+

WO1999028339A1 - Fragments de peptides cycliques de squelette et leurs analogues derives de pban 1-33, utiles en tant qu'insecticides - Google Patents

Fragments de peptides cycliques de squelette et leurs analogues derives de pban 1-33, utiles en tant qu'insecticides Download PDF

Info

Publication number
WO1999028339A1
WO1999028339A1 PCT/IB1998/001924 IB9801924W WO9928339A1 WO 1999028339 A1 WO1999028339 A1 WO 1999028339A1 IB 9801924 W IB9801924 W IB 9801924W WO 9928339 A1 WO9928339 A1 WO 9928339A1
Authority
WO
WIPO (PCT)
Prior art keywords
backbone
pban
peptides
insect
peptide fragments
Prior art date
Application number
PCT/IB1998/001924
Other languages
English (en)
Inventor
Chaim Gilon
Miriam Altstein
Original Assignee
Yissum Research Development Company Of The Hebrew University Of Jerusalem
State Of Israel/Ministry Of Agriculture
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Yissum Research Development Company Of The Hebrew University Of Jerusalem, State Of Israel/Ministry Of Agriculture filed Critical Yissum Research Development Company Of The Hebrew University Of Jerusalem
Priority to AU12542/99A priority Critical patent/AU1254299A/en
Publication of WO1999028339A1 publication Critical patent/WO1999028339A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention generally relates to backbone cyclic peptides consisting of fragments derived from the amino acid sequence of Pheromone Biosynthesis Activating Neuropeptide (hereinafter referred to as PBAN 1-33, PBAN (1-33)NH 2 or PBAN) and their analogs that have been found to be useful as insecticides, and to conformational libraries thereof, that have been found to be useful as a basis for the design of improved antagonist insecticides and for the discovery of insecticides.
  • the present invention further relates to a method for controlling insects, preferably moths, utilizing said backbone cyclic peptide fragments and analogs derived from the PBAN sequence.
  • Compounds used as insecticides should be of low molecular weight, have high penetrability through the insect cuticle, and be environmentally stable. This is particularly essential for those compounds that are meant to be applied to adult insects as contact insecticides.
  • the present invention relates to a novel group of environmentally friendly insecticides, namely neuropeptide antagonists and their analogs.
  • Neuropeptide antagonists may be either of endogenous origin (naturally occurring) or synthetic compounds. Such antagonists may either be derived from, or resemble, the natural peptides, and be peptidic or of a peptidomimetic nature.
  • the term peptidomimetic describes molecules that are not completely peptidic in nature, such as pseudo-peptides and semi-peptides (G.J. Moore (1994) ZZPS 15:124-129). There are clear advantages for using a mimetic of a given peptide rather than linear peptides.
  • peptidomimetics are of low molecular weight, stable, highly penetrable, and cheaper to produce than peptides.
  • a conformational model must be selected in which the biologically active groups can be arranged in a manner that allows them to retain the spatial relationship that existed in the biologically active parent peptide.
  • Cyclic peptides on the other hand, have a constrained conformation and can serve, therefore, as the basis for the determination of bioactive conformation, that can be further used for the design of peptidomimetic insecticides (G. L. Moore (1994) ⁇ PS 15:124- 129.
  • Cychc peptides also usually possess enhanced bioavailabihty, due to increased metabolic stability. The enhanced stability could allow diminished doses of insecticides at longer intervals and the restricted conformation would improve the insecticide selectivity.
  • Backbone cychc peptides which have the inherent advantages of cyclic peptides, are characterized in that they incorporate building units with modified side chains attached to the nitrogens of their amino acids. These building units permit the synthesis of backbone cychc peptidomimetics (Gilon et al. (1991) Biopolymers, 31:745).
  • Backbone cychc peptide libraries have been applied for generating mixtures of p tidomimetic compounds (such as in WO 97/09344 (Gilon et al.)).
  • the method of obtaining backbone cyclic peptides enables cyclization of a given peptide sequence without major perturbation of biologically essential moieties, such as side chains, and amino and carboxyl terminal groups.
  • optimization of the peptide conformation can be achieved by allowing permutation of the mode of cyclization, ring location along the peptide sequence, ring size and ring chemistry.
  • Screening backbone cyclic peptide libraries (Cycloscan) allows fast discovery of peptides with optimal conformation for the performance of its biological role.
  • PBAN Pheromone Biosynthesis Activating Neuropeptide
  • the present invention relates to backbone cychc peptide fragments or analogs derived from the amino acid sequence of PBAN. (1-33)NH 2 for use as insecticides.
  • the present invention further relates to conformational libraries based on backbone cychc peptide fragments or analogs derived from the amino acid sequence of PBAN (1-33)NH 2 for use as a basis for the design of PBAN antagonists and for the discovery of insecticides.
  • the present invention therefore, discloses significantly superior peptides for use as insecticides, and conformational libraries thereof useful as a basis for the design of PBAN antagonists and the discovery of insecticides.
  • the present invention relates to backbone cyclic peptides or their analogs derived from the amino acid sequence of PBAN (1-33)NH 2 , wherein the peptides are fragments or analogs thereof of PBAN (1-33)NH 2 , useful as insecticides.
  • Said backbone cyclic peptides have the general Formula(I):
  • z and y each independently are an integer from 1-10;
  • AA designates a naturally occurring or synthetic amino acid residue;
  • E represents a functional group such as OH, NH 2 etc., N-CH(R)- designates a building unit, wherein R is H or an amino acid side chain, and the lines above the formula (1) designate a bridge and spacer of the Formula ⁇ .
  • the peptides of the present invention are modified C-terminal fragments of PBAN (1-33)NH 2 with agonistic activity, of the formula: Arg-Tyr-Phe-Ser-Pro-Arg-Leu-NH 2 , or with antagonistic activity of the formula: Arg-Tyr-Phe-(D)-Phe-Pro-Arg-Leu-NH 2 .
  • Replacing the Pro residue, in both cases, is an N(amino-alkyl)Gly or other building unit having various length of the alkyl chain. This building unit is cyclized through its amine function via a lactam bridge to a dicarboxylic acid spacer attached to the N terminus.
  • the present invention further relates to backbone cychc conformational libraries, preferably soluble, based on said peptides or analogs, useful as a basis for the design and discovery of neuropeptide based antagonist insecticides.
  • the insecticidal activity of the said backbone cychc antagonistic peptides or analogs is aimed at the disruption of pheromone production, melanization and other key neuroendocrine physiological functions in insects, controlled by PBAN (1-33)NH 2 and derived fragments or analogs thereof.
  • the said backbone cychc peptides or analogs are useful for moth control, more specifically for Heliothis peltigera and Spodoptera littioralis moth species control.
  • the present invention further relates to a method for insect control comprising applying the said backbone cyclic peptides on insects,
  • the present invention relates to backbone cyclic peptide fragments and their analogs derived from Pheromone Biosynthesis Activating Neuropeptide (PBAN (1-33)NH 2 ), useful as insecticides and to conformational libraries thereof, useful as a basis for the design and discovery of insecticides, specifically of a new environmentally friendly group of neuropeptide based antagonist insecticides aimed at the disruption of key neuroendocrine physiological functions in insects.
  • the present invention further relates to a method for controlling insects, particularly moths, utilizing said backbone cychc peptide fragments and analogs derived from the PBAN (1-33)NH 2 .
  • the use of these peptides disrupts mating in moths by inhibiting sex pheromone biosynthesis and inhibits cuticular melanization, and other activities in insects.
  • Soluble conformational libraries based on backbone cyclic peptides derived from PBAN were prepared and screened in order to find a potent PBAN antagonist, which fulfills the characteristics required from a neuropeptide based antagonist insecticide.
  • the strategy of synthesizing backbone cyclic peptides was employed with the distinct benefits of: a) achieving a metabolically stable PBAN antagonist and b) the possibility of achieving high penetrability of the PBAN antagonist.
  • the backbone cyclic peptides described in the present invention have the general Formula (1):
  • two sub-libraries of backbone cyclic analogs were designed and synthesized.
  • One conformational sub-library (cyclic Ser library - Fig. IA) was based on a slight modification of the sequence of the C-terminal fragment of PBAN (MINI-PBAN) (Tyr-Phe-Ser-Pro-Arg-Leu-NH2), which was shown to be as potent as the frill length PBAN.
  • the second backbone cyclic sub- library (cyclic D-Phe library - Fig, IB) was based on the sequence of the most potent linear antagonist (Arg-Tyr-Phe-(D)-Phe-Pro-Arg-Leu-NH2) that was found.
  • Cycloscan technology For the preliminary screening, all the cyclic peptides in each sub- hbrary had the same sequence and the same location of the ring. The members of each library differed from each other by the ring size and the chemistry of the ring. The general structure of the two sub-libraries is shown in the following A & B, (CH 2 ) m -CO-NH--(CH 2 ) n .
  • (B) sub-libraries of peptides For the first generation of the sub-libraries, the Pro residue in the Ser and the D-Phe peptides, was replaced by N(amino-alkyl)Gly building unit having various length of the alkyl chain. The unit was cychzed through its amine function via a lactam bridge to a dicarboxylic acid spacer attached to the N-terminus. In addition to differences in ring size between members of each sub-library, peptides having the same ring size but differing from each other by the location of the lactam bridge, were synthesized.
  • the peptides in the two sub-libraries were synthesized by the Simultaneous Multiple Peptide Synthesis methodology (SMPS) on Rink Amide MBHA Resin (loading 0.55 mmol gm) using Fmoc chemistry.
  • SMPS Simultaneous Multiple Peptide Synthesis methodology
  • the following protected amino acids were used: Fmoc-LeuOH, Fmoc-Arg(Pmc)OH, FmocSer(t-Bu)OH Fmoc-PheOH, Fmoc-(D)PheOH Fmoc-Tyr(t-Bu)OH.
  • the following backbone cyclic building units were used: Boc- NH(CH 2 ) n -N(Fmoc)CH 2 -COOH ( ⁇ F 2, 3, 4, 6).
  • Coupling was performed by preactivated solution of three fold excess of Fmoc amino acids or building units with TBTU for 5 min. in NMP.
  • Deprotection of Boc from the amino alkyl group of the building units was carried out with Cl j SiF, prepared in situ from SiCl 4 and NaF in ACN:DCM (1: 1).
  • One resin cyclization was achieved by 6 fold excess of TBTU and repeated 4 times.
  • the peptides were removed from the resin with concomitant side chain deprotection by 90% TFA with addition of scavengers (H 2 0:ethanedithiol:thioanisole 2.5%; 2.5%: 5%).
  • the crude peptides were purified on C8 prep HPLC using the ACN:H 2 0 gradient.
  • the data in Table 3 depict the presence of 5 antagonistic peptides (No. 20, 21, 22, 25, 28) exhibiting inhibitory activities ranging from 70% to 79%. Due to possible variations among moth colonies, and in order to determine the reproducibility of the results, the inhibitory activity of the D-Phe backbone cychc peptides was tested on two other moth colonies. As can be seen from Table 4, some variability among moth colonies does exist. Three out of the five peptides (20, 21 and 25) exhibited antagonistic activity in all colonies. Peptides 22 and 28 were active only in two out of the three colonies.
  • Antagonistic activity was determined in H. peltigera females by injection of each one of the peptides (at a concentration of 1000 pmol) together with 0.5 or 1 pmol PBAN 1- 33NH 2 .
  • the antagonistic activity is expressed as 100 minus the ratio (in %) between the pheromone content evoked by the injection of PBAN 1-33NH 2 in the presence and absence of each one of the peptides.
  • the amount of pheromone evoked by 0. 5 or 1 pmol PBAN 1- 33NH 2 ranged from 50-160 ng.
  • Pheromone content in each treatment was monitored with at least 10 females.
  • Table 4 Comparison of the pheromonotropic antagonistic activity of some of the backbone cyclic peptides of the D-Phe sub-library in different moth colonies
  • antagonistic compounds may also exhibit full or partial agonistic activity
  • the antagonistic peptides were tested for their ability to evoke pheromone biosynthesis in the absence of PBAN 1-33NH,.
  • Agonistic activity was performed using the above mentioned pheromonotropic in vivo bioassay. Peptides were injected at a concentration of 1000 pmol and pheromone content was momtored 2 hours post injection by CGC. As can be seen from Table 5, four out of the five antagonists were devoid of agonistic activity, Only one (peptide 21) was agonistic.
  • Agonistic activity was determined in Hpeltigera females by injection of each one of the peptides at a concentration of 1000 pmol.
  • the agonistic activity is expressed as the ratio (in %) between the pheromone content evoked by the injection of each one of the cyclic peptides, and PBAN 1-33NH, (at the same concentration). All other details are as described in the legend to Table 3.
  • the peptides of the Ser sub-library that did not exhibit, agonistic activity were tested for their ability to act as antagonists, namely to inhibit sex pheromone biosynthesis evoked by exogenously administered PBAN1-33NH 2 .
  • the study was performed using the pheromonotropic in vivo bioassay described above. Pheromone biosynthesis was evoked by 0.5 or 1 pmol PBAN.1-33NH 2 .
  • Each peptide from the Ser sub-library (4, 5, 7-9, 11-13, Table 1) was tested at a concentration of 1000 pmol. Peptides were injected together with PBAN1-33NH,, and pheromone content in the pheromone gland was monitored 2 hours post injection by CGC. The data in Table 7 show that none of the tested peptides exhibited antagonistic activity.
  • Table 8 depict the antagonistic activity of only one peptide 28. This peptide inhibited the PBAN evoked melanization to a level that did not differ significantly from that of ligated larvae injected only with buffer. All other peptides, including those that showed low melanization content (e.g. 21, 24, 25 and 26) did not differ significantly from PBAN injected larvae.
  • Table 8 -Melanotropic antagonistic activity of the D-Phe sub-library of backbone cyclic peptides

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Insects & Arthropods (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

Peptides cycliques de squelette consistant en des fragments dérivés de la séquence aminoacide du neuropeptide d'activation de biosynthèse de phéromones (PBAN) et leurs analogues. Ces peptides sont utiles en tant qu'insecticides. De plus, l'invention concerne leurs banques de conformation pouvant servir de base de conception d'insecticides antagonistes améliorés et de découverte d'insecticides. Elle concerne également un procédé servant à lutter contre des insectes, de préférence, les lépidoptères noctuidés, au moyen desdits fragments de peptides cycliques de squelette et de leurs analogues dérivés de la séquence de PBAN.
PCT/IB1998/001924 1997-12-02 1998-12-02 Fragments de peptides cycliques de squelette et leurs analogues derives de pban 1-33, utiles en tant qu'insecticides WO1999028339A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU12542/99A AU1254299A (en) 1997-12-02 1998-12-02 Backbone cyclic peptide fragments and their analogs derived from pban 1-33, use ful as insecticides

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US98240597A 1997-12-02 1997-12-02
US08/982,405 1997-12-02

Publications (1)

Publication Number Publication Date
WO1999028339A1 true WO1999028339A1 (fr) 1999-06-10

Family

ID=25529136

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB1998/001924 WO1999028339A1 (fr) 1997-12-02 1998-12-02 Fragments de peptides cycliques de squelette et leurs analogues derives de pban 1-33, utiles en tant qu'insecticides

Country Status (2)

Country Link
AU (1) AU1254299A (fr)
WO (1) WO1999028339A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0564739A2 (fr) * 1991-10-02 1993-10-13 Yissum Research Development Company Of The Hebrew University Of Jerusalem Peptides ayant une chaîne principale cyclique, leur préparation et compositions pharmaceutiques les contenant
WO1995029191A1 (fr) * 1994-04-24 1995-11-02 State Of Israel Fragments de peptides et analogues derives du pban 1-33 nh2 et destines a la lutte contre les mites
WO1997009344A2 (fr) * 1995-08-29 1997-03-13 Peptor Limited Banques de nouveaux analogues de peptides a squelette cyclise sur le squelette

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0564739A2 (fr) * 1991-10-02 1993-10-13 Yissum Research Development Company Of The Hebrew University Of Jerusalem Peptides ayant une chaîne principale cyclique, leur préparation et compositions pharmaceutiques les contenant
US5723575A (en) * 1991-10-02 1998-03-03 Yissum Research Development Company Of The Hebrew University Of Jerusalem Process for the preparation of backbone cyclic peptides
WO1995029191A1 (fr) * 1994-04-24 1995-11-02 State Of Israel Fragments de peptides et analogues derives du pban 1-33 nh2 et destines a la lutte contre les mites
WO1997009344A2 (fr) * 1995-08-29 1997-03-13 Peptor Limited Banques de nouveaux analogues de peptides a squelette cyclise sur le squelette

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BIOLOGICAL ABSTRACTS, 1 January 1900, Philadelphia, PA, US; abstract no. 1998-473558, XP002916820 *
GILON C, ET AL.: "BACKBONE CYCLIZATION: A NEW METHOD FOR CONFERRING CONFORMATIONAL CONSTRAINT ON PEPTIDES", BIOPOLYMERS, JOHN WILEY & SONS, INC., US, vol. 31, 1 January 1991 (1991-01-01), US, pages 745 - 750, XP002916821, ISSN: 0006-3525, DOI: 10.1002/bip.360310619 *

Also Published As

Publication number Publication date
AU1254299A (en) 1999-06-16

Similar Documents

Publication Publication Date Title
Scherkenbeck et al. Insect neuropeptides: structures, chemical modifications and potential for insect control
Fónagy et al. Isolation, primary structure and synthesis of neomyosuppressin, a myoinhibiting neuropeptide from the grey fleshfly, Neobellieria bullata
Altstein Novel insect control agents based on neuropeptide antagonists: The PK/PBAN family as a case study
Altstein et al. Advances in the application of neuropeptides in insect control
DE69225597T2 (de) Peptide mit einer lanthionin-brücke
Nachman et al. Active conformation of the pyrokinin/PBAN neuropeptide family for pheromone biosynthesis in the silkworm
US5959182A (en) Insecticidal toxins derived from funnel web (atrax or hadronyche) spiders
Zhang et al. Synthesis, aphicidal activity and conformation of novel insect kinin analogues as potential eco‐friendly insecticides
Taneja‐Bageshwar et al. Comparative structure‐activity analysis of insect kinin core analogs on recombinant kinin receptors from Southern cattle tick Boophilus microplus (Acari: Ixodidae) and mosquito Aedes aegypti (Diptera: Culicidae)
Nachman et al. A C-terminal aldehyde insect kinin analog enhances inhibition of weight gain and induces significant mortality in Helicoverpa zea larvae
CN101519430A (zh) 一类新型保幼激素合成抑制剂－苯丙-甘-亮三肽酰胺类似物
Zeltser et al. Discovery of a linear lead antagonist to the insect pheromone biosynthesis activating neuropeptide (PBAN)☆
AU5605998A (en) Synthetic antibiotics
SIMMONDS et al. Synthesis of disulfide‐bridged fragments of ω‐conotoxins GVIA and MVIIA: Use of Npys as a protecting/activating group for cysteine in Fmoc syntheses
Nachman et al. Pseudodipeptide analogs of the pyrokinin/PBAN (FXPRLa) insect neuropeptide family containing carbocyclic Pro-mimetic conformational components
Saito et al. The core and complementary sequence responsible for biological activity of the diapause hormone of the silkworm, Bombyx mori
WO1999028339A1 (fr) Fragments de peptides cycliques de squelette et leurs analogues derives de pban 1-33, utiles en tant qu'insecticides
Zeltser et al. Insect neuropeptide antagonist. Part II. Synthesis and biological activity of backbone cyclic and precyclic PBAN antagonists
US7078384B2 (en) Mimetic insect allatostatin analogs for insect control
Puiroux et al. The effect of proctolin analogues and other peptides on locust oviduct muscle contractions
NACHMAN et al. An active pseudopeptide analog of the leucokinin insect neuropeptide family
Nachman et al. Evaluation of a PK/PBAN analog with an (E)-alkene, trans-Pro isostere identifies the Pro orientation for activity in four diverse PK/PBAN bioassays
NACHMANI et al. Acyl, pseudotetra‐, tri‐and dipeptide active‐core analogs of insect neuropeptides
DE69312698T2 (de) Trizyklische tachikinin-antagonisten, deren herstellung und diese enthaltende pharmazeutische präparate
Quartara et al. Influence of lipophilicity on the biological activity of cyclic pseudopeptide NK-2 receptor antagonists

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CZ DE DK EE ES FI GB GE GH GM HR HU ID IL IS JP KE KG KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
NENP Non-entry into the national phase

Ref country code: KR

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载