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HK1175478B - Template-fixed peptidomimetics with antimicrobial activity - Google Patents

Template-fixed peptidomimetics with antimicrobial activity Download PDF

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Publication number
HK1175478B
HK1175478B HK13102561.0A HK13102561A HK1175478B HK 1175478 B HK1175478 B HK 1175478B HK 13102561 A HK13102561 A HK 13102561A HK 1175478 B HK1175478 B HK 1175478B
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HK
Hong Kong
Prior art keywords
dab
lower alkyl
trp
chr
alkyl group
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HK13102561.0A
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Chinese (zh)
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HK1175478A1 (en
Inventor
F．贡贝特
D．奥伯莱希特
W．维利吉伯罗德
R．蒂亚斯
S．J．德马科
J．A．罗宾逊
N．斯里尼瓦斯
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波利弗尔有限公司
苏黎世大学
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Priority claimed from PCT/CH2006/000036 external-priority patent/WO2007079597A1/en
Application filed by 波利弗尔有限公司, 苏黎世大学 filed Critical 波利弗尔有限公司
Publication of HK1175478A1 publication Critical patent/HK1175478A1/en
Publication of HK1175478B publication Critical patent/HK1175478B/en

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Description

Template-fixed peptidomimetics with antimicrobial activity
The application is a divisional application of Chinese patent application with application date of 2007, 1, 15, application number of "200780008052.1" and invention name of "template-fixed peptide mimetic with antimicrobial activity".
Technical Field
The present invention provides template-fixed β -hairpin peptidomimetics incorporating a template-fixed chain of 12 α -amino acid residues which are Gly, or Pro, or a specific type, depending on their position in the chain, as defined below. These template-immobilized beta-hairpin mimetics have selective antimicrobial activity. Furthermore, the present invention provides efficient synthetic methods by which these compounds can be prepared as parallel libraries as desired. These β -hairpin peptidomimetics show improved efficacy, bioavailability, half-life, most importantly a significantly improved ratio between antibacterial activity (on the one hand) and hemolysis of erythrocytes (on the other hand).
Background
The problem of resistance of ever-increasing microorganisms to existing antibiotics has stimulated strong interest in developing new antimicrobial agents with new modes of action (h. breithaupt, nat. biotechnol.1999, 17, 1165-. An emerging class of antibiotics is based on naturally occurring cationic peptides (T.Ganz, R.I.Lehrer, mol.medicine Today 1999, 5, 292-. These include disulfide-bridged β -hairpin and β -sheet peptides (e.g., proteolytic clevidin [ V.N. M.; O.V.Shamova, H.A.Korneva, R.I.Lehrer, FEBSLett.1993, 327, 231. 236.), tachyplesin [ T.Nakamura, H.Furunaka, T.Miyata, F.Tokunaga, T.Muta, S.Iwana, M.Niwa, T.Takao, Y.Shimonishi, Y.J.biol.Chem.1988, 263, 16709. 16713], and defensins [ R.I.Lehrer, A.K.Lichtenstein, T.Ganz, Annu.Rev.Imol.1993, 11, 105. 128], amphipathic α -helical peptides (e.g., amphipathic peptides, sanchenginine, and β -pleated peptides [ 7. sane.S.S.I.S.I.Lehrer., T.S.I.I.S.S.S.S.S.S.Niwa, T.Takao, Y.S.S.S.S.S.S.Shimoni., Y., T.S.S.S. Shimoni., 11, 105, E.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.S.A.S.S.S.S.S.A.S.A.A.A.S.A, more complex mechanisms of action, e.g. involving receptor mediated signalling, cannot be ruled out at present (M.Wu, E.Maier, R.Benz, R.E.Hancock, Biochemistry1999, 38, 7235-.
The antimicrobial activity of many of these cationic peptides is often associated with their preferred secondary structure, whether observed in aqueous solution or in a membrane-like environment (n.sitaram, r.nagaraj, biochim.biophysis.acta1999, 1462, 29-54). Structural studies by Nuclear Magnetic Resonance (NMR) spectroscopy have shown that cationic peptides such as, for example, proteinase 1 (a. aumelas, m. mangoni, c. roumesland, l. chiche, e.despaux, g.grassy, b.calas, a.chananieu, a.eur.j.biochem.1996, 237, 575-583; r.l.fahrner, t.dieckmann, s.s.l.harwig, r.i.lehrer, d.eisenberg, j.feigon, j.chem.biol.1996, 3, 543-550) and limulus I (k.kawano, t.yoneya, t.miyata, k.yoshikawa, f.tokunaga, y.terada, s.j.iwannaa, s.iwanna, s.15365, 1990. beta. disulfide bridges appear to be restricted by the two hairpin conformations. In a lysin analogue lacking one or both of these disulfide bonds, the stability of the β -hairpin conformation is reduced and the antimicrobial activity is reduced (J.Chen, T.J.Falla, H.J.Liu, M.A.Hurst, C.A.Fujii, D.A.Mosca, J.R.Embree D.J.Loury, P.A.Radel, C.C.Chang, L.Gu, J.C.Fiddes, Biopolymers2000, 55, 88-98; S.L.Harwing, A.Waring, H.J.Yang, Y.Cho, L.Tan, R.I.Lehrer, R.J.Eur.J.J.J.1996, 240, 352- -357, M.E.goniMan, A.A.Memelas, P.Aurentz, C.Charnaesters, Rough.L.J.Walsh, J.J.J.J.Eur.J.J.J.J.J.J.J.1996, 240, 352-. Similar observations were made for analogs of tachyplesin I (h.tamamamura, r.ikoma, m.niwa, s.funakoshi, t.murakami, n.fujii, chem.pharm.bull.1993, 41, 978-plus 980) and hairpin-loop mimetics of rabbit defensin NP-2 (s.thermarasu, r.nagaraj, biochem.biophysis.res.comm.1999, 254, 281-plus 283). These results indicate that the β -hairpin structure plays an important role in the antimicrobial activity and stability of these proteolysin-like peptides. In the case of cationic peptides that favor the alpha-helical structure, the amphipathic structure of the helix appears to play a key role in determining antimicrobial activity (a.tossi, l.sandri, a.gianga sporo, a.biopolymers2000, 55, 4-30). Gramicidin S is a backbone-cyclic peptide with a well-defined β -hairpin structure (s.e.hull, r.karlsson, p.main, m.m.woolfson, e.j.dodson, Nature1978, 275, 206-. However, the high hemolytic activity of gramicidin S prevents its widespread use as an antibiotic. Recent structural studies by NMR show that high hemolytic activity is clearly related to the highly amphipathic nature of the cyclic β -hairpin-like molecule, but that antimicrobial activity and hemolytic activity can be separated by adjusting the conformation and amphiphilicity (l.h.kondejewski, m.jelokhani-Niaraki, s.w.farmer, b.lix, m.kay, b.d.sykes, r.e.cohanck, r.s.hodes, j.biol.chem.1999, 274, 13181-jar 13192; c.mcinnes l.h.kondejewski, r.s.hodges, b.d.sys, j.biol.chem.2000, 275, 14287-jar 94).
A novel cyclic antimicrobial peptide RTD-1 (y. -q. tang, j. yuan, G) from primate white blood cells has recently been reported.K.Tran, c.j.miller, a.j.oelette, m.e.selsted, Science1999, 286, 498-. This peptide contains three disulfide bridges which serve to constrain the cyclic peptide backbone to a hairpin geometry. The cleavage of these three disulfide bonds results in a significant loss of antimicrobial activity. Also reported are analogs of lysins (J.P.Tam, C.Wu, J. -L.Yang, Eur.J.Biochem.2000, 267, 3289-. In these cases, all cystine-limited removal did not always result in a large loss of antimicrobial activity, but the membrane-lysing selectivity was adjusted (J.P.Tam, C.Wu, J. -L.Yang, Eur.J.biochem.2000, 267, 3289-.
Key issues in the design of new selective cationic antimicrobial peptides are bioavailability, stability and reduced hemolytic activity. Naturally occurring proteolytic enzymes and tachyplesin produce significant hemolytic activity against human erythrocytes. This is also the case for a proteoliposome analogue such as IB367 (J.Chen, T.J.Falla, H.J.Liu, M.A.Hurst, C.A.Fujii, D.A.Mosca, J.R.Embree, D.J.Loury, P.A.Radel, C.C.Chang, L.Gu, J.C.Fiddes, Biopolymers2000, 55, 88-98; C.Chang, L.Gu, J.Chen, U.S. Pat: 5,916,872, 1999). This high hemolytic activity substantially excludes its use in vivo and represents a serious drawback in clinical applications. In addition, the antibiotic activity of the analogs generally decreased significantly with increasing salt concentration, resulting in a significant decrease in antimicrobial activity under in vivo conditions (about 100-.
The proteolysin 1 exhibited efficacy and similar activity in low and high salt assays against gram positive and gram negative bacteria as well as fungi. This broad antimicrobial activity coupled with a rapid mode of action, and their ability to kill bacteria resistant to other classes of antibiotics, makes them attractive targets for the development of clinically useful antibiotics. Activity against gram-positive bacteria is generally higher than against gram-negative bacteria. However, proteinase 1 also exhibits high hemolytic activity against human erythrocytes and thus low selectivity for microbial cells. Targeted CD experiments (W.T.Heller, A.J.Waring, R.I.Lehrer, H.W.Huang, Biochemistry 1998, 37, 17331-. Studies on cyclic lysin analogues (J. -P.Tam, C.Wu, J. -L.Yang, Eur.J.biochem.2000, 267, 3289-Buchner 3300) have shown that an increase in conformational rigidity caused by backbone cyclisation and multiple disulphide bridges can produce lysomembrane selectivity, which separates antimicrobial activity from hemolytic activity, at least in the series of compounds studied.
Proteinase 1 is an 18-residue linear peptide with an amidated carboxy terminus and two disulfide bridges. Limulus I contains 17 residues, also has an amidated carboxy terminus and contains two disulfide bridges. Recently described backbone-cyclic lysins and limulus analogs typically contain 18 residues and up to three disulfide bridges (J.P.Tam, C.Wu, J.L.Yang, Eur.J.biochem.2000, 267, 3289-.
Antibacterial peptides (cathelicidins) (37-residue linear helical cationic peptides) and analogues currently under investigation are used as inhalation therapeutics for Cystic Fibrosis (CF) lung disease (L.Saiman, S.Tabibi, T.D.Starner, P.san Gabriel, P.L.Winokur, H.P.Jia, P.B.McGray, Jr., B.F.Tack, Antimicrob.AgentandChemothert.2001, 45, 2838-2844; R.E.W.Hancock, R.Lehrer, trends Biotechnol.1998, 16, 82-88). Over 80% of CF patients suffer chronic infection with P.aeruginosa (C.A.Demko, P.J.Biard, P.B.Davies, J.Clin.epidemiol.1995, 48, 1041-. Other antimicrobial peptides against pseudomonas (y.h. yau, b.ho, n.s.tan, m.l.ng, j.l.ding, antimicrob.agentan and chemicother.2001, 45, 2820-2825 and references cited therein), such as FALL-39, SMAP-29, and lepidopteran insecticidal peptides, exhibit less desirable properties, such as effective antimicrobial activity over a wide range of pH, rapid kill rates, and low hemolytic activity.
Among the compounds described below, a novel strategy was introduced to stabilize the β -hairpin conformation in backbone-cyclic cationic peptidomimetics that exhibit selective antimicrobial activity. This involves grafting cationic and hydrophobic hairpin sequences onto a template that functions to constrain the peptide loop backbone to a hairpin geometry.
Template-binding hairpin mimetic peptides (D, Obrecht, M.Altorfer, J.A.Robinson, adv.Med.Chem.1999, 4, 1-68; J.A.Robinson, Syn.Lett.2000, 4, 429-441) have been described in the literature, and the ability to generate β -hairpin peptidomimetics using combinatorial and parallel synthesis methods has been established (L.Jiang, K.Moehle, B.Dhanapaal, D.Obrecht, J.A.Robinson, Helv.Chim.acta.2000, 83, 3097-3113112). Antibacterial template-fixed peptidomimetics and methods for their synthesis have been described in international patent applications WO02/070547A1 and WO2004/018503A1, but these molecules do not exhibit high selectivity of plasma stability and particularly high efficacy.
Disclosure of Invention
The methods described herein enable the synthesis and screening of large pools of hairpin mimetics, which subsequently contributes significantly to structure-activity studies, and thus the discovery of novel molecules with potent selective antimicrobial activity and very low hemolytic activity on human erythrocytes. The method enables the synthesis of beta-hairpin peptidomimetics with novel selectivity against various multidrug resistant pseudomonas strains.
The beta-hairpin peptidomimetics of the invention are compounds having the general formula
Wherein
Is a radical of one of the formulae
Wherein
Is a residue of an L-alpha-amino acid, and B is of the formula-NR20CH(R71) -or an enantiomer of one of the groups a1 to a69 as defined below;
is a radical of one of the formulae
R1Is H; a lower alkyl group; or aryl-lower alkyl;
R2is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R3Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R4Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)p(CHR61)sCOOR57；-(CH2)p(CHR61)sCONR58R59；-(CH2)p(CHR61)sPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R5Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R6Is H; an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R7Is an alkyl group; an alkenyl group; - (CH)2)q(CHR61)sOR55；-(CH2)q(CHR61)sNR33R34；-(CH2)q(CHR61)sOCONR33R75；-(CH2)q(CHR61)sNR20CONR33R82；-(CH2)r(CHR61)sCOOR57；-(CH2)r(CHR61)sCONR58R59；-(CH2)r(CHR61)sPO(OR60)2；-(CH2)r(CHR61)sSO2R62(ii) a Or- (CH)2)r(CHR61)sC6H4R8；
R8Is H; cl; f; CF (compact flash)3；NO2(ii) a A lower alkyl group; lower alkenyl; an aryl group; aryl-lower alkyl; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)NR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sCOR64；
R9Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R10Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R11Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R12Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)r(CHR61)sCOOR57；-(CH2)r(CHR61)sCONR58R59；-(CH2)r(CHR61)sPO(OR60)2；-(CH2)r(CHR61)sSO2R62(ii) a Or- (CH)2)r(CHR61)sC6H4R8；
R13Is an alkyl group; an alkenyl group; - (CH)2)q(CHR61)sOR55；-(CH2)q(CHR61)sSR56；-(CH2)q(CHR61)sNR33R34；-(CH2)q(CHR61)sOCONR33R75；-(CH2)q(CHR61)sNR20CONR33R82；-(CH2)q(CHR61)sCOOR57；-(CH2)q(CHR61)sCONR58R59；-(CH2)q(CHR61)sPO(OR60)2；-(CH2)q(CHR61)sSO2R62(ii) a Or- (CH)2)q(CHR61)sC6H4R8；
R14Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)q(CHR61)sCOOR57；-(CH2)q(CHR61)sCONR58R59；-(CH2)q(CHR61)sPO(OR60)2；-(CH2)q(CHR61)sSOR62(ii) a Or- (CH)2)q(CHR61)sC6H4R8；
R15Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R16Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R17Is an alkyl group; an alkenyl group; - (CH)2)q(CHR61)sOR55；-(CH2)q(CHR61)sSR56；-(CH2)q(CHR61)sNR33R34；-(CH2)q(CHR61)sOCONR33R75；-(CH2)q(CHR61)sNR20CONR33R82；-(CH2)q(CHR61)sCOOR57；-(CH2)q(CHR61)sCONR58R59；-(CH2)q(CHR61)sPO(OR60)2；-(CH2)q(CHR61)sSO2R62(ii) a Or- (CH)2)q(CHR61)sC6H4R8；
R18Is an alkyl group; an alkenyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sSR56；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)p(CHR61)sCOOR57；-(CH2)p(CHR61)sCONR58R59；-(CH2)p(CHR61)sPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R19Is a lower alkyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sSR56；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)p(CHR61)sCOOR57；-(CH2)p(CHR61)sCONR58R59；-(CH2)p(CHR61)sPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8(ii) a Or
R18And R19Together may form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；
R20Is H; an alkyl group; an alkenyl group; or aryl-lower alkyl;
R21is H; an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R22Is H; an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R23Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR6)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R24Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R25Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R26Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8(ii) a Or
R25And R26Together may form: - (CH)2)2-6-；-(CXH)rO(CH2)r-；-(CH2)rS(CH2)r-; or- (CH)2)rNR57(CH2)r-；
R27Is H; an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R28Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)s-OR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R29Is an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R30Is H; an alkyl group; an alkenyl group; or aryl-lower alkyl;
R31is H; an alkyl group; an alkenyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R32Is H; a lower alkyl group; or aryl-lower alkyl;
R33is H; alkyl, alkenyl; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR34R63；-(CH2)m(CHR61)sOCONR75R82；-(CH2)m(CHR61)sNR20CONR78R82；-(CH2)o(CHR61)sCOR64；-(CH2)o(CHR61)s-CONR58R59，-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R34Is H; a lower alkyl group; an aryl group, a heteroaryl group,or aryl-lower alkyl;
R33and R34Together may form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；
R35Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)p(CHR61)sCOOR57；-(CH2)p(CHR61)sCONR58R59；-(CH2)p(CHR61)sPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)p(CHR61)sC6H4R8；
R36Is H, alkyl; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)p(CHR61)sCOOR57，-(CH2)p(CHR61)sCONR58R59；-(CH2)p(CHR61)sPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R37Is H; f; br; cl; NO2；CF3(ii) a A lower alkyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R38Is H; f; br; cl; NO2；CF3(ii) a An alkyl group; an alkenyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R39Is H; an alkyl group; an alkenyl group; or aryl-lower alkyl;
R40is H; an alkyl group; an alkenyl group; or aryl-lower alkyl;
R41is H; f; br; cl; NO2；CF3(ii) a An alkyl group; an alkenyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R42Is H; f; br; cl; NO2；CF3(ii) a An alkyl group; an alkenyl group; - (CH)2)p(CHR61)sOR55；-(CH2)p(CHR61)sNR33R34；-(CH2)p(CHR61)sOCONR33R75；-(CH2)p(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R43Is H, alkyl; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57，-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)sPO(OR60)2；-(CH2)o(CHR61)sSO2R62(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R44Is an alkyl group; an alkenyl group; - (CH)2)r(CHR61)sOR55；-(CH2)r(CHR61)sSR56；-(CH2)r(CHR61)sNR33R34；-(CH2)r(CHR61)sOCONR33R75；-(CH2)r(CHR61)sNR20CONR33R82；-(CH2)r(CHR61)sCOOR57；-(CH2)r(CHR61)sCONR58R59；-(CH2)r(CHR61)sPO(OR60)2；-(CH2)r(CHR61)sSO2R62(ii) a Or- (CH)2)r(CHR61)sC6H4R8；
R45Is H; an alkyl group; an alkenyl group; - (CH)2)o(CHR61)sOR55；-(CH2)o(CHR61)sSR56；-(CH2)o(CHR61)sNR33R34；-(CH2)o(CHR61)sOCONR33R75；-(CH2)o(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)s(CHR61)sCONR58R59；-(CH2)s(CHR61)sPO(OR60)2；-(CH2)s(CHR61)sSO2R62(ii) a Or- (CH)2)s(CHR61)sC6H4R8；
R46Is H; an alkyl group; an alkenyl group; or- (CH)2)o(CHR61)pC6H4R8；
R47Is H; an alkyl group; an alkenyl group; or- (CH)2)o(CHR61)sOR55；
R48Is H; a lower alkyl group; lower alkenyl; or aryl-lower alkyl;
R49is H; an alkyl group; an alkenyl group; - (CHR)61)sCOOR57；(CHR61)sCONR58R59；(CHR61)sPO(OR60)2；-(CHR61)sSOR62(ii) a Or- (CHR)61)sC6H4R8；
R50Is H; a lower alkyl group; or aryl-lower alkyl;
R51is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)pPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)p(CHR61)sC6H4R8；
R52Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)pPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)p(CHR61)sC6H4R8；
R53Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sSR56；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)sCOOR57；-(CH2)o(CHR61)sCONR58R59；-(CH2)o(CHR61)pPO(OR60)2；-(CH2)p(CHR61)sSO2R62(ii) a Or- (CH)2)p(CHR61)sC6H4R8；
R54Is H; an alkyl group; an alkenyl group; - (CH)2)m(CHR61)sOR55；-(CH2)m(CHR61)sNR33R34；-(CH2)m(CHR61)sOCONR33R75；-(CH2)m(CHR61)sNR20CONR33R82；-(CH2)o(CHR61)COOR57；-(CH2)o(CHR61)sCONR58R59(ii) a Or- (CH)2)o(CHR61)sC6H4R8；
R55Is H; a lower alkyl group; lower alkenyl; aryl-lower alkyl; - (CH)2)m(CHR61)sOR57；-(CH2)m(CHR61)sNR34R63；-(CH2)m(CHR61)sOCONR75R82；-(CH2)m(CHR61)sNR20CONR78R82；-(CH2)o(CHR61)s-COR64；-(CH2)o(CHR61)COOR57(ii) a Or- (CH)2)o(CHR61)sCONR58R59；
R56Is H; a lower alkyl group; lower alkenyl; aryl-lower alkyl; - (CH)2)m(CHR61)sOR57；-(CH2)m(CHR61)sNR34R63；-(CH2)m(CHR61)sOCONR75R82；-(CH2)m(CHR61)sNR20CONR78R82；-(CH2)o(CHR61)s-COR64(ii) a Or- (CH)2)o(CHR61)sCONR58R59；
R57Is H; a lower alkyl group; lower alkenyl; aryl lower alkyl; or heteroaryl lower alkyl;
R58is H; a lower alkyl group; lower alkenyl; an aryl group; a heteroaryl group; aryl-lower alkyl; or heteroaryl-lower alkyl;
R59is H; a lower alkyl group; lower alkenyl; an aryl group; a heteroaryl group; aryl-lower alkyl; or heteroaryl-lower alkyl; or
R58And R59Together may form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；
R60Is H; a lower alkyl group; lower alkenyl; an aryl group; or aryl-lower alkyl;
R61is H; an alkyl group; an alkenyl group; an aryl group; a heteroaryl group; aryl-lower alkyl; heteroaryl-lower alkyl; - (CH)2)pOR55；-(CH2)pNR33R34；-(CH2)pOCONR75R82；-(CH2)pNR20CONR78R82；-(CH2)oCOOR37(ii) a Or- (CH)2)oPO(COR60)2；
R62Is a lower alkyl group; lower alkenyl; aryl, heteroaryl; or aryl-lower alkyl;
R63is H; a lower alkyl group; lower alkenyl; aryl, heteroaryl; aryl-lower alkyl; heteroaryl-lower alkyl; -COR64；-COOR57；-CONR58R59；-SO2R62(ii) a OR-PO (OR)60)2；
R34And R63Together may be formed; - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；
R64Is H; a lower alkyl group; lower alkenyl; an aryl group; a heteroaryl group; aryl-lower alkyl; heteroaryl-lower alkyl; - (CH)2)p(CHR61)sOR65；-(CH2)p(CHR61)sSR66(ii) a Or- (CH)2)p(CHR61)sNR34R63；-(CH2)P(CHR61)sOCONR75R82；-(CH2)P(CHR61)sNR20CONR78R82；
R65Is H; a lower alkyl group; lower alkenyl; aryl, aryl-lower alkyl; heteroaryl-lower alkyl; -COR57；-COOR57(ii) a or-CONR58R59；
R66Is H; a lower alkyl group; lower alkenyl; an aryl group; aryl-lower alkyl; heteroaryl-lower alkyl; or-CONR58R59；
m is 2 to 4; o is 0 to 4; p is 1 to 4; q is 0 to 2; r is 1 or 2; s is 0 or 1;
R67is H; cl; br; f; NO2；-NR34COR57(ii) a A lower alkyl group; or lower alkenyl;
R68is H; cl; br; f; NO2；-NR34COR57(ii) a A lower alkyl group; or lower alkenyl;
R69is H; cl; br; f; NO2；-NR34COR57(ii) a A lower alkyl group; or lower alkenyl; and
R70is H; cl; br; f; NO2；-NR34COR57(ii) a A lower alkyl group; or lower alkenyl;
with the proviso that R67、R68、R69And R70At least two of (a) are H; and
z is a chain of 12 alpha-amino acid residues, the positions of said amino acid residues in said chain being counted starting from the N-terminal amino acid, whereby these amino acid residues are Gly or Pro, or of the formula-A-CO-, or of the formula-B-CO-, or of one of the following types, depending on their position in the chain
C：-NR20CH(R72)CO-；
D：-NR20CH(R73)CO-；
E：-NR20CH(R74)CO-；
F：-NR20CH(R84)CO-；
H：-NR20-CH(CO-)-(CH2)4-7-CH(CO-)-NR20-；
-NR20-CH(CO-)-(CH2)pSS(CH2)p-CH(CO-)-NR20-；
-NR20-CH(CO-)-(-(CH2)pNR20CO(CH2)p-CH(CO-)-NR20-; and
-NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-；
R71is H; a lower alkyl group; lower alkenyl; - (CX)2)p(CHR61)sOR75；-(CH2)p(CHR61)sSR75；-(CX2)p(CHR61)sNR33R34；-(CX2)p(CHR61)sOCONR33R75；-(CX2)p(CHR61)sNR20CONR33R82；-(CX2)o(CHR61)sCOOR75；-(CX2)pCONR58R59；-(CX2)pPO(OR62)2；-(CX2)pSO2R62(ii) a Or- (CX)2)o-C6R67R68R69R70R76；
R72Is H; a lower alkyl group; lower alkenyl; - (CX)2)p(CHR86)sOR85(ii) a Or- (CX)2)p(CHR86)sSR85；
R73Is- (CX)2)oR77；-(CX2)rO(CH2)oR77；-(CX2)rS(CH2)oR77；
Or- (CX)2)rNR20(CH2)oR77；
R74Is- (CX)2)pNR78R79；-(CX2)pNR77R80；-(CX2)pC(=NR80)NR78R79；-(CX2)pC(=NOR50)NR78R79；-(CX2)pC(=NNR78R79)NR78R79；-(CX2)pNR80C(=NR80)NR78R79；-(CX2)pN=C(NR78R80)NR79R80；-(CX2)pC6H4NR78R79；-(CX2)pC6H4NR77R80；-(CX2)pC6H4C(=NR80)NR78R79；-(CX2)pC6H4C(=NOR50)NR78R79；-(CX2)pC6H4C(=NNR78R79)NR78R79；-(CX2)pC6H4NR80C(=NR80)NR78R79；-(CX2)pC6H4N=C(NR78R80)NR79R80；-(CX2)rO(CX2)mNR78R79；-(CX2)rO(CX2)mNR77R80；-(CX2)rO(CX2)pC(=NR80)NR78R79；-(CX2)rO(CX2)pC(=NOR50)NR78R79；-(CX2)rO(CX2)pC(=NNR78R79)NR78R79；-(CX2)rO(CH2)mNR80C(=NR80)NR78R79；-(CX2)rO(CX2)mN=C(NR78R80)NR79R80；-(CX2)rO(CX2)pC6H4CNR78R79；-(CX2)rO(CX2)pC6H4C(=NR80)NR78R79；-(CX2)rO(CX2)pC6H4C(=NOR50)NR78R79；-(CX2)rO(CX2)pC6H4C(=NNR78R79)NR78R79；-(CX2)rO(CX2)pC6H4NR80C(=NR80)NR78R79；-(CX2)rS(CX2)mNR78R79；-(CX2)rS(CX2)mNR77R80；-(CX2)rS(CX2)pC(=NR80)NR78R79；-(CX2)rS(CX2)pC(=NOR50)NR78SR79；-(CX2)rS(CX2)pC(=NNR78R79)NR78R79；-(CX2)rS(CX2)mNR80C(=NR80)NR78R79；-(CX2)rS(CX2)mN=C(NR78R80)NR79R80；-(CX2)rS(CX2)pC6H4CNR78R79；-(CX2)rS(CX2)pC6H4C(=NR80)NR78R79；-(CX2)rS(CX2)pC6H4C(=NOR50)NR78R79；-(CX2)rS(CX2)pC6H4C(=NNR78R79)NR78R79；-(CX2)rS(CX2)pC6H4NR80C(=NR80)NR78R79；-(CX2)pNR80COR64；-(CX2)pNR80COR77；-(CX2)pNR80CONR78R79；-(CX2)pC6H4NR80CONR78R79(ii) a Or- (CX)2)pNR20CO-[(CX2)u-XX]t-CH3Wherein XX is-O-; -NR20-, or-S-; u is 1 to 3 and t is 1 to 6.
R75Is a lower alkyl group; lower alkenyl; or aryl-lower alkyl;
R33and R75Together may form: - (CX)2)2-6-；-(CX2)2O(CH2)2-；-(CX2)2S(CX2)2-; or- (CX)2)2NR57(CX2)2-；
R75And R82Together may form: - (CX)2)2-6-；-(CX2)2O(CX2)2-；-(CX2)2S(CX2)2-; or- (CX)2)2NR57(CX2)2-；
R76Is H; lower alkyl(ii) a Lower alkenyl; aryl-lower alkyl; - (CX)2)oOR72；-(CX2)oSR72；-(CX2)oNR33R34；-(CX2)oOCONR33R75；-(CX2)oNR20CONR33R82；-(CX2)oCOOR75；-(CX2)oCONR58R59；-(CX2)oPO(OR60)2；-(CX2)pSO2R62(ii) a Or- (CX)2)oCOR64；
R77is-C6R67R68R69R70R76(ii) a Or a heteroaryl group having one of the following general formulae
R78Is H; a lower alkyl group; an aryl group; or aryl-lower alkyl;
R78and R82Together may form: - (CX)2)2-6-；-(CX2)2O(CX2)2-；-(CX2)2S(CX2)2-; or- (CX)2)2NR57(CX2)2-；
R79Is H; a lower alkyl group; an aryl group; or aryl-lower alkyl; or
R78And R79Taken together, may be- (CX)2)2-7-；-(CX2)2O(CX2)2-; or- (CX)2)2NR57(CX2)2-；
R80Is H; or lower alkyl;
R81is H; a lower alkyl group; or aryl-lower alkyl;
R82is H; a lower alkyl group; an aryl group; a heteroaryl group; or aryl-lower alkyl;
R33and R82Together may form: - (CX)2)2-6-；-(CX2)2O(CX2)2-；-(CX2)2S(CX2)2-; or- (CX)2)2NR57(CX2)2-；
R83Is H; a lower alkyl group; an aryl group; or-NR78R79；
R84Is- (CX)2)m(CHR61)sOH；-(CX2)pCONR78R79；-(CX2)pNR80CONR78R79；-(CX2)pC6H4CONR78R79(ii) a Or- (CX)2)pC6H4NR80CONR78R79；
R85Is a lower alkyl group; or lower alkenyl;
R86is H; an alkyl group; an alkenyl group; or- (CX)2)pOR85；-(CX2)pSR85
R87Is H; an alkyl group; an alkenyl group; a heteroaryl group; aryl-lower alkyl; - (CX)2)pOR55；-(CX2)pOCONR75R82；-(CX2)pNR20CONR78R82；-(CX2)pCOOR57Or is or-(CX2)pPO(OR60)2；
X is H, or optionally halogen;
with the proviso that in a preferred embodiment, in the chain of 12 α -amino acid residues Z, the amino acid residues in positions 1 to 12 are:
-P1: type C or type D or type E or type F, or the residue is Pro;
-P2: type D or type E;
-P3: type C, or type D, or residue is Gly or Pro;
-P4: type C, or type E, or type F; or residue is Gly or Pro;
-P5: type E, or type D, or type C, or residue is Gly or Pro;
-P6: type E, or type F or type C, or of the general formula-A-CO-, or the residue is Gly or Pro;
-P7: type C, or type E or type F or of the general formula-B-CO-;
-P8: type D, or type C, or type F, or the residue is Pro;
-P9: type C, or type E, or type D, or type F;
-P10: type E;
-P11: type C or type F; or residue is Pro or Gly; and
-P12: type C or type D or type E or type F, or the residue is Pro; or
-P4 and P9 and/or P2 and P11 together may form a group of type H; and may also be the D-isomer at P6, P10 and P11;
or, alternatively, but in less preferred embodiments:
-P1: type C or type D or type E or type F, or the residue is Pro;
-P2: type C or type F, or the residue is Pro or Gly;
-P3: type E;
-P4: type C, or type E, or type D or type F;
-P5: type D, or type C, or type F, or the residue is Pro;
-P6: type C, or type E or type F, of the general formula-B-CO-;
-P7: type E, or type F or type C or of the formula-A-CO-, or the residue is Gly or Pro;
-P8: type E, or type D, or type C, or residue is Gly or Pro;
-P9: type C, or type E or type F; or residue is Gly or Pro;
-P10: type C or type D, or residue is Gly or Pro;
-P11: type D or type E; and
-P12: type C or type D or type E or type F, or the residue is Pro; or
-P4 and P9 and/or P2 and P11 together may form a group of type H; and may also be the D-isomer at P2, P3 and P7;
and pharmaceutically acceptable salts thereof.
According to the invention, these β -hairpin peptidomimetics can be prepared by a method comprising:
(a) coupling an appropriately functionalized solid support with an appropriate N-protected derivative of an amino acid at position 5, 6 or 7 in the desired end product, any functional groups that may be present in said N-protected amino acid derivative likewise being appropriately protected;
(b) removing the N-protecting group from the product thus obtained;
(c) coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position in the desired end product close to the N-terminal amino acid residue, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(d) removing the N-protecting group from the product thus obtained;
(e) repeating steps (c) and (d) until the N-terminal amino acid residue has been introduced;
(f) coupling the product thus obtained with a compound of the general formula
Wherein
As defined above and X is an N-protecting group, or, if
The above group (a1) or (a2), as an alternative,
(fa) coupling the product obtained in step (e) with a suitable N-protected derivative of an amino acid having the general formula: HOOC-B-H III or HOOC-A-H IV
Wherein B and A are as defined above, any functional groups which may be present in the N-protected amino acid derivative are likewise suitably protected;
(fb) removing the N-protecting group from the product thus obtained; and
(fc) coupling of the product thus obtained with a suitable N-protected derivative of an amino acid having the general formulae IV and III above, respectively, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(g) removing the N-protecting group from the product obtained in step (f) or (fc);
(h) coupling the product thus obtained with a suitable N-protected derivative of the amino acid at position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative likewise being suitably protected;
(i) removing the N-protecting group from the product thus obtained;
(j) coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position remote from position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(k) removing the N-protecting group from the product thus obtained;
(l) Repeating steps (j) and (k) until all amino acid residues have been introduced;
(m) if desired, selectively deprotecting one or several of the protected functional groups present in the molecule and appropriately substituting the reactive groups thus released;
(o) separating the product thus obtained from the solid support;
(p) cyclizing the product isolated from the solid support;
(q) if desired, forming one or two interchain linkages between the side chains of suitable amino acid residues at opposite positions of the β -strand segment;
(r) removing any protecting groups present on the functional groups of any members of the chain of amino acid residues and, if desired, any protecting groups that may additionally be present in the molecule; and
(s) if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable or unacceptable salt thus obtained into the corresponding free compound of the general formula I or into a different pharmaceutically acceptable salt.
Alternatively, peptidomimetics of the invention can be prepared as follows:
(a') coupling an appropriately functionalized solid support with a compound of the general formula
Wherein
As defined above and X is an N-protecting group, or, if
The above group (a1) or (a2) as an alternative
(a' a) coupling said suitably functionalized solid support with a suitable N-protected derivative of an amino acid having the general formula: HOOC-B-H III or HOOC-A-H IV
Wherein B and A are as defined above, any functional groups which may be present in the N-protected amino acid derivative are likewise suitably protected;
(a' b) removing the N-protecting group from the product thus obtained; and
(a' c) coupling the product thus obtained with a suitable N-protected derivative of an amino acid having the general formulae IV and III above, respectively, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(b ') removing the N-protecting group from the product obtained in step (a ') or (a ' c);
(c') coupling the product thus obtained with a suitable N-protected derivative of the amino acid at position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative likewise being suitably protected;
(d') removing the N-protecting group from the product thus obtained;
(e') coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position remote from position 12 in the desired end product, any functional group which may be present in said N-protected amino acid derivative being likewise suitably protected;
(f') removing the N-protecting group from the product thus obtained;
(g ') repeating steps (e ') and (f ') until all amino acid residues have been introduced;
(h') if desired, selectively deprotecting one or several of the protected functional groups present in the molecule and appropriately substituting the reactive groups thus released;
(i') separating the product thus obtained from the solid support;
(j') cyclizing the product isolated from the solid support;
(k') if desired, forming one or two interchain bonds between the side chains of the appropriate amino acid residues at opposite positions in the β -chain region;
(l') removing any protecting groups present on the functional groups of any members of the chain of amino acid residues and, if desired, any protecting groups which may additionally be present in the molecule; and
(m') if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable or unacceptable salt thus obtained into the corresponding free compound of the general formula I or into a different pharmaceutically acceptable salt.
The peptidomimetics of the invention may also be enantiomers of compounds having the general formula I. These enantiomers can be prepared by varying the above process, wherein all enantiomers of chiral starting materials are used.
The term "alkyl" as used in the present specification, alone or in combination, denotes a saturated straight or branched chain hydrocarbon radical having up to 24, preferably up to 12 carbon atoms, optionally substituted by halogen. Similarly, the term "alkenyl" denotes a straight or branched chain hydrocarbon group having up to 24, preferably up to 12 carbon atoms and containing at least one or, depending on the chain length, up to four olefinic double bonds, optionally substituted by halogen. The term "lower" denotes groups and compounds having up to 6 carbon atoms. Thus, for example, the term "lower alkyl" denotes a saturated straight or branched chain hydrocarbon group having up to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl and the like. The term "aryl" denotes an aromatic carbocyclic hydrocarbon radical comprising one or two six-membered rings, such as phenyl or naphthyl, which may be substituted by up to three substituents such as Br, Cl, F, CF3，NO2Lower alkyl or lower alkenyl. The term "heteroaryl" denotes an aromatic heterocyclic group comprising one or two five-and/or six-membered rings, wherein at least one ring contains up to three heteroatoms selected from O, S and N and the ring is optionally substituted; representative examples of such optionally substituted heteroaryl groups are defined above for R77Is given in the following.
The structural component-A-CO-represents an amino acid structural unit, which binds to the structural components-B-CO-to form the templates (a1) and (a 2). Templates (a) to (p) constitute building blocks having an N-terminus and a C-terminus, which are spatially oriented such that the distance between these two groups may be 4.0-5.5A. The peptide chain Z is linked to the C-and N-termini of the templates (a) to (p) via the respective N-and C-termini, such that the templates and the chain form a cyclic structure, as described in general formula I. In the case where the distance between the N-and C-termini of the template is 4.0-5.5A, the template will induce the H-bond network required for the formation of the beta-hairpin conformation in peptide chain Z. Thus, the template and peptide chain form a β -hairpin mimetic.
The beta-hairpin conformation is highly relevant for the antibacterial activity of the beta-hairpin mimetics of the invention. The β -hairpin stabilizing conformational properties of the templates (a) to (p) play a crucial role not only for selective antibacterial activity but also for the synthesis process as defined above, since the introduction of the template at or near the beginning or the middle of the linearly protected peptide precursor significantly improves the cyclization reaction yield.
The structural units A1-A69 belong to the class of amino acids in which the N-terminus is the secondary amine-forming part of the ring. Among the amino acids encoded by the gene, only proline falls into this class. The structural units A1 to A69 are in configuration (D) and they are bound together with the structural unit-B-CO-in configuration (L). The preferred combination of templates (a1) is-DA1-CO-LB-CO-di-carboxylic acidDA69-CO-LB-CO-. Thus, for example,DPro-Lpro constitutes a prototype of template (a 1). Less preferred but possibleLA1-CO-DB-CO-andLA69-CO-DB-CO-shaping template (a 2). Thus, for example,LPro-Dpro constitutes a prototype of template (a 2).
It is understood that the structural unit in which A has the (D) -configuration-A1-CO-to-A69-CO-carries the group R in the alpha-position to the N-terminus1。R1Are H and lower alkyl, most preferred R1The values are H and methyl. It will be appreciated by those skilled in the art that A1-A69 are shown in the (D) -configuration for R1H and methyl correspond to the (R) -configuration. Depending on the rule according to Cahn, Ingold and Prelog-ruleThen R1Other values of preference, this configuration must also be denoted (S).
Except that R1The structural units-A1-CO-to-A69-CO-may carry further units called R2-R17A substituent of (1). The other substituent may be H, and if it is not H, it is preferably a small to medium size aliphatic or aromatic group. R2To R17Examples of preferred values of (d) are:
-R2: h; a lower alkyl group; lower alkenyl; (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; r33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；R57: h; or lower alkyl); (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R3: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; a lower alkyl group; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R4: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R5: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: an alkyl group; or an alkenyl group; an aryl group; and aryl-lower alkyl; heteroaryl-lower alkyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R6: h; a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R7: a lower alkyl group; lower alkenyl; - (CH)2)qOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)qSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)qNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)qN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)rCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)qCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)rPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); (CH)2)rSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R9: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R10: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R11: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R12: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)rCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)rCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)rPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R13: a lower alkyl group; lower alkenyl; - (CH)2)qOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)qSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)qNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)rCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)qCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)rPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)rSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R14: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R15: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-；Or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); particularly preferred is NR20CO lower alkyl (R)20= H; or lower alkyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R16: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R17: a lower alkyl group; lower alkenyl; - (CH)2)qOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)qSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)qNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)qN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)rCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)qCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)rPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)rSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
Among the structural units a1 to a69, the following are preferred: r2A5, A8, A22, A25, R for H2A38, A42, A47, and A50 for H. Most preferred is a structural element of type A8':
wherein R is20Is H or lower alkyl; and R64Is an alkyl group; an alkenyl group; an aryl group; aryl-lower alkyl; or heteroaryl-lower alkyl; especially wherein R64Is n-hexyl (A8' -1); n-heptyl (A8' -2); 4- (phenyl) benzyl (A8' -3); diphenylmethyl (A8' -4); 3-amino-propyl (A8' -5); 5-amino-pentyl (A8' -6); methyl (A8' -7); ethyl (A8' -8); isopropyl (A8' -9); isobutyl (A8' -10); n-propyl (A8' -11); cyclohexyl (A8' -12); cyclohexylmethyl (A8' -13); n-butyl (A8' -14); phenyl (A8' -15); benzyl (A8' -16); (3-indolyl) methyl (A8' -17); 2- (3-indolyl) ethyl (A8' -18); (4-phenyl) phenyl (A8' -19); and n-nonyl (A8' -20).
The structural unit A70 belongs to the class of open-chain alpha-substituted alpha-amino acids, the structural units A71 and A72 to the corresponding beta-amino acid analogs and the structural units A73-A104 to the cyclic analogs of A70. These amino acid derivatives have been shown to confine small peptides in a well-defined reverse-turn or U-shaped conformation (C.M. Venkatachalam, Biopolymers, 1968, 6, 1425-one 1434; W.Kabsch, CSander, Biopolymers, 1983, 22, 2577). These Building blocks or templates are ideally suited for stabilizing the beta-hairpin conformation in the Peptide loop (D.Obrecht, M.Altorfer, J.A.Robinson, "Novel Peptide Mimetic Building blocks and Strategies for Efficient guidance of search), adv.Med chem.1999, Vol.4, 1-68; P.Balaram," Non-standard amino acids in Peptide design and protein engineering, "curr.Opin Struct.biol.1992, 2, 845, 851; M.Crisma, G.Valle, C.Toniolo, S.asaad, R.B.Rao, P.Balaram," alpha-disubstituted amino acid containing models, Al. J.249, V.J.1990; Peptide conformation models, Al. Biochem.35-J.35, O.J.35, O.J.249, J.Pat.Biochem.78, et al., 1-68).
It has been found that two enantiomers of the building Blocks-A70-CO-to A104-CO-in combination with the L-configured building block-B-CO-can effectively stabilize and induce beta-hairpin conformations (D.Obrecht, M.Altorfer, J.A.Robinson, "Novel Peptide mimetic building Blocks and Strategies for Efficient guidance in search), adv.MedChem.1999, Vol.4, 1-68; D.Obrecht, C.Spiegler, P.K.Müller，H.Heimgartner，F.Stierli，Helv.Chim.Acta 1992，75，1666-1696；D.Obrecht，U.Bohdal，J.Daly，C.Lehmann，P.K.Müller，Tetrahedron，1995，51，10883-10900；D.Obrecht，C.Lehmann，C.Ruffieux，P.K.Müller，Helv.Chim.Acta 1995，78，1567-1587；D.Obrecht，U.Bohdal，C.Broger，D.Bur，C.Lehmann，R.Ruffieux，P.C.Spiegler，Helv.Chim.Acta1995，78，563-580；D.Obrecht，H.Karajiannis，C.Lehmann，P.C.Spiegler，Helv.Chim.Acta1995，78，703-714）。
Thus, for the purposes of the present invention, template (A1) may also consist of the structural units-A70-CO-to A104-CO-in which the structural units A70 to A104 are in the (D) -or (L) -configuration, in combination with the structural unit-B-CO-in the (L) -configuration.
R in A70-A10420Preferred values of (b) are H or lower alkyl, with methyl being most preferred. R in the structural units A70 to A10418，R19And R21-R29Preferred values of (a) are as follows:
-R18: a lower alkyl group.
-R19: a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; r33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)pCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)pCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)pSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)oC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R21: h; a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: low gradeAn alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: low gradeAn alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R22: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R23: h; a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl)；-(CH2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); particularly preferred is NR20CO lower alkyl (R)20= H; or lower alkyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy);
-R24: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); particularly preferred is NR20CO lower alkyl (R)20= H; or lower alkyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R25: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R26: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lowA lower alkyl group); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-or, R25And R26Taken together may be- (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or a lower alkyl group.
-R27: h; a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R28: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl(ii) a Or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R29: a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); particularly preferred is NR20CO lower-alkyl (R)20= H; or lower alkyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
For templates (b) through (p), such as (b 1) and (c 1), preferred values for the various symbols are as follows:
-R8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R20: h; or a lower alkyl group.
-R30: h, methyl.
-R31: h; a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; r33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-；R57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)rC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy). Most preferred is-CH2CONR58R59(R58H, or lower alkyl; r59: lower alkyl or lower alkenyl);
-R32: h, methyl.
-R33: a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR34R63(wherein R is34: a lower alkyl group; or lower alkenyl; r63: h; or lower alkyl; or R34And R63Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR75R82(wherein R is75: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R75And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR78R82(wherein R is20: h; or lower alkyl; r78: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R78And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl).
-R34: h; or a lower alkyl group.
-R35: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl).
-R36: a lower alkyl group; lower alkenyl; or aryl-lower alkyl.
-R37: h; a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R38: h; a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenesA group; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R39: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl).
-R40: a lower alkyl group; lower alkenyl; or aryl-lower alkyl.
-R41: h; a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R42: h; a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R43: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57：H；Or lower alkyl); - (CH)2)oPO(OR60)2(wherein R is60: a lower alkyl group; or lower alkenyl); - (CH)2)oSO2R62(wherein R is62: a lower alkyl group; or lower alkenyl); or- (CH)2)qC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R44: a lower alkyl group; lower alkenyl; - (CH)2)pOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)pSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)pNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R78Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)pN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)pCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)pCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)oC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R45: h; a lower alkyl group; lower alkenyl; - (CH)2)oOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)oSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)oNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)sOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)oN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)sC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R46: h; a lower alkyl group; lower alkenyl; - (CH)2)sOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)sSR56(wherein R is56: a lower alkyl group; or lower alkenyl); - (CH)2)sNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)sOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)sNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)sN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)sC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R47: h; OR OR55(wherein R is55: a lower alkyl group; or lower alkenyl).
-R48: h; or a lower alkyl group.
-R49: h; a lower alkyl group; - (CH)2)oCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)oCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)sC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R50: h; a methyl group.
-R51: h; a lower alkyl group; lower alkenyl;-(CH2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)pCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)pCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; andR59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)rC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R52: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; or lower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)pCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)pCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)rC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R53: h; a lower alkyl group; lower alkenyl; - (CH)2)mOR55(wherein R is55: a lower alkyl group; or lower alkenyl); - (CH)2)mNR33R34(wherein R is33: a lower alkyl group; or lower alkenyl; r34: h; or lower alkyl; or R33And R34Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mOCONR33R75(wherein R is33: h; or lower alkyl; orLower alkenyl; r75: a lower alkyl group; or R33And R75Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mNR20CONR33R82(wherein R is20: h; or lower alkyl; r33: h; or lower alkyl; or lower alkenyl; r82: h; or lower alkyl; or R33And R82Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); - (CH)2)mN(R20)COR64(wherein: R20: h; or lower alkyl; r64: a lower alkyl group; or lower alkenyl); - (CH)2)pCOOR57(wherein R is57: a lower alkyl group; or lower alkenyl); - (CH)2)pCONR58R59(wherein R is58: a lower alkyl group; or lower alkenyl; and R59: h; or lower alkyl; or R58And R59Together form: - (CH)2)2-6-；-(CH2)2O(CH2)2-；-(CH2)2S(CH2)2-; or- (CH)2)2NR57(CH2)2-; wherein R is57: h; or lower alkyl); or- (CH)2)rC6H4R8(wherein R is8：H；F；Cl；CF3(ii) a A lower alkyl group; lower alkenyl; or lower alkoxy).
-R54: a lower alkyl group; lower alkenyl; or aryl-lower alkyl.
Among the structural units a70 to a104, the following are preferred: r22A74, A75, A76, R for H22A77, a78 and a79 for H.
The structural units-B-CO-in templates (a1) and (a2) represent L-amino acid residues. Preferred values for B are: -NR20CH(R71) -and R2The corresponding isomers of the group A5 of H, A8, A22, A25, R2A38, A42, A47, and A50 for H. Most preferred is
Furthermore, the most preferred values of B also include (L) configuration of a group of type A8 ":
wherein R is20Is H or lower alkyl and R64Is an alkyl group; an alkenyl group; - [ (CH)2)u-X]t-CH3(wherein X is-O-; -NR-)20-; or-S-; u =1-3, and t = 1-6), aryl; aryl-lower alkyl; or heteroaryl-lower alkyl; especially those radicals in which R64Is n-hexyl (A8' -21); n-heptyl (A8 "-22); 4- (phenyl) benzyl (A8 "-23); diphenylmethyl (A8 "-24); 3-amino-propionic acidA radical (A8' -25); 5-amino-pentyl (A8 '-26; methyl (A8' -27); ethyl (A8 '-28); isopropyl (A8' -29); isobutyl (A8 '-30); n-propyl (A8' -31); cyclohexyl (A8 '-32); cyclohexylmethyl (A8' -33); n-butyl (A8 '-34); phenyl (A8' -35); benzyl (A8 '-36); 3-indolyl) methyl (A8' -37); 2- (3-indolyl) ethyl (A8 '-38); 4-phenyl) phenyl (A8' -39); and n-nonyl (A8 '-40); CH 8' -27); and3-OCH2CH2-OCH2- (A8 "-41) and CH3-(OCH2CH2)2-OCH2-(A8”-42)。
The peptide chain Z of the beta-hairpin mimetics described herein is generally defined in terms of amino acid residues belonging to one of the following groups
Group C-NR20CH(R72) CO-; "hydrophobic: small to medium size "
Group D-NR20CH(R73) CO-; "hydrophobic: large aromatic or heteroaromatic compounds "
Group E-NR20CH(R74) CO-; "polar-cationic" and "urea-derived"
Group F-NR20CH(R84) CO-; "polar-uncharged or anionic"
Group H-NR20-CH(CO-)-(CH2)4-7-CH(CO-)-NR20-；
-NR20-CH(CO-)-(CH2)pSS(CH2)p-CH(CO-)-NR20-；
-NR20-CH(CO-)-(-(CH2)pNR20CO(CH2)p-CH(CO-)-NR20-; and
-NR20-CH(CO-)-(-(CH2)pNR20CONR20(CH2)p-CH(CO-)-NR20-；
"interchain bond"
In addition, the amino acid residues in the chain Z may also be of the formula-A-CO-or of the formula-B-CO-, wherein A and B are as defined above. Finally, Gly may also be an amino acid residue in chain Z, and Pro may also be an amino acid residue in chain Z, except for positions where an interchain linkage (H) may be present.
Group C includes groups according to the para-substituent R72With small to medium-sized hydrophobic side chain groups. Hydrophobic residues refer to amino acid side chains that are uncharged at physiological pH and are rejected by aqueous solutions. In addition, these side chains generally do not contain hydrogen bond donor groups such as, but not limited to, primary and secondary amides, primary and secondary amines and their corresponding protonated salts, thiols, alcohols, phosphonates, phosphates, ureas or thioureas. However, they may contain hydrogen bond acceptor groups, such as ethers, thioethers, esters, tertiary amides, alkyl-or arylphosphonates and phosphates or tertiary amines. Gene-encoded small to medium-sized amino acids include alanine, isoleucine, leucine, methionine and valine.
Group D includes groups according to the para-substituent R73Have aromatic and heteroaromatic side chain groups. An aromatic amino acid residue refers to a hydrophobic amino acid having a side chain comprising at least one ring with a conjugated pi-electron system (aromatic group). They may additionally contain hydrogen bond donor groups such as, but not limited to, primary and secondary amides, primary and secondary amines and their corresponding protonated salts, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as, but not limited to, ethers, thioethers, esters, tertiary amides, alkyl-or arylphosphonates-and phosphates or tertiary amines. Aromatic amino acids encoded by the gene include phenylalanine and tyrosine.
Heteroaromatic amino acid residues are referred to in terms of the para-substituent R77Has a side chain comprising at least one ring having a conjugated pi wherein at least one heteroatom such as (but not limited to) O, S and N is introduced-a system. In addition, such residues may contain hydrogen bond donor groups such as, but not limited to, primary and secondary amides, primary and secondary amines and their corresponding protonated salts, thiols, alcohols, phosphonates, phosphates, ureas or thioureas, and hydrogen bond acceptor groups such as, but not limited to, ethers, thioethers, esters, tertiary amides, alkyl-or arylphosphonates and phosphates or tertiary amines. The gene encodes heteroaromatic amino acids including tryptophan and histidine.
Group E includes groups according to the para-substituent R74Contains amino acids with side chains of polar-cationic, amido-and urea-derived residues. Polar-cationic refers to a basic side chain that is protonated at physiological pH. The polar-cationic amino acids encoded by the gene include arginine, lysine and histidine. Citrulline is an example of a urea-derived amino acid residue.
Group F includes groups according to the para-substituent R84Comprises amino acids having side chains with polar-uncharged or anionic residues. Polar-uncharged or anionic residues refer to hydrophilic side chains that are uncharged at physiological pH and are anionic in nature (carboxylic acids included), but are not repelled by aqueous solutions. These side chains typically contain hydrogen bond donor groups such as, but not limited to, primary and secondary amides, carboxylic acids and esters, primary and secondary amines, thiols, alcohols, phosphonates, phosphates, ureas, or thioureas. These groups can form a hydrogen bonding network with water molecules. In addition, they may also contain hydrogen bond acceptor groups such as, but not limited to, ethers, thioethers, esters, tertiary amides, carboxylic acids and carboxylates, alkyl-or arylphosphonate-and phosphate esters or tertiary amines. The polar-uncharged amino acids encoded by the gene include asparagine, cysteine, glutamine, serine and threonine, but may also be aspartic acid and glutamic acid.
Group H comprises side chains of preferably (L) -amino acids at opposite positions of the β -strand region where interchain bonds can be formed. The most common bond is a disulfide bridge formed by cysteine and homocysteine located in opposite positions of the beta-strand. Various methods are known for forming disulfide bonds, including those described below: tam et al, Synthesis, 1979, 955-957; stewart et al, Solid Phase Peptide Synthesis, 2 nd edition, Pierce Chemical Company, III., 1984; ahmed et al, J.biol.chem.1975, 250, 8477-8482; and Pennington et al, Peptides, p164-166, Giralt and Andreu editions, ESCOMEIDEN, the Netherlands, 1990. Most advantageously, for the scope of the present invention, an acetamidomethyl (Acm) -protecting group (for cysteine) can be used to make the disulfide bond. One defined interchain linkage is formed by the amide bond formation of ornithine and lysine, respectively, to glutamic acid and aspartic acid residues at opposite β -strand positions. For aspartic acid and glutamic acid, preferred protecting groups for the pendant amino-groups of ornithine and lysine are allyloxycarbonyl (Alloc) and allyl ester. Finally, interchain linkages may also be established by linking the amino groups of lysine and ornithine located at opposite β -strand positions with a reagent such as N, N-carbonyl imidazole to form a cyclic urea.
As previously mentioned, the positions of the interchain bonds are the P4 and P9 positions together; and/or P2 and P11. These interchain linkages are known to stabilize the β -hairpin conformation and thus constitute an important structural component for the design of β -hairpin mimetics.
The most preferred amino acid residues in the chain Z are those derived from natural alpha-amino acids. The amino acids themselves or residues thereof suitable for the purposes of the present invention are listed below, where abbreviations correspond to the commonly used convention:
other α -amino acids or residues thereof suitable for the purposes of the present invention include:
particularly preferred residues of group C are:
particularly preferred residues of group D are:
particularly preferred residues of group E are:
particularly preferred residues of group F are
Typically, the peptide chain Z within the β -hairpin mimetics of the invention comprises 12 amino acid residues. The positions P1-P12 of each amino acid residue in the chain Z are clearly defined as follows: p1 represents the first amino acid in chain Z coupled at its N-terminus to the C-terminus of template (B) - (P) or of group-B-CO-in template (a1), or of group-A-CO-in template (a 2); p12 represents the last amino acid in chain Z coupled at its C-terminus to the N-terminus of template (B) - (P) or of group-A-CO-in template (a1) or of group-B-CO-in template (a 2). Positions P1 to P12 each preferably contain an amino acid residue belonging to one of the above groups C, D, E, F, H, or an amino acid residue of the formula-A-CO-or of the formula-B-CO-, or Gly, or Pro, as follows:
the α -amino acid residues in positions 1 to 12 of the chain Z are preferably:
-P1: type C or type D or type E or type F;
-P2: type D;
-P3: type C, or residue Gly or Pro;
-P4: type C, or type E, or type F, or the residue is Gly or Pro;
-P5: type E, or residue Gly or Pro;
-P6: type E, type C or type F, or of the general formula-A-CO-, or the residue is Gly or Pro;
-P7: type C, type E, or type F, or of the general formula-B-CO-;
-P8: type D, or type F;
-P9: type E, or type F or type C;
-P10: type E;
-P11: type F or type C, or residue is Gly or Pro; and
-P12: type C or type D or type E or type F; or
-P4 and P9 and/or P2 and P11 together may form a group of type H; and at P6, P10 and P11, also the D-isomer;
or, alternatively, in the less preferred embodiments described above:
-P1: type C or type D or type E or type F;
-P2: type F or type C, or residue is Gly or Pro;
-P3: type E;
-P4: type E, or type F, or type C;
-P5: type D, or type F;
-P6: type C, type E or type F, or of the general formula-B-CO-;
-P7: type C, type F, or formula-A-CO-, or the residue is Gly or Pro;
-P8: type E, or residue Gly or Pro;
-P9: type C, or type E or type F, or the residue is Gly or Pro;
-P10: type C, or residue Gly or Pro;
-P11: type D; and
-P12: type C or type D or type E or type F; or
-P4 and P9 and/or P2 and P11 together may form a group of type H; and at P2, P3 and P7, also the D-isomer;
if n is 12, the most preferred alpha-amino acid residues in positions 1 to 12 are:
-P1：Ala、Cit、Thr、Thr、Asp、Glu；
-P2：Trp、Tyr；
-P3：Ile、Val、Nle、Chg、Cha；
-P4：Dab、Lys、Gln；
-P5：Lys、Dab、Orn；
-P6：Dab、DDab；Lys；
-P7：His、Lys、Gln、Dab；
-P8：Tyr、Trp、Ser；
-P9：Dab、Lys；
-P10：Dab、Lys；
-P11：Ala、Abu、Thr、Gly、Pro、Hse、Ile、Nva、DAla、DVal、Aib、Nle、Chg、Cha、Gln、Asp、Glu、Cpa、t-BuG、Leu、Val、Asn；
-P12：Dab、Lys、Gln、Ser；
it may also be the D isomer at P6, P10 and P11.
Particularly preferred β -hairpin peptidomimetics of the invention include those described in examples 1,2, 6, 16, 19, 22, 24, 25, 28, 29, 32, 35, 40, 41, 49, 50.
The methods of the invention can advantageously be carried out according to parallel array synthesis (parallel array synthesis) to generate a library of template-fixed β -hairpin peptidomimetics of formula I above. These parallel syntheses can yield multiple (usually 24-192, typically 96) arrays of compounds of formula I in high yields and defined purity, with minimal formation of dimer and polymer by-products. Thus, proper selection of template and cyclization reaction sites plays an important role for the functionalized solid support (i.e., solid support plus linker molecule).
Polystyrene crosslinked with preferably 1-5% divinylbenzene; polystyrene coated with polyethylene glycol spacers (Tentagel)R) (ii) a And polyacrylamide resins (see also Obrecht, D.; Villalgord, J. -M, "Solid-Supported Combinatorial and parallel Synthesis of Small-Molecular-Weight Compounds libraries)," Tetrahedron Organic Chemistry Series, Vol.17, Pergamon, Elsevierscience, 1998).
The solid support is functionalized with a linker, i.e., a bifunctional spacer molecule containing an anchoring group at one end for attachment to the solid support and a selectively cleavable functional group at the other end for subsequent chemical conversion and cleavage steps. For the purposes of the present invention, two types of linkers are used:
type 1 linkers are designed to release the amide group under acidic conditions (Rink H, Tetrahedron Lett.1987, 28, 3783-. This type of linker forms an amide of the amino acid carboxyl group; examples of resins functionalized by such linker structures include 4- [ ((((2, 4-dimethoxyphenyl) Fmoc-aminomethyl) phenoxyacetamide) aminomethyl ] PS resin, 4- [ (((2, 4-dimethoxyphenyl) Fmoc-aminomethyl) phenoxyacetamide) aminomethyl ] -4-methylbenzhydrylamine PS-resin (Rink amide MBHA PS resin) and 4- [ ((((2, 4-dimethoxyphenyl) Fmoc-aminomethyl) phenoxyacetamide) aminomethyl ] benzhydrylamine PS resin (Rink amide bhps resin). Preferably, the support is derived from polystyrene cross-linked with divinylbenzene (most preferably 1-5%) and functionalized by a 4- [ (((2, 4-dimethoxyphenyl) Fmoc-aminomethyl) phenoxyacetamide) linker.
The type 2 linker was designed to eventually release the carboxyl group under acidic conditions. This type of linker forms acid labile esters with the carboxyl group of amino acids, typically acid labile benzyl, benzhydryl and trityl esters; examples of such linker structures include 2-methoxy-4-hydroxymethylphenoxy (Sasrin)RLinker), 4- (2, 4-dimethoxyphenyl-hydroxymethyl) -phenoxy (Rink linker), 4- (4-hydroxymethyl-3-methoxyphenoxy) butanoic acid (HMPB linker), trityl, and 2-chlorotrityl. Preferably, the support is derived from polystyrene cross-linked with divinylbenzene (most preferably 1-5%) and functionalized by 2-chlorotrityl linkers.
When performed as a parallel array synthesis, the process of the invention may advantageously be performed as described below, but it will be immediately apparent to the skilled person how these procedures may be adjusted as necessary when the synthesis of a single compound of formula I above is required.
The number of reaction vessels (generally 24-192, typically 96) is equal to the total number of compounds to be synthesized by the parallel method, to which 25-1000mg, preferably 100mg of a suitable functionalized solid support, preferably 1-3% cross-linked polystyrene or Tentagel resin, are loaded.
The solvent to be used must be capable of swelling the resin and includes, but is not limited to, Dichloromethane (DCM), Dimethylformamide (DMF), N-methylpyrrolidone (NMP), dioxane, toluene, Tetrahydrofuran (THF), ethanol (EtOH), Trifluoroethanol (TFE), isopropyl alcohol, and the like. Solvent mixtures comprising a polar solvent as at least one component (e.g., 20% TFE/DCM, 35% THF/NMP) are beneficial for ensuring high reactivity and solvation of resin-bonded peptide chains (Fields, G.B., Fields, C.G., J.Am.chem.Soc.1991, 113, 4202-4207).
Significant progress has been made in the synthesis of protected peptide fragments with the development of various linking agents that release the C-terminal carboxylic acid group under mild acidic conditions that do not affect the acid labile group protecting the functional group in the side chain. 2-methoxy-4-hydroxybenzyl alcohol-derived linkers (Sasrin)RLinkers, Mergler et al, Tetrahedron Lett., 1988, 294005-4008) can be cleaved with dilute trifluoroacetic acid (0.5-1% TFA in DCM) and stabilized against Fmoc deprotection conditions during peptide synthesis, where the Boc/t Bu-group attachment of a protecting group is compatible with the protection scheme. Suitable for the process of the inventionIts linkers include the superacid labile 4- (2, 4-dimethoxyphenyl-hydroxymethyl) -phenoxy linker (Rink linker, Rink, h. tetrahedron lett, 1987, 28, 3787-; 4- (4-hydroxymethyl-3-methoxyphenoxy) butanoic acid-derived linker (HMPB-linker,&riniker, Peptides, 1991, 1990131), can also be cleaved with 1% TFA/DCM to give peptide fragments containing all acid labile side chain protecting groups; and, in particular, the 2-chlorotrityl chloride linker (Barlos et al, Tetrahedron Lett., 1989, 30, 3943-.
Suitable protecting groups for amino acids and correspondingly for their residues are, for example,
for amino groups (if present, for example in the side chain of lysine)
For carboxyl groups (if present, for example in the side chains of aspartic acid and glutamic acid), conversion to esters by reaction with an alcohol component
For guanidino groups (if present, for example in the side chain of arginine)
For hydroxyl groups (if present, for example in the side chains of threonine and serine)
tBu tert-butyl
Bn benzyl group
Trt trityl radical
And for the thiol group (if present, for example in the side chain of cysteine)
The 9-fluorenylmethoxycarbonyl- (Fmoc) -protected amino acid derivatives are preferably used as building blocks for the construction of template-immobilized beta-hairpin loop mimetics of the general formula I. For deprotection, i.e. cleavage of the Fmoc group, 20% piperidine in DMF or 2% DBU/2% piperidine in DMF may be used.
The content of reactants, i.e. amino acid derivatives, is typically 1 to 20 equivalents based on the milliequivalents per gram (meq/g) loading of the functionalized solid support initially weighed into the reaction tube (typically 0.1-2.85meq/g for polystyrene resins). Additional equivalents of reactants are used, if necessary, to complete the reaction in a reasonable time. The reaction tubes, as well as the clamp set and manifold, are reinserted into the reservoir set and the device is secured together. A gas flow is initiated through the manifold to provide a controlled environment, such as nitrogen, argon, air, and the like. The gas flow may also be heated or cooled before flowing through the manifold. Heating or cooling of the reaction well is achieved by heating the reaction zone or external cooling with isopropanol/dry ice and the like to achieve the desired synthesis reaction. Agitation is achieved by shaking or magnetic stirring (within the reaction tube). Preferred workstations (not, but not limited to) are Labsource's Combi-chem station and MultiSynTech's-Syro synthesizers.
Amide bond formation requires activation of the alpha-carboxyl group for the acylation step. By means of customary carbodiimides, such as dicyclohexylcarbodiimide (DCC, Sheehan)&When Hess, J.Am.chem.Soc.1955, 77, 1067-1068) or diisopropylcarbodiimide (DIC, Saratakis et al, biochem.Biophys.Res.Commun.1976, 73, 336-342) were used for the reaction, dicyclohexylurea and diisopropylurea were obtained which were each insoluble and soluble in the usual solvents. In a variation of the carbodiimide method, 1-hydroxybenzotriazole (HOBt,&geiger, chem. Ber1970, 103, 788-. HOBt prevents dehydration, inhibits racemization of activated amino acids and acts as a catalyst to promote slow coupling reactions. Certain phosphonium reagents have been used as direct coupling reagents, such as benzotriazol-1-yl-oxy-tris- (dimethylamino) -phosphonium hexafluorophosphate (BOP, Castro et al, Tetrahedron Lett., 1975, 14, 1219-1222; Synthesis, 1976, 751-752), or benzotriazol-1-yl-oxy-tris-pyrrolidinyl-phosphonium hexafluorophosphate (Py-BOP, Coste et al, Tetrahedron Lett., 1990, 31, 205-208), or 2- (1H-benzotriazol-1-yl-) 1, 1,3, 3-tetramethyluronium tetrafluoroborate (TBTU), or hexafluorophosphate (HBTU, Knorr et al, Tetrahedron Lett., 1989, 30, 1927-1930); or 1-benzotriazole-1- [ bis (dimethylamino) methylene]-5-chloro-hexafluorophosphate-1, 3-oxide (HCTU); these phosphonium reagents are also suitable for forming HOBt esters in situ with protected amino acid derivatives. More recently, diphenoxyphosphoryl azide (DPPA) or O- (7-aza-benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium tetrafluoroborate (TATU) or O- (7-aza-benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU)/7-aza-1-hydroxybenzotriazole (HOAt, Carpino et al, tetrahedron Lett., 1994, 35, 2279-.
Due to the fact that near quantitative coupling reactions are necessary, it is desirable to have experimental evidence of reaction completion. Ninhydrin tests (Kaiser et al, anal. biochemistry, 1970, 34, 595) can be easily and quickly performed after each coupling step, where a positive colorimetric response to an aliquot of the resin-bound peptide qualitatively indicates the presence of a primary amine. Fmoc chemistry enables the detection of Fmoc chromophores spectrophotometrically when released with bases (Meienhofer et al, int.J. peptide Protein Res.1979, 13, 35-42).
The resin bound intermediate within each reaction tube is washed by one of two methods, excess retained reagent, solvent and by-products by repeated exposure to pure solvent:
1) filling the reaction well (preferably 5 ml) with solvent, immersing and stirring the reaction tube with the set of clamps and manifold for 5 to 300 minutes, preferably 15 minutes, and then draining by gravity, followed by removing the solvent by applying air pressure through the manifold inlet (while closing the outlet);
2) the manifold is removed from the jig set and an aliquot (preferably 5 ml) of the solvent is dispensed through the top of the reaction tube and discharged by gravity through a filter into a receiving container such as a test tube or vial.
The above washing steps are repeated up to about 50 times (preferably about 10 times) with the removal efficiency of the reagents, solvents and by-products monitored by methods such as TLC, GC, or inspection of the washings.
The above procedure of reacting the resin bound compound with the reagent in the reaction well and then removing excess reagent, by-products and solvent is repeated for each successive conversion process until the final resin bound fully protected linear peptide is obtained.
Before the fully protected linear peptide is detached from the solid support, one or more protected functional groups present in the molecule may be selectively deprotected and the reactive groups thus liberated suitably substituted, if desired. For this purpose, the functional groups must initially be protected by protecting groups which can be removed selectively without affecting the processDepending on the remaining protecting groups present. Alloc (allyloxycarbonyl) is an example of such an amino protecting group which can be selectively removed, for example, by CH2Cl2Pdo and phenylsilane without affecting the remaining protecting groups present in the molecule, such as Fmoc. The reactive groups thus released may subsequently be treated with reagents suitable for introducing the desired substituents. Thus, for example, an amino group is acylated by an acylating agent corresponding to the acyl substituent to be introduced. For the formation of PEGylated amino acids such as I PegK or SPegK, it is preferred to add 5 equivalents of HATU (N- [ (dimethylamino) -1H-1,2, 3-triazolone [4,5-b ]]Pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide) and 10 equivalents of DIPEA (diisopropylethylamine) and 5 equivalents of 2- [2- (2-methoxyethoxy) ethoxy ] ethoxy]Acetic acid (IPeg), and respectively 2- (2-methoxyethoxy) acetic acid (sPeg), were applied to the amino group released from the side chain of the appropriate amino acid for 3 h. After filtering and washing the resin afterwards, the procedure was repeated for another 3h with fresh reagent solution.
Before detaching the fully protected linear peptide from the solid support, interchain bonds may also be formed between the side chains of appropriate amino acid residues in the relative positions of the β -strand fragments, if desired.
The interchain bonds and their formation are discussed above, as well as explanations with respect to group H of the type, which may be, for example, disulfide bridges formed by cysteine and homocysteine in the relative positions of the β -strands; or by glutamic acid and aspartic acid, each of which links an ornithine and a lysine residue, or by amide bond formation of a lactam bridge linking a glutamic acid residue of a2, 4-diaminobutyric acid residue located at the opposite beta-chain position. Such interchain bond formation can be achieved by methods well known in the art.
For the formation of disulfide bridges, preferably 10 equivalents of iodine solution are applied to DMF or CH2Cl2In the MeOH mixture for 1.5h, the iodine solution was filtered and repeated for another 3h with fresh iodine solution, or in the DMSO and acetic acid mixture with 5% NaHCO3Buffered to pH5-6 for 4h, or stirred in water after having been adjusted to pH8 with ammonium hydroxide solution for 24h, or ammonium acetate buffer adjusted to pH8 in the presence of air, or in a solution of NMP and tri-n-butyl phosphine (preferably 50 equivalents).
The separation of the fully protected linear peptide from the solid support is achieved by dipping the reaction tube, as well as the set of clamps and manifold into a reaction well containing a solution of the cleaving reagent (preferably 3 to 5 ml). Gas flow, temperature control, agitation, and reaction monitoring are as described above and performed as needed to effect the isolation reaction. The reaction tubes, as well as the clamp set and manifold, are disconnected from the reservoir set and raised above the solution level but below the upper lip of the reaction wells, and then air pressure is applied through the manifold inlet (while closing the outlet) to effectively expel the final product solution into the reservoir wells. The resin remaining in the reaction tube is then washed 2 to 5 times as above with 3 to 5ml of a suitable solvent to extract (wash out) as much of the isolated product as possible. The product solutions thus obtained were combined, taking care to avoid cross-mixing. Each solution/extract is then treated as necessary to isolate the final compound. Typical treatments include, but are not limited to, evaporation, concentration, liquid/liquid extraction, acidification, basification, neutralization or other reactions in solution.
The solution containing the fully protected linear peptide derivative which has been separated from the solid support and neutralized with a base is evaporated. The cyclization reaction is then carried out in solution using solvents such as DCM, DMF, dioxane, THF and the like. Various coupling reagents mentioned earlier may be used for the cyclization reaction. The duration of the cyclization reaction is about 6 to 48 hours, preferably about 16 hours. The reaction progress was followed by RP-HPLC (reverse phase high Performance liquid chromatography). The solvent is then removed by evaporation, the fully protected cyclic peptide derivative is dissolved in a water-immiscible solvent, such as DCM, and the solution is extracted with water or a mixture of water-miscible solvents to remove any excess coupling reagent.
Finally, with 95% TFA, 2.5% H2O, 2.5% TIS or another scavenger combination to treat fully protected peptide derivativesCleavage of the protecting group. The cleavage reaction time is generally 30 minutes to 12 hours, preferably about 2.5 hours. The volatiles were evaporated to dryness and the crude peptide was dissolved in 20% AcOH in water and extracted with isopropyl ether or other suitable solvent. The aqueous layer was collected and evaporated to dryness to obtain the fully protected cyclic peptide derivative of formula I as the final product. Depending on its purity, the peptide derivative can be used directly in the biological analysis or must be further purified, for example by preparative HPLC.
Alternatively, separation of the fully protected peptide from the solid support, cyclization and complete deprotection can be achieved manually in glass containers.
The fully deprotected product of the general formula I thus obtained can then, as described before, if desired, be subsequently converted into a pharmaceutically acceptable salt or the pharmaceutically acceptable or unacceptable salt thus obtained into the corresponding free compound of the general formula I or into a different pharmaceutically acceptable salt. Any of these operations may be performed by methods well known in the art.
The template starting materials of the general formula II used in the process of the invention, the prepresses used therefor and the preparation of these starting materials and prepresses are described in International application PCT/EP02/01711, published as WO02/070547A 1.
The β -hairpin peptidomimetics of the invention can be used in a variety of applications to inhibit the growth of or kill microorganisms. In particular, they can be used to selectively inhibit the growth of or kill microorganisms such as Pseudomonas aeruginosa.
For example, they may be used as bactericides or preservatives in substances such as foodstuffs, cosmetics, pharmaceuticals and other nutrient-containing substances. The beta-hairpin peptidomimetics of the invention may also be used for the treatment or prevention of diseases associated with microbial infections of plants and animals.
For use as an antiseptic or preservative, the β -hairpin peptidomimetic can be added to the desired substance alone, as a mixture of several β -hairpin peptidomimetics, or in combination with other antimicrobial agents. The β -hairpin peptidomimetic can be administered as such or as a suitable formulation containing carriers, diluents, or excipients well known in the art.
When used to treat or prevent infections or diseases associated with such infections, particularly infections associated with respiratory diseases such as cystic fibrosis, emphysema and asthma; infections associated with skin or soft tissue diseases such as surgical wounds, wounds and burns; infections associated with gastrointestinal diseases such as epidemic diarrhea, necrotizing enterocolitis and cecececronitis; infections associated with ocular diseases such as keratitis and endophthalmitis; infections associated with otic disorders such as otitis; infections associated with CNS diseases such as brain abscesses and meningitis; infections associated with bone diseases such as osteochondritis and osteomyelitis; infections associated with cardiovascular diseases such as endocarditis and pericarditis; infections associated with gastrourinal diseases such as epididymitis, prostatitis, and urethritis can be administered alone, as a mixture of several β -hairpin peptidomimetics, in combination with other antimicrobial or antibiotic agents, or anti-cancer agents, or anti-viral (e.g., anti-HIV) agents, or in combination with other pharmaceutically active agents. The β -hairpin peptidomimetics can be administered as such or as a pharmaceutical composition.
Pharmaceutical compositions comprising the β -hairpin peptidomimetics of the invention may be prepared by conventional mixing, dissolving, granulating, coated tablet preparing, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. The pharmaceutical compositions may be formulated in conventional manner using one or more physiologically acceptable carriers, diluents, excipients or auxiliaries which facilitate processing of the active β -hairpin peptidomimetics into pharmaceutically acceptable preparations. The appropriate formulation depends on the method of administration chosen.
For topical administration, the beta-hairpin peptidomimetics of the present invention can be formulated into solutions, gels, ointments, creams, suspensions, and the like, as is well known in the art.
Systemic formulations include those designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral, or pulmonary administration.
For injection, the β -hairpin peptidomimetics of the invention are formulated in aqueous solution, preferably in a physiologically compatible buffer, such as Hink's solution, Ringer's solution, or physiological saline buffer. The solution may contain formulating agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the β -hairpin peptidomimetics of the invention may be in powder form for admixture with a suitable carrier, e.g., sterile pyrogen-free water, prior to use.
For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation, as is known in the art.
For oral administration, the mixture can be readily formulated by combining the active β -hairpin peptidomimetics of the invention with pharmaceutically acceptable carriers well known in the art. These carriers enable the beta-hairpin peptidomimetics of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. For oral formulations, for example, powders, capsules and tablets, suitable excipients include fillers, such as sugars, for example lactose, sucrose, mannitol and sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP); granulating agent; and a binding agent. If desired, disintegrating agents, such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate, may be added. The solid dosage forms may be sugar coated or enteric coated using standard techniques, if desired.
For oral liquid preparations, e.g., suspensions, elixirs and solutions, suitable carriers, excipients or diluents include water, glycols, oils, alcohols and the like. In addition, flavoring agents, preservatives, coloring agents, and the like may be added.
For oral administration, the compositions may be in the form of tablets, lozenges, and the like, formulated conventionally.
For administration by inhalation, the β -hairpin peptidomimetics of the invention are conveniently delivered as an aerosol spray from pressurized packs or a nebulizer, with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, carbon dioxide, or another suitable gas. In the case of a pressurised aerosol, the dosage unit may be measured by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the β -hairpin peptidomimetic of the invention and a suitable powder base such as lactose or starch.
The compounds may also be formulated in rectal or vaginal compositions, such as suppositories, with suitable suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the β -hairpin peptidomimetics of the invention can also be formulated as stock formulations. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. To prepare such depot formulations, the β -hairpin peptidomimetics of the invention may be formulated with suitable polymeric or hydrophobic substances (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as hardly soluble salts.
In addition, other drug delivery systems may be used, such as liposomes and emulsions, as are well known in the art. Specific organic solvents, such as dimethyl sulfoxide, may also be used. In addition, the β -hairpin peptidomimetics of the present invention can be delivered using a sustained release system such as a semipermeable matrix of a solid polymer containing the therapeutic agent. Various sustained release materials are established and are well known to those skilled in the art. Sustained release capsules can release the compound for weeks to over 100 days depending on its chemical nature. Other techniques for protein stabilization may be used depending on the chemical nature and biological stability of the therapeutic agent.
Because the β -hairpin peptidomimetics of the invention may contain charged residues, they may be included in any of the formulations described above as such or as a pharmaceutically acceptable salt. Pharmaceutically acceptable salts tend to be more soluble in aqueous and other protic solvents than the corresponding free base form.
The β -hairpin peptidomimetics of the invention, or compositions thereof, are generally used in an effective amount to achieve the intended purpose. It will be appreciated that the amount depends on the particular application.
For use as a bactericide or preservative, for example, an antimicrobially effective amount of a β -hairpin peptidomimetic of the invention or a composition thereof is applied to or added to the substance to be sterilized or preserved. An antimicrobially effective amount refers to the amount of the β -hairpin peptidomimetics or compositions thereof of the invention that inhibits the growth of the target microbiota, or the lethal amount thereof. Although an antimicrobially effective amount depends on the particular application, for use as a bactericide or preservative, the β -hairpin peptidomimetics of the invention or compositions thereof are generally added or applied in relatively low amounts to the substance to be sterilized or preserved. Generally, the β -hairpin peptidomimetics of the invention contain less than about 5% by weight, preferably less than 1% by weight and more preferably less than 0.1% by weight of the germicidal solution or substance to be preserved. One of ordinary skill in the art can use, for example, the in vitro assays provided in the examples to determine an antimicrobially effective amount of a particular β -hairpin peptidomimetic of the invention for a particular application without undue experimentation.
For use in treating or preventing microbial infections or diseases associated with such infections, the β -hairpin peptidomimetics of the invention or compositions thereof are administered or administered in a therapeutically effective amount. A therapeutically effective amount is an amount effective to ameliorate symptoms of, or ameliorate, treat or prevent a microbial infection or a disease associated therewith. Determination of a therapeutically effective amount is well within the ability of those skilled in the art, especially in view of the detailed description provided herein.
As with the antimicrobial and preservative agents, for topical administration for the treatment or prevention of bacterial infections, the therapeutically effective amount can be determined using, for example, the in vitro assays provided in the examples. Treatment may be performed when the infection is visible, or even when it is not visible. One of ordinary skill in the art is able to determine a therapeutically effective amount for treating a topical infection without undue experimentation.
For systemic administration, the therapeutically effective amount is initially estimated from an in vitro assay. For example, a dose can be formulated in animal models to achieve a circulating β -hairpin peptidomimetic concentration range that includes the IC as determined in cell culture50(i.e., 50% of the concentration of test compound lethal to the cell culture), as determined by the MIC in the cell culture (i.e., 100% of the concentration of test compound lethal to the cell culture). This information can be used to more accurately determine the dosage available to a person.
The starting dose can also be determined from in vivo data, such as animal models, using techniques well known in the art. One of ordinary skill in the art can readily optimize administration to humans based on animal data.
The dosage used as an antimicrobial agent can be adjusted individually to provide plasma levels of the β -hairpin peptidomimetics of the invention sufficient to maintain therapeutic effect. Therapeutically effective serum levels can be achieved by multiple daily administrations.
In the case of local administration or selective absorption, the effective local concentration of the β -hairpin peptidomimetics of the invention may not be correlated with the plasma concentration. One skilled in the art would be able to optimize therapeutically effective topical dosages without undue experimentation.
The amount of β -hairpin peptidomimetic to be administered will of course depend on the patient to be treated, the weight of the patient, the severity of the disease, the mode of administration and the judgment of the prescribing physician.
Antimicrobial treatments may be repeated intermittently when infections are detectable or even when they are not. Treatment may be provided alone or in combination with other drugs such as, for example, antibiotics or other antimicrobial agents.
Generally, a therapeutically effective amount of a β -hairpin peptidomimetic as described herein will provide therapeutic benefit without causing significant toxicity.
Haemolysis of erythrocytes is commonly used to assess the toxicity of related compounds such as the proteolytic enzyme lysin or tachplesin. Values are given as% lysis of erythrocytes observed at a concentration of 100. mu.g/ml. Typical values for cationic peptides such as the lysins and tachplesins are 30-40% for a wide range of pathogens, with an average MIC-value of 1-5. mu.g/ml. In general, the β -hairpin peptidomimetics of the invention have haemolytic values in the range of 0.5-10%, typically in the range of 1-5%, at levels of activity corresponding to those described above for lysins and tachplesins. Thus, preferred compounds have a low MIC-value and low% hemolysis of erythrocytes observed at a concentration of 100. mu.g/ml.
Toxicity of the beta-hairpin peptidomimetics of the invention can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by measuring LD50(lethal dose of 50% of population) or LD100(lethal dose of 100% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index. Compounds exhibiting a high therapeutic index are preferred. The data obtained from these cell culture assays and animal studies can be used to formulate a dosage range that is not toxic for human use. The dosage of the beta-hairpin peptidomimetics of the invention is preferably within a range of circulating concentrations that includes an effective dose with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be selected by The individual physician according to The condition of The patient (see, e.g., Fingl et al, 1975: The pharmacological basis of Therapeutics, Ch.1, p.1).
The following examples illustrate the invention in more detail without intending to limit the scope thereof in any way. The following abbreviations are used in these examples:
HBTU: 1-benzotriazol-1-yl-tetramethyluronium hexafluorophosphate (Knorr et al, Tetrahedron Lett.1989, 30, 1927-;
HCTU: 1-benzotriazole 1- [ bis (dimethylamino) methylene ] -5 chloro-hexafluorophosphate-1, 3-oxide
HOBt: 1-hydroxybenzotriazole;
DIEA: diisopropylethylamine;
HOAT: 7-aza-1-hydroxybenzotriazole;
HATU: o- (7-aza-benzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (Carpino et al, Tetrahedron Lett.1994, 35, 2279-.
Examples
1. Peptide synthesis
Coupling of the first protected amino acid residue to the resin
0.5g of 2-chlorotrityl chloride resin (Barlos et al, Tetrahedron Lett., 1989, 30, 3943-. Suspending the resin in CH2Cl2(2.5 ml) and swollen at room temperature for 30min with constant stirring. Amino acid residue first suitably protected with 0.415mMol (1 eq) (see below) and CH2Cl2The resin was treated with 284. mu.l (4 eq) of Diisopropylethylamine (DIEA) in (2.5 ml) and the mixture was shaken for 4 hours at 25 ℃. Oscillating the resin (CH)2Cl2MeOH/DIEA: 17/2/1), 30ml, 30 min; followed by CH in the following order2Cl2（1x），DMF（1x），CH2Cl2（1x），MeOH（1x），CH2Cl2（1x），MeOH（1x），CH2Cl2（2x），Et2Washed with O (2 ×) and dried under vacuum for 6 hours.
The loading is generally from 0.6 to 0.7 mMol/g.
The following pre-loaded resins were prepared: Fmoc-Pro-2-chlorotrityl resin.
Synthesis of fully protected peptide fragments
The synthesis was performed using a Syro-peptide synthesizer (Multisyntech) using 24 to 96 reaction vessels. In each container 60mg (weight of resin before loading) of the above resin was placed. The following reaction cycles were programmed and performed:
steps 3-6 were repeated to add each amino acid.
The synthesis of fully protected peptide fragments has been terminated, followed by cleavage, cyclization and establishment procedures as described below for peptide preparation.
The analysis method comprises the following steps:
the method comprises the following steps: analytical HPLC retention times (RT in minutes) were determined using a Jupiter protein column (90A, 150X 2.0mm, cod.00F4396-B0-Phenomenex) using the following solvent A (H, in minutes)2O +0.1% TFA) and B (CH)3CN +0.1% TFA), and gradient: 0 min: 95% of A, 5% of B; 20 min: 40% A60% B; 21-23 min: 0% a, 100% B; 23.1-30 min: 95% of A and 5% of B.
The method 2 comprises the following steps: analytical HPLC retention times (RT in minutes) were determined using an Aquity UPLC BEH C18 column (1.7 μm, 100X 2.1mm, cod.186002352-Waters) using the following solvent A (H)2O +0.1% TFA) and B (CH)3CN +0.085% TFA), and gradient: 0 min: 95% of A, 5% of B; 0.2 min: 95% A5% B; 4 min: 35% a, 65% B; 4.2 min: 5% A, 95% B; 4.25 min: 95% of A, 5% of B; 4.9 min: 95% of A and 5% of B.
The procedure is as follows: cleavage, cyclization and establishment of backbone cyclized peptides
Cleavage, backbone cyclization and purification of peptides
After assembly of the linear peptide, the resin was suspended in 1ml (0.39 mMol) of CH2Cl2(v/v) 1% TFA for 3min and filtered, and 1ml (1.17 mMol, 3 eq.) CH2Cl2The filtrate was neutralized with 20% DIEA in (v/v). This procedure was repeated twice to ensure completion of the split. With 2ml CH2Cl2The resin was washed. Will CH2Cl2The layer was evaporated to dryness.
The fully protected linear peptide was dissolved in 8ml dry DMF. 2eq.HATU in dry DMF (1 ml) and 4eq.DIPEA in dry DMF (1 ml) were then added to the peptide followed by stirring for 16 h. The volatiles were evaporated to dryness. The crude cyclic peptide was dissolved in 7ml CH2Cl2Neutralized and extracted three times with 10% acetonitrile in water (4.5 ml). Will CH2Cl2The layer was evaporated to dryness. To completely deprotect the peptide, 3ml of cleavage mixture TFA TIS H was added2O (95: 2.5: 2.5), the mixture was stirred for 2.5 h. The volatiles were evaporated to dryness and the crude peptide was dissolved in 20% AcOH in water (7 ml) and extracted three times with diisopropyl ether (4 ml). The aqueous layer was collected and evaporated to dryness and the residue was purified by preparative reverse phase LC-MS.
After lyophilization, the product was obtained as a white powder and analyzed by HPLC-ESI-MS analysis as described above. Analytical data including purity after preparative HPLC and ESI-MS are shown in table 1.
Examples (Ex.)1-50, shown in table 1. The peptide was initially synthesized with the amino acid L-Pro grafted onto the resin. The starting resin was Fmoc-Pro-2-chlorotrityl resin, which was prepared as described above. The linear peptide was synthesized on the solid support according to the procedure described above in the following order: resin-Pro-DPro-P12-P11-P10-P9-P8-P7-P6-P5-P4-P3-P2-P1. Ex.1-50, cleaved from resin, cyclized, deprotected and purified as shown by preparative reverse phase LC-MS.
After lyophilization, the product was obtained as a white powder and analyzed by HPLC-ESI-MS as described above.
HPLC-retention time (min) was determined using the analytical method described above. Examples 1 to 39 were analyzed using method 1, and for examples 40-50, method 2 was used:
Ex.1(8.87),Ex.2(9.26),Ex.3(9.34),Ex.4(9.45),Ex.5(9.48),Ex.6(9.44),Ex.7(10.11),Ex.8(9.99),Ex.9(10.22),Ex.10(9.76),Ex.11(10.56),Ex.12(11.37),Ex.13(9.13),Ex.14(9.34),Ex.15(8.80),Ex.16(9.23);Ex.17(9.65),Ex.18(9.18),Ex.19(8.37),Ex.20(8.86),Ex.21(8.78),Ex.22(9.32),Ex.23(9.58),Ex.24(9.27),Ex.25(9.31),Ex.26(9.24),Ex.27(9.23),Ex.28(9.34),Ex.29(9.66),Ex.30(9.88),Ex.31(9.62),Ex.32(8.86),Ex.33(9.73),Ex.34(10.46),Ex.35(9.21),Ex.36(9.80),Ex.37(9.73),Ex.38(9.20),Ex.39(9.53),Ex.40(2.07),Ex.41(1.77),Ex.42(1.66),Ex.43(1.67),Ex.44(1.81),Ex.45(1.87),Ex.46(1.81),Ex.47(1.83),Ex.48(1.79),Ex.49(1.88),Ex.50(2.17)。
2. biological method
2.1. Preparation of peptide samples
The lyophilized peptide was weighed on a microbalance (Mettler MT5) and dissolved in sterile water to a final concentration of 1mg/ml unless otherwise indicated. The stock solution was stored at +4 ℃ protected from light.
2.2. Antimicrobial activity of peptides
The selective antimicrobial activity of the peptides was determined in 96-well plates (Nunclon polystyrene) by standard NCCLS broth microdilution methods (see reference 1 below) (slightly modified). Inocula of the microorganisms were diluted in Mueller-Hinton (MH) broth +0.02% BSA and labeled with 0.5McfarlandQuasi-phase comparison to obtain about 106Individual Colony Forming Units (CFU)/ml. Aliquots of the inoculum (50. mu.l) were added to 50. mu.l MH broth containing peptides in serial double dilutions +0.02% BSA. The following microorganisms were used to determine the antibiotic activity of the peptides: escherichia coli (ATCC 25922), Pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC27853, P8191900, P1021903, P1021913, IMP1 Livermore 3140). The antimicrobial activity of the peptides was expressed as the Minimum Inhibitory Concentration (MIC) in μ g/ml, at which no visible growth was observed after incubation at 37 ℃ for 18-20 hours.
2.3. Cytotoxicity test
Cytotoxicity of peptides against HELA cells (Acc 57) and COS-7 (CRL-1651) was determined using MTT reduction assay [ see references 2 and 3 below]. Briefly, the method is as follows: HELA cells and COS-7 cells were cultured at 7.0X 103And 4.5X 103Individual cells/well were seeded and plated in 96-well microtiter plates at 37 ℃ with 5% CO2And culturing for 24 hours. In this regard, the time zero (Tz) was determined by MTT reduction (see below). The supernatant of the remaining wells was discarded and fresh medium and serial dilutions of 12.5, 25 and 50 μ M peptide were dispensed into the wells. Each peptide concentration was determined in triplicate. Culturing the cells at 37 ℃ with 5% CO2The lower time lasts 48 hours. The wells were then washed once with Phosphate Buffered Saline (PBS) and then 100. mu.l of MTT reagent (0.5 mg/ml in media RPMI1640 and DMEM, respectively) was added to the wells. This was incubated at 37 ℃ for 2 hours, followed by aspiration of the medium and addition of 100. mu.l of isopropanol to each well. Measurement of absorbance (OD) of the lysate at 595nm595Peptides). The average value of each concentration was calculated from experiments repeated three times. The percentage growth was calculated as follows: (OD)595peptide-OD595Tz-OD595void)/(OD595Tz-OD595Empty wells) x 100% and curves were made for each peptide concentration.
Using the EXCEL (Microsoft Office 2000) TREND line functions at concentrations (50, 25, 12.5 and 0 μ M), the corresponding growth percentages and values-50, (= TREND (C50:c0,% 50:%0, -50)) determined the LC50 value (lethal concentration, defined as the concentration that kills 50% of the cells) for each peptide. The TREND line functions of the concentrations (50, 25, 12.5 and 0 μ g/ml), the corresponding percentages and the value 50 (= TREND (C) were used50:C0，%50:%050)) the GI50 (growth inhibition) concentration was calculated for each peptide.
2.4. Hemolysis of blood
The peptides were tested for hemolytic activity on human blood red blood cells (hRBC). Fresh hRBCs were washed three times with Phosphate Buffered Saline (PBS) by centrifugation at 2000 Xg for 10 min. Peptides were incubated with 20% v/v hRBC at a concentration of 100. mu.M for 1 hour at 37 ℃. The final concentration of red blood cells is about 0.9X 109Cells/ml. By separately mixing in PBS and H2hRBC were cultured in the presence of 0.1% TritonX-100 in O to determine 0% and 100% lysis values, respectively. The samples were centrifuged and the supernatant diluted 20-fold in PBS buffer and the Optical Density (OD) of the samples at 540nM was measured. 100% lysis value (OD)540H2O) to obtain D of about 1.3 to 1.8540. The% hemolysis was calculated as follows: (OD)540peptide/OD540H2O)×100%。
2.5. Plasma stability
Mu.l of plasma/albumin solution was placed in a polypropylene (PP) tube and spiked (spiked) with 45. mu.l of compound from 100mM solution B derived from 135. mu.l PBS and 15. mu.l 1mM peptide in PBS pH 7.4. A150. mu.l aliquot was transferred to a single well of a10 kDa filter plate (Millipore MAPPB1010 Biomax membrane). For the "0 min control": mu.l PBS was placed in PP tubes and 30. mu.l stock B was added and mixed. Mu.l of control solution was placed in one well of the first plate and served as "filtered control".
In addition, 150 μ l of control solution was placed directly into the storage wells (storage permeate) and served as "non-filtered control". The entire plate including the evaporation lid was incubated at 37 ℃ for 60 min. Plasma samples (rat plasma: Harlan Sera lab UK, human plasma: Blutspendeszentrum Surich) were spun at 4300rpm(3500g) And centrifuged at 15 ℃ for at least 2h to produce 100. mu.l of filtrate. For "serum albumin" -samples (freshly prepared human albumin: Sigma A-4327, rat albumin: Sigma A-6272, all at a concentration of 40mg/ml in PBS), centrifugation for about 1 hour was sufficient. The permeate from the stored PP plates was analyzed by LC/MS as follows: column: jupiter C18 (Phenomenex), mobile phase: (A) 0.1% formic acid and (B) acetonitrile in water, gradient: (B) 5% -100% in 2min, electrospray ionization, MRM detection (triple quadrupole). The peak areas were determined and the average of the triplicate values was calculated. By: 100- (100 XT)60/T0) Binding was expressed as a percentage of controls 1 and 2 (filtered and non-filtered time points 0 min). The average of these values was calculated.
2.6 pharmacokinetic Studies (PK)
Pharmacokinetic studies in mice after a single intravenous, subcutaneous and intraperitoneal administration
Pharmacokinetic studies of the compound of example 1 ("ex.1") after a single intravenous (i.v.) and subcutaneous (s.c.) administration. CD-1 mice (20-25 g) were used in the study. Physiological saline was used as the carrier. The volume was 2ml/kgi.v., 5ml/kg s.c. and peptide ex.1 was injected to obtain a final intravenous dose of 1mg/kg and a subcutaneous dose of 5 mg/kg. Approximately 200-. Plasma was removed from the pelleted cells after centrifugation and frozen at-80 ℃ prior to HPLC-MS analysis.
Preparation of plasma calibration samples
"blank" mouse plasma from untreated animals was used. 0.2ml aliquots of each plasma were spiked with 50ng of propranolol (InternalStandard, IS) (by mixing inSamples were prepared by solid phase extraction on HLB cartridges (Waters) and known amounts of ex.1 were used in order to obtain 9 calibration samples in the range 10-5000 nM. With 1ml of methanol and then with 1% NH in 1ml of water3To adjustHLB cartridges. Then with 700. mu.l of 1% NH in water3The samples were diluted and loaded. 1ml of methanol/1% NH in 5/95 water3To wash the plate. Elution was performed using 0.1% TFA in 1ml methanol.
The plate containing the eluate was introduced into a concentration system and allowed to dry. The residue was dissolved in 100. mu.L formic acid 0.1%/acetonitrile 95/5 (v/v) and analyzed in HPLC/MS on a reverse phase analytical column (Jupiter C18, 50X 2.0mm, 5 μm, Phenomenex) using gradient elution (mobile phase A: 0.1% formic acid in water, B: acetonitrile; from 5% B to 100% B in 2 min).
Preparation of plasma samples
Samples from animal treatments were pooled in order to obtain the appropriate volume for extraction. If a total volume of less than 0.2ml is obtained, appropriate amounts of "blank" mouse plasma are added to maintain consistency with the matrix of the calibration curve. The samples were then spiked with IS and processed as described by the calibration curve.
Pharmacokinetic evaluation
The consolidated data (typically n =2 or 3) were subjected to PK analysis using software Win Nonlin (Pharsight). The area under the curve AUC was calculated by the linear trapezoidal rule. The elimination half-life was calculated by linear regression for at least three data points during the elimination phase. By a correlation coefficient (r)2) The time interval chosen for the half-life determination is evaluated and should at least exceed 0.85 and preferably exceed 0.96. In the case of i.v. administration, the starting concentration of t zero was determined by extrapolating the curves for the first two time points. Finally, after s.c. to i.v. administration, the bioavailability after i.p. administration was quantitatively calculated from the standardized AUCinf _ D _ obs.
2.7. In vivo sepsis testing
Male mice derived from the 6CD-1 (Crl.) group weighing 24. + -.2 g were used. Mice were each treated with LD90-100 Pseudomonas aeruginosa (ATCC 27853) (9X 10) in brain-heart infusion without 5% mucin6CFU/0.5 ml/mouse) Intravenous (IV) vaccination. The test animals were dosed Subcutaneously (SC) with 5, 2.5, 1, 0.5, 0.25 and 0.1mg/kg of compound, vehicle (0.9% NaCl, 10 ml/kg) 1 hour after inoculation of the bacteria. In addition, at 1 and 6 hours after bacterial inoculation, another group was treated twice with a dose of 5mg/kg of compound. Mortality was recorded once daily for 7 days after bacterial inoculation, and a 50% or higher (350%) increase in survival relative to vehicle control animals after bacterial inoculation was indicative of significant antimicrobial effect. MED (ED 50) was determined by non-linear regression using Graph-Pad Prism (Graph Pad Software, USA).
2.8. Results
The results of the experiments described in 2.2, 2.3 and 2.4 above are shown in table 2 below.
The experimental results described in 2.5 above are shown in table 3 below.
TABLE 3
Examples Stable human plasma t1/2（min） Stable rat plasma t1/2（min）
1 300 300
2 300 300
6 300 300
16 300 300
22 300 300
24 300 300
25 300 300
28 300 300
29 300 300
32 300 300
35 300 300
The results of the experiment described in 2.6 (PK) above are shown in table 4 below.
TABLE 4
Ex.1 was followed by intravenous kinetics for Ex.1 after intravenous administration of 1mg/kg body weight. After PK analysis, Ex.1 showed a putative C of 2174ng/mlInitially, the process is startedAnd 1268ng/ml of C observed at 5minmax. Plasma levels rapidly dropped to 575 and 177ng/ml at 15min and 1 hour, respectively. From 0.5 to 2h, with a clearance half-life of 0.53h, plasma levels dropped to 10.6ng/ml at 3 h. AUCINF _ obs reaches 679.5 ng.h/ml.
After subcutaneous administration of Ex.1 at a dose level of 5mg/kg body weight, the plasma level of Ex.1 increased between 0.5 and 1h and showed a C of 2333ng/mlmax. From 0.5 to 8h, with a clearance half-life of 0.95h, the plasma levels dropped to 7.3ng/ml at 8 h. AUCINF _ obs reaches 4016.5 ng.h/ml.
Compared to the normalized AUC values after i.v. administration (100% absorbed, 679 ng.h/ml) ex.1, the absorption after s.c. administration reached 118% (803 ng.h/ml). Values in excess of 100% may partially reflect impaired reliability caused by a limited number of points or by non-linear dose.
The experimental results (sepsis test) described in 2.7 above are shown in tables 5 to 7 below.
Sepsis assay in mice: LD90-100 Pseudomonas aeruginosa (ATCC 27853) (9X 10)6CFU/0.5 ml/mouse IV) and is after 1h ex.1s.c.
TABLE 5
Sepsis assay in mice: LD90-100 Pseudomonas aeruginosa (ATCC 27853) (9X 10)6CFU/0.5 ml/mouse IV) and after 1 and 5h ex.40s.c.
TABLE 6
Sepsis assay in mice: LD90-100 Pseudomonas aeruginosa (ATCC 27853) (9X 10)6CFU/0.5 ml/mouse IV) and was after 1 and 5h ex.50s.c. of.
TABLE 7
Reference to the literature
1. The national committee for clinical laboratory standards, 1993. Dilute antimicrobial susceptibility testing method for aerobic growing bacteria, third edition, approved Standard M7-A3. Clinical laboratory standard national committee, Villanova, Pa.
2.Mossman T.J Immunol Meth 1983，65，55-63
3.Berridge MV，Tan AS.Archives of Biochemistry&Biophysics1993，303，474-482

Claims (13)

1. A compound of the formula
Wherein
Is thatDPro-LPro, and
z is a chain of 12 α -amino acid residues, the positions of which in the chain are counted starting from the N-terminal amino acid, whereby these amino acid residues at positions P1-P12 in the chain are:
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Gly; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11:DAla; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11:DVal; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Aib; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Abu; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Leu; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ile; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Nle; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Nva; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Chg; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Cha; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Gln; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11 Asp; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Glu; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Thr; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Pro; and
-P12:Dab；
or
-P1:Cit；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Tyr；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Tyr；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:His；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Ala；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Ala；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Dab；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Val; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Lys；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Lys；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Lys；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Lys；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Lys；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Gln；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Gln；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Val; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Nle；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Val；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Chg；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Cha；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
Hse, P11; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11 t-BuG; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Cpa; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Asn; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:Dab；
-P7:Gln；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Gln；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Gln；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Asp；
-P2:Tyr；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Asp；
-P2:Tyr；
-P3:Ile；
-P4:Dab；
-P5:Orn；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Glu；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Glu；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Gln；
or
-P1:Glu；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Lys；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Glu；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Orn；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Thr；
-P2:Trp；
-P3:Ile；
-P4:Dab；
-P5:Orn；
-P6:DDab；
-P7:Dab；
-P8:Trp；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Dab；
or
-P1:Glu；
-P2:Trp；
-P3:Ile；
-P4:Gln；
-P5:Lys；
-P6:Dab；
-P7:Dab；
-P8:Ser；
-P9:Dab；
-P10:Dab；
-P11: Ala; and
-P12:Ser；
and pharmaceutically acceptable salts thereof.
2. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically inert carrier.
3. A composition according to claim 2, in a form suitable for oral, topical, or injectable administration.
4. A composition according to claim 2, in a form suitable for transmucosal administration.
5. A composition according to claim 2, in a form suitable for transdermal, buccal or pulmonary administration.
6. A composition according to claim 2 in the form of a tablet, lozenge, capsule, liquid, gel, cream, ointment, spray or suppository.
7. A composition according to claim 2, in the form of a solution.
8. A composition according to claim 2, in the form of a suspension.
9. A composition according to claim 2, in the form of a syrup.
10. Use of a compound according to claim 1 for the preparation of a medicament for the treatment of an infection caused by pseudomonas aeruginosa or a disease associated with such an infection.
11. Use of a compound according to claim 1 as a fungicide or preservative for food and cosmetics.
12. A process for the preparation of a compound according to claim 1, which process comprises
(a) Coupling an appropriately functionalized solid support with an appropriate N-protected derivative of an amino acid at position 5, 6 or 7 in the desired end product, any functional groups that may be present in said N-protected amino acid derivative likewise being appropriately protected;
(b) removing the N-protecting group from the product thus obtained;
(c) coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position in the desired end product close to the N-terminal amino acid residue, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(d) removing the N-protecting group from the product thus obtained;
(e) repeating steps (c) and (d) until the N-terminal amino acid residue has been introduced;
(f) coupling the product thus obtained with a compound of the general formula
Wherein
The definitions are the same as defined in claim 1, and X is an N-protecting group, or, alternatively,
(fa) reacting the product obtained in step (e) withLAppropriate N-protected derivatives of Pro;
(fb) removing the N-protecting group from the product thus obtained; and
(fc) reaction of the product thus obtained withDAppropriate N-protected derivatives of Pro;
(g) removing the N-protecting group from the product obtained in step (f) or (fc);
(h) coupling the product thus obtained with a suitable N-protected derivative of the amino acid at position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative likewise being suitably protected;
(i) removing the N-protecting group from the product thus obtained;
(j) coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position remote from position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative being likewise suitably protected;
(k) removing the N-protecting group from the product thus obtained;
(l) Repeating steps (j) and (k) until all amino acid residues have been introduced;
(o) separating the product thus obtained from the solid support;
(p) cyclizing the product isolated from the solid support; and
(s) if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable or unacceptable salt thus obtained into the corresponding free compound of the general formula I or into a different pharmaceutically acceptable salt.
13. A process for the preparation of a compound according to claim 1, which process comprises
(a') coupling an appropriately functionalized solid support with a compound of the general formula
Wherein
The definition is the same as defined in claim 1, and X is an N-protecting group, or, alternatively
(a' a) contacting said suitably functionalized solid support withLAppropriate N-protected derivatives of Pro;
(a' b) removing the N-protecting group from the product thus obtained; and
(a' c) reacting the product thus obtained withDAppropriate N-protected derivatives of Pro;
(b ') removing the N-protecting group from the product obtained in step (a ') or (a ' c);
(c') coupling the product thus obtained with a suitable N-protected derivative of the amino acid at position 12 in the desired end product, any functional groups which may be present in said N-protected amino acid derivative likewise being suitably protected;
(d') removing the N-protecting group from the product thus obtained;
(e') coupling the product thus obtained with a suitable N-protected derivative of an amino acid at a position remote from position 12 in the desired end product, any functional group which may be present in said N-protected amino acid derivative being likewise suitably protected;
(f') removing the N-protecting group from the product thus obtained;
(g ') repeating steps (e ') and (f ') until all amino acid residues have been introduced;
(i') separating the product thus obtained from the solid support;
(j') cyclizing the product isolated from the solid support; and
(m') if desired, converting the product thus obtained into a pharmaceutically acceptable salt or converting a pharmaceutically acceptable or unacceptable salt thus obtained into the corresponding free compound of the general formula I or into a different pharmaceutically acceptable salt.
HK13102561.0A 2006-01-16 2013-02-28 Template-fixed peptidomimetics with antimicrobial activity HK1175478B (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PCT/CH2006/000036 WO2007079597A1 (en) 2006-01-16 2006-01-16 Template - fixed peptidomimetics with antimicrobial activity
CHPCT/CH2006/000036 2006-01-16

Publications (2)

Publication Number Publication Date
HK1175478A1 HK1175478A1 (en) 2013-07-05
HK1175478B true HK1175478B (en) 2016-03-18

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