WO2016161086A1 - Méthodes et compositions pour le traitement de glycogénoses à l'aide d'agents imitant l'amp cyclique ou élevant celui-ci - Google Patents

Méthodes et compositions pour le traitement de glycogénoses à l'aide d'agents imitant l'amp cyclique ou élevant celui-ci Download PDF

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WO2016161086A1
WO2016161086A1 PCT/US2016/025215 US2016025215W WO2016161086A1 WO 2016161086 A1 WO2016161086 A1 WO 2016161086A1 US 2016025215 W US2016025215 W US 2016025215W WO 2016161086 A1 WO2016161086 A1 WO 2016161086A1
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glycogen
gsd
available
camp
therapy
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Baodong Sun
Priya Kishnani
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Duke University
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    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
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    • C12Y207/11013—Protein kinase C (2.7.11.13)

Definitions

  • Glycogen is a branched polymer of glucose that serves as a form of energy storage in human and animals.
  • the two major sites of glycogen storage are liver and muscle.
  • the primary function of glycogen varies in different tissues.
  • glycogen serves as a glucose reserve for the maintenance of blood-glucose levels; in muscle, glycogen provides energy for muscle contraction [1,2].
  • Glycogen metabolism is a complex process involving many different enzymes that directly or indirectly regulate glycogen synthesis and degradation.
  • glycogen is synthesized in the cytosol by the two enzymes, glycogen synthase (GS) and glycogen branching enzyme (GBE).
  • glycogen is degraded in the cytoplasm (glycogenolysis) by a combined action of glycogen phosphorylase (GP) (cleaves the ⁇ -1,4-glycosidic bonds) and glycogen debranching enzyme (GDE) (cleaves ⁇ -1,6-glycosidic bonds at the branch points), but a very small amount of glycogen is transported into lysosomes and digested into glucose by the enzyme acid alpha-glucosidase (GAA) [2,3].
  • GP glycogen phosphorylase
  • GDE glycogen debranching enzyme
  • Glycogen synthesis and degradation involve numerous enzymes. Mutations in genes encoding these enzymes cause a partial or complete loss of the enzyme activities in glycogen storage diseases (GSDs), a group of genetic disorders with abnormal metabolism of glycogen primarily in liver, muscle, and the brain. Most of the GSDs are inherited in an autosomal recessive manner, some are X linked or inherited in an autosomal dominant manner. The overall frequency of all forms of glycogen storage diseases is approximately 1 in 10,000 live births. There are over 13 forms of GSD presently identified, and a wide spectrum of clinical presentations is seen. GSD types I, II, III, VI, and IX are currently recognized as the most common forms, accounting for over 90 percent of all cases.
  • GSD II a deficiency in the lysosomal enzyme acid alpha-glucosidase (GAA)
  • GAA acid alpha-glucosidase
  • Enzyme replacement therapy would be an ideal treatment for genetic diseases with singly-gene deficiency.
  • ERT has been effective in diseases in which the responsible enzymes/proteins exert their functions in extracellular fluids, such as adenosine deaminase deficiency, hemophilia, and alpha l-antitrypsin deficiency, or in a lysosomal location such as lysosomal storage diseases including Pompe disease in which the therapeutic enzyme could be efficiently delivered into the lysosome of diseased cells via a mannose-6-phosphate receptor (M6PR or IGF2)-mediated uptake.
  • M6PR mannose-6-phosphate receptor
  • ERT with recombinant human GAA is the only approved therapy for Pompe disease.
  • ERT has been successful only in some patients as challenges with low targeting efficiency in skeletal muscle and immunogenicity among others. There is therefore an urgent need to develop new therapies for the treatment of other GSDs especially a therapy that is suitable for most, if not all, GSDs.
  • compositions for treating glycogen storage diseases are provided herein.
  • administration of cyclic AMP elevators is shown to reduce glycogen storage in affected cells and thus can be used to treat or reduce symptoms in subjects with glycogen storage diseases or other conditions where there is a build of glycogen.
  • the methods of treating a glycogen storage disease or other conditions where there is a build of glycogen in a subject include administering a therapeutically effective amount of a cyclic AMP elevator to the subject in need of treatment.
  • the methods and compositions can further include administering an enzyme replacement therapy, a gene replacement therapy, a chaperone therapy, or a substrate reduction therapy using RNAi-based treatment approaches, antisense therapies, or small molecule or peptide drugs to the subject.
  • One embodiment of the invention provides a method of treating a glycogen storage disease or other conditions where there is a build of glycogen in a subject in need of such treatment comprising administering a therapeutically effective amount of a cyclic AMP elevator to the subject in need of treatment for a glycogen storage disease, or a condition where there is a buildup of glycogen from other disorders.
  • the glycogen storage disease or other condition where there is a build of glycogen can be selected from the group consisting of GSD I, GSD II (Pompe disease), GSD III, GSD IV, GSD V, GSD VI, GSD VII, GSD IX, GSD XI, GSD XII, GSD XIII, GSD XIV, Danon disease, Fanconi-Bickel disease, Lafora disease, cardiac/muscle glycogenosis due to AMP-activated protein kinase gamma subunit 2- deficiency (PRKAG2 cardiac syndrome), or other disorders where there is secondary accumulation of glycogen, such as Niemann-Pick Disease, GSD X, phosphoglycerate kinase deficiency, RBCK1 deficiency, and GSD XV.
  • the cyclic AMP elevator can be an adenylate cyclase activator, a phosphodiesterase (PDE) inhibitor, a Toll-like receptor ligand, a calcium ionophore, a protein kinase A (PKA) activator, a protein kinase C (PKC) activator, a beta2-adrenergic receptor agonist, an adenylate cyclase toxin, or combinations thereof.
  • PDE phosphodiesterase
  • Toll-like receptor ligand a calcium ionophore
  • PKA protein kinase A
  • PDC protein kinase C
  • beta2-adrenergic receptor agonist an adenylate cyclase toxin, or combinations thereof.
  • the adenylate cyclase activator can be a labdane diterpene, a G- protein coupled receptor agonist, a G-protein activator, the pyrazole derivative A02011-1, benzyloxybenzaldehyde and analogs thereof, or combinations thereof.
  • the labdane diterpene can be labdane, forskolin, a forskolin derivative, 6-acetyl-7-deacetyl-forskolin, 7-deacetyl-forskolin, 7-deacetyl-6-(N- acetylglycyl)-forskolin, 7-deacetyl-7-O-hemisuccunyl-forskolin, 7-deacetyl-7- (O-N-methylpiperazino)- ⁇ -butryl-dihydrochlonde-forskolin, 7-HPP-forskolin, 6- HPP-forskolin, colforsin daropate hydrochloride (NKH477), or combinations thereof.
  • the G-protein coupled receptor agonist can be catecholamine, dopamine, dobutamine, isoproterenol, adenosine, carbacyclin, endothelin, epinephrine, glucagon, octopamine, pituitary adenylate cyclase-activating peptide (PACAP), parathyroid hormone, prostaglandin, vasopressin, or combinations thereof.
  • PACAP pituitary adenylate cyclase-activating peptide
  • the G-protein activator can be cholera toxin or a subunit thereof.
  • the PDE inhibitor can be a PDE3, PDE4, PDE7, or PDE8 inhibitor.
  • the Toll-like receptor ligand can be lipopolysaccharide (LPS), 1- palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (pLPC), lipoteichoic acid (LTA), flagellin, or combinations thereof.
  • the calcium ionophore can be an ionomycin calcium salt, A23187, or combinations thereof.
  • the PKA activator can be 6-Bnz-cAMP, 8-CPT-2’-O-Me-cAMP, 8-CPT- cAMP, 8-Bromo-cAMP, Dibutyryl-cAMP, Dioctanoyl-cAMP, Sp-8-Br-cAMPS, Sp-cAMPS, cAMP, a PKA subunit, or combinations thereof.
  • the PKC activator can be phorbol myristate acetate (PMA), a PKC purified enzyme, or combinations thereof.
  • PMA phorbol myristate acetate
  • PKC purified enzyme or combinations thereof.
  • the beta2-adrenergic receptor agonist can be bitolterol, fenoterol, isoprenaline, levosalbutamol, orciprenaline, pirbuterol, procaterol, ritodrine, salbutamol, terbutaline, arformoterol, bambuterol, clenbuterol, formoterol, salmeterol, indacaterol, olodaterol, vilanterol, vilanterol with umeclidinium bromide, vilanterol with fluticasone furoate, zilpaterol, or combinations thereof.
  • the methods of the invention can further comprise administering a therapeutically effective amount of an enzyme replacement therapy, a gene therapy, a substrate reduction therapy, a chaperone therapy, or combinations thereof to the subject with a glycogen storage disease (primary or a secondary accumulation of glycogen due to other primary diseases).
  • a glycogen storage disease primary or a secondary accumulation of glycogen due to other primary diseases.
  • Two or more cyclic AMP elevators can administered to the subject.
  • the cyclic AMP elevator can be administered by a route selected from oral, parenteral, intramuscular, intravenous, intraperitoneal and subcutaneous.
  • Another embodiment of the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising at least one cyclic AMP elevator and an enzyme replacement therapy, gene therapy, substrate reduction therapy, chaperone therapy specific for a glycogen storage disease, or combinations thereof.
  • the enzyme replacement therapy or gene therapy can be selected from the group consisting of administration of the following therapeutic enzymes or genes encoding these enzymes: acid alpha-glucosidase, glucose- 6-phosphatase, glycogen debranching enzyme, glycogen branching enzyme, muscle glycogen phosphorylase, liver glycogen phosphorylase, muscle phosphofructokinase, phosphorylase kinase, lactate dehydrogenase, glucose transporter 2, Aldolase A, phosphoglucomutase, Laforin or Malin, LAMP-2, and ⁇ -enolase.
  • the substrate reduction therapy can be selected from the group consisting of methods of inhibiting glycogen synthase to reduce glycogen accumulation, siRNA-based therapies, shRNA-based therapies, antisense therapies, and therapies using small molecules or peptide drugs.
  • kits comprising at least one cyclic AMP elevator, an enzyme replacement therapy or gene therapy and instructions for administering the cyclic AMP elevator and the enzyme replacement therapy or gene therapy to a subject with a glycogen storage disease or other conditions where there is a build of glycogen, wherein the enzyme replacement therapy or gene therapy replaces an enzyme or gene encoding the enzyme deficient in the glycogen storage disease.
  • the kit can further comprise one or more chaperone therapies.
  • kits comprising at least one cyclic AMP elevator, a substrate reduction therapy, and instructions for administering the cyclic AMP elevator and the substrate reduction therapy to a subject with a glycogen storage disease or other condition where there is a build of glycogen, wherein the substrate reduction therapy reduces glycogen synthase activity and prevents glycogen accumulation in the glycogen storage disease.
  • Figure 1 is a schematic showing the role of cyclic AMP in stimulating glycogen breakdown and inhibiting glycogen synthesis through activation of cAMP-dependent protein kinase A (PKA).
  • PKA cAMP-dependent protein kinase A
  • Inactive forms are phosphorylase b, phosphorylase kinase b, and glycogen synthase b.
  • Active forms are phosphorylase kinase a, phosphorylase a, and glycogen synthase a. Adapted from Berridge, M.J., 2014.
  • Figure 2 is a graph showing that Forskolin reduced glycogen content in primary muscle cells from patients with GSD IIIa.
  • UT untreated; Low, 1 ⁇ M Forskolin; High, 10 ⁇ M Forskolin. *, p ⁇ 0.05 vs. UT; **, p ⁇ 0.001 vs. UT.
  • Figure 3 is a graph showing that glycogen content was decreased in GSD IV patient fibroblasts by Forskolin treatment.
  • UT untreated; Low, 1 ⁇ M Forskolin; High, 10 ⁇ M Forskolin. *, p ⁇ 0.05 vs. UT; **, p ⁇ 0.001 vs. UT.
  • This invention relates to methods and compositions of treating or preventing glycogen storage diseases (GSD) type III (Forbes-Cori disease, Debranching enzyme deficiency) and IV (Andersen disease, Branching Enzyme Deficiency), by administering one or more cAMP elevators or agents that mimic cAMP, for example Forskolin or its derivatives or analogs thereof in a therapeutically effective amount.
  • GSD glycogen storage diseases
  • IV Andersen disease, Branching Enzyme Deficiency
  • the invention also relates to use of the same method as an adjunctive therapy in combination with other therapies, such as enzyme replacement therapy, gene therapy, substrate reduction, chaperone therapy, and other therapeutic approaches.
  • the invention further relates to use of the same method, either alone or as an adjunctive therapy, for treating other GSDs and other disorders where there is a buildup of glycogen including, but not limited to, type I (von Gierke disease, glucose-6- phosphatase deficiency), II (Pompe disease, acid-alpha-glucosidase deficiency), V (McArdle disease, Muscle Phosphorylase Deficiency), VI (Hers disease, Liver phosphorylase enzyme), VII (Tarui disease, Muscle Phosphofructokinase Deficiency), IX (Phosphorylase Kinase Deficiency), XI (Lactate dehydrogenase deficiency), XII (Aldolase A deficiency), XIII ( ⁇ - enolase deficiency), GSD XIV (Phosphoglucomutase deficiency); Fanconi- Bickel disease (deficiency in glucose
  • Glycogen synthesis and degradation are reciprocally regulated by hormonal signals. Insulin and glucagon (or epinephrine) are the major hormones that regulate glycogen storage and mobilization. Glycogen synthesis is triggered when blood-glucose levels are high, through insulin- induced activation of protein phosphatase-1 (PP1). PP1 activates GS (b ⁇ a) and inactivates GP and glycogen phosphorylase kinase (GPK) (a ⁇ b) by dephosphorylating these enzymes ( Figure 1).
  • PP1 protein phosphatase-1
  • GPK glycogen phosphorylase kinase
  • glycogen breakdown is initiated in a starved state by glucagon (in liver) or epinephrine (in muscle) triggered cyclic AMP (cAMP) cascade acting through cAMP-dependent protein kinase A (PKA) ( Figure 1).
  • glucagon in liver
  • epinephrine in muscle
  • cAMP cyclic AMP
  • PKA stimulates glycogen degradation by the following actions: 1) phosphorylates and activates GPK (b ⁇ a), which subsequently phosphorylates and activates GP (b ⁇ a); 2) phosphorylates GS (a ⁇ b), which leads to a decrease in enzymatic activity to prevent glycogen being synthesized at the same time that it is being broken down; and 3) disables PP1 function by dissociating it from glycogen through phosphorylation of the glycogen-binding subunit G M ( Figure 1) [3,7,8,9].
  • GSD III Glycogen debranching enzyme deficiency, Cori Disease
  • GDE glycogen debranching enzyme
  • the progressive myopathy and/or cardiomyopathy and/or peripheral neuropathy are major causes of morbidity in adults [10,11,12].
  • Current treatment is symptomatic, and there is no effective therapy for the disease.
  • Hypoglycemia can be controlled by frequent meals high in carbohydrates with cornstarch supplements or nocturnal gastric drip feedings.
  • Patients with myopathy have been treated with a diet high in protein during the daytime plus overnight enteral infusion. In some patients transient improvement in symptoms has been documented, but there are no systemic studies or long-term data demonstrating that the high protein diet prevents or treats the progressive myopathy [10]. These approaches do little to alter the long term course and morbidity of these diseases.
  • Curly coated retrievers with a phenotype mimicking type IIIa disease have been identified. These dogs have hepatomegaly, hypoglycemia, and elevated liver enzymes and creatine phosphokinase. The clinical signs in these affected dogs appear to be mild in the first year of life, becoming more prominent with age and leading to lethargy, exercise intolerance, and episodic hypoglycemia with collapse/unresponsiveness. These dogs are homozygous for the c.4223delA mutation [13]. Rapamycin, a specific inhibitor of mTOR, can significantly reduce glycogen content in both liver and skeletal muscle of affected CCR dogs [14]. This suggests suppression of glycogen synthesis with Rapamycin is a potential useful therapy for GSD III.
  • GSD IV Glycogen branching enzyme deficiency; Andersen Disease
  • GSD IV is a rare autosomal recessive disorder caused by deficiency of glycogen branching enzyme (GBE), a key enzyme involved in glycogen synthesis.
  • GSD IV is clinically variable.
  • the classical form of GSD IV is characterized by failure to thrive, hepatosplenomegaly, and progressive liver cirrhosis which normally lead to death by age 5 years. Some patients can develop hepatic adenomas and hepatocellular carcinoma.
  • the perinatal form is characterized by multiple congenital contractures, hydrops fetalis, and perinatal death.
  • the congenital form includes hypotonia, muscle wasting, neuronal involvement, inconsistent cardiomyopathy, and death in early infancy.
  • patients present predominantly with a myopathy or cardiomyopathy.
  • the adult form can present as an isolated myopathy or as a multisystem disorder with central and peripheral nervous system dysfunction accompanied by accumulation of polyglucosan material in the nervous system (so-called adult polyglucosan body disease) [18,19,20].
  • GSD IV Maintenance of normoglycemia and adequate nutrient intake improve liver function and muscle strength in some patients.
  • liver transplantation is the only treatment option [4].
  • Norwegian forest cats are a naturally occurring animal model of GSD IV, caused by an inherited, recessive mutation in the branching enzyme gene [21]. Most homozygous affected kittens die at or soon after birth due to hypoglycemia. The surviving cats appear clinically normal until 5 month of age, when skeletal muscle, heart, and CNS degeneration become obvious, accompanied by elevated body temperature, but cirrhosis and liver failure are absent.
  • a mouse model of GSD IV is available.
  • the homozygous mice (Gbe1(neo/neo)) exhibit a phenotype similar to juvenile onset GSD IV, with wide spread accumulation of polyglucosan bodies [22]. Other mice models of GSDs are also available.
  • GSDs cytoplasmic GSDs
  • GSD I von Gierke’s disease, glucose-6-phosphatase deficiency, Ib translocase deficiency
  • GSD V McArdle’s disease, a deficiency in muscle phosphorylase
  • GSD VI Hers' disease, a deficiency in liver phosphorylase
  • GSD VII a deficiency in muscle phosphofructokinase; Tarui’s disease
  • GSD IX phosphorylase kinase deficiency
  • GSD XI Lactate dehydrogenase deficiency
  • GSD XII Aldolase A deficiency
  • GSD XIII a deficiency in ⁇ - enolase
  • GSD 0 A deficiency in glycogen synthase
  • Fanconi-Bickel disease deficiency in glucose transporter GLUT2
  • Lafora disease laforin
  • cAMP elevator refers to an agent that increases intracellular levels of cAMP beyond the background physiological intracellular level.
  • Intracellular levels of cAMP can be measured by, for example, assays that measure cAMP levels through protein kinase A (PKA), which is activated upon release of its regulatory subunits after binding to cAMP (e.g., cAMP- GLOTM Max assay).
  • PKA protein kinase A
  • cAMP levels can be increased by about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 percent or more (or any range between about 5 and 100 percent).
  • cAMP is synthesized from ATP by the enzyme adenylate cyclase and is degraded into AMP by cAMP phosphodiesterases.
  • cAMP elevators therefore include agents that activate or enhance the activity of adenylate cyclase (hereinafter referred to as“adenylate cyclase activators”), agents that increase the availability of adenylate cyclase, and agents that inhibit or block the activity of cAMP and/or cGMP phosphodiesterases (hereinafter referred to as“PDE inhibitors”).
  • the activity of adenylate cyclase can be measured by, for example, ELISA.
  • the activity of adenylate cyclase can be increased by about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 percent or more (or any range between about 5 and 100 percent).
  • adenylate cyclase can be measured by, for example, ELISA.
  • the availability of adenylate cyclase can be increased by about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 percent or more (or any range between about 5 and 100 percent).
  • the activity of cAMP phosphodiesterases or cGMP phosphodiesterases can be measured by, for example, a PDELightTM Assay Kit (Lonza, Basel, Switzerland).
  • the activity of cAMP phosphodiesterases or cGMP phosphodiesterases can be decreased by about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 percent or more (or any range between about 5 and 100 percent).
  • cAMP elevators include, but are not limited to, the Toll-like receptor ligands, calcium channel activators or calcium activators, beta2-adrenergic receptor agonists, protein kinase C (PKC) activators, and adenylate cyclase toxin. These classes of cAMP elevators are described in more detail herein below.
  • the cAMP elevator is an adenylate cyclase activator, more particularly, a labdane diterpene such as Forskolin or a derivative or analog thereof.
  • agents that mimic cAMP refers to an agent that produces physiological effects similar to endogenous cAMP such as, for example, activating protein kinase A (PKA) (also known as cAMP-dependent enzyme).
  • PKA protein kinase A
  • agents that mimic cAMP include, for example, PKA activators.
  • the activity of PKA activators can be determined using an ELISA (for example, an ELISA that utilizes a synthetic peptide as a substrate for PKA and a polyclonal antibody that recognizes the phosphorylated form of the substrate).
  • a PKA activator can increase protein kinase A activity by about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 percent or more (or any range between about 5 and 100 percent).
  • Forskolin (also called Coleonol) is a chemical that is extracted from the roots of the Indian Coleus plant (Coleus forskohlii). Forskolin is commonly used as a cAMP elevator to raise levels of cAMP in the research of cell physiology. Forskolin activates the enzyme adenylyl cyclase and increases intracellular levels of cAMP [23,24,25,26].
  • Forskolin could induce glycogenolysis and reduce glycogen levels in both cultured cells and experimental animals [27] [28] [29].
  • the role of cyclic AMP in GSD is unknown and the ability of forskolin or another cyclic AMP elevator to treat GSD has not been suggested and would not be predicted to overcome enzyme deficiencies.
  • the ability of some cyclic AMP elevators to cross the blood brain barrier can also make these compounds useful to treat central or peripheral nervous system aspects of disease that are not amenable to treatment with ERT.
  • Forskolin is already in use as a pharmaceutical to treat a number of other unrelated conditions and diseases.
  • a Forskolin dosage can be about 5, 10, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 or more mg in one day and the dose is divided into twice or thrice a day.
  • Forskolin can be administered by intravenously (IV), intramuscularly (IM), subcutaneously (SC), inhalation, oral dosages, eye drops, or any other suitable methods.
  • glycogen levels in GSD III patient muscle cells and GSD IV patient fibroblast cells can significantly reduce glycogen levels in GSD III patient muscle cells and GSD IV patient fibroblast cells (see Examples; Figure 2 and Figure 3), likely through the inhibition of glycogen synthesis.
  • This treatment approach can be extremely effective for the patients with, for example, GSD V/VI and GSD IX who have residual enzyme activities, but the approach can be effective in patients with various GSDs, including but not limited to GSD type I, II, III, IV, V, VI, VII, IX, XI, XII, XIII, XIV, Lafora disease, Fanconi-Bickel disease, Danon disease, PRKAG2 cardiac syndrome, and conditions where there is a secondary buildup of glycogen, such as Niemann-Pick Disease, GSD X (phosphoglycerate mutase deficiency; increased glycogen in muscle); phosphoglycerate kinase deficiency (increased glycogen in muscle); RBCK1 defic
  • cAMP elevators include, but are not limited to, the following types of therapeutic agents: an adenylate cyclase activator, a PDE inhibitor, a Toll-like receptor ligand, a calcium activator, a protein kinase A activator, a protein kinase C activator, a beta2-adrenergic receptor agonist, or a adenylate cyclase toxin.
  • adenylate Cyclase Activity an adenylate cyclase activator, a PDE inhibitor, a Toll-like receptor ligand, a calcium activator, a protein kinase A activator, a protein kinase C activator, a beta2-adrenergic receptor agonist, or a adenylate cyclase toxin.
  • Adenylate cyclase is an enzyme that synthesizes cAMP from ATP. There are at least nine isoforms of adenylate cyclase, which differ considerably in regulatory properties and are differentially expressed among tissues [47, 48]. Early studies indicated that cyclase activity was regulated primarily by interactions with alpha subunits of heterotrimeric G proteins, which are activated through G protein-coupled receptors. More recently, it has become clear that cyclase activity is regulated by multiple effectors, which include not only the alpha subunits of G s and G i proteins, but also the beta- gamma subunits of G proteins and protein kinase C. Five of the adenylate cyclases known are regulated by calcium [49, 50. All known adenylate cyclases are stimulated by exposure of cells to forskolin.
  • adenylate cyclase activators include, but are not limited to, the labdane diterpenes, such as forskolin or a derivative or analog thereof, pyrazole derivatives, benzyloxybenzaldehyde analog, G-protein coupled receptor agonists, and G-protein activators.
  • the cAMP elevator for use within the methods and compositions of the invention is a labdane diterpene such as labdane, forskolin, or a forskolin derivative or analog.
  • the chemical structure for labdane is depicted below:
  • labdane diterpenes are known to those of skill in the art and can also be used as cyclic AMP elevators within the scope used herein and include those disclosed in U.S. Patent No. 5,268,471 to de Souza, U.S. Patent No.5,789,439 to Hosono, U.S. Patent No.4,517,200 to Kreutner, U.S. Patent No. 5,869,523 to de Souza, U.S. Patent No. 5,350,864 to Seamon, U.S. Patent No. 4,871,764 to Schutske. Pharmaceutically acceptable salts of the labdane diterpenes are also included herein.
  • adenylate cyclase activators for use in the methods and compositions of the invention include, but are not limited to, G-protein coupled receptor agonists and G-protein activators.
  • Adenylate cyclase in mammalian cells is normally activated by the stimulatory regulatory protein G s and guanosine triphosphate (GTP); however, the activation is normally brief because an inhibitory regulatory protein (G i ) hydrolyzes the GTP.
  • G i guanosine triphosphate hydrolyzes the GTP.
  • Cholera toxin and pertussis toxin catalyze the covalent incorporation of ADP-ribose into the G-protein ⁇ -subunit [51-54].
  • the pertussis toxin A subunit catalyzes the ADP-ribosylation of G i at a site that impairs the ability of this heterotrimeric G-protein to interact with receptors, thereby blocking the inhibitory effects of G i on adenylate cyclase. In this manner, the conversion of ATP to cAMP is stimulated.
  • the cholera toxin A subunit catalyzes the attachment of ADP- ribose to G s in a manner that stabilizes the GTP-bound form resulting in persistent activation of adenylate cyclase. Purified subunits of these toxins (e.g., cholera toxin A subunit) have also been shown to activate adenylate cyclase.
  • suitable G-protein coupled receptor agonists for use in the methods and compositions of the invention include, but are not limited to, a catecholamine, dopamine, dobutamine, isoproterenol, adenosine, carbacyclin, endothelin, epinephrine, glucagon, octopamine, pituitary adenylate cyclase- activating peptide (PACAP), parathyroid hormone, prostaglandin, and vasopressin.
  • PACAP pituitary adenylate cyclase- activating peptide
  • exemplary G-protein activators for use in the methods and compositions of the invention include, but are not limited to, cholera toxin or a subunit thereof and pertussis toxin or a subunit thereof.
  • adenylate cyclase activators for use in the methods and compositions of the invention include the pyrazole derivative A02011-1 [55] and benzyloxybenzaldehyde and analogs thereof [56].
  • PDE Inhibitors include the pyrazole derivative A02011-1 [55] and benzyloxybenzaldehyde and analogs thereof [56].
  • a cAMP elevator for use in the methods and compositions of the present invention can be a PDE inhibitor.
  • Cyclic nucleotide phosphodiesterases are enzymes that regulate the cellular levels of the second messengers, cAMP and cGMP, by controlling their rates of degradation.
  • PDE1 cAMP/cGMP
  • PDE2 cAMP/cGMP
  • PDE3 cAMP>>cGMP
  • PDE4 cAMP
  • PDE5 cGMP
  • PDE6 cGMP
  • PDE7 cAMP
  • PDE8 cAMP
  • PDE9 cGMP
  • PDE10 cAMP/cGMP
  • PDE11 cAMP/cGMP
  • nonspecific and selective or partially selective PDE inhibitors are known and can be used within the methods and compositions of the present invention.
  • the non-specific PDE inhibitor 3-Isobutyl-1- methylxanthine (IBMX)
  • IBMX 3-Isobutyl-1- methylxanthine
  • Other non-specific PDE inhibitors include, but are not limited to, theophylline, theobromine, aminophylline, pentoxifylline, and caffeine and other methyl xanthine and non-xanthine derivatives.
  • Selective or partially selective PDE inhibitors for use in the methods and compositions of the invention include, but are not limited to, Vinpocetine (e.g., INTELECTOL®) (available from, e.g., Covex Pharma Inc., Miami, Florida); Nicardipine HCl (available from, e.g., Par Pharmaceutical Companies, Inc., Spring Valley, New York); 8-MeOM-IBMX (8- methoxymethyl-3-isobutyl-1-methylxanthine) (available from Biomol International LP, Plymouth Meeting, Pennsylvania); EHNA (erythro-9-(2- hydroxy-3-nonyl)adenine) (available from, e.g., A.G.
  • Vinpocetine e.g., INTELECTOL®
  • Nicardipine HCl available from, e.g., Par Pharmaceutical Companies, Inc., Spring Valley, New York
  • 8-MeOM-IBMX 8- methoxymethyl-3-isobutyl-1-methylxant
  • Sildenafil citrate e.g., VIAGRA®, available from Pfizer, Inc., New York, New York
  • Zaprinast available from, e.g., A.G.
  • a PDE inhibitor for is a cAMP-specific PDE inhibitor.
  • the cAMP-specific inhibitor is a PDE3 inhibitor, a PDE4 inhibitor, a PDE7 inhibitor, or a PDE8 inhibitor, including, but not limited to, compounds described above and summarized in Table 1.
  • PDE3 is significantly expressed in cardiac and vascular myocytes, brain, and liver but not in skeletal muscle [30,31].
  • PDE4 is expressed widely in most tissues but is the predominant PDE isoenzymes in skeletal muscle and most immune cells. PDE4 is also present at relatively high levels in liver, heart, brain, smooth muscle, and vascular endothelium cells [30,31,32,33,34,35].
  • PDE7 is highly expressed in immune system, skeletal muscle, heart, liver, and also found in brain, kidney, and lung [30,31].
  • PDE8 is expressed in testes, cardiomyocytes, thyroid gland , brain, and adrenal gland [36].
  • the cAMP-specific PDE inhibitor can be a PDE4 inhibitor.
  • the PDE4 family encompasses four subtypes, which are designated PDE4 A, PDE4 B, PDE4 C, and PDE4 D and differ in their regulatory behavior and tissue expression patterns.
  • PDE4 inhibitors exhibit structural diversity and include compounds as described above in Table 1, as well as xanthine derivatives, such as arofylline (available from Almirall Prodesfarma, S.A.) and cipamfylline (GlaxoSmithKline, Research Triangle Park, North Carolina); catechol derivatives, such as rolipram (EMD Biosciences, San Diego, California), Ro 20-1724 (A.G.
  • PDE4 inhibitors are described, for example, in [60], which is incorporated herein by reference in its entirety.
  • PDE4 inhibitors that can be used within the methods and compositions of the invention include CC-10015 (available from Celgene Corporation), 4AZA-PDE4i (available from Elbion NV), ELB353 (available from Elbion NV), ELB326 (available from Elbion NV), GRC 4039 (available from Glenmark Pharmaceuticals Limited), GRC 4039 (available from Glenmark Pharmaceuticals Limited), IPL4088 (available from Inflazyme Pharmaceuticals Ltd.), MEM 1917 (available from Memory Pharmaceuticals Corp), PLX369 / PDE 4 Inhibitor (available from Plexxikon Inc.), AVE8112 (available from Sanofi-Aventis), Theophylline (available from SCOLR Pharma Inc), Oglemilast (available from Teijin Pharma Limited), Oglemilast / GRC 3886 (available from Teijin Pharma Limited), Z15370A (available from Zambon Group), LAS 37779 (available from Almirall Prodesfarma, S.A.), Atopik (
  • Drotaverine hydrochloride available from ELSaad Pharmaceutical Industries
  • Theophylline available from Eurand
  • Puroxan available from Eurodrug Laboratories
  • Choledyl Choline theophyllinate, theophylline) (available from Galenica s.a.)
  • Drotaverine- Grindeks available from Grindeks
  • Tromphylin theophylline
  • Hesotanol etophylline nicotinate
  • Neophin diethylaminoethyl theophylline
  • Arutopa acepifylline
  • Theophylline available from Indchemie Health Specialities Pvt.
  • Theophylline and Etophylline available from Indchemie Health Specialities Pvt. Ltd), Doverin (drotaverine) (available from Intas Pharmaceuticals Ltd.), Euphyllinum-N (theophylline) (available from JSC Farmak International), Theophar (theophylline) (available from Julphar), Draw (drotaverine) (available from Kamron Laboratories Ltd.), Quibron-T (theophylline) (available from King Pharmaceuticals Inc), Quibron-T/SR (theophylline anhydrous) (available from King Pharmaceuticals Inc), Theodur (theophylline) / Theodrip (available from Kowa Co., Ltd.), Teotard (theophylline) (available from Krka, d.
  • Theolan-B SR (theophylline) (available from KunWha Pharmaceutical Co., Ltd.), Hespil (acepifylline) (available from Kyung Dong Pharma. Co., Ltd.), Theophylline monohydrate (available from Laboratoires SMB SA), Sedacris (theophylline, guaifenesin) (available from Laboratorio Elea SACIFYA), Aminofilin (theophyllin) (available from Laboratorios Phoenix), Dexa aminofilin (dexamethasone and theophylline) (available from Laboratorios Phoenix), Dexa teosona (dexamethasone and theophylline) (available from Laboratorios Phoenix), Inastmol (ketotifen and theophylline) (available from Laboratorios Phoenix), Teosona (theophylline) (available from Laboratorios Phoenix), Theodur (theophylline) (available from Lavipharm Group),
  • Drotikind (drotaverine hydrochloride) (available from Mankind Pharma Ltd.), Ranispas-DV (drotaverine, omeprazole hydrocholoride) (available from Mankind Pharma Ltd.), Drot (drotaverine hydrochloride) (available from Mapra Laboratories Pvt.
  • a combination of PDE4 inhibitors and PDE7 inhibitors can be used for correcting glycogen storage in skeletal muscle and the brain.
  • a combination of PDE4 and PDE3 inhibitors can be combined to enhance the therapeutic efficacy in the liver and heart.
  • the cAMP elevator for use in the methods and compositions of the present invention is a Toll-like receptor ligand.
  • Toll-like receptors are a class of single membrane-spanning non-catalytic receptors that recognize structurally conserved molecules derived from microbes once they have breached physical barriers such as the skin or urinary tract mucosa and activate immune cell responses.
  • the Toll-like receptor family has been described as type I transmembrane pattern recognition receptors that possess varying numbers of extracellular N-terminal leucine-rich repeat motifs, followed by a cysteine-rich region, a TM domain, and an intracellular Toll/IL-1 R (TIR) motif. [61-67].
  • the leucine-rich repeat domain is important for ligand binding and associated signaling and the TIR domain is important in protein- protein interactions and is typically associated with innate immunity. [68-71].
  • the human TLR family is composed of at least 10 members, each of which is specific in its expression patterns and pathogen-associated molecular pattern sensitivities. [72-73].
  • Toll-like receptor ligands that activate the TLR pathway thus represent other cAMP elevators useful in the present invention.
  • Exemplary Toll-like receptor ligands for use within the methods and compositions of the invention include, but are not limited to, lipopolysaccharide (LPS), 1-palmitoyl-2- linoleoyl-sn-glycero-3-phosphocholine (pLPC), lipoteichoic acid (LTA), and flagellin.
  • LPS lipopolysaccharide
  • pLPC 1-palmitoyl-2- linoleoyl-sn-glycero-3-phosphocholine
  • LTA lipoteichoic acid
  • the cAMP elevator for use within the methods and compositions of the invention is a calcium ionophore.
  • Calcium ionophores act as calcium activators and include, but are not limited to, ionomycin calcium salts (Sigma) or A23187 (Sigma) (see also, [74-75]).
  • the cAMP elevator for use within the methods and compositions of the invention is an activator of protein kinase A.
  • Suitable protein kinase A (PKA) activators include, but are not limited to, 6-Bnz-cAMP, 8-CPT-2’-O-Me-cAMP, 8-CPT-cAMP, 8-Bromo-cAMP, Dibutyryl-cAMP, Dioctanoyl-cAMP, Sp-8-Br-cAMPS, Sp-cAMPS, cAMP, and a PKA subunit.
  • PKC protein kinase C
  • PKC protein kinase C
  • PKC diacylglycerol
  • PLC phospholipase C
  • PLA 2 phospholipase A 2
  • IP 3 inositol-1,4,5-trisphosphate
  • PKC activators potentiate forskolin-induced cAMP formation.
  • the PKC activator for use within the methods and compositions of the invention is phorbol myristate acetate (PMA) or a PKC purified enzyme.
  • Beta2-adrenergic agonists also known as beta2-adrenergic receptor agonists, act on beta2-adrenergic receptors.
  • ⁇ adrenergic receptors are coupled to a stimulatory G protein of adenylyl cyclase. This enzyme produces the second messenger cyclic adenosine monophosphate(cAMP). beta2- adrenergic agonists therefore can increase cAMP production
  • beta2-adrenergic agonists include, for example, bitolterol, fenoterol, isoprenaline, levosalbutamol, orciprenaline, pirbuterol, procaterol, ritodrine, salbutamol, terbutaline, arformoterol, bambuterol, clenbuterol, formoterol, salmeterol, indacaterol, olodaterol, vilanterol, vilanterol with umeclidinium bromide, vilanterol with fluticasone furoate, zilpaterol.
  • Adenylate cyclase toxin represents another type of cAMP elevator for use in the methods and compositions of the present invention.
  • Adenylate cyclase toxin is a single polypeptide A/B-type bacterial toxin that has the ability to interact with target cells, insert into the cytoplasmic membrane, and deliver its adenylate cyclase enzymatic domain to the cell interior [84-85]. Once entry has occurred, the enzymatic activity of the toxin produces cAMP from host cell ATP [86]. Accordingly, a further cAMP elevator that can be used in the methods and compositions of the present invention is adenylate cyclase toxin.
  • treat and treatment refer to the application or administration of one or more cAMP elevators or agents that mimic cAMP to an individual having one of the GSDs or other conditions where there is a build of glycogen.
  • treat and treatment also refer to amelioration of one or more symptoms associated with the diseases, prevention or delay of the onset of one or more symptoms of the diseases, and/or lessening of the severity or frequency of one or more symptoms of the diseases.
  • treatment can refer to improvement of liver (e.g., improvement of liver enzymes, prevention of the progressive fibrosis, reduction of liver size, and stabilization of the disease); improvement of muscle function (e.g., prevention of progressive myopathy, increase in muscle strength, increase in function and activities of daily living); improvement of cardiac status (e.g., prevention of ventricular hypertrophy, cardiomyopathy, and rhythm disturbances); improvement in neurodevelopment and/or motor skills (e.g., increase in AIMS score and functional measures such as 6MWT); reduction of glycogen levels in tissue of the individual affected by the diseases; or any combination of these effects.
  • liver e.g., improvement of liver enzymes, prevention of the progressive fibrosis, reduction of liver size, and stabilization of the disease
  • improvement of muscle function e.g., prevention of progressive myopathy, increase in muscle strength, increase in function and activities of daily living
  • improvement of cardiac status e.g., prevention of ventricular hypertrophy, cardiomyopathy, and rhythm disturbances
  • improvement in neurodevelopment and/or motor skills
  • improve indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control individual (or multiple control individuals) in the absence of the treatment described herein.
  • a control individual is an individual afflicted with the same type/form and stage of GSD as the individual being treated, who is about the same age as the individual being treated (to ensure that the stages of the disease in the treated individual and the control individual(s) are comparable).
  • a value relative to baseline can improve or increase by about 3, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100% or more.
  • a value relative to baseline can improve or decrease by about 3, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100% or more.
  • the individual being treated can be an individual (fetus, child, adolescent, or adult human) having a certain type of GSDs (i.e., type I, II, III, IV, V, VI, VII, IX, XI, XII, XIII, XIV, Lafora disease, Fanconi-Bickel disease, Danon disease, PRKAG2 cardiac syndrome, etc.).
  • GSDs i.e., type I, II, III, IV, V, VI, VII, IX, XI, XII, XIII, XIV, Lafora disease, Fanconi-Bickel disease, Danon disease, PRKAG2 cardiac syndrome, etc.
  • An individual can also have a condition where there is a secondary build-up of glycogen.
  • Such conditions include, for example, Niemann-Pick Disease [87].
  • Niemann-Pick Disease encompasses a group of lysosomal storage diseases that affect metabolism and are caused by genetic mutations.
  • GSD X phosphoglycerate mutase deficiency; increased glycogen in muscle
  • phosphoglycerate kinase deficiency increased glycogen in muscle
  • RBCK1 deficiency polyglucosan body myopathy caused by deficiency of ubiquitin ligase RBCK1; polyglucosan body
  • GSD XV Glycogenin-1 deficiency; polyglucosan body
  • the individual can have residual (partial loss) enzyme activity, or no measurable (complete loss) enzyme activity.
  • subject refers to any organism to which the presently disclosed treatment methods and pharmaceutical compositions can be administered.
  • a subject is a mammal.
  • a subject is a primate, a human, a domestic animal, or an agricultural animal.
  • a subject can include a human subject for medical purposes, such as treatment of a condition or disease, or an animal subject for medical, veterinary purposes, or developmental purposes. Suitable animal subjects include mammals and avians.
  • the term“avian” as used herein includes, but is not limited to, chickens, ducks, geese, quail, turkeys, and pheasants.
  • mammal includes, but is not limited to, primates, e.g., humans, monkeys, apes, and the like; bovines, e.g., cattle, oxen, and the like; ovines, e.g., sheep and the like; caprines, e.g., goats and the like; porcines, e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys, zebras, and the like; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, and the like; and rodents, including mice, rats, and the like.
  • the subject is a mammal such as a domestic cat or dog.
  • the subject is a human.
  • the term subject and patient are used interchangeably herein.
  • the therapeutically effective amount of a composition or medicament can be administered at regular intervals, depending on the nature and extent of the disease's effects, and on an ongoing basis.
  • Administration at a "regular interval,” as used herein, indicates that the therapeutically effective amount is administered periodically (as distinguished from a one-time dose).
  • the interval can be determined by standard clinical techniques.
  • a cyclic AMP elevator can be administered monthly, every two weeks, weekly, twice weekly, daily, twice daily, three times daily or more.
  • the administration interval for a single individual need not be a fixed interval, but can be varied over time, depending on the needs of the individual.
  • compositions and Kits The cyclic AMP elevators can be used to make pharmaceutical compositions.
  • Pharmaceutical compositions comprising the one or more (e.g., 1, 2, 3, 4, 5, 6, or more) of the cAMP elevators described above are provided and can include a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier is any carrier suitable for in vivo administration. Examples of pharmaceutically acceptable carriers suitable for use in the composition include, but are not limited to, water, buffered solutions, glucose solutions, oil-based or bacterial culture fluids. Additional components of the compositions can suitably include, for example, excipients such as stabilizers, preservatives, diluents, emulsifiers and lubricants.
  • Examples of pharmaceutically acceptable carriers or diluents include stabilizers such as carbohydrates (e.g., sorbitol, mannitol, starch, sucrose, glucose, dextran), proteins such as albumin or casein, protein-containing agents such as bovine serum or skimmed milk and buffers (e.g., phosphate buffer). Especially when such stabilizers are added to the compositions, the composition is suitable for freeze-drying or spray-drying. The composition can also be emulsified.
  • carbohydrates e.g., sorbitol, mannitol, starch, sucrose, glucose, dextran
  • proteins such as albumin or casein
  • protein-containing agents such as bovine serum or skimmed milk
  • buffers e.g., phosphate buffer
  • compositions including two or more cAMP elevators are provided herein.
  • the two or more cAMP elevators within the pharmaceutical composition can be from the same class (or type) of cAMP elevators; in other embodiments, the two or more cAMP elevators can be from two or more classes of cAMP elevators.
  • the two or more cAMP elevators within the pharmaceutical composition are thus selected from one or more of the following non-limiting examples of classes of cAMP elevators: adenylate cyclase activators, PDE inhibitors, Toll-like receptor ligands, calcium ionophores, beta2-adrenergic receptor agonists, protein kinase A activators, protein kinase C activators, and adenylate cyclase toxin.
  • composition comprising two or more cAMP elevators comprises a combination of two or more adenylate cyclase activators
  • the adenylate cyclase activators can be selected from the group consisting of the labdane diterpenes in one embodiment.
  • at least one of the adenylate cyclase activators is a labdane diterpene
  • the remaining adenylate cyclase activator(s) is (are) selected from the group consisting of a G-protein coupled receptor agonist, a G-protein activator, the pyrazole derivative A02011-1 [48], and benzyloxybenzaldehyde and analogs thereof such as those disclosed in [49].
  • the composition comprises two or more cAMP elevators in therapeutically effective amounts for treating a GSD, and at least one of the cAMP elevators is an adenylate cyclase activator, and at least one of the remaining cAMP elevator(s) is a PDE inhibitor.
  • composition comprises two or more cAMP elevators
  • at least one of the cAMP elevators is an adenylate cyclase activator
  • at least one of the remaining cAMP elevator(s) is a Toll-like receptor ligand, a calcium ionophore, a protein kinase A activator, a protein kinase C activator, a beta2-adrenergic receptor agonists, or adenylate cyclase toxin.
  • One or more cAMP elevators can also be administered with one or more enzyme replacement therapies, gene therapies, chaperone therapies, or substrate reduction therapies using siRNA/shRNA, anti-sense oligonucleotides, or small molecule or peptide drugs.
  • An enzyme replacement therapy is any therapy with the purpose of replacing or overcoming an enzyme deficiency in a subject.
  • Protein based therapies in which one or more enzymes are provided to the subject or to cells in the subject directly is an enzyme replacement therapy.
  • GSD II can be treated by administration of acid alpha-glucosidase.
  • the enzyme replacement therapy can be or can include, but is not limited to, administration of acid alpha-glucosidase, glucose-6- phosphatase, glycogen debranching enzyme, glycogen branching enzyme, muscle glycogen phosphorylase, liver glycogen phosphorylase, muscle phosphofructokinase, phosphorylase kinase, glucose transporter, Aldolase A, phosphoglucomutase deficiency, LAMP-2, ⁇ -enolase, ⁇ -glucuronidase, imiglucerase, agalsidase alpha, agalsidase beta, aglucosidase alpha, laronidase, idursulphase, galsulphase, or combinations thereof.
  • the enzyme replacement therapy and the cAMP elevator can be delivered in a single pharmaceutical composition or can be administered in separate compositions.
  • a chaperone therapy can be added into the single pharmaceutical composition or can be administered as a separate composition.
  • Gene therapies include, for example, administration of a therapeutic nucleic acid encoding a functional GAA (for GSD II), G6Pase (for GSD I), GBE (for GSD IV), GDE (for GSD III), or a deficient enzyme or protein in other types of GSDs.
  • the therapeutic nucleic acid can be delivered, for example, by a viral vector, such as adenoviruses, adeno-associated viruses (AAVs), lentiviruses, herpes virus, pox virus, human foamy virus (HFV), or retroviruses, or by a nonviral delivery platform, such as naked DNA vector, lipid/polymer- or nanoparticle-based nucleic acid delivery systems, etc.
  • a viral vector such as adenoviruses, adeno-associated viruses (AAVs), lentiviruses, herpes virus, pox virus, human foamy virus (HFV), or retroviruses
  • AAVs adeno-associated viruses
  • lentiviruses lentiviruses
  • herpes virus herpes virus
  • pox virus pox virus
  • human foamy virus HBV
  • retroviruses retroviruses
  • a nonviral delivery platform such as naked DNA vector, lipid/polymer- or nanoparticle
  • Methods of inhibiting glycogen synthase include, but are not limited to an RNAi-based therapy using small interference RNA (siRNA) or short hairpin RNA (shRNA), an antisense therapy using an anti-sense oligonucleotide (ASO), or therapies using small molecules or peptides.
  • siRNA small interference RNA
  • shRNA short hairpin RNA
  • ASO anti-sense oligonucleotide
  • therapies using small molecules or peptides include, but are not limited to an RNAi-based therapy using small interference RNA (siRNA) or short hairpin RNA (shRNA), an antisense therapy using an anti-sense oligonucleotide (ASO), or therapies using small molecules or peptides.
  • shRNA mediated gene silencing of glycogen synthase (GYS) and glycogenin (GYG) the two major enzymes involved in glycogen synthesis, led to a decrease in cytoplasmic and lysosomal glyco
  • Small molecule inhibitors such as small molecule inhibitors of glycosyltransferases can be used in treatment of type 1 Gaucher disease.
  • Inhibitors of glucosylceramide biosynthesis can be used to treat Gaucher disease.
  • siRNAs, shRNAs, anti-sense oligonucleotides, small molecules and peptides that target glycogen synthase are known in the art.
  • a chaperone therapy provides molecules that can assist in the folding of enzymes or proteins (e.g., enzymes and proteins provided as part of an enzyme replacement therapy or gene therapy).
  • the chaperone therapy molecules can also help the enzymes or proteins retain their catalytic activity, prevent their recognition by quality control systems in cells that can destroy the enzymes or proteins, and provide improved trafficking of the enzymes or proteins to their final destination.
  • Examples of chaperone therapy include, for example, the use of 1-deoxy-galactonojirimycin (DGJ), to enhance, for example, alpha-galactosidase activity in Fabry disease.
  • DGJ 1-deoxy-galactonojirimycin
  • chaperone therapies include, for example, dimethyl sulfoxide and trimethylamine N-oxide, galactose, N-(n-nonyl)deoxynorjirimycin, N-(n-butyl)deoxynojirimycin, deoxynojirimycin, N370S and G202R GC pharmacologic chaperones, N-octyl- isofagomine, N-octyl-2,5-dideoxy-2,5-imino-D-glucitol, ⁇ -1-C-nonyl-1,5- dideoxy-1,5-imino-D-xylitol, isofagomine, adamantyl terminated N-alkyl isofagomines, 2,5-anhydro-2,5-imino-D-glucitol derativatives, N-adamantyl-4- ((3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)
  • a cAMP elevator (or more than one cAMP elevator) and an enzyme replacement therapy or a gene therapy, or chaperone therapy, a substrate reduction therapy can take many forms.
  • the compositions can be administered in any order, at the same time or as part of a unitary composition.
  • the two or more compositions can be administered such that one composition is administered before the other with a difference in administration time of 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, 1 day, 2 days, 4 days, 7 days, 2 weeks, 4 weeks or more.
  • compositions for treating GSD or other conditions where there is a build of glycogen in a subject in need thereof that include two or more cAMP elevators in therapeutically effective amounts for treating a GSD or other condition where there is a build of glycogen, and can include a pharmaceutically acceptable carrier.
  • this pharmaceutical composition further comprises one or more additional enzyme replacement therapies or chaperone therapies.
  • the present invention relates to a pharmaceutical composition for treating a disease or condition in a subject in need thereof that includes at least one cAMP elevator and one or more enzyme replacement therapies or chaperone therapies each of which is present in a therapeutically effective amount for treating a disease or condition in a subject in need thereof, and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.
  • an effective amount or a therapeutically effective amount as used herein means the amount of a compound that, when administered to a subject for treating GSD or other conditions where there is a build of glycogen is sufficient to effect a treatment (as defined above).
  • the therapeutically effective amount will vary depending on the compounds, formulation or composition, the disease and its severity and the age, weight, physical condition and responsiveness of the subject to be treated. One of skill in the art can determine a therapeutically effective amount.
  • compositions described herein can be administered by any means known to those skilled in the art, including, but not limited to, oral, topical, intranasal, intradermal, intraperitoneal, parenteral, intravenous, intramuscular, subcutaneous, intrathecal, transcutaneous, nasopharyngeal, or transmucosal absorption. Therefore, the compounds can be formulated as an ingestible, injectable, topical or suppository formulation. The compounds can also be delivered with in a liposomal or time-release vehicle. Administration of the compounds to a subject in accordance with the invention can exhibit beneficial effects in a dose-dependent manner. Thus, within broad limits, administration of larger quantities of the compounds is expected to achieve increased beneficial biological effects than administration of a smaller amount. Moreover, efficacy is also contemplated at dosages below the level at which toxicity is seen.
  • the specific dosage administered in any given case will be adjusted in accordance with the compound or compounds being administered, the disease to be treated or inhibited, the condition of the subject, and other relevant medical factors that can modify the activity of the compound or the response of the subject, as is well known by those skilled in the art.
  • the specific dose for a particular subject depends on age, body weight, general state of health, diet, the timing and mode of administration, the rate of excretion, medicaments used in combination and the severity of the particular disorder to which the therapy is applied. Dosages for a given patient can be determined using conventional considerations, e.g., by customary comparison of the differential activities of the compound of the invention and of a known agent, such as by means of an appropriate conventional pharmacological or prophylactic protocol.
  • the maximal dosage for a subject is the highest dosage that does not cause undesirable or intolerable side effects.
  • the number of variables in regard to an individual prophylactic or treatment regimen is large, and a considerable range of doses is expected.
  • the route of administration will also impact the dosage requirements. It is anticipated that dosages of the compound will reduce symptoms of the condition at least 5%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to pre-treatment symptoms or symptoms is left untreated. It is specifically contemplated that pharmaceutical preparations and compositions can palliate or alleviate symptoms of the disease without providing a cure, or, in some embodiments, can be used to cure the disease or disorder.
  • a therapeutically effective amount of a cAMP elevator or additional active compound within the methods and compositions of the present invention typically ranges from about 1 ⁇ g/kg to about 500 mg/kg, about 10 ⁇ g/kg to about 500 mg/kg, about 100 ⁇ g/kg to about 500 mg/kg, about 1 mg/kg to about 500 mg /kg, about 1 mg/kg to about 400 mg/kg, about 1 mg/kg to about 300 mg/kg, about 1 mg/kg to about 200 mg/kg, about 1 mg/kg to about 100 mg/kg, about 1 mg/kg to about 75 mg/kg, about 1 mg/kg to about 50 mg/kg, or about 1 mg/kg to about 25 mg/kg.
  • the therapeutically effective dose of a cAMP elevator or additional active compound is an amount of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg, about 6 mg/kg, about 7 mg/kg, about 8 mg/kg, about 9 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg, about 45 mg/kg, about 50 mg/kg, about 55 mg/kg, about 60 mg/kg, about 65 mg/kg, about 70 mg/kg, about 75 mg/kg, about 80 mg/kg, about 85 mg/kg, about 90 mg/kg, about 95 mg/kg, about 100 mg/kg, about 110 mg/kg, about 120 mg/kg, about 125 mg/kg, about 130 mg/kg, about 140 mg/kg, about 150 mg/kg, about 160 mg/kg, about 170 mg/kg, about 175 mg/kg, about 180 mg/kg,
  • the therapeutically effective amount can be even lower, for example from about 1 ng/kg to about 1 mg/kg, about 50 ng/kg to about 1 mg/kg, about 100 ng/kg to about 1 mg/kg, about 500 ng/kg to about 1 mg /kg, about 1 ⁇ g/kg to about 1 mg/kg, about 50 ⁇ g/kg to about 1 mg/kg, about 100 ⁇ g/kg to about 1 mg/kg, or about 500 ⁇ g/kg to about 1 mg/kg.
  • the therapeutically effective dose of a cAMP elevator or additional active compound is an amount of about 1 ng/kg, about 5 ng/kg, about 10 ng/kg, about 20 ng/kg, about 30 ng/kg, about 40 ng/kg, about 50 ng/kg, about 100 ng/kg, about 200 ng/kg, about 300 ng/kg, about 400 ng/kg, about 500 ng/kg, about 600 ng/kg, about 700 ng/kg, about 800 ng/kg, about 900 ng/kg, about 1 ⁇ g/kg, about 5 ⁇ g/kg, about 10 ⁇ g/kg, about 20 ⁇ g/kg, about 30 ⁇ g/kg, about 40 ⁇ g/kg, about 50 ⁇ g/kg, about 100 ⁇ g/kg, about 200 ⁇ g/kg, about 300 ⁇ g/kg, about 400 ⁇ g/kg, about 500 ⁇ g/kg, about 600 ⁇ g/kg, about 700 ⁇ g/kg, about 800 ⁇ g
  • Kits comprising one or more cyclic AMP elevators (e.g., about 1, 2, 3, 4, 5, 6, or more), one or more enzyme replacement therapies or gene therapies (e.g., 1, 2, 3, 4, 5, 6, or more) and instructions for administering the cyclic AMP elevator and the enzyme replacement therapy to a subject with a glycogen storage disease or condition with a buildup of glycogen are also provided.
  • the kits can additionally include one or more chaperone therapies (e.g., 1, 2, 3, 4, 5, 6, or more).
  • the enzyme replacement therapy included in these kits replaces an enzyme deficient in the glycogen storage disease.
  • Kits comprising at least one cyclic AMP elevator (e.g., about 1, 2, 3, 4, 5, 6, or more), one or more substrate reduction therapies (e.g., about 1, 2, 3, 4, 5, 6, or more), and instructions for administering the cyclic AMP elevator and the substrate reduction therapy to a subject with a glycogen storage disease or condition with a buildup of glycogen are also provided.
  • cyclic AMP elevator e.g., about 1, 2, 3, 4, 5, 6, or more
  • substrate reduction therapies e.g., about 1, 2, 3, 4, 5, 6, or more
  • instructions for administering the cyclic AMP elevator and the substrate reduction therapy to a subject with a glycogen storage disease or condition with a buildup of glycogen are also provided.
  • Example 2 Forskolin treatment for GSD IV.
  • Fibroblasts from a patient with GSD IV were used to evaluate the in vitro efficacy of Forskolin treatment.
  • Fibroblast cells were grown to confluency in DMEM containing 10% FBS [41].
  • Forskolin was added to the culture medium at a final concentration of 1 ⁇ M (low-dose) or 10 ⁇ M (high-dose). After 18 h, the cells were washed three times with cold phosphate buffered saline and then collected with a scraper. Glycogen content was determined in the cell lysates.
  • Example 3 In vitro screening of cAMP elevator drugs
  • a cAMP activator drug will induce glycogen degradation and inhibit glycogen synthesis via PKA activation, and hence reduce cytoplasmic glycogen accumulation in the affected tissues of, for example, GSD III and IV patients.
  • PDE inhibitor drugs can be evaluated for treatment of GSD IV and GSD III using cellular and animal disease models.
  • Mouse C2C12 or rat L6 muscle cells and human HepG2 or mouse AML12 liver cells can be used as in vitro platforms for screening cAMP elevator by assessing cAMP changes in muscle and liver, respectively.
  • a broad concentration curve for each compound can be determined and cAMP levels can rapidly be quantified in the cells or media (or both) at different time points.
  • candidate PDE inhibitor drugs i.e., PDE4 inhibitors Crisaborole, E6005 (RVT 501), roflumilast (Daliresp), apremilast (Otezla), etc; PDE3 inhibitors Cilostazol, Pletal (cilostazol), Perfan I.V. (enoximone), Primacor (minnone lactate) etc.; PDE7 inhibitors BRL50481, IC242, ABS16165, etc.
  • PDE7 inhibitors BRL50481, IC242, ABS16165, etc. can be screened in vitro using C2C12 or HepG2 cells seed
  • each compound can be tested in C2C12 myoblast cells (for screening of PDE4 and PDE7 inhibitors) or HepG2 liver cells (for screening of PDE3 and PDE4 inhibitors).
  • C2C12 myoblast cells for screening of PDE4 and PDE7 inhibitors
  • HepG2 liver cells for screening of PDE3 and PDE4 inhibitors.
  • both media and cells can be collected at different time points (i.e. 30 min, 2 h, 6 hr, 12 hr, and 24 hr) to quantify cAMP levels using standard colorimetric cAMP kits.
  • Forskolin can be used as positive control.
  • Data from the experiment can be used to determine the most effective dose and responsive time of each compound. Based upon the ability of each candidate drug to induce cAMP elevation, several compounds from each class of PDE4, PDE3, and PDE7 inhibitors can be selected for further evaluation.
  • Example 4 Examination of the ability of PDE4 inhibitors to elevate cAMP and reduce glycogen storage in primary GSD patient fibroblast cells It has been reported that PDE4s, but not PDE3 and PDE7, are the dominant isoforms in human fibroblasts [42]. Primary fibroblast cells derived from patients with GSD IV, GSD III, and other GSDs can be tested.
  • the most effective dose to elevate cellular cAMP can be used for treatment of GSD IV ,GSD III, or other types of GSD patient fibroblasts.
  • Group 2. Forskolin treatment group add 1 ⁇ M Forskolin (positive control); Group 3. PDE4 inhibitor at the most effective dose.
  • GSD patient fibroblast cells can be grown to confluency in DMEM containing 10% FBS.
  • the tested drug can be added to culture medium at the indicated concentration for each group. After 12, 24, or 48 hr, cells can be washed three times with cold PBS buffer and collected with a scraper. Glycogen content and cAMP level can be assayed in cell lysate.
  • primary GSD IIIa patient myoblast cells can be used to test the ability of PDE4 inhibitor or PDE7 inhibitor (or both) to reduce glycogen content.
  • PDE4 inhibitor treatment group human equivalent dose. Treatment can start at the age of about 2 months. All mice can be euthanized at age 5 months following overnight fasting, to collect urine, blood, and tissues. Dosage regimen and administration route will be determined based on human use of each drug. Glycogen content can be analyzed in different tissues including liver, heart, skeletal muscles, brain, and diaphragm, as described in, for example, [40,43]. Tissue histology can be analyzed in a pathology laboratory. Urine can be used for testing urinary Hex4, a biomarker for several GSDs, by stable isotope-dilution electrospray tandem mass spectrometry as previously described in [44].
  • Blood chemistry including AST, ALT, ALP, CPK, GLU, etc. can be analyzed [45]. Behavioral and muscle function can be tested at about ages 2, 3.5, and 5 months, to assess reversal of neuromuscular involvement by treadmill, Rota-rod, wire-hang tests as described in, for example, [43,46] and.
  • Combined therapy with inhibitors of PDE4 + PDE3 (for liver) or PDE4 + PDE7 (for muscle) can also be tested in GSD IV or GSD III mice.

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Abstract

L'invention concerne des compositions et des méthodes pour le traitement de glycogénoses ou d'états pathologiques apparentés révélant une accumulation de glycogène. Les compositions élevant la teneur en AMP cyclique se sont avéré réduire le stockage du glycogène dans les cellules atteintes et peuvent par conséquent être utilisées pour traiter ou réduire les symptômes chez des sujets présentant des glycogénoses ou des états pathologiques apparentés révélant une accumulation de glycogène.
PCT/US2016/025215 2015-03-31 2016-03-31 Méthodes et compositions pour le traitement de glycogénoses à l'aide d'agents imitant l'amp cyclique ou élevant celui-ci WO2016161086A1 (fr)

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017120420A1 (fr) * 2016-01-06 2017-07-13 The Trustees Of Columbia University In The City Of New York Utilisation de gaïacol pour la prévention et le traitement de glycogénoses
WO2018154578A1 (fr) * 2017-02-22 2018-08-30 Hadasit Medical Research Services And Development Ltd. Composés pour le traitement de troubles de stockage du glycogène
CN110914419A (zh) * 2017-03-10 2020-03-24 吉尼松公司 糖原贮积病iii的治疗
EP3471781A4 (fr) * 2016-06-17 2020-05-06 Ionis Pharmaceuticals, Inc. Modulation de l'expression de gys1
WO2020210798A1 (fr) * 2019-04-12 2020-10-15 Loma Linda University Méthodes de traitement de la maladie de niemann-pick de type c
WO2020225535A1 (fr) * 2019-05-08 2020-11-12 University Of Newcastle Upon Tyne Dosages de criblage de kinase
US20200405722A1 (en) * 2017-05-12 2020-12-31 Duke University Methods for the Use of Low-Dose Immune Modulators Transiently for Treating Patients Undergoing Protein Replacement Therapy

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008157205A2 (fr) * 2007-06-15 2008-12-24 Duke University Procédés et compositions pour le traitement des infections de l'appareil urinaire en utilisant des agents qui imitent ou augmentent l'amp cyclique
US8679478B2 (en) * 2010-10-04 2014-03-25 Duke University Methods of lysosomal storage disease therapy
WO2014130723A1 (fr) * 2013-02-20 2014-08-28 Valerion Therapeutics, Llc Méthodes et compositions pour le traitement de la maladie de pompe

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008157205A2 (fr) * 2007-06-15 2008-12-24 Duke University Procédés et compositions pour le traitement des infections de l'appareil urinaire en utilisant des agents qui imitent ou augmentent l'amp cyclique
US8679478B2 (en) * 2010-10-04 2014-03-25 Duke University Methods of lysosomal storage disease therapy
WO2014130723A1 (fr) * 2013-02-20 2014-08-28 Valerion Therapeutics, Llc Méthodes et compositions pour le traitement de la maladie de pompe

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KOEBERL, DD ET AL.: "Enhanced Efficacy of Enzyme Replacement Therapy in Pompe Disease Through Mannose-6-Phosphate Receptor Expression in Skeletal Muscle.", MOLECULAR GENETICS AND METABOLISM., vol. 103, no. 2, 2011, pages 107 - 112, 1-15, XP028216120 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10933031B2 (en) 2016-01-06 2021-03-02 The Trustees Of Columbia University In The City Of New York Use of guaiacol for the prevention and treatment of glycogen storage disease
WO2017120420A1 (fr) * 2016-01-06 2017-07-13 The Trustees Of Columbia University In The City Of New York Utilisation de gaïacol pour la prévention et le traitement de glycogénoses
US11236339B2 (en) 2016-06-17 2022-02-01 Ionis Pharmaceuticals, Inc. Modulation of GYS1 expression
EP3471781A4 (fr) * 2016-06-17 2020-05-06 Ionis Pharmaceuticals, Inc. Modulation de l'expression de gys1
US11713462B2 (en) 2016-06-17 2023-08-01 Ionis Pharmaceuticals, Inc. Modulation of GYS1 expression
US11053231B2 (en) 2017-02-22 2021-07-06 Hadasit Medical Research Services And Development Ltd. Compounds for the treatment of glycogen storage disorders
WO2018154578A1 (fr) * 2017-02-22 2018-08-30 Hadasit Medical Research Services And Development Ltd. Composés pour le traitement de troubles de stockage du glycogène
IL290955B (en) * 2017-02-22 2022-11-01 Univ Ramot Compounds for the treatment of glycogen storage diseases
IL290955B2 (en) * 2017-02-22 2023-03-01 Univ Ramot Compounds for the treatment of glycogen storage diseases
US11891381B2 (en) 2017-02-22 2024-02-06 Hadasit Medical Research Services And Development Ltd. Compounds for the treatment of glycogen storage disorders
CN110914419A (zh) * 2017-03-10 2020-03-24 吉尼松公司 糖原贮积病iii的治疗
US20200405722A1 (en) * 2017-05-12 2020-12-31 Duke University Methods for the Use of Low-Dose Immune Modulators Transiently for Treating Patients Undergoing Protein Replacement Therapy
US11944628B2 (en) 2017-05-12 2024-04-02 Duke University Methods for the use of low-dose immune modulators transiently for treating patients undergoing protein replacement therapy
WO2020210798A1 (fr) * 2019-04-12 2020-10-15 Loma Linda University Méthodes de traitement de la maladie de niemann-pick de type c
WO2020225535A1 (fr) * 2019-05-08 2020-11-12 University Of Newcastle Upon Tyne Dosages de criblage de kinase

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