고분자에 결합된 활성 에스테르에 의한 펩티드합성 III. 1-페닐-3-메틸-4-옥시미노피라졸레진을 이용한 펩티드 합성에서 Spacer Arm의 효과

왕영; 이윤식

Journal of the Korean Industrial and Engineering Chemistry, Vol.4, No.1, 132-143, March, 1993

Peptide Synthesis with Polymer Bound Active Ester Ⅲ. The Effect of Spacer Arm in Peptide Synthesis with 1-Phenyl-3-methyl-4-oximino pyrazole Resin

왕영, 이윤식

초록

Aminomethyl레진을 chloromethyl레진(Merrifield레진)으로부터 또는 polystyrene레진을 직접 amidoalkylation하여 각각 합성하였다. 두 종류의 aminomethyl레진을 이용하여 5개의 ε-aminocaproic acid(ACA)가 차례로 커플링 된 spacer arm을 가진 레진들을 각각 합성하였다. Chloromethyl레진으로부터 합성된 aminomethyl레진의 경우 ACA를 매번 커플링 할 때마다 25∼30%의 유리 아미노기의 양이 감소하였으며, 직접 amidoalkylation에 의해 합성한 amlnomethyl 레진의 경우 매 커플링 단계 마다 3∼5%의 유리 아미노기의 양이 감소하였다. 4-Nitroso-5-aminopyrazole 기능기를 가진 레진은 직접 amidoalkylation하여 얻은 레진에 ACA를 spacer arm으로 커플링시켜 얻은 레진과 5-phenyl-7-methylpyrazole(4,3-c)(1,2,4)oxadiazin-3-one을 반응시켜 얻었다. 4-Nitroso-5-aminopyrazole 기능기를 가진 레진을 이용하여 α-아미노기가 보호된 여러 가지 아미노산의 활성에스테르 레진들을 합성하였다. 4-Nitroso-5-aminopyrazole 기능기를 가진 활성 에스테르 레진은 N-acylation 반응에 매우 뛰어난 반응성을 나타내었다. 또한 입체장애 효과 없이 아미노산 유도체의 종류에 거의 무관하게 아실화 반응이 일어났으며 90∼96%의 수율로 펩티드들을 합성할 수 있었다. 얻어진 펩티드들은 NMR을 비롯한 여러 가지 물리적 방법으로 그 순도를 확인하였다.

Aminomethyl polystyrene resins were prepared either from chloromethyl-resin(Merrifield resin) or from direct amidoalkylation of polystyrene resin. Two kinds of aminomethlyl resin were lengthened with spacer arms via sequential coupling of five ε-aminocaproic acids(ACA) respectively. In case of the resin prepared from the Merrifield resin, the amounts of free amino group of the resin were reduced by 25∼30% after each coupling of ACA. But the one from direct amidoalkylation showed 3∼5% loss after each coupling of ACA. 4-Nitroso-5-aminopyrazole resin was made by reacting ACA spacer arm resin, which was made from direct amidoalkylated resin, with 5-phenyl-7-methylpyrazole [4,3-c][1,2,4]oxadiazin-3-one. Several polymeric active esters of N-blocked amino acids were prepared from the 4-nitroso-5-aminopyrazole bound resins. In anchoring step of the amino acid derivatives on the resin, no substantial effect of bulkiness was found. 4-Nitroso-5-aminopyrazole bound active ester resins were found to be very reactive in N-acylation, The resulting peptides were obtained with 90∼95% yield and characterized by NMR and other physical methods.

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[1] Merrifield RB, J. Am. Chem. Soc., 85, 2147 (1963)

[2] Degrado WF, Kaiser ET, J. Org. Chem., 45, 1295 (1980)

[3] Degrado WF, Kaiser ET, J. Org. Chem., 47, 3258 (1982)

[4] Nakagawa SH, Kaiser ET, J. Org. Chem., 48, 678 (1983)

[5] Fridkin M, Stabinsky Y, Zakuth V, Spirer Z, Biochim. Biophys. Acta, 49, 203 (1977)

[6] Tam JP, Tjoeng FS, Merrifield RB, J. Am. Chem. Soc., 105, 6117 (1980)

[7] Tam JP, Tjoeng FS, Merrifield RB, Tetrahedron Lett., 31, 6493 (1980)

[8] Tam JP, Tjoeng FS, Merrifield RB, Int. J. Pep. Protein Res., 16, 412 (1980)

[9] Kim JB, Lee YS, Bull. Korean Chem. Soc., 12, 376 (1991)

[10] Lee KW, Lee YS, Bull. Korean Chem. Soc., 10, 331 (1989)

[11] RaChemle JR, J. Org. Chem., 28, 3898 (1963)

[12] Zervas L, Winitz M, Greenstein JP, J. Org. Chem., 22, 1515 (1955)

[13] Gisin BF, Anal. Chim. Acta, 58 (1972)

[14] Stewart JM, Young JD, "Solid Phase Peptide Sythesis," 2nd ed., Pierce Chemical Company, Rockford, Illinois, p. 105 (1984)

[15] Stewart JM, Yonng JD, "Solid Phase Peptide Sythesis," 2nd ed., Pierce Chemical Company, Rockford, Illinois, p. 113 (1984)

[16] Kalir R, Fridkin N, Patchornik A, Eur. J. Biochem., 42, 151 (1974)

[17] Mitchell AR, Merrifield RB, J. Org. Chem., 43, 2845 (1978)

[18] Kent SBH, Mitchemll AR, Engelhard M, Merrifield RB, Proc. Natl. Acad. Sci. U.S.A., 76, 2180 (1978)

[19] Ondetti MA, Williams NJ, Sabo EF, Plusces J, Biochemistry, 10, 4033 (1971)

[20] Birr C, "Aspects of the Merrifield Peptide Synthesis," In "Reactivity and Structure Concepts in Organic Chemistry," (K. Hafner et al. eds.), Vol. 8, Springer-Verlag, Berlin, p. 42 (1978)

[21] Taylor EC, Hartke KS, J. Am. Chem. Soc., 81, 6117 (1980)

[22] Guarneri M, Menegatti E, Giori P, Ferroni RX, Tomatis R, Benassi CA, Gazz. Chim. Ital., 103, 1105 (1973)

[23] Sparrow JT, J. Org. Chem., 41, 1350 (1976)