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WO1995006411A1 - Procede et composition de perte de poids - Google Patents

Procede et composition de perte de poids Download PDF

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Publication number
WO1995006411A1
WO1995006411A1 PCT/US1994/009799 US9409799W WO9506411A1 WO 1995006411 A1 WO1995006411 A1 WO 1995006411A1 US 9409799 W US9409799 W US 9409799W WO 9506411 A1 WO9506411 A1 WO 9506411A1
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WO
WIPO (PCT)
Prior art keywords
cells
uncoupling protein
gene
protein
mitochondrial uncoupling
Prior art date
Application number
PCT/US1994/009799
Other languages
English (en)
Inventor
David Kagan
Original Assignee
Lipocyte, Inc.
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 Lipocyte, Inc. filed Critical Lipocyte, Inc.
Priority to AU77178/94A priority Critical patent/AU7717894A/en
Publication of WO1995006411A1 publication Critical patent/WO1995006411A1/fr

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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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • the present invention relates to a method of effecting weight reduction in warm-blooded animals, particularly in humans.
  • the invention relates to a method of effecting weight reduction in which the gene coding for mitochondrial uncoupling protein (UCP) is isolated and inserted into mammalian cells to instigate thermogenesis.
  • UCP mitochondrial uncoupling protein
  • brown adipose tissue appears to contribute to heat production by means of oxidative processes which occur in the tissue itself, and also by release of stored lipids into circulation for use by other tissues.
  • brown adipose tissue The intracellular compositions of brown adipose tissue is just what one might expect to find in a tissue that has as its specialty, rapid heat production or thermogenesis. It has been noted from chemical studies that brown fat has an unusually high content of components of an electron transfer system. Present in high concentrations are mitochondria, cytochro e c and coenzyme Q (ubiguinone) which are believed to contribute to a high rate of oxygen consumption. Of interest, is that the heat production of brown adipose tissue of the mouse and rat, is much less than that of hibernating species such as the ground squirrel. Thermogenesis in brown adipose tissue involves the participation of many enzymes and proteins in the cell.
  • the mitochondrial uncoupling protein (UCP) , also referred to as "thermogenin”, is believed to be the brown adipose tissue specific protein necessary for uncoupled heat production in the mitochondria.
  • UCP mitochondrial uncoupling protein
  • thermoogenin The mitochondrial uncoupling protein
  • the isolation of a cDNA clone for UCP as well as other cDNAs derived from brown adipose tissue RNAs has been reported by A. Jacobsson et al., in Journal of Biological Chemistry. Vol. 260, No. 30, pages 16250-16254 (1985).
  • Brown adipocytes upon stimulation by nerve terminals from signals mediated by the hypothalamus, undergo coordinate changes in the expression of several genes which include enzymes of energy and lipid metabolism as well as the mitochondrial uncoupling protein. This protein is believed to be essential for heat production by the mitochondria.
  • thermogenesis in brown adipose tissue requires the participation of many enzymes and proteins in the cell.
  • Jacobsson et al. indicated that as of 1985, only one brown adipocyte-specific gene product had been identified.
  • the present invention relates to a method for effecting weight reduction, particularly in humans, which utilizes the mitochondrial uncoupling protein.
  • the invention also relates to pharmaceutical compositions containing mitochondrial uncoupling protein-containing cells which can be administered to humans to effect weight reduction. Processes for the preparation of such compositions are also encompassed by the invention.
  • the method of effecting weight reduction comprises administering to warm-blooded animals such as humans, a weight reducing effective amount of a composition comprised of a biologically acceptable carrier and mammalian cells in which the gene for the mitochondrial uncoupling protein has been expressed.
  • a weight reducing effective amount of a composition comprised of a biologically acceptable carrier and mammalian cells in which the gene for the mitochondrial uncoupling protein has been expressed.
  • the mitochondrial uncoupling protein which as indicated is also known as "thermogenin" is a 32 kd protein which is produced by brown fat cells.
  • the protein causes the mitochondria of brown fat cells to uncouple, converting energy into heat. The consumption of calories results in a weight decrease due to the high energy consumption.
  • Brown fat is known as the flash heater, since it plays an important role in the central heating system that functions in an animal's arousal from hibernation and provides nonshivering thermogenesis in cold environments. Brown fat is responsive to adrenergic stimuli and is specialized for sympathetically regulated heat production associated with uncoupled oxidative phosphorylation in which energy provided by metabolic substrates, such as free fatty acids is used to produce heat.
  • thermogenin The nucleotide and amino acid sequence for this protein is known and several variants of thermogenin are known for different species of animals.
  • Frederic Bouillard et al. "Complete cDNA-derived Amino Acid Sequence of Rat Brown Fat Uncoupling Protein", J. Bio. Che . , Vol. 261, No. 4, pages 1487- 1490 (February 5, 1986) discloses the nucleotide sequence and amino acid sequence of rat uncoupling protein and its similarities with hamster uncoupling protein.
  • the mouse, hamster and rat proteins all have 306 amino acids.
  • the mouse protein is indicated to differ from the rat in 8 amino acids and from the hamster in 24 amino acids.
  • Fig. 1 herein provides a schematic diagram of rat gene for mitochondrial uncoupling protein.
  • Sequence Listing No. 1 of the present application shows the amino acid sequence of the human uncoupling protein gene.
  • the human and rat uncoupling protein are 79% homologous and a single probe can be used to probe for both species. Because of the functional similarity of uncoupling proteins of different species, any species may be used as a source of the uncoupling protein gene. Accordingly, the source of the uncoupling protein gene may come from human, mouse, rat, bovine, hamster and other sources. Mouse and rat uncoupling protein are preferred because brown adipose tissue is easier to obtain from these species.
  • the nucleotide sequence of mouse uncoupling protein is disclosed in L.P. Kozak et al., "The Mitochondrial Uncoupling Protein Gene", J. Biol. Che .. Vol. 263, pp. 12274-12277 (1988) .
  • the gene from any of the sources identified above may be isolated by probing a genomic library.
  • a suitable DNA probe is shown in Sequence Listing No. 2. This probe represents a highly homologous sequence between human and rat mitochondrial uncoupling protein and can be used for either species.
  • the clones isolated may be counter-screened against brown adipose tissue oligo-dT RNA prepared from cold adapted rats to verify homology. On Northern blot the RNA runs at 16s and should translate a 32 Kd protein corresponding to mitochondrial uncoupling protein.
  • the protein sequence for human mitochondrial uncoupling protein is shown in Sequence Listing No. 1.
  • the source of the brown adipose tissue is, for example, the interscapular brown fat pad in mice or peri-renal fat in humans.
  • Brown adipocytes obtained from these sources are polygonal or fibroblastic with many fat droplets within the cytoplasm. They are readily differentiated from white adipocytes which contain only one or two fat droplets per cell.
  • Brown fat precursor cells may be isolated from the location of such cells by digesting the tissue with collagenase and subsequent growth in a culture medium supplemented with a solution of new-born calf serum and insulin.
  • the tissue may be obtained from rats or mice by aseptically dissecting the interscapular fat pad. The tissue is minced and incubated in a 0.2% collagenase solution and incubated at 37"C with gentle shaking. The digested slurry is filtered through two nylon screens (250u and 25u) to remove connective tissue and undigested material. The filtered material is then washed in two changes of medium by centrifuging at 200g for 5 minutes.
  • the cells are resuspended in Dulbecco's Modified Eagles Medium (DMEM) with non-essential amino acids supplemented with 10% fetal bovine serum, suitable antibiotics (e.g. penicillin- streptomycin or gentamicin) and lOU/ml insulin and plated onto tissue culture flasks.
  • DMEM Dulbecco's Modified Eagles Medium
  • suitable antibiotics e.g. penicillin- streptomycin or gentamicin
  • lOU/ml insulin plated onto tissue culture flasks.
  • the cells have a polygonal or fibroblastic shape and contain numerous lipid vesicles.
  • tissue explant For brown adipocytes, the interscapular fat would be dissected. The sample is placed on a dry tissue culture plate and allowed to rest undisturbed for about 10 minutes. The medium described above is carefully added to the culture plate so as not to dislodge the tissue. After several days, the cells migrate out of the tissue and the tissue is then removed. Various cell types including endothelial cells, fibroblasts and smooth muscle may be isolated with this method.
  • the gene sequence which encodes the mitochondrial uncoupling protein is cloned into a suitable expression vector or plasmid of bacterial origin by known techniques and the vector or plasmid is inserted into a suitable host cell.
  • the vectors and host cells may be of mammalian, avian, reptilian, insect, yeast and bacterial origin.
  • the expression vectors which may be used as a recipient for the gene sequence are known and readily available from several sources and include those which are generally applicable to most eukaryotic cells.
  • the preferred vectors have expression which are both constitutive and stable and include pDR2 which is available from Clontech Laboratories, Inc.
  • Other vectors which may be used are pAD3, pCMV3, pSV? and pTK/?, are also available from, for example, Clontech Laboratories, Inc.
  • the vector After construction of an expression vector containing the cDNA for the mitochondrial uncoupling protein, the vector is inserted in a host cell by one of several means such as transfection, electroporation, microinjection, viral infection and the like. Promoters to be employed include RSV, Ad2, HSV, chick beta-actin, SV40, CMV and the like.
  • the host cells which are employed in the present inventions include fibroblasts, white adipocytes and the like.
  • the cells which may be of mammalian, avian or reptilian origin, containing the protein can be formulated into pharmaceutical compositions for administering to a patient desiring weight reduction, by incorporating the cells into a pharmaceutically acceptable carrier.
  • the preferred carriers are those which provide physical protection for the cells. Physical protection is desirable to prevent rupture of the cells when they are transported and injected into the patient.
  • An example of a preferred carrier is a solution of collagen of the type that has been used in recent years in collagen replacement therapy.
  • Other carriers which may be used include isotonic saline which provides osmotic protection for the cells.
  • the composition of the present invention may also include known additives such as preservatives, antioxidants and the like.
  • the presence of the mitochondrial uncoupling protein may be determined by such methods as enzyme linked immunoabsorbent assay and western blot analysis as disclosed in Ellen L.P. Chan et al., "Measurement of Brown Adipose Tissue Uncoupling Protein by Enzyme Linked Immunosorbent Assay", Bioche . Int. , Vol. 23, No. 2, pp. 291-298 (January 1991) and Mary F. Henningfield et al., "Immunochemical Detection and Quantification of Brown Adipose Tissue Uncoupling Protein", Biochem. Cell. Bio., Vol. 65, pp. 245-251 (1987) .
  • Mitochondria are isolated from transfected cells by homogenizing in 5 times the cell pellet volume of 0.25M sucrose with 5mM K-TES (pH 7.2) in a centrifuge at 8,500g for 10 minutes.
  • the pellet contains cell debris, nuclei and mitochondria and is resuspended to the original volume in the same medium.
  • the resulting mixture is centrifuged at 700g for 10 minutes to pellet the nuclei and debris which is discarded.
  • the supernatant is centrifuged at 8,500g for 10 minutes to pellet the mitochondria.
  • Mitochondrial uncoupling protein is isolated from brown adipose tissue mitochondria or from transfected cells as prepared above.
  • 500mg of mitochondrial protein is resuspended in 30ml (2% Brij , 20mM Na-T-SO,,, 10 mM MOPS, pH 6.7) by mixing well for 5 minutes at 0°C.
  • the resulting suspension is centrifuged at 800g for 5 minutes and the supernatant discarded.
  • the resulting pellet is resuspended in 20ml of 20mM Na 2 S0 4 and lOmM MOPS at a pH of 6.1 and centrifuged at 8,000g for 5 minutes.
  • the resulting pellet is shaken for 5 minutes in 30ml of 3% Triton-XlOO, 30mM Na 2 S0 4 , and 15mM of MOPS at a pH of 6.7.
  • the resulting suspension is centrifuged at 10,000g for 20 minutes.
  • the supernatant is added to a 70ml (3x10cm) hydroxylapatite column pre-equilibrated with 30mM Na 2 S0 4 and 15mM MOPS at a pH of 6.7 and eluted with the same medium.
  • the first 30ml are pooled and concentrated by pressure dialysis to a volume of from 5 to 8ml.
  • the resulting material can be used to coat microtiter plates for an enzyme linked immunosorbent assay as described hereinafter.
  • Rabbits are immunized with rat-mitochondrial uncoupling protein and blood samples containing anti-rat- mitochondrial uncoupling protein antiserum are obtained.
  • primary polyclonal rabbit anti-thermogenin antibody serum can be obtained from East Acres Biologicals of Southbridge, Massachusetts.
  • the mitochondrial uncoupling protein is subjected to sodium dodecyl sulfate-polyacryla ide gel electrophoresis to obtain a gel containing the protein.
  • the protein is transferred from the gel onto nitrocellulose paper by electroblotting.
  • the paper is then incubated with antiserum produced as described above.
  • a paper containing the same protein is incubated with normal rabbit serum as a negative control.
  • An enzyme linked immunosorbent assay can be developed by adsorbing purified mitochondrial uncoupling protein to microtiter plates. After washing to remove unbound protein and blocking with bovine serum albumin, a dilute rabbit anti-mitochondrial uncoupling protein serum is added to the appropriate wells. The unbound antibody is then washed and an anti-rabbit serum conjugated to horseradish peroxidase or other suitable reporter molecule is added. The substrate is washed and developed. High titers of mitochondrial uncoupling protein will be reflected by high substrate conversion.
  • the function of the mitochondrial uncoupling protein employed in the present invention may be determined by first isolating mitochondria from transfected cells and then isolating the mitochondrial uncoupling protein as described above.
  • the function of the mitochondrial uncoupling protein can be verified by photoaffinity labelling of the regulatory site of energy dissipation as described in G.M. Heaton et al. , "Brown Adipose- tissue Mitochondria: Photoaffinity Labeling of the Regulatory Site of Energy Dissipation", J. Biochem. , Vol. 82, pp. 515-521 (1976) .
  • the activity of the uncoupling protein as a high conductance ion uniport through which H + ion can cross the mitochondrial membrane without ATP synthesis. Since brown adipose mitochondria are uncoupled due to the mitochondria uncoupling protein, the addition of nucleotides in-vitro will bind to the uncoupling protein thereby inhibiting the uncoupling of the mitochondria.
  • a competitive assay may be established by adding [ ⁇ "3 P] 8- azido-adenosine 5'triphosphate which is cross-linked to the active site of the uncoupling protein and will compete with GDP (guanosine 5'-diphosphate) for binding to the uncoupling protein.
  • the reduction in ATP counts will relate to the activity of the uncoupling protein in the sample.
  • the compositions prepared in accordance with the teachings of this invention are administered to a patient preferably by subcutaneous injection.
  • the dosage amount will vary over a wide range, given at appropriate intervals. It is preferred to provide the composition in a dose amount of at least 10 6 cells.
  • the desired dosage may be estimated by calculating the uncoupling activity of a given preparation of cells and adjusting for the desired activity.
  • the following Example is illustrative of the invention and is not intended to limit the invention as encompassed by the claims forming part of the application.
  • the pDR2 vector from Clontech Laboratories, Inc. with the gene for neomycin resistance is opened at a position in the polylinker and the cDNA for UCP is cloned in.
  • the plasmid is then selected for on ampicillin and grown up as a maxi-prep in the following manner.
  • the bacterial stock containing the plasmid is inoculated into 250ml of Luria-Bertrani broth containing 50ug/ml Ampicillin. This preparation is grown overnight at 37°C with vigorous shaking. The broth is centrifuged at 5,000 rpm for 10 minutes to pellet the bacteria. The resulting pellet is resuspended in 10ml of TRIS 50mM/EDTA lOmM (pH8) . 10ml of NaOH 200mM/l% SDS are added to the suspension and gently mixed by inversion. 10ml of 3M KHOAc (pH 5.5) is added to the mixture. The resulting preparation is mixed well and centrifuged at 5,000 rpm for 10 minutes.
  • the supernatant is precipitated in one volume of isopropanol and centrifuged at 10,000 rpm for 15 minutes.
  • the resulting pellet is resuspended in 5ml of water. 5g of solid cesium chloride is mixed to dissolve the suspended material.
  • 80ug of lOmg/ml of ethidium bromide is added to each one ml of DNA/cesium solution, mixed and placed in an ultracentrifuge tube.
  • the tube is topped off with light paraffin oil, sealed and loaded into an ultracentrifuge.
  • the ultracentrifuge is operated at 70,000 rpm overnight.
  • the lower closed circular band of DNA is obtained from the ultracentrifuge tube using a syringe.
  • An equal volume of water saturated with 1-butanol is added to the thus removed DNA band and mixed to remove the ethidium bromide.
  • the resulting mixture is centrifuged at 1500g to separate the phases.
  • the lower aqueous phase is transferred to a new tube and the extraction of ethidium bromide is repeated as necessary until the aqueous portion no longer has a pink color.
  • the DNA is then precipitated with ethanol, washed and resuspended in water to a concentration of one ug/ul.
  • the cells for transfection are trypsinized and 2.5 x 10 6 cells are added to a tube pelleted at 200g for 5 minutes and the supernatant is then aspirated. The cell pellet is then resuspended in the DNA precipitate prepared above and incubated for 15 minutes at room temperature.
  • DMEM + serum 9 ml of DMEM + serum is added to the tube and the contents of the tube are then transferred (5ml each) to two 100mm tissue culture plates.
  • the culture medium is changed every 24 hours.
  • Cells are suitable as transients 48 hours after transfection or may be selected for stable expression.
  • the presence of the gene product can be verified by ELISA or Western Blot.
  • the selected cells now express the gene for mitochondrial uncoupling protein. These cells are trypsinized and washed and resuspended in saline for injection directly into a subcutaneous location in the patient.
  • these cells may also be mixed with an isotonic solution of collagen and injected into a subcutaneous location in the patient.
  • Leu Arg lie Gly Leu Tyr Asp Thr Val Gin Glu Phe Leu Thr Ala
  • Met Arg Ser Val lie lie Asn Cys Thr Glu Leu Val Thr Tyr Asp

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Abstract

L'invention concerne un procédé servant à réaliser une perte de poids par utilisation de la protéine découplante mitochondriale, thermogénine, afin de convertir des calories en chaleur dans le tissu adipeux brun, ce qui provoque une diminution de poids due à une consommation d'énergie plus importante.
PCT/US1994/009799 1993-08-30 1994-08-29 Procede et composition de perte de poids WO1995006411A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU77178/94A AU7717894A (en) 1993-08-30 1994-08-29 Method and composition for weight reduction

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US11431393A 1993-08-30 1993-08-30
US08/114,313 1993-08-30
US24917994A 1994-05-25 1994-05-25
US08/249,179 1994-05-25

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WO1995006411A1 true WO1995006411A1 (fr) 1995-03-09

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998039432A1 (fr) * 1997-03-05 1998-09-11 Smithkline Beecham Plc Polynucleotides et polypeptides appartenant a la famille des proteines non liantes
GB2327224A (en) * 1997-07-09 1999-01-20 Mayo Foundation Gene transfer to adipocytes
WO2002007754A3 (fr) * 2000-07-25 2003-06-05 Smithkline Beecham Plc Nouvel usage
WO2003037362A3 (fr) * 2001-10-29 2003-12-24 Develogen Ag Proteines homologues de mnk kinase impliquees dans la regulation de l'homeostase energetique et du metabolisme des organites
WO2003002137A3 (fr) * 2001-06-27 2004-01-29 Develogen Ag Proteines homologues trp1, mct ou ftz-f1 impliquees dans la regulation de homeostasie energetique
WO2003040296A3 (fr) * 2001-11-08 2004-03-25 Develogen Ag Proteine men, gst2, rab-rp1, csp, proteine a f-box lilina/fbl7, abc50, coronine, sec61 alpha ou vhappa1-1 ou proteines homologues jouant un role dans la regulation de l'homeostasie energetique
US9044606B2 (en) 2010-01-22 2015-06-02 Ethicon Endo-Surgery, Inc. Methods and devices for activating brown adipose tissue using electrical energy
US10080884B2 (en) 2014-12-29 2018-09-25 Ethicon Llc Methods and devices for activating brown adipose tissue using electrical energy
US10092738B2 (en) 2014-12-29 2018-10-09 Ethicon Llc Methods and devices for inhibiting nerves when activating brown adipose tissue

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, Volume 167, Number 2, issued 16 March 1990, S. KLAUS et al., "Expression of the Brown Fat Mitochondria Uncoupling Protein in Xenopus Oocytes and Import Into Mitochondrial Membrane", pages 784-789. *
D.B. DILL et al., "Handbook of Physiology. Adipose Tissue", Published 1964, by OXFORD UNIVERSITY PRESS, (N.Y.), pages 59-85, Chapter 9. *
NUCLEIC ACIDS RESEARCH, Volume 17, Number 5, issued 1989, L. CASTEILLA et al., "Nucleotide Sequence of a cDNA Encoding Bovine Brown Fat Uncoupling Protein. Homology With ADP Binding Site of ADP/ATP Carrier", page 2131. *
PROC. NATL. ACAD. SCI. U.S.A., Volume 87, issued July 1990, L. CASTEILLA et al., "Stable Expression of Functional Mitochondrial Uncoupling Protein in Chinese Hamster Ovary Cells", pages 5124-5128. *
THE JOURNAL OF BIOLOGICAL CHEMISTRY, Volume 260, Number 30, issued 25 December 1985, A. JACOBSSON et al., "Mitochondrial Uncoupling Protein From Mouse Brown Fat. Molecular Cloning, Genetic Mapping and mRNA Expression", pages 16250-16254. *
THE JOURNAL OF BIOLOGICAL CHEMISTRY, Volume 263, Number 25, issued 05 September 1988, L.P. KOZAK et al., "The Mitochondrial Uncoupling Protein Gene", pages 12274-12277. *

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998039432A1 (fr) * 1997-03-05 1998-09-11 Smithkline Beecham Plc Polynucleotides et polypeptides appartenant a la famille des proteines non liantes
GB2327224A (en) * 1997-07-09 1999-01-20 Mayo Foundation Gene transfer to adipocytes
WO2002007754A3 (fr) * 2000-07-25 2003-06-05 Smithkline Beecham Plc Nouvel usage
WO2003002137A3 (fr) * 2001-06-27 2004-01-29 Develogen Ag Proteines homologues trp1, mct ou ftz-f1 impliquees dans la regulation de homeostasie energetique
WO2003037362A3 (fr) * 2001-10-29 2003-12-24 Develogen Ag Proteines homologues de mnk kinase impliquees dans la regulation de l'homeostase energetique et du metabolisme des organites
US8076098B2 (en) 2001-10-29 2011-12-13 Boehringer Ingelheim International Gmbh Mnk kinase homologous proteins involved in the regulation of energy homeostasis and organelle metabolism
US8828934B2 (en) 2001-10-29 2014-09-09 Boehringer Ingelheim International Gmbh Mnk kinase homologous proteins involved in the regulation of energy homeostasis and organelle metabolism
US8957020B2 (en) 2001-10-29 2015-02-17 Boehringer Ingelheim International Gmbh Mnk kinase homologous proteins involved in the regulation of energy homeostasis and organelle metabolism
WO2003040296A3 (fr) * 2001-11-08 2004-03-25 Develogen Ag Proteine men, gst2, rab-rp1, csp, proteine a f-box lilina/fbl7, abc50, coronine, sec61 alpha ou vhappa1-1 ou proteines homologues jouant un role dans la regulation de l'homeostasie energetique
US9662486B2 (en) 2010-01-22 2017-05-30 Ethicon Endo-Surgery, Inc. Methods and devices for activating brown adipose tissue using electrical energy
US9044606B2 (en) 2010-01-22 2015-06-02 Ethicon Endo-Surgery, Inc. Methods and devices for activating brown adipose tissue using electrical energy
US10201695B2 (en) 2010-01-22 2019-02-12 Ethicon Endo-Surgery, Inc. Methods and devices for activating brown adipose tissue using electrical energy
US11040196B2 (en) 2010-01-22 2021-06-22 Cilag Gmbh International Methods and devices for activating brown adipose tissue using electrical energy
US10080884B2 (en) 2014-12-29 2018-09-25 Ethicon Llc Methods and devices for activating brown adipose tissue using electrical energy
US10092738B2 (en) 2014-12-29 2018-10-09 Ethicon Llc Methods and devices for inhibiting nerves when activating brown adipose tissue
US10207102B2 (en) 2014-12-29 2019-02-19 Ethicon Llc Methods and devices for activating brown adipose tissue using electrical energy
US10391298B2 (en) 2014-12-29 2019-08-27 Ethicon Llc Methods and devices for activating brown adipose tissue using electrical energy
US10960201B2 (en) 2014-12-29 2021-03-30 Ethicon Llc Methods and devices for inhibiting nerves when activating brown adipose tissue
US10994123B2 (en) 2014-12-29 2021-05-04 Cilag Gmbh International Methods and devices for activating brown adipose tissue using electrical energy
US11679252B2 (en) 2014-12-29 2023-06-20 Cilag Gmbh International Methods and devices for activating brown adipose tissue using electrical energy

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