Pharmaceutical composition for intravenous injection

Abstract

The primary purpose of the present invention is to provide, for albumin deficiency symptoms which requires administration of zinc, a formulation that is capable of further improving the efficiency of zinc replenishment. The secondary purpose of the present invention is to provide a formulation that has a treatment effect for iron overload disorders.　In the present invention, a first pharmaceutical composition, for intravenous injection and containing zinc ions and serum albumin, is capable of more efficiently replenishing zinc for albumin deficiency symptoms that requires administration of zinc, and a second composition, for intravenous injection and containing zinc ions, is capable of removing iron in vivo for iron overload disorders.

Description

Intravenous pharmaceutical composition

[0001] The present invention relates to an intravenous pharmaceutical composition that efficiently supplies zinc for treating the concomitant symptoms of hypoalbuminemia and hypozincemia, and an intravenous pharmaceutical composition that has an in vivo iron removal effect for treating iron overload.

[0002] Zinc plays an important role in many physiological functions and is one of the important nutrients for humans. Zinc deficiency reduces protein synthesis in the body, making organs and cells where protein synthesis is active more susceptible to damage. For example, zinc deficiency is known to increase the risk of developing taste disorders, dermatitis, hair loss, anemia, stomatitis, male sexual dysfunction, susceptibility to infection, and osteoporosis. Furthermore, zinc enzymes play an important role in nitrogen and ammonia metabolism in the liver.

[0003] Several factors causing zinc deficiency have been reported. For example, Non-Patent Document 1 reports that the amount of zinc accumulated in hepatocytes decreases by 70% due to the lack of albumin, Non-Patent Document 2 reports that albumin is involved as a factor related to hypozincemia in dialysis patients, Non-Patent Document 3 reports that albumin is involved in the blood zinc concentration in patients with liver cirrhosis, and Non-Patent Document 4 speculates that a decrease in albumin due to liver disease makes zinc more likely to bind to amino acids, resulting in increased urinary excretion of zinc and a decrease in its blood concentration.

[0004] Furthermore, Non-Patent Document 5 describes the administration of zinc acetate to cirrhosis patients with zinc deficiency, and Non-Patent Document 6 describes the administration of zinc acetate to cirrhosis patients with hyperammonia and hypozincemia. As described in Non-Patent Document 5, the therapeutic effect of zinc administration on hyperammonemia has been reported in a small number of cirrhosis patients, while a systematic review in Non-Patent Document 7 reports that serum ammonia in cirrhosis patients does not improve with the administration of zinc acetate or zinc sulfate.

[0005] Iron, like zinc, is an essential trace element. Iron itself, iron as a component of heme, and iron-sulfur complexes are important nutrients for maintaining homeostasis. While the physiological lack of an iron excretion pathway in the body is rational for ensuring iron as an essential trace element, it also suggests that the body is prone to iron overload. Iron circulates in the body bound to the glycoprotein transferrin. However, in pathological conditions of iron overload, increased levels of non-transferrin-bound iron (NTBI) have been reported to catalyze the Fenton reaction, generating hydroxyl radicals and thus playing a key role in the pathogenesis of various diseases. Clinically, iron overload can be caused by mutations in iron-related genes, continuous blood transfusions, and intravenous iron administration.

[0006] Iron overload is treated with iron chelation therapy. Deferoxamine was previously used as an iron chelator, but its short half-life required continuous injections to achieve efficacy, making it difficult to administer to outpatients. Although deferasirox, an oral agent with a longer half-life, is now available, these iron chelators are associated with specific adverse events, such as visual impairment, hearing impairment, renal damage, liver damage, and gastrointestinal symptoms. Non-Patent Document 8 reports that, in addition to ferrokinetic markers that indicate therapeutic efficacy, these adverse events must also be monitored regularly.

[0007] Basic findings regarding the in vivo interaction between iron and zinc and the bioabsorption of both elements have been reported in Non-Patent Document 9, which reports that Zip 14, a transporter present in the cell membrane, plays a role in zinc and iron metabolism by mediating the uptake of zinc and NTBI into cells, and that iron uptake by Zip 14 is inhibited by zinc. Clinical findings supporting this absorption antagonism include the association between hypozincemia and iron overload, specifically, the inverse correlation between serum zinc concentration and serum ferritin concentration, which reflects tissue iron storage, in patients with chronic liver disease, and zinc deficiency promoting iron overload and ultimately inducing fatty liver through lipid peroxidation.

[0008] Zinc uptake by isolated rat liver parenchymal cells, Biochimica et Ciophysica Acta, 538 (1978) 435-444. Actual state of serum zinc concentration distribution in dialysis patients - Factors related to hypozincemia -, Journal of Dialysis 51 (6): 369-377, 2018. Significance of serum zinc concentration measurement in treatment strategies for liver cirrhosis, Liver, Vol. 62, suppl. (1) 2021 Mini-Oral, 57th Annual Meeting of the Japan Society of Hepatology, SY4-8. Ammonia metabolism and hepatic encephalopathy. Hepatology research, 2004, 30: 73-80. Recent findings on zinc deficiency and chronic liver disease - Focusing on our own cases -, Zinc nutrition therapy, Vol. 10, No. 2, 2020, 54-63. Effect of zinc on liver cirrhosis with hyperammonemia: A preliminary randomized, placebo-controlled double-blind trial, Nutrition 30 (2014) 1409-1414. Zinc supplementation in patients with cirrhosis and hepatic encephalopathy: a systematic review and meta-analysis, Shen et al. Nutrition Journal (2019) 18:34. Post-transfusion iron overload: A reference guide for clinical management (Reiwa 1 revised edition). Zip14 (Slc39a14) mediates non-transferrin-bound iron uptake into cells. Juan P. Liuzzi, Fikret Aydemir, Hyeyoung Nam, Mitchell D. Knutson, and Robert J.CousinsExacerbation of Insulin Resistance and Hepatic Steatosis Deriving from Zinc Deficiency in Patients with HCV-Related Chronic Liver Disease Takashi Himoto.

[0009] The liver plays a central role in regulating short-term zinc homeostasis and storing zinc. For symptoms of insufficient zinc accumulation in the liver, intravenous injection of zinc salts is expected to increase serum zinc concentration, and normalize serum zinc levels, thereby increasing liver zinc. For example, for symptoms of both albumin deficiency and zinc deficiency, administering zinc preparations as described in Non-Patent Document 4 can be expected to have a certain therapeutic effect, but the effect is limited, and there is still room for improvement. In addition, for the treatment of hyperammonemia in patients with liver cirrhosis accompanied by albumin deficiency, administering zinc preparations as described in Patent Document 5 and Patent Document 6 can be expected to have a certain therapeutic effect, but the effect is limited, and there is still room for improvement.

[0010] Furthermore, drug treatment for iron overload requires continuous injection and is accompanied by adverse events as described in Non-Patent Document 7. Therefore, physical constraints during administration and administration restrictions for patients with specific backgrounds are cited as issues.

[0011] Therefore, a first object of the present invention is to provide a pharmaceutical formulation that can further improve the efficiency of zinc supplementation for albumin deficiency symptoms that require zinc administration.

[0012] A second object of the present invention is to provide a novel pharmaceutical formulation that has an in vivo iron removal effect for iron overload syndrome, does not use iron chelating agents that are associated with specific adverse events, and can be used easily.

[0013] As a result of extensive research, the present inventors have unexpectedly discovered that, first, the efficiency of zinc supplementation is improved by intravenous administration of a new pharmaceutical composition containing both zinc and serum albumin, and second, that intravenous administration of zinc provides an in vivo iron removal effect comparable to that of known methods for iron overload.The present invention was completed through further research based on these findings.

[0014] That is, the present invention provides the following aspects (a first intravenous pharmaceutical composition according to Items 1 to 11, and a second intravenous pharmaceutical composition according to Items 12 to 23). Item 1. An intravenous pharmaceutical composition comprising zinc ions and serum albumin. Item 2. The intravenous pharmaceutical composition according to Item 1, wherein the zinc ions and the serum albumin form a conjugate. Item 3. The intravenous pharmaceutical composition according to Item 1 or 2, wherein the zinc ion source is a water-soluble zinc salt. Item 4. The intravenous pharmaceutical composition according to Item 3, wherein the water-soluble zinc salt is zinc sulfate. Item 5. The intravenous pharmaceutical composition according to any one of Items 1 to 4, wherein the content of serum albumin per mole of zinc ions is 0.35 mol or more. Item 6. The intravenous pharmaceutical composition according to any one of Items 1 to 5, wherein the content of zinc ions at the time of use is 0.1 mM or more. Item 7. The intravenous pharmaceutical composition according to any one of Items 1 to 6, which does not contain carbohydrates. Item 8. Item 9. The intravenous pharmaceutical composition according to any one of Items 1 to 7, which is an injection. Item 10. The intravenous pharmaceutical composition according to any one of Items 1 to 8, which is used to treat hypoalbuminemia, a condition requiring zinc administration. Item 11. The intravenous pharmaceutical composition according to Item 9 or 10, which is used to treat hyperammonemia. Item 12. The intravenous pharmaceutical composition according to Item 12, which contains zinc ions and is used to treat iron overload. Item 13. The intravenous pharmaceutical composition according to Item 12, wherein the zinc ion source is a water-soluble zinc salt. Item 14. The intravenous pharmaceutical composition according to Item 13, wherein the water-soluble zinc salt is zinc sulfate. Item 15. The intravenous pharmaceutical composition according to any one of Items 12 to 14, wherein the zinc ion content at the time of use is 0.1 mM or more. Item 16. The intravenous pharmaceutical composition according to any one of Items 12 to 15, which is an injection. Item 17. Item 17. The pharmaceutical composition for intravenous injection according to any one of Items 12 to 16, further comprising serum albumin. Item 18. The pharmaceutical composition for intravenous injection according to Item 17, wherein the zinc ion and the serum albumin are conjugated. Item 19. The pharmaceutical composition for intravenous injection according to Item 17 or 18, wherein the content of the serum albumin per mole of the zinc ion is 0.35 moles or more.Item 20. The intravenous pharmaceutical composition according to any one of Items 17 to 19, wherein the iron overload is accompanied by hypoalbuminemia. Item 21. The intravenous pharmaceutical composition according to Item 20, wherein the hypoalbuminemia is hypoalbuminemia-hypozincemia. Item 22. The intravenous pharmaceutical composition according to any one of Items 12 to 21, wherein the iron overload is accompanied by hyperammonemia. Item 23. The intravenous pharmaceutical composition according to any one of Items 12 to 22, which does not contain carbohydrates. Item 24. A method for supplementing zinc, comprising the step of intravenously injecting a pharmaceutical composition containing a therapeutically effective amount of zinc ions and serum albumin to a subject suffering from hypoalbuminemia and in need of zinc administration. Item 25. Use of the intravenous pharmaceutical composition containing zinc ions and serum albumin for the manufacture of a zinc supplement for hypoalbuminemia-hypozincemia. Item 26. An intravenous pharmaceutical composition containing zinc ions and serum albumin for use in supplementing zinc in hypoalbuminemia-hypozincemia. Item 27. Item 27. A method for treating iron overload, comprising intravenously injecting an effective amount of zinc ions into a subject in need of such treatment. Item 28. Use of zinc ions for the manufacture of a pharmaceutical composition for intravenous administration for treating iron overload. Item 29. Zinc ions for use in treating iron overload.

[0015] According to the present invention, first, a pharmaceutical formulation is provided that can further improve the efficiency of zinc supplementation for symptoms such as hypoalbuminemia that require zinc administration, and second, a pharmaceutical formulation is provided that has an in vivo iron removal effect for iron overload syndrome.

[0016] 1 shows the concentration of zinc ions bound to albumin in intravenous solutions containing 1 mM zinc sulfate and different concentrations of albumin (Test Example 1-1). 1 shows the results of comparing the binding strength of zinc ions to albumin between an intravenous solution containing zinc sulfate and albumin and an intravenous solution containing zinc acetate or zinc chloride and albumin (Test Example 1-2). 1 shows the serum zinc concentration after a single administration of the first intravenous pharmaceutical composition (Zn-Alb) of the present invention to a rat model of hypoalbuminemia. 1 shows the increase in serum zinc after a single administration of the first intravenous pharmaceutical composition (Zn-Alb) of the present invention to a rat model of hypoalbuminemia. 1 shows the serum zinc concentration after repeated administration of the first intravenous pharmaceutical composition (Zn-Alb) of the present invention to a rat model of hypoalbuminemia. 1 shows the increase in serum zinc when the first intravenous pharmaceutical composition of the present invention (Zn-Alb) is repeatedly administered to a rat model of hypoalbuminemic hypozincemia. 2 shows the serum ammonia-reducing effect when the first intravenous pharmaceutical composition of the present invention (Zn-Alb 200, 400, 600) is repeatedly administered to a rat model of hypoalbuminemic hypozincemia accompanied by hyperammonemia. 3 shows the liver zinc-increasing effect when the first intravenous pharmaceutical composition of the present invention (Zn-Alb 200, 400, 600) is repeatedly administered to a rat model of hypoalbuminemic hypozincemia. 4 shows a comparison of the plasma ferritin-reducing effect between the second intravenous pharmaceutical composition of the present invention (Zn, Zn-Alb) and deferasirox (a therapeutic drug for iron overload). 5 shows a comparison of the plasma iron-reducing effect between the second intravenous pharmaceutical composition of the present invention (Zn, Zn-Alb) and deferasirox (a therapeutic drug for iron overload). 1 shows a comparison of the unsaturated iron-binding capacity (UIBC) increasing effect of the second intravenous pharmaceutical composition of the present invention (Zn, Zn-Alb) and deferasirox (a drug for treating iron overload). 2 shows the serum iron-reducing effect when the second intravenous pharmaceutical composition of the present invention (Zn-Sul, Zn-Alb) is repeatedly administered to an iron overload model rat. 3 shows the serum iron-reducing effect when the second intravenous pharmaceutical composition of the present invention (Zn and Zn-Alb 200,600) is repeatedly administered to an iron overload model rat accompanied by hypoalbuminemia and hypozincemia.

[0017] 1. First Intravenous Pharmaceutical Composition The first intravenous pharmaceutical composition of the present invention is characterized by containing zinc ions and serum albumin. The first intravenous pharmaceutical composition of the present invention will be described in detail below.

[0018] Zinc Ion Source A water-soluble zinc salt can be used as the zinc ion source. The water-soluble zinc salt is not particularly limited as long as it is a pharmaceutically acceptable salt. Examples of water-soluble zinc salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, and sulfuric acid; and salts of organic acids such as acetic acid, citric acid, tartaric acid, lactic acid, succinic acid, fumaric acid, maleic acid, and methanesulfonic acid. The water-soluble zinc salt may be an anhydride or a hydrate.

[0019] In the first pharmaceutical composition for intravenous injection of the present invention, the water-soluble zinc salt may be one of the above salts alone or a combination of two or more thereof. Among the above salts, from the viewpoint of further improving the efficiency of zinc supplementation, preferred are salts of inorganic acid salts, more preferred are sulfate salts (i.e., zinc sulfate), and even more preferred is zinc sulfate heptahydrate.

[0020] The zinc ion content may be appropriately determined depending on the desired degree of improvement in zinc replenishment efficiency and / or the need for dilution or dissolution at the time of use. For example, the zinc ion concentration in the intravenous pharmaceutical composition at the time of use may be, for example, 0.1 mM or more, preferably 0.5 mM or more, more preferably 0.7 mM or more, even more preferably 0.8 mM or more, and even more preferably 0.9 mM or more. The upper limit of the zinc ion concentration is not particularly limited, but may be, for example, 5 mM or less, preferably 4 mM or less, more preferably 3 mM or less, even more preferably 2 mM or less, even more preferably 1.5 mM or less, and even more preferably 1.2 mM or less. Specific ranges of the zinc ion concentration in the intravenous pharmaceutical composition at the time of use include, for example, 0.1 to 5 mM or 0.1 to 4 mM, preferably 0.5 to 3 mM, more preferably 0.7 to 2 mM, even more preferably 0.8 to 1.5 mM, and even more preferably 0.9 to 1.2 mM.

[0021] Serum Albumin The origin of serum albumin is not particularly limited, as long as it is the same as that of the animal species to which the pharmaceutical composition for intravenous injection is to be administered.

[0022] The content of serum albumin may be appropriately determined depending on the desired degree of improvement in zinc supplementation efficiency, and may be, for example, 0.35 mol or more, preferably 0.4 mol or more, more preferably 0.5 mol or more, or 0.55 mol or more, relative to 1 mol of zinc ion. The upper limit of the content of serum albumin relative to 1 mol of zinc ion is not particularly limited, and may be, for example, 1 mol or less, preferably 0.9 mol or less, more preferably 0.8 mol or less, and even more preferably 0.7 mol or less, or 0.65 mol or less. Specific ranges of the content of serum albumin relative to 1 mol of zinc ion include, for example, 0.35 to 1 mol or 0.35 to 0.9 mol, preferably 0.4 to 0.8 mol, more preferably 0.5 to 0.7 mol or 0.55 to 0.65 mol.

[0023] Forms of Zinc Ions and Serum Albumin In the first pharmaceutical composition for intravenous injection of the present invention, zinc ions and serum albumin form a conjugate. Specifically, some or all of the zinc ions can form a conjugate with serum albumin. In the present invention, the term "zinc ions" refers to both free zinc ions and zinc ions conjugated with serum albumin. The amount of zinc ions refers to the total amount of free zinc ions and zinc ions conjugated with serum albumin. When the serum albumin is human serum albumin, the conjugate is considered to be an embodiment in which zinc ions are bound between His67 and Asn99 in domain I and His247 and Asp249 in domain II of human serum albumin.

[0024]

[0039] In addition to the above-mentioned components, the first pharmaceutical composition for intravenous injection of the present invention may contain other base materials and / or additives that can be used in pharmaceutical compositions for intravenous injection. Such base materials typically include water, and additives include isotonic agents (typically sodium chloride) and pH adjusters.

[0025] The first intravenous pharmaceutical composition of the present invention preferably does not contain any nutrients that can be used in intravenous pharmaceutical compositions in addition to the above-mentioned components, but such nutrients include carbohydrates (glucose, fructose, xylitol, etc.), amino acids, vitamins, etc.

[0026] The first intravenous pharmaceutical composition of the present invention may contain, in addition to the above-mentioned components, salts that can be used in intravenous pharmaceutical compositions, but preferably does not contain salts other than water-soluble zinc salts and sodium chloride.

[0027] Formulation etc. The first intravenous pharmaceutical composition of the present invention is not particularly limited as long as it can be used in the form of an aqueous intravenous pharmaceutical composition (i.e., an aqueous solution of an intravenous pharmaceutical composition) at the time of use.

[0028] For example, an example of a formulation form of the first intravenous pharmaceutical composition of the present invention is an aqueous intravenous pharmaceutical composition that does not require dilution or dissolution when used (i.e., an aqueous solution of the intravenous pharmaceutical composition; straight type).

[0029] Another example of the formulation form of the first intravenous pharmaceutical composition of the present invention is a concentrated aqueous intravenous pharmaceutical composition (i.e., a concentrated aqueous solution of the intravenous pharmaceutical composition; concentrated type), which requires dilution with water or an aqueous solution (e.g., physiological saline) before use.

[0030] Yet another example of the formulation of the first intravenous pharmaceutical composition of the present invention is a solid intravenous pharmaceutical composition (i.e., a dried intravenous pharmaceutical composition; dry type) that requires dissolution with water or an aqueous solution (e.g., physiological saline) before use. Specific examples of the dried form include freeze-dried products and spray-dried products.

[0031] The product categories of the first pharmaceutical composition for intravenous injection of the present invention include, for example, injections and infusions, and preferably injections.

[0032] Uses The first intravenous pharmaceutical composition of the present invention is useful for treating hypoalbuminemia, a condition requiring zinc administration. The first intravenous pharmaceutical composition of the present invention is used for the purpose of supplementing zinc for hypoalbuminemia, a condition requiring zinc administration. Specific examples of hypoalbuminemia include hypoalbuminemia and hypozincemia, more specifically, hypoalbuminemia associated with liver diseases such as hepatitis B, hepatitis C, alcoholic fatty liver, non-alcoholic fatty liver, and cirrhosis; kidney diseases such as diabetes and nephrotic syndrome; collagen diseases, malignant tumors, malnutrition, inflammatory bowel disease, acute pancreatitis, pregnancy-induced hypertension, severe burns, and sepsis; and drug-induced hypoalbuminemia.

[0033] The hypoalbuminemia symptom requiring zinc administration may be accompanied by hyperammonemia, and the first pharmaceutical composition for intravenous injection of the present invention can be used for treating the hyperammonemia associated with the symptom.

[0034] The animal species to which the first pharmaceutical composition for intravenous injection of the present invention is applied is not particularly limited as long as it has albumin as a serum protein, but preferred examples include mammals such as humans, cows, horses, cats, and dogs.

[0035] The method of administration of the first pharmaceutical composition for intravenous administration of the present invention is not limited as long as it is administered intravenously, and may be by injection or drip infusion. Among these methods, injection is preferred.

[0036] The dose of the first pharmaceutical composition for intravenous injection of the present invention can be determined depending on the degree of hypozincemia, the animal species, the age, etc., and may be, for example, 0.05 to 10 mg / kg / dose, preferably 0.1 to 5 mg / kg / dose, more preferably 0.3 to 3 mg / kg / dose, even more preferably 0.5 to 1.5 mg / kg / dose, and even more preferably 0.6 to 1.2 mg / kg / dose, 0.6 to 0.8 mg / kg / dose, or 0.8 to 1.2 mg / kg / dose in terms of the amount of zinc ions in humans.

[0037] The first pharmaceutical composition for intravenous injection of the present invention is preferably administered multiple times. The number of administrations in the repeated administration is not particularly limited, and examples include 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more. The upper limit of the number of administrations in the repeated administration is not particularly limited, and examples include 20 or less, 15 or less, 12 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or more, or 5 or less. Specific ranges of the number of administrations in the repeated administration include, for example, 2 to 20 times, 3 to 15 times, 4 to 12 times, 5 to 12 times, 6 to 12 times, 7 to 12 times, 8 to 12 times, 9 to 12 times, 10 to 12 times, 4 to 10 times, 4 to 9 times, 4 to 8 times, 4 to 7 times, 4 to 6 times, or 4 to 5 times. The administration frequency in repeated administration is not particularly limited, but examples include once every 2 to 14 days, preferably once every 3 to 12 days, more preferably once every 4 to 10 days, even more preferably once every 5 to 9 days, even more preferably once every 6 to 8 days, and particularly preferably once every 7 days.

[0038] The first pharmaceutical composition for intravenous injection of the present invention can be produced by any method that allows zinc ions and serum albumin to form a conjugate in the composition. Specifically, the first pharmaceutical composition for intravenous injection can be obtained by dissolving a water-soluble zinc salt, serum albumin, and other ingredients, if necessary, in water or physiological saline, stirring the mixture at room temperature for 10 minutes or more, preferably 60 minutes or more, and then concentrating and / or drying the mixture according to the desired formulation, if necessary.

[0039] 2. Second Intravenous Pharmaceutical Composition The second intravenous pharmaceutical composition of the present invention is characterized by containing zinc ions. The second intravenous pharmaceutical composition of the present invention will be described in detail below.

[0040] Zinc Ion Source A water-soluble zinc salt can be used as the zinc ion source. The water-soluble zinc salt is not particularly limited as long as it is a pharmaceutically acceptable salt. Examples of water-soluble zinc salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, and sulfuric acid; and salts of organic acids such as acetic acid, citric acid, tartaric acid, lactic acid, succinic acid, fumaric acid, maleic acid, and methanesulfonic acid. The water-soluble zinc salt may be an anhydride or a hydrate.

[0041] In the second pharmaceutical composition for intravenous injection of the present invention, the water-soluble zinc salt may be one of the above salts alone or a combination of two or more thereof. Among the above salts, from the viewpoint of further improving the efficiency of zinc supplementation, salts of inorganic acid salts are preferred, sulfate salts (i.e., zinc sulfate) are more preferred, and zinc sulfate heptahydrate is even more preferred.

[0042] The zinc ion content may be appropriately determined depending on the severity of iron overload and / or the desired level of iron-reducing effect, and / or the need for dilution or dissolution before use. For example, the zinc ion concentration in the pharmaceutical composition for intravenous administration at the time of use may be, for example, 0.1 mM or more, preferably 0.5 mM or more, more preferably 0.7 mM or more, even more preferably 0.8 mM or more, and even more preferably 0.9 mM or more. The upper limit of the zinc concentration is not particularly limited, and may be, for example, 5 mM or less, preferably 4 mM or less, more preferably 3 mM or less, even more preferably 2 mM or less, even more preferably 1.5 mM or less, and even more preferably 1.2 mM or less. Specific ranges of the zinc ion concentration in the pharmaceutical composition for intravenous administration at the time of use include, for example, 0.1 to 5 mM or 0.1 to 4 mM, preferably 0.5 to 3 mM, more preferably 0.7 to 2 mM, even more preferably 0.8 to 1.5 mM, and even more preferably 0.9 to 1.2 mM.

[0043] Serum Albumin The second intravenous pharmaceutical composition of the present invention may or may not contain serum albumin. For example, when the subject to which the second intravenous pharmaceutical composition of the present invention is applied has iron overload accompanied by hypoalbuminemia, it is preferable that the second intravenous pharmaceutical composition of the present invention contains serum albumin, since this has a superior iron-reducing effect.

[0044] When the second pharmaceutical composition for intravenous injection of the present invention contains serum albumin, the origin of the serum albumin is not particularly limited as long as it is the same as that of the animal species to which the pharmaceutical composition for intravenous injection is to be administered.

[0045] The content of serum albumin may be appropriately determined depending on the degree of iron overload and / or the desired level of iron-reducing effect, and may be, for example, 0.35 mol or more, preferably 0.4 mol or more, more preferably 0.5 mol or more, or 0.55 mol or more, relative to 1 mol of zinc ion. The upper limit of the content of serum albumin relative to 1 mol of zinc ion is not particularly limited, and may be, for example, 1 mol or less, preferably 0.9 mol or less, more preferably 0.8 mol or less, and even more preferably 0.7 mol or less, or 0.65 mol or less. Specific ranges of the content of serum albumin relative to 1 mol of zinc ion include, for example, 0.35 to 0.9 mol, preferably 0.4 to 0.8 mol, more preferably 0.5 to 0.7 mol, or 0.55 to 0.65 mol.

[0046] Forms of Zinc Ions and Serum Albumin When the second pharmaceutical composition for intravenous injection of the present invention contains serum albumin, the zinc ions and serum albumin form a conjugate. Specifically, some or all of the zinc ions can form a conjugate with serum albumin. In the present invention, the term "zinc ions" refers to both free zinc ions and zinc ions conjugated with serum albumin. Furthermore, the amount of zinc ions refers to the total amount of free zinc ions and zinc ions conjugated with serum albumin. When the serum albumin is human serum albumin, the conjugate is considered to be an embodiment in which the zinc ions are bound between His67 and Asn99 in domain I and His247 and Asp249 in domain II of human serum albumin.

[0047]

[0042] In addition to the above-mentioned components, the second pharmaceutical composition for intravenous injection of the present invention may contain other base materials and / or additives that can be used in pharmaceutical compositions for intravenous injection. Such base materials typically include water, and additives include isotonic agents (typically sodium chloride) and pH adjusters.

[0048] The second intravenous pharmaceutical composition of the present invention preferably does not contain any nutrients that can be used in intravenous pharmaceutical compositions in addition to the above-mentioned components, but such nutrients include carbohydrates (glucose, fructose, xylitol, etc.), amino acids, vitamins, etc.

[0049] The second intravenous pharmaceutical composition of the present invention may contain, in addition to the above-mentioned components, salts that can be used in intravenous pharmaceutical compositions, but preferably does not contain salts other than water-soluble zinc salts and sodium chloride.

[0050] Formulation etc. The second intravenous pharmaceutical composition of the present invention is not particularly limited as long as it can be used in the form of an aqueous intravenous pharmaceutical composition (i.e., an aqueous solution of an intravenous pharmaceutical composition) at the time of use.

[0051] For example, an example of a formulation form of the second intravenous pharmaceutical composition of the present invention is an aqueous intravenous pharmaceutical composition that does not require dilution or dissolution when used (i.e., an aqueous solution of the intravenous pharmaceutical composition; straight type).

[0052] Another example of the formulation form of the second intravenous pharmaceutical composition of the present invention is a concentrated aqueous intravenous pharmaceutical composition (i.e., a concentrated aqueous solution of the intravenous pharmaceutical composition; concentrated type), which requires dilution with water or an aqueous solution (e.g., physiological saline) before use.

[0053] Yet another example of the formulation of the second intravenous pharmaceutical composition of the present invention is a solid intravenous pharmaceutical composition (i.e., a dried intravenous pharmaceutical composition; dry type) that requires dissolution with water or an aqueous solution (e.g., physiological saline) before use. Specific examples of the dried form include freeze-dried products and spray-dried products.

[0054] The product categories of the second pharmaceutical composition for intravenous injection of the present invention include, for example, injections and infusions, preferably injections.

[0055] The second intravenous pharmaceutical composition of the present invention is used to treat iron overload, i.e., to reduce the amount of iron in the body of a patient suffering from iron overload, thereby treating the condition.

[0056] Specific examples of iron overload include secondary iron overload due to blood transfusion, iron overload due to excessive intake of iron-containing tablets and injections, hereditary iron overload, and alcoholism. Iron overload may be accompanied by complications. These complications include cirrhosis, hypoalbuminemia, hyperammonemia, diabetes, and heart failure. Specific examples of hypoalbuminemia include liver diseases such as hepatitis B, hepatitis C, alcoholic fatty liver, nonalcoholic fatty liver, and cirrhosis; kidney diseases such as diabetes and nephrotic syndrome; collagen disease, malignant tumor, malnutrition, inflammatory bowel disease, acute pancreatitis, pregnancy-induced hypertension, severe burns, and sepsis; and drug-induced hypoalbuminemia.

[0057] Furthermore, since the second pharmaceutical composition for intravenous injection of the present invention contains water-soluble zinc as an active ingredient, the iron overload may be accompanied by symptoms requiring zinc administration, as described above in the section "Use" in "1. First pharmaceutical composition for intravenous injection."

[0058] The animal species to which the second intravenous pharmaceutical composition of the present invention is applied is not particularly limited as long as it has albumin as a serum protein, but preferred examples include mammals such as humans, cows, horses, cats, and dogs.

[0059] The method of administration of the second pharmaceutical composition for intravenous administration of the present invention is not limited as long as it is administered intravenously, and may be by injection or by drip infusion. Among these methods, injection is preferred.

[0060] The dose of the second pharmaceutical composition for intravenous injection of the present invention can be determined depending on the degree of iron overload, the animal species, the age, etc., and may be, for example, 0.05 to 10 mg / kg / dose, preferably 0.1 to 5 mg / kg / dose, more preferably 0.3 to 3 mg / kg / dose, even more preferably 0.5 to 1.5 mg / kg / dose, and even more preferably 0.6 to 1.2 mg / kg / dose, 0.6 to 0.8 mg / kg / dose, or 0.8 to 1.2 mg / kg / dose in terms of the amount of zinc ions for humans.

[0061] The second pharmaceutical composition for intravenous injection of the present invention is preferably administered multiple times. The number of administrations in the repeated administration is not particularly limited, and examples include 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more. The upper limit of the number of administrations in the repeated administration is not particularly limited, and examples include 20 or less, 15 or less, 12 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or more, or 5 or less. Specific ranges of the number of administrations in the repeated administration include, for example, 2 to 20 times, 3 to 15 times, 4 to 12 times, 5 to 12 times, 6 to 12 times, 7 to 12 times, 8 to 12 times, 9 to 12 times, 10 to 12 times, 4 to 10 times, 4 to 9 times, 4 to 8 times, 4 to 7 times, 4 to 6 times, or 4 to 5 times. The administration frequency in repeated administration is not particularly limited, but examples include once every 2 to 14 days, preferably once every 3 to 12 days, more preferably once every 4 to 10 days, even more preferably once every 5 to 9 days, even more preferably once every 6 to 8 days, and particularly preferably once every 7 days.

[0062] Manufacturing Method The second pharmaceutical composition for intravenous injection of the present invention can be obtained, for example, by dissolving a water-soluble zinc salt, serum albumin which is blended as needed, and / or other ingredients which are blended as needed in water or physiological saline, stirring the mixture at room temperature for 10 minutes or more, preferably 60 minutes or more, and then concentrating and / or drying the mixture according to the formulation, as needed.

[0063] The present invention will be explained in more detail below by showing examples, but the present invention is not limited to these examples.

[0064] Test Example 1: First Intravenous Pharmaceutical Composition Test Example 1-1 [1] Materials Used Human serum-derived albumin (Fujifilm Wako Pure Chemical Industries, Ltd.) Zinc sulfate heptahydrate (Fujifilm Wako Pure Chemical Industries, Ltd.) Physiological saline (Otsuka Pharmaceutical Factory, Inc.) Dialysis cassette (material: regenerated cellulose, molecular weight cutoff: 10K, Thermo Scientific Corporation, product number (catalog number): 66455)

[0065] [2] Preparation of Intravenous Solutions Zinc sulfate heptahydrate was dissolved in physiological saline to a concentration of 1.0 mM, and human serum albumin was further added to the solution to give concentrations of 0 μM, 100 μM, 200 μM, 300 μM, 400 μM, 500 μM, or 600 μM (n=3 for each), followed by mixing at room temperature for 10 minutes to prepare intravenous solutions having the compositions of Comparative Example 1-1 and Examples 1-1a to 1-1f in Table 1. The pH of Example 1-1f at 25° C. was approximately 6.9.

[0066]

[0067] [3] Measurement of albumin-bound zinc ion concentration. The cassette was immersed in ultrapure water for 1 to 2 minutes for hydrophilization. 3 mL of each intravenous solution was collected in a syringe and injected through the port of the hydrophilized cassette using an 18G needle. After attaching a float to the cassette, the cassette was suspended in approximately 1000 mL of ultrapure water and dialyzed at room temperature for 2 hours. The cassette was then suspended again in approximately 1000 mL of fresh ultrapure water and dialyzed at room temperature for 2 hours. The cassette was then suspended in approximately 1000 mL of fresh ultrapure water and overnight dialysis was performed once at 4°C. After dialysis, the post-dialysis intravenous solution was collected from an uninserted port on the cassette using an 18G needle and syringe. Zinc in the post-dialysis intravenous solution was measured (direct method: 5-Br-PAPS) using an automated analyzer 7180 (Hitachi High-Tech Corporation). The results are shown in Figure 1.

[0068] As shown in Figure 1, the amount of zinc bound to albumin increased depending on the albumin concentration, and when the amount of albumin added reached 400 μM or more, the amount of zinc bound to albumin plateaued. In other words, it was found that when the albumin concentration was 400 μM or more per 1 mM zinc (0.4 mol or more per 1 mol of zinc), the albumin that can bind to zinc was saturated.

[0069] Test Example 1-2 The water-soluble zinc salt and albumin to be added to physiological saline were formulated as shown in Table 2 to prepare the intravenous solutions of Examples 1-2a to 1-2f.

[0070]

[0071] The prepared intravenous solution was subjected to the method described in [3] of Test Example 1-1, and the zinc content in the intravenous solution after dialysis (direct method: 5-Br-PAPS) and albumin content (BCG method) were measured. The measured zinc concentration (Zn) was divided by the measured albumin concentration (ALB) to determine the value (Zn / ALB). A larger Zn / ALB value indicates a stronger binding strength of zinc ions to albumin. The results are shown in Figure 2.

[0072] As a result, binding of zinc ions to albumin was observed in all Examples, and as shown in Figure 2, it was found that the binding strength of zinc ions to albumin was greater in Examples 1-2e and 1-2f than in Examples 1-2a to 1-2d. It has been suggested that albumin is a carrier through which hepatocytes acquire zinc in vivo, so it can be said that the stronger the binding strength of a zinc salt to albumin, the greater the zinc supplementation effect to the liver. Therefore, it can be reasonably inferred that zinc sulfate has a greater zinc supplementation effect to the liver than zinc acetate and chloride salts.

[0073] Test Example 1-3 [1] Materials used Human serum-derived albumin (Fujifilm Wako Pure Chemical Industries, Ltd.) Zinc sulfate heptahydrate (Fujifilm Wako Pure Chemical Industries, Ltd.) Physiological saline (Otsuka Pharmaceutical Factory, Inc.)

[0074] [2] Preparation of Test Intravenous Solutions <Examples> Human serum-derived albumin and zinc sulfate heptahydrate were blended into physiological saline solution to give concentrations of 600 μM and 1.0 mM (0.6 mol of albumin per 1 mol of zinc), respectively, and stirred with a stirrer at room temperature for approximately 30 minutes to prepare the test intravenous solutions (also referred to as "Zn-Alb") of Examples 1-3. <Comparative Examples> Zinc sulfate heptahydrate was blended into physiological saline solution to give a concentration of 1.0 mM, and stirred with a stirrer at room temperature for approximately 30 minutes to prepare the test intravenous solution (also referred to as "Zn") of Comparative Example 3.

[0075]

[0076] [3] Animals used and rearing conditions: NAR / Slc female (8-9 weeks old) and male (6-7 weeks old) rats (Japan SLC Co., Ltd.) were used. This strain is a mutant rat that exhibits analbuminemia due to autosomal recessive inheritance and is used as a model for patients with hypoalbuminemia.

[0077] These rats were fed a zinc-deficient diet (Oriental Yeast Co., Ltd.) for 7 weeks to create a hypoalbuminemia and hypozincemia model. During the feeding period, the rats were allowed free access to ultrapure water (changed twice a week) from a water bottle.

[0078] [4] Administration of Test Intravenous Solution [4-1] Single Administration: The administration site was the tail vein, the number of administrations was single, and the dose was 5 mL / kg. This dose was 0.325 mg / kg zinc, equivalent to a 16.3 mg dose in a 50 kg human. The day of the single administration was counted as Day 0 (Day 0), and the day following the single administration was counted as Day 1. [4-2] Repeat Administration: The administration site was the tail vein, the number of administrations was 8 (once per week), and the dose was 10 mL / kg. This dose was 0.65 mg / kg zinc, equivalent to a 32.6 mg dose in a 50 kg human. The day of the first repeated administration was counted as Day 28 (Day 28), and the day following the first repeated administration was counted as Day 29 (Day 29). [4-3] Administration Method: The animals were immobilized using a restrainer. Under no anesthesia, a sterilized indwelling needle (24 gauge, Surflow F&F, Terumo Corporation) was inserted into the tail vein and left there. The test substance was collected in a syringe and administered via the catheter using a syringe. The administration rate was approximately 2.0 mL / kg / min.

[0079] [5] Serum Zinc Concentration Measurement and Results [5-1] Single-Dose Administration Measurements were performed in the afternoon of the day before the single administration and 1, 7, 14, 21, and 28 days after the single administration. Animals were manually immobilized, and 300 μL of blood was collected from the jugular vein without anesthesia. Blood was transferred to a tube containing a coagulation promoter / serum separator (Microtina micro-blood collection tube, Becton, Dickinson, Japan). After leaving the tube at room temperature for 30-60 minutes, serum was obtained by centrifugation (3000 rpm, 10 minutes, 4°C) in a floor-type refrigerated centrifuge S700FR. Serum zinc concentrations were measured using an automated analyzer 7180 (Hitachi High-Tech Corporation). The results are shown in Figure 3A. Furthermore, the zinc increase was calculated by subtracting the zinc concentration (zinc value) on the day before the single administration from the zinc concentration (zinc value) on days 1, 7, 14, 21, and 28 after the single administration. The results are shown in Figure 3B.

[0080] [5-2] Repeated Administration Measurements were performed in the afternoon 14 days (2 weeks), 28 days (4 weeks), 42 days (6 weeks), and 56 days (8 weeks) after the initial repeated administration, and before the repeated administration on the same day (i.e., 7 days after the previous administration). Animals were manually immobilized, and 300 μL of blood was collected from the jugular vein without anesthesia. The blood was transferred to a tube containing a coagulation promoter / serum separator (Microtina micro-collection tube, Becton, Dickinson, Japan). After leaving the tube at room temperature for 30-60 minutes, serum was obtained by centrifugation (3000 rpm, 10 minutes, 4°C) in a floor-type refrigerated centrifuge S700FR. Serum zinc concentrations were measured using an automated analyzer 7180 (Hitachi High-Tech Corporation). The results are shown in Figure 4A. Furthermore, the zinc increase was calculated by subtracting the zinc concentration (zinc value) on the day of the first repeated administration from the zinc concentration (zinc value) 14 days (2 weeks), 28 days (4 weeks), 42 days (6 weeks), and 56 days (8 weeks) after the first repeated administration. The results are shown in Figure 4B.

[0081] [5-3] Results As shown in the results for "Day 7" in Figures 3A and 3B, a transient decrease in serum zinc concentration was observed in patients receiving the test intravenous solution (Zn-Alb) of Example 1-3 compared to patients receiving the test intravenous solution (Zn) of Comparative Example 1-3. The dynamics of zinc in the body are such that absorbed zinc is first transported to the liver and then distributed throughout the body (Guidelines for the Treatment of Zinc Deficiency 2018, Japan Society of Clinical Nutrition, 2018, p. 1-46). Therefore, the low serum zinc concentration observed on "Day 7" in Figures 3A and 3B reasonably indicates that this fluctuation reflects the high bioabsorbability of the novel zinc formulation.

[0082] Furthermore, in repeated administration (measurement results 7 days after the previous administration), as shown in Figure 4B, a stable increase in zinc was observed when the test intravenous solution (Zn-Alb) of Example 1-3 was administered compared to when the test intravenous solution (Zn) of Comparative Example 1-3 was administered, and as shown in Figure 4A, after 8 weeks, the blood zinc concentration converted to a higher value when the test intravenous solution (Zn-Alb) of Example 1-3 was administered compared to when the test intravenous solution (Zn) of Comparative Example 1-3 was administered. The result of conversion of serum zinc to a higher value in repeated administration (measurement results 7 days after the previous administration) reasonably indicates that zinc taken up into the liver by the previous administration was redistributed into the blood.

[0083] Test Example 1-4 [1] Preparation of Materials Used and Test Intravenous Solutions Using the materials used in Test Example 1-3, the zinc source and albumin to be mixed into physiological saline were formulated as shown in Table 4 to prepare intravenous solutions of Comparative Example 1-4 and Examples 1-4a to 1-4c.

[0084]

[0085] [2] Animals used and rearing conditions The same animals and rearing conditions were used as in Test Example 1-3.

[0086] [3] Administration of Test Intravenous Solution [3-1] Repeated Administration The administration site was the tail vein, the number of administrations was 10 (once per week), and the dose was 15 mL / kg. This dose amounted to 0.975 mg / kg / administration of zinc, equivalent to 48.9 mg / administration for a 50 kg human, and approximately 14 times the ASPEN recommended daily zinc intake of 3-4 mg in parenteral nutrition. The first through tenth administrations are referred to as Dose 1 to Dose 10, respectively. [3-2] Administration Method The animal was immobilized using a restrainer. A sterilized indwelling needle (24 gauge, Surflow F&F, Terumo Corporation) was inserted and left in the tail vein without anesthesia. The test substance was collected in a syringe and administered via a catheter using a syringe. The administration rate was approximately 4.0 mL / kg / min.

[0087] [4] Blood Biochemistry Tests and Results Tests were conducted in the morning following each repeated administration (i.e., 1 day after the previous administration). Animals were manually immobilized, and 500 μL of blood was collected from the jugular vein without anesthesia. The blood was transferred to a tube containing a coagulation promoter / serum separator (Microtina micro-blood collection tube, Becton, Dickinson, Japan Co., Ltd.) and left at room temperature for 30-60 minutes. The serum obtained by centrifugation (3000 rpm, 10 minutes, 4°C) in a refrigerated centrifuge was used for testing. The serum was measured using an automated analyzer BM6050. The amount of ammonia reduction (Serum NH3 red) was calculated as the amount of ammonia reduction compared to the day before the first administration, using the ammonia measured at each blood collection time and the day before the first administration.

[0088] The mean zinc and albumin levels in each group on the day before the first administration were 65 μg / dL and 1.3 g / dL or less, respectively, confirming that this model was established as a hypoalbuminemia-hypozincemia model animal. Furthermore, the mean ammonia levels in each group exceeded 135 μg / dL, confirming that this model reflected the pathological condition of hyperammonemia. The ammonia measurement results following repeated administration of Comparative Example 1-4 (Zn) and Examples 1-4a to 1-4c (Zn-Alb 200, 400, 600) are shown in Figure 5A.

[0089] As shown in Figure 5A, administration of the intravenous solutions of Examples 1-4a to 1-4c (Zn-Alb200, 400, 600) significantly reduced ammonia levels (the degree of ammonia reduction was significantly increased) compared with administration of the intravenous solution of Comparative Example 1-4 (Zn), suggesting therapeutic efficacy for hyperammonemia associated with hypoalbuminemia and hypozincemia. Furthermore, in the urea cycle, which is central to ammonia metabolism in the body, ornithine transcarbamylase (OTC), a zinc enzyme, functions within hepatocytes. The hyperammonemia observed in the background values ​​of the model animals was the result of inhibition of OTC enzyme activity due to zinc deficiency. The albumin concentration-dependent ammonia metabolic effect observed after administration is thought to be the result of zinc being efficiently transported to the liver by albumin, a result that complements the high zinc supplementation effect of the intravenous solutions of Examples 1-4a to 1-4c (Zn-Alb200, 400, 600).

[0090] [5] Measurement of liver zinc concentration and results [5-1] Measurement method After blood collection, the animals were sacrificed by exsanguination, and the livers were collected. The tissues were frozen and stored at -80°C and used as samples. 10 mL of 0.1 N hydrochloric acid was added to 0.5 g of each thawed sample. After homogenization, the sample was allowed to stand for 30 minutes, and the supernatant was centrifuged (10,000 rpm, 10 min, 4°C). The resulting supernatant was used as an assay sample, and the absorbance at 560 nm was measured using a Metalloassay Zinc Measurement LS (Metalogenix Inc.) and a microplate reader, and the zinc concentration was calculated.

[0091] [5-2] Results The results are shown in Figure 5B. As shown in Figure 5B, compared to the administration of the intravenous solution of Comparative Example 1-4 (Zn), the administration of the intravenous solutions of Examples 1-4a to 1-4c (Zn-Alb200, 400, 600) resulted in increased zinc levels in the liver. Since zinc binds to albumin in the blood and is transported to organs throughout the body, the increased zinc levels reflect the tissue transfer effect of zinc preparations containing albumin as a zinc carrier, i.e., the high zinc supplementation effect to the liver. As shown in Figure 5B, Example 1-4b (Zn-Alb400) showed the highest amount of zinc accumulation in the liver. However, in light of the zinc supplementation results shown by the ammonia metabolism effect in Figure 5A of this test example and the zinc redistribution effect in the blood in Figures 4A and 4B of Test Example 1-3, Example 1-4c (Zn-Alb600) can be evaluated as the preparation with the best zinc supplementation effect, which stabilizes zinc in the blood through accumulation and redistribution in the liver.

[0092] Test Example 2: Second Intravenous Pharmaceutical Composition Test Example 2-1 (Preliminary Test on In vivo Iron Removal Effect) [1] Materials Used and Preparation of Test Samples The following components were mixed with physiological saline to the contents shown in the table, and the mixture was stirred with a stirrer at room temperature for about 10 minutes to prepare the test samples of Examples 2-1a, 2-1b, and Reference Example.

[0093]

[0094] [2] Animals used and rearing conditions Eighteen male (6-7 weeks old) Crl:(CD)SD rats (Jackson Laboratory Japan) were used.

[0095] During the test period, the rats were given free access to standard feed CRF-1 (Oriental Yeast Co., Ltd.) and tap water passed through a sterilized filter.

[0096] [3] Test Sample Administration In Examples 2-1a and 2-1b, administration was via the tail vein (administration rate: 4.0 mL / kg / min), while in the Reference Example, oral administration (administration via a probe) was performed. The number of administrations was seven, with the administration frequency being once per day. The dosages are as shown in Table 5. The dosages in Examples 2-1a and 2-1b were 0.975 mg / kg of zinc, which corresponds to 48.75 mg in a human weighing 50 kg. The dosage of deferasirox (Novartis Pharma K.K.'s "Jadonnu," a drug for treating iron overload) in the Reference Example was the upper limit of the clinical dosage.

[0097] [4] Blood Biochemistry Tests and Results Blood biochemistry tests were performed on all animals on the day of grouping and the day after the final administration. Blood biochemistry tests were performed from the tail vein and the caudal vena cava, respectively, and at least 300 μL was collected for blood biochemistry tests. All blood samples were collected after fasting for at least 16 hours from the previous day. The collected blood was quickly transferred to a microtube containing heparin and mixed. Plasma was collected by centrifugation (1900 × g, 10 minutes, 4°C), and plasma iron (bathophenanthroline direct method), unsaturated iron binding capacity (UIBC) (nitroso-PSAP method), and ferritin (ELISA method) were measured. The iron and ferritin levels were calculated by subtracting the values ​​measured on the day after the final administration from the values ​​measured on the day of grouping to determine the decrease in iron and ferritin levels. The results are shown in Figures 6A and 6B. The unsaturated iron binding capacity was calculated by subtracting the values ​​measured on the day after the final administration from the values ​​measured on the day of grouping to determine the increase in unsaturated iron binding capacity. The results are shown in Figure 6C.

[0098] The ferritin-reducing effect, iron-reducing effect, and UIBC-increasing effect shown in Figures 6A to 6C were shown to be at the same level as those of deferasirox (Reference Example), a known therapeutic drug for iron overload, for both the intravenous solution of Zn (Example 2-1a) and the intravenous solution of Zn-Alb (Example 2-1b).

[0099] Test Example 2-2 (In vivo iron removal effect in iron-loaded rats) [1] Materials used and preparation of intravenous solutions The following components were mixed with physiological saline to the contents shown in the table to prepare intravenous solutions of Comparative Example 2-2, Example 2-2a, and Example 2-2b.

[0100]

[0101] [2] Animals Used and Rearing Conditions Twenty-four male (6-7 weeks old) Crl:(CD)SD rats (Jackson Laboratory Japan, Inc.) were used. Under unanesthetized conditions, iron loading was performed by subcutaneously administering a single dose of iron dextran solution (Sigma-Aldrich Japan GK) (400 mg / kg) to two sites on the back using a needle and syringe to create an iron overload (hemochromatosis) model.

[0102] During the test period, the rats were given free access to standard feed CRF-1 (Oriental Yeast Co., Ltd.) and tap water passed through a sterilized filter.

[0103] [3] Intravenous administration Starting from the third week after iron loading, Comparative Example 2-2, Example 2-2a, and Example 2-2b were administered via the tail vein (administration rate: 4.0 mL / kg / min). The number of administrations was 14, and the administration frequency was once per day. As shown in Table 6, the dose was approximately three times the dose in Test Example 2-1. This dose was 3.25 mg / kg of zinc, which corresponds to a dose of 162.5 mg for a human weighing 50 kg.

[0104] [4] Blood Biochemistry Tests and Results Blood biochemistry tests were performed on all subjects three weeks after iron loading and the day after the final administration. Blood samples were taken from the jugular vein and the posterior vena cava, respectively, and 500 μL or more was collected for blood biochemistry tests. All blood samples were taken after fasting for at least 16 hours from the day before. Plasma iron was measured in the same manner as in Test Example 2-1, and the amount of iron reduction was calculated by subtracting the measurement value measured the day after the final administration from the measurement value measured three weeks after iron loading. The results are shown in Figure 7.

[0105] 7, both the intravenous solution of Zn-Sul (Example 2-2a) and the intravenous solution of Zn-Alb (Example 2-2b) were found to have an iron-reducing effect on the iron overload model. Taking into account the comparison results of deferasirox (Reference Example) with Zn (Example 2-1a) and Zn-Alb (Example 2-1b) in Test Example 2-1, it can be inferred that both the intravenous solution of Zn-Sul (Example 2-2a) and the intravenous solution of Zn-Alb (Example 2-2b) have a therapeutic effect on iron overload equivalent to that of deferasirox.

[0106] Test Example 2-3 [1] Preparation of materials used and test intravenous solutions Using the materials used in Test Example 1-3, the zinc source and albumin to be mixed in physiological saline were formulated as shown in Table 7, and the intravenous solutions of Examples 2-3a to 2-3c were prepared.

[0107]

[0108] [2] Animals used and rearing conditions The same animals and rearing conditions were used as in Test Example 1-3.

[0109] [3] Administration of Test Intravenous Solution [3-1] Repeated Administration The administration site was the tail vein, the number of administrations was 10 (once per week), and the dose was 15 mL / kg. This dose amounted to 0.975 mg / kg / administration of zinc, equivalent to 48.9 mg / administration for a 50 kg human, and approximately 14 times the ASPEN recommended daily zinc intake of 3-4 mg in parenteral nutrition. The first through tenth administrations are referred to as Dose 1 to Dose 10, respectively. [3-2] Administration Method The animal was immobilized using a restrainer. A sterilized indwelling needle (24 gauge, Surflow F&F, Terumo Corporation) was inserted and left in the tail vein without anesthesia. The test substance was collected in a syringe and administered via a catheter using a syringe. The administration rate was approximately 4.0 mL / kg / min.

[0110] [4] Blood Biochemistry Tests and Results Tests were conducted in the morning following each repeated administration (i.e., 1 day after the previous administration). Animals were manually immobilized, and 500 μL of blood was collected from the jugular vein without anesthesia. The blood was transferred to a tube containing a coagulation promoter / serum separator (Microtina micro-blood collection tube, Becton, Dickinson, Japan Co., Ltd.) and left at room temperature for 30-60 minutes. The serum obtained by centrifugation (3000 rpm, 10 minutes, 4°C) in a refrigerated centrifuge was used for testing. Serum was measured using an automated analyzer BM6050. Iron loss (Serum Fe red) was calculated as the loss from the day before the first administration using iron measurements at each blood collection time and the day before the first administration.

[0111] The average zinc and albumin levels in each group on the day before the first administration were 65 μg / dL and 1.3 g / dL or less, respectively, confirming that this model was established as a hypoalbumin-hypozincemia animal model. Furthermore, the average ammonia levels in each group exceeded 135 μg / dL, confirming that this model reflected the pathology of hyperammonemia. Furthermore, the high iron measurement values ​​confirmed that this model also reflected the pathology of iron overload. The iron measurement results following repeated administration of Example 2-3a (Zn), Example 2-3b (Zn-Alb200), and Example 2-3c (Zn-Alb600) are shown in Figure 8.

[0112] As shown in Figure 8, repeated administration of Example 2-3a (Zn), Example 2-3b (Zn-Alb200), and Example 2-3c (Zn-Alb600) confirmed a reduction in iron levels. Furthermore, compared with administration of the intravenous solution of Example 2-3a (Zn), the intravenous solutions of Example 2-3b (Zn-Alb200) and Example 2-3c (Zn-Alb600) tended to result in a greater reduction in iron levels, and this was particularly evident in the intravenous solution of Example 2-3c (Zn-Alb600). In other words, the iron clearance effect on iron overload associated with hypoalbuminemia-hypozincemia was shown to be higher in intravenous solutions containing water-soluble zinc and albumin in accordance with the albumin content ratio.

Claims

1. A pharmaceutical composition for intravenous administration, comprising zinc ions and serum albumin.

2. The pharmaceutical composition for intravenous injection according to claim 1, wherein the zinc ion and the serum albumin form a conjugate.

3. The intravenous pharmaceutical composition of claim 1, wherein said zinc ion source is a water-soluble zinc salt.

4. The intravenous pharmaceutical composition of claim 3, wherein said water-soluble zinc salt is zinc sulfate.

5. The pharmaceutical composition for intravenous injection according to claim 1, wherein the content of said serum albumin per 1 mole of said zinc ion is 0.35 moles or more.

6. The pharmaceutical composition for intravenous injection described in claim 1, wherein the zinc ion content at the time of use is 0.1 mM or more.

7. The intravenous pharmaceutical composition according to claim 1, which does not contain carbohydrates.

8. The pharmaceutical composition for intravenous administration according to claim 1, which is an injection.

9. The pharmaceutical composition for intravenous administration according to claim 1, which is used for the treatment of symptoms of hypoalbuminemia requiring administration of zinc.

10. The pharmaceutical composition for intravenous administration according to claim 9, wherein the symptom is hypoalbuminemia and hypozincemia.

11. The pharmaceutical composition for intravenous administration according to claim 9, which is used for hyperammonemia.

12. An intravenous pharmaceutical composition containing zinc ions and used to treat iron overload.

13. The intravenous pharmaceutical composition of claim 12, wherein said zinc ion source is a water-soluble zinc salt.

14. The intravenous pharmaceutical composition of claim 13, wherein said water-soluble zinc salt is zinc sulfate.

15. The pharmaceutical composition for intravenous administration described in claim 12, wherein the zinc ion content at the time of use is 0.1 mM or more.

16. The pharmaceutical composition for intravenous administration according to claim 12, which is an injection.

17. The pharmaceutical composition for intravenous administration according to claim 12, further comprising serum albumin.

18. The intravenous pharmaceutical composition of claim 17, wherein said zinc ions and said serum albumin are conjugated.

19. The pharmaceutical composition for intravenous administration according to claim 17, wherein the content of said serum albumin per 1 mole of said zinc ion is 0.35 moles or more.

20. The intravenous pharmaceutical composition of claim 17, wherein the iron overload is accompanied by hypoalbuminemia.

21. The pharmaceutical composition for intravenous administration according to claim 20, wherein said hypoalbuminemia is hypoalbuminemia.

22. The intravenous pharmaceutical composition of claim 12, wherein the iron overload is accompanied by hyperammonemia.

23. The intravenous pharmaceutical composition of claim 12, which is carbohydrate-free.

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