[go: up one dir, main page]

WO2025045915A1 - Procédés pour induire une hypertrophie musculaire - Google Patents

Procédés pour induire une hypertrophie musculaire Download PDF

Info

Publication number
WO2025045915A1
WO2025045915A1 PCT/EP2024/074017 EP2024074017W WO2025045915A1 WO 2025045915 A1 WO2025045915 A1 WO 2025045915A1 EP 2024074017 W EP2024074017 W EP 2024074017W WO 2025045915 A1 WO2025045915 A1 WO 2025045915A1
Authority
WO
WIPO (PCT)
Prior art keywords
pik3ca
protein
agent
fragment
protein expression
Prior art date
Legal status (The legal status 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 status listed.)
Pending
Application number
PCT/EP2024/074017
Other languages
English (en)
Inventor
Guillaume CANAUD
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Centre National de la Recherche Scientifique CNRS
Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Cite
Original Assignee
Centre National de la Recherche Scientifique CNRS
Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Cite
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 Centre National de la Recherche Scientifique CNRS, Assistance Publique Hopitaux de Paris APHP, Institut National de la Sante et de la Recherche Medicale INSERM, Universite Paris Cite filed Critical Centre National de la Recherche Scientifique CNRS
Publication of WO2025045915A1 publication Critical patent/WO2025045915A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/473—Quinolines; Isoquinolines ortho- or peri-condensed with carbocyclic ring systems, e.g. acridines, phenanthridines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • 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
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/45—Transferases (2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system

Definitions

  • the present invention is in the field of medicine, in particular rheumatology.
  • Muscular atrophy refers to a reduction in muscle mass, more particularly a reduction in striated skeletal muscle. The causes of this atrophy can be varied. Muscular atrophy is observed in pathological situations (e.g. neuromuscular diseases, metabolic disorders, hypercatabolism, eating disorders, cachexia, sarcopenia, medication) as well as physiological situations (e.g. ageing, lack of physical activity, malnutrition). Whatever the case, muscular atrophy can be deleterious leading to weakness, fatigue and reduced quality of life, as well as frailty and an increased risk of falls, fractures and loss of independence.
  • pathological situations e.g. neuromuscular diseases, metabolic disorders, hypercatabolism, eating disorders, cachexia, sarcopenia, medication
  • physiological situations e.g. ageing, lack of physical activity, malnutrition.
  • muscular atrophy can be deleterious leading to weakness, fatigue and reduced quality of life, as well as frailty and an increased risk of falls, fractures and loss of independence.
  • Hemifacial myohyperplasia is a rare cause of facial asymmetry exclusively involving facial muscles, initially reported as ‘hypertrophy and asymmetry of the facial muscles’ 1 . This disorder is reported in very few patients in the literature 2 ' 6 .
  • the clinical presentation of HFMH patients is strikingly consistent, with unilateral muscular hypertrophy mimicking spasm and orofacial dystonia, leading to diagnostic errors and inadequate management strategies, including aggressive attempts of surgical correction 6 .
  • the genetic causes of HFMH are currently unknown.
  • the recent discovery of the role played by somatic mutation of genes activating the PIK3CA/AKT/mTOR pathway has opened new treatment perspectives for patients 7 .
  • PIK3CA gain-of-function mutations explain the vast majority of overgrowth syndromes 7 .
  • PIK3CA encodes the 110-Kd catalytic a-subunit of PI3K, a lipid kinase that controls signaling pathways involved in cell proliferation, motility, survival and metabolism 8 .
  • Post-zygotic mosaic gain-of-function PIK3CA mutations result in protein activation, leading to abnormal cellular proliferation, tissue hyperplasia and organ overgrowth.
  • the present invention relates to a PIK3CA protein or fragment thereof and/or an agent for PIK3CA protein expression for use in promoting skeletal muscle hypertrophy in a subject in need thereof.
  • PIK3CA/AKT/mTOR pathway was abnormally affected in patients with HFMH. They identified a somatic gain-of-function mutation of PIK3CA in 5 pediatric patients with hemifacial myohyperplasia, a rare cause of facial asymmetry exclusively involving facial muscles. To understand the physiopathology of muscle hypertrophy, they created a mouse model carrying specifically a PIK3CA mutation in skeletal muscles. PIK3CA gain-of-function mutation led to striated muscle cell hypertrophy, mitochondria dysfunction and hypoglycemia with low circulating insulin levels. A PIK3CA inhibitor, namely Alpelisib, was able to prevent and to reduce muscle hypertrophy in the mouse model. They then concluded that PIK3CA is a relevant target to induce skeletal muscle hypertrophy.
  • the present invention relates to a PI3K protein or fragment thereof and/or an agent for PI3K protein expression for use in promoting skeletal muscle hypertrophy in a subject in need thereof. More particularly, the present invention relates to a PIK3CA protein or fragment thereof and/or an agent for PIK3CA protein expression for use in promoting skeletal muscle hypertrophy in a subject in need thereof.
  • the term “subject” denotes a vertebrate such as mammal, bird, fish, amphibian or reptile.
  • the vertebrate is a warm-blooded vertebrate (i.e. mammal, bird, fish).
  • the subject is a mammal, such as a rodent, a feline, a canine (e.g. a dog), an equine (e.g. horse), a bovine (e.g. a beef), a sheep or a primate. More particularly, the subject according to the invention is a human. In some embodiments, the subject suffers from muscular atrophy, muscular dystrophy or muscular hypotrophy.
  • the subject suffers from muscle wasting, decreased muscle tone, muscle weakness or muscle fragility. In some embodiments, the subject suffers from sarcopenia (i.e. gradual loss of skeletal muscle mass and strength usually associated with advanced ageing). In some embodiments, the subject suffers from cachexia (i.e. multifactorial syndrome with involuntary progressive weight loss characterized by skeletal muscle loss). In some embodiments, the subject suffers from overweight or obesity (i.e. respectively body mass index over 25 or 30).
  • skeletal muscle hypertrophy or “skeletal muscle development” refers to an increase in skeletal muscle fiber cross sectional area and skeletal muscle mass.
  • the invention also relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression for use in promoting skeletal muscle hypertrophy in a subject suffering from muscular atrophy, muscular dystrophy or muscular hypotrophy.
  • the invention also relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression for use in promoting skeletal muscle hypertrophy in a subject suffering from muscle wasting, decreased muscle tone, muscle weakness or muscle fragility.
  • the invention also relates to the PIK3CA protein or fragment thereof and/or the agent for PIKCA protein expression for use in the treatment of sarcopenia in a subject in need thereof.
  • the invention also relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression for use in the treatment of cachexia in a subject in need thereof.
  • treatment refers to both prophylactic or preventive treatment as well as curative or disease-modifying treatment, including treatment of subjects at risk of contracting the disease or suspected to have contracted the disease as well as subjects who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
  • the treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
  • therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
  • a therapeutic regimen may include an induction regimen and a maintenance regimen.
  • the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
  • the general goal of an induction regimen is to provide a high level of drug to a subject during the initial period of a treatment regimen.
  • An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
  • maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a subject during treatment of an illness, e.g., to keep the subject in remission for long periods of time (months or years).
  • a maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
  • PI3K refers to phosphoinositide 3-kinases also called phophatidylinositide 3-kinases.
  • PI3K belongs to a family of enzymes which phosphorylate the 3 ’hydroxyl group of the inositol ring of the phosphatidylinositol (Ptdins).
  • Ptdins phosphatidylinositol
  • the PI3K signalling pathway can be activated, resulting in the synthesis of PIP3 from PIP2.
  • PIK3CA is mainly recruited through tyrosine kinase receptors.
  • PIK3CA refers to a protein encoded by the PIK3CA gene (Entrez Gene: 5290; Ensembl: ENSG00000121879).
  • PIK3CA gene encodes the 110-kDa catalytic alpha subunit of PI3K (pl 10a), which converts, at the plasma membrane, phosphatidylinositol 4, 5 -bisphosphate (PtdIns(4,5)P2) to phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3; or PIP3) with subsequent recruitment of PDK1, which in turn phosphorylates AKT on the Thr308 residue to initiate downstream cellular effects.
  • PIK3CA also regulates many other pathways, including the Rho/Racl signaling cascade.
  • SEQ ID NO:1 An exemplary amino acid sequence for PIK3CA is shown as SEQ ID NO:1.
  • an agent for PIK3CA protein expression denotes a molecule that partially or totally enhances PIK3CA biological activity (e.g. kinase activity) or expression. The term also encompasses a molecule able to increase or restore PIK3CA gene expression.
  • a test is necessary. For that purpose, to identify agent for PIK3CA protein expression, a western blot analysis can be run on cell extracts to test the effect on the putative agent for PIK3CA protein expression on the level of PIK3CA. If the level of PIK3CA is increased, the putative agent for PIK3CA protein expression will have the desired effect.
  • the agent for PIK3CA protein expression according to the invention is a low molecular weight compound, e. g. a small organic molecule (natural or not).
  • the small organic molecule is a PIK3CA agonist.
  • the small organic molecule is an activator of PIK3CA, in particular an allosteric activator of PIK3CA.
  • small organic molecule refers to a molecule (natural or not) of a size comparable to those organic molecules generally used in pharmaceuticals. The term excludes biological macromolecules (e. g., proteins, nucleic acids, etc.).
  • PLOS ONE 9(1): e84964) or aminopyridines such as l-(7-((2-((4-(4-ethylpiperazin-l- yl)phenyl)amino)pyridin-4-yl)amino)indolin-l-yl)ethan-l-one (“UCL-TRO-1938”, “1938”; CAS N°: 2919575-27-0; Gong, Grace Q et al. “A small-molecule PI3Ka activator for cardioprotection and neuroregeneration.” Nature vol. 618,7963 (2023): 159-168). Other agents for PIK3CA protein expression are described in W02023/041905.
  • a first amino acid sequence having at least 70% of identity with a second amino acid sequence means that the first sequence has 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80; 81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 98; or 99, or 100% of identity with the second amino acid sequence.
  • Amino acid sequence identity is preferably determined using a suitable sequence alignment algorithm and default parameters, such as BLAST P (Karlin and Altschul, 1990).
  • the PIK3CA protein of the invention is a functional conservative variant of the PIK3CA protein according to the invention.
  • a “function-conservative variant” are those in which a given amino acid residue in a protein or enzyme has been changed without altering the overall conformation and function of the PIK3CA protein, including, but not limited to, replacement of an amino acid with one having similar properties (such as, for example, polarity, hydrogen bonding potential, acidic, basic, hydrophobic, aromatic, and the like). Accordingly, a “functionconservative variant” also includes an PIK3CA protein which has at least 70 % amino acid identity and which has the same or substantially similar properties or functions as the native or parent PIK3CA protein to which it is compared. Functional properties of the PIK3CA protein of the invention could typically be assessed in any functional assay as described in the EXAMPLE.
  • a further aspect of the present invention relates to a fusion protein comprising the PIK3CA protein or peptide according to the invention that is fused to at least one heterologous polypeptide.
  • fusion protein refers to the PIK3CA protein or peptide according to the invention that is fused directly or via a spacer to at least one heterologous polypeptide.
  • the fusion protein comprises the protein or peptide according to the invention that is fused either directly or via a spacer at its C-terminal end to the N-terminal end of the heterologous polypeptide, or at its N-terminal end to the C-terminal end of the heterologous polypeptide.
  • the term “directly” means that the (first or last) amino acid at the terminal end (N or C-terminal end) of the protein or peptide is fused to the (first or last) amino acid at the terminal end (N or C-terminal end) of the heterologous polypeptide.
  • the last amino acid of the C-terminal end of said protein or peptide is directly linked by a covalent bond to the first amino acid of the N- terminal end of said heterologous polypeptide, or the first amino acid of the N-terminal end of said protein or peptide is directly linked by a covalent bond to the last amino acid of the C- terminal end of said heterologous polypeptide.
  • cell-penetrating peptide is well known in the art and refers to a cell permeable sequence or membranous penetrating sequence such as penetratin, TAT mitochondrial penetrating sequence and compounds (Bechara and Sagan, 2013; Jones and Sayers, 2012; Khafagy el and Morishita, 2012; Malhi and Murthy, 2012).
  • the proteins, peptides or fusion proteins of the invention may be produced by any technique known per se in the art, such as, without limitation, any chemical, biological, genetic or enzymatic technique, either alone or in combination. Knowing the amino acid sequence of the desired sequence, one skilled in the art can readily produce said proteins, peptides or fusion proteins, by standard techniques for production of amino acid sequences. For instance, they can be synthesized using well-known solid phase method, preferably using a commercially available peptide synthesis apparatus (such as that made by Applied Biosystems, Foster City, California) and following the manufacturer’s instructions. Alternatively, the proteins, peptides or fusion proteins of the invention can be synthesized by recombinant DNA techniques as is now well-known in the art.
  • these fragments can be obtained as DNA expression products after incorporation of DNA sequences encoding the desired (poly) peptide into expression vectors and introduction of such vectors into suitable eukaryotic or prokaryotic hosts that will express the desired (poly) peptide, from which they can be later isolated using well- known techniques.
  • the proteins, peptides or fusion proteins of the invention can be used in an isolated (e.g., purified) form or contained in a vector, such as a membrane or lipid vesicle (e.g. a liposome).
  • proteins, peptides or fusion proteins according to the invention may be modified in order to improve their therapeutic efficacy and their stability using well-known techniques.
  • modification of therapeutic compounds may be used to decrease toxicity, increase circulatory time, or modify biodistribution.
  • the toxicity of potentially important therapeutic compounds can be decreased significantly by combination with a variety of drug carrier vehicles that modify biodistribution.
  • a strategy for improving drug stability is the utilization of water-soluble polymers.
  • Various water-soluble polymers have been shown to modify biodistribution, improve the mode of cellular uptake, change the permeability through physiological barriers; and modify the rate of clearance from the body.
  • water-soluble polymers have been synthesized that contain drug moieties as terminal groups, as part of the backbone, or as pendent groups on the polymer chain.
  • Pegylation is a well-established and validated approach for the modification of a range of polypeptides (Chapman, 2002).
  • the benefits include among others: (a) markedly improved circulating half-lives in vivo due to either evasion of renal clearance as a result of the polymer increasing the apparent size of the molecule to above the glomerular filtration limit, and/or through evasion of cellular clearance mechanisms; (b) reduced antigenicity and immunogenicity of the molecule to which PEG is attached; (c) improved pharmacokinetics; (d) enhanced proteolytic resistance of the conjugated protein (Cunningham-Rundles et.al., 1992); and (e) improved thermal and mechanical stability of the PEGylated polypeptide.
  • the proteins, peptides or fusion proteins of the invention may be covalently linked with one or more polyethylene glycol (PEG) group(s).
  • PEG polyethylene glycol
  • One skilled in the art can select a suitable molecular mass for PEG, based on how the pegylated polypeptide will be used therapeutically by considering different factors including desired dosage, circulation time, resistance to proteolysis, immunogenicity, etc.
  • the PEG of the invention terminates on one end with hydroxy or methoxy, i.e., X is H or CH3 ("methoxy PEG").
  • X is H or CH3
  • such a PEG can consist of one or more PEG sidechains which are linked together. PEGs with more than one PEG chain are called branched PEGs.
  • Branched PEGs can be prepared, for example, by the addition of polyethylene oxide to various polyols, including glycerol, pentaerythriol, and sorbitol.
  • a four-armed branched PEG can be prepared from pentaerythriol and ethylene oxide.
  • One form of PEGs includes two PEG side-chains (PEG2) linked via the primary amino groups of a lysine (Monfardini et al., 1995).
  • PEG2 PEG side-chains
  • the hydroxyl end groups of the polymer molecule must be provided in activated form, i. e.
  • Suitable activated polymer molecules are commercially available, e. g. from Shearwater Polymers, Inc., Huntsville, AL, USA, or from PolyMASC Pharmaceuticals pic, UK.
  • the polymer molecules can be activated by conventional methods known in the art, e. g. as disclosed in WO 90/13540.
  • activated linear or branched polymer molecules for use in the present invention are described in the Shearwater Polymers, Inc. 1997 and 2000 Catalogs (Functionalized Biocompatible Polymers for Research and pharmaceuticals, Polyethylene Glycol and Derivatives, incorporated herein by reference).
  • activated PEG polymers include the following linear PEGs : NHS-PEG (e.g.
  • SPA-PEG SSPA-PEG, SBA-PEG, SS-PEG, SSA-PEG, SC-PEG, SG-PEG, and SCM- PEG
  • NOR-PEG BTC-PEG, EPOX-PEG, NCO-PEG, NPC-PEG, CDI-PEG, ALD-PEG, TRES-PEG, VS-PEG, IODO-PEG, and MAL-PEG, and branched PEGs such as PEG2-NHS.
  • the conjugation of the proteins, peptides or fusion proteins and the activated polymer molecules is conducted by use of any conventional method. Conventional methods are known to the skilled artisan. The skilled person will be aware that the activation method and/or conjugation chemistry to be used depends on the attachment group(s) of the polypeptides as well as the functional groups of the PEG molecule (e.g., being amine, hydroxyl, carboxyl, aldehyde, ketone, sulfhydryl, succinimidyl, maleimide, vinylsulfone or haloacetate).
  • the attachment group(s) of the polypeptides as well as the functional groups of the PEG molecule (e.g., being amine, hydroxyl, carboxyl, aldehyde, ketone, sulfhydryl, succinimidyl, maleimide, vinylsulfone or haloacetate).
  • the proteins, peptides or fusion proteins of the invention are conjugated with PEGs at amino acid D and E (for COOH), T, Y and S (for OH), K (for NH2), C (for SH if at least one cysteine is conserved) or/and Q and N (for the amide function).
  • additional sites for PEGylation can be introduced by site-directed mutagenesis by introducing one or more lysine residues. For instance, one or more arginine residues may be mutated to a lysine residue.
  • additional PEGylation sites are chemically introduced by modifying amino acids on proteins, peptides or fusion proteins of the invention.
  • PEGs are conjugated to the polypeptides or fusion proteins through a linker.
  • Suitable linkers are well known to the skilled person.
  • a preferred example is cyanuric chloride ((Abuchowski et al., 1977); US 4,179, 337).
  • Conventional separation and purification techniques known in the art can be used to purify pegylated polypeptides of the invention, such as size exclusion (e.g. gel filtration) and ion exchange chromatography. Products may also be separated using SDS- PAGE.
  • the pegylated polypeptides provided by the invention have a serum half-life in vivo at least 50%, 75%, 100%, 150% or 200% greater than that of an unmodified polypeptide.
  • the agent for PIK3CA protein expression of the invention is selected from the group consisting of an isolated, synthetic or recombinant nucleic acid encoding for PIK3CA protein, a nucleic acid sequence encoding for the fusion protein, a nucleic acid encoding a fragment of a PIK3CA protein, a nucleic acid encoding a fragment of a peptide according to the invention, a cell expressing PIK3CA protein, and/or an agent inducing PIK3CA gene expression and their combinations.
  • the agent for PIK3CA protein expression of the invention is a polynucleotide encoding a polypeptide that comprises the amino acid sequence as set forth in SEQ ID NO:1.
  • the agent for PIK3CA protein expression of the invention is a polynucleotide encoding a polypeptide having at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 % identity with the amino acid sequence set forth in SEQ ID NO:1.
  • a sequence "encoding" an expression product such as a RNA, polypeptide, protein, or enzyme
  • a sequence "encoding" an expression product is a nucleotide sequence that, when expressed, results in the production of that RNA, polypeptide, protein, or enzyme, i.e., the nucleotide sequence encodes an amino acid sequence for that polypeptide, protein or enzyme.
  • a coding sequence for a protein may include a start codon (usually ATG) and a stop codon.
  • said nucleic acid is a DNA or RNA molecule, which may be included in a suitable vector, such as a plasmid, cosmid, episome, artificial chromosome, phage or viral vector.
  • a further object of the present invention relates to a vector and an expression cassette in which a nucleic acid molecule encoding for a proteins, peptides or fusion proteins of the invention is associated with suitable elements for controlling transcription (in particular promoter, enhancer and, optionally, terminator) and, optionally translation, and also the recombinant vectors into which a nucleic acid molecule in accordance with the invention is inserted.
  • recombinant vectors may, for example, be cloning vectors, or expression vectors.
  • vector means the vehicle by which a DNA or RNA sequence (e.g. a foreign gene) can be introduced into a host cell, so as to transform the host and promote expression (e.g. transcription and translation) of the introduced sequence.
  • a DNA or RNA sequence e.g. a foreign gene
  • Any expression vector for animal cell can be used.
  • suitable vectors include pAGE107 (Miyaji et al., 1990), pAGE103 (Mizukami and Itoh, 1987), pHSG274 (Brady et al., 1984), pKCR (O'Hare et al., 1981), pSGl beta d2-4 (Miyaji et al., 1990) and the like.
  • Plasmids include replicating plasmids comprising an origin of replication, or integrative plasmids, such as for instance pUC, pcDNA, pBR, and the like.
  • viral vectors include adenoviral, lentiviral, retroviral, herpes virus and AAV vectors.
  • recombinant viruses may be produced by techniques known in the art, such as by transfecting packaging cells or by transient transfection with helper plasmids or viruses.
  • virus packaging cells include PA317 cells, PsiCRIP cells, GPenv+ cells, 293 cells, etc.
  • promoters and enhancers used in the expression vector for animal cell include early promoter and enhancer of SV40 (Mizukami and Itoh, 1987), LTR promoter and enhancer of Moloney mouse leukemia virus (Kuwana et al., 1987), promoter (Mason et al., 1985) and enhancer (Gillies et al., 1983) of immunoglobulin H chain and the like.
  • a further aspect of the invention relates to a host cell comprising a nucleic acid molecule encoding for a protein, peptide or a fusion protein according to the invention or a vector according to the invention.
  • a subject of the present invention is a prokaryotic or eukaryotic host cell genetically transformed with at least one nucleic acid molecule or vector according to the invention.
  • transformation means the introduction of a "foreign” (i.e. extrinsic or extracellular) gene, DNA or RNA sequence to a host cell, so that the host cell will express the introduced gene or sequence to produce a desired substance, typically a protein or enzyme coded by the introduced gene or sequence.
  • a host cell that receives and expresses introduced DNA or RNA has been "transformed".
  • prokaryotic cells in particular E. coli cells
  • prokaryotic cells have the advantages to produce protein in large amounts. If a eukaryotic context is needed, yeasts (e.g. saccharomyces strains) may be particularly suitable since they allow production of large amounts of proteins.
  • the protein, peptide or the fusion protein of the invention can, for example, be obtained by culturing genetically transformed cells in accordance with the invention and recovering the proteins, peptides or fusion proteins expressed by said cell, from the culture. They may then, if necessary, be purified by conventional procedures, known in themselves to those skilled in the art, for example by fractional precipitation, in particular ammonium sulfate precipitation, electrophoresis, gel filtration, affinity chromatography, etc. In particular, conventional methods for preparing and purifying recombinant proteins may be used for producing the proteins in accordance with the invention.
  • the present invention relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression according to the invention in combination with one or more compound promoting skeletal muscle hypertrophy (e.g. steroids).
  • one or more compound promoting skeletal muscle hypertrophy e.g. steroids.
  • the present invention relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression according to the invention in combination with one or more compound promoting skeletal muscle hypertrophy for use in the treatment of sarcopenia in a subject in need thereof.
  • the present invention relates to the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression according to the invention in combination with one or more compound promoting skeletal muscle hypertrophy for use in the treatment of cachexia in a subject in need thereof.
  • the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression according to the invention as described above are administered to the subject in a therapeutically effective amount.
  • a therapeutically effective amount of the PIK3CA protein or fragment thereof and/or of the agent for PIK3CA protein expression of the present invention as above described is meant a sufficient amount of the PIK3CA protein or fragment thereof and/or of the agent for PIK3CA protein expression for treating the disease at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood, however, that the total daily usage of the PIK3CA protein or fragment thereof and/or an agent for PIK3CA protein expression of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific the PIK3CA protein or fragment thereof and/or of the agent for PIK3CA protein expression employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the PIK3CA protein or fragment thereof and/or of the agent for PIK3CA protein expression employed; the duration of the treatment; drugs used in combination or coincidental with the PIK3CA protein or fragment thereof and/or with the agent for PIK3CA protein expression employed; and like factors well known in the medical arts.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the PIK3CA protein or fragment thereof and/or of the agent for PIK3CA protein expression of the present invention for the symptomatic adjustment of the dosage to the subject to be treated.
  • the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression according to the invention may be used in a concentration between 0.01 pM and 20 pM, particularly, the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression of the invention may be used in a concentration of 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 20.0 pM.
  • the PIK3C A protein or fragment thereof and/or the agent for PIK3CA protein expression of the present invention is administered to the subject in the form of a pharmaceutical composition.
  • the invention also relates to a therapeutic composition comprising the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression for use in promoting skeletal muscle hypertrophy in a subject in need thereof.
  • the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression of the present invention may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions.
  • pharmaceutically acceptable excipients such as a carboxylate, ethylene glycol, ethylene glycol, ethylene glycol, ethylene glycol, ethylene glycol, ethylene glycol, kaolin, kaolin, kaolin, kaolin, kaolin, kaolin, kaolin, adiluent, encapsulating material or formulation auxiliary of any type.
  • the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • sterile powders for the preparation of sterile injectable solutions the typical methods of preparation are vacuumdrying and freeze-drying techniques which yield a powder of the PIK3CA protein or fragment thereof and/or the agent for PIK3CA protein expression of the present invention plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • the preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents to a small tumor area.
  • solutions Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.
  • FIGURES are a diagrammatic representation of FIGURES.
  • Figure 1 A mouse model of PZ 3C4-related skeletal muscle overgrowth.
  • B. Adipose tissue and skeletal muscle volume quantification.
  • C. Strength measured using grip test (n 3 per group).
  • D. Representative pictures of PIK3CA WT and PIK3CA HSA ⁇ CreER mice.
  • E Western blot of pl 10a and GFP and P-AKT Ser473 in skeletal muscles of PIK3CA WT and PIK3CA HSA ⁇ CreER mice.
  • FIG. 2 Mouse model characterization and quantitative histological analysis of tibialis anterior (TA) muscle changes in PIK3CA WT and PIK3CA HSA ' CreER mice treated with either vehicle or alpelisib.
  • A Representative immunofluorescence of PAX7 satellite cells and quantification.
  • B Quantification of number of fibers per muscle and TA muscle cross section area (CSA) in PIK3CA WT and PIK3CA HSA ⁇ CreER mice treated with either vehicle or alpelisib.
  • C Complete blood count in PIK3CA WT and PIK3CA HSA ⁇ CreER mice.
  • D p Islet area in in PIK3CA WT and PIK3CA HSA ⁇ CreER mice.
  • FIG. 3 Alpelisib prevents and reverses skeletal muscle overgrowth in PIK3CA HSA ' CreER mice.
  • the pl 10* protein expressed by R26StopFLP 110* mice is a constitutively active chimera that contains the iSH2 domain of p85 fused to the NH2- terminus of pl 10 via a flexible glycine linker 11 .
  • a cloned loxP- flanked neoR-stop cassette was inserted into a modified version of pROSA26-l, followed by cDNA encoding pl 10* and then a frt-flanked IRES-EGFP cassette and a bovine polyadenylation sequence (R26StopFLP 170*) 12 .
  • mice were then interbred with G t (ROSA)26SoP m4(ACTBAdTomato ’- EGFP)Luo/J mice 10,13 . These mice express a cell membrane-localized tdTomato fluorescent protein in all tissues that is replaced by GFP after Cre recombination. Animals were fed ad libitum and housed at a constant ambient temperature in a 12-h light cycle. Animals were fed with regular chow food (2018 Teklad global 18% protein rodent diets, 3.1 Kcal/g, Envigo).
  • mice [ and PIK3CA HSA ⁇ CreER mice were treated with the PIK3CA inhibitor alpelisib (MedChem Express, Germany; 50 mg.kg-1 in 0.5% carboxymethylcellulose (Sigma Aldrich), daily p.o.) or vehicle (0.5% carboxymethylcellulose (Sigma Aldrich), daily p.o.). Treatment was started either immediately (preventive study) or 6 weeks (therapeutic study) following Cre induction. The last dose of alpelisib or vehicle was administered approximately 3 hours before sacrifice. All mice were fasted for 12 hours before blood glucose measurement (Accuchek Performa, Roche Diagnostic), Magnetic Resonance Imaging and sacrifice.
  • alpelisib MedChem Express, Germany; 50 mg.kg-1 in 0.5% carboxymethylcellulose (Sigma Aldrich), daily p.o.) or vehicle (0.5% carboxymethylcellulose (Sigma Aldrich), daily p.o.). Treatment was started either immediately (preventive study) or 6 weeks (therapeutic study) following Cre induction. The last dose of al
  • mice Grip strength performance of the mice was evaluated using a grip strength dynamometer obtained from Bioseb (Chaville, France). To assess hindlimb muscle strength, the mice were positioned on a grid surface. Gentle traction was applied to the mice's tails in the opposite direction, and the maximum strength exerted by each mouse before releasing their grip was measured five times. A recovery period of 30 seconds was provided between each measurement. The average value from the five measurements was calculated as the indicator of hindlimb grip strength. Magnetic Resonance Imaging (MRI) evaluation
  • MRI Magnetic Resonance Imaging
  • mice were fasted overnight with free access to water. Mice were then anesthetized (2 ⁇ 0.5% isoflurane in dioxygen) and weighed, and glycemia was measured in blood drawn from the caudal ventral artery using an Accu-Chek® Aviva Nano A (Accu-Chek, France).
  • a 29G needle catheter (Fischer Scientific, France) connected to 5 cm polyethylene tubing (Tygon Microbore Tubing, 0.010" x 0.030"OD; Fisher Scientific, France) was inserted into the caudal vein for radiotracer injection.
  • CT projections were reconstructed by filtered retroproj ection (filter: cosine; cutoff: 100%) using Nucline 3.00.010.0000 software (Mediso Medical Imaging Systems, Hungary). Fifty-five minutes post tracer injection, PET data were collected for 10 min in list mode and binned using a 5 ns time window, with a 400-600 keV energy window and a 1 :5 coincidence mode. Data were reconstructed using the Tera-Tomo reconstruction engine (3D-OSEM-based manufactured customized algorithm) with expectation maximization iterations, scatter, and attenuation correction.
  • 3D-OSEM-based manufactured customized algorithm the Tera-Tomo reconstruction engine
  • VIs Volumes of interest
  • PMOD Technologies Ltd Zurich, Switzerland
  • FDG accumulation was quantified as the standard uptake value (SUV), which measures the ratio of the radioactivity concentration in VOI to the whole-body concentration of the injected radioactivity.
  • SUV standard uptake value
  • mice blood samples were collected from the mice in EDTA- coated tubes.
  • fresh blood samples were analyzed on a hematology analyzer (ProCyte Dx; IDEXX Laboratories) and centrifuged at 500*g for 15 min.
  • the collected plasma concentration was used to determine insulin (U-PLEX mouse insulin Assay (Meso Scale Discovery) ref# K1526HK) and IGF-1 (Novus Biologicals, ref# MG100) circulating levels using enzymatic methods from commercially available kits.
  • mice were fasted overnight (12 h) with access to drinking water. All body weights were measured, and tails were carefully cut for blood glucose determination (time point 0).
  • GTT glucose tolerance test
  • freshly prepared glucose solution was administered orally (1 g glucose/kg body weight of 20% glucose solution in water, (Sigma, ref# G8270)). After 15, 30, 45, 60, 90 and 120 minutes, blood glucose was measured again at all time points.
  • Hematoxylin and eosin (H&E) slides were scanned with a NanoZoomer 2.0HT (Hamamatsu) and analyzed with Qupath-0.2.3 13 . Muscles were segmented using deep learning Cellpose algorithm 14 . For both, areas were then measured with Fiji 15 .
  • Paraffin-embedded tissue sections (4 pm) were submitted to antigen retrieval protocols using high temperature (120 °C) and high pressure in citrate buffer and a pressure cooker. Sections were then incubated with primary antibodies.
  • appropriate Al exafluor-conjugated secondary antibodies (Thermo Fischer Scientific) were incubated on the samples and analyzed using an LSM 700 confocal microscope (Zeiss) or Eclipse Ni-E (Nikon). Immunohistochemistry revelation was performed with appropriate horseradish peroxidase (HRP) linked secondary antibodies and analyzed with E800 (Nikon).
  • Tissues were crushed and then lysed in RIPA lysis buffer supplemented with phosphatase and protease inhibitors. Protein concentrations were determined through the bicinchoninic acid method (Pierce). Then, protein extracts were resolved by SDS-PAGE before being transferred onto the appropriate membrane and incubated with the primary antibody followed by the appropriate peroxidase-conjugated secondary antibody (dilution 1 : 10,000). Chemiluminescence was acquired using Chemidoc MP, and bands were quantitated using Image Lab Software (Bio-Rad Laboratories).
  • Muscles of PIK3CA WT and PIK3CA HSA ⁇ CreER mice were harvested and rinsed with PBS IX (Gibco). After cutting them into small pieces, digestion buffer was added with DNAse (0,lmg/mL), Dispase I (0,8mg/mL), Collagenase P (0,2mg/mL) in lOmL of RPMI (Gibco) and incubated for 40minutes at 37°C on GentleMACS (Miltenyi) with the appropriate program. Following dissociation, tissues were filtered (70pm, Clearline), centrifuged 5 min at 250g, and resuspended in PBS.
  • Skeletal muscles from PIK3CA WT and PIK3CA HSA ⁇ CreER mice were isolated, rinsed in phosphate buffered saline IX, cute in small pieces and digested as detailed below. After dissociation, cell suspensions were filtered (70pm, Clearline), centrifuged 10 min at 350g, and resuspended in phosphate buffered saline solution IX supplemented with 2% fetal bovine serum and EDTA at 0.5 m/mol. Cells were then transferred in microtube. Samples were run on an ImageStream ISX mkll (Amnis part of Luminex) that combines flow cytometry with detailed cell imaging. Magnification (40X) was used for all acquisitions. Data were acquired with INSPIRE software (Amnis part of Luminex) and analyzed with IDEAS software (v.6.2, Amnis part of Luminex).
  • Plasma and serum were obtained after centrifugation of the blood at 500 g for 10 min. Blood samples were obtained in EDTA tubes for plasma analysis and EDTA-free tubes for serum analysis. Cells, plasma and serum samples were immediately snap-frozen in liquid nitrogen. For the LC-MS analyses, metabolites were extracted as previously described 9 . Briefly, the extraction solution was composed of 50% methanol, 30% ACN, and 20% water. The volume of the added extraction solution was adjusted to the cell number (1 ml per 1 million cells) or plasma and serum volume (200 pl per 10 pl of plasma or serum). After the addition of extraction solution, samples were vortexed for 5 min at 4 C and then centrifuged at 16,000 g for 15 min at 4 C.
  • LC-MS analyses were conducted using a QExactive Plus Orbitrap mass spectrometer (Thermo) equipped with an Ion Max source and a HESI II probe coupled to a Dionex UltiMate 3000 UPLC system (Thermo). External mass calibration was performed using the standard calibration mixture every 7 d as recommended by the manufacturer. Five microliters of each sample was injected onto Zic-pHilic (150 mm * 2.1 mm i.d. 5 pm) with the guard column (20 mm * 2.1 mm i.d. 5 pm) (Millipore) for liquid chromatography separation.
  • Buffer A was 20 mM ammonium carbonate and 0.1% ammonium hydroxide (pH 9.2); buffer B was acetonitrile.
  • the chromatographic gradient was run at a flow rate of 0.200 pl/min as follows: for 0-20 min, linear gradient from 80 to 20% B; for 20-20.5 min, linear gradient from 20 to 80% B; for 20.5-28 min, hold at 80% B.
  • the mass spectrometer was operated in full-scan polarity switching mode with the spray voltage set to 2.5 kV and the heated capillary held at 320 C.
  • the sheath gas flow was set to 20 units, the auxiliary gas flow was set to 5 units, and the sweep gas flow was set to 0 units.
  • the metabolites were detected across a mass range of 75- 1000 m/z at a resolution of 35,000 (at 200 m/z) with the AGC target set to 106 and a maximum injection time of 250 ms. Lock masses were used to ensure mass accuracy below 5 ppm. Data were acquired with Thermo Xcalibur 4.0.27.131 software (Thermo). The peak areas of metabolites were determined using Thermo TraceFinder 3.3 SP1 software (Thermo) and identified by the exact mass of each singly charged ion and by known retention time in the HPLC column. Patients
  • HFMH congenital facial asymmetry with thickening of soft tissues leading to a set of specific clinical features: narrow palpebral fissure, eyebrow ptosis, nose deviation with small alar rim, external ear asymmetry and displacement with prominent concha, chin deviation with skin dimpling, and lip commissure canting.
  • Alpelisib was compassionately offered by Novartis. Biopsies of the buccinator and/or masseter muscles of the affected and control sides were performed in 5/5 HFMH patients for diagnosis. Control buccinator and masseter muscles were harvested on pediatric patients admitted for facial lacerations and/or mandibular trauma, that required surgical approaches to this anatomical region.
  • EMG needle electromyogram
  • mice In order to gain insight into the mechanisms of PIK3CA -related muscle overgrowth we designed a mouse model carrying specifically a PIK3CA gain-of-function mutation in striated muscle. To this aim, we interbred the R26StopFLPl 10* mouse strain with HSA Cre mice to generate PIK3CA HSA ⁇ CreER animals that express a constitutively overactivated form of PIK3CA upon tamoxifen administration. To follow Cre recombination, PIK3CA HSA ⁇ CreER mice were then interbred with Gt(ROSA)26Sortm4(ACTB-tdTomato-EGFP)Luo/J mice.
  • mice express a cell membrane-localized tdTomato fluorescent protein that is replaced by GFP after Cre recombination.
  • PIK3CA HSA ⁇ CreER mice progressively gained weight compared to their wild-type littermates (PIK3CA WT ) ( Figure 1A). This was the case for both males and females. Around 11 weeks post induction in males and 24 weeks in females, body weight of controls mice became higher than in PIK3CA HSA ⁇ CreER ( Figure 1A).
  • PI3K have been involved in mitochondrial function 16 , we therefore explored the impact of PIK3CA gain-of-function on mitochondria number and activity in the PIK3CA HSA ⁇ CreER mouse model.
  • Flow cytometry of isolated striated muscle cells revealed alteration in the mitochondrial transmembrane potential as assessed by tetramethylrhodamine ethyl ester (TMRE) staining with reduced mitotracker expression (Figure IP and IQ).
  • a PIK3CA inhibitor prevents and reduces striated muscle hypertrophy in a PIK3CA HSA ⁇ CreER mouse model
  • VAF Variant allele frequency
  • COSMIC Catalogue Of Somatic Mutations In Cancer

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Neurology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Zoology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Les inventeurs ont supposé que la voie PIK3CA/AKT/mTOR était anormalement affectée chez des patients atteints de HFMH. Ils ont identifié une mutation gain de fonction somatique de PIK3CA chez 5 patients pédiatriques ayant une myohyperplasie hémifaciale, une cause rare d'asymétrie faciale impliquant exclusivement des muscles faciaux. Pour comprendre la physiopathologie de l'hypertrophie musculaire, ils ont créé un modèle de souris portant spécifiquement une mutation PIK3CA dans les muscles squelettiques. La mutation gain de fonction PIK3CA a conduit à une hypertrophie des cellules musculaires striées, un dysfonctionnement des mitochondries et une hypoglycémie à faibles taux d'insuline circulants. Un inhibiteur de PIK3CA, à savoir l'Alpelisib, a été capable d'empêcher et de réduire l'hypertrophie musculaire dans le modèle de souris. Ils concluent ensuite que PIK3CA est une cible pertinente pour induire une hypertrophie musculaire squelettique. La présente invention concerne une protéine PIK3CA ou un fragment de celle-ci et/ou un agent pour l'expression de la protéine PIK3CA destiné à être utilisé pour favoriser l'hypertrophie musculaire squelettique chez un sujet en ayant besoin.
PCT/EP2024/074017 2023-08-29 2024-08-28 Procédés pour induire une hypertrophie musculaire Pending WO2025045915A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP23306430 2023-08-29
EP23306430.2 2023-08-29

Publications (1)

Publication Number Publication Date
WO2025045915A1 true WO2025045915A1 (fr) 2025-03-06

Family

ID=88017685

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2024/074017 Pending WO2025045915A1 (fr) 2023-08-29 2024-08-28 Procédés pour induire une hypertrophie musculaire

Country Status (1)

Country Link
WO (1) WO2025045915A1 (fr)

Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4179337A (en) 1973-07-20 1979-12-18 Davis Frank F Non-immunogenic polypeptides
US4861719A (en) 1986-04-25 1989-08-29 Fred Hutchinson Cancer Research Center DNA constructs for retrovirus packaging cell lines
WO1990013540A1 (fr) 1989-04-19 1990-11-15 Enzon, Inc. Carbonates actifs d'oxydes de polyalkylene pour la modification de polypeptides
US5278056A (en) 1988-02-05 1994-01-11 The Trustees Of Columbia University In The City Of New York Retroviral packaging cell lines and process of using same
WO1994019478A1 (fr) 1993-02-22 1994-09-01 The Rockefeller University Production de retrovirus exempts d'auxiliaires, a titre eleve par transfection transitoire
WO1995014785A1 (fr) 1993-11-23 1995-06-01 Rhone-Poulenc Rorer S.A. Composition pour la production de produits therapeutiques in vivo
WO1996022378A1 (fr) 1995-01-20 1996-07-25 Rhone-Poulenc Rorer S.A. Cellules pour la production d'adenovirus recombinants
US5882877A (en) 1992-12-03 1999-03-16 Genzyme Corporation Adenoviral vectors for gene therapy containing deletions in the adenoviral genome
US6013516A (en) 1995-10-06 2000-01-11 The Salk Institute For Biological Studies Vector and method of use for nucleic acid delivery to non-dividing cells
WO2001037852A2 (fr) * 1999-11-23 2001-05-31 Regeneron Pharmaceuticals, Inc. Methodes permettant d'inhiber l'atrophie ou de favoriser l'hypertrophie
US20020123456A1 (en) * 2001-03-02 2002-09-05 Glass David J. Methods of identifying agents affecting atrophy and hypertrophy
ES2322881A1 (es) * 2005-12-26 2009-06-30 Consejo Superior Investig. Cientificas Uso de compuestos agonistas de la actividad pi3k en la elaboracion de composiciones farmaceuticas, dichas composiciones farmaceuticas y sus aplicaciones en el tratamiento de enfermedades neurodegenerativas.
ES2547256T3 (es) * 2007-05-18 2015-10-02 Glaxosmithkline Llc Derivados de quinolina como inhibidores de la PI3 cinasa
US20160287601A1 (en) * 2013-10-03 2016-10-06 Millennium Pharmaceuticals, Inc. Enhanced treatment regimens using pi3k inhibitors
KR20190108777A (ko) * 2018-03-15 2019-09-25 숙명여자대학교산학협력단 진세노사이드 Rb1 및 Rb2를 유효성분으로 함유하는 근육 재생용 조성물
US20220323428A1 (en) * 2016-02-19 2022-10-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Byl719 (alpelisib) for use in the treatment of pik3ca-related overgrowth spectrum (pros-cloves syndrome)
WO2023041905A1 (fr) 2021-09-14 2023-03-23 Ucl Business Ltd Aminopyridines servant d'activateurs de la pi3-kinase

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4179337A (en) 1973-07-20 1979-12-18 Davis Frank F Non-immunogenic polypeptides
US4861719A (en) 1986-04-25 1989-08-29 Fred Hutchinson Cancer Research Center DNA constructs for retrovirus packaging cell lines
US5278056A (en) 1988-02-05 1994-01-11 The Trustees Of Columbia University In The City Of New York Retroviral packaging cell lines and process of using same
WO1990013540A1 (fr) 1989-04-19 1990-11-15 Enzon, Inc. Carbonates actifs d'oxydes de polyalkylene pour la modification de polypeptides
US5882877A (en) 1992-12-03 1999-03-16 Genzyme Corporation Adenoviral vectors for gene therapy containing deletions in the adenoviral genome
WO1994019478A1 (fr) 1993-02-22 1994-09-01 The Rockefeller University Production de retrovirus exempts d'auxiliaires, a titre eleve par transfection transitoire
WO1995014785A1 (fr) 1993-11-23 1995-06-01 Rhone-Poulenc Rorer S.A. Composition pour la production de produits therapeutiques in vivo
WO1996022378A1 (fr) 1995-01-20 1996-07-25 Rhone-Poulenc Rorer S.A. Cellules pour la production d'adenovirus recombinants
US6013516A (en) 1995-10-06 2000-01-11 The Salk Institute For Biological Studies Vector and method of use for nucleic acid delivery to non-dividing cells
WO2001037852A2 (fr) * 1999-11-23 2001-05-31 Regeneron Pharmaceuticals, Inc. Methodes permettant d'inhiber l'atrophie ou de favoriser l'hypertrophie
US20020123456A1 (en) * 2001-03-02 2002-09-05 Glass David J. Methods of identifying agents affecting atrophy and hypertrophy
ES2322881A1 (es) * 2005-12-26 2009-06-30 Consejo Superior Investig. Cientificas Uso de compuestos agonistas de la actividad pi3k en la elaboracion de composiciones farmaceuticas, dichas composiciones farmaceuticas y sus aplicaciones en el tratamiento de enfermedades neurodegenerativas.
ES2547256T3 (es) * 2007-05-18 2015-10-02 Glaxosmithkline Llc Derivados de quinolina como inhibidores de la PI3 cinasa
US20160287601A1 (en) * 2013-10-03 2016-10-06 Millennium Pharmaceuticals, Inc. Enhanced treatment regimens using pi3k inhibitors
US20220323428A1 (en) * 2016-02-19 2022-10-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Byl719 (alpelisib) for use in the treatment of pik3ca-related overgrowth spectrum (pros-cloves syndrome)
KR20190108777A (ko) * 2018-03-15 2019-09-25 숙명여자대학교산학협력단 진세노사이드 Rb1 및 Rb2를 유효성분으로 함유하는 근육 재생용 조성물
WO2023041905A1 (fr) 2021-09-14 2023-03-23 Ucl Business Ltd Aminopyridines servant d'activateurs de la pi3-kinase

Non-Patent Citations (19)

* Cited by examiner, † Cited by third party
Title
BANKHEAD, P. ET AL.: "QuPath: Open source software for digital pathology image analysis", SCI REP, vol. 7, 2017, pages 16878, XP055452757, Retrieved from the Internet <URL:https://doi.org:10.1038/s41598-017-17204-5> DOI: 10.1038/s41598-017-17204-5
CANAUD, G.HAMMILL, A. M.ADAMS, D.VIKKULA, M.KEPPLER-NOREUIL, K. M.: "A review of mechanisms of disease across PIK3CA-related disorders with vascular manifestations", ORPHANET J RARE DIS, vol. 16, 2021, pages 306, Retrieved from the Internet <URL:https://doi.org:10.1186/s13023-021-01929-8>
CASTILLO TAUCHER, S.PARDO VARGAS, R. A.CAIVI LEON, D.: "Hemifacial myohyperplasia: an additional case", AM J MED GENET A, vol. 116A, 2003, pages 103 - 104
DELESTRE, F. ET AL.: "Alpelisib administration reduced lymphatic malformations in a mouse model and in patients", SCIENCE TRANSLATIONAL MEDICINE, vol. 13, 2021, pages eabg0809, Retrieved from the Internet <URL:https://doi.org:10.1126/scitranslmed.abg0809>
DOYON G ET AL.: "Discovery of a Small Molecule Agonist of Phosphatidylinositol 3-Kinase pl 10a That Reactivates Latent HIV-1", PLOS ONE, vol. 9, no. 1, 2014, pages e84964
GLASS DAVID J: "Molecular mechanisms modulating muscle mass", TRENDS IN MOLECULAR MEDICINE, ELSEVIER CURRENT TRENDS, GB, vol. 9, no. 8, 1 August 2003 (2003-08-01), pages 344 - 350, XP002332242, ISSN: 1471-4914, DOI: 10.1016/S1471-4914(03)00138-2 *
GRACE Q ET AL.: "A small-molecule PI3Ka activator for cardioprotection and neuroregeneration.", NATURE, vol. 618, no. 7963, 2023, pages 159 - 168
KLIPPEL, A. ET AL.: "Membrane localization of phosphatidylinositol 3-kinase is sufficient to activate multiple signal-transducing kinase pathways", MOLECULAR AND CELLULAR BIOLOGY, vol. 16, 1996, pages 4117 - 4127, XP002037799
LADRAA, S. ET AL.: "PIK3CA gain-of-function mutation in adipose tissue induces metabolic reprogramming with Warburg-like effect and severe endocrine disruption", SCI ADV, vol. 8, 2022, pages eade7823, Retrieved from the Internet <URL:https://doi.org:10.1126/sciadv.ade7823>
LEE, S.SZE, R.MURAKAMI, C.GRUSS, J.CUNNINGHAM, M.: "Hemifacial myohyperplasia: description of a new syndrome", AMERICAN JOURNAL OF MEDICAL GENETICS, vol. 103, 2001, pages 326 - 333
LI, M. E. ET AL.: "Role of p110a subunit of PI3-kinase in skeletal muscle mitochondrial homeostasis and metabolism", NAT COMMUN, vol. 10, 2019, pages 3412, Retrieved from the Internet <URL:https://doi.org:10.1038/s41467-019-11265-y>
MADSEN, R. R.VANHAESEBROECK, B.: "Cracking the context-specific PI3K signaling code", SCIENCE SIGNALING, vol. 13, 2020, Retrieved from the Internet <URL:https://doi.org:10.1126/scisignal.aay2940>
MIRANDA, R. T.BARROS, L. M.SANTOS, L. A.BONAN, P. R.MARTELLI, H., JR: "Clinical and imaging features in a patient with hemifacial hyperplasia", J ORAL SCI, vol. 52, 2010, pages 509 - 512
PEREIRA-PERDOMO, D. F.VELEZ-FORERO, J.PRADA-MADRID, R.: "Hemifacial myohyperplasia sequence", AM J MED GENET A, vol. 152A, 2010, pages 1770 - 1773, XP072324474, Retrieved from the Internet <URL:https://doi.org:10.1002/ajmg.a.33428> DOI: 10.1002/ajmg.a.33428
SCHINDELIN, J. ET AL.: "Fiji: an open-source platform for biological-image analysis", NATURE METHODS, vol. 9, 2012, pages 676 - 682, XP055343835, Retrieved from the Internet <URL:https://doi.org:10.1038/nmeth.2019> DOI: 10.1038/nmeth.2019
SIPONEN, M.SANDOR, G. K.YLIKONTIOLA, L.SALO, T.TUOMINEN, H.: "Multiple orofacial intraneural perineuriomas in a patient with hemifacial hyperplasia", ORAL SURG ORAL MED ORAL PATHOL ORAL RADIOL ENDOD, vol. 104, 2007, pages e38 - 44, XP022129656, Retrieved from the Internet <URL:https://doi.org:10.1016/j.tripleo.2006.12.030> DOI: 10.1016/j.tripleo.2006.12.030
SRINIVASAN, L. ET AL.: "PI3 kinase signals BCR-dependent mature B cell survival", CELL, vol. 139, 2009, pages 573 - 586, Retrieved from the Internet <URL:https:Hdoi.org:10.1016/j.cell.2009.08.041>
STRINGER, C.WANG, T.MICHAELOS, M.PACHITARIU, M.: "Cellpose: a generalist algorithm for cellular segmentation", NATURE METHODS, vol. 18, 2021, pages 100 - 106, XP037330201, Retrieved from the Internet <URL:https://doi.org:10.1038/s41592-020-01018-x> DOI: 10.1038/s41592-020-01018-x
ZISSMAN, S.COOPERMAN, Y.LESHEM, D.GUR, E.: "Progressive Surgical Management of Hemifacial Myohyperplasia for Improved Functional and Aesthetic Results", PLAST RECONSTR SURG GLOB OPEN, vol. 8, 2020, pages e2724, Retrieved from the Internet <URL:https://doi.org:10.1097/GOX.0000000000002724>

Similar Documents

Publication Publication Date Title
JP6634402B2 (ja) 代謝障害及び疾患の治療のための組成物、使用及び方法
JP6936366B2 (ja) 胆汁酸に関係した障害の治療方法
CN104147591B (zh) Jnk信号转导通路的细胞可渗透性肽抑制剂用于治疗各种疾病的应用
RU2729161C2 (ru) Фармацевтические композиции, содержащие варианты пептидов, и способы их применения
JP5290966B2 (ja) 安定化されたインスリン様増殖因子ポリペプチド
AU2008257448B9 (en) Unacylated ghrelin as therapeutic agent in the treatment of metabolic disorders
KR102508651B1 (ko) 신경계 및 신경변성 상태의 요법으로서 장기간 작용하는 glf-1r 작용제
JP6576251B2 (ja) ＰＴＤ−Ｓｍａｄ７薬物療法
KR20200024263A (ko) 고혈당증의 치료 및 예방을 위한 펩타이드
KR102819396B1 (ko) 호모시스틴뇨증의 치료를 위한 조성물 및 방법
US20220287347A1 (en) Peptides for treating muscle atrophy
CN105792839A (zh) 用于治疗的胰岛素样生长因子拟似物
WO2025045915A1 (fr) Procédés pour induire une hypertrophie musculaire
US20200030413A1 (en) Methods and pharmaceutical compositions for the treatment of kidney cancer
KR20150140686A (ko) 신증후군 및 관련 병태의 치료 방법
EP3609577B1 (fr) Adipokines sélectives vis-à-vis de la graisse brune
EP4021480A1 (fr) Peptides pour le traitement de troubles rénaux
AU2017274094B2 (en) Compositions and methods for treating metabolic diseases
WO2023010049A1 (fr) Utilisations thérapeutiques de la protéine isthmine
US20240277816A1 (en) Polypeptide conjugate and application thereof in preparation of drug for treating diseases related to glucose metabolism
CN104470532A (zh) SorCS1在治疗肥胖症及超重中的应用
EP4545570A1 (fr) Protéine de fusion contenant un agoniste du récepteur glp-1 amélioré et utilisations
US20250145994A1 (en) Compositions and methods for treating conditions related to cellular senescence
US20180369325A1 (en) Compounds and methods for treating metabolic disorders
WO2025215206A1 (fr) Activateur de cils primaires pour traiter des dysfonctionnements cognitifs ou des ciliopathies

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24762648

Country of ref document: EP

Kind code of ref document: A1