FR2828686A1 - New peptides containing amino acids present in regions of gp36 protein of a strain of feline immune deficiency virus, useful for treating feline immune deficiency virus infection in cats - Google Patents

New peptides containing amino acids present in regions of gp36 protein of a strain of feline immune deficiency virus, useful for treating feline immune deficiency virus infection in cats Download PDF

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FR2828686A1
FR2828686A1 FR0110910A FR0110910A FR2828686A1 FR 2828686 A1 FR2828686 A1 FR 2828686A1 FR 0110910 A FR0110910 A FR 0110910A FR 0110910 A FR0110910 A FR 0110910A FR 2828686 A1 FR2828686 A1 FR 2828686A1
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deficiency virus
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FR2828686B1 (en
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Pierre Francois Serres
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
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    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
    • C12N2740/15022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

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Abstract

Peptides (I) containing at least 25 amino acids (aa) present in the regions 649-696 or 729-782 of the gp36 protein of a strain of feline immune deficiency virus (FIV) or of the corresponding consensus sequence, but having no more than 40 consecutive aa from the gp36 protein. Peptides (I) containing at least 25 amino acids (aa) present in the regions 649-696 or 729-782 of the gp36 protein of a strain of feline immune deficiency virus (FIV) or of the corresponding consensus sequence, but having no more than 40 consecutive aa from the gp36 protein. In (I), Lys may be replaced by Arg and Ile by Leu, or vice versa.

Description

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La présente invention concerne de nouveaux peptides utilisables comme médicaments permettant de lutter contre l'infection par le virus de l'immunodéficience féline (FIV) chez le chat.  The present invention relates to new peptides which can be used as medicaments for combating infection with the feline immunodeficiency virus (FIV) in cats.

On sait que l'infection par le FIV est une infection répandue et ancienne qui pose d'importants problèmes de santé chez le chat domestique dans le monde entier ; voir par exemple G. Pancino et al., Annales de l'Institut Pasteur 5 (1), 25-34 (1994), et M.  It is known that IVF infection is a widespread and ancient infection which poses serious health problems in domestic cats worldwide; see for example G. Pancino et al., Annales de Institut Pasteur 5 (1), 25-34 (1994), and M.

Bendinelli et al., Clinical Microbiology Reviews, 8 (1), 87-112 (1995). Bendinelli et al., Clinical Microbiology Reviews, 8 (1), 87-112 (1995).

On a maintenant découvert de nouveaux peptides utilisables en thérapeutique vétérinaire pour traiter les infections par le FIV chez le chat.  We have now discovered new peptides which can be used in veterinary therapy to treat IVF infections in cats.

Ces peptides ont la propriété d'inhiber le phénomène de fusion membranaire qui est le processus-clé permettant au virus FIV de pénétrer dans ses cellules cibles (principalement les lymphocytes T).  These peptides have the property of inhibiting the phenomenon of membrane fusion which is the key process allowing the IVF virus to penetrate into its target cells (mainly T lymphocytes).

On sait que dans le traitement du SIDA humain, on utilise actuellement un traitement dit"trithérapie"dont le prix de revient est très élevé, et dont l'efficacité est remise en question par l'apparition de souches virales résistantes.  We know that in the treatment of human AIDS, we currently use a so-called "triple therapy" treatment, the cost price of which is very high, and the effectiveness of which is called into question by the appearance of resistant viral strains.

Il n'est pas possible d'envisager en pratique l'utilisation des trithérapies pour traiter les infections à FIV chez le chat, compte tenu de leur coût prohibitif.  It is not possible to consider in practice the use of triple therapies to treat IVF infections in cats, given their prohibitive cost.

Au contraire, les peptides constituent des principes actifs de faible coût. En outre, les peptides utilisables selon l'invention ont des séquences correspondant à certaines zones de la protéine transmembranaire d'enveloppe du FIV qui sont hautement conservées. Il n'y a donc pratiquement pas de risques d'apparition de souches virales résistantes, car ces zones hautement conservées correspondent à des séquences peptidiques qui sont très utiles, voire indispensables au virus, et dont certains acides aminés-clés ne peuvent en aucun cas être mutés sans compromettre l'entrée du virus, et donc sa réplication, dans les cellules cibles.  On the contrary, peptides constitute active ingredients of low cost. In addition, the peptides which can be used according to the invention have sequences corresponding to certain areas of the transmembrane protein of the IVF envelope which are highly conserved. There is therefore practically no risk of the appearance of resistant viral strains, because these highly conserved zones correspond to peptide sequences which are very useful, even essential to the virus, and of which certain key amino acids cannot in any case be mutated without compromising the entry of the virus, and therefore its replication, in target cells.

On peut mettre en évidence l'effet d'inhibition de l'entrée du virus FIV dans ses cellules cibles par divers tests connus, en particulier le test d'inhibition de la formation de syncitia. On peut opérer par exemple de la façon décrite par S. Lombardi et al., Virology, 220,274-284 (1996), qui utilise une lignée cellulaire fibroblastoïde de rein de félin Crandell désignée en abrégé par CrFK. Les cellules sont cultivées de façon connue. On ajoute le peptide étudié, à une concentration finale variant de 0,01 à 25 u. g/ml, à des fractions de 0,8 ml contenant 2. 104 cellules CrFK, puis on ajoute le virus  The inhibition effect of the entry of the IVF virus into its target cells can be demonstrated by various known tests, in particular the test for inhibition of syncitia formation. One can operate, for example, as described by S. Lombardi et al., Virology, 220, 274-284 (1996), which uses a Feline kidney fibroblastoid cell line Crandell abbreviated as CrFK. The cells are cultured in a known manner. The studied peptide is added, at a final concentration varying from 0.01 to 25 u. g / ml, to 0.8 ml fractions containing 2.104 CrFK cells, then the virus is added

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FIV, isolé chez un chat séro-négatif à FELV (détection à l'aide du snap-test IDEXX), et multiplié de façon connue. Le sigle FELV désigne le virus de la leucémie féline. Après six jours de culture, on compte au microscope le nombre de syncitia formés. Pour les essais témoins, on ajoute, à la place du peptide testé, soit un peptide de même composition en acides aminés, mais de séquence aléatoire, soit un neuropeptide commercialisé par la firme NEOSYSTEM (Strasbourg, France) sous la dénomination SC371.  IVF, isolated from a cat negative for FELV (detection using the IDEXX snap-test), and multiplied in a known manner. FELV stands for feline leukemia virus. After six days of culture, the number of syncitia formed is counted under a microscope. For the control tests, either a peptide of the same amino acid composition but of random sequence, or a neuropeptide marketed by the company NEOSYSTEM (Strasbourg, France) under the name SC371, is added in place of the tested peptide.

Les peptides de l'invention inhibent la formation des syncitia, cette inhibition étant fonction de la dose de peptide.  The peptides of the invention inhibit the formation of syncitia, this inhibition being dependent on the dose of peptide.

Les peptides constituant le principe actif des médicaments selon l'invention correspondent à des séquences incluses dans la séquence de la protéine gp36 de FIV. Ces peptides ont une séquence peptidique contenant au moins 25 motifs peptidiques consécutifs (résidus d'acides aminés), et en particulier au moins 30 motifs peptidiques consécutifs inclus dans l'une des séquences 649-696 et 729-782 de la protéine gp36 d'une souche de FIV ou d'une séquence consensus correspondante, et lesdits peptides contiennent au plus 40 motifs peptidiques.  The peptides constituting the active principle of the medicaments according to the invention correspond to sequences included in the sequence of the protein gp36 of IVF. These peptides have a peptide sequence containing at least 25 consecutive peptide motifs (amino acid residues), and in particular at least 30 consecutive peptide motifs included in one of the sequences 649-696 and 729-782 of the protein gp36 d ' an IVF strain or a corresponding consensus sequence, and said peptides contain at most 40 peptide motifs.

Dans le Tableau 1 annexé, on a représenté les séquences 649-782 de la protéine gp36 de diverses souches connues du FIV. Ces séquences sont accessibles notamment auprès de la base de données SWISSPROT. Les codes SWISSPROT de ces souches sont mentionnés dans le Tableau 1. On peut donc aisément identifier à l'aide du Tableau 1 les séquences qui sont mentionnées dans la description de la présente demande.  In the appended Table 1, the sequences 649-782 of the gp36 protein of various known strains of IVF are shown. These sequences are accessible in particular from the SWISSPROT database. The SWISSPROT codes of these strains are mentioned in Table 1. It is therefore easy to identify, using Table 1, the sequences which are mentioned in the description of the present application.

Dans le Tableau 2 annexé, on a représenté une séquence consensus 649-782 correspondante. In the attached Table 2, a corresponding consensus sequence 649-782 is shown.

Dans les séquences qui viennent d'être mentionnées, on peut remplacer K par R et inversement, et on peut remplacer 1 par L et inversement.  In the sequences which have just been mentioned, one can replace K by R and vice versa, and one can replace 1 by L and vice versa.

Les peptides tels que définis ci-dessus peuvent être utilisés comme médicaments dans le traitement des infections à FIV chez le chat. Ils peuvent être administrés par exemple à des doses allant de 0,1 à 2 mg/kg de poids corporel.  The peptides as defined above can be used as drugs in the treatment of IVF infections in cats. They can be administered for example in doses ranging from 0.1 to 2 mg / kg of body weight.

Le médicament de l'invention, qui contient au moins un peptide tel que défini ci-dessus, et éventuellement un excipient et/ou un véhicule pharmaceutique approprié, peut être administré notamment par injection intramusculaire ou intraveineuse. A cet effet, il peut se présenter en particulier sous la forme de solution ou suspension injectable  The medicament of the invention, which contains at least one peptide as defined above, and optionally an excipient and / or a suitable pharmaceutical vehicle, can be administered in particular by intramuscular or intravenous injection. For this purpose, it can be in particular in the form of solution or suspension for injection.

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ou sous la forme de poudre lyophilisée contenant le peptide utilisé comme principe actif, avec éventuellement un adjuvant de lyophilisation classique. La poudre lyophilisée permet de reconstituer une solution ou suspension injectable.  or in the form of lyophilized powder containing the peptide used as active principle, with optionally a conventional lyophilization adjuvant. The lyophilized powder makes it possible to reconstitute an injectable solution or suspension.

L'invention concerne en outre l'utilisation d'au moins un peptide tel que défini ci-dessus dans la préparation d'un médicament destiné à lutter contre l'infection par le virus de l'immunodéficience féline chez le chat.  The invention further relates to the use of at least one peptide as defined above in the preparation of a medicament intended for combating infection with the feline immunodeficiency virus in cats.

L'exemple suivant illustre l'invention.  The following example illustrates the invention.

Exemple :
On a préparé par synthèse le peptide correspondant à la séquence 742-777 du Tableau 2. Example:
The peptide corresponding to the sequence 742-777 in Table 2 was prepared by synthesis.

On prépare une poudre lyophilisée à partir d'une solution aqueuse, stérile et apyrogène de ce peptide.  A lyophilized powder is prepared from an aqueous, sterile and pyrogen-free solution of this peptide.

La solution injectable peut être reconstituée à partir de la poudre lyophilisée avec du sérum apyrogène pour former une solution isotonique injectable.  The solution for injection can be reconstituted from the lyophilized powder with pyrogen-free serum to form an isotonic solution for injection.

On peut remplacer le peptide correspondant à la séquence 742-777 du Tableau 2 par un peptide correspondant à l'une des séquences 742-777 du Tableau 1, ou par un peptide correspondant à l'une des séquences 654-691 du Tableau 1 ou du Tableau 2, ou par un peptide correspondant à l'une des séquences 734-762 du Tableau 1 ou du Tableau 2.  The peptide corresponding to the sequence 742-777 in Table 2 can be replaced by a peptide corresponding to one of the sequences 742-777 in Table 1, or by a peptide corresponding to one of the sequences 654-691 in Table 1 or in Table 2, or with a peptide corresponding to one of the sequences 734-762 in Table 1 or in Table 2.

On peut aussi associer un peptide choisi parmi ceux des séquences 742-777 avec un peptide choisi parmi ceux des séquences 734-762 et/ou avec un peptide choisi parmi ceux des séquences 654-691 des Tableaux 1 et 2 ci-après.  It is also possible to associate a peptide chosen from those of sequences 742-777 with a peptide chosen from those of sequences 734-762 and / or with a peptide chosen from those of sequences 654-691 of Tables 1 and 2 below.

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Figure img00040001
Figure img00040001

TABLEAU 1 GP36 FIVPE HQEAIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36-PlVUl QQEAIEKVTEALKITNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP366FIVWO HQEAIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36FIVU2 HQETIEKITEALKVNNLRLVTLEHQVLVIGLKVEMEKFLYTAFKMQELGCN GP36-FIVU8 HQETIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36 FIVSD HQEALDKITFALKINNLRLVTLEHQNlVIGLKVFAIEKELYTAFAMQELGCN GP36 : FIVT2 HQQALEKITEALKINNLRLITLEHQVLVIGLRVEAIEKFLYTAFAMQELGCN GP36 FIVPE QNQFFCKIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIMDIEQNNVQG GP36-FIVUl QNQFFCKVPPELWRRYNMTINQTIWNHGNITLGEWYNQTKDLQKKFYGIIMDIEQNNVQG GP36 FIVWO QNQFFCKVPSALWERYNMTINQTIW"NHGNITLGEWYNQTXDLQQRFYEIIMDIEQNNVQG GP3 6FIVU2 QNQFFCKVPPELWQRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIMDMEQNNVQG GP36 FIVU8 QNQFFCKVPPELWKRYNMTINQTIWNHGNITLGEWYNQTKELQQKFYEIIMNIEQNNVQV GP36 FIVSD QNQFFCEIPKELWLRYNMTLNQTIWNHGNITLGEWYNQTKYLQQKFYEIIMDIEQNNVQG GP366FIVT2 QNQFFCKIPPSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEIIMDIEQNNVQG G236 FIVPE KTGIQQLQKWEDWVRWIGNIPQ GP36-FIVUl KKGLQQLQKWEDWVGWIGNIPQ GP36-FIvwa KKGLQQLQEWEDWVGWIGNIPQ GP36FIVU2 RKGLQQLQEWEDWVGWLGNIPR GP36-FIVU8 KKGLQQLQEWEDWVGWIGNIPQ GP36 FIVSD KQGLQKLQNWQDWMGWIGKIPQ GP36 : FIVT2 KTGIQQLQKWEGWIGKIPQ TABLE 1 GP36 FIVPE HQEAIEKVTGALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36-PlVUl QQEAIEKVTEALKITNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP366FIVWO HQEAIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36FIVU2 HQETIEKITEALKVNNLRLVTLEHQVLVIGLKVEMEKFLYTAFKMQELGCN GP36-FIVU8 HQETIEKVTEALKINNLRLVTLEHQVLVIGLKVEAMEKFLYTAFAMQELGCN GP36 FIVSD HQEALDKITFALKINNLRLVTLEHQNlVIGLKVFAIEKELYTAFAMQELGCN GP36: FIVT2 HQQALEKITEALKINNLRLITLEHQVLVIGLRVEAIEKFLYTAFAMQELGCN GP36 FIVPE QNQFFCKIPLELWTRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIMDIEQNNVQG GP36-FIVUl QNQFFCKVPPELWRRYNMTINQTIWNHGNITLGEWYNQTKDLQKKFYGIIMDIEQNNVQG GP36 FIVWO QNQFFCKVPSALWERYNMTINQTIW "NHGNITLGEWYNQTXDLQQRFYEIIMDIEQNNVQG GP3 6FIVU2 QNQFFCKVPPELWQRYNMTINQTIWNHGNITLGEWYNQTKDLQQKFYEIIMDMEQNNVQG GP36 FIVU8 QNQFFCKVPPELWKRYNMTINQTIWNHGNITLGEWYNQTKELQQKFYEIIMNIEQNNVQV GP36 FIVSD QNQFFCEIPKELWLRYNMTLNQTIWNHGNITLGEWYNQTKYLQQKFYEIIMDIEQNNVQG GP366FIVT2 QNQFFCKIPPSLWSMYNMTLNQTIWNHGNISLGNWYNQTRDLQNKFYEIIMDIEQNNVQG G236 FIVPE KTGIQQLQKWEDWVRWIGNIPQ GP3 6-FIVUl KKGLQQLQKWEDWVGWIGNIPQ GP36-FIvwa KKGLQQLQEWEDWVGWIGNIPQ GP36FIVU2 RKGLQQLQEWEDWVGWLGNIPR GP36-FIVU8 KKGLQQLQEWEDWVGWQWKWQWWWKQQWWKQQWWWQQ

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TABLEAU 2 HQEAIEKVTEALKINNL Numérotation 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 RLVTLEHQVLV 1 GLKVE Numérotation 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682
A M E K F L Y T A F A M Q E L G C Numérotation 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699
N Q N Q F F C K V P P E L W T R Y Numérotation 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 NMTINQTIWNHGNITLG Numérotation 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733
E W Y N Q T K D L Q Q K F Y E 1 1 Numérotation 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 M DIE Q N N V Q G K K G 1 Q Q L Numérotation 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767
Q K W E D W V G W 1 G N 1 P Q Numérotation 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782TABLE 2 HQEAIEKVTEALKINNL Numbering 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 RLVTLEHQVLV 1 GLKVE Numbering 666 667 668 669 670 671 672 673 674 675 676 677 671 672 680
AMEKFLYTAFAMQELGC Numbering 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699
NQNQFFCKVPPELWTRY Numbering 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 NMTINQTIWNHGNITLG Numbering 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 732
EWYNQTKDLQQKFYE 1 1 Numbering 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 M DIE QNNVQGKKG 1 QQL Numbering 751 752 753 754 755 756 757 758 759 760 761 762 763 764 767 764
QKWEDWVGW 1 GN 1 PQ Numbering 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782

Claims (6)

REVENDICATIONS 1. Peptides contenant au moins 25 motifs peptidiques consécutifs inclus dans l'une des séquences 649-696 et 729-782 de la protéine gp36 d'une souche de FIV, ou d'une séquence consensus correspondante, et contenant au plus 40 motifs peptidiques consécutifs de la protéine gp36 de FIV, étant entendu que dans la séquence desdits peptides, K peut être remplacé par R et inversement, et 1 peut être remplacé par L et inversement. 1. Peptides containing at least 25 consecutive peptide motifs included in one of the sequences 649-696 and 729-782 of the gp36 protein of an IVF strain, or of a corresponding consensus sequence, and containing at most 40 peptide motifs consecutive of the IVF protein gp36, it being understood that in the sequence of said peptides, K can be replaced by R and vice versa, and 1 can be replaced by L and vice versa. 2. Peptides selon la revendication 1, contenant au moins 30 motifs peptidiques consécutifs inclus dans lesdites séquences 649-696 et 729-782.  2. Peptides according to claim 1, containing at least 30 consecutive peptide motifs included in said sequences 649-696 and 729-782. 3. Peptides selon l'une quelconque des revendications précédentes, dans lesquels lesdites séquences 649-696 et 729-782 sont choisies parmi les séquences SEQ ID NO 1 à 15.  3. Peptides according to any one of the preceding claims, in which said sequences 649-696 and 729-782 are chosen from sequences SEQ ID NO 1 to 15. 4. Peptides selon l'une quelconque des revendications précédentes, contenant au moins 25 motifs peptidiques consécutifs choisis dans l'une des séquences 654-691,742- 777 et 734-762.  4. Peptides according to any one of the preceding claims, containing at least 25 consecutive peptide motifs chosen from one of the sequences 654-691,742-777 and 734-762. 5. Médicament contenant au moins un peptide tel que défini dans l'une quelconque des revendications précédentes, et éventuellement un excipient et/ou un véhicule pharmaceutique.  5. Medicament containing at least one peptide as defined in any one of the preceding claims, and optionally an excipient and / or a pharmaceutical vehicle. 6. Utilisation d'un peptide tel que défini dans l'une quelconque des revendications 1 à 4 dans la préparation d'un médicament destiné à lutter contre l'infection par le virus de l'immunodéficience féline chez le chat. 6. Use of a peptide as defined in any one of claims 1 to 4 in the preparation of a medicament intended to fight against infection by the feline immunodeficiency virus in cats.
FR0110910A 2001-08-17 2001-08-17 NOVEL PEPTIDES AND THEIR USE AS MEDICAMENTS AGAINST IVF INFECTION IN CATS Expired - Fee Related FR2828686B1 (en)

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FR0110910A FR2828686B1 (en) 2001-08-17 2001-08-17 NOVEL PEPTIDES AND THEIR USE AS MEDICAMENTS AGAINST IVF INFECTION IN CATS
FR0115424A FR2828687B1 (en) 2001-08-17 2001-11-29 NOVEL PEPTIDES AND THEIR USE AS MEDICAMENTS AGAINST IVF INFECTION IN CATS
PCT/US2002/026247 WO2003015814A1 (en) 2001-08-17 2002-08-19 New peptides and use thereof in therapeutic agents against feline fiv infection

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