittergent 3-10 (3-(mercaptodimethyl dimethyl) Base amino)-propanesulfonic acid inner salt), Zwittergent 3-08 (3-(octyldimethylamino)propane sulfonate), glycerophospholipid (lecithin, cephalin, phospholipid thiol serine), glyceroglycolipid (galactopyranoside), lysophosphatidylcholine and phospholipid choline alkyl, alkoxy (alkyl ester), alkoxy (alkyl ether) derivatives, such as lysophosphatidylcholine Clams and nutmeg Derivatives, dipalmitoside phosphocholine choline and polar head group of choline, ethanolamine, phosphatidic acid, serine, threonine, glycerol, cellulose, lysophosphatidylcholine and hemolytic phosphothionine, a modified product of mercaptocarnitine or a derivative thereof, an N ^beta -thiolated derivative of lysine, arginine or histidine, or a side chain thiolated derivative of lysine or arginine, An N ^beta -thiolated derivative of a dipeptide comprising lysine, arginine or histidine and a neutral or acidic amino acid, containing a neutral amino acid and a tripeptide having an arbitrary composition of two charged amino acids N ^beta -thiolated derivative; or the surfactant is selected from the group consisting of imidazoline derivatives, C ₆ -C ₁₂ long chain fatty acids or salts thereof (such as oleic acid and lanolin), N-hexadecyl -N,N-dimethyl-3-amino-1-propane sulfonate, anionic (alkyl-aromatic hydrocarbon-sulfonate) monovalent surfactant, palmitoyl lysophosphatidyl thiol-L-color ammonia Acid, lysophospholipids (such as 1-ethanolamine, choline, serine, threonyl-sn-glycerol-3-phosphate of threonine) or mixtures thereof.

本文中術語“烷基-多苷”是指被一個或多個糖配基，例如麥芽糖苷、糖苷等取代的支鏈或支鏈的C_5-20 烷基、C_5-20 烯基或C_5-20 炔基鏈。上述烷基-多苷包括C_6-I8 alkyl-烷基-多苷。所述烷基-多苷的特定實施方案包括偶數個碳鏈，例如C₆ 烷基鏈、C₈ 烷基鏈、C₁₀ 烷基鏈、C₁₂ 烷基鏈、C₁₄ 烷基鏈、C₁₆ 烷基鏈、C₁₈ 烷基鏈和C₂₀ 烷基鏈。所述糖配基的具體實施方案包括吡喃糖苷、葡萄糖吡喃糖苷、麥芽糖苷、葡萄糖苷和蔗糖。在本發明的實施方案中，與烷基連接的糖配基數小於6個。在本發明的實施方案中，與烷基連接的糖配基數小於5個。在本發明的實施方案中，與烷基連接的糖配基數小於4個。在本發明的實施方案中，與烷基連接的糖配基數小於3個。在本發明的實施方案中，與烷基連接的糖配基數小於2個。烷基-多糖苷的具體實施方案是烷基葡萄糖苷類，例如n-癸基-β-D-葡萄吡喃糖苷、癸基-β-D-麥芽吡喃糖苷、十二烷基-β-D-葡萄吡喃糖苷、n-十二烷基-β-D-麥芽糖苷、n-十二烷基-β-D-麥芽糖苷、n-十二烷基-β-D-麥芽糖苷、十四烷基-β-D-葡萄吡喃糖苷、癸基-β-D-麥芽糖苷、十六烷基-β-D-麥芽糖苷、癸基-β-D-麥芽糖苷、十二烷基-β-D-麥芽三糖苷、十四烷基-β-D-麥芽三糖苷、十六烷基-β-D-麥芽三糖苷、n-十二烷基-蔗糖、n-癸基-蔗糖、蔗糖單矽酸酯、蔗糖單月桂酸酯、蔗單豆蔻酸酯和蔗糖棕櫚酸酯。本發明藥物藥物組合物中使用的表面活性劑是本領域專業技術人員熟知的。為了方便，可參考Remington: The Science and Practice of Pharmacy,19^th edition,1995。The term "alkyl-polyglycoside" as used herein refers to a branched or branched C _5-20 alkyl group, C _5-20 alkenyl group or C substituted by one or more sugar ligands such as maltosides, glycosides and the like. _5-20 alkynyl chain. The above alkyl-polyglycosides include C _6-I8 alkyl-alkyl-polyglycosides. Said alkyl - polyglycoside particular embodiment comprises an even number of carbon chains, e.g. C ₆ alkyl chain, C ₈ alkyl chains, C ₁₀ alkyl chains, C ₁₂ alkyl chains, C ₁₄ alkyl chains, C ₁₆ An alkyl chain, a C ₁₈ alkyl chain, and a C ₂₀ alkyl chain. Specific embodiments of the sugar ligands include pyranosides, glucopyranosides, maltosides, glucosides, and sucrose. In an embodiment of the invention, the number of sugar ligands attached to the alkyl group is less than 6. In an embodiment of the invention, the number of sugar ligands attached to the alkyl group is less than 5. In an embodiment of the invention, the number of glycosyl groups attached to the alkyl group is less than four. In an embodiment of the invention, the number of sugar ligands attached to the alkyl group is less than three. In an embodiment of the invention, the number of glycosyl groups attached to the alkyl group is less than two. A specific embodiment of the alkyl-polyglycoside is an alkyl glucoside such as n-fluorenyl-β-D-glucopyranoside, thiol-β-D-maltopyranoside, dodecyl-β -D-glucopyranoside, n-dodecyl-β-D-maltoside, n-dodecyl-β-D-maltoside, n-dodecyl-β-D-maltoside, Tetradecyl-β-D-glucopyranoside, thiol-β-D-maltoside, cetyl-β-D-maltoside, thiol-β-D-maltoside, dodecyl -β-D-maltotrioside, tetradecyl-β-D-maltotrioside, cetyl-β-D-maltotrioside, n-dodecyl-sucrose, n-癸Base-sucrose, sucrose monodecanoate, sucrose monolaurate, cane monostearate and sucrose palmitate. Surfactants for use in the pharmaceutical compositions of the invention are well known to those skilled in the art. For convenience, reference is Remington: The Science and Practice of Pharmacy , 19 th edition, 1995.

在本發明的一個優選實施方案中，所述製劑進一步含有蛋白酶抑制劑，例如乙二胺四乙酸(ethylenediamine tetraacetic acid,EDTA)和苄脒鹽酸鹽，以及其他可商業購得的蛋白酶抑制劑。在含有蛋白酶酵素原的藥物組合物中，使用蛋白酶抑制劑具有特定用途，可用於抑制自身催化反應。In a preferred embodiment of the invention, the formulation further comprises a protease inhibitor, such as ethylenediamine tetraacetic acid (EDTA) and benzamidine hydrochloride, as well as other commercially available protease inhibitors. In pharmaceutical compositions containing proproteinase, the use of protease inhibitors has particular utility and can be used to inhibit autocatalytic reactions.

本發明的肽藥物製劑還可以含有其他成分。上述添加組份可以包括潤濕劑、乳化劑、抗氧化劑、膨脹劑、滲透率改善劑、螯合劑、金屬離子、油性載體、蛋白(如人血清白蛋白、明膠和蛋白質)和兩性離子(例如氨基酸，如甜菜鹼、牛磺酸、精氨酸、甘氨酸、賴氨酸和組氨酸)。上述添加組份不應對本發明的藥物製劑的整體穩定性產生不利影響。The peptide pharmaceutical preparation of the present invention may further contain other components. The above additional components may include wetting agents, emulsifiers, antioxidants, bulking agents, permeability improvers, chelating agents, metal ions, oily carriers, proteins such as human serum albumin, gelatin and proteins, and zwitterions (eg Amino acids such as betaine, taurine, arginine, glycine, lysine and histidine). The above-mentioned added components should not adversely affect the overall stability of the pharmaceutical preparation of the present invention.

根據治療的需要，可從幾個部位對患者施用含有本發明化合物的藥物組合物，例如，典型部位如皮膚、粘膜，旁路吸收如動脈、血管、心臟，以及具有吸收作用的部位如皮膚、皮下、肌肉或腹腔。Depending on the needs of the treatment, the pharmaceutical composition containing the compound of the present invention can be administered to the patient from several sites, for example, typical sites such as skin, mucous membranes, bypass absorption such as arteries, blood vessels, heart, and areas having absorption such as skin, Subcutaneous, muscular or abdominal cavity.

根據病人的治療需要，本發明的藥物組合物的施用可通過幾種給藥途徑，例如舌頭、舌下、口腔、口腔內、口服、胃腸內、鼻、肺(如通過小支氣管、肺泡或它們的聯合)、表皮、真皮、經皮、***、直腸、眼睛(如通過結膜)、輸尿管和胃腸外等。Depending on the therapeutic needs of the patient, the pharmaceutical compositions of the invention may be administered by several routes of administration, such as the tongue, sublingual, buccal, buccal, oral, intragastric, nasal, pulmonary (eg, via the bronchioles, alveoli, or Combination), epidermis, dermis, percutaneous, vaginal, rectal, eye (eg through the conjunctiva), ureter and parenteral.

本發明的組合物可以按照幾種劑量形式施用，例如溶液、懸浮劑、乳劑、微乳、多重乳劑、泡沫劑、藥膏、糊劑、膏劑、軟膏劑、片劑、包衣片劑、洗劑、膠囊劑(如硬明膠膠囊和軟明膠膠囊)、栓劑、直腸用膠囊、滴劑、凝膠劑、噴霧劑、散劑、氣溶膠、吸入劑、滴眼液、眼膏、眼用清洗劑、子宮托、***環、***用藥膏、注射液、液液原位轉移(如原位凝膠的、原位安置、原位沉澱的、原位結晶的、輸液和植入體)。本發明的組合物還可以進一步，通過共價作用、疏水作用和靜電作用等方式混入或連接藥物載體，藥物分散系統和高級藥物分散系統，從而進一步達到增強穩定性、提高生物活性、增加溶解性、降低副作用、實現本領域技術人員熟知的時間療法、增加病人順應性的作用或上述作用的綜合。載體、藥物分散系統或高級藥物分散系統的具體實例選自，但不限於，高分子如纖維素或其衍生物，多糖如右旋糖酐或其衍生物，澱粉或其衍生物，聚乙烯醇，丙烯酸鹽聚合物和甲基丙烯酸酯聚合物，聚乳酸和聚乙醇酸和它們的嵌段聚合物，聚乙二烯醇，載體蛋白如血清，凝膠如熱凝膠化系統，如本領域技術人員熟知的共聚系統，膠束，脂質體，微球，納米粒子，液體晶體和它們的分散系，L2相和它們的分散系，本領域技術人員熟知油-水系統的相行為，高分子膠束，多重乳液，自乳化，在微乳化，環糊精或其衍生物，和樹形高分子。The composition of the present invention can be administered in several dosage forms, such as solutions, suspensions, emulsions, microemulsions, multiple emulsions, foams, ointments, pastes, ointments, ointments, tablets, coated tablets, lotions. Capsules (such as hard gelatin capsules and soft gelatin capsules), suppositories, rectal capsules, drops, gels, sprays, powders, aerosols, inhalants, eye drops, eye ointments, ophthalmic cleansers, Uterine support, vaginal ring, vaginal ointment, injection, in situ transfer of liquid (such as in situ gel, in situ placement, in situ precipitation, in situ crystallization, infusion and implant). The composition of the present invention may further further incorporate or link a drug carrier, a drug dispersion system and a high-grade drug dispersion system by covalent action, hydrophobic action and electrostatic action, thereby further enhancing stability, improving biological activity, and increasing solubility. The side effects are reduced, the time therapy well known to those skilled in the art is achieved, the effect of increasing patient compliance, or a combination of the above effects is achieved. Specific examples of the carrier, the drug dispersion system or the advanced drug dispersion system are selected from, but not limited to, a polymer such as cellulose or a derivative thereof, a polysaccharide such as dextran or a derivative thereof, starch or a derivative thereof, polyvinyl alcohol, acrylate Polymer and methacrylate polymers, polylactic acid and polyglycolic acid and their block polymers, polyvinylidene alcohol, carrier proteins such as serum, gels such as thermal gelation systems, as is well known to those skilled in the art Copolymerization systems, micelles, liposomes, microspheres, nanoparticles, liquid crystals and their dispersions, L2 phases and their dispersions, those skilled in the art are familiar with the phase behavior of oil-water systems, polymeric micelles, Multiple emulsions, self-emulsifying, in microemulsification, cyclodextrin or its derivatives, and dendrimers.

本發明的組合物適用於固體、半固體、粉末和肺部給藥用溶液製劑，例如用於按計量的計量吸入器、乾粉吸入器和噴霧器，所有本領域技術人員熟知的設備。本發明的組合物尤其適用於可控的、持續的、延長的、延遲反沖和緩慢釋放的藥物給藥系統。本發明的組合物進一步適用於，但不限於，本領域技術人員熟知的胃腸外控制釋放和持續釋放系統(兩種系統均使用藥量多倍減少)。進一步優選，皮下施用的控制釋放和持續釋放系統。下述提供的例子並不限制本發明的範圍，適用的緩控系統和組合物的實施例選自水凝膠、油性凝膠，液晶，聚合物膠束，微球，納米粒子，生產適用于本發明組合物的控制釋放系統的方法，包括但不限於，結晶法、冷凝法、工結晶法、沉澱法、共沉澱法、乳化法、分散法、高壓勻漿法、包埋法、噴霧乾燥法、微膠囊法、凝聚法、相分離法、備微球體的溶劑蒸發法制、擠出法和超臨界流體法。The compositions of the present invention are suitable for use in solution formulations for solid, semi-solid, powder and pulmonary administration, such as metered dose inhalers, dry powder inhalers and nebulizers, all of which are well known to those skilled in the art. The compositions of the present invention are particularly useful in controlled, sustained, extended, delayed recoil and slow release drug delivery systems. The compositions of the present invention are further suitable for, but not limited to, parenteral controlled release and sustained release systems well known to those skilled in the art (both systems use multiple fold reductions). Further preferred is a controlled release and sustained release system for subcutaneous administration. The examples provided below do not limit the scope of the invention, and examples of suitable retardation systems and compositions are selected from the group consisting of hydrogels, oily gels, liquid crystals, polymer micelles, microspheres, nanoparticles, and are suitable for use in production. A method of controlling the release system of the composition of the present invention, including but not limited to, crystallization, condensation, crystallization, precipitation, coprecipitation, emulsification, dispersion, high pressure homogenization, embedding, spray drying Method, microcapsule method, coacervation method, phase separation method, solvent evaporation method for preparing microspheres, extrusion method and supercritical fluid method.

通常可參考：Handbook of Pharmaceutical Controlled Release(Wise,D. L.,ed. Marcel Dekker,New York,2000)及Drug and the Pharmaceutical Sciences vol. 99: Protein Formulation and Delivery (MacNally,E.J.,ed. Marcel Dekker,New York,2000)。胃腸外給藥可採用注射器，任意的筆型注射器經皮下、肌肉內、腹膜內或靜脈內注射完成。優選地，胃腸外給藥可採用輸液泵形式完成。較優選，用於本發明化合物給藥的溶液或懸液或粉末的組合物的劑型可採用鼻用液體、肺部用液體或粉末噴劑。更優選，含有本發明化合物的藥物組合物也適用于經皮給藥，例如通過無針頭注射或藥貼，任意的電離子導入藥貼，或透粘膜給藥如口腔。本發明的化合物可使用任意已知型號的適用於肺部藥物給藥的設備，以一種載體如溶液、懸液或乾粉形式經肺部途徑給藥。這些具體實例包括，但不限於，三種通用型號的用於肺部給藥的氣溶膠發生器，和可包括噴嘴或超聲波噴霧器、可計量的劑量吸入器或乾粉吸入器(Cf. Yu J,Chien YW. Pulmonary drug delivery: Physiologic and mechanistic aspects. Crit Rev Ther Drug Carr Sys 14(4)(1997) 395-453)。Reference is generally made to: Handbook of Pharmaceutical Controlled Release (Wise, DL, ed. Marcel Dekker, New York, 2000) and Drug and the Pharmaceutical Sciences vol. 99: Protein Formulation and Delivery (MacNally, EJ, ed. Marcel Dekker, New York , 2000). For parenteral administration, a syringe can be used, and any pen-type syringe is administered subcutaneously, intramuscularly, intraperitoneally or intravenously. Preferably, parenteral administration can be accomplished in the form of an infusion pump. More preferably, the dosage form of the solution or suspension or powder composition for administration of the compound of the present invention may be a nasal liquid, a liquid for the lung or a powder spray. More preferably, the pharmaceutical compositions containing the compounds of the invention are also suitable for transdermal administration, for example by needle-free injection or patch, any iontophoresis, or transmucosal administration such as buccal. The compounds of the present invention can be administered by the pulmonary route in a carrier such as a solution, suspension or dry powder using any known type of device suitable for pulmonary drug administration. These specific examples include, but are not limited to, three general models of aerosol generators for pulmonary administration, and may include nozzles or ultrasonic nebulizers, metered dose inhalers or dry powder inhalers (Cf. Yu J, Chien YW. Pulmonary drug delivery: Physiologic and mechanistic aspects. Crit Rev Ther Drug Carr Sys 14(4) (1997) 395-453).

基於標準的測試方法學，粒子的空氣動力學直徑(d_a )是指單位密度(1g/cm³ )的參考標準的球形粒子的幾何當量直徑。在最簡單的情況下，對於球形粒子，d_a 是將參考直徑(d)作為密度比的平方根函數，其描述如下：對於聚類粒子，這種關係需要進行修正(cf. Edwards DA,Ben-Jebria A,Langer R. Recent advances in pulmonary drug delivery using large,porous inhaled particles. J Appl Physiol 84(2)(1998) 379-385)。術語“MMAD”和“MMEAD”已有詳細報導且為技術人員已知(cf. Edwards DA,Ben-Jebria A,Langer R and represents a measure of the median value of an aerodynamic particle size distribution. Recent advances in pulmonary drug delivery using large,porous inhaled particles. J Appl Physiol 84(2)(1998)379-385)。質量中值空氣動力學直徑(Mass median aerodynamic diameter,MMAD)和質量中值有效空氣動力學直徑(mass median effective aerodynamic diameter,MMEAD)可交替使用，它們是兩個統計學參數，用於經驗地描述關係氣溶膠粒子在肺部沉積潛力的尺寸因素，即獨立的實際形狀、實際大小或實際密度(cf. Edwards DA,Ben-Jebria A,Langer R. Recent advances in pulmonary drug delivery using large,porous inhaled particles. J Appl Physiol 84(2)(1998)379-385)。MMAD是通常從碰撞採樣器測量值計算，該碰撞採樣器用於測量粒子在空氣中的慣性運動。在一個優選實施方案中，所述製劑能夠被任何已知的噴霧技術霧化，如霧化，以實現氣溶膠粒子的MMAD小於10μm，特別優選1-5μm，更優選1-3μm。優選的粒子尺寸基於用於可蛋白最佳吸收的深度肺部給藥的最有效尺寸(cf. Edwards DA,Ben-Jebria A,Langer R. Recent advances in pulmonary drug delivery using large,porous inhaled particles. J Appl Physiol 84(2)(1998)379-385)。Based on the standard test methodology, the aerodynamic diameter (d _a ) of _a particle refers to the geometric equivalent diameter of a spherical particle of reference standard per unit density (1 g/cm ³ ). In the simplest case, for spherical particles, d _a is the reference root diameter (d) as a function of the square root of the density ratio, which is described as follows: For clustering particles, this relationship needs to be corrected (cf. Edwards DA, Ben- Jebria A, Langer R. Recent advances in pulmonary drug delivery using large, porous inhaled particles. J Appl Physiol 84(2) (1998) 379-385). The terms "MMAD" and "MMEAD" have been reported in detail and are known to the skilled person (cf. Edwards DA, Ben-Jebria A, Langer R and represents a measure of the median value of an aerodynamic particle size distribution. Recent advances in pulmonary) Drug delivery using large, porous inhaled particles. J Appl Physiol 84(2) (1998) 379-385). Mass median aerodynamic diameter (MMAD) and mass median effective aerodynamic diameter (MMEAD) are used interchangeably. They are two statistical parameters for empirical description. The size factor of the relationship between the deposition potential of aerosol particles in the lungs, ie the actual actual shape, the actual size or the actual density (cf. Edwards DA, Ben-Jebria A, Langer R. Recent advances in pulmonary drug delivery using large, porous inhaled particles J Appl Physiol 84 (2) (1998) 379-385). The MMAD is typically calculated from a collision sampler that measures the inertial motion of the particles in the air. In a preferred embodiment, the formulation can be atomized by any known spray technique, such as atomization, to achieve an aerosol particle having an MMAD of less than 10 μm, particularly preferably 1-5 μm, more preferably 1-3 μm. The preferred particle size is based on the most effective size for deep lung administration for optimal protein absorption (cf. Edwards DA, Ben-Jebria A, Langer R. Recent advances in pulmonary drug delivery using large, porous inhaled particles. J Appl Physiol 84 (2) (1998) 379-385).

含有本發明化合物的肺部給藥製劑的深度肺部沉積可任意地進一步採用吸入技術進行優化，例如，但不限於，慢吸氣流速(如30L/min)、屏氣呼吸和刺激計時。Deep lung deposition of a pulmonary administration formulation containing a compound of the invention can optionally be further optimized using inhalation techniques such as, but not limited to, slow inspiratory flow rates (e.g., 30 L/min), breath holding breath, and stimulation timing.

術語“被穩定的製劑”是指具有高物理穩定性、高化學穩定性或高物化穩定性的製劑。The term "stabilized formulation" refers to a formulation that has high physical stability, high chemical stability, or high physical stability.

本文中術語蛋白製劑的“物理穩定性”是指：蛋白質形成生物學惰性的趨勢；和/或蛋白質因暴露在熱-機械壓力；和/或因與諸如疏水等不穩定的介面和表面作用，形成不溶性聚集物的趨勢。含水蛋白製劑的物理穩定性可採用如下手段進行評估：在不同溫度下，將裝滿該製劑的容器(如暗盒或藥盒)在機械/物理壓力(如震動)作用下暴露不同時間後，採用目視法和/或濁度測量法進行評估。該製劑的目視法是在帶有暗背景的高度聚焦光中完成的。製劑的渾濁度可用一個視覺評分值排列的渾濁程度進行表徵，如在0-3級(製劑相對於可視得分值0不渾濁，且製劑在白光中的視覺渾濁度與視覺分值3一致)。當製劑在白光中顯示目視比濁時，對於蛋白聚集態，製劑被分類為物理性質不穩定。作為選擇，製劑的渾濁度可採用技術人員熟知的簡單濁度測量法進行評估。含水蛋白製劑的物理穩定性還可以採用蛋白光譜學試劑或蛋白構象形態探針進行評估。上述探針優選能夠優先與蛋白的非天然構象異構體結合的小分子。一個蛋白結構小分子光譜學探針的實例是硫代黃素T(Thioflavin T)，一種廣泛用於類澱粉樣纖維檢測的螢光染料。當存在小纖維以及可能的其他蛋白結構時，硫代黃素T與小纖維蛋白形式結合，增加了在約450nm處的新激發最大值，增強了約482nm處的發射峰。未被結合的硫代黃素T在上述波長處本質上無螢光激發或發射。The term "physical stability" of a protein preparation herein means: a tendency for the protein to form a biologically inert; and/or the protein is exposed to thermo-mechanical pressure; and/or due to an unstable interface and surface interaction such as hydrophobicity, The tendency to form insoluble aggregates. The physical stability of the aqueous protein preparation can be evaluated by using a container filled with the preparation (such as a cassette or a kit) at different temperatures for a different period of time under mechanical/physical pressure (such as vibration). Visual and/or turbidity measurements are evaluated. The visual inspection of the formulation was done in highly focused light with a dark background. The turbidity of the formulation can be characterized by the degree of turbidity of a visual score, as in the 0-3 grade (the formulation is not turbid relative to the visual score of 0, and the visual turbidity of the formulation in white light is consistent with the visual score of 3) . When the formulation showed visual turbidity in white light, the formulation was classified as physically unstable for protein aggregation. Alternatively, the turbidity of the formulation can be assessed using simple turbidity measurements well known to the skilled artisan. The physical stability of aqueous protein preparations can also be assessed using protein spectroscopy reagents or protein conformational morphological probes. The above probes are preferably small molecules which are capable of preferentially binding to the non-native conformer of the protein. An example of a protein structure small molecule spectroscopy probe is Thioflavin T, a fluorescent dye widely used for amyloid-like fiber detection. When small fibers and possibly other protein structures are present, thioflavin T binds to the fibrin form, increasing the new excitation maximum at about 450 nm, enhancing the emission peak at about 482 nm. The unbound thioflavin T is substantially free of fluorescent excitation or emission at the above wavelengths.

其他小分子也可以作為探針，用於將蛋白結構從天然構象變為非天然構象。例如，“疏水區”螢光探針優先結合到暴露的蛋白疏水區。蛋白的疏水區通常包埋在其天然構象的三級結構中，但是當蛋白開始打開或變性時，蛋白的疏水區域會暴露出來。這些小分子、光譜探針的實例都是芳香族的疏水染料，例如蒽、吖啶、鄰二氮菲或它們的類似物。其他光譜學探針是金屬-氨基酸配合物，例如諸如苯丙氨酸、亮氨酸、異亮氨酸、蛋氨酸、纈氨酸等類似疏水氨基酸的鈷金屬配合物。Other small molecules can also be used as probes to transform the protein structure from a native conformation to a non-native conformation. For example, a "hydrophobic region" fluorescent probe preferentially binds to an exposed hydrophobic region of the protein. The hydrophobic region of a protein is usually embedded in the tertiary structure of its native conformation, but when the protein begins to open or denature, the hydrophobic region of the protein is exposed. Examples of such small molecules, spectral probes are aromatic hydrophobic dyes such as hydrazine, acridine, phenanthroline or their analogs. Other spectroscopy probes are metal-amino acid complexes such as cobalt metal complexes such as phenylalanine, leucine, isoleucine, methionine, valine and the like hydrophobic amino acids.

本文中蛋白製劑的“化學穩定性”術語是指蛋白結構中共價化學鍵的改變，生成較天然蛋白結構具有潛在低生物活性和/或潛在產生免疫系統的化學降解產物。各種化學降解產物的形成依賴于天然蛋白的類型和性質以及蛋白暴露的環境。化學降解物的消除是不可能完全避免的，化學降解產物量的增加在存儲和本領域技術人員熟知的蛋白製劑的使用過程中是常見的。大多數蛋白傾向於脫氨基過程，即谷氨醯胺醯殘基或天冬醯胺醯殘基的側鏈氨基被水解，形成游離羧酸。其他降解途徑還包括高分子量轉化產物的形成，即在轉化過程中兩個或兩個以上蛋白分子之間彼此共價結合，和/或二硫化作用導致共價結合的二聚物、低聚物和高分子降解產物的形成(Stability of Protein Pharmaceuticals,Ahem. T.J. & Manning M. C,Plenum Press,New York 1992)。此外，氧化(比如蛋氨酸殘基的氧化)也是被提到的其他化學降解途徑。蛋白製劑化學穩定性的評估方法，是將蛋白製劑暴露於不同環境條件(降解產物的形成通常與諸如升高溫度等環境條件成正比)下，測量不同時間點的化學降解產物含量。每個單個降解產物的含量通常取決於採用不同色譜設備(如SEC-HPLC和/或RP-HPLC)根據分子大小和/或電荷降解產物的分離方法。The term "chemical stability" of a protein preparation herein refers to a change in a covalent chemical bond in a protein structure that produces a chemical degradation product that has a potentially low biological activity and/or potentially produces an immune system that is greater than the native protein structure. The formation of various chemical degradation products depends on the type and nature of the native protein and the environment in which the protein is exposed. Elimination of chemical degradation products is not completely avoidable, and an increase in the amount of chemical degradation products is common during storage and use of protein preparations well known to those skilled in the art. Most proteins tend to undergo a deamination process, i.e., the glutamine azide residue or the side chain amino group of the aspartamide residue is hydrolyzed to form a free carboxylic acid. Other degradation pathways also include the formation of high molecular weight conversion products, ie, covalent bonding of two or more protein molecules to each other during conversion, and/or disulfide resulting in covalently bound dimers, oligomers And formation of polymeric degradation products (Stability of Protein Pharmaceuticals, Ahem. TJ & Manning M. C, Plenum Press, New York 1992). In addition, oxidation (such as oxidation of methionine residues) is also mentioned as another chemical degradation pathway. The chemical stability of protein preparations is evaluated by exposing the protein preparation to different environmental conditions (the formation of degradation products is usually proportional to environmental conditions such as elevated temperatures) and measuring the chemical degradation product content at different time points. The amount of each individual degradation product typically depends on the separation method based on molecular size and/or charge degradation products using different chromatographic equipment (eg, SEC-HPLC and/or RP-HPLC).

因此，如上文概述，“穩定的製劑”是指具有高物理穩定性、高化學穩定性製劑。總的說來，在到有效期之前，製劑在使用和存儲中(按照推薦的使用方和存儲條件)必須是穩定的。Thus, as outlined above, a "stable formulation" refers to a formulation that has high physical stability and high chemical stability. In general, the formulation must be stable in use and storage (according to recommended consumer and storage conditions) until the expiration date.

在本發明的一個實施方案中，所述含有本發明化合物的藥物組合物，在使用6個星期以上和存儲3年都是穩定的。In one embodiment of the invention, the pharmaceutical composition comprising a compound of the invention is stable for more than 6 weeks and for 3 years of storage.

在本發明的另一個實施方案中，所述含有本發明化合物的藥物組合物，在使用4個星期以上和存儲3年都是穩定的。在本發明的又一個實施方案中，所述含有本發明化合物的藥物組合物，在使用4個星期以上和存儲2年都是穩定的。In another embodiment of the invention, the pharmaceutical composition comprising a compound of the invention is stable for more than 4 weeks and for 3 years of storage. In still another embodiment of the invention, the pharmaceutical composition comprising a compound of the invention is stable for more than 4 weeks and for 2 years of storage.

在本發明的還一個實施方案中，所述含有本發明化合物的藥物組合物，在使用2個星期以上和存儲2年都是穩定的。In still another embodiment of the present invention, the pharmaceutical composition containing the compound of the present invention is stable for 2 weeks or more and for 2 years of storage.

本發明的另一個方面涉及根據本發明藥物製備的化合物的用途。Another aspect of the invention relates to the use of a compound prepared according to the medicament of the invention.

本發明還涉及GLP-1類似物的鹽。本發明的GLP-1類似物可以是足夠的酸性或足夠的鹼性，從而與任意當量的無機鹼或無機酸反應成鹽。通用的形成酸加成鹽的酸是：無機酸，例如鹽酸、氫溴酸、氫碘酸、硫酸、磷酸和類似的無機酸；有機酸，例如間-苯甲磺酸、甲磺酸、草酸、間-溴苯基-磺酸、碳酸、檸檬酸、苯甲酸、醋酸和類似的有機酸。上述鹽的實例包括硫酸鹽、硫酸氫鹽、重硫酸鹽、亞硫酸鹽、磷酸鹽、磷酸一氫鹽、磷酸二氫鹽、偏磷酸鹽、焦磷酸鹽、鹽酸鹽、溴酸鹽、碘酸鹽、丙酸鹽、癸酸鹽辛酸鹽、丙烯酸鹽、甲酸鹽、異丁酸鹽、庚酸鹽、丙炔酸鹽、草酸鹽、丙二酸鹽、琥珀酸鹽、辛二酸鹽、癸二酸鹽、延胡索酸鹽、馬來酸鹽、丁炔-1,4-二酸鹽、己炔-1,6-二酸鹽、本甲酸鹽、氯苯甲酸鹽、甲基本甲酸鹽、二硝基苯甲酸鹽、羥基苯甲酸鹽、甲氧基本甲酸鹽、鄰苯二甲酸鹽、磺酸鹽、二甲苯磺酸鹽、苯基丙酸鹽、苯基丁酸鹽、檸檬酸鹽、乳酸鹽、γ-羥基丁酸鹽、羥乙酸鹽、酒石酸鹽、甲基磺酸鹽、丙基磺酸鹽、萘基-1-磺酸鹽、萘基-2-磺酸鹽、扁桃酸鹽和類似的酸加成鹽。優選的酸加成鹽，是那些與無機酸形成的鹽，例如鹽酸、氫溴酸或氫氟酸，特別是鹽酸。The invention also relates to salts of GLP-1 analogs. The GLP-1 analogs of the invention may be sufficiently acidic or sufficiently basic to react with any equivalent of an inorganic or inorganic acid to form a salt. Commonly used acids which form acid addition salts are: inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid and similar inorganic acids; organic acids such as m-benzenesulfonic acid, methanesulfonic acid, oxalic acid , m-bromophenyl-sulfonic acid, carbonic acid, citric acid, benzoic acid, acetic acid and similar organic acids. Examples of the above salts include sulfates, hydrogen sulfates, heavy sulfates, sulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, hydrochlorides, bromates, and iodine. Acid salt, propionate, citrate octanoate, acrylate, formate, isobutyrate, heptanoate, propiolate, oxalate, malonate, succinate, suberic acid Salt, sebacate, fumarate, maleate, butyne-1,4-diate, hexyne-1,6-diate, present formate, chlorobenzoate, methyl Formate, dinitrobenzoate, hydroxybenzoate, methoxy carboxate, phthalate, sulfonate, xylene sulfonate, phenylpropionate, phenyl Butyrate, citrate, lactate, γ-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propyl sulfonate, naphthyl-1-sulfonate, naphthyl-2 a sulfonate, a mandelate and a similar acid addition salt. Preferred acid addition salts are those formed with mineral acids such as hydrochloric acid, hydrobromic acid or hydrofluoric acid, especially hydrochloric acid.

鹼加成鹽包括那些來源於無機鹼，例如銨鹽或鹼金屬氫氧化物或鹼土金屬氫氧化物、碳酸鹽、碳酸氫鹽和類似的無機鹼。因此，這些用於製備本發明鹽的鹼包括氫氧化鈉、氫氧化鉀、氨水、碳酸鉀和它們類似的鹼。特別優選GLP-1類似物的鹽形式。當然，當本發明的化合物被用於治療目的時，這些化合物也可以是鹽形式，但是這些鹽必須是藥學上可接受的。Base addition salts include those derived from inorganic bases such as ammonium or alkali metal hydroxides or alkaline earth metal hydroxides, carbonates, hydrogencarbonates and similar inorganic bases. Accordingly, these bases for the preparation of the salts of the present invention include sodium hydroxide, potassium hydroxide, aqueous ammonia, potassium carbonate and similar bases thereof. Salt forms of the GLP-1 analog are particularly preferred. Of course, when the compounds of the invention are used for therapeutic purposes, these compounds may also be in the form of a salt, but these salts must be pharmaceutically acceptable.

發明人發現本發明的經修飾的GLP-1類似物具有多重用途，包括用作糖尿病治療方法、鎮靜劑、用作神經系統基本疾病治療方法、用於誘導對於中樞神經系統(CNS)的抗焦慮效果、用於刺激中樞神經系統、用於術後治療和用作胰島素抵抗的治療方法。The inventors have found that the modified GLP-1 analogs of the present invention have versatile uses, including use as a method of treating diabetes, a sedative, as a treatment for essential diseases of the nervous system, for inducing an anxiolytic effect on the central nervous system (CNS). For the stimulation of the central nervous system, for postoperative treatment and as a treatment for insulin resistance.

A.糖尿病治療A. Diabetes treatment

本發明的經修飾的GLP-1類似物普遍能通過葡萄糖依賴機制使高血糖症正常化。類似地，該經修飾的GLP-1類似物可作為治療II型糖尿病的主要試劑和治療I型糖尿病的輔助試劑。The modified GLP-1 analogs of the invention generally normalize hyperglycemia by a glucose-dependent mechanism. Similarly, the modified GLP-1 analog can be used as a primary agent for the treatment of type 2 diabetes and as an adjunct to the treatment of type 1 diabetes.

使用有效劑量的經修飾的GLP-1類似物用於治療糖尿病的優勢在於：比未經修飾的GLP-1類似物更有效。由於經修飾的GLP-1類似物在體內的更穩定，因此服用更小劑量的分子可達到有效的治療效果。本發明尤其適用于治療糖尿病患者，包括I型和II型糖尿病。在糖尿病患者的治療中，GLP-1類似物多肽的作用是依賴於血液中血糖的濃度，因此在使用現有治療方法上大大地降低了低血糖症副作用的風險。An advantage of using an effective amount of a modified GLP-1 analog for treating diabetes is that it is more effective than an unmodified GLP-1 analog. Since the modified GLP-1 analog is more stable in vivo, a smaller dose of the molecule can achieve an effective therapeutic effect. The invention is particularly useful for treating diabetic patients, including type I and type II diabetes. In the treatment of diabetic patients, the effect of the GLP-1 analog polypeptide is dependent on the concentration of blood glucose in the blood, thus greatly reducing the risk of side effects of hypoglycemia using existing therapies.

本發明還提供了一種治療個體糖尿病的方法，其中所述方法包括提供至少一種一定劑量的足以治療糖尿病的經修飾的GLP-1類似物；所述組合物含有至少一種經修飾的GLP-1類似物。The invention also provides a method of treating diabetes in a subject, wherein the method comprises providing at least one dose of a modified GLP-1 analogue sufficient to treat diabetes; the composition comprising at least one modified GLP-1 similar Things.

B.　神經系統紊亂的治療B. Treatment of nervous system disorders

發明人發現本發明的經修飾的GLP-1類似物可用作鎮靜劑。在本發明的一個方面，本發明再提供一種鎮靜哺乳動物物件的方法：對於一些因某種異常而導致中樞或外周神經系統的活性被增強的哺乳動物，使用足夠劑量的經修飾的GLP-1類似物，可已在這些哺乳動物身上產生一種誘導的或抗焦慮效果，從而使其鎮靜。這些經修飾的GLP-1類似物可經腦室內注射、口服、皮下注射、肌肉內注射或靜脈注射。該方法可用於治療或改善神經系統症候，例如焦慮、運動錯亂、侵略性行為、精神錯亂、癲癇、驚恐發作、歇斯底里病和睡眠障礙等神經系統症候。The inventors have found that the modified GLP-1 analogs of the invention are useful as sedatives. In one aspect of the invention, the invention further provides a method of sedating a mammalian article: for a mammal having increased activity of the central or peripheral nervous system due to an abnormality, a sufficient dose of modified GLP-1 is used Analogs, which may have produced an induced or anxiolytic effect in these mammals, thereby calming them. These modified GLP-1 analogs can be administered intracerebroventricularly, orally, subcutaneously, intramuscularly, or intravenously. The method can be used to treat or ameliorate nervous system symptoms such as anxiety, movement disorders, aggressive behavior, confusion, epilepsy, panic attacks, hysteria and sleep disorders.

在另一方面，本發明包含一種提高哺乳動物對象活動性的方法，包括給予對象做夠劑量經修飾的GLP-1類似物，使其在哺乳動物身上產生並激活對象的功效。優選地，所述對象具有中樞神經系統或周圍神經系統活性降低特點。發明人發現經修飾的GLP-1類似物尤其適用於治療或改善抑鬱症、***情感性障礙、睡眠呼吸中止症、注意力分散症、失憶症、健忘症、嗜睡症等一些列症狀。在這些病症中，喚醒中樞神經系統可能是有利的。In another aspect, the invention comprises a method of increasing the activity of a mammalian subject comprising administering to the subject a dose of a modified GLP-1 analogue to produce and activate the subject in a mammal. Preferably, the subject has a central nervous system or peripheral nervous system activity reduction profile. The inventors have found that modified GLP-1 analogs are particularly useful for treating or ameliorating a number of symptoms such as depression, schizoaffective disorder, sleep apnea, attention distraction, amnesia, amnesia, narcolepsy and the like. Among these conditions, awakening the central nervous system may be advantageous.

本發明經修飾的GLP-1類似物也可用於誘導激發，從而用於治療或緩解抑鬱症、***情感性障礙、睡眠呼吸中止症、注意力分散症、失憶症、健忘症、嗜睡症。經修飾的GLP-1類似物療法的治療效果可通過患者會診來評估他們的病症，通過心理學上或神經學實驗，或通過與這些病症相關症狀的改善來進行評定。例如，從對嗜睡症發作的掌控程度可以來評價其對嗜睡症的治療效果。再例如，可用本領域技術人員熟知的各種不同診斷測試中的任何一種，來評估經修飾的GLP-1類似物作用于患者後使之集中注意力或增強記憶力的效果。The modified GLP-1 analogs of the invention can also be used to induce challenge for the treatment or alleviation of depression, schizoaffective disorder, sleep apnea, attention distraction, amnesia, amnesia, narcolepsy. The therapeutic effects of modified GLP-1 analog therapies can be assessed by patient consultations, assessed by psychological or neurological experiments, or by improvements in symptoms associated with these conditions. For example, the therapeutic effect on narcolepsy can be evaluated from the degree of control over the onset of narcolepsy. As another example, any of a variety of different diagnostic tests well known to those skilled in the art can be used to assess the effect of the modified GLP-1 analog upon focusing on the patient or enhancing memory.

C.術後治療C. Postoperative treatment

本發明的經修飾的GLP-1類似物可用於術後治療。無論在術前1-16小時，或手術過程中，或者術後5天內，患者都是需要本發明的經修飾的GLP-1類似物。The modified GLP-1 analogs of the invention are useful for post-operative treatment. The patient is in need of the modified GLP-1 analog of the invention, either 1-16 hours before surgery, or during surgery, or within 5 days of surgery.

從在手術開始之前的16小時至前1小時，對患者施用經修飾的GLP-1類似物。給予患者本發明的化合物，以減少分解代謝作用和胰島素耐受性，而給藥時間的長短取決於許多因素。這些因素是為醫師所常規熟知的，包括，最重要的手術前準備階段，患者是否禁食或進行了葡萄糖輸注或引用了飲料或者其他形式的食物。其他重要因素包括患者的性別、體重、年齡、血糖的嚴重性、任何調節血糖無能的潛在誘因，手術預期創傷的嚴重性、給藥途徑以及生物利用度、個體耐受性、處方和藥效。開始給予本發明使用的經修飾的GLP-1類似物的優選時間是術前大約1小時到大約10小時。進一步優選的開始給予時間是術前2小時到8小時。The modified GLP-1 analog was administered to the patient from 16 hours to 1 hour prior to the start of the procedure. The compounds of the invention are administered to a patient to reduce catabolism and insulin resistance, and the length of administration depends on a number of factors. These factors are routinely known to physicians, including, during the most important pre-operative preparation phase, whether the patient is fasting or has undergone glucose infusion or cites a drink or other form of food. Other important factors include the patient's gender, weight, age, severity of blood glucose, any potential cause of regulating blood sugar incompetence, severity of surgically anticipated trauma, route of administration, and bioavailability, individual tolerance, prescription, and efficacy. The preferred time to start administering the modified GLP-1 analog used in the present invention is from about 1 hour to about 10 hours before surgery. A further preferred start administration time is 2 hours to 8 hours before surgery.

在特殊類型手術後，例如選擇性腹部手術後的的胰島素抵抗在手術後的第一天最為嚴重，持續至少5天，並且可能持續三周才能恢復正常。因此，經歷了手術創傷，術後一段時間，患者需要施用本發明的經修飾的GLP-1類似物，給藥時間的長短取決於患者術後是否禁食或者進行了葡萄糖輸注或飲用了飲料或其他形式的食物。同時，不限於，患者的性別、體重和年齡、血糖失調的嚴重性、能引起血糖調節功能喪失的潛在誘因、手術創傷的實際嚴重性、給藥途徑、生物利用度、個體耐受度、處方和藥效也是影響因素。本發明使用的化合物的優選給藥持續時間為不多於術後五天。After a particular type of surgery, for example, insulin resistance after selective abdominal surgery is most severe on the first day after surgery, lasting at least 5 days, and may last for three weeks to return to normal. Therefore, after undergoing surgical trauma, the patient needs to administer the modified GLP-1 analogue of the present invention for a period of time after surgery, depending on whether the patient is fasted or has undergone glucose infusion or drinking a drink or Other forms of food. At the same time, not limited to, the patient's gender, weight and age, the severity of blood glucose disorders, potential causes of loss of blood glucose regulation, the actual severity of surgical trauma, route of administration, bioavailability, individual tolerance, prescription And efficacy is also a factor. The preferred duration of administration of the compounds used in the present invention is no more than five days after surgery.

D.　胰島素耐受性治療D. Insulin resistance treatment

本發明經修飾的GLP-1類似物，可用於治療胰島素抵抗，無論其是否用於術後治療中。胰島素抵抗的原因可能是細胞表面受體結合的胰島素數量減少，或細胞內代謝的改變。第一種胰島素抵抗的原因的特點是胰島素敏感性降低，可通過增加胰島素濃度克服。第二種胰島素抵抗的原因的特點是胰島素反應性降低，通過大劑量胰島素無效。胰島素抵抗繼發的創傷可通過與胰島素抵抗成比例的增加胰島素劑量克服，因此這種創傷很明顯地是由胰島素敏感性降低引起的。The modified GLP-1 analogs of the invention are useful for treating insulin resistance, whether or not they are used in post-operative treatment. The cause of insulin resistance may be a decrease in the amount of insulin bound by cell surface receptors, or a change in intracellular metabolism. The first cause of insulin resistance is characterized by a decrease in insulin sensitivity, which can be overcome by increasing the insulin concentration. The second cause of insulin resistance is characterized by a decrease in insulin reactivity, which is ineffective by high doses of insulin. The trauma secondary to insulin resistance can be overcome by increasing the insulin dose in proportion to insulin resistance, so this trauma is clearly caused by a decrease in insulin sensitivity.

使患者血糖水準正常化所用本發明經修飾的GLP-1類似物的劑量受以下因素的影響：程度包括但不僅限於，患者性別、體重和年齡、血糖調節功能喪失的嚴重程度、能引起血糖調節功能喪失的潛在病因、是否同時給予葡萄糖或其他糖類、給藥途徑及生物利用度、患者耐受度、處方和藥效。The dose of the modified GLP-1 analogue of the invention used to normalize the blood glucose level of a patient is affected by factors including, but not limited to, the patient's gender, weight and age, the severity of loss of glycemic regulation, and the ability to cause glycemic regulation. The underlying cause of loss of function, whether glucose or other sugars are administered simultaneously, the route of administration and bioavailability, patient tolerance, prescription, and efficacy.

要檢測GLP-1類似物刺激胰島素分泌的能力，可以將GLP-1類似物注入人工培養的動物細胞中，例如RIN-38型小數胰島β細胞瘤細胞，然後監測釋放入人工配基基中的免疫反應性胰島素(immunoreactive insulin,IRI)的數量。另一種方法是將GLP-1類似物注入動物體內，然後監測免疫反應胰島素(immunoreactive insulin,IRI)在血漿中的濃度。To detect the ability of GLP-1 analogs to stimulate insulin secretion, GLP-1 analogs can be injected into artificially cultured animal cells, such as RIN-38 fractional islet beta cell tumor cells, and then monitored for release into the artificial ligand base. The number of immunoreactive insulin (IRI). Another method is to inject the GLP-1 analogue into the animal and then monitor the concentration of immunoreactive insulin (IRI) in the plasma.

IRI的存在是用針對胰島素放射性免疫測定法檢測的。任何能檢測到IRI的放射性免疫化驗方法都可以使用，其中一種是由Albano,J.D.M.等發明的改良方法(Albano,J.D.M. et al.,Acta Endocrinol. 1972,70: 487-509)。在這種改良方法中，使用pH7.4的磷酸鹽/白蛋白緩衝液。向培養液依次加入500μL磷酸緩衝液、50μL灌注液樣本或含有大鼠胰島素標準量的灌注液、100μL抗胰島素抗血清(Wellcome Laboratories：1：40,000稀釋)和100μL[¹²⁵ I]胰島素，將總體積750μL置於10×75mm一次性玻璃試管中。在4℃下培育2-3天，用活性炭分離游離的胰島素和抗體結合的胰島素。該化驗方法的靈敏度為1-2uU/ml。為了測定釋放到組織培養物中細胞生長基德IRI數量，可以給胰島素原加上放射性標記。儘管任何能標記的放射性標記都可使用，但優選使用³ H亮氨酸來獲得被標記的胰島素原。The presence of IRI is detected by a radioimmunoassay for insulin. Any radioimmunoassay method capable of detecting IRI can be used, one of which is an improved method invented by Albano, JDM, etc. (Albano, JDM et al., Acta Endocrinol. 1972, 70: 487-509). In this modified method, a phosphate/albumin buffer of pH 7.4 is used. 500 μL of phosphate buffer, 50 μL of perfusate sample or a perfusion solution containing rat insulin standard amount, 100 μL of anti-insulin antiserum (Wellcome Laboratories: 1:40,000 dilution) and 100 μL of [ ¹²⁵ I] insulin were added to the culture solution in turn, and the total volume was added. 750 μL was placed in a 10 x 75 mm disposable glass test tube. Incubate at 4 ° C for 2-3 days to separate free insulin and antibody-bound insulin with activated carbon. The sensitivity of this assay is 1-2 uU/ml. To determine the amount of cell growth Kidide IRI released into tissue culture, the proinsulin can be radiolabeled. Although any radiolabel that can be labeled can be used, it is preferred to use ³ H leucine to obtain the labeled proinsulin.

要測定GLP-1類似物是否有促胰島功能也可以通過胰腺灌注實驗。大鼠胰腺原位灌注試驗是在Penhos,J,C.等人的方法基礎上的改良方法(Penhos,J,C.,et al.,Diabetes,1969,18:733-738)。在體重350g-600g不等的、禁食的雄性Charles River種大鼠胰腺腹膜內注射異戊巴比妥(Eli Lilly and Co,160ng/kg)，將它們麻醉，結紮腎臟、腎上腺、胃及低位結腸的血管。除了大約4釐米的十二指腸和降結腸以及直腸，整段腸都失去作用功能。因此，只有一小段腸受到灌注，從而最大程度地減少了胰高血糖素樣肽與腸物質之間免疫反應所造成的影響。灌注液是改良後的Kreba-Ringer碳酸氫鹽緩衝液，含有4%的T70右旋糖酐和0.2%的牛血清白蛋白(片段V)，還用95%的O₂ 和5%的CO₂ 鼓泡。使用了一台帶4通道滾珠軸承泵的體外迴圈機(Buchler polystatic,Buchler Instruments Division,Nuclear-Chicago Corp)，還有一個控制灌注液通向的三通閥。該灌注法參見Weir,G. C.等人的方法進行操作、監測和分析(Weir,G. C.,et al.,. J. Clin. Investigat. 1974,54:1403-1412)。To determine if GLP-1 analogues have islet promoting function, pancreatic perfusion experiments can also be performed. The rat pancreatic in situ perfusion test is an improved method based on the method of Penhos, J, C. et al. (Penhos, J, C., et al., Diabetes, 1969, 18: 733-738). In the pancreas of fasting male Charles River rats weighing 350g-600g, intraperitoneal injection of pentobarbital (Eli Lilly and Co, 160ng/kg), anesthesia, ligation of kidney, adrenal gland, stomach and low position The blood vessels of the colon. Except for the duodenum and descending colon and the rectum, which are about 4 cm, the entire intestine loses its function. Therefore, only a small portion of the intestine is perfused, thereby minimizing the effects of the immune response between the glucagon-like peptide and the intestinal material. The perfusate was a modified Kreba-Ringer bicarbonate buffer containing 4% T70 dextran and 0.2% bovine serum albumin (fragment V), also bubbled with 95% O ₂ and 5% CO ₂ . An extracorporeal looper with a 4-channel ball bearing pump (Buchler polystatic, Buchler Instruments Division, Nuclear-Chicago Corp) was used, as well as a three-way valve that controls the flow of perfusate. The perfusion method is operated, monitored and analyzed by the method of Weir, GC et al. (Weir, GC, et al., J. Clin. Investigat. 1974, 54: 1403-1412).

使用本發明化合物的治療方法也可與第二種或更多藥學上接受的物質聯合，例如，選自抗糖尿病藥、抗肥胖藥、食欲調節藥、抗高血壓藥、治療和/或糖尿病導致的或伴隨的綜合症的藥物、治療和/或預防肥胖導致的或伴隨的綜合症的藥物。這些藥物活性物質的具體實例是：胰島素、磺脲類藥物、雙胍類藥物、氯茴苯酸類藥物、普糖苷酶抑制劑、胰高血糖素拮抗劑、高效二肽基肽酶-IV(DPP-IV)抑制劑、葡質異生和/或糖原病刺激相關的肝損傷酶的抑制劑、葡萄糖吸收調節劑、修飾脂質代謝的化合物例如抗高血脂藥如HMG CoA抑制劑(他汀類藥物)、胃泌素抑制肽(Gastric Inhibitory Polypeptides,GIP類似物)、降低食物攝入的化合物、RXR激動劑和作用域ATP-依賴的β-細胞的鉀通道藥物；考來烯胺(Cholestyramine)、考來替泊(colestipol)、氯貝丁酯(clofibrate)、吉非羅齊(gemfibrozil)、洛伐他汀(lovastatin)、普伐他汀(pravastatin)、辛伐他汀(simvastatin)、普羅布考(probucol)、右旋甲狀腺素(dextrothyroxine)、那格列奈(neteglinide)、瑞格列奈(repaglinide)；β-阻滯劑，例如阿普洛爾(alprenolol)、阿替洛爾(atenolol)、噻嗎洛爾(timolol)、吲哚洛爾(pindolol)、普萊洛爾(propranolol)和美托洛爾(metoprolol)、血管緊張肽酶轉化酶(angiotensin converting enzyme,ACE)抑制劑，例如貝那普利(benazepril)、卡托普利(captopril)、依拉普利(enalapril)、福辛普利(fosinopril)、賴羅普利(lisinopril)、alatriopril、喹那普利(quinapril)和雷米普利(ramipril)、鈣通道阻滯劑，例如硝苯地平(nifedipine)、非洛地平(felodipine)、尼卡地平(nicardipine)、伊拉地平(isradipine)、尼莫地平(nimodipine)、地爾硫卓(diltiazem)和異搏定(verapamil)和α-阻滯劑例如多沙唑嗪(doxazosin)、烏拉地爾(urapidil)、呱唑嗪(prazosin)和特拉唑嗪(terazosin)；***-***調節轉錄肽(***e amphetamine regulated transcript,CART)激動劑、神經肽Y(neuropeptide Y,NPY)拮抗劑、PYY激動劑PYY2激動劑、PYY4激動劑、混合的PPY2/PYY4激動劑、黑素腎上腺皮質激素4(melanocortin 4,MC4)激動劑、促食素拮抗劑、腫瘤壞死因子(tumor necrosis factor,TNF)激動劑、促腎上腺皮質激素釋放因子(corticotropin releasing factor,CRF)激動劑、促腎上腺皮質激素釋放因子結合蛋白(corticotropin releasing factor binding protein,CRF BP)拮抗劑、尿皮質素激動劑、β3激動劑、黑素細胞刺激素(melanocyte-stimulating hormone,MSH)激動劑、黑色素濃集激素(melanocyte-concentrating hormone,MCH)拮抗劑、腸促胰酶肽(cholecystokinin,CCK)激動劑、血清素再吸收抑制劑、血清素和去腎上腺素再吸收抑制劑、混合的血清素和去甲腎上腺素啟動的化合物、血清素(serotonin,5HT)激動劑、鈴蟾肽激動劑、甘丙肽拮抗劑、生長素、生長素釋放化合物、促甲狀素釋放素(thyreotropin releasing hormone,TRH)激動劑、UCP 2解偶聯蛋白(uncoupling protein,UCP)或UCP 3調節劑、leptin激動劑、DA激動劑(溴麥角環肽(bromocriptin),doprexin)、脂肪酶/澱粉酶抑制劑、類視黃醇X受體(retinoid X receptor,RXR)調節劑、TR β激動劑；組氨酸H3拮抗劑、胃泌素抑制肽激動劑或拮抗劑(Gastric Inhibitory Polypeptide類似物,GIP類似物)、胃泌素(gastrin)和胃泌素類似物。使用本發明化合物的治療方法，也可以聯合影響葡萄糖水準和/或體脂平衡的外科手術-外科手術，例如束胃帶手術或胃繞道手術。The method of treatment using a compound of the invention may also be combined with a second or more pharmaceutically acceptable substance, for example, selected from the group consisting of an anti-diabetic agent, an anti-obesity agent, an appetite regulating drug, an antihypertensive drug, a therapeutic and/or a diabetic Or a concomitant syndrome drug, a drug that treats and/or prevents obesity or the accompanying syndrome. Specific examples of such pharmaceutically active substances are: insulin, sulfonylureas, biguanides, meglitinides, glucosidase inhibitors, glucagon antagonists, high-efficiency dipeptidyl peptidase-IV (DPP- IV) Inhibitors, inhibitors of liver damage enzymes associated with gluconeogenesis and/or glycogen stimulation, glucose uptake modulators, compounds that modify lipid metabolism such as antihyperlipidemics such as HMG CoA inhibitors (statins) , Gastric Inhibitory Polypeptides (GIP Analogs), Compounds that Reduce Food Intake, RXR Agonists, and ATP-Dependent β-Cell Potassium Channel Drugs; Cholestyramine Colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol , dextrothyroxine, neteglinide, repaglinide; beta-blockers, such as aprenolol, atenolol, thiophene Timolol, pindolol , propranolol and metoprolol, angiotensin converting enzyme (ACE) inhibitors, such as benazepril, captopril, Elapaliril, fosinopril, lisinopril, alatriopril, quinapril and ramipril, calcium channel blockers such as nifedipine Nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil and alpha-blockers For example, doxazosin, urapidil, prazosin, and terazosin; ***e amphetamine regulated transcript (CART) agonists, neuropeptides Y (neuropeptide Y, NPY) antagonist, PYY agonist PYY2 agonist, PYY4 agonist, mixed PPY2/PYY4 agonist, melanometric corticosteroid 4 (MC4) agonist, progesterone antagonist, Tumor necrosis factor (TNF) An agonist, a corticotropin releasing factor (CRF) agonist, a corticotropin releasing factor binding protein (CRF BP) antagonist, a urocortin agonist, a β3 agonist, Melanocyte-stimulating hormone (MSH) agonist, melanocyte-concentrating hormone (MCH) antagonist, cholestyrintokinin (CCK) agonist, serotonin reuptake inhibitor , serotonin and norepinephrine reuptake inhibitors, mixed serotonin and norepinephrine-initiated compounds, serotonin (5HT) agonists, bombesin agonists, galanin antagonists, auxin, Auxin releasing compound, thyreotropin releasing hormone (TRH) agonist, UCP 2 uncoupling protein (UCP) or UCP 3 modulator, leptin agonist, DA agonist (bromo ergot Bromocriptin, doprexin), lipase/amylase inhibitor, retinoid X receptor (RXR) modulator, TR beta agonist Histidine H3 antagonists, gastrin inhibitory peptide agonists or antagonists (Gastric Inhibitory Polypeptide analogs, the GIP analogs), gastrin (Gastrin) and gastrin analogs. Treatments using the compounds of the invention may also be combined with surgical-surgical procedures that affect glucose levels and/or body fat balance, such as gastric banding surgery or gastric bypass surgery.

可以理解任何適合的本發明的化合物聯合一種或一種以上上述化合物和任意地一種或一種以上藥學上用的活性物質，也被考慮在本發明的保護範圍內。It is to be understood that any suitable compound of the invention in combination with one or more of the above compounds and optionally one or more pharmaceutically active substances is also contemplated as being within the scope of the invention.

為使　貴審查委員能進一步瞭解本發明之結構，特徵及其他目的，玆以如後之較佳實施例附以圖式詳細說明如後，惟本圖例所說明之實施例係供說明之用，並非為專利申請上之唯一限制者。The structure, features and other objects of the present invention will be further described in the following detailed description of the preferred embodiments of the present invention. It is not the only restriction on patent applications.

具體實施方式detailed description

縮寫對照表Abbreviation comparison table

r.t：室溫；R.t: room temperature;

H₂ O：水；H ₂ O: water;

CH₃ CN：乙腈；CH ₃ CN: acetonitrile;

DMF：N,N-二甲基甲醯胺；DMF: N,N-dimethylformamide;

HBTU：O-(1H-苯並三氮唑-1-基)-1,1,3,3-四甲基脲六氟磷酸鹽；HBTU: O-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea hexafluorophosphate;

Fmoc：9-芴甲氧羰基；Fmoc: 9-fluorenylmethoxycarbonyl;

Boc：叔丁氧羰基；Boc: tert-butoxycarbonyl;

OtBu：叔丁基酯；OtBu: tert-butyl ester;

tBu：叔丁基；tBu: tert-butyl;

Trt：三苯甲基；Trt: trityl;

Pmc：2,2.5,7,8-五甲基苯並二氫吡喃-6-磺醯基；Pmc: 2,2.5,7,8-pentamethylchroman-6-sulfonyl;

Dde：1-(4,4-二甲基-2,6-二氧代環亞己基)乙基；Dde: 1-(4,4-dimethyl-2,6-dioxocyclohexylene)ethyl;

ivDde)：1-(4,4-二甲基-2,6-二氧代環亞己基)3-甲基丁基；ivDde): 1-(4,4-dimethyl-2,6-dioxocyclohexylene)3-methylbutyl;

Mtt：4-甲基三苯甲基；Mtt: 4-methyltrityl;

Mmt：4-甲氧基三苯甲基；Mmt: 4-methoxytrityl;

DCM：二氯甲烷；DCM: dichloromethane;

TIS：三異丙基矽烷；TIS: triisopropyl decane;

TFA：三氟乙酸；TFA: trifluoroacetic acid;

Et₂ O：***；Et ₂ O: diethyl ether;

NMP：N-甲基吡咯烷酮；NMP: N-methylpyrrolidone;

HOAt：1-羥基-7氮雜-1H-苯並***；HOAt: 1-hydroxy-7aza-1H-benzotriazole;

HOBt：1-羥基苯並***；HOBt: 1-hydroxybenzotriazole;

DIC：N,N-二異丙基碳二亞胺。DIC: N,N-diisopropylcarbodiimide.

Q的合成Synthesis of Q

用式(II)表示的Q可商業購得,為文獻已知或可採用多種本領域專業技術人員熟悉的方法方便地制得。2010年Oishi等報導了一種合成式(II)化合物的通用合成路徑(S. Oishi etc.,J. Chem. Soc.,Perkin Trans. 1,2001,2445)，其中X、Y和R₃ 均為氫(H)，如方案1所示。Q represented by formula (II) is commercially available and is conveniently prepared by the literature or by a variety of methods familiar to those skilled in the art. In 2010, Oishi et al. reported a general synthetic route for the synthesis of compounds of formula (II) (S. Oishi etc., J. Chem. Soc., Perkin Trans. 1, 2001, 2445), in which X, Y and R ₃ are Hydrogen (H), as shown in Scheme 1.

方案1plan 1

用式(II)表示的Q可商業購得，或使用文獻中已知的或採用多種本領域專業技術人員熟悉的方法方便地制得。一種合成式(II)化合物的通用合成路徑如下方案2中所示，其X是氟(F)、Y和R₃ 為氫(H)。關鍵起始原料4可商業購得，為文獻已知(T. Narumi et al.,Tetrahedron,2008,64,4332)。Q represented by formula (II) is commercially available or conveniently prepared using methods known in the literature or by a variety of methods familiar to those skilled in the art. A general synthetic route for the synthesis of compounds of formula (II) is shown in Scheme 2, wherein X is fluorine (F), Y and R ₃ are hydrogen (H). The key starting material 4 is commercially available and is known in the literature (T. Narumi et al., Tetrahedron, 2008, 64, 4332).

方案2Scenario 2

用式(IIIa)表示的Q可商業購得，為文獻中已知的或可採用多種本領域專業技術人員熟悉的方法方便地制得。一種合成式(IIIa)化合物的通用合成路徑如下方案3中所示，其X是三氟甲基(CF₃ )，Z是氮(N)，Y、R₁ 、R₂ 和R₃ 均為氫(H)。關鍵起始原料3,3,3-三氟-硝基丙烷(3,3,3-trifluorol-nitropropen)6可商業購得，為文獻已知。谷氨酸二酯對3,3,3-三氟-硝基丙烷6的Aza-邁克爾加成(Aza-Michael addition)反應是採用立體化學控制方式(M. Molteniet al., Org. Lett.,2003,5,3887)。Q represented by formula (IIIa) is commercially available and is conveniently prepared in the literature or by a variety of methods familiar to those skilled in the art. A general synthetic route for the synthesis of compounds of formula (IIIa) is shown in Scheme 3, wherein X is trifluoromethyl (CF ₃ ), Z is nitrogen (N), and Y, R ₁ , R ₂ and R ₃ are all hydrogen. (H). The key starting material, 3,3,3-trifluorol-nitropropen 6, is commercially available and is known in the literature. The Aza-Michael addition reaction of glutamic acid diester to 3,3,3-trifluoro-nitropropane 6 is controlled by stereochemistry (M. Molteni et al., Org. Lett., 2003, 5, 3887).

方案3Option 3

用式(IIIb)表示的Q可商業購得，為文獻中已知的或可採用多種本領域專業技術人員熟悉的方法方便地制得。一種合成式(IIIb)化合物的通用合成路徑如下方案4中所示，其X是三氟甲基(CF₃ )，Z是氮(N)，R₂ 為烷基，Y、R₁ 和R₃ 均為氫(H)。起始原料10可商業購得，為文獻已知(J. Andre et al.,Eur. J. Org. Chem. 2004,1558)。關鍵步驟包括三氟甲磺酸鹽(triflate)11與谷氨酸二酯7取代反應的SN₂ 立體定向問題(P. O’ Shea et a1., J. Org. Chem. 2009,5,1605)。The Q represented by the formula (IIIb) is commercially available, and is conveniently prepared by a method known in the literature or by various methods familiar to those skilled in the art. A general synthetic route for the synthesis of compounds of formula (IIIb) is shown in Scheme 4, wherein X is trifluoromethyl (CF ₃ ), Z is nitrogen (N), R ₂ is alkyl, Y, R ₁ and R ₃ All are hydrogen (H). Starting material 10 is commercially available and is known in the literature (J. Andre et al., Eur. J. Org. Chem. 2004, 1558). The key steps include the SN ₂ stereotactic problem of triflate 11 and glutamic acid diester 7 substitution reactions (P. O' Shea et al., J. Org. Chem. 2009, 5, 1605). .

方案4Option 4

此外，式(IIIb)的Q可採用另一種合成路徑方案5制得，其中X是三氟甲基(CF₃ )，Z是氮(N)，R₂ 為烷基，Y、R₁ 和R₃ 均為氫(H)。關鍵起始原料10(J. Andre,et al.,Eur. J. Org. Chem. 2004,1558)被氧化後得到三氟甲基酮(trifluoromethylketone)13。在鹼的存在條件下，可形成下述亞胺類化合物。最後一步包括亞胺14與氫硼化鈉或氫硼化鋅的立體定向還原反應，從而得到立體異構體12和15(G. Huges,et al.,Angew Chem. Int. Ed. 2007,46,1839)。Furthermore, Q of formula (IIIb) can be prepared by another synthetic route scheme 5 wherein X is trifluoromethyl (CF ₃ ), Z is nitrogen (N), R ₂ is alkyl, Y, R ₁ and R ₃ is hydrogen (H). The key starting material 10 (J. Andre, et al., Eur. J. Org. Chem. 2004, 1558) is oxidized to give trifluoromethylketone 13. The following imine compound can be formed in the presence of a base. The final step involves stereospecific reduction of imine 14 with sodium borohydride or zinc borohydride to give stereoisomers 12 and 15 (G. Huges, et al., Angew Chem. Int. Ed. 2007, 46). , 1839).

方案5Option 5

此外，式(IIIb)的Q還可以採用另一種合成路徑方案6製備，其中X是三氟甲基，Z是氮，R₂ 為烷基，Y、R₁ 和R₃ 均為氫。已知起始原料二胺16(M.Mandal,et al.,J. Am Chem. Soc. 2002,6538)與乙醛(aldehyde)17的縮聚反應可得到亞胺18。接下來，在催化量的路易斯酸存在下，亞胺18與TMSCN，發生非對映選擇性Strecker反應(diastereoselective Strecker-type reaction)。最後一步包括水解含氰基的中間體，從而獲得立體異構體12(F. Huguentt,et al.,J. Org. Chem. 2006,71,7075)。Furthermore, Q of formula (IIIb) can also be prepared by another synthetic route scheme 6, wherein X is trifluoromethyl, Z is nitrogen, R ₂ is alkyl, and Y, R ₁ and R ₃ are all hydrogen. The polycondensation reaction of the starting material diamine 16 (M. Mandal, et al., J. Am Chem. Soc. 2002, 6538) with acetaldehyde 17 is known to give the imine 18. Next, in the presence of a catalytic amount of Lewis acid, the imine 18 and TMSCN undergo a diastereoselective Strecker-type reaction. The final step involves the hydrolysis of the cyano-containing intermediate to give stereoisomer 12 (F. Huguentt, et al., J. Org. Chem. 2006, 71, 7075).

方案6Option 6

用式(IV)表示的Q可商業購得，為文獻中已知的或可採用多種本領域專業技術人員熟悉的方法方便地制得。一種合成式(IV)化合物的通用合成路徑如下方案7中所示，其X是氧(O)，Z是碳(C)，R₂ 為烷基，W、Y、R₁ 和R₃ 均為氫(H)。起始原料-酮酸酯19可商業購得，為文獻已知(R. Hoffman,et al.,J. Org. Chem. 1999,64:1558)。β-酮酸酯19和三氟甲基磺酸鹽20烷基化反應後，R₉ 去羰基和脫保護，從而得到酮亞甲基異酯21(R. Hoffman,et al.,J. Org. Chem. 1999,64:1558；P. S. Dragovich,et al.,J. Med. Chem. 1999,42:1203)。Q represented by formula (IV) is commercially available and is conveniently prepared in the literature or by a variety of methods familiar to those skilled in the art. A general synthetic route for the synthesis of compounds of formula (IV) is shown in Scheme 7, wherein X is oxygen (O), Z is carbon (C), R ₂ is alkyl, and W, Y, R ₁ and R ₃ are Hydrogen (H). The starting material - ketoester 19 is commercially available and is known in the literature (R. Hoffman, et al., J. Org. Chem. 1999, 64: 1558). After alkylation of the β-ketoester 19 with the trifluoromethanesulfonate 20, R _{9 is} decarbonylated and deprotected to give the ketomethylene isoester 21 (R. Hoffman, et al., J. Org) Chem. 1999, 64: 1558; PS Dragovich, et al., J. Med. Chem. 1999, 42: 1203).

方案7Option 7

用式(IV)表示的Q可商業購得，為文獻中已知的或可採用多種本領域專業技術人員熟悉的方法方便地制得。其中，X是氧，Z是碳，R₂ 為烷基，W是氟，Y、R₁ 和R₃ 均為氫。一種合成式(IV)化合物的通用合成路徑如下方案8中所示。起始原料三氟烷基化的-酮酸酯22可商業購得，或按照文獻(R. Hoffman,et al.,J. Org. Chem. 1999,64:1558)制得。此外，β-酮酯22與三氟甲磺酸鹽(triflate)20烷基化反應後，進行去羰基化反應，從而得到酮亞甲基異酯(ketomethylen isoester)23，然後將23轉換成相應的Z-TMS烯醇醚，與Selectfluo氟化反應，最後脫保護得到單氟代的酮亞甲基異酯25(R. Hoffman,et al.,J. Org. Chem. 1999,64:1558；P. S. Dragovich,et al.,J. Med. Chem. 1999,42:1203)。Q represented by formula (IV) is commercially available and is conveniently prepared in the literature or by a variety of methods familiar to those skilled in the art. Wherein X is oxygen, Z is carbon, R ₂ is an alkyl group, W is fluorine, and Y, R ₁ and R ₃ are all hydrogen. A general synthetic route for the synthesis of compounds of formula (IV) is shown in Scheme 8. The starting material trifluoroalkylated-ketoester 22 is commercially available or can be prepared according to the literature (R. Hoffman, et al., J. Org. Chem. 1999, 64: 1558). Further, after the alkylation reaction of the β-ketoester 22 with the triflate 20, a decarbonylation reaction is carried out to obtain a ketomethylen isoester 23, and then 23 is converted into a corresponding Z-TMS enol ether, fluorinated with Selectfluo, and finally deprotected to give monofluoro ketomethyl isoester 25 (R. Hoffman, et al., J. Org. Chem. 1999, 64: 1558; PS Dragovich, et al., J. Med. Chem. 1999, 42: 1203).

方案8Option 8

一般合成方案General synthetic scheme

連接在MBHA樹脂上的中間肽片段可採用固相多肽化學法在應用生物系統公司(Applied Biosystems Inc.,ABI)的460A型多肽合成儀上製備，該多肽合成儀使用MBHA樹脂(應用生物系統公司，lot # A1A023,0.77 mmol/g)。所有氨基酸的α-氨基用叔丁氧羰基保護。這些帶有反應側鏈的氨基酸的保護有如下幾種：精氨酸(TOS)、賴氨酸(Cl-Z)、色氨酸(CHO)、谷氨酸(CHex)、酪氨酸(溴-Z)、絲氨酸(Bzl)、天冬氨酸(OBzl)、蘇氨酸(Bzl)。The intermediate peptide fragment attached to the MBHA resin can be prepared by solid phase polypeptide chemistry on Applied Biosystems Inc. (ABI) 460A polypeptide synthesizer using MBHA resin (Applied Biosystems) , lot # A1A023, 0.77 mmol/g). The a-amino group of all amino acids is protected with a tert-butoxycarbonyl group. The protection of these amino acids with reactive side chains is as follows: arginine (TOS), lysine (Cl-Z), tryptophan (CHO), glutamic acid (CHex), tyrosine (bromine) -Z), serine (Bzl), aspartic acid (OBzl), threonine (Bzl).

被保護的氨基酸在二氯甲烷(DCM)中用0.5當量的二環己基碳二亞胺(DCC)：1當量的氨基酸激活，從而獲得該氨基酸的對稱酸酐。但是，精氨酸殘基、谷氨酸殘基和甘氨酸殘基，是通過形成這些氨基酸的1-羥基苯***(HOBt)酯進行激活(當量比1:1:1的氨基酸：HOBt：DCC的溶於二甲基甲醯胺(DMF)中)。The protected amino acid was activated in dichloromethane (DCM) with 0.5 equivalent of dicyclohexylcarbodiimide (DCC): 1 equivalent of amino acid to obtain a symmetric anhydride of the amino acid. However, arginine residues, glutamic acid residues, and glycine residues are activated by the formation of 1-hydroxybenzotriazole (HOBt) esters of these amino acids (amino acid equivalent ratio 1:1:1: HOBt: DCC Soluble in dimethylformamide (DMF)).

氨基酸殘基按順序依次從C-末端到N-末端進行連接，以一系列的脫保護週期結束。一個耦合週期包括：活化氨基酸被先前耦合的氨基酸的游離伯胺基親核取代過程。脫保護是指用無水三氟乙酸(TFA)脫除N-末端封端基團Boc，從而在二異丙基乙胺(DIEA)中和後產生一個游離氨基。The amino acid residues are sequentially linked from the C-terminus to the N-terminus, ending with a series of deprotection cycles. A coupling cycle includes a free amino acid nucleophilic substitution process of the activated amino acid by the previously coupled amino acid. Deprotection refers to the removal of the N-terminal capping group Boc with anhydrous trifluoroacetic acid (TFA) to produce a free amino group after neutralization with diisopropylethylamine (DIEA).

合成量為0.5mmol。MBHA-樹脂上的功能位點濃度為0.77mmol/g，所用樹脂量為649mg。所有氨基酸使用2倍摩爾量額外的對稱酸酐。按照標準方法，C-末端精氨酸能夠耦合在MBHA樹脂上。所有氨基酸殘基都是雙耦合的，即各個氨基酸殘基可被樹脂耦合兩次，以保證樹脂上的NH₂ 基團能夠完全反應。第二次耦合發生在在重新加入新的氨基酸前的在沒有Boc脫保護階段。這將有助於所有的樹脂游離氨基完全地反應。色氨酸殘基是四重耦合的。在各個雙耦合週期的第二次耦合之後，末端的Boc基團可用無水TFA脫除，可用DIEA中和。The amount of synthesis was 0.5 mmol. The functional site concentration on the MBHA-resin was 0.77 mmol/g, and the amount of the resin used was 649 mg. All amino acids were used in a 2-fold molar amount of additional symmetrical anhydride. The C-terminal arginine can be coupled to the MBHA resin according to standard methods. All amino acid residues are double-coupled, ie each amino acid residue can be coupled twice by a resin to ensure complete reaction of the NH ₂ group on the resin. The second coupling occurs before the new amino acid is re-added in the absence of Boc deprotection. This will help all of the free amino groups of the resin react completely. The tryptophan residue is quadruple coupled. After the second coupling of each double coupling cycle, the terminal Boc group can be removed with anhydrous TFA and neutralized with DIEA.

在從樹脂切除肽之前，可用溶於DMF中的呱啶脫除色氨酸殘基上的甲醯基側鏈封端基團。將多肽基-樹脂轉移到50mL燒結玻璃漏斗後，用DCM和DMF洗滌數次。然後向上述多肽樹脂中加入3-5mL的50/50的呱啶/DMF溶液，以剛好沒過多肽樹脂。5min後，真空除去呱啶/DMF溶液，然後再加入3-5mL的呱啶/DMF溶液。10min後，再次真空過濾除去呱啶/DMF溶液，然後加入15-20mL的呱啶/DMF溶液，在15min後，除去呱啶/DMF，所得多肽基-樹脂依次用DMF和DCM洗滌數次。最後，所得多肽-樹脂然後用置於真空乾燥箱(不加熱)中，完全乾燥除去溶劑。The formazan side chain capping group on the tryptophan residue can be removed with acridine dissolved in DMF prior to excision of the peptide from the resin. The polypeptide-resin was transferred to a 50 mL sintered glass funnel and washed several times with DCM and DMF. Then, 3-5 mL of a 50/50 acridine/DMF solution was added to the above polypeptide resin so that the polypeptide resin was not passed. After 5 min, the acridine/DMF solution was removed in vacuo and then 3-5 mL of acridine/DMF solution was added. After 10 min, the acridine/DMF solution was again removed by vacuum filtration, then 15-20 mL of acridine/DMF solution was added. After 15 min, acridine/DMF was removed and the resulting polypeptide-resin was washed several times with DMF and DCM. Finally, the resulting polypeptide-resin was then placed in a vacuum oven (without heating) and completely dried to remove the solvent.

此外，所需要的聚合物連接的肽鍵片段可以通過Fmoc保護制得。Rink Amide MBHA樹脂，Fmoc保護氨基酸，O-苯並***-1-基-N,N,N’,N’-四甲基-脲六氟磷酸酯(HBTU))溶於N,N-二甲基甲醯胺(DMF)，N-甲基嗎啉(NMM)活化，呱啶脫Fmoc保護(步驟1)。必要時，可以人工地對賴氨酸Lys(Aloc)基團進行選擇性脫保護，將樹脂用3倍當量的鈀(三苯基膦)(Pd(PPh₃ )₄ )溶於5mL的CHCl₃ ：NMM：HOAc(18：1：0.5)溶液處理2小時，從而完成對賴氨酸基團的選擇性脫保護。然後，所得樹脂殘留物用CHC1₃ (6X5 mL)洗滌，20%的HOAc的DCM(6X5 mL)的溶液洗滌，DCM(6X5 mL)洗滌，最後用DMF(6X5 mL)洗滌。在一些例子中，在此之後，該化合物的合成因為2-(2-(2-氨基乙氧基)乙氧基)乙酸(AEEA)基團、乙酸或3-馬來醯亞胺基丙酸(MPA)的實現自動重複加入(步驟3)。用85%TFA/5%TIS/5%茴香硫醚和5%苯酚體系完成樹脂切割和產品分離，然後用乾冰冷凍的***(Et₂ O)析出產品(步驟4)。所得產品用適用Varian(Rainin)二元HPLC系統的製備型HPLC分離純化：30-55%的B相[0.045% TFA的H₂ O溶液(A相)；0.045%TFA的CH₃ CN溶液(B相)]梯度洗脫、流速9.5mL/min、洗脫時間180 min、使用Phenomenex Luna 10μ phenyl-hexyl(21mm x 25cm)柱、UV檢測器(Varian Dynamax UVD II)在波長214 nm和254nm處檢測。用RP-HPLC質譜定量分析產品純度為95%，該RP-HPLC質譜使用惠普LCMS-1100-二極管陣列檢測器分光儀(Hewlett Packard LCMS-1100 series spectrometer with a diode array detector)和電噴霧離子阱。In addition, the desired polymer-linked peptide bond fragments can be prepared by Fmoc protection. Rink Amide MBHA resin, Fmoc protected amino acid, O-benzotriazol-1-yl-N,N,N',N'-tetramethyl-urea hexafluorophosphate (HBTU)) soluble in N,N-II Methylformamide (DMF), N-methylmorpholine (NMM) is activated, and acridine is deprotected by Fmoc (step 1). If necessary, the lysine Lys (Aloc) group can be selectively deprotected artificially, and the resin is dissolved in 5 mL of CHCl ₃ with 3 equivalents of palladium (triphenylphosphine) (Pd(PPh ₃ ) ₄ ). :NMM: HOAc (18:1:0.5) solution treatment for 2 hours to complete selective deprotection of the lysine group. Then, the resulting resin was washed residue CHC1 ₃ (6X5 mL), washed 20% HOAc solution of DCM (6X5 mL) of, DCM (6X5 mL) was used, and finally washed with DMF (6X5 mL). In some instances, after this synthesis, the compound is synthesized because of the 2-(2-(2-aminoethoxy)ethoxy)acetic acid (AEEA) group, acetic acid or 3-maleimidopropionic acid. The implementation of (MPA) is automatically repeated (step 3). Resin cleavage and product separation were carried out with 85% TFA / 5% TIS / 5% thioanisole and 5% phenol system, and then the product was precipitated from dry ice chilled diethyl ether (Et ₂ O) (step 4). The resulting product isolated and purified by preparative HPLC applicable Varian (Rainin) binary HPLC system: 30 to 55% of Phase B [0.045% TFA in H ₂ O solution (A-phase); 0.045% TFA solution in CH ₃ CN (B Phase)] Gradient elution, flow rate 9.5 mL/min, elution time 180 min, detection using Phenomenex Luna 10μ phenyl-hexyl (21mm x 25cm) column, UV detector (Varian Dynamax UVD II) at 214 nm and 254 nm . The purity of the product was determined by RP-HPLC mass spectrometry to be 95%. The RP-HPLC mass spectrometer used a Hewlett Packard LCMS-1100 series spectrometer with a diode array detector and an electrospray ion trap.

保護基是指那些用於保護多肽衍生物從而阻止其自身發生反應的化學片段。這些保護基包括：乙醯基，9-芴甲氧羰基(FMOC)、叔丁氧羰基(Boc)，苄氧羰基(CBZ)和類似的保護劑。氨基酸的專用保護基見表1。Protecting groups are those chemical fragments that are used to protect a polypeptide derivative from its own reaction. These protecting groups include: ethyl fluorenyl, 9-fluorenylmethoxycarbonyl (FMOC), tert-butoxycarbonyl (Boc), benzyloxycarbonyl (CBZ) and similar protective agents. The specific protection groups for amino acids are shown in Table 1.

Q-連接子-a：(2S,5R)-2-(3-(叔丁氧羰基氨基)-丁烯-1-基)戊二酸-5-叔丁酯的製備Preparation of Q-linker-a: (2S,5R)-2-(3-(tert-Butoxycarbonylamino)-buten-1-yl)glutaric acid-5-tert-butyl ester

參考文獻： J. Chem. Soc.,Perkin Trans. 1,2001,2445 References: J. Chem. Soc., Perkin Trans. 1, 2001, 2445

將370mg(1mmol)的(2S,5R)-2-((3-叔丁氧羰基氨基)丁烯-1-基)戊二酸-5-叔丁酯-1-甲酯溶於2 mL甲醇中，在室溫下加入2mL(1M)氫氧化鋰(LiOH)溶液處理1小時。真空旋蒸除去大部分溶劑後加水(10mL)稀釋，調pH至5，水相用乙酸乙酯(3X30ml)萃取，得泡狀產品320mg，收率90%。370 mg (1 mmol) of (2S,5R)-2-((3-tert-butoxycarbonylamino)buten-1-yl)glutaric acid-5-tert-butyl ester-1-methyl ester was dissolved in 2 mL of methanol The solution was added to a solution of 2 mL (1 M) of lithium hydroxide (LiOH) at room temperature for 1 hour. Most of the solvent was removed by vacuum distillation, and then diluted with water (10 mL) to adjust pH to 5, and the aqueous phase was extracted with ethyl acetate (3×30 ml) to afford 320 mg of the product.

表徵資料如下：¹ H NMR：5.43(m,1H),5.33(dd,J=15.5,5.2 Hz,1H),4.59(d,J=7.6Hz,1H),3.88(m,1H),2.91(m,1H),2.25(m,2H),1.91-2.04(m,1H),1.67-1.80(m,1H),1.57(s,9H),1.47(s,9H),1.14(d,J=6.7 Hz,3H)；LCMS：358(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 5.43 (m, 1 H), 5.33 (dd, J = 15.5, 5.2 Hz, 1H), 4.59 (d, J = 7.6 Hz, 1H), 3.88 (m, 1H), 2.91 ( m, 1H), 2.25 (m, 2H), 1.91-2.04 (m, 1H), 1.67-1.80 (m, 1H), 1.57 (s, 9H), 1.47 (s, 9H), 1.14 (d, J = 6.7 Hz, 3H); LCMS: 358 (M+H) ⁺ .

Q-連接子-b：(2S,5R)-2-(3-(叔丁氧羰基氨基)-2-氟丁烯-1-基)戊二酸-5-叔丁酯的製備Preparation of Q-linker-b: (2S,5R)-2-(3-(tert-Butoxycarbonylamino)-2-fluorobuten-1-yl)glutaric acid-5-tert-butyl ester

步驟A：2S,5R-2-(3-叔丁氧基羰基氨基-2-氟-丁烯-1-基)-戊二酸1-(S)磺內醯胺的製備Step A: Preparation of 2S,5R-2-(3-tert-butoxycarbonylamino-2-fluoro-buten-1-yl)-glutaric acid 1-(S)sulfonamide

參考文獻： Tetrahedron,2008,64:4332。 References: Tetrahedron, 2008, 64: 4332.

將502mg(1mmol)的5-叔丁氧基羰基氨基-4-氟-2-(3-羥基-丙基)-己烯-3-酸(S)磺內醯胺溶於5mL的DMF中，加入2.5m_m ol的重鉻酸吡啶(PDC)，反應液常溫攪拌64小時。然後，反應混合物用20ml鹽水(brine)稀釋，用乙酸乙酯(3X20ml)萃取。合併有機相，用MgSO₄ 乾燥，減壓蒸乾溶劑。所得殘留物用快速柱色譜純化，得到425mg泡狀中間產物酸(79%)，此產品直接用於下一步反應。502 mg (1 mmol) of 5-tert-butoxycarbonylamino-4-fluoro-2-(3-hydroxy-propyl)-hexene-3-acid (S) sulphonamide was dissolved in 5 mL of DMF. 2.5 _m mol of dichromate pyridine (PDC) was added, and the reaction solution was stirred at room temperature for 64 hours. The reaction mixture was then diluted with EtOAc (EtOAc)EtOAc. The combined organic layers were dried with MgSO ₄ and evaporated The residue obtained was purified by flash column chromatography to afford </RTI></RTI><RTIgt;

步驟B：2S,5R-2-(3-叔丁氧基羰基氨基-2-氟-丁烯-1-基)-戊二酸5-tert-丁基酯1-(S)磺內醯胺的製備Step B: 2S,5R-2-(3-tert-Butoxycarbonylamino-2-fluoro-buten-1-yl)-glutaric acid 5-tert-butyl ester 1-(S) sulphonamide Preparation

從步驟A得到的400mg(0.75mmol)的2S,5R-2-(3-叔丁氧基羰基氨基-2-氟-丁烯-1-基)-戊二酸2-(S)磺內醯胺溶於10mL的二氯甲烷中，分別加入叔丁醇(0.5ml，10當量)、DCC(1.5mmol)和DMAP(1.5mmol)處理。反應混合物常溫攪拌24小時，然後，用20mL鹽水(brine)稀釋，乙酸乙酯(3X20ml)萃取。合併有機相，用MgSO₄ 乾燥，所得溶液減壓蒸乾。所得殘留物用快速柱色譜純化得到425mg泡狀叔丁酯(79%)。400 mg (0.75 mmol) of 2S,5R-2-(3-tert-butoxycarbonylamino-2-fluoro-buten-1-yl)-glutaric acid 2-(S)sulfonate obtained from Step A The amine was dissolved in 10 mL of dichloromethane and treated with t-butanol (0.5 mL, 10 eq.), DCC (l. The reaction mixture was stirred at room temperature for 24 hours, then diluted with 20 mL of brine and ethyl acetate (3×20 mL). The organic layers were combined, dried over MgSO ₄ and evaporated The residue obtained was purified by flash column chromatography toield </

表徵資料如下：¹ H NMR：5.33(m,1H)、4.54(m,1H)、3.88(m,1H)、3.37(s,2H)、3.23(m,1H)、2.25(m,2H)、1.91-2.14(m,4H)、1.67-1.80(m,5H)、1.57(s,9H)、1.47(d,J=7.6Hz,3H)、1.18(s,3H)、1.14(s,3H)；LCMS：574(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 5.33 (m, 1H), 4.54 (m, 1H), 3.88 (m, 1H), 3.37 (s, 2H), 3.23 (m, 1H), 2.25 (m, 2H), 1.91-2.14 (m, 4H), 1.67-1.80 (m, 5H), 1.57 (s, 9H), 1.47 (d, J = 7.6 Hz, 3H), 1.18 (s, 3H), 1.14 (s, 3H) LCMS: 574 (M+H) ⁺ .

步驟C：2S,5R-2-(3-叔丁氧基羰基氨基-2-氟-丁烯-1-基)-戊二酸5-叔丁酯的製備Step C: Preparation of 2S,5R-2-(3-tert-butoxycarbonylamino-2-fluoro-buten-1-yl)-glutaric acid 5-tert-butyl ester

將步驟B得到的叔丁酯(410mg,0.72mmol)和50%H₂ O₂ 水溶液(260ml,3.6mmol)溶於THF-H₂ O(5:1,12mL)中，在溫度0℃下加入LiOH(1N,1.44mL)。所得混合物室溫攪拌2小時。然後，所得混合物調pH至5，用乙酸乙酯(3X15mL)萃取，合併有機相用鹽水洗滌，用MgSO₄ 乾燥。所得溶液減壓蒸乾得到相應的發泡狀酸(262mg,95%)。The tert-butyl ester obtained in Step B (410 mg, 0.72 mmol) and 50% aqueous H ₂ O ₂ (260 mL, 3.6 mmol) was dissolved in THF-H ₂ O (5:1, 12 mL) LiOH (1 N, 1.44 mL). The resulting mixture was stirred at room temperature for 2 hours. Then, the resulting mixture was adjusted to pH 5, and extracted with ethyl acetate (3 X 15 mL), the combined organic phases were washed with brine, dried over MgSO _4. The resulting solution was evaporated to dryness to dryness crystals crystals crystals

表徵資料如下：¹ H NMR：5.23(m,1H)、4.45(m,1H)、3.11(m,1H)、2.45(m,2H)、2.24(m,2H)、1.52(s,9H)、1.47(s,9H)；LCMS：376(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 5.23 (m, 1H), 4.45 (m, 1H), 3.11 (m, 1H), 2.45 (m, 2H), 2.24 (m, 2H), 1.52 (s, 9H), 1.47 (s, 9H); LCMS: 376 (M+H) ⁺ .

Q-連接子-c：2R,5R 2-[1-(叔丁氧基羰基氨基-甲基)-2,2,2-三氟-乙基氨基]-戊二酸5-叔丁酯的製備Q-linker-c: 2R, 5R 2-[1-(tert-butoxycarbonylamino-methyl)-2,2,2-trifluoro-ethylamino]-glutaric acid 5-tert-butyl ester preparation

步驟A：2R,5R 2-[1-(叔丁氧基羰基氨基-甲基)-2,2,2-三氟-乙基氨基]-戊二酸5-叔丁酯1-甲酯的製備Step A: 2R,5R 2-[1-(tert-Butoxycarbonylamino-methyl)-2,2,2-trifluoro-ethylamino]-glutaric acid 5-tert-butyl ester 1-methyl ester preparation

參考文獻：Org. Lett.,2003,5,3887。References: Org. Lett., 2003, 5, 3887.

將364mg(1mol)的2-(1-氨甲基-2,2,2-三氟-乙基氨基)-戊二酸5-叔丁酯1-甲酯鹽酸鹽和260mg(1.2mmol)的Boc₂ O溶於15mL的DCM中，在溫度0℃下加入DIPEA(0.2mL,1.5mmol)的DCM(1mL)溶液。反應液常溫攪拌6小時。所得混合物用30mL的乙酸乙酯稀釋後，分別用0.1N鹽酸和鹽水(brine)洗滌，用MgSO₄ 乾燥有機相。所得溶液減壓蒸乾後，用快速柱色譜法純化，獲得相應的發泡狀二酯(420mg，85%)。364 mg (1 mol) of 2-(1-aminomethyl-2,2,2-trifluoro-ethylamino)-glutaric acid 5-tert-butyl ester 1-methyl ester hydrochloride and 260 mg (1.2 mmol) the Boc DCM ₂ O was dissolved in 15mL was added at a temperature of 0 ℃ DIPEA (0.2mL, 1.5mmol) in DCM (1mL) was added. The reaction solution was stirred at room temperature for 6 hours. The resulting mixture was diluted with 30mL ethyl acetate, was washed (brine) with 0.1N hydrochloric acid and brine, the organic phase was dried over MgSO _4. The resulting solution was evaporated to dryness crystals crystals crystals crystals

表徵資料如下：¹ H NM：4.54(m,1H)、4.12(m,1H)、3.68(s,3H)、3.45(m,1H)、3.11(m,2H)、2.45(m,2H)、2.24(m,2H)、1.52(s,9H)、1.47(s,9H)；LCMS：430(M+H)⁺ 。The characterization data are as follows: ¹ H NM: 4.54 (m, 1H), 4.12 (m, 1H), 3.68 (s, 3H), 3.45 (m, 1H), 3.11 (m, 2H), 2.45 (m, 2H), 2.24 (m, 2H), 1.52 (s, 9H), 1.47 (s, 9H); LCMS: 430 (M+H) ⁺ .

步驟B：2R,5RStep B: 2R, 5R 2-[1-(叔丁氧基羰基氨基-甲基)-2,2,2-三氟-乙基氨基]-戊二酸5-叔丁酯的製備Preparation of 2-[1-(tert-Butoxycarbonylamino-methyl)-2,2,2-trifluoro-ethylamino]-glutaric acid 5-tert-butyl ester

將步驟A得到的叔丁酯(420mg,0.92mmol)溶於THF-H₂ O(5:1,12mL)中,在溫度0℃下加入LiOH(1N,1.44mL)。反應液室溫攪拌2小時。所得混合物調pH至5，用乙酸乙酯(3X15mL)萃取。合併有機相，用鹽水(brine)洗滌，MgSO₄ 乾燥。所得溶液減壓蒸乾得到相應的酸(362mg,92%)。The tert-butyl ester obtained in Step A (420 mg, 0.92 mmol) was dissolved in THF-H ₂ O (5:1, 12 mL). The reaction solution was stirred at room temperature for 2 hours. The mixture was adjusted to pH 5 and extracted with ethyl acetate (3×15 mL). The combined organic phases were dried washed (brine) saline over MgSO _4. The resulting solution was evaporated to dryness crystals crystals crystals

表徵資料如下：¹ H NMR：4.50(m,1H)、4.08(m,1H)、3.45(m,1H)、3.11(m,2H)、2.45(m,2H)、2.24(m,2H)、1.52(s,9H)、1.47(s,9H)；LCMS：430(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 4.50 (m, 1H), 4.08 (m, 1H), 3.45 (m, 1H), 3.11 (m, 2H), 2.45 (m, 2H), 2.24 (m, 2H), 1.52 (s, 9H), 1.47 (s, 9H); LCMS: 430 (M+H) ⁺ .

Q-連接子-d：2R-2-(1S,2S-2-叔丁氧基羰基氨基-1-三氟-甲基-丙基氨基)-戊二酸5-丁基酯的製備Preparation of Q-linker-d:2R-2-(1S,2S-2-tert-butoxycarbonylamino-1-trifluoro-methyl-propylamino)-pentanedioic acid 5-butyl ester

步驟A：2S,3S-3-二苯基氨基-1,1,1-三氟丁烷-2-三氟甲烷磺酸鹽的製備的製備Step A: Preparation of 2S,3S-3-diphenylamino-1,1,1-trifluorobutane-2-trifluoromethanesulfonate

參考文獻：Eur. J. Org. Chem. 2004,1558。References: Eur. J. Org. Chem. 2004, 1558.

將3.23g(10mmol)的2S,3S-3-二苯基氨基-1,1,1-三氟-丁-2-醇和1.7g(16mmol)的2,6-二甲基吡啶溶於25ml環己烷中，在溫度10℃下加入4.2g(15mmol)三氟甲磺酸酐，並控制加入速度以保持溫度在10℃以下，持續反應1.5小時。所得反應混合物用水(25ml)和環己烷(50ml)稀釋。有機相分別用1N鹽酸(2X15ml)和鹽水(15ml)洗滌，用MgSO₄ 乾燥，所得溶液減壓蒸乾得到相應的三氟甲磺酸酯(4.34g,96%)。3.23 g (10 mmol) of 2S,3S-3-diphenylamino-1,1,1-trifluoro-butan-2-ol and 1.7 g (16 mmol) of 2,6-lutidine were dissolved in a 25 ml ring In hexane, 4.2 g (15 mmol) of trifluoromethanesulfonic anhydride was added at a temperature of 10 ° C, and the addition rate was controlled to keep the temperature below 10 ° C, and the reaction was continued for 1.5 hours. The resulting reaction mixture was diluted with water (25 mL) and hexane (50ml). The organic phase was washed with 1N hydrochloric acid (2 x 15 mL) and brine (15ml), dried with MgSO _4, the resulting solution was evaporated to dryness under reduced pressure to give the corresponding triflate (4.34g, 96%).

步驟B：2S-2-(1S,2S-2-二苯基氨基-1-三氟甲基-p丙基氨基)-戊二酸1-苯甲酯5-叔丁酯的製備Step B: Preparation of 2S-2-(1S,2S-2-diphenylamino-1-trifluoromethyl-ppropylamino)-glutaric acid 1-phenylmethyl 5-tert-butyl ester

將碳酸鉀(2.08g,15mmol)加入到步驟A得到的三氟甲磺酸鹽(4.55g,10mmol)的環正己烷(25ml)溶液中。所得混合物加熱到65~70℃並保持24小時。冷卻至室溫並加水(25ml)和環正己烷(50ml)稀釋，攪拌10min，分層，所得有機相分別用1N HCl(2X15ml)和鹽水(15ml)洗滌，MgSO₄ 乾燥後，所得溶液減壓濃縮得到相應的酯(5.88g,95%)。Potassium carbonate (2.08 g, 15 mmol) was added to a solution of the trifluoromethanesulfonate (4.55 g, 10 mmol The resulting mixture was heated to 65-70 ° C for 24 hours. After cooling to room temperature and add water (25ml) and hexane ring (50ml) was diluted and stirred for 10min, layers were separated and the organic phase was washed with 1N HCl (2X15ml) and brine (15ml), dried MgSO _4, the resultant solution under reduced pressure Concentration gave the corresponding ester (5.88 g, 95%).

步驟C：2R-2-(1S,2S-2-叔丁氧基羰基氨基-1-三氟甲基丙基氨基)-戊二酸5-叔丁酯的製備Step C: Preparation of 2R-2-(1S,2S-2-tert-butoxycarbonylamino-1-trifluoromethylpropylamino)-glutaric acid 5-tert-butyl ester

取5.4g(9mmol)的步驟B得到的2S-2-(1S,2S-2-二苯基氨基-1-三氟甲基-丙基氨基)-戊二酸1-苯甲酯5-叔丁基酯，在50mL甲醇和0.9g的Pd/C存在、壓力50psi下氫化反應24小時。過濾除去催化劑，濾液真空濃縮。所得殘留物溶解於50mL的二氯甲烷中，在溫度0℃下用Boc₂ O(2.60g,12mmol)的二氯甲烷(25ml)溶液處理，隨後加入DIPEA(2mL,15mmol)的二氯甲烷(10mL)溶液，混合物室溫攪拌6小時。所得混合物用乙酸乙酯(60ml)稀釋，並分別用1N HCl，鹽水水洗滌，MgSO₄ 乾燥，所得溶液減壓濃縮後用快速色譜法分離得到發泡狀目標化合物。5.4 g (9 mmol) of Step 2 B obtained 2S-2-(1S,2S-2-diphenylamino-1-trifluoromethyl-propylamino)-pentanedioic acid 1-phenylmethyl ester 5-un The butyl ester was hydrogenated in the presence of 50 mL of methanol and 0.9 g of Pd/C at a pressure of 50 psi for 24 hours. The catalyst was removed by filtration and the filtrate was concentrated in vacuo. The residue was dissolved in 50mL of dichloromethane, treated with a solution (25ml) with Boc ₂ O (2.60g, 12mmol) in dichloromethane at 0 deg.] C, followed by addition of DIPEA (2mL, 15mmol) in dichloromethane ( 10 mL) solution, the mixture was stirred at room temperature for 6 hours. The resulting mixture was diluted with ethyl acetate (60ml), and each with 1N HCl, water, brine, dried MgSO _4, the resulting solution was concentrated under reduced pressure to give the title compound with a foamed isolated by flash chromatography.

Q-連接子-e：2S-2-(3S-3-叔丁氧基羰基氨基-2-氧-丁基)-戊二酸5-叔丁酯的製備Preparation of Q-linker-e: 2S-2-(3S-3-tert-butoxycarbonylamino-2-oxo-butyl)-glutaric acid 5-tert-butyl ester

參考文獻：J. Med. Chem. 1999,42:1203；Bioorg. Med. Chem. 2005,13:5240References: J. Med. Chem. 1999, 42: 1203; Bioorg. Med. Chem. 2005, 13: 5240

將387mg(1mmol)的2S-2-(3S-3-叔丁氧基羰基氨基-2-氧-丁基)-戊二酸5-叔丁酯1-甲酯溶於THF-H₂ O(5：1,12ml)溶液中，在溫度0℃下加入氫氧化鋰(1N,1.44ml)，混合物室溫攪拌2小時。然後，混合物調節pH至5，用乙酸乙酯(3X15ml)萃取，合併有機相，用鹽水洗滌，MgSO₄ 鎂乾燥，所得溶液減壓蒸幹得到發泡狀目標化合物酸(362mg,92%)。387 mg (1 mmol) of 2S-2-(3S-3-t-butoxycarbonylamino-2-oxo-butyl)-glutaric acid 5-tert-butyl ester 1-methyl ester was dissolved in THF-H ₂ O ( To a solution of 5:1, 12 ml), lithium hydroxide (1N, 1.44 ml) was added at a temperature of 0 ° C, and the mixture was stirred at room temperature for 2 hours. Then, the mixture was adjusted to pH 5, and extracted with ethyl acetate (3 X 15ml), the combined organic phases were washed with brine, MgSO ₄ magnesium sulfate, and the resulting solution was evaporated to dryness under reduced pressure to give the title compound foamed acid (362mg, 92%).

表徵資料如下：¹ H NMR：4.63(m,1H)、4.38(br,1H)、2.68(d,J=7.6 Hz,2H)、2.58(m,1H)、2.25(dd,J=12.3,7.6 Hz,2H)、1.92(m,2H)、1.49(s,9H)、1.47(s,9H)、1.41(d,J=7.6 Hz,3H)；LCMS：374(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 4.63 (m, 1 H), 4.38 (br, 1H), 2.68 (d, J = 7.6 Hz, 2H), 2.58 (m, 1H), 2.25 (dd, J = 12.3, 7.6 Hz, 2H), 1.92 (m, 2H), 1.49 (s, 9H), 1.47 (s, 9H), 1.41 (d, J = 7.6 Hz, 3H); LCMS: 374 (M+H) ⁺ .

Q-連接子-f：2R-2-(1S,3S-3-叔丁氧基羰基氨基-1-氟-2-氧-丁基)-戊二酸5-叔丁酯的製備Preparation of Q-linker-f: 2R-2-(1S,3S-3-tert-butoxycarbonylamino-1-fluoro-2-oxo-butyl)-glutaric acid 5-tert-butyl ester

步驟A：2S-2-(1S,3S-3-叔丁氧基羰基氨基-1-氟-2-氧-丁基)-戊二酸5-叔丁酯1-甲酯的製備Step A: Preparation of 2S-2-(1S,3S-3-tert-butoxycarbonylamino-1-fluoro-2-oxo-butyl)-glutaric acid 5-tert-butyl ester 1-methyl ester

parr振盪器裝置中，壓力50psi條件下，將581mg(1mmol)的2S-2-[1S,3S-氟-2-氧-3-(三苯甲基-氨基)-丁基]-戊二酸5-苯甲酯1-甲酯和10%Pd/C(100mg)在甲醇(25ml)中，進行氫化反應6小時。用矽藻土過濾除去催化劑，濾液濃縮，殘留物溶於二氧六環(25ml)並用1N NaOH溶液(1.2ml)處理。在溫度0℃下，將Boc₂ O(238mg,1.1mmol)的二氯甲烷(2ml)溶液加入到上述溶液中，混合物室溫攪拌6小時。多的混合物用乙酸乙酯(30ml)稀釋，分別用0.1NHCl和鹽水洗滌，並用無水硫酸鈉乾燥。所得溶液減壓蒸發，殘留物溶於二氯甲烷(10ml)並加叔丁醇(0.5ml,10當量)、DCC(1.5mmol)和DMAP(1.5mmol)處理。所得反應混合物攪拌24小時後，用鹽水(20ml)稀釋，乙酸乙酯(3X20ml)萃取。合併有機相並用MgSO₄ 乾燥，溶液減壓蒸發，殘留物用快速柱色譜法純化得發泡狀叔丁酯(315mg,66%)。In a parr shaker unit, 581 mg (1 mmol) of 2S-2-[1S,3S-fluoro-2-oxo-3-(trityl-amino)-butyl]-glutaric acid at 50 psi 5-Benzylmethyl ester 1-methyl ester and 10% Pd/C (100 mg) were subjected to hydrogenation in methanol (25 ml) for 6 hours. The catalyst was removed by filtration over EtOAc (EtOAc)EtOAc. A solution of Boc ₂ O (238 mg, 1.1 mmol) in dichloromethane (2 ml) was added to the above mixture, and the mixture was stirred at room temperature for 6 hours. The mixture was diluted with EtOAc (30 mL). The resulting solution was evaporated <RTI ID=0.0></RTI> EtOAc m. The reaction mixture was stirred for 24 h then diluted with EtOAc EtOAc m. The organic phases were combined and dried over MgSO _4, the solution was evaporated under reduced pressure, the residue was purified by flash column chromatography to give tert-butyl-shaped foam (315mg, 66%).

表徵資料如下：¹ H NMR：4.81(m,1H)、4.63(m,1H)、4.38(br,1H)、3.67(s,3H)、2.78(m,1H)、2.35(dd,J=12.3,7.6 Hz,2H)、2.06(m,2H)、1.49(s,9H)、1.47(s,9H)、1.41(d,J=7.6 Hz,3H)；LCMS：406(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 4.81 (m, 1 H), 4.63 (m, 1 H), 4.38 (br, 1H), 3.67 (s, 3H), 2.78 (m, 1H), 2.35 (dd, J = 12.3) , 7.6 Hz, 2H), 2.06 (m, 2H), 1.49 (s, 9H), 1.47 (s, 9H), 1.41 (d, J = 7.6 Hz, 3H); LCMS: 406 (M+H) ⁺ .

步驟B：2S-2-(1S,3S-3-叔丁氧基羰基氨基-1-氟-2-氧-丁基)-戊二酸5-叔丁酯的製備Step B: Preparation of 2S-2-(1S,3S-3-tert-butoxycarbonylamino-1-fluoro-2-oxo-butyl)-glutaric acid 5-tert-butyl ester

將260mg(0.65mmol)的步驟A得到的2S-2-(1S,3S-3-叔丁氧基羰基氨基-1-氟-2-氧-丁基)-戊二酸5-叔丁酯1-甲酯溶於THF-H² O(5：1,12mL)溶液中,在溫度0℃下加入LiOH(1N,1.0ml)，所得混合物室溫攪拌2小時。然後調pH至5，用乙酸乙酯(3X15ml)萃取，合併有機相，用鹽水洗滌，用MgSO₄ 乾燥。所得溶液減壓蒸乾得到泡狀目標化合物酸(238mg,95%)。260 mg (0.65 mmol) of 2S-2-(1S,3S-3-tert-butoxycarbonylamino-1-fluoro-2-oxo-butyl)-glutaric acid 5-tert-butyl ester 1 obtained in Step A The methyl ester was dissolved in THF-H ² O (5:1, 12 mL). Then adjusted to pH 5, and extracted with ethyl acetate (3 X 15ml), the combined organic phases were washed with brine, dried over MgSO _4. The resulting solution was evaporated to dryness to dryness crystall

表徵資料如下：¹ H NMR：4.81(m,1H)、4.63(m,1H)、4.38(br,1H)、2.78(m,1H)、2.35(dd,J=12.3,7.6 Hz,2H)、2.06(m,2H)、1.49(s,9H)、1.47(s,9H)、1.41(d,J=7.6 Hz,3H)；LCMS：392(M+H)⁺ 。The characterization data are as follows: ¹ H NMR: 4.81 (m, 1H), 4.63 (m, 1H), 4.38 (br, 1H), 2.78 (m, 1H), 2.35 (dd, J = 12.3, 7.6 Hz, 2H), 2.06 (m, 2H), 1.49 (s, 9H), 1.47 (s, 9H), 1.41 (d, J = 7.6 Hz, 3H); LCMS: 392 (M+H) ⁺ .

實施例1Example 1

合成synthesis

[Q-連接子-d8,谷氨酸22]GLP-1-(7-37)-肽[Q-linker-d8, glutamic acid 22] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

100μm的上述肽類似物的固相多肽合成是採用人工固相多肽合成法和使用Symphony多肽合成儀製備。其所用的Symphony多肽合成儀是使用：Fmoc保護的Rink Amide MBHA樹脂；Fmoc保護的氨基酸；溶解在N,N-二甲基甲醯胺中(DMF)中並被N-甲基嗎啡啉(NMM)活化的O-苯並三氮唑-1-基-N,N,N’,N’-四甲基脲六氟磷酸酯(HBTU)；以及用六氫吡啶脫除Fmoc保護基(步驟1)。在步驟2的產品中，Boc基在與Fmoc-His(Trt)-OH偶聯前發生裂解，使用85%TFA/5%Tis/5%苯甲硫醚/5%苯酚完成樹脂裂解和產物分離，然後用乾冰冰凍***沉澱(步驟2)。採用Varian(rainnin)製備型二元HPLC色譜體系的反相製備色譜純化產品：用30-55%的B相(含有0.045% TFA的乙腈溶液)和A相(含0.045%TFA的水溶液)進行梯度洗脫，流速9.5 mL/min，梯度洗脫時間180 min，Phenomenex Luna 10μm phenyl-hexy色譜柱、21mm×25cm、UV檢測器(Varian Dynamax UVD II)、檢測波長214nm和254nm，獲得目標肽。該目標肽的純度>95%，使用RP-HPLC確定其純度。Solid phase polypeptide synthesis of 100 [mu]m of the above peptide analogs was prepared using artificial solid phase polypeptide synthesis and using a Symphony polypeptide synthesizer. The Symphony peptide synthesizer used was: Fmoc-protected Rink Amide MBHA resin; Fmoc-protected amino acid; dissolved in N,N-dimethylformamide (DMF) and N-methylmorpholine (NMM) Activated O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU); and removal of the Fmoc protecting group with hexahydropyridine (Step 1 ). In the product of step 2, the Boc group is cleaved prior to coupling with Fmoc-His(Trt)-OH, and resin cracking and product separation are accomplished using 85% TFA/5% Tis/5% thioanisole/5% phenol. It is then precipitated with dry ice in ice (step 2). Purification of the product by reverse phase preparative chromatography using a Varian (rainnin) preparative binary HPLC chromatography system: gradient with 30-55% phase B (acetic acid solution containing 0.045% TFA) and phase A (aqueous solution containing 0.045% TFA) Elution, flow rate 9.5 mL/min, gradient elution time 180 min, Phenomenex Luna 10 μm phenyl-hexy column, 21 mm × 25 cm, UV detector (Varian Dynamax UVD II), detection wavelength 214 nm and 254 nm, the target peptide was obtained. The purity of the target peptide was >95%, and its purity was determined using RP-HPLC.

MS實測值：3412,MS理論值：3412.MS measured value: 3412, MS theoretical value: 3412.

實施例2Example 2

合成synthesis

[Q-連接子-a8-9,谷氨酸22]GLP-1-(7-37)-肽[Q-linker-a8-9, glutamic acid 22] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

合成目標GLP-1類似物使用與實施例1描述的相同序列和相同條件。The same sequence and the same conditions as described in Example 1 were used to synthesize the target GLP-1 analog.

MS實測值：1113(M+3H)³⁺ ，MS理論值：1113(M+3H)³⁺ 。MS found: 1113 (M+3H) ³⁺ , MS ^mp .

實施例3Example 3

合成synthesis

[Q-連接子-b8-9，谷氨酸22]GLP-1-(7-37)-肽[Q-linker-b8-9, glutamic acid 22] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

MS實測值：1119(M+3H)³⁺ ，MS理論值：1119(M+3H)³⁺ 。MS found: 1119 (M+3H) ³⁺ , MS: 1 1 (M+3H) ³⁺ .

實施例4Example 4

合成synthesis

[Q-連接子-c8，谷氨酸22]GLP-1-(7-37)-肽[Q-linker-c8, glutamic acid 22] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

MS實測值：3398,MS理論值：3398。MS found: 3398, MS: 3398.

實施例5Example 5

合成synthesis

[Q-連接子-e8-9,谷氨酸22]GLP-1-(7-37)-肽[Q-linker-e8-9, glutamic acid 22] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

MS實測值：1118(M+3H)³⁺ ，MS理論值：1118(M+3H)³⁺ 。MS found: 1118 (M+3H) ³⁺ , MS: 1118 (M+3H) ³⁺ .

實施例6Example 6

合成synthesis

[Q-連接子-f8-9,精氨酸34]GLP-1-(7-37)-肽[Q-linker-f8-9, arginine 34] GLP-1-(7-37)-peptide

步驟1step 1

Fmoc-Rink醯胺-MBHA樹脂Fmoc-Rink guanamine-MBHA resin

MS實測值：1127(M+3H)³⁺ ，MS理論值：1127(M+3H)³⁺ 。MS found: 1127 (M+3H) ³⁺ , MS: 1127 (M+3H) ³⁺ .

實施例7Example 7

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-c8, arginine 34]GLP-1-(7-37)-peptide

將[A-linker-c8,Arg34]GLP-1-OH(36mg,11μmol)、EDPA(4.0mg,30.8μmol)、乙腈(260μl)和水(260μl)的混合物在室溫下緩慢地攪拌5min。然後加入含有N^α -十六醯基-谷氨酸N-羥基琥珀醯亞胺酯-叔丁醚(1.8mg,3.3μmol)的乙腈(44.2μl)溶液，在室溫下緩慢攪拌混合反應1小時20分鐘後，加入溶於50%含水乙醇的甘氨酸(1.8mg,242μmol)溶液終止反應。再加入0.5%的含醋酸銨(12ml)和NMP(300μl)的水溶液，所得混合物上Varian 1g C₈ Mega Bondcartridge柱芯洗脫，用5%的乙腈(10ml)洗脫固定化的化合物，最後用TFA(6ml)洗脫使樣品從柱芯上解吸附。所得洗脫液在室溫下放置2小時，然後真空濃縮。所得殘留物質用柱色譜法和標準的乙腈，分離獲得目標化合物(12mg,46%)，用PDMS分析。質子化分子離子m/z實測值：3790±3。因此，結果分子量為3790±3 amu(理論值3751 amu)。A mixture of [A-linker-c8, Arg34] GLP-1-OH (36 mg, 11 μmol), EDPA (4.0 mg, 30.8 μmol), acetonitrile (260 μl) and water (260 μl) was slowly stirred at room temperature for 5 min. Then, a solution of N ^α -hexadecanyl-glutamic acid N-hydroxysuccinimide-tert-butyl ether (1.8 mg, 3.3 μmol) in acetonitrile (44.2 μl) was added, and the mixture was slowly stirred at room temperature. After 20 minutes in an hour, the reaction was stopped by adding a solution of glycine (1.8 mg, 242 μmol) dissolved in 50% aqueous ethanol. An additional 0.5% aqueous solution containing ammonium acetate (12 ml) and NMP (300 μl) was added to the resulting mixture on Varian 1g C ₈ Mega Bond The cartridge was eluted, the immobilized compound was eluted with 5% acetonitrile (10 ml), and finally eluted with TFA (6 ml) to desorb the sample from the core. The resulting eluate was allowed to stand at room temperature for 2 hours and then concentrated in vacuo. The residue obtained was purified by column chromatography eluting with EtOAc (EtOAc) Protonated molecular ion m / z measured value: 3790 ± 3. Therefore, the molecular weight was 3790 ± 3 amu (theoretical value 3751 amu).

實施例8Example 8

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34]GLP-1-(7-37)-peptide

合成目標GLP-1類似物使用與實施例7描述的相同序列和相同條件。The same sequence and the same conditions as described in Example 7 were used to synthesize the target GLP-1 analog.

MS實測值：1268(M+3H)³⁺ ，MS理論值：1268(M+3H)³⁺ 。MS found: 1268 (M+3H) ³⁺ , MS: 1268 (M+3H) ³⁺ .

實施例9Example 9

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-e8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-e8, arginine 34]GLP-1-(7-37)-peptide

MS實測值：1250(M+3H)³⁺ ，MS理論值：1250(M+3H)³⁺ 。MS found: 1250 (M+3H) ³⁺ , MS mp: 1250 (M+3H) ³⁺ .

實施例10Example 10

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-f8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-f8, arginine 34]GLP-1-(7-37)-peptide

MS實測值：1256(M+3H)³⁺ ，MS理論值：1256(M+3H)³⁺ 。MS found: 1256 (M+3H) ³⁺ , MS: 1256 (M+3H) ³⁺ .

實施例11Example 11

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-a8-9,精氨酸34]GLP-1-(7-37)-肽；N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-a8-9,arginine 34]GLP-1-(7-37) -peptide;

MS實測值：1244(M+3H)³⁺ ，理論MS：1244(M+3H)³⁺ 。MS found: 1244 (M+3H) ³⁺ , theory MS: 1244 (M+3H) ³⁺ .

實施例12Example 12

合成synthesis

N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-b8-9,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-b8-9,arginine 34]GLP-1-(7-37) -peptide

實施例13Example 13

合成synthesis

N-ε²⁶ -[(N^ε -ω-羧基十七醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[(N ^ε -ω-carboxyheptadecanyl)]-[Q-linker-c8,arginine 34]GLP-1-(7-37)-peptide

合成目標GLP-1類似物使用與實施例7描述的相同序列和相同條件，並使用ω-羧基十七醯基酸2,5-二氧吡咯烷-1-基酯作為起始原料替換N^α -十六醯基-谷氨酸N-羥基琥珀醯亞胺酯-叔丁醚。Synthesis of the target GLP-1 analog using the same sequence and the same conditions as described in Example 7, and replacing the ^{α α} with ω-carboxyheptadecanoic acid 2,5-dioxapyrrolidin-1-yl ester as a starting material - hexadecanolyl-glutamic acid N-hydroxysuccinimide-tert-butyl ether.

MS實測值：1239(M+3H)³⁺ ，MS理論值：1239(M+3H)³⁺ 。MS found: 1239 (M+3H) ³⁺ , MS mp.: 239 (M+3H) ³⁺ .

實施例14Example 14

合成synthesis

N-ε²⁶ -[(N^ε -ω-羧基十九醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽N-ε ²⁶ -[(N ^ε -ω-carboxynonanyl)]-[Q-linker-c8,arginine 34]GLP-1-(7-37)-peptide

合成目標GLP-1類似物使用與實施例7描述的相同序列和相同條件，並用ω-羧基十七醯基酸2,5-二氧吡咯烷-1-基酯作為起始原料替換N^α -十六醯基-谷氨酸N-羥基琥珀醯亞胺酯-叔丁醚。The synthetic target GLP-1 analogue was prepared using the same sequence and the same conditions as described in Example 7, and replacing N ^α with ω-carboxyheptadecanoic acid 2,5-dioxapyrrolidin-1-yl ester as a starting material. Hexadecyl-glutamic acid N-hydroxysuccinimide-tert-butyl ether.

MS實測值：1249(M+3H)³⁺ ，MS理論值：1249(M+3H)³⁺ 。MS found: 1249 (M+3H) ³⁺ , MS calc.: 1249 (M+3H) ³⁺ .

實施例15Example 15

合成synthesis

[Q-連接子-d8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-d8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

將[A-連接子-d8]　GLP-1-(7-37)-半胱氨酸-丙氨酸-NH₂ (36mg,11μmol)溶解到50mmol/L的Tris-HCl緩衝液(36mL)中得混合物，在室溫下與2mol過量的20KDa mPEG-SPA(用50 mmol/L Tris-HCl緩衝液調節將pH從7.5調至9.0)反應3小時。在室溫條件下，用X-tera C₁₈ (4.6 X 250 mm,5μm,Waters,Milford,MA)反相高效液相色譜(RP-HPLC)對單聚乙二醇GLP-1交聯物進行檢測和純化。流動相包括：0.1% TFA的水溶液(A相)，0.1% TFA的乙腈溶液(B相)。該流動相洗脫的線性梯度為30-60% B相，流速為1mL/min，洗脫時間為20min，在波長215nm處檢測UV吸光度。分別收集各峰值對應的HPLC組分，用氮氣淨化，凍乾。[A-linker-d8] GLP-1-(7-37)-cysteine-alanine-NH ₂ (36 mg, 11 μmol) was dissolved in 50 mmol/L Tris-HCl buffer (36 mL) The mixture was reacted for 3 hours at room temperature with 2 mol excess of 20 KDa mPEG-SPA (adjusted from 7.5 to 9.0 with 50 mmol/L Tris-HCl buffer). Monoethylene glycol GLP-1 cross-linking was carried out by reverse-phase high performance liquid chromatography (RP-HPLC) with X-tera C ₁₈ (4.6 X 250 mm, 5 μm, Waters, Milford, MA) at room temperature. Detection and purification. The mobile phase consisted of an aqueous solution of 0.1% TFA (phase A) and a 0.1% solution of TFA in acetonitrile (phase B). The mobile phase eluted with a linear gradient of 30-60% B phase, a flow rate of 1 mL/min, an elution time of 20 min, and UV absorbance at a wavelength of 215 nm. The HPLC components corresponding to the respective peaks were separately collected, purified with nitrogen, and lyophilized.

實施例16Example 16

合成synthesis

[Q-連接子-c8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-c8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

合成目標GLP-1類似物使用與實施例13描述的相同序列和相同條件，並使用20KDa mPEG-SPA。The synthetic target GLP-1 analog was used in the same sequence and the same conditions as described in Example 13, and 20KDa mPEG-SPA was used.

實施例17Example 17

合成synthesis

[Q-連接子-a8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-a8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

實施例18Example 18

合成synthesis

[Q-連接子-b8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-b8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

實施例19Example 19

合成synthesis

[Q-連接子-e8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-e8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

實施例20Example 20

合成synthesis

[Q-連接子-f8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ [Q-linker-f8]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂

合合成目標GLP-1類似物使用與實施例13描述的相同序列和相同條件，並使用20KDa mPEG-SPA。The synthetic target GLP-1 analog was synthesized using the same sequence and the same conditions as described in Example 13, and 20KDa mPEG-SPA was used.

實施例21Example 21

DPP-IV的體外穩定性In vitro stability of DPP-IV

將等量的本公司內部純化合成的GLP-1類似物(100μL,5nmol/L)，用鹽酸三乙基胺緩衝液(10mmol/L；pH7.4)預處理。然後加入DPP-IV(5mU,900μL)，所得溶液在溫度37℃下孵育。在各個孵育時間點，從反應體系中取出100μL反應液，然後加入5μL的10%(v/v)TFA終止反應。所得各份樣品用所描述的MALDI-TOF MS和RP-HPLC進行分析。An equal amount of the internal purified GLP-1 analog (100 μL, 5 nmol/L) was pretreated with triethylamine hydrochloride buffer (10 mmol/L; pH 7.4). Then DPP-IV (5 mU, 900 μL) was added and the resulting solution was incubated at 37 °C. At each incubation time point, 100 μL of the reaction solution was taken out from the reaction system, and then 5 μL of 10% (v/v) TFA was added to terminate the reaction. Each of the obtained samples was analyzed by the described MALDI-TOF MS and RP-HPLC.

實施例22Example 22

在表達克隆的人GLP-1受體的細胞系中的cAMP形成為了證明GLP-1衍生物的功效，測試GLP-1衍生物刺激表達克隆的人GLP-1受體的細胞系中cAMP形成的能力。採用劑量反應曲線計算EC₅₀ 。cAMP formation in a cell line expressing a cloned human GLP-1 receptor To demonstrate the efficacy of a GLP-1 derivative, the GLP-1 derivative was tested to stimulate cAMP formation in a cell line expressing a cloned human GLP-1 receptor. ability. EC ₅₀ is calculated using the dose-response curves.

在該放射免疫法中，使用NIT-1細胞，一種建立自轉基因NOD/Lt小鼠的胰島β-細胞系。該試驗在96-微孔板中開展，總體積140μL。使用的緩衝液是50 mmol/L Tris-HCl，pH 7.4，添加有1mmol/LEGTA、1.5mmol/L MgSO₄ 、1.7mmol/L ATP、20 mM GTP、2mmol/L 3-異丁基-1-甲基黃嘌呤、0.01%吐溫-20和0.1%人血清白蛋白。將用於激動劑活性測試的化合物在緩衝液中溶解和稀釋後，進行膜製備。所得混合液在37℃下孵育2h。加入25μl 0.05mol/L HCl終止反應。樣品稀釋10倍後，採用鄰近閃爍分析法分析cAMP。In this radioimmunoassay, NIT-1 cells, an islet β-cell line that establishes a transgenic NOD/Lt mouse, are used. The assay was performed in a 96-well microplate with a total volume of 140 μL. The buffer used was 50 mmol/L Tris-HCl, pH 7.4, with 1 mmol/LEGTA, 1.5 mmol/L MgSO ₄ , 1.7 mmol/L ATP, 20 mM GTP, 2 mmol/L 3-isobutyl-1- Methylxanthine, 0.01% Tween-20 and 0.1% human serum albumin. After the compound for the agonist activity test was dissolved and diluted in a buffer, membrane preparation was carried out. The resulting mixture was incubated at 37 ° C for 2 h. The reaction was stopped by adding 25 μl of 0.05 mol/L HCl. After the sample was diluted 10 times, cAMP was analyzed by proximity scintillation analysis.

實施例23Example 23

GLP-1類似物的抗高血糖活性Antihyperglycemic activity of GLP-1 analogues

將自發性糖尿病db/db小鼠分成4組(n=5)，禁食16小時。Spontaneous diabetic db/db mice were divided into 4 groups (n=5) and fasted for 16 hours.

然後向小鼠腹腔內注射生理鹽水、劑量100 μg/kg的GLP-1(7-36)醯胺、劑量100 μg eq/kg的[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽和劑量100 μg eq/kg的N-ε²⁶ -[γ-L-谷醯基(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽，注射10 min後灌胃葡萄糖溶液負荷，劑量為1 g/kg。The mice were then intraperitoneally injected with normal saline, 100 μg/kg GLP-1 (7-36) guanamine, and 100 μg eq/kg of [Q-linker-c8, glutamic acid 22] GLP-1. -(7-37)-peptide and dose of 100 μg eq/kg of N-ε ²⁶ -[γ-L-glutenyl (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34] GLP-1-(7-37)-peptide, administered with glucose solution at a dose of 1 g/kg after 10 min of injection.

於糖負荷-10min、0min、10min、20min、30min、60min、90min、120min和180min時，採集血樣並測試血糖水準。Blood samples were taken and tested for blood glucose levels at -10 min, 0 min, 10 min, 20 min, 30 min, 60 min, 90 min, 120 min, and 180 min.

通過計算0-180min內血糖水平-時間曲線下的面積，比較GLP-1(7-36)醯胺、[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽和N-ε²⁶ -[γ-L-谷醯胺(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽抑制血糖升高的效果。Comparison of GLP-1 (7-36) guanamine, [Q-linker-c8, glutamic acid 22] GLP-1-(7-37)- by calculating the area under the blood glucose level-time curve within 0-180 min Peptide and N-ε ²⁶ -[γ-L-glutamine (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34]GLP-1-(7-37)- The peptide inhibits the effect of elevated blood sugar.

GLP-1(7-36)醯胺組的AUC值是25165±4463mg-min/dl，相對生理鹽水組的AUC值(34864+4774 mg.min/dl)下降了27.8%。The AUC value of the GLP-1(7-36) indole group was 25165±4463 mg-min/dl, which was 27.8% lower than the AUC value of the saline group (34864+4774 mg.min/dl).

然而，[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽和N-ε²⁶ -[γ-L-谷醯基(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽的AUC值分別是14470+5700mg-min/dl和17520+2484 mg-min/dl(分別下降了58.5%和49.7%)。However, [Q-linker-c8, glutamic acid 22] GLP-1-(7-37)-peptide and N-ε ²⁶ -[γ-L-glutenyl (N-α-hexadecanyl)]- [Q-linker-d8, arginine 34] The AUC values of GLP-1-(7-37)-peptide were 14470+5700 mg-min/dl and 17520+2484 mg-min/dl, respectively (down 58.5, respectively). % and 49.7%).

這些結果表明：同GLP-I(7-36)醯胺相比，[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽和N-ε²⁶ -[γ-L-谷醯胺酸(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽具有顯著地增強的抑制血糖水平活性。These results indicate that [Q-linker-c8, glutamic acid 22] GLP-1-(7-37)-peptide and N-ε ²⁶ -[γ compared to GLP-I(7-36) decylamine -L-glutamyric acid (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34] GLP-1-(7-37)-peptide has a markedly enhanced inhibition of blood sugar Horizontal activity.

綜上所述，本發明確實可達到上述諸項功能及目的，故本發明應符合專利申請要件，爰依法提出申請。In summary, the present invention can achieve the above functions and purposes, so the present invention should meet the requirements of the patent application, and apply in accordance with the law.

Claims

一種式(I)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽，Xaa₇ -Q-Gly-Thr-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Tyr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B (I)其特徵在於：Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β -羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Q選自下述連接子(II)、(III)或(IV)：式中，R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z選自N、O或S時，W不存在；當Z是C時，W選自H或F； Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ -谷氨酸、β -丙氨酸、γ -氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ -谷氨酸、β -丙氨酸或γ -氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸，當Q是連接子(II)或(IV)時，B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段；當Q是連接子(III)時，B選自2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue represented by the formula (I), a composition thereof or a pharmaceutically acceptable salt thereof, Xaa ₇ -Q-Gly-Thr-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Tyr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B (I) It is: Xaa ₇ is natural or non-natural, selected from L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β -hydroxy-histidine, high An amino acid of histidine, α-fluoromethyl-histidine or α-methyl-histidine; Q is selected from the following linkers (II), (III) or (IV): Wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; and X is selected from H, F, OH, CF ₃ or O; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is selected from N, O or S, W is not Exist; when Z is C, W is selected from H or F; Xaa ₁₄ is natural or non-natural, amino acid selected from serine or histidine; Xaa ₁₆ is natural or non-natural, selected from valine, Lai Amino acid of leucine or leucine; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from serine, arginine or lysine; Xaa ₂₂ is natural or non- A natural amino acid selected from the group consisting of glycine, aminoisobutyric acid or glutamic acid; Xaa ₂₃ is glutamine; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural, selected from glycine An amino acid of lysine, arginine, leucine, aspartame or aminoisobutyric acid; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ -glutamic acid, β -alanine Acid, γ -aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7 , 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Only T is an amino acid selected from γ -glutamic acid, β -alanine or γ -aminobutyric acid, or T is selected from Where is present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids, when Q is a linker (II) or (IV), B is selected from glycine, glycosamine or from 2 to 5 natural or unnatural amino acids and must have one The amino acid is a peptide fragment consisting of cysteine; when Q is a linker (III), B is selected from 2 to 5 natural or unnatural amino acids and must have a peptide fragment consisting of a cysteine.

如申請專利範圍第1項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The GLP-1 analogue, composition or pharmaceutically acceptable salt thereof according to claim 1, wherein Xaa ₂₆ is natural or non-natural, selected from the group consisting of glycine, lysine, and fine Amino acid, leucine and aspartame or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

如申請專利範圍第2項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：所述肽片段為半胱氨酸-絲氨酸-甘氨酸、半胱氨酸-丙氨酸或連接著半胱氨酸的甲氧基聚乙二醇馬來醯亞胺。 A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof according to claim 2, wherein the peptide fragment is cysteine-serine-glycine or cysteine - Alanine or methoxypolyethylene glycol maleimide linked to cysteine.

一種式(V)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽，式中， R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H；X選自H、F、OH、CF₃ 或O；D是Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β -羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ -谷氨酸、β -丙氨酸、γ -氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ -谷氨酸、β -丙氨酸或γ -氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；當Q是連接子(II)或(IV)時，B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段；當Q是連接子(III)時，B選自由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue represented by the formula (V), a composition thereof, or a pharmaceutically acceptable salt thereof, Wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; and X is selected from H, F, OH, CF ₃ or O; D is Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe- Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa ₇ is natural or non-natural, selected from L-histidine, D-histidine, deamination-histamine Amino acid of acid, 2-amino-histidine, β -hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine; Xaa ₁₄ is natural or non- a natural amino acid selected from serine or histidine; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine; Xaa ₂₂ is a natural or non-natural amino acid selected from glycine, aminoisobutyric acid or glutamic acid; Xaa ₂₃ is Glutamine; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural, selected from glycine, lysine, arginine An amino acid of leucine, aspartame or aminoisobutyric acid; or Xaa ₂₆ is a lysine linked to TU; T is selected from γ -glutamic acid, β -alanine, γ -aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, _{16, 17} , 18, ₁₉ , 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ -glutamic acid, β -alanine or γ -aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids; when Q is a linker (II) or (IV), B is selected from glycine, glycine or from 2 to 5 natural or unnatural amino acids and must have one The amino acid is a peptide fragment consisting of cysteine; when Q is a linker (III), B is selected from peptide fragments consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

如申請專利範圍第4項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 4, wherein Xaa ₂₆ is natural or non-natural, selected from the group consisting of glycine, lysine, and fine Amino acid, leucine and aspartame or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

如申請專利範圍第4項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：所述肽片段為半胱氨酸-絲氨酸-甘氨酸、半胱氨酸-丙氨酸或連接著半胱氨酸的甲氧基聚乙二醇馬來醯亞胺。 A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 4, wherein the peptide fragment is cysteine-serine-glycine, cysteine - Alanine or methoxypolyethylene glycol maleimide linked to cysteine.

如申請專利範圍第4項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：R₁ 和R₂ 均選自H或CH₃ 且R₃ 是H，或者R₁ 是CH₃ ，R₂ 和R₃ 均是H；或者R₁ 和R₃ 均是H，R₂ 是CH₃ 。The GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 4, wherein R ₁ and R ₂ are each selected from H or CH ₃ and R ₃ is H, Or R ₁ is CH ₃ , R ₂ and R ₃ are both H; or both R ₁ and R ₃ are H and R ₂ is CH ₃ .

一種式(VI)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H；Y選自H、OH、F或(C₁ -C₆ )烷基；D是+Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa7是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；或者Xaa₁₆ 是連接著T-U的賴氨酸； Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸或γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；B選自由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue represented by the formula (VI), a composition thereof, or a pharmaceutically acceptable salt thereof, Characterized in that R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; Y is selected from H, OH, F Or (C ₁ -C ₆ )alkyl; D is +Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa _26- Xaa ₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa7 is natural or non-natural, selected from L-histidine, D-histidine An amino acid of deamination-histidine, 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl-histidine; Xaa ₁₄ is a natural or non-natural amino acid selected from serine or histidine; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of valine, lysine or leucine; or Xaa ₁₆ is linked Lysine of TU; Xaa ₁₈ is a natural or non-natural amino acid selected from serine, arginine or lysine; Xaa ₂₂ is natural or non-natural, selected from glycine, aminoisobutyric acid or glutamic acid amino acid; Xaa ₂₃ is _{_{glutamine; Xaa 26, Xaa 27, Xaa}} 34, Xaa 35 and Xaa ₃₆ are natural or unnatural, selected from glycine, lysine Acid, arginine, leucine, amino isobutyric acid or asparagine amino; or Xaa ₂₆ is connected to a TU-lysine; glutamic acid γ- T is selected from, [beta] -alanine, gamma] - aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, ₁₆ 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine or γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids; B is selected from peptide fragments consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

如申請專利範圍第8項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 8, wherein Xaa ₂₆ is natural or non-natural, selected from the group consisting of glycine, lysine, and fine Amino acid, leucine and aspartame or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

如申請專利範圍第8項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：所述肽片段為半胱氨酸-絲氨酸-甘氨酸、半胱氨酸-丙氨酸或連接著半胱氨酸的甲氧基聚乙二醇馬來醯亞胺。 A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 8, wherein the peptide fragment is cysteine-serine-glycine, cysteine - Alanine or methoxypolyethylene glycol maleimide linked to cysteine.

如申請專利範圍第8項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基；R₃ 是H；Y選自H或(C₁ -C₆ )烷基。A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 8, wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R ₂ It is selected from H or (C ₁ -C ₆ )alkyl; R ₃ is H; and Y is selected from H or (C ₁ -C ₆ )alkyl.

一種式(VII)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽，式中，R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H； Y選自H、OH、F或(C₁ -C₆ )烷基；W選自H或F；D是Gly-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Thr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -B；Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；或者Xaa₂₆ 是連接著T-U的賴氨酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸；B選自甘氨酸、甘氨醯胺或由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。A GLP-1 analogue represented by the formula (VII), a composition thereof, or a pharmaceutically acceptable salt thereof, Wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; W is selected from H or F; D is Gly-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Thr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ - Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -B; Xaa ₇ is natural or non-natural, selected from L-histidine , D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, homohistidine, α-fluoromethyl-histidine or α-methyl- An amino acid of histidine; Xaa ₁₄ is a natural or non-natural amino acid selected from serine or histidine; Xaa ₁₆ is a natural or non-natural amino acid selected from the group consisting of proline, lysine or leucine; Or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine; Xaa ₂₂ is natural or non-natural, selected from glycine and amino amino acid or glutamic acid; Xaa ₂₃ is _{_{glutamine; Xaa 26, Xaa 27, Xaa}} 34, Xaa 35 and Xaa ₃₆ are natural or unnatural, selected from glycine Lysine, arginine, leucine, amino isobutyric acid or asparagine amino; or Xaa ₂₆ is connected to a TU-lysine; glutamic acid γ- T is selected from, [beta] -alanine , γ-aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27; or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Only T is an amino acid selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids; B is selected from glycine, glycosamine or a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and having one amino acid being cysteine.

如申請專利範圍第12項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：Xaa₂₆ 是天然或非天然的、選自甘氨酸、賴氨酸、精氨酸、亮氨酸和天冬醯胺或沒有連接T-U的α-氨基異丁酸，B為由2~5個天然或非天然氨基酸且必須有一個氨基酸是半胱氨酸組成的肽片段。The GLP-1 analogue, composition or pharmaceutically acceptable salt thereof according to claim 12, wherein the Xaa ₂₆ is natural or non-natural, selected from the group consisting of glycine, lysine, and fine Amino acid, leucine and aspartame or α-aminoisobutyric acid without TU, B is a peptide fragment consisting of 2 to 5 natural or unnatural amino acids and one amino acid is cysteine.

如申請專利範圍第12項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：所述肽片段為半胱氨酸-絲氨酸-甘氨酸、半胱氨酸-丙氨酸或連接著半胱氨酸的甲氧基聚乙二醇馬來醯亞胺。 The GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, according to claim 12, wherein the peptide fragment is cysteine-serine-glycine, cysteine - Alanine or methoxypolyethylene glycol maleimide linked to cysteine.

如申請專利範圍第12項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽，其特徵在於：R₃ 是H；Y是H或F。The GLP-1 analogue, composition or pharmaceutically acceptable salt thereof according to claim 12, wherein R ₃ is H; Y is H or F.

一種式(VIII)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽， Xaa7是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β-羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Q選自下述連接子(II)、(III)或(IV)：式中，R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z是N、O或S時，W不存在；當Z是C時，W選自H或F；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸； Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；T選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸或HOOC(CH₂ )_n COOH，其中，n選自1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26或27；或者T選自，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U僅僅在T為選自γ-谷氨酸、β-丙氨酸、γ-氨基丁酸的氨基酸，或者T選自時存在，式中，k選自1、2、3、4、5、6、7、8、9或10，m選自1、2、3、4、5、6、7、8、9或10；U選自C₈ -C₂₀ 脂肪酸，B選自甘氨酸或甘氨醯胺。A GLP-1 analogue represented by the formula (VIII), a composition thereof, or a pharmaceutically acceptable salt thereof, Xaa7 is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β-hydroxy-histidine, homohistidine An amino acid of α-fluoromethyl-histidine or α-methyl-histidine; Q is selected from the following linkers (II), (III) or (IV): Wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; and X is selected from H, F, OH, CF ₃ or O; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is N, O or S, W does not exist When Z is C, W is selected from H or F; Xaa ₁₄ is natural or non-natural, amino acid selected from serine or histidine; Xaa ₁₆ is natural or non-natural, selected from valine and lysine An amino acid of acid or leucine; Xaa ₁₈ is a natural or non-natural amino acid selected from the group consisting of serine, arginine or lysine; Xaa ₂₂ is natural or non-natural, selected from glycine, aminoisobutyric acid or valley Amino acid of amino acid; Xaa ₂₃ is glutamine; Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural, selected from glycine, lysine, arginine, leucine, aspartame An amino acid of an amine or aminoisobutyric acid; T is selected from the group consisting of γ-glutamic acid, β-alanine, γ-aminobutyric acid or HOOC(CH ₂ ) _n COOH, wherein n is selected from 1, 2, 3, 4 , 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 Or T is selected from Wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; U is only an amino acid selected from γ-glutamic acid, β-alanine, γ-aminobutyric acid at T, or T is selected from T When present, wherein k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; U is selected from C ₈ -C ₂₀ fatty acids, and B is selected from glycine or glycine.

一種式(IX)表示的GLP-1類似物、其組合物或其藥學上可接受的鹽，Xaa₇ -Q-Gly-Thr-Phe-Thr-Xaa₁₄ -Asp-Xaa₁₆ -Ser-Xaa₁₈ -Tyr-Leu-Glu-Xaa₂₂ -Xaa₂₃ -Ala-Ala-Xaa₂₆ -Xaa₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa₃₄ -Xaa₃₅ -Xaa₃₆ -Gly-Xaaⁿ -Xaaⁿ⁺¹ -Xaaⁿ⁺² -Xaaⁿ⁺³ -Xaaⁿ⁺⁴ -Cys^(PEG) -Xaa^m -Xaa^m+1 -Xaa^m+2 -Xaa^m+3 -Xaa^m+4 IX式中：Xaa₇ 是天然的或非天然的，選自L-組氨酸、D-組氨酸、脫氨基-組氨酸、2-氨基-組氨酸、β -羥基-組氨酸、高組氨酸、α-氟甲基-組氨酸或α-甲基-組氨酸的氨基酸；Q選自下述連接子(II)、(III)和(IV)：式中，R₁ 選自H或(C₁ -C₆ )烷基；R₂ 選自H或(C₁ -C₆ )烷基或；R₃ 為H；X選自H、F、OH、CF₃ 或O；Y選自H、OH、F或(C₁ -C₆ )烷基；Z選自N、C、O或S；其中，當Z是N、O或S時，W不存在；當Z是C時，W選自H或F；Xaa₁₄ 是天然或非天然的，選自絲氨酸或組氨酸的氨基酸；Xaa₁₆ 是天然或非天然的，選自纈氨酸、賴氨酸或亮氨酸的氨基酸；或者Xaa₁₆ 是連接著T-U的賴氨酸；Xaa₁₈ 是天然或非天然的，選自絲氨酸、精氨酸或賴氨酸的氨基酸；Xaa₂₂ 是天然或非天然的，選自甘氨酸、氨基異丁酸或谷氨酸的氨基酸；Xaa₂₃ 是谷氨醯胺；Xaa₂₆ 、Xaa₂₇ 、Xaa₃₄ 、Xaa₃₅ 和Xaa₃₆ 是天然或非天然的，選自甘氨酸、賴氨酸、精氨酸、亮氨酸、天冬醯胺或氨基異丁酸的氨基酸；或者Xaa26是連接著T-U的賴氨酸；Xaaⁿ 、Xaaⁿ⁺¹ 、Xaaⁿ⁺² 、Xaaⁿ⁺³ 、Xaaⁿ⁺⁴ 、Xaa^m 、Xaa^m+1 、Xaa^m+2 、Xaa^m+3 及Xaa^m+4 一起組成1、2、3或4個天然的或非天然的氨基酸；也就是說，Xaaⁿ 、Xaaⁿ⁺¹ 、Xaaⁿ⁺² 、Xaaⁿ⁺³ 、Xaaⁿ⁺⁴ 、Xaa^m 、Xaa^m+1 、Xaa^m+2 、Xaa^m+3 和Xaa^m+4 與連接著甲氧基聚乙二醇馬來醯亞胺的半胱氨酸形成2~5個氨基酸的片段。A GLP-1 analogue represented by the formula (IX), a composition thereof or a pharmaceutically acceptable salt thereof, Xaa ₇ -Q-Gly-Thr-Phe-Thr-Xaa ₁₄ -Asp-Xaa ₁₆ -Ser-Xaa ₁₈ -Tyr-Leu-Glu-Xaa ₂₂ -Xaa ₂₃ -Ala-Ala-Xaa ₂₆ -Xaa ₂₇ -Phe-Ile-Ala-Trp-Leu-Val-Xaa ₃₄ -Xaa ₃₅ -Xaa ₃₆ -Gly-Xaa ⁿ -Xaa ⁿ⁺¹ -Xaa ⁿ⁺² -Xaa ⁿ⁺³ -Xaa ⁿ⁺⁴ -Cys ^(PEG) -Xaa ^m -Xaa ^m+1 -Xaa ^m+2 -Xaa ^m+3 -Xaa ^m+4 IX where: Xaa ₇ is natural or non-natural, selected from the group consisting of L-histidine, D-histidine, dea-histidine, 2-amino-histidine, β -hydroxy-histidine, high histamine An amino acid of acid, α-fluoromethyl-histidine or α-methyl-histidine; Q is selected from the following linkers (II), (III) and (IV): Wherein R _{1 is} selected from H or (C ₁ -C ₆ )alkyl; R _{2 is} selected from H or (C ₁ -C ₆ )alkyl or; R ₃ is H; and X is selected from H, F, OH, CF ₃ or O; Y is selected from H, OH, F or (C ₁ -C ₆ )alkyl; Z is selected from N, C, O or S; wherein, when Z is N, O or S, W does not exist When Z is C, W is selected from H or F; Xaa ₁₄ is natural or non-natural, amino acid selected from serine or histidine; Xaa ₁₆ is natural or non-natural, selected from valine and lysine Acid or leucine amino acid; or Xaa ₁₆ is a lysine linked to TU; Xaa ₁₈ is a natural or non-natural amino acid selected from serine, arginine or lysine; Xaa ₂₂ is natural or unnatural An amino acid selected from the group consisting of glycine, aminoisobutyric acid or glutamic acid; Xaa ₂₃ is glutamine; Xaa ₂₆ , Xaa ₂₇ , Xaa ₃₄ , Xaa ₃₅ and Xaa ₃₆ are natural or non-natural, selected from glycine, An amino acid of lysine, arginine, leucine, aspartame or aminoisobutyric acid; or Xaa26 is a lysine linked to TU; Xaa ⁿ , Xaa ⁿ⁺¹ , Xaa ⁿ⁺² , Xaa ^{n +3} , Xaa ⁿ⁺⁴ , Xaa ^m , Xaa ^m+1 , Xaa ^m+2 , Xaa ^m+3 and Xaa ^m+4 together form 1, 2, 3 or 4 a natural or non-natural amino acid; that is, Xaa ⁿ , Xaa ⁿ⁺¹ , Xaa ⁿ⁺² , Xaa ⁿ⁺³ , Xaa ⁿ⁺⁴ , Xaa ^m , Xaa ^m+1 , Xaa ^m+2 , Xaa ^m+3 and Xaa ^m+4 form a fragment of 2 to 5 amino acids with a cysteine linked to methoxypolyethylene glycol maleimide.

如申請專利範圍第1-17項中任一項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽選自：[Q-連接子-d8,谷氨酸22]GLP-1-(7-37)-肽；[Q-連接子-a8-9,谷氨酸22]GLP-1-(7-37)-肽；[Q-連接子-b8-9,谷氨酸22]GLP-1-(7-37)-肽；[Q-連接子-c8,谷氨酸22]GLP-1-(7-37)-肽； [Q-連接子-e8-9,谷氨酸22]GLP-1-(7-37)-肽；[Q-連接子-f8-9,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-d8,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-e8,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-f8,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ-L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-a8-9,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[γ -L-谷氨醯基(N-α-十六醯基)]-[Q-連接子-b8-9,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[(N^ε -ω-羧基十七醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；N-ε²⁶ -[(N^ε -ω-羧基十九醯基)]-[Q-連接子-c8,精氨酸34]GLP-1-(7-37)-肽；[Q-連接子-d8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；[Q-連接子-c8]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；[Q-連接子-a8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；[Q-連接子-b8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ； [Q-連接子-e8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ ；[Q-連接子-f8-9]GLP-1-(7-37)-半胱氨酸^(PEG) -氨基酮戊酸-NH₂ 。The GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-17, which is selected from the group consisting of: [Q-linker-d8, glutamic acid 22] GLP -1-(7-37)-peptide; [Q-linker-a8-9, glutamic acid 22] GLP-1-(7-37)-peptide; [Q-linker-b8-9, glutamine Acid 22] GLP-1-(7-37)-peptide; [Q-linker-c8, glutamic acid 22] GLP-1-(7-37)-peptide; [Q-linker-e8-9, Glutamate 22] GLP-1-(7-37)-peptide; [Q-linker-f8-9, arginine 34] GLP-1-(7-37)-peptide; N-ε ²⁶ -[ γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-c8, arginine 34]GLP-1-(7-37)-peptide; N-ε ²⁶ - [γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-d8, arginine 34] GLP-1-(7-37)-peptide; N-ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-e8, arginine 34]GLP-1-(7-37)-peptide; N-ε ^26- [γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-f8, arginine 34]GLP-1-(7-37)-peptide; N- ε ²⁶ -[γ-L-glutamyl (N-α-hexadecanyl)]-[Q-linker-a8-9, arginine 34]GLP-1-(7-37)-peptide ;N-ε ²⁶ -[ γ -L-glutamyl (N-α-hexadecanyl)]-[Q-linker-b8-9, arginine 34]GLP-1-(7-37 )-peptide; N-ε ²⁶ -[(N ^Ε -ω-carboxyheptadecanyl)]-[Q-linker-c8,arginine 34]GLP-1-(7-37)-peptide; N-ε ²⁶ -[(N ^ε -ω-carboxyl 19-mercapto)]-[Q-linker-c8, arginine 34]GLP-1-(7-37)-peptide; [Q-linker-d8]GLP-1-(7-37)- Cysteine ^(PEG) -aminolevulinic acid-NH ₂ ; [Q-linker-c8] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ; [Q-linker-a8-9]GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ;[Q-linker-b8-9]GLP-1- (7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ ; [Q-linker-e8-9]GLP-1-(7-37)-cysteine ^(PEG) - Aminolevulinic acid-NH ₂ ; [Q-linker-f8-9] GLP-1-(7-37)-cysteine ^(PEG) -aminolevulinic acid-NH ₂ .

一種藥物組合物，包含有如申請專利範圍第1-18中任一項所述的GLP-1類似物、其組合物或其藥學上可接受的鹽及一種藥學上可接受的輔料。 A pharmaceutical composition comprising the GLP-1 analogue according to any one of claims 1 to 18, a composition thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant.

如申請專利範圍第19項所述的藥物組合物，其特徵在於：該藥物組合物適用於胃腸用藥。 The pharmaceutical composition according to claim 19, wherein the pharmaceutical composition is suitable for gastrointestinal administration.

一種GLP-1類似物、其組合物或其藥學上可接受的鹽在製備治療或預防高血糖症、II型糖尿病、糖耐量受損、I型糖尿病、肥胖症、高血壓、X綜合症、血脂異常、認知障礙、動脈硬化、心肌梗塞、冠狀動脈心臟病、中風、炎性腸道綜合症、消化不良和胃潰瘍藥物、治療II型糖尿病的藥效延遲或預防惡化的藥物、以及用於減少食物攝入量、減少β細胞凋亡、增加胰島β細胞功能、增加β-細胞團和/或恢復葡萄糖對β-細胞的敏感性的藥物中的應用，其中所述GLP-1類似物是如申請專利範圍第1-18項中任一項所述GLP-1類似物。 A GLP-1 analogue, a composition thereof, or a pharmaceutically acceptable salt thereof, for the preparation or treatment of hyperglycemia, type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, syndrome X, Dyslipidemia, cognitive impairment, arteriosclerosis, myocardial infarction, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer drugs, drugs for delaying or preventing deterioration of type 2 diabetes, and for reducing Use of a food intake, a drug that reduces beta cell apoptosis, increases islet beta cell function, increases beta-cell mass, and/or restores glucose sensitivity to β-cells, wherein the GLP-1 analog is The GLP-1 analogue according to any one of claims 1 to 18.

一種藥物組合物在製備治療或預防高血糖症、II型糖尿病、糖耐量受損、I型糖尿病、肥胖症、高血壓、X綜合症、血脂異常、認知障礙、動脈硬化、心肌梗塞、冠狀動脈心臟病、中風、炎性腸道綜合症、消化不良和胃潰瘍藥物、治療II型糖尿病的藥效延遲或預防惡化的藥物、以及用於減少食物攝入量、減少β細胞凋亡、增加胰島β細胞功能、增加β-細胞團和/或恢復葡萄糖對β-細胞的敏感性的藥物中的應用，其中所述GLP-1類似物是如申請專利範圍第1-18項中任一項所述GLP-1類似物。A pharmaceutical composition for treating or preventing hyperglycemia, type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive impairment, arteriosclerosis, myocardial infarction, coronary artery Heart disease, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer drugs, drugs for delaying or preventing deterioration of type 2 diabetes, and for reducing food intake, reducing beta cell apoptosis, and increasing islet β Cell function, increase β-cell mass and/or restore glucose to β-fine The use of a cell for the sensitivity of a drug, wherein the GLP-1 analogue is a GLP-1 analogue as described in any one of claims 1-18.