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WO1997046585A2 - Fragments de leptine (proteine ob) - Google Patents

Fragments de leptine (proteine ob) Download PDF

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Publication number
WO1997046585A2
WO1997046585A2 PCT/EP1997/002968 EP9702968W WO9746585A2 WO 1997046585 A2 WO1997046585 A2 WO 1997046585A2 EP 9702968 W EP9702968 W EP 9702968W WO 9746585 A2 WO9746585 A2 WO 9746585A2
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WO
WIPO (PCT)
Prior art keywords
peptide
host cell
dna
peptides
functional derivative
Prior art date
Application number
PCT/EP1997/002968
Other languages
English (en)
Other versions
WO1997046585A3 (fr
Inventor
Kamal A. Al-Barazanji
Jonathan Robert Sanders Arch
Patrick Camilleri
William Arthur Neville
Original Assignee
Smithkline Beecham P.L.C.
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
Priority claimed from GBGB9611775.9A external-priority patent/GB9611775D0/en
Priority claimed from GBGB9618540.0A external-priority patent/GB9618540D0/en
Priority claimed from GBGB9703493.8A external-priority patent/GB9703493D0/en
Priority to EP97929156A priority Critical patent/EP0912609A2/fr
Priority to JP10500240A priority patent/JP2000512137A/ja
Priority to HU0001774A priority patent/HUP0001774A3/hu
Application filed by Smithkline Beecham P.L.C. filed Critical Smithkline Beecham P.L.C.
Priority to IL12715397A priority patent/IL127153A0/xx
Priority to NZ332879A priority patent/NZ332879A/xx
Priority to CA002257240A priority patent/CA2257240A1/fr
Priority to BR9709529A priority patent/BR9709529A/pt
Priority to AU33372/97A priority patent/AU3337297A/en
Publication of WO1997046585A2 publication Critical patent/WO1997046585A2/fr
Publication of WO1997046585A3 publication Critical patent/WO1997046585A3/fr
Priority to NO985683A priority patent/NO985683D0/no

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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
    • C07K14/575—Hormones
    • 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
    • C07K14/575—Hormones
    • C07K14/5759—Products of obesity genes, e.g. leptin, obese (OB), tub, fat
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • 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 invention relates to novel compounds, in particular to novel peptides, to compositions containing such compounds and to the use of such compounds in medicine.
  • the present invention provides a peptide or a functional derivative, analogue or variant thereof, which modulates body weight, substantially be means of modulating energy utilisation.
  • the modulation of body weight is a reduction of body weight.
  • the modulation of energy utilisation is via an enhancement of energy utilization.
  • the peptide is a fragment of an ob protein, especially human ob protein, or a functional derivative, analogue or variant thereof.
  • TIVTR ob37 -41
  • ob42 -54 INDISHTQSVSSK
  • ob55 -56 QK
  • ob57 -74 VTGLDFIPGLHPILTLSK
  • ob93 -105 NVIQISNDLENLR
  • ⁇ bl06 -115 DLLHVLAFSK
  • obi 16 -149 SCHLPWASGLETLDSLGGVLEASGYSTEVVALSR
  • obi 50- 167 LQGSLQDMLWQLDLSPGC especially ob57 -74 (VTGLDFIPGLHPILTLSK)
  • the invention includes a peptide formed from any one or more of the aforementioned particular peptides.
  • the invention includes a peptide formed from any of two contiguous members of the aforementioned particular peptides.
  • the invention also extends to the functional derivatives, analogues and variants of the peptides mentioned herein:
  • Functional derivatives includes salts and solvates of the peptides mentioned herein and also the peptides of the invention chemically modified by the attachment of groups or moieties so as to improve the physical properties, such as stability, or the therapeutic properties, for example the pharmacokinetic properties, of the protein.
  • Functional analogues includes functionally analogous peptides wherein one or more amino acids of the peptides mentioned herein are replaced with alternative amino acids.
  • Alternative amino acids includes amino acids of alternative stereochemistry to the amino acids in ob protein.
  • Functional analogues also include small molecule agonists or antagonists of the peptides mentioned herein. Such compounds may be prepared and tested according to known procedures, for example those disclosed in GB2292382.
  • Salts include pharmaceutically acceptable salts, especially pharmaceutically acceptable acid addition salts.
  • Acid addition salts of the peptides are prepared in a standard manner in a suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, hydrobromic, sulphuric, phosphoric, acetic, maleic, succinic, or methanesulphonic.
  • the acetate salt form is especially useful.
  • Certain of the compounds form inner salts or zwitterions which may be acceptable.
  • Cationic salts are prepared by treating the parent compound with an excess of an alkaline reagent, such as a hydroxide, carbonate or alkoxide containing the appropriate cation.
  • Cations such as Na + , K + , Ca ⁇ "1" and NH_ ⁇ + are examples of cations present in pharmaceutically acceptable salts.
  • Solvates include pharmaceutically acceptable solvates, such as hydrates. It will be appreciated that the invention includes both peptide and non-peptide compounds. In addition the invention includes sub-fragments of the particular peptides ob21-26, ob27-32, ob33 -36, ob37-41 , ob42 -54 , ob.55-56, ob57 -74, ob93 -105 , oblO ⁇ -115 , obi 16 -149 and obi 50 -167 , especially ob57 -74 ; or a peptide formed from any one or more, especially of any two contiguous members, of the said particular peptides; or a functional derivative, analogue or variant thereof. Suitable peptides or sub-fragments comprise at least 4 amino acids.
  • the peptides of the invention are suitably prepared by using conventional digestion methods, synthetic techniques or by use of standard expression methodology.
  • the present invention provides a process for the preparation of a peptide, or a functional derivative thereof, the process comprising the steps of: hydrolysing the peptide, especially an ob protein and in particular a human ob protein, into at least two peptide fragments; separating the peptide fragments; and optionally thereafter preparing a functional derivative thereof.
  • the hydrolysis of the protein is suitably effected by enzymic digestion, using for example trypsin.
  • the separation of the required peptide is conveniently accomplished by use of an appropriate chromatographic means, such as column chromatography.
  • reaction conditions for the treatment of the ob protein are determined by the nature of the particular reagent used, for examples when trypsin is the reagent then the reaction is normally carried out within a temperature range of 25-40°C and a pH range of 7-9, preferably at 37°C and pH 7.4.
  • the present invention provides a synthetic peptide or a functional derivative, analogue or variant thereof, which modulates body weight, substantially be means of modulating energy utilisation.
  • the peptides are prepared by the solid phase technique of Merrifield (J. ⁇ m. C em- S ⁇ £., £5_:2149 (1964)).
  • a combination of solid phase and solution synthesis may be used, as in a convergent synthesis in which di-, tri, tetra-, or penta-peptide fragments may be prepared by solid phase synthesis and either coupled or further modified by solution synthesis.
  • the side chain functional groups e.g., -NH2, -COOH, -OH,
  • ⁇ -amino groups are protected during the coupling reactions.
  • the ⁇ -amino group is temporarily protected as fluorenylmethoxycarbonyl (Fmoc) but other acid- or base- labile protecting groups can be used, e.g., t-Butoxycarbonyl (Boc).
  • the amino side chain group of ly sine is protected as t-butoxy carbonyl, benzyloxycarbonyl or p-chlorobenzyloxycarbonyl (Z or Cl-Z).
  • Acetamidomethyl, trityl, t-butyl, S-t-butyl or para-methylbenzyl (p-MBz) protection is used for cysteines.
  • Hydroxy groups are protected as butyl or benzyl ethers and carboxyl groups are protected as butyl, benzyl (Bz) or cyclohexyl esters.
  • the peptides can be synthesized either from the C-terminus or the N-terminus, preferably the former. Prior to coupling the alpha-carboxyl group (of a suitable protected amino acid) is activated. One skilled in the art can activate the protected group in a number of ways.
  • N,N' dicyclohexylcarbodiimide DCC
  • 2(lH-benzotriazol- l-yl)-l,l,3,3-tetramethyl uronium hexafluorophosphate HBTU
  • pNp p-nitrophenyl esters
  • HOBt hydroxybenzotriazole ester
  • OSu N-hydroxy succinimidyl ester
  • Solution synthesis of peptides is accomplished using conventional methods to form amide bonds.
  • a protected Boc-amino acid which has a free carboxyl group is coupled to a protected amino acid which has a free amino group using a suitable carbodiimide coupling agent, such as N,N' dicyclohexyl carbodiimide (DCC), optionally in the presence of 1 -hydroxybenzotriazole (HOBT) and dimethylamino pyridine (DMAP).
  • DCC N,N' dicyclohexyl carbodiimide
  • HOBT 1 -hydroxybenzotriazole
  • DMAP dimethylamino pyridine
  • the coupling reactions are preferably carried out at low temperature (e.g., -20°C) in such solvents as dichloromethane (DCM), dimethyl formamide (DMF), N-methyl pyrrolidone (NMP), tetrahydrofuran (THF) acetonitrile (ACN) or dioxane.
  • DCM dichloromethane
  • DMF dimethyl formamide
  • NMP N-methyl pyrrolidone
  • THF tetrahydrofuran
  • ACN acetonitrile
  • the first amino acid residue is normally attached to an insoluble polymer.
  • an insoluble polymer For example, two commonly used polymers are polystyrene (1% cross-linked with divinyl benzene) and 1% cross-linked polyacrylamide. These polymers are functionalised to contain a reactive group, e.g., -OH, -NH2 and -CH2CI to link the first amino acid of the targeted peptide (i.e., carboxy terminus). The choice of the linkage between the first amino acid and the polymer is dictated by the carboxy terminus of the peptide.
  • the synthetic peptides may be cyclized coupled using methods well known in the art.
  • an intramolecular disulphide is to be formed then the corresponding linear peptide can be completely deprotected and produced as a dimercaptan.
  • Any oxidizing agent known in the art to be capable of converting a dimercaptan to a disulphide may then be used.
  • Examplary of such agents are an alkali metal ferricyanide, (e.g., potassium or sodium ferricyanide), oxygen gas, diiodomethane or iodine.
  • the reaction is conducted in a suitable inert solvent, such as aqueous methanol or water, at temperatures from about 0 to 40°C, under high dilution.
  • the pH is usually maintained at about 7 to 8. Cyclisation may be performed upon the peptide while it is still attached to the support resin or while other functional groups are still protected, but it is preferably performed on the deprotected free peptide.
  • cysteine thiol protecting groups can be employed eg Acm and trityl.
  • Each peptide would contain one of each type arranged so that one pair of cysteines to be coupled are protected with trityl groups and the other pair with Acm. Independent removal of the trityl group from each peptide would give two separate monothiol derivatives which can be coupled by activating the thiol on one peptide with 2,2'dipyridyldisulphide and then adding the other monothiol peptide to give the bis(S- acetamido- methyl)disulphide-linked peptide.
  • the second disulphide can be obtained by direct iodine oxidation of this product as described by Kamber (B. Kamber, Helv. Chim. Acta 5.4, 927, (1971)), and Kamber et. al. (B. Kamber et. al., Helv. Chim. Acta 61, 899, (1980)).
  • Peptide chains can also be coupled using a linking group such as -
  • Coupling to the growing peptide chain is through the a carboxyl of the glutamic acid residue and removal of the Fmoc grouping on the lysine a amino group provides a starting point for addition of further amino acids.
  • N a -trityl protecting group may be employed on the glutamic acid residue and after coupling this may be removed with 80% acetic acid and N-acetylated with acetic anhydride. Further couplings may proceed as previously described.
  • N-terminal N-acetyl groups may be introduced by acetylation of the free amino proteinated by removal of the amino protecting group, with acetic anhydride.
  • C-terminal carboxamide groups are obtained by using an appropriate solid phase synthesis resin such as the Rink amide resin.
  • peptides of the invention may also be prepared using recombinant DNA techniques by expression of DNA encoding the polypeptide sequence.
  • the invention extends to a recombinant peptide or a functional derivative, analogue or variant thereof, which modulates body weight, substantially be means of modulating energy utilisation.
  • a recombinant peptide or a functional derivative, analogue or variant thereof which modulates body weight, substantially be means of modulating energy utilisation.
  • Any of the peptides mentioned herein form part of the invention as recombinant peptides.
  • the invention provides a process for preparing a compound according to the invention which process comprises expressing DNA encoding said compound in a recombinant host cell and recovering the product.
  • the DNA polymer comprising a nucleotide sequence that encodes the compound also forms part of the invention.
  • the process of the invention may be performed by conventional recombinant techniques such as described in Maniatis et. aj., Molecular Cloning - A Laboratory Manual; Cold Spring Harbor, 1982 and DNA Cloning vols I, II and III (D.M. Glover ed., IRL Press Ltd).
  • the process may comprise the steps of: i) preparing a replicable expression vector capable, in a host cell, of expressing a DNA polymer comprising a nucleotide sequence that encodes said compound; ii) transforming a host cell with said vector; iii) culturing said transformed host cell under conditions permitting expression of said DNA polymer to produce said compound; and iv) recovering said compound.
  • the invention also provides a process for preparing the DNA polymer by the condensation of appropriate mono-, di- or oligomeric nucleotide units.
  • the preparation may be carried out chemically, enzymatically, or by a combination of the two methods, in vitro or in vivo as appropriate.
  • the DNA polymer may be prepared by the enzymatic of appropriate DNA fragments, by conventional methods such as those described by D. M. Roberts ej aj in Biochemistry
  • the DNA fragments may be obtained by digestion of DNA containing the required sequences of nucleotides with appropriate restriction enzymes, by chemical synthesis, by enzymatic polymerisation on DNA or RNA templates, or by a combination of these methods. Preferably total synthesis of DNA fragments would be employed.
  • Digestion with restriction enzymes may be performed in an appropriate buffer at a temperature of 20°-70°C, preferably in a volume of 50ml or less with
  • Enzymatic polymerisation of DNA may be carried out in vitro using a DNA polymerase such as DNA polymerase I (Klenow fragment) in an appropriate buffer containing the nucleoside triphosphates dATP, dCTP, dGTP and dTTP as required at a temperature of 10°-37°C, proteinrally in a volume of 50ml or less.
  • a DNA polymerase such as DNA polymerase I (Klenow fragment) in an appropriate buffer containing the nucleoside triphosphates dATP, dCTP, dGTP and dTTP as required at a temperature of 10°-37°C, proteinrally in a volume of 50ml or less.
  • Enzymatic ligation of DNA fragments may be carried out using a DNA ligase such as T4 DNA ligase in an appropriate buffer at a temperature of 4°C to ambient, in a volume of 50ml or less.
  • a DNA ligase such as T4 DNA ligase in an appropriate buffer at a temperature of 4°C to ambient, in a volume of 50ml or less.
  • the chemical synthesis of the DNA polymer or fragments may be carried out by conventional phosphotriester, phosphite or phosphoramidite chemistry, using solid phase techniques such as those described in 'Chemical and Enzymatic Synthesis of Protein Fragments - A Laboratory Manual' (ed. H.G. Gassen and A. Lang), Verlag
  • the DNA polymer is preferably prepared by ligating two or more DNA molecules which together comprise a DNA sequence encoding the compound.
  • the DNA molecules may be obtained by the digestion with suitable restriction enzymes of vectors carrying the required coding sequences.
  • the precise structure of the DNA molecules and the way in which they are obtained depends upon the structure of the desired product.
  • the design of a suitable strategy for the construction of the DNA molecule coding for the compound is a routine matter for the skilled worker in the art.
  • the expression of the DNA polymer encoding the compound in a recombinant host cell may be carried out by means of a replicable expression vector capable, in the host cell, of expressing the DNA polymer.
  • the expression vector is novel and also forms part of the invention.
  • the replicable expression vector may be prepared in accordance with the invention, by cleaving a vector compatible with the host cell to provide a linear DNA segment having an intact replicon, and combining said linear segment with one or more DNA molecules which, together with said linear segment, encode the compound, under ligating conditions.
  • the ligation of the linear segment and more than one DNA molecule may be carried out simultaneously or sequentially as desired.
  • the DNA polymer may be preformed or formed during the construction of the vector, as desired.
  • the preparation of the replicable expression vector may be carried out conventionally with appropriate enzymes for restriction, polymerisation and ligation of the DNA, by procedures described in, for example, Maniatis ⁇ l al., cited above. Polymerisation and ligation may be performed as described above for the preparation of the DNA polymer. Digestion with restriction enzymes may be performed in an appropriate buffer at a temperature of 20°-70°C, proteinrally in a volume of 50ml or less with O.l-lOmg DNA.
  • the recombinant host cell is prepared, in accordance with the invention, by transforming a host cell with a replicable expression vector of the invention under transforming conditions.
  • Suitable transforming conditions are conventional and are described in, for example, Maniatis ⁇ l ai., cited above, or "DNA Cloning" Vol. II, D.M. Glover ed., IRL Press Ltd, 1985.
  • the invention also extends to a vector comprising a compound of the invention.
  • the invention also extends to a host cell transformed with a replicable expression vector of the invention.
  • the DNA polymer may be assembled into vectors designed for isolation of stable transformed mammalian cell lines expressing the product; e.g. bovine papillomavirus vectors or amplified vectors in Chinese hamster ovary cells (DNA cloning Vol.II D.M. Glover ed. IRL Press 1985; Kaufman, R.J. ⁇ l al., Molecular and Cellular Biology 5, 1750-1759, 1985; Pavlakis G.N. and Hamer, D.H., Proceedings of the National Academy of Sciences (USA) 80, 397-401, 1983; Goeddel, D.V. si al, European Patent Application No. 0093619, 1983).
  • bovine papillomavirus vectors or amplified vectors in Chinese hamster ovary cells
  • the compounds of the invention are considered to be capable of modulating body weight substantially by means of enhancing energy utilization and are therefore of potential use in the treatment of nutritional and metabolic disorders, particularly obesity and diabetes.
  • the invention also provides a method for the treatment of nutritional and metabolic disorders, which method comprises the administration of an effective, pharmaceutically acceptable and non-toxic amount of a compound of the invention.
  • the invention therefore further provides a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier.
  • the active compound will normally be employed in the form of a pharmaceutical composition in association with a human or veterinary pharmaceutical carrier, diluent and/or excipient, although the exact form of the composition will depend on the mode of administration.
  • the active compound may, for example, be employed in the form of tablets, capsules, lozenges or syrups for oral administration; in the form of snuff, aerosol or nebulisable solution for inhalation; in the form of sterile solutions for parenteral administration, or in the form of creams, lotions, liniments, gels, ointments or sprays for topical administration.
  • Parenteral routes of administration include intravenous, intramuscular, subcutaneous, transcutaneous and intraperitoneal administration.
  • Solid compositions suitable for oral administration may be obtained by conventional methods of blending, filling, granulation, tabletting or the like.
  • Repeated blending operations may be used to distribute the active agent throughout those compositions employing large quantities of fillers.
  • composition may be implanted subcutaneously, for example in the form of a compressed tablet or slow release capsule.
  • Fluid unit dosage forms are prepared utilising the compound and a pyrogen-free sterile vehicle.
  • the compound depending on the vehicle and concentration used, can be either dissolved or suspended in the vehicle. Solutions may be used for all forms of parenteral administration, and are particularly used for intravenous infection. In preparing solutions the compound can be dissolved in the vehicle, the solution being made isotonic if necessary by addition of sodium chloride and sterilised by filtration through a sterile filter using aseptic techniques before filling into suitable sterile vials or ampoules and sealing. Alternatively, if solution stability is adequate, the solution in its sealed containers may be sterilised by autoclaving. Advantageously additives such as buffering, solubilising, stabilising, preservative or bactericidal, suspending or emulsifying agents and/or local anaesthetic agents may be dissolved in the vehicle.
  • the compound may be isolated in a sterile state or alternatively it may be sterilised after isolation, e.g. by gamma irradiation.
  • a suspending agent for example polyvinylpyrrolidone is included in the composition to facilitate uniform distribution of the compound.
  • Rats are pre-treated with Synulox (0.1 ml/1 OOg) approx 1 hour before anaesthesia, and then anaesthetised with Domitor (0.04ml/100g i.m.) and sublimase (0.9ml/100g i.p.)
  • Domitor 0.04ml/100g i.m.
  • sublimase 0.9ml/100g i.p.
  • Each rat has a cannula implanted stereotaxically into the lateral brain ventricle under sterile conditions. Anaesthesia is then reversed using Antisedan and Nubain (50% v/v : 50% v/v 0.02ml/l OOg) I.P. After surgery each rat receives 0.05 ml Zenecarp.
  • Angiotensin II (100ng/5 ⁇ l) was injected icv and water intake was monitored for 5 min after injection.
  • mice 24 hour food intake was measured on day 5 and 6 after surgery.
  • the animals were divided according to their body weight into 3 groups (a,b and c, 8 rats per group) and then fasted overnight.
  • rats On the day of experiment (day 7) rats were injected icv as follows:: group a-vehicle (PBS, phosphate buffer solution, 5 ⁇ l /rat); group b- human leptin (11.5 ⁇ g/5 ⁇ l); and group c- leptin tryptic digest (30 ⁇ g/5 ⁇ l).
  • group a-vehicle PBS, phosphate buffer solution, 5 ⁇ l /rat
  • group b- human leptin (11.5 ⁇ g/5 ⁇ l
  • group c- leptin tryptic digest (30 ⁇ g/5 ⁇ l).
  • mice were injected as follows: group a-vehicle (PBS 5 ⁇ l /rat); group b-human leptin (8.75 ⁇ g/5 ⁇ l); group c-murine leptin ( 10 ⁇ g/5 ⁇ l); group d-ob 57 -74 , sequence VTGLDFIPGLHPILTLSK (3.33 ⁇ g/5 ⁇ l);

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Obesity (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Toxicology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Diabetes (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • Child & Adolescent Psychology (AREA)
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  • Veterinary Medicine (AREA)
  • Endocrinology (AREA)
  • Hematology (AREA)
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  • Gastroenterology & Hepatology (AREA)
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  • Genetics & Genomics (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne une leptine, un peptide ob, ou un dérivé fonctionnel, un analogue ou un variant de ceux-ci, modulant le poids corporel essentiellement au moyen d'une modulation de l'utilisation de l'énergie. Elle concerne également une composition pharmaceutique contenant un tel composé, un procédé de préparation de celui-ci, ainsi que l'utilisation de ce composé en médecine.
PCT/EP1997/002968 1996-06-06 1997-06-04 Fragments de leptine (proteine ob) WO1997046585A2 (fr)

Priority Applications (9)

Application Number Priority Date Filing Date Title
AU33372/97A AU3337297A (en) 1996-06-06 1997-06-04 Fragments of leptin (ob protein)
BR9709529A BR9709529A (pt) 1996-06-06 1997-06-04 Fragmentos de leptina (proteína ob)
CA002257240A CA2257240A1 (fr) 1996-06-06 1997-06-04 Fragments de leptine (proteine ob)
JP10500240A JP2000512137A (ja) 1996-06-06 1997-06-04 レプチン（ｏｂタンパク質）のフラグメント
HU0001774A HUP0001774A3 (en) 1997-02-20 1997-06-04 Fragments of leptin (ob protein)
EP97929156A EP0912609A2 (fr) 1996-06-06 1997-06-04 Fragments de leptine (proteine ob)
IL12715397A IL127153A0 (en) 1996-06-06 1997-06-04 Fragments of leptin (ob protein)
NZ332879A NZ332879A (en) 1996-06-06 1997-06-04 Use of fragments of leptin (ob protein) in medicaments for treating obesity or diabetes
NO985683A NO985683D0 (no) 1996-06-06 1998-12-04 Fragmenter av leptin (ob protein)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
GBGB9611775.9A GB9611775D0 (en) 1996-06-06 1996-06-06 Novel compounds
GBGB9618540.0A GB9618540D0 (en) 1996-09-05 1996-09-05 Novel compounds
GB9618540.0 1997-02-20
GB9703493.8 1997-02-20
GBGB9703493.8A GB9703493D0 (en) 1997-02-20 1997-02-20 Novel compounds
GB9611775.9 1997-02-20

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US09194866 A-371-Of-International 1998-12-04
US09/844,774 Continuation US20020037553A1 (en) 1996-06-06 2001-04-27 Fragments of leptin (ob protein)

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WO1997046585A2 true WO1997046585A2 (fr) 1997-12-11
WO1997046585A3 WO1997046585A3 (fr) 1998-04-23

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EP (1) EP0912609A2 (fr)
JP (1) JP2000512137A (fr)
KR (1) KR20000016402A (fr)
CN (1) CN1221426A (fr)
AR (1) AR008765A1 (fr)
AU (1) AU3337297A (fr)
BR (1) BR9709529A (fr)
CA (1) CA2257240A1 (fr)
CZ (1) CZ397898A3 (fr)
IL (1) IL127153A0 (fr)
NO (1) NO985683D0 (fr)
NZ (1) NZ332879A (fr)
PL (1) PL330361A1 (fr)
TR (1) TR199802534T2 (fr)
WO (1) WO1997046585A2 (fr)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000011173A1 (fr) * 1998-08-21 2000-03-02 Albany Medical College Peptides de type leptine
WO2000021574A2 (fr) 1998-10-14 2000-04-20 Amgen Inc. Double polyethyleneglycolation de proteines dirigee sur site pour augmenter la bioactivite et la biocompatibilite
WO2001021647A3 (fr) * 1999-09-22 2002-03-07 Genset Sa Procedes de criblage de composes qui modulent l'interaction de la lsr-leptine et utilisations de ces composes dans la prevention et le traitement des maladies liees a l'obesite
US7183254B2 (en) 2001-10-22 2007-02-27 Amgen, Inc. Use of leptin for treating human lipoatrophy and method of determining predisposition to said treatment
US7208572B2 (en) 1998-08-21 2007-04-24 Albany Medical College Leptin-related peptides
GB2451858A (en) * 2007-08-15 2009-02-18 Sergiy Konovchuk Methods of reducing body fat and treating obesity
US7521258B2 (en) 1994-08-17 2009-04-21 The Rockefeller University Methods of detecting, measuring, and evaluating modulators of body weight in biological samples, and diagnostic, monitoring, and therapeutic uses thereof
ES2319048A1 (es) * 2007-06-20 2009-05-01 Universitat De Les Illes Balears Uso de leptina en el tratamiento de alteraciones en los habitos alimentarios.
US7575878B2 (en) 2004-11-18 2009-08-18 Vib Vzw Methods of inhibiting leptin-induced signaling with fibronectin III domain antibodies
US7629315B2 (en) 2005-03-09 2009-12-08 University Of Pittsburgh Of The Commonwealth System Of Higher Education Compositions for blocking the inhibitory effect of human CRP on human leptin
WO2009147211A1 (fr) * 2008-06-04 2009-12-10 Biovitrum Ab (Publ) Nouveaux composés v
WO2009147216A1 (fr) * 2008-06-04 2009-12-10 Biovitrum Ab (Publ) Nouveaux dérivés de pyridine convenant comme mimétiques des modulateurs du récepteur de la leptine
WO2010118384A2 (fr) 2009-04-10 2010-10-14 Amylin Pharmaceuticals, Inc. Composés agonistes de l'amyline pour mammifères ayant une carence en oestrogènes
US7851471B2 (en) 2007-12-05 2010-12-14 Astrazeneca Ab (Publ) Compounds I
US7855270B2 (en) 2000-05-22 2010-12-21 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Receptor-based interaction trap
EP2330124A2 (fr) 2005-08-11 2011-06-08 Amylin Pharmaceuticals Inc. Polypeptides hybrides ayant des propriétés sélectionnables
EP2330125A2 (fr) 2005-08-11 2011-06-08 Amylin Pharmaceuticals, Inc. Polypeptides hybrides ayant des propriétés sélectionnables
EP2390264A1 (fr) 2005-02-11 2011-11-30 Amylin Pharmaceuticals Inc. Analogues de GIP et polypeptides hybrides de GIP aux propriétés sélectionnables
US8093248B2 (en) 2007-12-05 2012-01-10 Astrazeneca Ab (Publ) Compounds useful for the treatment of conditions associated with weight gain
EP2417980A1 (fr) 2004-02-11 2012-02-15 Amylin Pharmaceuticals Inc. Polypeptides hybrides présentant des propriétés pouvant être choisies
US8283307B2 (en) 2003-09-08 2012-10-09 Merck Serono Sa Treatment of fibrotic disease
US20130109619A1 (en) * 2010-05-11 2013-05-02 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Peptide-based inhibitor of interleukin-10 or interferon-gamma signaling
US8889622B2 (en) 2007-07-25 2014-11-18 Washington University Methods of inhibiting seizure in a subject
US11535659B2 (en) 2010-09-28 2022-12-27 Amryt Pharmaceuticals Inc. Engineered polypeptides having enhanced duration of action

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CA2335107C (fr) 1998-07-28 2010-01-19 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Induction de genes a mediation par la leptine
CN104829707B (zh) * 2015-05-06 2017-12-19 广东省生物资源应用研究所 一条cd环和e螺旋区突变的瘦素活性肽及其编码基因和应用
CN110183530A (zh) * 2019-06-21 2019-08-30 深圳市亚辉龙生物科技股份有限公司 瘦素免疫原、杂交瘤细胞、单克隆抗体、多克隆抗体及应用

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US6309853B1 (en) * 1994-08-17 2001-10-30 The Rockfeller University Modulators of body weight, corresponding nucleic acids and proteins, and diagnostic and therapeutic uses thereof
AU4766596A (en) * 1995-01-31 1996-08-21 Eli Lilly And Company Ob gene product antibodies
EP0836620A1 (fr) * 1995-01-31 1998-04-22 Eli Lilly And Company Proteines anti-obesite
US5552523A (en) * 1995-01-31 1996-09-03 Eli Lilly And Company Anti-obesity proteins
WO1996031526A1 (fr) * 1995-04-06 1996-10-10 Amylin Pharmaceuticals, Inc. Agents antiobesite
GB9509164D0 (en) * 1995-05-05 1995-06-28 Smithkline Beecham Plc Novel compounds

Cited By (42)

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US7521258B2 (en) 1994-08-17 2009-04-21 The Rockefeller University Methods of detecting, measuring, and evaluating modulators of body weight in biological samples, and diagnostic, monitoring, and therapeutic uses thereof
US7208572B2 (en) 1998-08-21 2007-04-24 Albany Medical College Leptin-related peptides
WO2000011173A1 (fr) * 1998-08-21 2000-03-02 Albany Medical College Peptides de type leptine
US7786265B2 (en) 1998-08-21 2010-08-31 Albany Medical College Isolated antibodies against biologically active leptin-related peptides
AU772278B2 (en) * 1998-08-21 2004-04-22 Clf Medical Technology Acceleration Program, Inc. Leptin-related peptides
US6777388B1 (en) 1998-08-21 2004-08-17 Clf Medical Technology Acceleration Program, Inc. Leptin-related peptides
US7790683B2 (en) * 1998-08-21 2010-09-07 Albany Medical College Leptin-related peptides
US8067545B2 (en) 1998-08-21 2011-11-29 Albany Medical College Isolated antibodies against biologically active leptin-related peptides
US8022189B2 (en) 1998-08-21 2011-09-20 Albany Medical College Isolated antibodies against biologically active leptin-related peptides
US7186694B2 (en) 1998-08-21 2007-03-06 Albany Medical College Leptin-related peptides
WO2000021574A2 (fr) 1998-10-14 2000-04-20 Amgen Inc. Double polyethyleneglycolation de proteines dirigee sur site pour augmenter la bioactivite et la biocompatibilite
US6420339B1 (en) 1998-10-14 2002-07-16 Amgen Inc. Site-directed dual pegylation of proteins for improved bioactivity and biocompatibility
US7470669B2 (en) 1999-09-22 2008-12-30 Serono Genetics Institute, S.A. Isolated polypeptide comprising a leptin fragment
WO2001021647A3 (fr) * 1999-09-22 2002-03-07 Genset Sa Procedes de criblage de composes qui modulent l'interaction de la lsr-leptine et utilisations de ces composes dans la prevention et le traitement des maladies liees a l'obesite
US7855270B2 (en) 2000-05-22 2010-12-21 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Receptor-based interaction trap
US8003757B2 (en) 2000-05-22 2011-08-23 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Receptor-based interaction trap
EP2219031A1 (fr) 2001-10-22 2010-08-18 Amgen, Inc. Utilisation de Leptine pour traiter la Lipoatrophine humaine et procédé pour déterminer une prédisposition à ce traitement
US8318666B2 (en) 2001-10-22 2012-11-27 Amgen, Inc. Use of leptin to treat metabolic abnormalities associated with lipoatrophy
US7183254B2 (en) 2001-10-22 2007-02-27 Amgen, Inc. Use of leptin for treating human lipoatrophy and method of determining predisposition to said treatment
US8283307B2 (en) 2003-09-08 2012-10-09 Merck Serono Sa Treatment of fibrotic disease
EP2417980A1 (fr) 2004-02-11 2012-02-15 Amylin Pharmaceuticals Inc. Polypeptides hybrides présentant des propriétés pouvant être choisies
EP2422807A2 (fr) 2004-02-11 2012-02-29 Amylin Pharmaceuticals Inc. Polypeptides hybrides présentant des propriétés pouvant être choisies
EP2422806A2 (fr) 2004-02-11 2012-02-29 Amylin Pharmaceuticals Inc. Polypeptides hybrides présentant des propriétés pouvant être choisies
US7575878B2 (en) 2004-11-18 2009-08-18 Vib Vzw Methods of inhibiting leptin-induced signaling with fibronectin III domain antibodies
EP2390264A1 (fr) 2005-02-11 2011-11-30 Amylin Pharmaceuticals Inc. Analogues de GIP et polypeptides hybrides de GIP aux propriétés sélectionnables
EP2392595A1 (fr) 2005-02-11 2011-12-07 Amylin Pharmaceuticals Inc. Analogues de GIP et polypeptides hybrides de GIP aux propriétés sélectionnables
US7629315B2 (en) 2005-03-09 2009-12-08 University Of Pittsburgh Of The Commonwealth System Of Higher Education Compositions for blocking the inhibitory effect of human CRP on human leptin
EP2330124A2 (fr) 2005-08-11 2011-06-08 Amylin Pharmaceuticals Inc. Polypeptides hybrides ayant des propriétés sélectionnables
EP2330125A2 (fr) 2005-08-11 2011-06-08 Amylin Pharmaceuticals, Inc. Polypeptides hybrides ayant des propriétés sélectionnables
ES2319048B1 (es) * 2007-06-20 2010-02-10 Universitat De Les Illes Balears Uso de leptina en el tratamiento de alteraciones en los habitos alimentarios.
ES2319048A1 (es) * 2007-06-20 2009-05-01 Universitat De Les Illes Balears Uso de leptina en el tratamiento de alteraciones en los habitos alimentarios.
US8889622B2 (en) 2007-07-25 2014-11-18 Washington University Methods of inhibiting seizure in a subject
GB2451858A (en) * 2007-08-15 2009-02-18 Sergiy Konovchuk Methods of reducing body fat and treating obesity
US7851471B2 (en) 2007-12-05 2010-12-14 Astrazeneca Ab (Publ) Compounds I
US8093248B2 (en) 2007-12-05 2012-01-10 Astrazeneca Ab (Publ) Compounds useful for the treatment of conditions associated with weight gain
WO2009147216A1 (fr) * 2008-06-04 2009-12-10 Biovitrum Ab (Publ) Nouveaux dérivés de pyridine convenant comme mimétiques des modulateurs du récepteur de la leptine
WO2009147211A1 (fr) * 2008-06-04 2009-12-10 Biovitrum Ab (Publ) Nouveaux composés v
CN102245183A (zh) * 2008-06-04 2011-11-16 阿斯利康(瑞典)有限公司 新化合物v
WO2010118384A2 (fr) 2009-04-10 2010-10-14 Amylin Pharmaceuticals, Inc. Composés agonistes de l'amyline pour mammifères ayant une carence en oestrogènes
US20130109619A1 (en) * 2010-05-11 2013-05-02 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Peptide-based inhibitor of interleukin-10 or interferon-gamma signaling
US9475839B2 (en) * 2010-05-11 2016-10-25 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Peptide-based inhibitor of interleukin-10 or interferon-gamma signaling
US11535659B2 (en) 2010-09-28 2022-12-27 Amryt Pharmaceuticals Inc. Engineered polypeptides having enhanced duration of action

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CA2257240A1 (fr) 1997-12-11
AU3337297A (en) 1998-01-05
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AR008765A1 (es) 2000-02-23
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KR20000016402A (ko) 2000-03-25
NZ332879A (en) 2000-09-29
WO1997046585A3 (fr) 1998-04-23
BR9709529A (pt) 1999-08-10
CN1221426A (zh) 1999-06-30
CZ397898A3 (cs) 1999-04-14
EP0912609A2 (fr) 1999-05-06
NO985683L (no) 1998-12-04
IL127153A0 (en) 1999-09-22
NO985683D0 (no) 1998-12-04
JP2000512137A (ja) 2000-09-19

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