基于Loloatin C的系列假环多肽的设计合成及抑菌机理的初步研究
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摘要
Loloatin C是从热带海洋细菌的实验室培养物中发现、分离得到的环十肽化合物,大量实验结果显示它对革兰氏阳性菌及阴性菌均表现出很好的活性。前人通过滑动图窗的方法将Loloatin C分为南、中、北三个环六肽,成功合成得到环肽,并进一步进行药敏实验,结果发现药核是包含–D-Tyr-Pro-Trp-D-Phe-片段的环状化合物。
我们采用固相及液相合成法,设计并合成一系列包含–D-Tyr-Pro-Trp-D-Phe-片段的假环肽化合物,期望得到活性更好的假环肽化合物。
为了初步了解Loloatin C的抑菌机制,我们研究了未经处理及在CCCP预处理情况下Loloatin C对大肠杆菌生长的抑制情况,以及以乳糖为诱导剂作用下大肠杆菌的β-半乳糖苷酶的活性。
研究结果:(1)成功合成5个线性多肽前体(S1-1、S1-2、S、S2-1、S2-2)及3个假环肽化合物(C1-2、C2-1、C2-2),线性多肽前体的总产率在40%左右;而假环肽的总产率较低,大约为2%左右;(2)大肠杆菌在CCCP的作用下其存活率明显提高,从而推断Loloatin C抗菌肽不会因致病细菌的主动外排系统而产生耐药性的问题。(3)实验菌株大肠杆菌的β-半乳糖苷酶活性随着多肽药物Loloatin C的作用浓度的增加而相应地增强。
Loloatin C was discovered and separated from tropical ocean bacterial laboratory, which is a cyclodecapeptide compound. Many experiments have shown that Loloatin C has remarkable activity to G+ and G- bacteria. Three cyclohexapeptides (named north, middle, and south cyclohexapeptide) narrowed down from Loloatin C by sliding window were figured out and synthesized. Antimicrobial screening conformed that the pharmacophore of Loloatin C contains structure of–D-Tyr-Pro-Trp-D-Phe-. Based on these results, we design and synthesize series of pseudocyclicpeptides, which contain–D-Tyr-Pro-Trp-D-Phe-, expecting to get more active compounds.
In order to study the antibacterial mechanism of Loloatin C, growth inhibition of loloatin C to E. coli was investigated under the existence or without the existence of CCCP. The activity ofβ-galactose glycosidase when Loloatin C interacted with E. coli was also studied by using lactose as substrate.
The following results have been obtained: (1) Five linear precursor peptides (S1-1, S1-2, S, S2-1, S2-2)were synthesized and the overall yields were about 40%; three pseudo cyclopeptides (C1-2, C2-1, C2-2) were obtained but the overall yield were low, only 2%; (2) More E.coli bacteria survive under the existence of CCCP, which we can infer that the problem of resistance won’t happen due to the excessive expression of efflux system of bacteria. Besides, as the concentration of Loloatin C increases, the activity ofβ-galactose glycoside from E.coli gets stronger.
我们采用固相及液相合成法,设计并合成一系列包含–D-Tyr-Pro-Trp-D-Phe-片段的假环肽化合物,期望得到活性更好的假环肽化合物。
为了初步了解Loloatin C的抑菌机制,我们研究了未经处理及在CCCP预处理情况下Loloatin C对大肠杆菌生长的抑制情况,以及以乳糖为诱导剂作用下大肠杆菌的β-半乳糖苷酶的活性。
研究结果:(1)成功合成5个线性多肽前体(S1-1、S1-2、S、S2-1、S2-2)及3个假环肽化合物(C1-2、C2-1、C2-2),线性多肽前体的总产率在40%左右;而假环肽的总产率较低,大约为2%左右;(2)大肠杆菌在CCCP的作用下其存活率明显提高,从而推断Loloatin C抗菌肽不会因致病细菌的主动外排系统而产生耐药性的问题。(3)实验菌株大肠杆菌的β-半乳糖苷酶活性随着多肽药物Loloatin C的作用浓度的增加而相应地增强。
Loloatin C was discovered and separated from tropical ocean bacterial laboratory, which is a cyclodecapeptide compound. Many experiments have shown that Loloatin C has remarkable activity to G+ and G- bacteria. Three cyclohexapeptides (named north, middle, and south cyclohexapeptide) narrowed down from Loloatin C by sliding window were figured out and synthesized. Antimicrobial screening conformed that the pharmacophore of Loloatin C contains structure of–D-Tyr-Pro-Trp-D-Phe-. Based on these results, we design and synthesize series of pseudocyclicpeptides, which contain–D-Tyr-Pro-Trp-D-Phe-, expecting to get more active compounds.
In order to study the antibacterial mechanism of Loloatin C, growth inhibition of loloatin C to E. coli was investigated under the existence or without the existence of CCCP. The activity ofβ-galactose glycosidase when Loloatin C interacted with E. coli was also studied by using lactose as substrate.
The following results have been obtained: (1) Five linear precursor peptides (S1-1, S1-2, S, S2-1, S2-2)were synthesized and the overall yields were about 40%; three pseudo cyclopeptides (C1-2, C2-1, C2-2) were obtained but the overall yield were low, only 2%; (2) More E.coli bacteria survive under the existence of CCCP, which we can infer that the problem of resistance won’t happen due to the excessive expression of efflux system of bacteria. Besides, as the concentration of Loloatin C increases, the activity ofβ-galactose glycoside from E.coli gets stronger.
引文
[1]. Steiner H, Hultmark D, Engstrom A, et al. Sequence and specificity of two antibacterial proteins insect immunity[J]. Nature, 1981, 292: 246~251.
[2].赵东红,戴竹英,周开亚等.昆虫抗菌肽的功能、作用机理与分子生物学研究最新进展[J].生物工程进展, 1999, 19(5): 14~18.
[3]. SCHWAB IR, DRIES D, CULLOR J, et al. Corneal storage medium preservation with efensins[J]. Comea, 1992, 11(5): 370~375.
[4].尹文,徐晨,马戈甲.抗菌肽的研究进展[J].细胞与分子免疫学, 2005, 21.
[5]. Bulet P, Urge L, Ohresser S, et al. Enlarged scale chemical synthesis and range of activity of drosocin, an O-glycosylated antibacterial peptide of Drosophila[J]. Eur J Biochem, 1996, (238): 64~69.
[6]. Tailor RH, Acland DP, Attenborough S, et al. A Novel Family of Small Cysteine-rich Antimicrobial Peptides from Seed of Impatiens balsamina Is Derived from a Single Precursor Protein[J]. J Biol Chem, 1997, 272: 24480~24887.
[7]. P. Delatorre, J. R. Olivieri, J. Ruggiero Neto, et al. Preliminary cryocrystallography analysis of an eumenine mastoparan toxin isolated from the venom of the wasp Anterhynchium flavomarginatum micado[J]. Biochim Biophys Acta, 2001, 1545: 372~376.
[8]. Clarisse Lorin, Héla Saidi, Afifa Belaid, Amira Zairi, Fran?oise Baleux, Hakim Hocini, et al. The antimicrobial peptide dermaseptin S4 inhibits HIV-1 infectivity in vitro[J]. Virology, 2005, 334: 264~275.
[9]. A. Belaid, M. Aouni, R. Khelifa, A. Trabelsi, M. Jemmali, K. Hani, et al. In vitro antiviral activity of dermaseptins against herpes simplex virus type 1[J]. Journal of Medical Virology, 2002, 66: 229~234.
[10].韩彤彤,肖向红,徐艳春.动物抗菌肽抗病毒性质及其机制[J].经济动物学报, 2006, 10 (3) : 176~179.
[11]. Marcos JF, Beachy RN, Houghten RA, et al. Inhibition of plant virus by analogs of melittin[J]. Proc Natl Acad Sci U S A, 1995, 92 (26): 12466~12469.
[12]. A. Baghian, J. Jaynes, F. Enright and K. G. Kousoulas. An Amphipathicα-Helical Synthetic Peptide Analogue of Melittin Inhibits Herpes Simplex Virus-1(HSV-1)-Induced Cell Fusion and Virus Spread[J]. Peptides, 1997, 18: 177~183.
[13]. Wachinger M, Kleinschmidt A, Winder D, et al. Antimicrobial peptides melittin and cecropin inhibit replication of human immunodeficiency virus 1 by suppressing viral gene expression[J]. J Gen Virol, 1998, 79 (4): 731~740.
[14]. JAYNES J. Lyticpeptides: A magic bullet [J]. Biotechnology News, 1990, 11(8).
[15].张卫民,彭朝晖,黄自然等.人工合成柞蚕抗菌肽对人肝癌细胞的杀伤作用[J].中华肝脏病杂志, 1998, 6(3): 158~160.
[16].张卫民,彭朝晖,黄自然等.柞蚕抗菌肽D对人直肠癌细胞的杀伤作用及机制[J].蚕业科学, 1998, 24(3): 144~149.
[17].胡云龙,天然抗菌肽分离、纯化及其抑制肿瘤细胞的作用[M].北京:中国农业科技出版社, 1996, 89~97.
[18]. Dewey G McCafferty, Predrag Cudic, Michael K Yu, Douglas C Behenna and Ryan Kruger. Synergy and duality in peptide antibiotic mechanisms[J]. Current Opinion in Chemical Biology, 1999, (3): 672~680.
[19].宋宏霞,曾名勇,刘尊英等.抗菌肽的生物活性及其作用机理[J].食品工业科技, 2006, 27(9): 185~188.
[20]. J Alberola, A Rodríguez, O Francino, X Roura, L Rivas, and D Andreu. Safety and Efficacy of Antimicrobial Peptides against Naturally Acquired Leishmaniasis. Biopolymers, 1998, 47: 415~433.
[21].卢其能,罗晓明,江辉,邹一平.抗菌肽的研究进展.江西科学, 2007, 25(4): 428~433.
[22]. Vincent du Vigneaud, Charlotte Ressler, John M Swan, Carleton W Roberts, et al. The Synthesis of Oxytocin. J Am Chem Soc, 1954, 76 : 3115~ 3121.
[23]. R B Merrifield. Solid Phase Peptide Synthesis. I. The Synthesis of a Tetrapeptide. J Am Chem Soc, 1963, 85 (14): 2149~2154.
[24].程绍玲,黄文强,何炳林.组合化学中的聚合物研究进展[J].离子交换与吸附, 1999, 15(5): 472~480.
[25]. Heru Chen, Richard K Haynes, Jürgen Scherkenbeck. Synthesis of Cyclichexapeptides using Indirect Cyclization Method [J]. Eur J Org Chem, 2004: 38~47.
[26]. Pinner R W, et al. Trends in infectious diseases mortality in the United States[J]. J Am Med Assoc, 1996, 275, 189~193.
[27]. Prenner E J, McElhaney R N, et al. The interaction of the antibacterial peptide gramicidin S with lipid bilayer model and biological membranes[J]. Biochim Biophys Acta, 1999, 1462, 201~221.
[28]. Tzong-Hsien Lee, Don Rivett, Jerome Werkmeister, Dean Hewish, Marie-Isabel Aguilar. Interaction of amphipathic peptideswith an immobilized model membrane[J]. Letters in Peptide Science, 1999, 6: 371~380.
[29]. PerezPaya E, Houghten R A, Blondelle S E. Functionalized protein-like structures from conformationally defined synthetic combinatorial libraries[J]. J Biol Chem, 1995, 270, 1048~1056.
[30]. PerezPaya E, Houghten R A, Blondelle S E. Determination of the secondary structure of selected melittin analogues with different haemolytic activities[J]. Biochem J, 1994, 299, 587~591.
[31]. Gilon C, Halle D, Chorey M, Selinger Z, Byk G. Backbone cyclization: A New Method for Conferring Conformational Constraint on Peptides[J]. Biopolymers, 1991, 31(6), 745~750.
[1]. V K Sarin, S B H Kent. Quantitative Monitoring of solid phase peptides synthesis by acid-based indicator. Coll Czech Chem Comm, 1988, 53: 2542.
[2]. Fields G B, Fields C G. Solvation effects in solid-phase peptide synthesis. J Am Chem Soc, 1991, 113: 4202~4207.
[3]. George W Anderson, Richard W Young. The Use of Diester Chlorophosphites in Peptide Syntheses. Mixed Anhydrides. J Am Chem Soc, 1952, 74: 5307~5309.
[4]. G W Anderson, F M Callahan. Racemization by the dicyclhexylcarbodiimide method of peptide synthesis. J Am Chem Soc, 1958, 80: 2902~2903.
[5]. F Weygand et al. Gaschromatographische Untersuchung der Racemisierung bei Peptidsynthesen. Angew Chem, 1963, 75 : 282~287.
[6]. Daniel S Kemp, S W Wang, G Busby III, G Hugel. Microanalysis by successive isotopic dilution. New assay for racemic content. J Am Chem Soc, 1970, 92: 1043~ 1055.
[7]. N Izumiya, M Muraoka. Racemization test in peptide synthesis. J Am Chem Soc, 1969, 91: 2391~ 2392.
[8].韩香,顾军.多肽的固相合成[J].天津药学, 2002, 14(1): 7~ 9.
[9]. ShigekiIsomura, PeterWirsching. An Immunotherapeutic Program for the Treatment of Nicotine Addiction: Hapten Design and Synthesis. J Org Chem, 2001, 66: 4115~4121.
[10]. Liren Huang, James C. Quada Jr, et al Design, Synthesis, and Sequence Selective DNA Cleavage of Functional Models of Bleomycin. 1. Hybrids Incorporating a Simple Metal-Complexing Moiety of Bleomycin and Lexitropsin Carriers[J]. Bioconjugate Chem, 1995, 6, 21-33
[11]. Ana Montero, Pilar Goya, Nadine Jagerovic. Guanidinium and Aminoimidazolinium Derivatives of N-(4-Piperidyl)Propanamides as Potential Ligands fouμOpioid and I2-imidazoline Receptors:Synthesis and Pharmacological Screening. Bioorganic and Medicinal Chemistry, 2002, 10: 1009~1018.
[12]. Mira S, Dejan M Godjevac. Synthesis and antioxidant properties of fullero-steroial covalent conjugates. Carbon, 2007, 45: 2260~2265.
[13].王德心.固相有机合成[M].北京:化学工业出版社,2004, 55-58.
[14]. Besser D, Reissmann S, Olender R, Rosenfeld R, Arad O. Study on the cyclizationtendency of backbone cyclic tetrapeptides. J Peptide Res, 2000, 56: 337-345.
[1]. Chen H R, Haynes R K, and Scherkenbeck J. Synthesis of Cyclohexapeptides Based on the Antibiotic Cyclic Decapeptide Loloatin C by an in situ Non-directed Cyclization. European J Org Chem, 2004, 1, 38～47.
[2]. Timothy J Falla, D Nedra Karunaratne and Robert E W Hancock[J]. The Journal of Biological Chemistry. 1996, 273(32): 19298～19303.
[3]. Brenwald N P, Gill M J, Wise R. Prevalence of a putative efflux mechanism among fluoroquinolone resistant clinical isolates[J]. Antimicrob Agents Chemother, 1998, 42(8): 2032～2035.
[4].王玉宝,宋诗铎,魏殿军等.多重耐药泵抑制剂提高肠球菌对氟喹诺酮药物敏感性的研究[J].中国抗生素杂志,2004,29(3):166～168.
[5].魏殿军,宋诗铎,马金玲等.泵抑制剂对耐喹诺酮类铜绿假单胞菌临床株作用的研[J].中国抗生素杂志,2004,29(8):465～468.
[6].陶传敏,李小春,吕晓菊等.氟喹诺酮类药物对嗜麦芽寡养单胞菌的抗菌活性及羰基氰氯苯腙对其的影响[J].中国抗生素杂志,2004,29(8): 469～472.
[2].赵东红,戴竹英,周开亚等.昆虫抗菌肽的功能、作用机理与分子生物学研究最新进展[J].生物工程进展, 1999, 19(5): 14~18.
[3]. SCHWAB IR, DRIES D, CULLOR J, et al. Corneal storage medium preservation with efensins[J]. Comea, 1992, 11(5): 370~375.
[4].尹文,徐晨,马戈甲.抗菌肽的研究进展[J].细胞与分子免疫学, 2005, 21.
[5]. Bulet P, Urge L, Ohresser S, et al. Enlarged scale chemical synthesis and range of activity of drosocin, an O-glycosylated antibacterial peptide of Drosophila[J]. Eur J Biochem, 1996, (238): 64~69.
[6]. Tailor RH, Acland DP, Attenborough S, et al. A Novel Family of Small Cysteine-rich Antimicrobial Peptides from Seed of Impatiens balsamina Is Derived from a Single Precursor Protein[J]. J Biol Chem, 1997, 272: 24480~24887.
[7]. P. Delatorre, J. R. Olivieri, J. Ruggiero Neto, et al. Preliminary cryocrystallography analysis of an eumenine mastoparan toxin isolated from the venom of the wasp Anterhynchium flavomarginatum micado[J]. Biochim Biophys Acta, 2001, 1545: 372~376.
[8]. Clarisse Lorin, Héla Saidi, Afifa Belaid, Amira Zairi, Fran?oise Baleux, Hakim Hocini, et al. The antimicrobial peptide dermaseptin S4 inhibits HIV-1 infectivity in vitro[J]. Virology, 2005, 334: 264~275.
[9]. A. Belaid, M. Aouni, R. Khelifa, A. Trabelsi, M. Jemmali, K. Hani, et al. In vitro antiviral activity of dermaseptins against herpes simplex virus type 1[J]. Journal of Medical Virology, 2002, 66: 229~234.
[10].韩彤彤,肖向红,徐艳春.动物抗菌肽抗病毒性质及其机制[J].经济动物学报, 2006, 10 (3) : 176~179.
[11]. Marcos JF, Beachy RN, Houghten RA, et al. Inhibition of plant virus by analogs of melittin[J]. Proc Natl Acad Sci U S A, 1995, 92 (26): 12466~12469.
[12]. A. Baghian, J. Jaynes, F. Enright and K. G. Kousoulas. An Amphipathicα-Helical Synthetic Peptide Analogue of Melittin Inhibits Herpes Simplex Virus-1(HSV-1)-Induced Cell Fusion and Virus Spread[J]. Peptides, 1997, 18: 177~183.
[13]. Wachinger M, Kleinschmidt A, Winder D, et al. Antimicrobial peptides melittin and cecropin inhibit replication of human immunodeficiency virus 1 by suppressing viral gene expression[J]. J Gen Virol, 1998, 79 (4): 731~740.
[14]. JAYNES J. Lyticpeptides: A magic bullet [J]. Biotechnology News, 1990, 11(8).
[15].张卫民,彭朝晖,黄自然等.人工合成柞蚕抗菌肽对人肝癌细胞的杀伤作用[J].中华肝脏病杂志, 1998, 6(3): 158~160.
[16].张卫民,彭朝晖,黄自然等.柞蚕抗菌肽D对人直肠癌细胞的杀伤作用及机制[J].蚕业科学, 1998, 24(3): 144~149.
[17].胡云龙,天然抗菌肽分离、纯化及其抑制肿瘤细胞的作用[M].北京:中国农业科技出版社, 1996, 89~97.
[18]. Dewey G McCafferty, Predrag Cudic, Michael K Yu, Douglas C Behenna and Ryan Kruger. Synergy and duality in peptide antibiotic mechanisms[J]. Current Opinion in Chemical Biology, 1999, (3): 672~680.
[19].宋宏霞,曾名勇,刘尊英等.抗菌肽的生物活性及其作用机理[J].食品工业科技, 2006, 27(9): 185~188.
[20]. J Alberola, A Rodríguez, O Francino, X Roura, L Rivas, and D Andreu. Safety and Efficacy of Antimicrobial Peptides against Naturally Acquired Leishmaniasis. Biopolymers, 1998, 47: 415~433.
[21].卢其能,罗晓明,江辉,邹一平.抗菌肽的研究进展.江西科学, 2007, 25(4): 428~433.
[22]. Vincent du Vigneaud, Charlotte Ressler, John M Swan, Carleton W Roberts, et al. The Synthesis of Oxytocin. J Am Chem Soc, 1954, 76 : 3115~ 3121.
[23]. R B Merrifield. Solid Phase Peptide Synthesis. I. The Synthesis of a Tetrapeptide. J Am Chem Soc, 1963, 85 (14): 2149~2154.
[24].程绍玲,黄文强,何炳林.组合化学中的聚合物研究进展[J].离子交换与吸附, 1999, 15(5): 472~480.
[25]. Heru Chen, Richard K Haynes, Jürgen Scherkenbeck. Synthesis of Cyclichexapeptides using Indirect Cyclization Method [J]. Eur J Org Chem, 2004: 38~47.
[26]. Pinner R W, et al. Trends in infectious diseases mortality in the United States[J]. J Am Med Assoc, 1996, 275, 189~193.
[27]. Prenner E J, McElhaney R N, et al. The interaction of the antibacterial peptide gramicidin S with lipid bilayer model and biological membranes[J]. Biochim Biophys Acta, 1999, 1462, 201~221.
[28]. Tzong-Hsien Lee, Don Rivett, Jerome Werkmeister, Dean Hewish, Marie-Isabel Aguilar. Interaction of amphipathic peptideswith an immobilized model membrane[J]. Letters in Peptide Science, 1999, 6: 371~380.
[29]. PerezPaya E, Houghten R A, Blondelle S E. Functionalized protein-like structures from conformationally defined synthetic combinatorial libraries[J]. J Biol Chem, 1995, 270, 1048~1056.
[30]. PerezPaya E, Houghten R A, Blondelle S E. Determination of the secondary structure of selected melittin analogues with different haemolytic activities[J]. Biochem J, 1994, 299, 587~591.
[31]. Gilon C, Halle D, Chorey M, Selinger Z, Byk G. Backbone cyclization: A New Method for Conferring Conformational Constraint on Peptides[J]. Biopolymers, 1991, 31(6), 745~750.
[1]. V K Sarin, S B H Kent. Quantitative Monitoring of solid phase peptides synthesis by acid-based indicator. Coll Czech Chem Comm, 1988, 53: 2542.
[2]. Fields G B, Fields C G. Solvation effects in solid-phase peptide synthesis. J Am Chem Soc, 1991, 113: 4202~4207.
[3]. George W Anderson, Richard W Young. The Use of Diester Chlorophosphites in Peptide Syntheses. Mixed Anhydrides. J Am Chem Soc, 1952, 74: 5307~5309.
[4]. G W Anderson, F M Callahan. Racemization by the dicyclhexylcarbodiimide method of peptide synthesis. J Am Chem Soc, 1958, 80: 2902~2903.
[5]. F Weygand et al. Gaschromatographische Untersuchung der Racemisierung bei Peptidsynthesen. Angew Chem, 1963, 75 : 282~287.
[6]. Daniel S Kemp, S W Wang, G Busby III, G Hugel. Microanalysis by successive isotopic dilution. New assay for racemic content. J Am Chem Soc, 1970, 92: 1043~ 1055.
[7]. N Izumiya, M Muraoka. Racemization test in peptide synthesis. J Am Chem Soc, 1969, 91: 2391~ 2392.
[8].韩香,顾军.多肽的固相合成[J].天津药学, 2002, 14(1): 7~ 9.
[9]. ShigekiIsomura, PeterWirsching. An Immunotherapeutic Program for the Treatment of Nicotine Addiction: Hapten Design and Synthesis. J Org Chem, 2001, 66: 4115~4121.
[10]. Liren Huang, James C. Quada Jr, et al Design, Synthesis, and Sequence Selective DNA Cleavage of Functional Models of Bleomycin. 1. Hybrids Incorporating a Simple Metal-Complexing Moiety of Bleomycin and Lexitropsin Carriers[J]. Bioconjugate Chem, 1995, 6, 21-33
[11]. Ana Montero, Pilar Goya, Nadine Jagerovic. Guanidinium and Aminoimidazolinium Derivatives of N-(4-Piperidyl)Propanamides as Potential Ligands fouμOpioid and I2-imidazoline Receptors:Synthesis and Pharmacological Screening. Bioorganic and Medicinal Chemistry, 2002, 10: 1009~1018.
[12]. Mira S, Dejan M Godjevac. Synthesis and antioxidant properties of fullero-steroial covalent conjugates. Carbon, 2007, 45: 2260~2265.
[13].王德心.固相有机合成[M].北京:化学工业出版社,2004, 55-58.
[14]. Besser D, Reissmann S, Olender R, Rosenfeld R, Arad O. Study on the cyclizationtendency of backbone cyclic tetrapeptides. J Peptide Res, 2000, 56: 337-345.
[1]. Chen H R, Haynes R K, and Scherkenbeck J. Synthesis of Cyclohexapeptides Based on the Antibiotic Cyclic Decapeptide Loloatin C by an in situ Non-directed Cyclization. European J Org Chem, 2004, 1, 38～47.
[2]. Timothy J Falla, D Nedra Karunaratne and Robert E W Hancock[J]. The Journal of Biological Chemistry. 1996, 273(32): 19298～19303.
[3]. Brenwald N P, Gill M J, Wise R. Prevalence of a putative efflux mechanism among fluoroquinolone resistant clinical isolates[J]. Antimicrob Agents Chemother, 1998, 42(8): 2032～2035.
[4].王玉宝,宋诗铎,魏殿军等.多重耐药泵抑制剂提高肠球菌对氟喹诺酮药物敏感性的研究[J].中国抗生素杂志,2004,29(3):166～168.
[5].魏殿军,宋诗铎,马金玲等.泵抑制剂对耐喹诺酮类铜绿假单胞菌临床株作用的研[J].中国抗生素杂志,2004,29(8):465～468.
[6].陶传敏,李小春,吕晓菊等.氟喹诺酮类药物对嗜麦芽寡养单胞菌的抗菌活性及羰基氰氯苯腙对其的影响[J].中国抗生素杂志,2004,29(8): 469～472.