JP2003261595A - Biotin-like peptide lipid - Google Patents
Biotin-like peptide lipidInfo
- Publication number
- JP2003261595A JP2003261595A JP2002060652A JP2002060652A JP2003261595A JP 2003261595 A JP2003261595 A JP 2003261595A JP 2002060652 A JP2002060652 A JP 2002060652A JP 2002060652 A JP2002060652 A JP 2002060652A JP 2003261595 A JP2003261595 A JP 2003261595A
- Authority
- JP
- Japan
- Prior art keywords
- gly
- brs
- mmol
- aminohexyl
- dodecyloxy
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 23
- 150000002632 lipids Chemical class 0.000 title claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 24
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 24
- 239000000463 material Substances 0.000 claims abstract description 12
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000004471 Glycine Substances 0.000 claims abstract description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims abstract description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims abstract description 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims abstract description 3
- 238000013268 sustained release Methods 0.000 claims description 18
- 239000012730 sustained-release form Substances 0.000 claims description 18
- XJFITURPHAKKAI-SRVKXCTJSA-N His-Pro-Gln Chemical group C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(N)=O)C(O)=O)C1=CN=CN1 XJFITURPHAKKAI-SRVKXCTJSA-N 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 230000005661 hydrophobic surface Effects 0.000 abstract description 6
- 238000004113 cell culture Methods 0.000 abstract description 5
- 239000011248 coating agent Substances 0.000 abstract description 4
- 238000000576 coating method Methods 0.000 abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 123
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 60
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 36
- 239000000243 solution Substances 0.000 description 28
- 238000006243 chemical reaction Methods 0.000 description 27
- 239000000203 mixture Substances 0.000 description 20
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 18
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 18
- 235000019341 magnesium sulphate Nutrition 0.000 description 18
- 238000005481 NMR spectroscopy Methods 0.000 description 17
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 16
- 238000000034 method Methods 0.000 description 15
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 12
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 10
- 239000002904 solvent Substances 0.000 description 10
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 9
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 9
- 108010090804 Streptavidin Proteins 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- 239000003102 growth factor Substances 0.000 description 7
- 230000003993 interaction Effects 0.000 description 7
- -1 lipid compound Chemical class 0.000 description 6
- 238000010898 silica gel chromatography Methods 0.000 description 6
- IOHPVZBSOKLVMN-UHFFFAOYSA-N 2-(2-phenylethyl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1CCC1=CC=CC=C1 IOHPVZBSOKLVMN-UHFFFAOYSA-N 0.000 description 5
- 108090001008 Avidin Proteins 0.000 description 5
- 230000010261 cell growth Effects 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 101800000021 N-terminal protease Proteins 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000000017 hydrogel Substances 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 description 3
- 229920005654 Sephadex Polymers 0.000 description 3
- 239000012507 Sephadex™ Substances 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- ZPGDWQNBZYOZTI-SFHVURJKSA-N (2s)-1-(9h-fluoren-9-ylmethoxycarbonyl)pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ZPGDWQNBZYOZTI-SFHVURJKSA-N 0.000 description 2
- WDGICUODAOGOMO-DHUJRADRSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-oxo-5-(tritylamino)pentanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)CC(=O)NC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 WDGICUODAOGOMO-DHUJRADRSA-N 0.000 description 2
- KXDAEFPNCMNJSK-UHFFFAOYSA-N Benzamide Chemical compound NC(=O)C1=CC=CC=C1 KXDAEFPNCMNJSK-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 125000004103 aminoalkyl group Chemical group 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 238000010511 deprotection reaction Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 230000005593 dissociations Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000009878 intermolecular interaction Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- XXMYDXUIZKNHDT-QNGWXLTQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(N=C1)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 XXMYDXUIZKNHDT-QNGWXLTQSA-N 0.000 description 1
- SDOWLLNYQHEAHC-UHFFFAOYSA-N 2-dodecoxybenzamide Chemical compound CCCCCCCCCCCCOC1=CC=CC=C1C(N)=O SDOWLLNYQHEAHC-UHFFFAOYSA-N 0.000 description 1
- QPBBFSXCZXADEC-UHFFFAOYSA-N 3,5-didodecoxybenzamide Chemical compound CCCCCCCCCCCCOC1=CC(OCCCCCCCCCCCC)=CC(C(N)=O)=C1 QPBBFSXCZXADEC-UHFFFAOYSA-N 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 235000018734 Sambucus australis Nutrition 0.000 description 1
- 244000180577 Sambucus australis Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000010976 amide bond formation reaction Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- RUDWJDABXVVFEN-UHFFFAOYSA-N n-(6-aminohexyl)-3,5-didodecoxybenzamide Chemical compound CCCCCCCCCCCCOC1=CC(OCCCCCCCCCCCC)=CC(C(=O)NCCCCCCN)=C1 RUDWJDABXVVFEN-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
(57)【要約】
【課題】本発明の目的は、疎水性表面にコーティングす
ることにより、細胞培養等に使用するタンパク質性の因
子を、材料表面から徐放する機能を付与する新規な構造
を有するビオチン様ペプチド脂質を提供することにあ
る。
【解決手段】
【化１】
式（１）で表されるペプチド脂質。
（式中Ｒ1は、Ｏ（ＣＨ2）nＣＨ3基を、Ｒ2はＨまたは
Ｒ1を表し、ｌは０から３の整数を、ｍは２から６の整
数を、ｎは１１から１７の整数を、Ｈｉｓはヒスチジン
を、Ｐｒｏはプロリンを、Ｇｌｎはグルタミンを、Ｇｌ
ｙはグリシンを表わす。）(57) [Summary] The object of the present invention is to provide a novel structure which, by coating a hydrophobic surface, imparts a function of gradually releasing a protein factor used for cell culture or the like from the material surface. Another object of the present invention is to provide a biotin-like peptide lipid having the same. SOLUTION: embedded image A peptide lipid represented by the formula (1). (Wherein R 1 represents an O (CH 2 ) n CH 3 group, R 2 represents H or R 1 , 1 is an integer of 0 to 3, m is an integer of 2 to 6, and n is 11 17 is an integer, His is histidine, Pro is proline, Gln is glutamine, Gl
y represents glycine. )
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、新規な構造を有す
るペプチド脂質化合物に関するものである。より詳細に
は、本発明は疎水性表面にコーティングすることによ
り、細胞培養等に使用するタンパク質性の因子を、材料
表面から徐放する機能を付与する新規な構造を有するビ
オチン様ペプチド脂質に関するものである。TECHNICAL FIELD The present invention relates to a peptide lipid compound having a novel structure. More specifically, the present invention relates to a biotin-like peptide lipid having a novel structure which imparts a function of gradually releasing a proteinaceous factor used for cell culture or the like from the material surface by coating the hydrophobic surface. Is.
【０００２】[0002]
【従来の技術】再生医療や人工臓器等の分野で、幹細胞
や体細胞の利用が盛んに研究されている。そうした細胞
の増殖、分化、活性化等の為に様々な増殖因子、分化誘
導因子、サイトカイン等のタンパク質性の因子が用いら
れている。例えば、細胞成長因子は細胞の増殖あるいは
分化を制御しているタンパク質であり、その作用やメカ
ニズムが明らかになるとともに、遺伝子操作により大量
生産が可能になっている。2. Description of the Related Art Utilization of stem cells and somatic cells has been actively studied in the fields of regenerative medicine and artificial organs. Various growth factors, differentiation-inducing factors, proteinaceous factors such as cytokines, etc. are used for the growth, differentiation, activation, etc. of such cells. For example, a cell growth factor is a protein that controls cell growth or differentiation, and its action and mechanism have been clarified, and mass production is possible by genetic engineering.
【０００３】そこで、これらを上手く利用すれば細胞の
増殖や分化を制御することが期待できる。しかしなが
ら、細胞成長因子を水溶液状態で投与するだけでは、期
待する生物効果は得られない。細胞成長因子を有効に利
用するためには、必要な濃度を、必要な期間にわたって
有効値に保つための工夫が必要である（田畑泰彦、化学
フロンティア３「再生医工学」、筏義人編、化学同人、
８１頁-９０頁、２００１年）。Therefore, if these are used effectively, it can be expected to control the proliferation and differentiation of cells. However, the expected biological effect cannot be obtained simply by administering the cell growth factor in an aqueous solution. In order to effectively use cell growth factors, it is necessary to devise to keep the required concentration at an effective value for the required period (Yasuhiko Tabata, Chemistry Frontier 3, "Regenerative Medicine Engineering", edited by Raito Yoshito, Chemistry). Coterie,
81-90, 2001).
【０００４】その現実的な一つの方法は細胞成長因子を
高分子キャリヤー内に含ませ、そこから持続的に徐放さ
せることである。例えば、水溶性タンパク質を徐放化す
る為に、油溶性の乳酸−グリコール酸共重合体と混合形
成する試みがあるが（Ｊ．Ｇａｏ等、Ｊ．Ｂｉｏｍｅ
ｄ．Ｍａｔｅｒ．Ｒｅｓ．、５５巻、５１２-５２２
頁、２００１年）、相溶性の悪い混合プロセスによって
タンパク質が変性し、その生物活性を失う場合が多い。One practical way to do this is to include the cell growth factor in a polymeric carrier from which sustained sustained release is achieved. For example, there is an attempt to form a mixture with an oil-soluble lactic acid-glycolic acid copolymer in order to gradually release a water-soluble protein (J. Gao et al., J. Biome.
d. Mater. Res. , Volume 55, 512-522
(2001, 2001), incompatible proteins often denature and lose their biological activity due to mixing processes.
【０００５】また、徐放キャリヤーとして親水性のハイ
ドロゲルを利用した例もあるが、タンパク質がハイドロ
ゲル内で自己拡散してしまうため、徐放の速度の制御が
難しい（Ｗ．Ｒ．Ｇｏｍｂｏｔｚ、Ｂｉｏｃｏｎｊｕｇ
ａｔｅ Ｃｈｅｍ．、６巻、３３２-３５１頁、１９９
５年）。There is also an example in which a hydrophilic hydrogel is used as a sustained release carrier, but since the protein self-diffuses in the hydrogel, it is difficult to control the sustained release rate (WR Gombottz, Bioconjug
ate Chem. , Vol. 6, pp. 332-351, 199
5 years).
【０００６】更に、生体分解性のハイドロゲルに成長因
子を固定し、生体内に埋め込むハイドロゲルは時間とと
もに分解し、成長因子の徐放が起こるといった系も報告
されている（Ｙ．Ｔａｂａｔａ等、Ｔｉｓｓｕｅ Ｅｎ
ｇｉｎｅｅｒｉｎｇ、５巻、１２７頁、１９９９年）。Further, a system has been reported in which a growth factor is immobilized on a biodegradable hydrogel, and the hydrogel embedded in the living body is decomposed with time, resulting in sustained release of the growth factor (Y. Tabata et al. Tissue En
Gineering, vol. 5, p. 127, 1999).
【０００７】しかし、いずれも非特異的なタンパク質の
包接を用いた手法であり、まだ改善の余地が残されてい
るばかりでなく、タンパク質と低分子の特異的な相互作
用をを用いた簡便な徐放化の手法は報告されていない。[0007] However, all of them are methods using non-specific inclusion of proteins, and there is still room for improvement. In addition, simple methods using specific interactions between proteins and small molecules are available. No sustained release method has been reported.
【０００８】[0008]
【発明が解決しようとする課題】本発明の目的は、疎水
性表面にコーティングすることにより、細胞培養等に使
用するタンパク質性の因子を、材料表面から徐放する機
能を付与する新規な構造を有するビオチン様ペプチド脂
質を提供することにある。SUMMARY OF THE INVENTION An object of the present invention is to provide a novel structure by coating a hydrophobic surface with a function of gradually releasing a proteinaceous factor used for cell culture or the like from the material surface. It is to provide a biotin-like peptide lipid having.
【０００９】また、ビオチン様ペプチド脂質を用いる、
培養素材等の疎水表面からのタンパク質徐放化法であ
る。Further, using a biotin-like peptide lipid,
This is a method for sustained release of proteins from the hydrophobic surface of culture materials and the like.
【００１０】[0010]
【課題を解決するための手段】Ｈｉｓ-Ｐｒｏ-Ｇｌｎ
（ＨＰＱ）配列のペプチドはアビジンとの弱い結合を持
つことが知られているが（Ｊ．Ｊ．Ｄｅｖｌｉｎ等、Ｓ
ｃｉｅｎｃｅ、２４９巻、４０４-４０６頁、１９９０
年）、このＨＰＱ配列をタンパク質等の徐放化に応用し
た例は知られていない。[Means for Solving the Problems] His-Pro-Gln
It is known that the peptide of the (HPQ) sequence has a weak bond with avidin (see J. Devlin et al., S.
science, 249, pp. 404-406, 1990
, Etc.), no example of applying this HPQ sequence to the sustained release of proteins and the like has been known.
【００１１】本発明者等は、ＨＰＱペプチドとアビジン
との弱い相互作用を利用すれば、タンパク質等の徐放化
のシステムを構築することが出来るのではないかと考
え、市販の安価な原料から短工程、高収率で合成でき、
コーティングするだけの簡単な操作で、培養素材からタ
ンパク質を徐放化する化合物として、式（１）で示され
るビオチン様ペプチド脂質を合成し、分子間相互作用解
析装置を用いて、ビオチン様ペプチド脂質が疎水性表面
にコーティングできること、さらにストレプトアビジン
を徐放化できることを確認して、本発明を完成するに至
った。The present inventors believe that a system for sustained-release of proteins and the like can be constructed by utilizing the weak interaction between HPQ peptide and avidin, and the short-term commercially available raw materials can Process, can be synthesized with high yield,
A biotin-like peptide lipid represented by the formula (1) was synthesized as a compound that slowly releases a protein from a culture material by a simple operation of coating, and the biotin-like peptide lipid was analyzed using an intermolecular interaction analyzer. The present invention was completed by confirming that the hydrophobic surface can be coated with and that streptavidin can be gradually released.
【化２】
（式中Ｒ1は、Ｏ（ＣＨ2）nＣＨ3基を、Ｒ2はＨまたは
Ｒ1を表し、ｌは０から３の整数を、ｍは２から６の整
数を、ｎは１１から１７の整数を、Ｈｉｓはヒスチジン
を、Ｐｒｏはプロリンを、Ｇｌｎはグルタミンを、Ｇｌ
ｙはグリシンを表わす。）[Chemical 2] (In the formula, R 1 represents an O (CH 2 ) n CH 3 group, R 2 represents H or R 1 , l is an integer of 0 to 3, m is an integer of 2 to 6, and n is 11 to An integer of 17, His for histidine, Pro for proline, Gln for glutamine and Gl.
y represents glycine. )
【００１２】本発明の化合物の合成はいかなる方法によ
ってもかまわない。３、４、５位に長鎖アルキロキシが
結合した安息香酸誘導体は、市販の化合物から文献既知
の方法（Ｖ．Ｓ．Ｋ．Ｂａｌａｇｕｒｕｓａｍｙ等、
Ｊ．Ａｍ．Ｃｈｅｍ．Ｓｏｃ．、１１９巻、１５３９-
１５５５頁、１９９７年）で合成できる。また、３、５
位に長鎖アルキロキシが結合した安息香酸誘導体も同様
に文献既知の方法（Ｖ．Ｐｅｒｃｅｃ等、Ａｎｇｅｗ．
Ｃｈｅｍ．Ｉｎｔ．Ｅｄ．、３９巻、１５９８-１６０
２頁、２０００年）で合成できる。The compound of the present invention may be synthesized by any method. A benzoic acid derivative having a long-chain alkyloxy group bonded at the 3, 4, and 5 positions can be obtained from commercially available compounds by a method known from the literature (VSK Balagurusami et al.
J. Am. Chem. Soc. 119, 1539-
1555, 1997). Also 3, 5
Similarly, a benzoic acid derivative having a long-chain alkyloxy group bonded at the position may be prepared by a method known in the literature (V. Percec et al., Angew.
Chem. Int. Ed. , 39, 1598-160
Page 2, 2000).
【００１３】その様にして得られた安息香酸誘導体を、
加水分解等により、遊離のカルボン酸とし、アルカンジ
アミンあるいはその一方のアミンを保護した化合物と、
例えば、通常のアミド結合形成に用いられる方法に従っ
て縮合させ必要に応じて脱保護することによって、アル
カンジアミンの片方のアミンと安息香酸がアミド結合し
た脂質を得ることが出来る。The benzoic acid derivative thus obtained is
A compound in which a free carboxylic acid is obtained by hydrolysis or the like and an alkanediamine or one of the amines is protected,
For example, a lipid in which one amine of alkanediamine and benzoic acid are amide-bonded can be obtained by condensation according to a method commonly used for amide bond formation and deprotection as necessary.
【００１４】式（１）で示されるペプチド脂質は、通常
のペプチド合成に用いられる方法に従って、アミノアル
キル基が結合した安息香酸誘導体に、順次、アミノ酸を
縮合することにより、あるいは、あらかじめ適当に保護
されたペプチド部分を合成し、それとアミノアルキル基
が結合した安息香酸誘導体を縮合することによって得ら
れ、必要に応じて、脱保護の後、シリカゲルカラム、ゲ
ルろ過等の方法によって精製を行うことによって得られ
る。The peptide lipid represented by the formula (1) can be protected by sequentially condensing amino acids with a benzoic acid derivative having an aminoalkyl group bonded thereto, or by appropriately preliminarily protecting it according to a method used for ordinary peptide synthesis. It is obtained by synthesizing the peptide portion thus prepared and condensing it with a benzoic acid derivative having an aminoalkyl group bonded, and if necessary, after deprotection, it is purified by a method such as a silica gel column or gel filtration. can get.
【００１５】タンパク質の徐放化は、ＨＰＱペプチド脂
質を、例えばエタノール等の適当な溶媒に溶解して培養
素材の表面にコートし、アビジン標識したタンパク質溶
液を用いるか、あるいは、一旦アビジンを結合させ、そ
の上にビオチン標識したタンパク質の水溶液を加えて、
徐放化したいタンパク質を素材表面に保持させる。その
上に、細胞培養液等を加えれば、保持されたタンパク質
が徐放される。アビジンまたはビオチンによる修飾は、
タンパク質の活性を保ったまま修飾する方法として確立
している（新生化学実験講座、１２巻、分子免疫学III
抗原・抗体・補体、９９頁-１０４頁、日本生化学会
編、東京化学同人、１９９２年）ので、この手法は、タ
ンパク質徐放化の一般的手法として用いることが出来
る。また、このタンパク質徐放化の手法は、従来から知
られている徐放キャリヤー等と併用することも出来る。For the sustained release of protein, HPQ peptide lipid is dissolved in a suitable solvent such as ethanol and coated on the surface of the culture material, and a protein solution labeled with avidin is used, or once avidin is bound. , Add an aqueous solution of biotin-labeled protein on it,
Hold the protein you want to release slowly on the material surface. If a cell culture solution or the like is added thereto, the retained protein will be released gradually. Modification with avidin or biotin
It has been established as a method for modifying proteins while maintaining their activity (Shinsei Chemistry Laboratory, Volume 12, Molecular Immunology III.
Antigen / antibody / complement, pp. 99-104, edited by The Biochemical Society of Japan, Tokyo Kagaku Dojin, 1992), so this method can be used as a general method for sustained release of proteins. Further, this method of sustained-release of protein can be used in combination with a conventionally known sustained-release carrier and the like.
【００１６】[0016]
【発明の実施の形態】本発明は式（１）で示されるペプ
チド脂質を提供するものであり、Ｈｉｓ-Ｐｒｏ-Ｇｌｎ
構造を有するタンパク質徐放素材を提供するものであ
り、式（１）で示されるペプチド脂質を有効成分とする
タンパク質徐放素材を提供するものであり、式（１）で
示されるペプチド脂質を有効成分とするタンパク質徐放
化剤を提供するものである。BEST MODE FOR CARRYING OUT THE INVENTION The present invention provides a peptide lipid represented by the formula (1), which is His-Pro-Gln.
The present invention provides a protein sustained-release material having a structure, which provides a protein sustained-release material containing a peptide lipid represented by the formula (1) as an active ingredient, and the peptide lipid represented by the formula (1) is effective. A protein sustained-release agent as a component is provided.
【００１７】以下に、本発明をさらに詳細に説明する
が、本発明は以下の記述に限定されるものではない。The present invention will be described in more detail below, but the present invention is not limited to the following description.
【００１８】[0018]
【実施例１】Ｎ-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-６-アミノ
ヘキシル）-３，５-ビス（ドデシロキシ）ベンズアミド
の合成
Ｆｍｏｃ-Ｇｌｎ（Ｔｒｔ）-ＯＨ (733 mg, 1.20 mmo
l)、水溶性カルボジイミド (230 mg, 1.20 mmol)、１-
ヒドロキシベンゾトリアゾール (184 mg, 1.20 mmol)を
乾燥ジクロロメタンに溶解し、室温で1時間撹拌した
後、Ｎ-（６-アミノヘキシル）-３，５-ビス（ドデシロ
キシ）ベンズアミド (588 mg, 1.00 mmol) のジクロロ
メタン溶液を加え、更に室温で２時間撹拌した。反応終
了後、反応溶液を水洗し、硫酸マグネシウムで乾燥させ
た。硫酸マグネシウムをろ別し、溶媒を減圧下留去し
た。残渣をＳｅｐｈａｄｅｘ ＬＨ-２０（ジクロロメ
タン：メタノール＝７：３）で精製し、Ｎ-（Ｎ-Ｇｌｎ
（Ｔｒｔ）（Ｆｍｏｃ）-６-アミノヘキシル）-３，５-
ビス（ドデシロキシ）ベンズアミド (1.09 g, 0.920 mm
ol,92%) を得た。Example 1 Synthesis of N- (N-His-Pro-Gln-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide Fmoc-Gln (Trt) -OH (733 mg, 1.20 mmo
l), water-soluble carbodiimide (230 mg, 1.20 mmol), 1-
Hydroxybenzotriazole (184 mg, 1.20 mmol) was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (588 mg, 1.00 mmol) Was added to the dichloromethane solution, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by Sephadex LH-20 (dichloromethane: methanol = 7: 3), N- (N-Gln
(Trt) (Fmoc) -6-aminohexyl) -3,5-
Bis (dodecyloxy) benzamide (1.09 g, 0.920 mm
ol, 92%).
【００１９】1H NMR (CDCl3, 40℃)δ0.89 (6H, t, J=
6.8 Hz), 1.20-1.58 (44H, m), 1.76(4H, quint, J=7.0
Hz), 1.92 (1H, m), 2.50 (1H, m), 2.40 (1H, brs),
2.55(1H, brs), 3.18 (2H, m), 3.35 (2H, quint, J=6.
2 Hz), 3.94 (4H, t, J=6.6Hz), 3.98 (1H, m), 4.19
(1H, t, J=7.1 Hz), 4.37 (2H, d, J=6.6 Hz), 5.94(1
H, brs), 6.11 (1H, brs), 6.31 (1H, brs), 6.54 (1H,
t, J=2.2 Hz), 6.86(2H, d, J=2.2 Hz), 6.98 (1H, br
s), 7.25 (17H, m), 7.38 (2H, t, J=7.6 Hz), 7.57 (2
H, d, J=7.6 Hz), 7.74 (2H, d, J=7.6 Hz).1H NMR (CDCl 3 , 40 ° C.) δ0.89 (6H, t, J =
6.8 Hz), 1.20-1.58 (44H, m), 1.76 (4H, quint, J = 7.0
Hz), 1.92 (1H, m), 2.50 (1H, m), 2.40 (1H, brs),
2.55 (1H, brs), 3.18 (2H, m), 3.35 (2H, quint, J = 6.
2 Hz), 3.94 (4H, t, J = 6.6Hz), 3.98 (1H, m), 4.19
(1H, t, J = 7.1 Hz), 4.37 (2H, d, J = 6.6 Hz), 5.94 (1
H, brs), 6.11 (1H, brs), 6.31 (1H, brs), 6.54 (1H,
t, J = 2.2 Hz), 6.86 (2H, d, J = 2.2 Hz), 6.98 (1H, br
s), 7.25 (17H, m), 7.38 (2H, t, J = 7.6 Hz), 7.57 (2
H, d, J = 7.6 Hz), 7.74 (2H, d, J = 7.6 Hz).
【００２０】Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）（Ｆｍｏｃ）-
６-アミノヘキシル）-３，５-ビス（ドデシロキシ）ベ
ンズアミド (600 mg, 0.508 mmol) をジクロロメタン
(2 ml) に溶解し、ピペリジン (1 ml) を加えて、2時間
撹拌した。反応溶液をＳｅｐｈａｄｅｘ ＬＨ-２０
（ジクロロメタン：メタノール＝７：３）で精製し、Ｎ
-（Ｎ−Ｇｌｎ（Ｔｒｔ）-６-アミノヘキシル）-３，５
-ビス（ドデシロキシ）ベンズアミド (472 mg, 0.492mm
ol, 97%) を得た。N- (N-Gln (Trt) (Fmoc)-
6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (600 mg, 0.508 mmol) was added to dichloromethane.
It was dissolved in (2 ml), piperidine (1 ml) was added, and the mixture was stirred for 2 hours. The reaction solution is Sephadex LH-20
Purify with (dichloromethane: methanol = 7: 3), N
-(N-Gln (Trt) -6-aminohexyl) -3,5
-Bis (dodecyloxy) benzamide (472 mg, 0.492mm
ol, 97%) was obtained.
【００２１】1H NMR (CDCl3)δ0.88 (6H, t, J=6.6 H
z), 1.25-1.55 (38H, m), 1.55-1.85(10H, m), 1.92 (2
H, m), 2.46 (2H, m), 3.06 (1H, dd, J=8.1, 5.1 Hz),
3.17(2H, t, J=6.8 Hz), 3.35 (2H, m), 3.96 (4H, t,
J=6.4 Hz), 6.56 (1H, t, J=1.7 H), 6.82 (2H, d, J=
1.7 Hz), 7.25 (15H, m).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.6 H
z), 1.25-1.55 (38H, m), 1.55-1.85 (10H, m), 1.92 (2
H, m), 2.46 (2H, m), 3.06 (1H, dd, J = 8.1, 5.1 Hz),
3.17 (2H, t, J = 6.8 Hz), 3.35 (2H, m), 3.96 (4H, t,
J = 6.4 Hz), 6.56 (1H, t, J = 1.7 H), 6.82 (2H, d, J =
1.7 Hz), 7.25 (15H, m).
【００２２】Ｆｍｏｃ-Ｐｒｏ-ＯＨ (200 mg, 0.590 mm
ol)、水溶性カルボジイミド (113 mg,0.590 mmol)、１-
ヒドロキシベンゾトリアゾール (90 mg, 0.590 mmol)
を乾燥ジクロロメタンに溶解し、室温で1時間撹拌した
後、Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (470 mg,
0.490 mmol)のジクロロメタン溶液を加え、更に室温で
２時間撹拌した。反応終了後、反応溶液を水洗し、硫酸
マグネシウムで乾燥させた。硫酸マグネシウムをろ別
し、溶媒を減圧下留去した。残渣をＳｅｐｈａｄｅｘ
ＬＨ-２０（ジクロロメタン：メタノール＝７：３）で
精製し、Ｎ-（Ｎ-Ｐｒｏ（Ｆｍｏｃ）-Ｇｌｎ（Ｔｒ
ｔ）-６-アミノヘキシル）-３，５-ビス（ドデシロキ
シ）ベンズアミド (578 mg, 0.452 mmol, 92%) を得
た。Fmoc-Pro-OH (200 mg, 0.590 mm
ol), water-soluble carbodiimide (113 mg, 0.590 mmol), 1-
Hydroxybenzotriazole (90 mg, 0.590 mmol)
Was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Gln (Trt) -6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (470 mg,
A dichloromethane solution of 0.490 mmol) was added, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. Sephadex the residue
Purified with LH-20 (dichloromethane: methanol = 7: 3), N- (N-Pro (Fmoc) -Gln (Tr
t) -6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (578 mg, 0.452 mmol, 92%) was obtained.
【００２３】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.20-1.55 (44H, m), 1.75 (4H,m), 1.96 (2H, br
s), 2.11 (2H, brs), 2.43 (1H, brs), 2.53 (1H, ddd,
J=15.4, 8.6, 3.9 Hz), 3.15-3.30 (6H, m), 3.94 (4
H, t, J=6.6 Hz), 4.13 (3H, m), 4.35 (2H, m), 6.28
(1H, brs), 6.53 (1H, t, J=2.2 Hz), 6.85 (2H, d, J=
2.2 Hz), 6.88 (1H, brs), 6.94 (1H, brs), 7.20 (17
H, m), 7.38 (2H, m), 7.50 (2H, t, J=7.6 Hz), 7.75
(2H, d, J=7.6 Hz), 8.11 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.20-1.55 (44H, m), 1.75 (4H, m), 1.96 (2H, br
s), 2.11 (2H, brs), 2.43 (1H, brs), 2.53 (1H, ddd,
J = 15.4, 8.6, 3.9 Hz), 3.15-3.30 (6H, m), 3.94 (4
H, t, J = 6.6 Hz), 4.13 (3H, m), 4.35 (2H, m), 6.28
(1H, brs), 6.53 (1H, t, J = 2.2 Hz), 6.85 (2H, d, J =
2.2 Hz), 6.88 (1H, brs), 6.94 (1H, brs), 7.20 (17
H, m), 7.38 (2H, m), 7.50 (2H, t, J = 7.6 Hz), 7.75
(2H, d, J = 7.6 Hz), 8.11 (1H, brs).
【００２４】Ｎ-（Ｎ-Ｐｒｏ（Ｆｍｏｃ）-Ｇｌｎ（Ｔ
ｒｔ）-６-アミノヘキシル）-３，５-ビス（ドデシロキ
シ）ベンズアミド (578 mg, 0.452 mmol) をジクロロメ
タン (2 ml) に溶解し、ピペリジン (1 ml) を加えて、
2時間撹拌した。反応溶液をＳｅｐｈａｄｅｘ ＬＨ-２
０（ジクロロメタン：メタノール＝７：３）で精製し、
Ｎ-（Ｎ-Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-６-アミノヘキシ
ル）-３，５-ビス（ドデシロキシ）ベンズアミド (445
mg, 0.422 mmol, 93%) を得た。N- (N-Pro (Fmoc) -Gln (T
rt) -6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (578 mg, 0.452 mmol) is dissolved in dichloromethane (2 ml), piperidine (1 ml) is added,
It was stirred for 2 hours. The reaction solution is Sephadex LH-2
0 (dichloromethane: methanol = 7: 3),
N- (N-Pro-Gln (Trt) -6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (445
mg, 0.422 mmol, 93%) was obtained.
【００２５】1H NMR (CDCl3, 40℃)δ0.88 (6H, t, J=
6.8 Hz), 1.22-1.46 (42H, m), 1.55(2H, quint, J=6.8
Hz), 1.67 (2H, quint, J=6.8 Hz), 1.76 (4H, quint,
J=6.8 Hz), 1.84 (2H, m), 2.07 (2H, m), 2.35 (1H,
dt, J=14.9, 6.4 Hz), 2.52(1H, ddd, J=14.9, 8.5, 5.
9 Hz), 2.87 (1H, dt, J=10.1, 6.4 Hz), 2.96 (1H,dt,
J=10.1, 6.8 Hz), 3.16 (2H, q, J=6.4 Hz), 3.34 (2
H, dd, J=7.1, 2.9 Hz), 3.37 (2H, dd, J=6.8, 2.9 H
z), 3.66 (1H, dd, J=9.3, 5.6 Hz), 3.95 (4H, t, J=
6.6 Hz), 4.11 (1H, td, J=7.8, 5.4 Hz), 6.28 (1H,
t, J=5.8 Hz), 6.38 (1H, t, J=5.8 Hz), 6.53 (1H, t,
J=2.2 Hz), 6.87 (2H, d, J=2.2 Hz), 7.25 (15H, m),
8.21 (1H, d, J=7.8 Hz).1H NMR (CDCl 3 , 40 ° C.) δ 0.88 (6H, t, J =
6.8 Hz), 1.22-1.46 (42H, m), 1.55 (2H, quint, J = 6.8
Hz), 1.67 (2H, quint, J = 6.8 Hz), 1.76 (4H, quint,
J = 6.8 Hz), 1.84 (2H, m), 2.07 (2H, m), 2.35 (1H,
dt, J = 14.9, 6.4 Hz), 2.52 (1H, ddd, J = 14.9, 8.5, 5.
9 Hz), 2.87 (1H, dt, J = 10.1, 6.4 Hz), 2.96 (1H, dt,
J = 10.1, 6.8 Hz), 3.16 (2H, q, J = 6.4 Hz), 3.34 (2
H, dd, J = 7.1, 2.9 Hz), 3.37 (2H, dd, J = 6.8, 2.9 H
z), 3.66 (1H, dd, J = 9.3, 5.6 Hz), 3.95 (4H, t, J =
6.6 Hz), 4.11 (1H, td, J = 7.8, 5.4 Hz), 6.28 (1H,
t, J = 5.8 Hz), 6.38 (1H, t, J = 5.8 Hz), 6.53 (1H, t,
J = 2.2 Hz), 6.87 (2H, d, J = 2.2 Hz), 7.25 (15H, m),
8.21 (1H, d, J = 7.8 Hz).
【００２６】Ｆｍｏｃ-Ｈｉｓ（Ｔｒｔ）-ＯＨ (314 m
g, 0.506 mmol)、水溶性カルボジイミド (97 mg, 0.506
mmol)、１-ヒドロキシベンゾトリアゾール (77 mg, 0.
506mmol) を乾燥ジクロロメタンに溶解し、室温で1時間
撹拌した後、Ｎ-（Ｎ-Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-６-ア
ミノヘキシル）-３，５-ビス（ドデシロキシ）ベンズア
ミド (445 mg, 0.422 mmol) のジクロロメタン溶液を加
え、更に室温で２時間撹拌した。反応終了後、反応溶液
を水洗し、硫酸マグネシウムで乾燥させた。硫酸マグネ
シウムをろ別し、溶媒を減圧下留去した。残渣をＳｅｐ
ｈａｄｅｘＬＨ-２０（ジクロロメタン：メタノール＝
７：３）で精製し、Ｎ-（Ｎ-Ｈｉｓ（Ｔｒｔ）（Ｆｍｏ
ｃ）-Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (660 mg,
0.398 mmol, 95%) を得た。Fmoc-His (Trt) -OH (314 m
g, 0.506 mmol), water-soluble carbodiimide (97 mg, 0.506
mmol), 1-hydroxybenzotriazole (77 mg, 0.1.
506 mmol) was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Pro-Gln (Trt) -6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (445 mg, 0.422 mmol). A dichloromethane solution of) was added, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue is Sep
hadexLH-20 (dichloromethane: methanol =
7: 3) and purified with N- (N-His (Trt) (Fmo
c) -Pro-Gln (Trt) -6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (660 mg,
0.398 mmol, 95%) was obtained.
【００２７】1H NMR (CDCl3)δ0.88 (4H, t, J=5.8 H),
1.23-1.48 (H, m), 1.55 (3H, m),1.74 (4H, quint, J
=6.8 Hz), 1.82 (1H, m), 1.90 (2H, m), 2.35 (1H, qu
int, J=7.3 Hz), 2.40 (1H, quint, J=7.3 Hz), 2.82-
3.08 (4H, m), 3.28 (1H, m), 3.36 (2H, q, J=6.6 H
z), 3.54 (1H, brs), 3.93 (4H, t, J=6.6 Hz), 4.14
(1H, t, J=6.8 Hz), 4.33 (3H, m), 4.51 (1H, brs),
6.06 (1H, d, J=7.8 Hz),6.53 (1H, t, J=2.2 Hz), 6.7
0 (1H, s), 6.88 (2H, d, J=2.2 Hz), 6.93 (1H,brs),
7.06-7.32 (32H, m), 7.37 (2H, t, J=7.6 Hz), 7.45
(1H, s), 7.54 (2H, d, J=7.6 Hz), 7.73 (2H, d, J=7.
6 Hz), 9.07 (1H, d, J=7.8 Hz).1H NMR (CDCl 3 ) δ 0.88 (4H, t, J = 5.8H),
1.23-1.48 (H, m), 1.55 (3H, m), 1.74 (4H, quint, J
= 6.8 Hz), 1.82 (1H, m), 1.90 (2H, m), 2.35 (1H, qu
int, J = 7.3 Hz), 2.40 (1H, quint, J = 7.3 Hz), 2.82-
3.08 (4H, m), 3.28 (1H, m), 3.36 (2H, q, J = 6.6 H
z), 3.54 (1H, brs), 3.93 (4H, t, J = 6.6 Hz), 4.14
(1H, t, J = 6.8 Hz), 4.33 (3H, m), 4.51 (1H, brs),
6.06 (1H, d, J = 7.8 Hz), 6.53 (1H, t, J = 2.2 Hz), 6.7
0 (1H, s), 6.88 (2H, d, J = 2.2 Hz), 6.93 (1H, brs),
7.06-7.32 (32H, m), 7.37 (2H, t, J = 7.6 Hz), 7.45
(1H, s), 7.54 (2H, d, J = 7.6 Hz), 7.73 (2H, d, J = 7.
6 Hz), 9.07 (1H, d, J = 7.8 Hz).
【００２８】Ｎ-（Ｎ-Ｈｉｓ（Ｔｒｔ）（Ｆｍｏｃ）-
Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-６-アミノヘキシル）-３，５
-ビス（ドデシロキシ）ベンズアミド (100 mg, 0.060 m
mol)にトリフルオロ酢酸 (0.5 ml) を加えて、2時間撹
拌した。トリフルオロ酢酸を減圧下留去し、シリカゲル
カラムクロマトグラフィー（クロロホルム：メタノール
＝９５：５）で精製し、Ｎ-（Ｎ-Ｈｉｓ（Ｆｍｏｃ）-
Ｐｒｏ-Ｇｌｎ-６-アミノヘキシル）-３，５-ビス（ド
デシロキシ）ベンズアミド (68 mg, 0.058 mmol,97%)
を得た。N- (N-His (Trt) (Fmoc)-
Pro-Gln (Trt) -6-aminohexyl) -3,5
-Bis (dodecyloxy) benzamide (100 mg, 0.060 m
Trifluoroacetic acid (0.5 ml) was added to (mol) and the mixture was stirred for 2 hours. Trifluoroacetic acid was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 95: 5) and N- (N-His (Fmoc)-
Pro-Gln-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (68 mg, 0.058 mmol, 97%)
Got
【００２９】1H NMR (CDCl3)δ0.88 (3H, t, J=6.8 H
z), 1.20-1.50 (40H, m), 1.54 (4H,m), 1.76 (4H, m),
1.90 (2H, m), 2.01 (2H, m), 2.38 (3H, m), 2.40 (1
H, m), 3.01 (2H, m), 3.22 (2H, m), 3.35 (2H, dd, J
=13.7, 6.6 Hz), 3.55 (1H, dd, J=16.1, 6.6 Hz), 3.9
5 (4H, t, J=6.6 Hz), 4.20 (1H, t, J=6.8 Hz), 4.42
(3H, m), 4.58 (1H, dd, J=8.9, 4.1 Hz), 6.55 (1H,
s), 6.88 (2H, s), 6.92(1H, brs), 7.04 (1H, t, J=5.
6 Hz), 7.30 (2H, t, J=7.6 Hz), 7.37 (2H, t,J=7.6 H
z), 7.55 (1H, s), 7.57 (2H, d, J=7.6 Hz), 7.74 (1
H, s), 7.76 (2H, d, J=7.6Hz).1 H NMR (CDCl 3 ) δ 0.88 (3H, t, J = 6.8 H
z), 1.20-1.50 (40H, m), 1.54 (4H, m), 1.76 (4H, m),
1.90 (2H, m), 2.01 (2H, m), 2.38 (3H, m), 2.40 (1
H, m), 3.01 (2H, m), 3.22 (2H, m), 3.35 (2H, dd, J
= 13.7, 6.6 Hz), 3.55 (1H, dd, J = 16.1, 6.6 Hz), 3.9
5 (4H, t, J = 6.6 Hz), 4.20 (1H, t, J = 6.8 Hz), 4.42
(3H, m), 4.58 (1H, dd, J = 8.9, 4.1 Hz), 6.55 (1H,
s), 6.88 (2H, s), 6.92 (1H, brs), 7.04 (1H, t, J = 5.
6 Hz), 7.30 (2H, t, J = 7.6 Hz), 7.37 (2H, t, J = 7.6 H
z), 7.55 (1H, s), 7.57 (2H, d, J = 7.6 Hz), 7.74 (1
H, s), 7.76 (2H, d, J = 7.6Hz).
【００３０】Ｎ-（Ｎ-Ｈｉｓ（Ｆｍｏｃ）-Ｐｒｏ-Ｇｌ
ｎ-６-アミノヘキシル）-３，５-ビス（ドデシロキシ）
ベンズアミド (30 mg, 0.026 mmol) をジクロロメタン
(2 ml) に溶解し、ピペリジン (1 ml) を加えて、2時間
撹拌した。反応溶液をＳｅｐｈａｄｅｘ ＬＨ-２０
（ジクロロメタン：メタノール＝７：３）で精製し、Ｎ
-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (19mg, 0.0
20mmol, 75%) を得た。N- (N-His (Fmoc) -Pro-Gl
n-6-aminohexyl) -3,5-bis (dodecyloxy)
Benzamide (30 mg, 0.026 mmol) in dichloromethane
It was dissolved in (2 ml), piperidine (1 ml) was added, and the mixture was stirred for 2 hours. The reaction solution is Sephadex LH-20
Purify with (dichloromethane: methanol = 7: 3), N
-(N-His-Pro-Gln-6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (19mg, 0.0
20 mmol, 75%) was obtained.
【００３１】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.23-1.62 (44H, m), 1.77 (4H,quint, J=6.6 Hz),
1.95 (4H, m), 2.23 (3H, m), 2.39 (1H, quint, J=7.
6 Hz), 3.09-3.30 (4H, m), 3.35 (2H, t, J=7.3 Hz),
3.38 (2H, quint, J=1.7 Hz),3.60 (1H, m), 3.98 (4H,
t, J=6.6 Hz), 4.12 (1H, t, J=6.3 Hz), 4.42 (2H,
m), 6.56 (1H, t, J=2.2 Hz), 6.88 (2H, d, J=2.2 H
z), 7.33 (1H, d, J=1.2Hz), 7.88 (1H, d, J=1.2 Hz).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.23-1.62 (44H, m), 1.77 (4H, quint, J = 6.6 Hz),
1.95 (4H, m), 2.23 (3H, m), 2.39 (1H, quint, J = 7.
6 Hz), 3.09-3.30 (4H, m), 3.35 (2H, t, J = 7.3 Hz),
3.38 (2H, quint, J = 1.7 Hz), 3.60 (1H, m), 3.98 (4H,
t, J = 6.6 Hz), 4.12 (1H, t, J = 6.3 Hz), 4.42 (2H,
m), 6.56 (1H, t, J = 2.2 Hz), 6.88 (2H, d, J = 2.2 H
z), 7.33 (1H, d, J = 1.2Hz), 7.88 (1H, d, J = 1.2 Hz).
【００３２】[0032]
【実施例２】Ｎ-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-Ｇｌｙ-Ｇ
ｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス（ドデ
シロキシ）ベンズアミドの合成
Ｆｍｏｃ-Ｇｌｙ-ＯＨ (356 mg, 1.20 mmol)、水溶性カ
ルボジイミド (230 mg, 1.20 mmol)、１-ヒドロキシベ
ンゾトリアゾール (184 mg, 1.20 mmol) を乾燥ジクロ
ロメタンに溶解し、室温で1時間撹拌した後、Ｎ-（６-
アミノヘキシル）-３，５-ビス（ドデシロキシ）ベンズ
アミド (588 mg, 1.00 mmol) のジクロロメタン溶液を
加え、更に室温で２時間撹拌した。反応終了後、反応溶
液を水洗し、硫酸マグネシウムで乾燥させた。硫酸マグ
ネシウムをろ別し、溶媒を減圧下留去した。残渣をシリ
カゲルカラムクロマトグラフィー（クロロホルム：メタ
ノール＝９８：２）で精製し、Ｎ-（Ｎ-Ｇｌｙ（Ｆｍｏ
ｃ）-６-アミノヘキシル）-３，５-ビス（ドデシロキ
シ）ベンズアミド (798 mg, 0.920 mmol, 92%) を得
た。Example 2 N- (N-His-Pro-Gln-Gly-G
Synthesis of ly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide Fmoc-Gly-OH (356 mg, 1.20 mmol), water-soluble carbodiimide (230 mg, 1.20 mmol), 1-hydroxybenzotriazole (184 mg, 1.20 mmol) was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (6-
A solution of aminohexyl) -3,5-bis (dodecyloxy) benzamide (588 mg, 1.00 mmol) in dichloromethane was added, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 98: 2), and N- (N-Gly (Fmo
c) -6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (798 mg, 0.920 mmol, 92%) was obtained.
【００３３】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.25-1.60 (44H, m), 1.75 (4H,q, J=6.8 Hz), 3.2
8 (2H, q, J=6.3 Hz), 3.42 (2H, q, J=6.3 Hz), 3.86
(2H,d, J=6.3 Hz), 3.94 (4H, t, J=6.6 Hz), 5.71 (1
H, brs), 6.14 (1H, t, J=5.3Hz), 6.18 (1H, brs), 6.
54 (1H, t, J=2.2 Hz), 6.85 (2H, d, J=2.2 Hz), 7.29
(2H, t, J=7.6 Hz), 7.37 (2H, t, J=7.6 Hz), 7.58
(2H, d, J=7.6 Hz), 7.75 (2H, d, J=7.6 Hz).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.25-1.60 (44H, m), 1.75 (4H, q, J = 6.8 Hz), 3.2
8 (2H, q, J = 6.3 Hz), 3.42 (2H, q, J = 6.3 Hz), 3.86
(2H, d, J = 6.3 Hz), 3.94 (4H, t, J = 6.6 Hz), 5.71 (1
H, brs), 6.14 (1H, t, J = 5.3Hz), 6.18 (1H, brs), 6.
54 (1H, t, J = 2.2 Hz), 6.85 (2H, d, J = 2.2 Hz), 7.29
(2H, t, J = 7.6 Hz), 7.37 (2H, t, J = 7.6 Hz), 7.58
(2H, d, J = 7.6 Hz), 7.75 (2H, d, J = 7.6 Hz).
【００３４】Ｎ-（Ｎ-Ｇｌｙ（Ｆｍｏｃ）-６-アミノヘ
キシル）-３，５-ビス（ドデシロキシ）ベンズアミド
(737 mg, 0.849 mmol) をジクロロメタン (2 ml) に溶
解し、ピペリジン (1 ml) を加えて、2時間撹拌した。
反応溶液をＳｅｐｈａｄｅｘＬＨ-２０（ジクロロメタ
ン：メタノール＝７：３）で精製し、Ｎ-（Ｎ-Ｇｌｙ-
６-アミノヘキシル）-３，５-ビス（ドデシロキシ）ベ
ンズアミド (522 mg, 0.809 mmol, 95%) を得た。N- (N-Gly (Fmoc) -6-aminohexyl) -3,5-bis (dodecyloxy) benzamide
(737 mg, 0.849 mmol) was dissolved in dichloromethane (2 ml), piperidine (1 ml) was added, and the mixture was stirred for 2 hours.
The reaction solution was purified with Sephadex LH-20 (dichloromethane: methanol = 7: 3), and N- (N-Gly-
6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (522 mg, 0.809 mmol, 95%) was obtained.
【００３５】1H NMR (CDCl3)δ0.90 (6H, t, J=6.8 H
z), 1.25-1.60 (44H, m), 1.77 (4H,quint, J=6.8 Hz),
3.29 (2H, q, J=6.6 Hz), 3.42 (2H, q, J=6.8 Hz),
3.94 (2H, m), 3.96 (4H, t, J=6.6 Hz), 6.21 (1H, t,
J=5.6 Hz), 6.55 (1H, t, J=2.2 Hz), 6.86 (2H, d, J
=2.2 Hz), 7.29 (1H, brs).1 H NMR (CDCl 3 ) δ 0.90 (6H, t, J = 6.8 H
z), 1.25-1.60 (44H, m), 1.77 (4H, quint, J = 6.8 Hz),
3.29 (2H, q, J = 6.6 Hz), 3.42 (2H, q, J = 6.8 Hz),
3.94 (2H, m), 3.96 (4H, t, J = 6.6 Hz), 6.21 (1H, t,
J = 5.6 Hz), 6.55 (1H, t, J = 2.2 Hz), 6.86 (2H, d, J
= 2.2 Hz), 7.29 (1H, brs).
【００３６】Ｆｍｏｃ-Ｇｌｙ-ＯＨ (276 mg, 0.930 mm
ol)、水溶性カルボジイミド (178 mg, 0.930 mmol)、１
-ヒドロキシベンゾトリアゾール (142 mg, 0.930 mmol)
を乾燥ジクロロメタンに溶解し、室温で1時間撹拌した
後、Ｎ-（Ｎ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス
（ドデシロキシ）ベンズアミド (500 mg, 0.775 mmol)
のジクロロメタン溶液を加え、更に室温で２時間撹拌し
た。反応終了後、反応溶液を水洗し、硫酸マグネシウム
で乾燥させた。硫酸マグネシウムをろ別し、溶媒を減圧
下留去した。残渣をＳｅｐｈａｄｅｘ ＬＨ-２０（ジ
クロロメタン：メタノール＝７：３）で精製し、Ｎ-
（Ｎ-Ｇｌｙ（Ｆｍｏｃ）-Ｇｌｙ-６-アミノヘキシル）
-３，５-ビス（ドデシロキシ）ベンズアミド (674 mg,
0.729 mmol, 94%) を得た。Fmoc-Gly-OH (276 mg, 0.930 mm
ol), water-soluble carbodiimide (178 mg, 0.930 mmol), 1
-Hydroxybenzotriazole (142 mg, 0.930 mmol)
Was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (500 mg, 0.775 mmol)
Was added to the dichloromethane solution, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by Sephadex LH-20 (dichloromethane: methanol = 7: 3) and N-
(N-Gly (Fmoc) -Gly-6-aminohexyl)
-3,5-bis (dodecyloxy) benzamide (674 mg,
0.729 mmol, 94%) was obtained.
【００３７】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.25-1.68 (44H, m), 1.74 (4H,quint, J=6.6 Hz),
3.24 (2H, q, J=6.6 Hz), 3.40 (2H, q, J=6.6 Hz),
3.87 (2H, m), 3.92 (2H, m), 3.93 (4H, t, J=6.6 H
z), 4.19 (1H, t, J=6.8 Hz), 4.43 (2H, d, J=6.8 H
z), 5.83 (1H, brs), 6.31 (1H, brs), 6.54 (1H, t, J
=2.2Hz), 6.85 (2H, d, J=2.2 Hz), 6.98 (1H, brs),
7.30 (2H, t, J=7.6 Hz), 7.39 (2H, t, J=7.6 Hz), 7.
55 (2H, d, J=7.6 Hz), 7.75 (2H, d, J=7.6 Hz).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.25-1.68 (44H, m), 1.74 (4H, quint, J = 6.6 Hz),
3.24 (2H, q, J = 6.6 Hz), 3.40 (2H, q, J = 6.6 Hz),
3.87 (2H, m), 3.92 (2H, m), 3.93 (4H, t, J = 6.6 H
z), 4.19 (1H, t, J = 6.8 Hz), 4.43 (2H, d, J = 6.8 H
z), 5.83 (1H, brs), 6.31 (1H, brs), 6.54 (1H, t, J
= 2.2Hz), 6.85 (2H, d, J = 2.2 Hz), 6.98 (1H, brs),
7.30 (2H, t, J = 7.6 Hz), 7.39 (2H, t, J = 7.6 Hz), 7.
55 (2H, d, J = 7.6 Hz), 7.75 (2H, d, J = 7.6 Hz).
【００３８】Ｎ-（Ｎ-Ｇｌｙ（Ｆｍｏｃ）-Ｇｌｙ-６-
アミノヘキシル）-３，５-ビス（ドデシロキシ）ベンズ
アミド (600 mg, 0.649 mmol) をジクロロメタン (2 m
l) に溶解し、ピペリジン (1 ml) を加えて、2時間撹拌
した。反応溶液をＳｅｐｈａｄｅｘ ＬＨ-２０（ジク
ロロメタン：メタノール＝７：３）で精製し、Ｎ-（Ｎ-
Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス（ド
デシロキシ）ベンズアミド (438 mg, 0.623 mmol, 96%)
を得た。N- (N-Gly (Fmoc) -Gly-6-
Aminohexyl) -3,5-bis (dodecyloxy) benzamide (600 mg, 0.649 mmol) was added to dichloromethane (2 m
It was dissolved in l), piperidine (1 ml) was added, and the mixture was stirred for 2 hours. The reaction solution was purified with Sephadex LH-20 (dichloromethane: methanol = 7: 3), and N- (N-
Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (438 mg, 0.623 mmol, 96%)
Got
【００３９】1H NMR (CDCl3)δ0.88 (6H, t, J=6.6 H
z), 1.25-1.59 (42H, m), 1.75 (4H,q, J=6.6 Hz), 2.1
5 (2H, brs), 3.23 (2H, q, J=6.3 Hz), 3.39 (4H, m),
3.94(4H, t, J=6.3 Hz), 4.00 (2H, m), 6.54 (1H,
s), 6.62 (1H, t, J=5.1 Hz),6.86 (1H, brs), 6.89 (2
H, s), 8.00 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.6 H
z), 1.25-1.59 (42H, m), 1.75 (4H, q, J = 6.6 Hz), 2.1
5 (2H, brs), 3.23 (2H, q, J = 6.3 Hz), 3.39 (4H, m),
3.94 (4H, t, J = 6.3 Hz), 4.00 (2H, m), 6.54 (1H,
s), 6.62 (1H, t, J = 5.1 Hz), 6.86 (1H, brs), 6.89 (2
H, s), 8.00 (1H, brs).
【００４０】Ｆｍｏｃ-Ｇｌｙ-ＯＨ (222 mg, 0.747 mm
ol)、水溶性カルボジイミド (143 mg, 0.747 mmol)、１
-ヒドロキシベンゾトリアゾール (114 mg, 0.747 mmol)
を乾燥ジクロロメタンに溶解し、室温で1時間撹拌した
後、Ｎ-（Ｎ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，
５-ビス（ドデシロキシ）ベンズアミド (350 mg, 0.498
mmol) のジクロロメタン溶液を加え、更に室温で２時
間撹拌した。反応終了後、反応溶液を水洗し、硫酸マグ
ネシウムで乾燥させた。硫酸マグネシウムをろ別し、溶
媒を減圧下留去した。残渣をＳｅｐｈａｄｅｘ ＬＨ-
２０（ジクロロメタン：メタノール＝７：３）で精製
し、Ｎ-（Ｎ-Ｇｌｙ（Ｆｍｏｃ）-Ｇｌｙ-Ｇｌｙ-６-ア
ミノヘキシル）-３，５-ビス（ドデシロキシ）ベンズア
ミド (478mg, 0.487 mmol, 98%) を得た。Fmoc-Gly-OH (222 mg, 0.747 mm
ol), water-soluble carbodiimide (143 mg, 0.747 mmol), 1
-Hydroxybenzotriazole (114 mg, 0.747 mmol)
Was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Gly-Gly-6-aminohexyl) -3,3
5-bis (dodecyloxy) benzamide (350 mg, 0.498
Dichloromethane solution) was added, and the mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue is Sephadex LH-
Purify with 20 (dichloromethane: methanol = 7: 3) and N- (N-Gly (Fmoc) -Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (478 mg, 0.487 mmol, 98). %)
【００４１】1H NMR (CDCl3)δ0.88 (6H, t, J=6.6 H
z), 1.25-1.53 (42H, m), 1.59 (2H,m), 1.75 (4H, q,
J=7.0 Hz), 3.24 (2H, q, J=6.3 Hz), 3.40 (2H, q, J=
6.8 Hz), 3.86 (6H, m), 3.94 (4H, t, J=6.6 Hz), 4.1
9 (1H, t, J=6.8 Hz), 4.42 (2H, d, J=6.8 Hz), 5.82
(1H, brs), 6.25 (1H, t, J=5.4 Hz), 6.44 (1H, brs),
6.55 (1H, t, J=2.2 Hz), 6.83 (2H, d, J=2.2 Hz),
6.92 (1H, brs), 7.17 (1H, brs), 7.22 (1H, brs), 7.
28 (2H, t, J=7.3 Hz), 7.36 (2H, t, J=7.3 Hz), 7.56
(2H, d, J=7.3 Hz), 7.74 (2H, d, J=7.3 Hz).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.6 H
z), 1.25-1.53 (42H, m), 1.59 (2H, m), 1.75 (4H, q,
J = 7.0 Hz), 3.24 (2H, q, J = 6.3 Hz), 3.40 (2H, q, J =
6.8 Hz), 3.86 (6H, m), 3.94 (4H, t, J = 6.6 Hz), 4.1
9 (1H, t, J = 6.8 Hz), 4.42 (2H, d, J = 6.8 Hz), 5.82
(1H, brs), 6.25 (1H, t, J = 5.4 Hz), 6.44 (1H, brs),
6.55 (1H, t, J = 2.2 Hz), 6.83 (2H, d, J = 2.2 Hz),
6.92 (1H, brs), 7.17 (1H, brs), 7.22 (1H, brs), 7.
28 (2H, t, J = 7.3 Hz), 7.36 (2H, t, J = 7.3 Hz), 7.56
(2H, d, J = 7.3 Hz), 7.74 (2H, d, J = 7.3 Hz).
【００４２】Ｎ-（Ｎ-Ｇｌｙ（Ｆｍｏｃ）-Ｇｌｙ-Ｇｌ
ｙ-６-アミノヘキシル）-３，５-ビス（ドデシロキシ）
ベンズアミド (300 mg, 0.306 mmol) をジクロロメタン
(2ml) に溶解し、ピペリジン (1 ml) を加えて、2時間
撹拌した。反応溶液をＳｅｐｈａｄｅｘ ＬＨ-２０
（ジクロロメタン：メタノール＝７：３）で精製し、Ｎ
-（Ｎ-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (220 mg,
0.290 mmol, 95%) を得た。N- (N-Gly (Fmoc) -Gly-Gl
y-6-aminohexyl) -3,5-bis (dodecyloxy)
Benzamide (300 mg, 0.306 mmol) in dichloromethane
It was dissolved in (2 ml), piperidine (1 ml) was added, and the mixture was stirred for 2 hours. The reaction solution is Sephadex LH-20
Purify with (dichloromethane: methanol = 7: 3), N
-(N-Gly-Gly-Gly-6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (220 mg,
0.290 mmol, 95%) was obtained.
【００４３】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.23-1.50 (40H, m), 1.52 (2H,m), 1.60 (2H, m),
1.70 (4H, m), 3.26 (2H, q, J=6.3 Hz), 3.43 (4H,
m), 3.94 (8H, m), 6.43 (1H, brs), 6.55 (1H, t, J=
2.0 Hz), 6.64 (1H, brs), 6.86(2H, d, J=2.0 Hz), 7.
17 (1H, brs), 8.01 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.23-1.50 (40H, m), 1.52 (2H, m), 1.60 (2H, m),
1.70 (4H, m), 3.26 (2H, q, J = 6.3 Hz), 3.43 (4H,
m), 3.94 (8H, m), 6.43 (1H, brs), 6.55 (1H, t, J =
2.0 Hz), 6.64 (1H, brs), 6.86 (2H, d, J = 2.0 Hz), 7.
17 (1H, brs), 8.01 (1H, brs).
【００４４】Ｆｍｏｃ-Ｇｌｎ（Ｔｒｔ）-ＯＨ (362mg,
0.592 mmol)、水溶性カルボジイミド (114 mg, 0.592
mmol)、１-ヒドロキシベンゾトリアゾール (91 mg, 0.5
92 mmol) を乾燥ジクロロメタンに溶解し、室温で1時間
撹拌した後、Ｎ-（Ｎ-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノ
ヘキシル）-３，５-ビス（ドデシロキシ）ベンズアミド
(300 mg, 0.395 mmol) のジクロロメタン溶液を加え、
更に室温で２時間撹拌した。反応終了後、反応溶液を水
洗し、硫酸マグネシウムで乾燥させた。硫酸マグネシウ
ムをろ別し、溶媒を減圧下留去した。残渣をシリカゲル
カラムクロマトグラフィー（クロロホルム：メタノール
＝９８：２）で精製し、Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）（Ｆ
ｍｏｃ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (482 mg,
0.357 mmol, 90%) を得た。Fmoc-Gln (Trt) -OH (362 mg,
0.592 mmol), water-soluble carbodiimide (114 mg, 0.592
mmol), 1-hydroxybenzotriazole (91 mg, 0.5
(92 mmol) was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Gly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide
Add a solution of (300 mg, 0.395 mmol) in dichloromethane,
The mixture was further stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 98: 2), and N- (N-Gln (Trt) (F
moc) -Gly-Gly-Gly-6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (482 mg,
0.357 mmol, 90%) was obtained.
【００４５】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.23-1.37 (36H, m), 1.43 (4H,m), 1.48 (2H, m),
1.57 (2H, quint, J=6.8 Hz), 1.75 (4H, quint, J=6.
8 Hz), 1.93 (1H, m), 2.05 (1H, m), 2.43 (2H, m),
3.10 (1H, brs), 3.17 (2H, m), 3.44 (2H, t, J=6.6 H
z), 3.47 (1H, quint, J=1.7 Hz), 3.72-3.85 (6H, m),
3.95 (4H, t, J=6.6 Hz), 4.01 (1H, m), 4.18 (1H, t,
J=6.8 Hz), 4.36 (1H,dd, J=10.5, 6.8 Hz), 4.43 (1
H, dd, J=10.5, 6.8 Hz), 6.55 (1H, t, J=2.2Hz), 6.8
7 (2H, d, J=2.2 Hz), 6.98 (1H, brs), 7.11 (1H, br
s), 7.23 (18H,m), 7.38 (2H, t, J=7.6 Hz), 7.58 (3
H, m), 7.80 (2H, d, J=7.6 Hz).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.23-1.37 (36H, m), 1.43 (4H, m), 1.48 (2H, m),
1.57 (2H, quint, J = 6.8 Hz), 1.75 (4H, quint, J = 6.
8 Hz), 1.93 (1H, m), 2.05 (1H, m), 2.43 (2H, m),
3.10 (1H, brs), 3.17 (2H, m), 3.44 (2H, t, J = 6.6 H
z), 3.47 (1H, quint, J = 1.7 Hz), 3.72-3.85 (6H, m),
3.95 (4H, t, J = 6.6 Hz), 4.01 (1H, m), 4.18 (1H, t,
J = 6.8 Hz), 4.36 (1H, dd, J = 10.5, 6.8 Hz), 4.43 (1
H, dd, J = 10.5, 6.8 Hz), 6.55 (1H, t, J = 2.2Hz), 6.8
7 (2H, d, J = 2.2 Hz), 6.98 (1H, brs), 7.11 (1H, br
s), 7.23 (18H, m), 7.38 (2H, t, J = 7.6 Hz), 7.58 (3
H, m), 7.80 (2H, d, J = 7.6 Hz).
【００４６】Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）（Ｆｍｏｃ）-
Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビ
ス（ドデシロキシ）ベンズアミド (400 mg, 0.296 mmo
l) をジクロロメタン (2 ml) に溶解し、ピペリジン (1
ml) を加えて、2時間撹拌した。反応溶液をＳｅｐｈａ
ｄｅｘ ＬＨ-２０（ジクロロメタン：メタノール＝
７：３）で精製し、Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）-Ｇｌｙ-
Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス（ド
デシロキシ）ベンズアミド (333 mg, 0.284 mmol, 96%)
を得た。N- (N-Gln (Trt) (Fmoc)-
Gly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (400 mg, 0.296 mmo
l) in dichloromethane (2 ml) and piperidine (1 ml)
ml) was added and the mixture was stirred for 2 hours. The reaction solution is Sepha
dex LH-20 (dichloromethane: methanol =
7: 3), N- (N-Gln (Trt) -Gly-
Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (333 mg, 0.284 mmol, 96%)
Got
【００４７】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.25-1.52 (42H, m), 1.58 (2H,m), 1.76 (4H, qui
nt, J=7.1 Hz), 1.81 (1H, m), 1.94 (1H, m), 2.41 (1
H, quint, J=7.3 Hz), 2.49 (1H, quint, J=7.3 Hz),
3.17 (2H, t, J=6.8 Hz), 3.27(1H, t, J=6.4 Hz), 3.3
5 (2H, t, J=6.8 Hz), 3.68-3.90 (6H. m), 3.96 (4H,
t, J=6.6 Hz), 6.56 (1H, t, J=2.2 Hz), 6.89 (2H, d,
J=2.2 Hz), 7.17-7.29(16H, m), 7.34 (1H, brs), 7.8
3 (1H, brs), 8.11 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.25-1.52 (42H, m), 1.58 (2H, m), 1.76 (4H, qui
nt, J = 7.1 Hz), 1.81 (1H, m), 1.94 (1H, m), 2.41 (1
H, quint, J = 7.3 Hz), 2.49 (1H, quint, J = 7.3 Hz),
3.17 (2H, t, J = 6.8 Hz), 3.27 (1H, t, J = 6.4 Hz), 3.3
5 (2H, t, J = 6.8 Hz), 3.68-3.90 (6H. M), 3.96 (4H,
t, J = 6.6 Hz), 6.56 (1H, t, J = 2.2 Hz), 6.89 (2H, d,
J = 2.2 Hz), 7.17-7.29 (16H, m), 7.34 (1H, brs), 7.8
3 (1H, brs), 8.11 (1H, brs).
【００４８】Ｆｍｏｃ-Ｐｒｏ-ＯＨ (111mg, 0.329 mmo
l)、水溶性カルボジイミド (63 mg,0.329 mmol)、１-ヒ
ドロキシベンゾトリアゾール (51 mg, 0.329 mmol) を
乾燥ジクロロメタンに溶解し、室温で1時間撹拌した
後、（Ｎ-（Ｎ-Ｇｌｎ（Ｔｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌ
ｙ-６-アミノヘキシル）-３，５-ビス（ドデシロキシ）
ベンズアミド (257 mg, 0.219 mmol) のジクロロメタン
溶液を加え、更に室温で２時間撹拌した。反応終了後、
反応溶液を水洗し、硫酸マグネシウムで乾燥させた。硫
酸マグネシウムをろ別し、溶媒を減圧下留去した。残渣
をシリカゲルカラムクロマトグラフィー（クロロホル
ム：メタノール＝９８：２）で精製し、Ｎ-（Ｎ-Ｐｒｏ
（Ｆｍｏｃ）-Ｇｌｎ（Ｔｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ
-６-アミノヘキシル）-３，５-ビス（ドデシロキシ）ベ
ンズアミド (301 mg, 0.208 mmol,95%) を得た。Fmoc-Pro-OH (111 mg, 0.329 mmo
l), water-soluble carbodiimide (63 mg, 0.329 mmol), 1-hydroxybenzotriazole (51 mg, 0.329 mmol) were dissolved in dry dichloromethane, and the mixture was stirred at room temperature for 1 hour, and then (N- (N-Gln (Trt ) -Gly-Gly-Gl
y-6-aminohexyl) -3,5-bis (dodecyloxy)
A dichloromethane solution of benzamide (257 mg, 0.219 mmol) was added, and the mixture was further stirred at room temperature for 2 hours. After the reaction,
The reaction solution was washed with water and dried over magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 98: 2), and N- (N-Pro
(Fmoc) -Gln (Trt) -Gly-Gly-Gly
-6-Aminohexyl) -3,5-bis (dodecyloxy) benzamide (301 mg, 0.208 mmol, 95%) was obtained.
【００４９】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.20-1.70 (42H, m), 1.74 (6H,m), 1.96-2.21 (4
H, m), 2.44 (1H, dd, J=16.1, 7.3 Hz), 2.72 (1H, d
d, J=16.1, 7.3 Hz), 3.03 (1H, brs), 3.13 (2H, q, J
=6.3 Hz), 3.32 (3H, m), 3.52(1H, dd, J=16.8 Hz),
3.74 (1H, dd, J=16.8, 5.1 Hz), 4.9 Hz), 3.80 (2H,
m), 3.92 (4H, t, J=6.6 Hz), 3.93-4.12 (8H, m), 4.3
0 (1H, dd, J=9.8, 6.6 Hz), 6.53 (1H, t, J=2.2 Hz),
6.56 (1H, t, J=5.6 Hz), 6.79 (1H, t, J=5.6 Hz),
6.85 (2H, d, J=2.2 Hz), 6.90 (1H, brs), 7.15 (15H,
m), 7.42 (6H, m),7.61 (1H, t, J=6.1 Hz), 7.77 (2
H, d, J=7.6 Hz), 7.92 (1H, brs), 9.45 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.20-1.70 (42H, m), 1.74 (6H, m), 1.96-2.21 (4
H, m), 2.44 (1H, dd, J = 16.1, 7.3 Hz), 2.72 (1H, d
d, J = 16.1, 7.3 Hz), 3.03 (1H, brs), 3.13 (2H, q, J
= 6.3 Hz), 3.32 (3H, m), 3.52 (1H, dd, J = 16.8 Hz),
3.74 (1H, dd, J = 16.8, 5.1 Hz), 4.9 Hz), 3.80 (2H,
m), 3.92 (4H, t, J = 6.6 Hz), 3.93-4.12 (8H, m), 4.3
0 (1H, dd, J = 9.8, 6.6 Hz), 6.53 (1H, t, J = 2.2 Hz),
6.56 (1H, t, J = 5.6 Hz), 6.79 (1H, t, J = 5.6 Hz),
6.85 (2H, d, J = 2.2 Hz), 6.90 (1H, brs), 7.15 (15H,
m), 7.42 (6H, m), 7.61 (1H, t, J = 6.1 Hz), 7.77 (2
H, d, J = 7.6 Hz), 7.92 (1H, brs), 9.45 (1H, brs).
【００５０】Ｎ-（Ｎ-Ｐｒｏ（Ｆｍｏｃ）-Ｇｌｎ（Ｔ
ｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミド (250 mg,
0.173 mmol) をジクロロメタン (2 ml) に溶解し、ピペ
リジン (1 ml) を加えて、2時間撹拌した。反応溶液を
Ｓｅｐｈａｄｅｘ ＬＨ-２０（ジクロロメタン：メタ
ノール＝７：３）で精製し、Ｎ-（Ｎ-Ｐｒｏ-Ｇｌｎ
（Ｔｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシ
ル）-３，５-ビス（ドデシロキシ）ベンズアミド (204
mg, 0.166 mmol, 96%) を得た。N- (N-Pro (Fmoc) -Gln (T
rt) -Gly-Gly-Gly-6-aminohexyl)-
3,5-bis (dodecyloxy) benzamide (250 mg,
0.173 mmol) was dissolved in dichloromethane (2 ml), piperidine (1 ml) was added, and the mixture was stirred for 2 hours. The reaction solution was purified with Sephadex LH-20 (dichloromethane: methanol = 7: 3), and N- (N-Pro-Gln was used.
(Trt) -Gly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (204
mg, 0.166 mmol, 96%) was obtained.
【００５１】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.23-1.65 (40H, m), 1.49 (9H,m), 1.95 (1H, q,
J=7.3 Hz), 2.09 (2H, m), 2.46 (2H, m), 2.98 (2H,
t, J=6.6 Hz), 3.18 (2H, t, J=6.6 Hz), 3.35 (4H,
m), 3.66-3.83 (7H, m), 3.95 (4H, t, J=6.6 Hz), 4.1
7 (1H, t, J=6.6 Hz), 6.56 (1H, t, J=2.2 Hz), 6.88
(2H, d, J=2.2 Hz), 7.16-7.30 (15H, m).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.23-1.65 (40H, m), 1.49 (9H, m), 1.95 (1H, q,
J = 7.3 Hz), 2.09 (2H, m), 2.46 (2H, m), 2.98 (2H,
t, J = 6.6 Hz), 3.18 (2H, t, J = 6.6 Hz), 3.35 (4H,
m), 3.66-3.83 (7H, m), 3.95 (4H, t, J = 6.6 Hz), 4.1
7 (1H, t, J = 6.6 Hz), 6.56 (1H, t, J = 2.2 Hz), 6.88
(2H, d, J = 2.2 Hz), 7.16-7.30 (15H, m).
【００５２】Ｆｍｏｃ-（Ｔｒｔ）Ｈｉｓ-ＯＨ (136 m
g, 0.220 mmol)、水溶性カルボジイミド (42 mg, 0.220
mmol)、１-ヒドロキシベンゾトリアゾール (34 mg, 0.
220mmol) を乾燥ジクロロメタンに溶解し、室温で1時間
撹拌した後、Ｎ-（Ｎ-Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-Ｇｌｙ
-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス（ド
デシロキシ）ベンズアミド (180 mg, 0.147 mmol) のジ
クロロメタン溶液を加え、更に室温で２時間撹拌した。
反応終了後、反応溶液を水洗し、硫酸マグネシウムで乾
燥させた。硫酸マグネシウムをろ別し、溶媒を減圧下留
去した。残渣をシリカゲルカラムクロマトグラフィー
（クロロホルム：メタノール＝９８：２）で精製し、Ｎ
-（Ｎ-Ｈｉｓ（Ｔｒｔ）（Ｆｍｏｃ）-Ｐｒｏ-Ｇｌｎ
（Ｔｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシ
ル）-３，５-ビス（ドデシロキシ）ベンズアミド (262
mg, 0.143 mmol, 97%) を得た。Fmoc- (Trt) His-OH (136 m
g, 0.220 mmol), water-soluble carbodiimide (42 mg, 0.220
mmol), 1-hydroxybenzotriazole (34 mg, 0.1.
220 mmol) was dissolved in dry dichloromethane and stirred at room temperature for 1 hour, and then N- (N-Pro-Gln (Trt) -Gly
A dichloromethane solution of -Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (180 mg, 0.147 mmol) was added, and the mixture was further stirred at room temperature for 2 hours.
After the reaction was completed, the reaction solution was washed with water and dried with magnesium sulfate. Magnesium sulfate was filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 98: 2), N
-(N-His (Trt) (Fmoc) -Pro-Gln
(Trt) -Gly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (262
mg, 0.143 mmol, 97%) was obtained.
【００５３】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.25-1.57 (44H, m), 1.75 (7H,m), 2.05 (2H, m),
2.21 (1H, m), 2.39 (1H, m), 2.50 (1H, m), 3.17 (2
H, t,J=6.3 Hz), 3.34 (2H, t, J=6.3 Hz), 3.38 (2H,
m), 3.46-3.65 (3H, m), 3.75-3.88 (6H, m), 3.95 (4
H, t, J=6.6 Hz), 4.05-4.20 (4H, m), 4.29 (2H, m),
4.38 (1H, brs), 4.48 (1H, brs), 6.54 (1H, s), 6.66
(1H, brs), 6.76 (1H,brs), 6.87 (2H, s), 7.07-7.24
(25H, m), 7.25-7.42 (9H, m), 7.54 (2H, t,J=7.6 H
z), 7.73 (2H, d, J=7.6 Hz), 8.03 (1H, brs), 8.46
(1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.25-1.57 (44H, m), 1.75 (7H, m), 2.05 (2H, m),
2.21 (1H, m), 2.39 (1H, m), 2.50 (1H, m), 3.17 (2
H, t, J = 6.3 Hz), 3.34 (2H, t, J = 6.3 Hz), 3.38 (2H,
m), 3.46-3.65 (3H, m), 3.75-3.88 (6H, m), 3.95 (4
H, t, J = 6.6 Hz), 4.05-4.20 (4H, m), 4.29 (2H, m),
4.38 (1H, brs), 4.48 (1H, brs), 6.54 (1H, s), 6.66
(1H, brs), 6.76 (1H, brs), 6.87 (2H, s), 7.07-7.24
(25H, m), 7.25-7.42 (9H, m), 7.54 (2H, t, J = 7.6 H
z), 7.73 (2H, d, J = 7.6 Hz), 8.03 (1H, brs), 8.46
(1H, brs).
【００５４】Ｎ-（Ｎ-Ｈｉｓ（Ｔｒｔ）（Ｆｍｏｃ）-
Ｐｒｏ-Ｇｌｎ（Ｔｒｔ）-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-ア
ミノヘキシル）-３，５-ビス（ドデシロキシ）ベンズア
ミド(70 mg, 0.038 mmol) にトリフルオロ酢酸 (1.0 m
l) を加えて、2時間撹拌した。トリフルオロ酢酸を減圧
下留去し、シリカゲルカラムクロマトグラフィー（クロ
ロホルム：メタノール＝８：２）で精製し、Ｎ-（Ｎ-Ｈ
ｉｓ（Ｆｍｏｃ）-Ｐｒｏ-Ｇｌｎ-Ｇｌｙ-Ｇｌｙ-Ｇｌ
ｙ-６-アミノヘキシル）-３，５-ビス（ドデシロキシ）
ベンズアミド (44 mg, 0.033 mmol, 85%) を得た。N- (N-His (Trt) (Fmoc)-
Pro-Gln (Trt) -Gly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (70 mg, 0.038 mmol) in trifluoroacetic acid (1.0 m
l) was added and stirred for 2 hours. Trifluoroacetic acid was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 8: 2), N- (NH)
is (Fmoc) -Pro-Gln-Gly-Gly-Gl
y-6-aminohexyl) -3,5-bis (dodecyloxy)
Benzamide (44 mg, 0.033 mmol, 85%) was obtained.
【００５５】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.26-1.35 (38H, m), 1.40-1.58(6H, m), 1.76 (4
H, quint, J=6.8 Hz), 2.02 (5H, m), 2.26 (1H, brs),
2.34(2H, t, J=6.8 Hz), 3.18 (2H, t, J=6.8 Hz), 3.
25 (2H, m), 3.35 (2H, t, J=6.8 Hz), 3.38 (2H, m),
3.66 (2H, m), 3.68-3.94 (6H, m), 3.96 (4H, t, J=6.
6 Hz), 4.18 (1H, t, J=6.6 Hz), 4.30 (1H, t, J=6.6
Hz), 4.34 (1H, brs),4.45 (2H, m), 4.75 (1H, brs),
6.56 (1H, t, J=2.2 Hz), 6.87 (2H, d, J=2.2Hz), 7.0
6 (1H, s), 7.30 (2H, d, J=7.6 Hz), 7.40 (2H, t, J=
7.6 Hz), 7.57(2H, d, J=7.6 Hz), 7.76 (2H, d, J=7.6
Hz), 8.60 (1H, s).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.26-1.35 (38H, m), 1.40-1.58 (6H, m), 1.76 (4
H, quint, J = 6.8 Hz), 2.02 (5H, m), 2.26 (1H, brs),
2.34 (2H, t, J = 6.8 Hz), 3.18 (2H, t, J = 6.8 Hz), 3.
25 (2H, m), 3.35 (2H, t, J = 6.8 Hz), 3.38 (2H, m),
3.66 (2H, m), 3.68-3.94 (6H, m), 3.96 (4H, t, J = 6.
6 Hz), 4.18 (1H, t, J = 6.6 Hz), 4.30 (1H, t, J = 6.6
Hz), 4.34 (1H, brs), 4.45 (2H, m), 4.75 (1H, brs),
6.56 (1H, t, J = 2.2 Hz), 6.87 (2H, d, J = 2.2Hz), 7.0
6 (1H, s), 7.30 (2H, d, J = 7.6 Hz), 7.40 (2H, t, J =
7.6 Hz), 7.57 (2H, d, J = 7.6 Hz), 7.76 (2H, d, J = 7.6
Hz), 8.60 (1H, s).
【００５６】Ｎ-（Ｎ-Ｈｉｓ（Ｆｍｏｃ）-Ｐｒｏ-Ｇｌ
ｎ-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５
-ビス（ドデシロキシ）ベンズアミド (15 mg, 0.011 mm
ol)をジクロロメタン (2 ml) に溶解し、ピペリジン (1
ml) を加えて、2時間撹拌した。反応溶液をＳｅｐｈａ
ｄｅｘ ＬＨ-２０（ジクロロメタン：メタノール＝
７：３）で精製し、Ｎ-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-Ｇ
ｌｙ-Ｇｌｙ-Ｇｌｙ-６-アミノヘキシル）-３，５-ビス
（ドデシロキシ）ベンズアミド (10 mg, 0.009mmol, 83
%) を得た。N- (N-His (Fmoc) -Pro-Gl
n-Gly-Gly-Gly-6-aminohexyl) -3,5
-Bis (dodecyloxy) benzamide (15 mg, 0.011 mm
ol) in dichloromethane (2 ml) and piperidine (1 ml
ml) was added and the mixture was stirred for 2 hours. The reaction solution is Sepha
dex LH-20 (dichloromethane: methanol =
7: 3), N- (N-His-Pro-Gln-G
ly-Gly-Gly-6-aminohexyl) -3,5-bis (dodecyloxy) benzamide (10 mg, 0.009mmol, 83
%)
【００５７】1H NMR (CDCl3)δ0.88 (6H, t, J=6.8 H
z), 1.22-1.58 (44H, m), 1.77 (4H,q, J=6.8 Hz), 1.9
4 (2H, m), 2.14 (1H, quint, J=7.3 Hz), 2.20 (1H, q
uint,J=7.3 Hz), 2.33 (1H, quint, J=7.3 Hz), 2.39
(1H, quint, J=7.3 Hz), 3.07(3H, m), 3.18 (2H, m),
3.36 (2H, q, J=7.3 Hz), 3.83-4.02 (6H, m), 3.96(4
H, t, J=6.8 Hz), 4.07 (1H, dd, J=7.3, 5.3 Hz), 4.3
7 (2H, m), 6.17 (1H,brs), 6.56 (1H, t, J=2.2 Hz),
6.90 (2H, d, J=2.2 Hz), 7.01 (1H, s), 7.28 (1H, br
s), 7.78 (1H, s), 7.90 (1H, brs), 8.01 (1H, brs),
8.20 (1H, brs).1 H NMR (CDCl 3 ) δ 0.88 (6H, t, J = 6.8 H
z), 1.22-1.58 (44H, m), 1.77 (4H, q, J = 6.8 Hz), 1.9
4 (2H, m), 2.14 (1H, quint, J = 7.3 Hz), 2.20 (1H, q
uint, J = 7.3 Hz), 2.33 (1H, quint, J = 7.3 Hz), 2.39
(1H, quint, J = 7.3 Hz), 3.07 (3H, m), 3.18 (2H, m),
3.36 (2H, q, J = 7.3 Hz), 3.83-4.02 (6H, m), 3.96 (4
H, t, J = 6.8 Hz), 4.07 (1H, dd, J = 7.3, 5.3 Hz), 4.3
7 (2H, m), 6.17 (1H, brs), 6.56 (1H, t, J = 2.2 Hz),
6.90 (2H, d, J = 2.2 Hz), 7.01 (1H, s), 7.28 (1H, br
s), 7.78 (1H, s), 7.90 (1H, brs), 8.01 (1H, brs),
8.20 (1H, brs).
【００５８】[0058]
【実施例３】ビオチン様ペプチド脂質とストレプトアビ
ジンとの相互作用
Ｎ-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-６-アミノヘキシル）-
３，５-ビス（ドデシロキシ）ベンズアミドとストレプ
トアビジンとの相互作用は、分子間相互作用解析装置で
あるＩＡｓｙｓ Ｐｌｕｓ（Ａｆｆｉｎｉｔｙ Ｓｅｎ
ｓｏｒｓ社製）を用いて解析した。Example 3 Interaction between biotin-like peptide lipid and streptavidin N- (N-His-Pro-Gln-6-aminohexyl)-
The interaction between 3,5-bis (dodecyloxy) benzamide and streptavidin is IAsys Plus (Affinity Sen) which is an intermolecular interaction analyzer.
(made by sors).
【００５９】疎水性キュベットを、界面活性剤、バッフ
ァー、２-プロパノールで洗浄の後、８μＭの試料溶液
（２-プロパノール：クロロホルム１９：１）を添加し
固定化を行った。固定化量は４９３ Ａｒｃ ｓｅｃｏ
ｎｄｓ だった。このキュベットを、バッファー、塩酸
水溶液、水酸化ナトリウム水溶液で洗浄の後、牛血清ア
ルブミンでブロッキングを行い、バッファーで洗浄の
後、ストレプトアビジン３．０ ｍｇ／ｍｌを５０μｌ
添加して、相互作用を検討した。５分後の吸着量は３０
０ Ａｒｃ ｓｅｃｏｎｄｓだった。さらに解離バッフ
ァーで置換して１０分後の吸着量を測定したところ１５
５ Ａｒｃ ｓｅｃｏｎｄｓであり、ストレプトアビジ
ンを吸着後、徐放出来ることを確認した。The hydrophobic cuvette was washed with a surfactant, a buffer and 2-propanol, and then an 8 μM sample solution (2-propanol: chloroform 19: 1) was added for immobilization. Immobilization amount is 493 Arc seco
It was nds. The cuvette was washed with a buffer, an aqueous solution of hydrochloric acid and an aqueous solution of sodium hydroxide, blocked with bovine serum albumin, washed with a buffer, and then 50 μl of streptavidin 3.0 mg / ml.
Addition and interaction was investigated. Adsorption amount after 5 minutes is 30
It was 0 Arc seconds. Further, the amount of adsorption was measured after 10 minutes of replacement with the dissociation buffer.
It was 5 Arc seconds, and it was confirmed that streptavidin could be adsorbed and then gradually released.
【００６０】[0060]
【実施例４】ビオチン様ペプチド脂質とストレプトアビ
ジンとの相互作用
Ｎ-（Ｎ-Ｈｉｓ-Ｐｒｏ-Ｇｌｎ-Ｇｌｙ-Ｇｌｙ-Ｇｌｙ-
６-アミノヘキシル）-３，５-ビス（ドデシロキシ）ベ
ンズアミドとストレプトアビジンとの相互作用の解析を
行った。０．８μＭの試料溶液（２-プロパノール：ク
ロロホルム１９：１）を添加して固定化を行った。固定
化量は９４７ Ａｒｃ ｓｅｃｏｎｄｓだった。ストレ
プトアビジン３．０ ｍｇ／ｍｌを５０μｌ添加後、５
分後の吸着量は４５０ Ａｒｃ ｓｅｃｏｎｄｓだっ
た。さらに解離バッファーで置換して１０分後の吸着量
を測定したところ２８０ Ａｒｃ ｓｅｃｏｎｄｓであ
り、ストレプトアビジンを吸着後、徐放出来ることを確
認した。Example 4 Interaction between biotin-like peptide lipid and streptavidin N- (N-His-Pro-Gln-Gly-Gly-Gly-
The interaction between 6-aminohexyl) -3,5-bis (dodecyloxy) benzamide and streptavidin was analyzed. Immobilization was performed by adding 0.8 μM sample solution (2-propanol: chloroform 19: 1). The immobilized amount was 947 Arc seconds. After adding 50 μl of 3.0 mg / ml streptavidin, 5
The adsorbed amount after 450 minutes was 450 Arc seconds. Further, the amount of adsorption was measured after 10 minutes after substitution with the dissociation buffer, and it was 280 Arc seconds, and it was confirmed that streptavidin could be adsorbed and then gradually released.
【００６１】[0061]
【発明の効果】本発明は、疎水性表面にコーティングす
ることにより、細胞培養等に使用するタンパク質性の因
子を、材料表面から徐放する機能を付与する新規な構造
を有するビオチン様ペプチド脂質を提供することにあ
る。INDUSTRIAL APPLICABILITY The present invention provides a biotin-like peptide lipid having a novel structure, which is coated on a hydrophobic surface to impart a function of gradually releasing a proteinaceous factor used for cell culture or the like from the material surface. To provide.
Claims (4)
Ｒ1を表し、ｌは０から３の整数を、ｍは２から６の整
数を、ｎは１１から１７の整数を、Ｈｉｓはヒスチジン
を、Ｐｒｏはプロリンを、Ｇｌｎはグルタミンを、Ｇｌ
ｙはグリシンを表わす。）1. A peptide lipid represented by the formula (1). [Chemical 1] (In the formula, R 1 represents an O (CH 2 ) n CH 3 group, R 2 represents H or R 1 , l is an integer of 0 to 3, m is an integer of 2 to 6, and n is 11 to An integer of 17, His for histidine, Pro for proline, Gln for glutamine and Gl.
y represents glycine. )
パク質徐放素材。2. A protein sustained-release material having a His-Pro-Gln structure.
成分とするタンパク質徐放素材。3. A protein sustained-release material containing a peptide lipid represented by the formula (1) as an active ingredient.
成分とするタンパク質徐放剤。4. A sustained-release agent for proteins, which comprises a peptide lipid represented by the formula (1) as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002060652A JP2003261595A (en) | 2002-03-06 | 2002-03-06 | Biotin-like peptide lipid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002060652A JP2003261595A (en) | 2002-03-06 | 2002-03-06 | Biotin-like peptide lipid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2003261595A true JP2003261595A (en) | 2003-09-19 |
Family
ID=29195604
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2002060652A Pending JP2003261595A (en) | 2002-03-06 | 2002-03-06 | Biotin-like peptide lipid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2003261595A (en) |
-
2002
- 2002-03-06 JP JP2002060652A patent/JP2003261595A/en active Pending
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