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US-12622887-B2 - Liquid pharmaceutical compositions comprising citrate and carnitine

US12622887B2US 12622887 B2US12622887 B2US 12622887B2US-12622887-B2

Abstract

A composition comprising citrate and carnitine which activates the production of the protein Klotho by renal tubule cells protecting them, thus reducing the oxidative stress conditions thereof, which can be used for the conservation, perfusion and reperfusion of organs for organ transplants is disclosed. Furthermore, the composition in the liquid and dietary/nutraceutical formulation can be used for reducing the nephrotoxicity induced by iodinated contrast means, antibiotics, NSAIDs 10 or agents that induce oxidative stress.

Inventors

  • Giuseppe CASTELLANO
  • Rossana FRANZIN
  • Alessandra STASI
  • Fabio Sallustio
  • Arduino Arduini

Assignees

  • IPERBOREAL PHARMA SRL
  • Giuseppe CASTELLANO
  • PUERAPULIAE INVEST SRL

Dates

Publication Date
20260512
Application Date
20200514
Priority Date
20190529

Claims (16)

  1. 1 . A liquid pharmaceutical composition comprising: (a) active ingredients citrate and carnitine; and (b) a pharmacologically acceptable excipient, wherein the citrate is at a concentration of about 5 mM in the liquid, and the carnitine is at a concentration of between 2.5 mM and 5 mM in the liquid, wherein the citrate and carnitine are formulated with: (i) sodium bicarbonate, potassium phosphate monobasic, potassium phosphate dibasic trihydrate and potassium chloride; (ii) potassium lactobionate, KH 2 PO 4 , MgSO 4 , raffinose, adenosine, glutathione, allopurinol and hydroxyethyl starch; (iii) sodium, potassium, magnesium, calcium, ketoglutarate/glutamic acid, histidine, mannitol and tryptophan; or (iv) glutathione, mannitol, lactobionic acid, glutamic acid, sodium hydroxide, calcium chloride dihydrate, potassium chloride, magnesium chloride hexahydrate and histidine.
  2. 2 . The liquid pharmaceutical composition according to claim 1 wherein the carnitine is at a concentration of about 5 mM in the liquid.
  3. 3 . The liquid pharmaceutical composition according to claim 1 wherein the citrate is at a concentration of 5 mM in the liquid, and the carnitine in a concentration of 5 mM in the liquid.
  4. 4 . The liquid pharmaceutical composition according to claim 1 further comprising at least one polyol.
  5. 5 . The liquid pharmaceutical composition according to claim 4 wherein the polyol is selected in the group consisting of Erythritol, Sorbitol, Xylitol, Mannitol, Maltitol, Isomalt, Lactitol, and polyglycidol.
  6. 6 . The liquid pharmaceutical composition according to claim 1 , wherein the citrate comprises a sodium citrate.
  7. 7 . The liquid pharmaceutical composition according to claim 1 formulated in as a physiological saline solution.
  8. 8 . The liquid pharmaceutical composition according to claim 7 , wherein the physiological saline solution further comprises an albumin.
  9. 9 . The liquid pharmaceutical composition of claim 7 , formulated with a preservation solution, a conservation solution, a perfusion solution and/or a reperfusion solution.
  10. 10 . The liquid pharmaceutical composition of claim 9 , wherein the preservation solution, conservation solution, perfusion solution and/or reperfusion solution comprises a solution selected from the group consisting of: Euro-Collins solution, University of Wisconsin solution, histidine-tryptophan-ketoglutarate solution, Marshall solution, and Institute Georges Lopez solution.
  11. 11 . The liquid pharmaceutical composition of claim 7 , formulated for use as a preservation solution and/or a conservation solution for an organ transplant.
  12. 12 . The liquid pharmaceutical composition of claim 7 , formulated as a perfusion solution and/or a reperfusion solution for an organ transplant.
  13. 13 . The liquid pharmaceutical composition of claim 1 , wherein the citrate and carnitine are formulated with a combination of compounds consisting of: sodium bicarbonate, potassium phosphate monobasic, potassium phosphate dibasic trihydrate and potassium chloride.
  14. 14 . The liquid pharmaceutical composition of claim 1 , wherein the citrate and carnitine are formulated with a combination of compounds consisting of: potassium lactobionate, KH 2 PO 4 , MgSO 4 , raffinose, adenosine, glutathione, allopurinol and hydroxyethyl starch.
  15. 15 . The liquid pharmaceutical composition of claim 1 , wherein the citrate and carnitine are formulated with a combination of compounds consisting of: sodium, potassium, magnesium, calcium, ketoglutarate/glutamic acid, histidine, mannitol and tryptophan.
  16. 16 . The liquid pharmaceutical composition of claim 1 , wherein the citrate and carnitine are formulated with a combination of compounds consisting of: glutathione, mannitol, lactobionic acid, glutamic acid, sodium hydroxide, calcium chloride dihydrate, potassium chloride, magnesium chloride hexahydrate and histidine.

Description

FIELD OF THE INVENTION The present invention relates to the sector of pharmaceuticals and more in particular a composition comprising citrate and carnitine which is able to activate the production of the protein Klotho by the renal cells and therefore is able to reduce damage due to oxidative stress because of exogenous and endogenous causes which can affect the kidneys. The formulation can be used to reduce the nephrotoxicity induced by iodinated contrast media, antibiotics or NSAIDs and by agents that induce oxidative stress. Within the context of the organ transplant sector, the composition can be effectively used for the preservation, perfusion and reperfusion of organs for transplant and for preventing early renal ageing. BACKGROUND Klotho protein exists in three different isoforms: α, β e γ. In humans, Klotho α is codified by the gene KL and codifies for an enzyme belonging to the beta-glucosidase family mainly expressed in the distal tubule of the kidney but also at the choroid plexus level of the encephalus, in the parathyroid glands, in the bladder, in the skeletal muscle, in the placenta, in the thyroid gland and in the endothelial cells of the aorta and of the renal artery. (Donate-Correa J, Expression of FGF23/KLOTHO system in human vascular tissue. Int J Cardiol. 2013; Lim K, Vascular Klotho deficiency potentiates the development of human artery calcification and mediates resistance to fibroblast growth factor 23. Circulation. 2012, Lim K, α-Klotho Expression in Human Tissues. J Clin Endocrinol Metab. 2015). Klotho α, simply known as Klotho, is a protein that contrasts ageing and inflammation; its soluble form mainly released by the kidney is secreted into the blood and performs a protective function throughout the whole organism, in particular at cardiovascular and central nervous system level. In particular, the soluble form α Klotho reduces calcifications in the arteries, protects the heart from hypertrophy (Xie J, Cardioprotection by Klotho through downregulation of TRPC6 channels in the mouse heart. Nat Commun. 2012) through the inhibition of endothelial dysfunction and oxidative stress on the smooth muscle cells (Mencke R, Hillebrands J L; The role of the anti-ageing protein Klotho in vascular physiology and pathophysiology. Ageing Res Rev. 2017)(Donate-Correa J, Klotho in cardiovascular disease: Current and future perspectives. World J Biol Chem. 2015). At central nervous system level, α Klotho plays an essential role in promoting cognitive abilities and in contrasting neurological and psychiatric disorders (Vo H T., Klotho, the Key to Healthy Brain Aging? Brain Plast. 2018). In animal models, the increased expression of this protein has displayed a cognitive improvement and greater resistance to Alzheimer-related neurodegeneration. Furthermore, in in vitro cell cultures, it has been observed that the protein Klotho induces greater resistance to oxidative stress and significant protection from cytotoxicity induced by protein A13 and by glutamate associated with Alzheimer's disease. (Cararo-Lopes M M, The relevance of α-KLOTHO to the central nervous system: Some key questions. Ageing Res Rev. 2017) The trans-membrane form of α Klotho which acts as a co-receptor of FGF23 (Fibroblast Growth Factor-23), is constituted of 130 kDa, whereas the secreted forms that can be detected in serum, urine and cerebrospinal fluid can form both due to cleavage of the transmembrane protein and due to alternative splicing (forming the isoform of 70 kDa). In the renal tubule, the membrane protein Klotho regulates the reabsorption of phosphate and controls the metabolism of vitamin D, involved in the homeostasis of calcium. The secreted forms, instead, act independently from the FGF23 receptor and are diffused with hormonal action along the blood stream, regulating the transduction of the insulin signal (Schmid C., Growth hormone and Klotho. J Endocrinol. 2013) and the transduction route of Wnt with anti-oxidizing and anti-ageing effect. Transgenic mice without Klotho develop a syndrome similar to accelerated ageing, with premature death, osteoporosis, blindness, arteriosclerosis and vascular calcifications, in particular they display a defect in endothelium-dependent vasodilation and in angiogenesis. The protection by the protein Klotho of the cardiovascular system takes place by means of controlling the release of nitric oxide, a known anti-oxidant molecule that contrasts vasoconstriction and endothelial dysfunction. Unlike knock-out mice, mice which hyper-express Klotho live longer than normal mice. In relation to β Klotho, it is known that it controls the metabolism of lipids, the homeostasis of glucose and the release of bile (Xu Y. Molecular basis of Klotho: from gene to function in aging. Endocr Rev. 2015; Yamamoto M, et al. Regulation of oxidative stress by the anti-aging hormone klotho. J Biol Chem 2005; Ito S., Impaired negative feedback suppression of bile acid synthesis in mice lacking beta Klotho.