EP1165063A1 - Utilisation d'acide succinique ou de ses sels dans le traitement du diabete et la cicatrisation de blessures - Google Patents

Utilisation d'acide succinique ou de ses sels dans le traitement du diabete et la cicatrisation de blessures

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Publication number
EP1165063A1
EP1165063A1 EP99933303A EP99933303A EP1165063A1 EP 1165063 A1 EP1165063 A1 EP 1165063A1 EP 99933303 A EP99933303 A EP 99933303A EP 99933303 A EP99933303 A EP 99933303A EP 1165063 A1 EP1165063 A1 EP 1165063A1
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EP
European Patent Office
Prior art keywords
succinic acid
pharmaceutically acceptable
acceptable salt
insulin
signal transduction
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.)
Withdrawn
Application number
EP99933303A
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German (de)
English (en)
Inventor
Igor Anatolievich Pomytkin
Olga Evgenievna Kolesova
Tatiyana Jurievna Ukhanova
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.)
Verteletsky Pavel Vasilievich
Original Assignee
Verteletsky Pavel Vasilievich
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Filing date
Publication date
Application filed by Verteletsky Pavel Vasilievich filed Critical Verteletsky Pavel Vasilievich
Publication of EP1165063A1 publication Critical patent/EP1165063A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/194—Carboxylic acids, e.g. valproic acid having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid
    • 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
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom

Definitions

  • the present invention relates to the use of succinic acid or a pharmaceutically acceptable salts thereof, and methods of inhibiting protein tyrosine phosphatases and treating disease states caused by dysfunctional signal transduction.
  • Protein tyrosine kinases are a family of transmembrane and cytoplasmic enzymes that specifically phosphorylate tyrosine residues in proteins. More than 50 receptors involved in regulation of cell growth, differentiation, and metabolism are known to have intrinsic protein tyrosine kinase activity. For example, the receptors to insulin, insulinlike growth factors (IGF), epidermal growth factor (EGF), transforming growth factor
  • TGF alpha 25 alpha
  • PDGF platelet-derived growth factor
  • FGF fibroblast growth factors
  • VEGF vascular endothelial growth factor
  • cytoplasmic tyrosine kinases like Src kinases (e.g., Src, Fyn), Janus kinases (e.g., JAK1, JAK2, TYK2).
  • External stimuli e.g. insulin, EGF, TGF alpha, and PDGF
  • tyrosine phosphorylation of the receptor itself and substrates e.g. insulin receptor substrate- 1 (LRS-1).
  • binding of external stimuli to receptors activates the associated tyrosine kinases to transduce a signal throughout the cell and into the nucleus.
  • the tyrosine phosphorylation was shown to be involved in gamma-interferon signaling mechanism that controls a diverse set of biological responses including antiviral protection and antiproliferation action, as well as plays a role in inflammation, tissue repair and host defense. Igarashi, K. et al., Mol. Cell. Biol. 13:1634-40 (1993); Igarashi, K. et al., Mol. Cell. Biol. 13:3984-9 (1993). The tyrosine phosphorylation was shown to be involved in propagation of alpha-interferon mediated signal response. David, M.
  • EGF receptor kinase activity was elevated more than 200% within 30 minutes, moreover, EGF receptor levels were increased by 36-fold within 24 hours after gastric mucosal injury and these increases were closely correlated with mucosal regeneration.
  • Relan N.K. et al., Lab. Invest. 73;717-26 (1996); Majumdar, NP. et al., J. Lab. Clin. Med. 128:173-80 (1996); Majumdar, A.P. et al., Am. J. Physiol. 274: G863-G870 (1998).
  • EGF receptor tyrosine kinase plays an essential role in regulating gastric mucosal cell proliferation in gastric ulcer healing. Tarnawski, A.S. et al, J. Clin. Gastroenterol. 27 Suppl. 1:S 12-20 (1998). It was reported that increased expression of EGF receptor in the epithelial cells of gastric ulcer margins and increased the receptor phosphorylation levels leading to a dramatic increase in MAP-ERK-1 and ERK-2 kinase phosphorylation levels more than 440% and 880% respectively during early stages of gastric ulcer healing. Moreover, tyrphostin A46, an inhibitor of EGF receptor kinase dependent cell proliferation, was reported to inhibit significantly both the signaling pathways and ulcer healing.
  • EGF and TGF alpha binding to a common EGF receptor tyrosine kinase initiates a series of events which ultimately regulate cell proliferation during repair of human burn wounds.
  • Insulin was reported to improve wound matrix formation in massively burned patients. Pierre, E.F. et al., J. Trauma 44:342-5 (1998).
  • Insulin and EGF was reported to accelerate gastric eithelial wound repair by stimulating both the migration and the proliferation of gastric epithelial cells.
  • PTPases Protein tyrosine phosphatases
  • the substrates of PTPases may be proteins which possess phosphotyrosine residues including PTKases (e.g. receptors to insulin, EGF, NEGF, TGF alpha, FGF, and PDGF) or substrates of PTKases (e.g. LRS-1).
  • PTKases e.g. receptors to insulin, EGF, NEGF, TGF alpha, FGF, and PDGF
  • substrates of PTKases e.g. LRS-1
  • PTPases are negative regulators of insulin and growth factors receptor signal transduction.
  • the PTP IB was reported to be a negative regulator of insulin and IGF-1 signal transduction.
  • the transmembrane PTPase LAR was shown to have a direct impact on insulin action. Goldstein, B.J. et al., Mol. Cell. Biochem. 182:91-9 (1998). Low molecular wight LMW PTPase was shown to be a negative regulator of insulin-mediated mitotic and metabolic signaling. Chiarugi, P. et al., Biochem. Biophys. Res. Commun. 238:676-82 (1997). As reported, PTPase LMW inhibits cell proliferation by dephosphorylation of the phosphorylated PDGF receptor. Chiarugi, P. et al., FEBS Lett. 372:49-53 (1995). The PTPases was shown to mediate decreasing both EGF and PDGF receptor tyrosine phosphorylation. Sorby, M. and Ostman, A., J. Biol. Chem. 271: 10963-6 (1996).
  • both PTKases and PTPases play key roles in various growth factor- or cytokine-mediated signal transduction pathways.
  • the levels of tyrosine phosphorylation required for normal cell growth and differentiation at any time are achieved through the coordinated action of PTKases and PTPases.
  • An imbalance between these enzymes results in dysfunctional signal transduction that may impairs normal cell functions leading to metabolic disorders, immune disorders, cellular transformation, and wound healing disorders.
  • the overexpression of the PTPase LAR was reported to cause a reduction in insulin receptor autophosphorylation and insulin-stimulated cellular responses. Li, P.M. et al., Cell. Signal. 8:467-73 (1996).
  • the increasing PTPase activity toward insulin receptor was reported to be a pathogenic factor in a human insulin resistance, while improving sensitivity to insulin in human adipose tissue is accompanied by decreasing of PTPase activity.
  • the imbalance between PTPase and PTKase activity was shown to play a role in impairment of insulin signal transduction under diabetes mellitus. Ahmad, F. et al., J. Clin. Invest. 100:449-58 (1997);.
  • Diabetes and systemic glucocorticoid treatment caused a severe defect in wound healing accompanied by reduced expression of PDGF receptors. Beer, H.D. et al., I Invest. Dermatol. 109:132-8(1997).
  • the levels of IGF-1 was reported to be reduced in the wound enviroment of diabetics, whereas treatment of these wounds with IGF-1 reverses diabetes-induced wound healing impairment.
  • Wound healing is impaired under ageing due to delay in appearance of growth factors such as PDGF, EGF and their receptors in the wound enviroment. Ashcroft, G.S. et al., J. Anat.
  • Hydrogen peroxide H 2 O 2 was demonstrated to be universal inhibitor of PTPase activity that inhibits specifically both cytoplasmic (PTP IB) and transmembrane (LAR) PTPases under physiological conditions by reversible oxidation of the catalytic cysteine thiolate.
  • PTP IB cytoplasmic
  • LAR transmembrane
  • H 2 O 2 was reported to inhibit LMW PTPase which downregulates PDGF and insulin receptor signal transduction. Caselli, A. et al., J. Biol. Chem. 273:32554-60 (1998).
  • EGF-induced intracellular H 2 O 2 was reported to inhibit PTPase IB, and this inhibition was found to be a fully reversible.
  • stimulation of endogenous hydrogen peroxide production is a physiologically occurring way to enhance tyrosine phosphorylation in mammalian tissues through inhibiting PTPase activity.
  • Succinic acid is the physiologically occuring substrate of succinate dehydrogenase that play a role in cellular respiration and energy metabolism.
  • succinate dehydrogenase that play a role in cellular respiration and energy metabolism.
  • This biological effect is unexpected to prior arts.
  • respiratory chain of isolated mitochondria can generate hydrogen peroxide as a minor by-product with a numerous of respiratory substrates including succinate, the inactivation of PTPases by H 2 O 2 generated in mitochondrial respiratory reactions is unknown from prior art. Boveris, A. et al., Biochem. J.
  • the present invention shows for the first time that administering to a mammal an effective amount of succinic acid or salt thereof is effective for inhibiting PTPase activity. Since inhibiting PTPase activity is effective for modulating dysfunctional signal transduction, the administering of succinic acid or salt thereof to mammals is accordingly useful in treating disease states caused by dysfunctional signal transduction.
  • Vanadates and peroxovanadates are known to be non-specific PTPases inhibitors that mimic a variety effects of insulin. Bevan, AP. et al., Mol. Cell. Biochem. 153:49-58 (1995). However, this class of compounds is toxic because each of compound contains a heavy metal atom. Moreover, evidences was presented that vanadate as well as peroxovanadate attenuates optimal mammary epithelial cell growth stimulated by combination of insulin and EGF. Mclntyre, B.S. et al., Proc. Soc. Exp. Biol. Med. 217:180-7 (1998).
  • the present invention relates to the use of succinic acid or salt thereof for inhibiting the activity of protein tyrosine phosphatases (PTPases).
  • PTPases protein tyrosine phosphatases
  • the invention is further relates to the regulation of cellular processes governed by signal transduction through the inhibition of the activity of PTPases by the succinic acid or salt thereof.
  • the invention further provides for the use of succinic acid or salt thereof in the treatment of a mammal having a disorder caused by dysfunctional signal transduction, preferably diabetes mellitus.
  • the invention further provides for the use of succinic acid or salt thereof in the treatment of a wound in a mammal.
  • the present invention provides a method of inhibiting protein tyrosine phosphatase activity, which comprises administering to a mammal in need thereof an effective amount of succinic acid or a pharmaceutically acceptable salt thereof.
  • the succinic acid or a pharmaceutically acceptable salt thereof inhibits PTPases not directly by binding to molecule of a PTPase, but through stimulating production of endogenous hydrogen peroxide which is a non-specific inhibitor both cytoplasmic and transmembrane PTPases.
  • PTPases of the invention play a role in cellular signal transduction.
  • the inhibiting PTPase activity by succinic acid or a pharmaceutically acceptable salt thereof is useful for modulating signal transduction and regulation cellular processes which are governed by signal transduction.
  • signal transduction includes the multiple pathways that are regulated by reversible phosphorylation of specific tyrosine residues on proteins involved in signaling.
  • modulating means upregulation or downregulation of a signaling pathway.
  • Cellular processes under the control of signal transduction may include, but are not limited to, normal processes such as proliferation, differentiation, transcription of specific genes, adhesion, apoptosis, and survival; and abnormal processes such as transformation, and blocking differentiation.
  • a signal may be triggered by the binding of a ligand to its receptor which transduces the signal by the phosphorylating or dephosphorylating specific tyrosine residues on proteins inside the cell.
  • proteins may include, but are not limited to, the receptor or its subunits, the receptor substrates, cytoplasmic kinases, cytoplasmic phosphatases, adapter molecules, and transcription factors.
  • receptor include, but are not limited to, insulin receptor, members of insulin-like growth factor receptor family, epidermal growth factor receptor family, transforming growth factor receptor family, fibroblast growth factor receptor family, hepatocyte growth factor receptor family, vascular endothelial growth factor receptor family, platelet-derived growth factor receptor, the T-cell receptor, the B-cell receptor, members of the type 2 to 4 cytokine receptor families, and erythropoietin receptor.
  • the adapter molecules may include the Grb proteins, IRS-1, and Zap-70.
  • the transcription factors may include nuclear factor kappa-B, and STAT proteins.
  • the ligand includes, but are not limited to, insulin, insulin-like growth factors, epidermal growth factor, platelet- derived growth factor, vascular endothelial growth factor, fibroblast growth factors, transforming growth factor, and neurotrophins, and cytokines such as growth hormone, erythropoietin, tumor necrosis factor, interleukins, and interferons.
  • the inhibiting PTPase activity by succinic acid or a pharmaceutically acceptable salt thereof can be used to upregulate signal transduction in mammals so that the effect of ligand binding to a receptor is enhanced, or mimicked if the ligand is not present.
  • the succinic acid or a pharmaceutically acceptable salt thereof exerts this effect in the signaling pathway which is normally downregulated by tyrosine dephosphorylation.
  • the means by which PTPases normally downregulate signal transduction may involve the dephosphorylation of specific phosphotyrosine residues on receptors which needed in phosphorylation of its own tyrosine residues to achieve optimal activity in the signaling pathway.
  • the inhibiting PTPase activity can be used to prevent dephosphorylation of phosphotyrosine residues on receptors which normally becomes phosphorylated upon ligand binding, thereby prolong receptor mediated signal transduction.
  • the inhibiting PTPase activity can also be used to prevent the dephosphorylation of the receptor in which the tyrosine residues become autophosphorylated due to basal activity of the receptor. In these receptors, a signal may be triggered even in the absence of ligand binding.
  • the inhibiting PTPase activity by succinic acid or a pharmaceutically acceptable salt thereof can be used to downregulate signal transduction in cells so that the effect of ligand binding to a receptor is abolished or attenuated.
  • the succinic acid or a pharmaceutically acceptable salt thereof exert this effect in the signaling pathway which normally downregulates by tyrosine dephosphorylation.
  • signaling pathways include, but are not limited to, Src family tyrosine kinases which are activated by dephosphorylation of their inhibitory site.
  • the inhibiting PTPase activity by succinic acid or a pharmaceutically acceptable salt thereof is useful in insulin-sensitive tissues for triggering or enhancing insulin receptor signal transduction by inhibiting the constitutive dephosphorylation of the phosphotyrosine sites on the activated insulin receptor. This would allow to prolong or enhance phosphorylation of the insulin receptor, thus prolong or enhance insulin receptor signal transduction.
  • the succinic acid or a pharmaceutically acceptable salt thereof can be used to trigger a signal even in absence of insulin, since insulin receptor was demonstrated to be phosphorylated at a low level even in absence of insulin. Goldstein, B.J.. J. Cell. Biochem. 48:33-42(1992).
  • the present invention provides a method of treating disease states caused by dysfunctional signal transduction, which comprises administering to a mammal in need thereof an effective amount of succinic acid or a pharmaceutically acceptable salt thereof.
  • Dysfunctional signal transduction can result from imbalance between activity of PTPases and proteine tyrosine kinases which play roles in a signaling pathway.
  • the disease states may be caused by dysfunctional signal transduction of insulin receptor, members of insulin-like growth factor receptor family, epidermal growth factor receptor family, transforming growth factor receptor family, fibroblast growth factor receptor family, hepatocyte growth factor receptor family, vascular endothelial growth factor receptor family, transforming growth factor, platelet-derived growth factor receptor, the T-cell receptor, the B-cell receptor, members of the type 2 to 4 cytokine receptor families, and erythropoietin receptor.
  • the disease states may include, but are not limited to, diabetes mellitus and its chronic complications such as nephropathy, encephalopathy, microangiopathy, retinopathy, diabetic foot ulcers caused by deficient insulin signal transduction; immune disorders caused by deficient cytokine signal transduction such as anemia and immunodeficiency; neurodegenerative disorders caused by deficient neurotrophin signal transduction; cancer; and wound healing disorders caused by deficient insulin and growth factor signal transduction under ageing, diabetes, or infections.
  • the disease state is diabetes mellitus.
  • the present invention provides a method of treating wounds, which comprises administering to a mammal in need thereof an effective amount of succinic acid or a pharmaceutically acceptable salt thereof.
  • wounds may include, but are not limited to, incised wounds, burn wounds, gastrointenstinal mucosal erosions and ulcers, periodontal wounds, and bone fractures.
  • the invention provides a method of treating disease state caused by dysfunctional signal transduction in a mammal, which comprises administering to a mammal in need thereof an effective amount of succinic acid or a pharmaceutically acceptable salt thereof stepwise or in physical combination with an effective amount of a protein tyrosine kinase activator.
  • protein tyrosine kinase activator includes ligands to receptors which contain intrinsic tyrosine kinase activity, or ligands to receptors which do not have intrinsic tyrosine kinase activity, but can stimulate tyrosine phosphorylation through association and activating cytoplasmic PTKases.
  • ligand include, but are not limited to, insulin, insulin-like growth factors, epidermal growth factor, platelet-derived growth factor, transforming growth factor, vascular endothelial growth factor, fibroblast growth factor, and neurotrophins, and cytokines such as growth hormone, erythropoietin, tumor necrosis factor, interleukins, and interferons.
  • receptor include but are not limited to, insulin receptor, members of insulin-like growth factor receptor family, epidermal growth factor receptor family, fibroblast growth factor receptor family, hepatocyte growth factor receptor family, vascular endothelial growth factor receptor family, nerve growth factor, platelet-derived growth factor, the T-cell receptor, the B-cell receptor, members of the type 2 to 4 cytokine receptor families, and erythropoietin receptor.
  • the new combination of succinic acid or a pharmaceutically acceptable salt thereof with the protein tyrosine kinase activator is synergistically effective for modulating signal transduction.
  • the combination of the invention is synergistically effective in upregulating signal transduction that normally upregulated by phosphorylation of tyrosine residues in proteins involved in the signaling pathways.
  • the succinic acid or a pharmaceutically acceptable salt thereof is administered for period of 1 day or longer, more preferably, 3 to 7 days.
  • the invention provides for the use of succinic acid or a pharmaceutically acceptable salt thereof for the manufacture of a medicament or a nutritional supplement effective for treating disease state caused by dysfunctional signal transduction in a mammal, preferably in a human.
  • Succinic acid or a pharmaceutically acceptable salt thereof is useful in nutritional and medicinal formulations according to the present invention for treating disease state caused by dysfunctional signal transduction in a mammal, preferably in a human.
  • the medicament or nutritional supplement of the invention is prepared by known procedures using well-known ingredients. In making the medicament or nutritional supplement, the active ingredients will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, and may be in the form of a capsule, tablet, paper or other container.
  • the carrier When the carrier serves as a diluent, it may be a solid, semisolid, or liquid material which acts as a vehicle, excipient, or medium for the active ingredient.
  • the nutritional supplements can be in the form of tablets, pills, powders, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules.
  • the medicament can be in the form of tablets, pills, powders, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules, aerosoles, suppositories, sterile injectable solutions, and sterile packaged powders.
  • suitable carriers include lactose, dextrose, sorbitol, mannitol, calcium phosphate, alginates, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propyl hydroxybenzoates, talc, magnesium stearate, stearic acid, and mineral oil.
  • the medicaments or nutritional supplements can additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents, or flavoring agents.
  • the orally ingestible nutritional supplement of the invention can comprise deionized water, succinic acid or salt thereof, maltodextrin, sodium caseinate, corn syrup solids, medium chain triglycerides, canola oil, calcium caseinate, soy protein, potassium citrate, magnesium chloride, sodium citrate, tricalcium phosphate, soy lecithine, sodium invention is synergistically effective in downregulating signal transduction that normally downregulated by dephosphorylation of phosphotyrosine residues in proteins involved in signaling pathways.
  • the protein tyrosine kinase activator of the composition is insulin.
  • Insulin of the invention is any standard insulin prepared by methods well-known in the art.
  • the new combination of succinic acid or a pharmaceutically acceptable salt thereof with insulin is synergistically effective in triggering or enhancing insulin receptor signal transduction that is accordingly effective for treating disease states caused by dysfunctional signal transduction of insulin receptor.
  • the disease states caused by dysfunctional insulin receptor signal transduction is diabetes mellitus.
  • Insulin is administered parenterally in the method of the invention.
  • the effective amount of insulin is selected from 2 units to 100 units per day per mammalian subject.
  • the mammal of the invention is a human.
  • the succinic acid has the chemical structure given below:
  • the pharmaceutically acceptable salt of the succinic acid is prepared by known methods from organic and inorganic bases.
  • bases include, but are not limited to, nontoxic alkali metal and akaline earth bases, for example, calcium, magnesium, lithium, sodium, and potassium hydroxide; ammonium hydroxide and nontoxic organic bases, such as triethylamine, butylamine, diethanolamine, triethanolamine, and 2-ethyl- 6-methyl-3-hydroxypyridine.
  • the salt of succinic acid can be in form of solvates with water or suitable organic solvents.
  • the succinic acid or a pharmaceutically acceptable salt thereof is preferably administered orally and topically in the methods of the invention.
  • the succinic acid or a pharmaceutically acceptable salt thereof may also be administered by a variety of other routes such as parenterally, e.g. intravenously, or subcutaneously, or intramuscularly; transdermally; or rectally.
  • the effective amount of succinic acid or a pharmaceutically acceptable salt thereof for use in the methods of the invention is selected from 0.1 milligram to 50 milligrams, more preferably from 1 mg to 20 mg per day per kilogram ascorbate, choline chloride, potassium chloride, coenzyme Q, vitamin E, molibdenium yeast, selenium yeast, carrageenan, chromium yeast, biotin, niacinamide, zink sulfate, ferrous sulfate, calcium pantothenate, vitamin A, ascorbic acid, cyancobalamine, manganese sulfate, copper gluconate, vitamine K, thiamin, pyridoxine hydrochloride, vitamine D, riboflavin, folic acid
  • Nutritional Ingredients which may also be added to nutritional supplement of the invention include fructose, soybean oil, sunflower oil, canola oil, carnitine, taurine, and other natural components.
  • the various compositions of the invention can also contain other conventional ingredients such as suspending or wetting agents, preservatives, antioxidants, thickening agents and the like.
  • the medicament of the invention in form of oral formulations can comprise a water; tetrasodium pyrophosphate or other pyrophosphate source; sodium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, or other fluoride source; alkali metal bicarbonate salt; xylitol; thickening agents such as carboxyvinyl polymers, carrageenan, hydroxyethyl cellulose, sodium carboxymethylcellulose and other water soluble salt of cellulose ethers, natural gum like gum karaya, xanthan gum, gum arabic, and gum fragacanth; colloidal magnesium aluminium silicate, lithium aluminium silicate; humectants such as glycerin, sorbitol, polyethylene glycol, propylene glycol, and other alcohols; abrasive polishing materials such as silicas, calcium carbonate, dicalcium orthophosphate dihydrate, calcium
  • the medicament of the invention in form of topical formulations may include, but are not limited to, solutions, suspensions, salves, ointments, creams, gels, aerosoles, jellies, emulsions, powders, liniments, and the like. If desired, these may be sterilized or mixed with auxiliary agents, e.g. preservatives; stabilizers such as albumine, a dissacharide, a cyclic oligosaccaride like hydroxypropyl-beta-cyclodextrin; surfactants; wetting agents; perfumes; solvents such as water; or buffers.
  • auxiliary agents e.g. preservatives
  • stabilizers such as albumine, a dissacharide, a cyclic oligosaccaride like hydroxypropyl-beta-cyclodextrin
  • surfactants e.g. preservatives
  • wetting agents e.g. preservatives
  • perfumes
  • the medicament of the invention may be in form suitable for parenteral, e.g. intravenous, subcutaneous, or intramuscular administration.
  • parenteral e.g. intravenous, subcutaneous, or intramuscular administration.
  • the succinic acid or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable carrier such as a commonly used intravenous fluid and administered by infusion.
  • a pharmaceutically acceptable carrier such as a commonly used intravenous fluid and administered by infusion.
  • physiological buffer saline or Ringer's solution can be used.
  • a sterile the active components of composition can be dissolved and administered in a pharmaceutically acceptable carrier such as pyrogen-free water (distilled) or physiological saline.
  • the medicament or nutritional supplement of the invention can be used in combination with other biologically active compounds known to the art that exhibit antidiabetic action such as peroxovanadates, and oral hypoglicemic drug like sulfonylureas and biguanides.
  • the medicament or nutritional supplement of the invention can be used in combination with other biologically active compounds known to the art that exhibit activities directed to enhancing of wound healing such as antimicrobial agents, peroxocompounds like peroxovanadates, and
  • Treating describes the managment and care of a mammal for the purpose of combating the disease, condition, or disorder and includes the administration of a compound of present invention to prevent the onset of the symptoms or complications, alleviating the symptoms or complications, or eliminating the disease, condition, or disorder.
  • EXAMPLE 1 This example shows that disodium succinate hexahydrate is effective in inhibiting PTPase activity.
  • mice Male albino Wistar rats 8-10 weeks of age 200-250 grams of body weight were used. The rats were housed at the temperature of 18 ⁇ 21°C on a 12 hour light-dark cycle. Rats were fed on a stock laboratory diet (59 % carbohydrates; 17 % protein; 3 % fat; 21 % minerals, water, cellulose) and allowed water ad libitum.
  • Adipocytes preparation Isolated rat adipocytes were prepared by the method of
  • the cells were filtered through double cheesecloth, washed three times with 5 volume of 1% BSA buffer, and resuspended in this medium.
  • the homogenate was centrifuged at 3000 g for 15 min, washed with 1% BSA buffer, and resuspended in this medium again. Then the homogenate was centrifuged at 17000 g for 30 min and washed with 1% BSA buffer. Further, prepared homogenate was centrifuged again at 30000 g for 30 min, washed with 1% BSA buffer, and rat adipocytes (4 l0 6 cells/ml) were used in the experiments.
  • Rat livers were washed with cold phosphate- buffered saline and homogenized in buffer containing 100 mM Tris-HCl, 1.15% KCl, and pH 7.4. The homogenate was centrifuged at 200 g for 10 min to remove debris, and a postnuclear supernatant (35.6 mg of protein per ml) was used in the experiments.
  • Hydrogen peroxide assay To determine hydrogen peroxide production in the rat tissue fractions, the assay based on the horseradish peroxidase-mediated oxidation of phenol red by H 2 O 2 was used. Pick, E. and Keisari, Y., J. Immunol. Methods 38: 161- 70(1980). The rat adipocytes or the liver supernatant were incubated at 37°C in a buffer containing 50 mM Tris-HCl, 5 mM EDTN 0.02% digitonin, pH 7.6, and with or without (control) 2 mM disodium succinate hexahydrate. An aliquot of the cellular fraction was used to estimate H 2 O 2 release at 0, 5, 10 and 15 min of the incubation. Results are presented in Tables 1 to 2 as mean ⁇ S.E.M., and as H2O2 concentration in % of control.
  • disodium succinate stimulates production of endogenous hydrogen peroxide which is a well-known inhibitor of PTPase activity.
  • Disodium succinate hexahydrate was assayed for its activity in inhibiting PTPase activity in the cellular fractions (rat adipocytes and liver supernatant).
  • the PTPase assay 60 ⁇ l of the cell fraction was incubated in 40 ⁇ l of a reaction buffer containing 50 mM Tris/HCl, pH 7.5 and 10 mM phosphotyrosine as the substrate, and with or without (control) 2 mM disodium succinate hexahydrate. After a 5, 10, or 20 min incubation period at 37°C, the reaction was terminated by the addition of 40 ⁇ l 10% BSA and 120 ⁇ l 25% tricloroacetic acid.
  • the probes were centrifuged at 6000 g for 10 min and 20 ⁇ l of the supernatant were mixed with 380 ⁇ l of 50 mM Tris-HCl, pH 7.5 and 100 ⁇ l of the malachite green colour reagent (Baykov, A. A. et al. Anal. Biochem. 171:266-70 (1988). After a 10 min incubation period at a room temperature, the extinction was measured at 655 nm in microtiter plates. The specific PTPase activity was calculated by using a standard graph ranging inorganic phosphate (Pi). Results are presented in Tables 3 to 4 as mean ⁇ S.E.M., and as PTPase activity in % of control.
  • the administering of disodium salt of succinic acid is effective in inhibiting PTPase activity.
  • EXAMPLE 2 This example shows that administering disodium succinate mimics insulin in totally pancreatectomized rats.
  • Plasma free fatty acids levels were determined by enzymatic method with a commercially available kit (Waho Chemicals Gmbh, Neuss, Germany) with Multistat 3 centrifugal analyzer (Instrumentation Laboratories, Lexington, USA).
  • Serum glucose concentrations were determined by the glucose oxidase method with a kit (Lachema, Slov.) with glucose analyzer (Beckman, Fullerton, Calif, USA).
  • Plasma insulin concentrations were determined by a double-antibody radioimmunoassay kit (Kabi Pharmacia Diagnostics, Uppsala, Sweden) using a rat insulin standard (Novo Research Institute, Bagsvard, Denmark).
  • Plasma C-peptide concentrations were determined with a kit (Dako, Denmark) using a rat C-peptide standard.
  • Plasma triglycerides and cholesterol concentrations in High Density Lipoprotein (HDL) and Low Density Lipoprotein (LDL) were determined by enzymatic methods with kits (Boeringher Mannheim, Mannheim, Germany) with Multistat 3 F LS apparatus (Instrumentation Laboratories, Lexington, USA).
  • the male Wistar rats maintained as described in example 1 of the invention were used in these experiments.
  • the rats were anaesthetized with 1% solution of nembutal (0.2 ml/100 g of body weight, intramuscularly), and the total pancreatectomy (TP) was made.
  • the vessels were ligated, the abdominal cavities were purified and then sewed.
  • the values of plasma insulin and C-peptide levels in totally pancreatectomized rats treated by the succinate were below the detection limits of the used assays during the 60 days of the experiment.
  • the detection limit was 3 pmol 1 for plasma insulin assay, and 0.0165 nmol/1 for plasma C-peptide assay.
  • Fasting plasma insulin levels, plasma C-peptide levels, serum glucose levels, plasma free fatty acid levels (FFA), plasma cholesterol levels, plasma triglycerides levels, plasma high density lipoprotein cholesterol levels (HDL), and plasma low density lipoprotein cholesterol levels (HDL) were measured in rats by tail clipping method before total pancreatectomy (TP), and after TP.
  • the data are presented in Tables 5 through 7 as mean ⁇ S.E.M. Table 5.
  • Mean fasting plasma Insulin. C-peptide. and serum Glucose levels in totally pancreatectomized rats (n 10) treated by disodium succinate.
  • Treatment period 10 0.51 ⁇ 0.08 0.92 ⁇ 0.14 1.54 ⁇ 0.34 20 0.51 ⁇ 0.04 1.46 ⁇ 0.22 1.52 ⁇ 0.41 40 0.52 ⁇ 0.06 1.27 ⁇ 0.34 1.66 ⁇ 0.19 60 0.59 ⁇ 0.04 1.30 ⁇ 0.38 1.51 ⁇ 0.27
  • the administering disodium succinate to the totally pancreatectomized rats results in 4- fold decreasing in the pathologically elevated glucose levels even in the absence (non- detectable quantities) of insulin in the rat plasma.
  • the succinic acid salt exhibits insulinomimetic action even in the absence of insulin. Since pancreatectomy in rats is animal model of diabetes, the administering the succinic acid salt is effective therapy for treating diabetes mellitus.
  • This example shows that administering succinic acid and insulin is synergistically effective in treating diabetes mellitus.
  • NIDDM Non-insulin Dependent Diabetes Mellitus
  • NIDDM humans were metabolically stable and had fasting glucose levels more than
  • NIDDM human patients is effective for treating diabetes mellitus.
  • This example shows that administering an effective amount of disodium salt of succinic acid is effective for treating wounds.
  • a suppurative wounds were prepared by the method of Eliseev. Eliseev, N.G. "Connective tissue", Moscow (1961).
  • the 30 male Wistar rats maintained as described in example 1 of the invention were used in this experiment.
  • the rats were anaesthetized with 1% solution of nembutal (0.2 ml/100 g of body weight, intramuscularly). Incisions of 15 to 20 mm length in inguinal region of rats were made.
  • a four celluloid balls of 2 mm diameter were entered into the each wound.
  • the topical treating of suppurative wounds by disodium salt of succinic acid significantly enhances wound healing.
  • the rate of healing was found to be more than 2- fold higher in the group of rats treated by the succinate in comparison with the control untreated rats for all consecutive phases of wound healing, including the complete re- epitalization, an ultimate goal in wound healing.
  • administering the succinic acid salts is effective in treating wounds.

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Abstract

Cette invention concerne l'utilisation d'acide succinique, ou de sels d'acide succinique pharmaceutiquement acceptables, dans la fabrication de médicaments ou d'additifs nutritionnels efficaces qui permettent de traiter des états maladifs induits par un dysfonctionnement de la transduction de signaux, de préférence le diabète, et qui permettent de cicatriser des blessures. Cette invention concerne plus précisément des procédés qui permettent d'inhiber l'activité de la protéine tyrosine phosphatase, et de traiter des états maladifs induits par un dysfonctionnement de la transduction de signaux. Ces procédés consistent à administrer de l'acide succinique, ou des sels pharmaceutiquement acceptable de celui-ci, soit seul, soit en combinaison avec des activateurs de protéine tyrosine kinases, de préférence avec de l'insuline.
EP99933303A 1999-04-05 1999-04-05 Utilisation d'acide succinique ou de ses sels dans le traitement du diabete et la cicatrisation de blessures Withdrawn EP1165063A1 (fr)

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AU2000263274A1 (en) * 2000-04-10 2001-10-23 Olga Evgenievna Kolesova Synergistic compositions containing choline base and succinic acid for insulin resistance and diabetes
US6797693B2 (en) 2001-06-07 2004-09-28 Wyeth Methods using PTPase inhibitors and insulin
AUPR626001A0 (en) * 2001-07-10 2001-08-02 Mcgregor, Neil A method of treatment and/or prophylaxis
AU2002344698B2 (en) * 2001-07-10 2006-07-27 Penam Investments Pty Ltd A method of treatment and/or prophylaxis of ulcers
WO2003055476A1 (fr) * 2001-12-25 2003-07-10 Pomytkin Igor A Utilisation d'acide succinique et de ses sels dans l'inhibition de l'agregation plaquetaire
AU2003236156A1 (en) * 2003-04-24 2005-01-04 Shin-Jen Shiao Pharmaceutical compositions used for immune disease treatment and improvement
EP1682570A4 (fr) * 2003-10-27 2006-12-06 Medvet Science Pty Ltd Motif de liaison et methodes de regulation d'une fonction cellulaire
JP5110898B2 (ja) * 2007-02-16 2012-12-26 キヤノン株式会社 吐出用液体及び吐出方法
AU2019291662B2 (en) 2018-06-22 2025-04-17 Enzene Biosciences Limited Succinic acid and derivatives for the treatment of haemotological disorders

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JPS56125312A (en) * 1980-03-09 1981-10-01 Lion Corp Remedy for damage
JPS61171417A (ja) * 1985-01-23 1986-08-02 Wakunaga Seiyaku Kk 抗糖尿病剤
JP2844103B2 (ja) * 1990-02-09 1999-01-06 株式会社コーセー 皮膚外用剤
JP3581157B2 (ja) * 1992-08-19 2004-10-27 株式会社ミツカングループ本社 低耐糖能者用食品
JP3854326B2 (ja) * 1995-10-27 2006-12-06 株式会社ノエビア コラゲナーゼ阻害剤
ATE208208T1 (de) * 1996-06-20 2001-11-15 Novo Nordisk As Halogenid-enthaltende insulinzubereitungen

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