Use of a composition comprising a zinc salt and a cyclic histidine-proline as an effective ingredient for preventing, ameliorating or treating diabetic nephropathy

Abstract

The present invention relates to a use of a composition comprising a zinc salt; and cyclohistidine-proline as an effective ingredient for preventing, ameliorating or treating diabetic nephropathy, and more particularly, to provide a composition comprising a zinc salt; and cyclohistidine-proline as an effective ingredient and a method for preventing, ameliorating or treating diabetic nephropathy using the same, wherein the composition exhibits a preventive, ameliorative or therapeutic effect on diabetic nephropathy through an effect of reducing albuminuria, reducing an albumin / creatinine ratio or increasing glomerular filtration rate when the composition is administered alone or in combination with an antihypertensive agent and / or an antidiabetic agent.

Description

Technical Field

[0001] This application claims priority to Korean Patent Application No. 10-2023-0153151, filed on November 7, 2023, the entire contents of which are disclosed in the specification and drawings of which are incorporated herein by reference.

[0002] This invention relates to the use of a composition comprising a zinc salt and cyclic histidine-proline as active ingredients for the prevention, improvement, or treatment of diabetic nephropathy. More specifically, it provides a composition comprising a zinc salt and cyclic histidine-proline as active ingredients and a method of using the same to prevent, improve, or treat diabetic nephropathy, wherein the composition, when administered alone or in combination with antihypertensive agents and / or antidiabetic agents, exhibits preventive, improving, or therapeutic effects on diabetic nephropathy through reductions in albuminuria, reductions in the albuminuria / creatinine ratio, or increases in glomerular filtration rate. Background Technology

[0003] Diabetic kidney disease (DKD) is a complication of diabetes, and it is estimated that approximately 40% of diabetic patients will develop DKD (Non-Patent Literature 1). Although the etiology of DKD is not fully understood, it is generally believed that it is caused by damage to blood vessels, cells, and tissues in the kidneys due to persistent hyperglycemia and hypertension, obesity, high-salt diet, and genetic factors (Non-Patent Literature 2). Kidney damage caused by DKD leads to proteinuria (albuminuria) and reduces the glomerular filtration rate (gFR), thereby impairing the important functions of the kidneys, namely filtration and reabsorption, and becoming a major cause of end-stage renal failure. Currently, for the prevention and treatment of disseminated intravascular coagulation (DKD), angiotensin II receptor blocker (ARB) losartan is being used to regulate blood pressure, and sodium-glucose cotransporter 2 (SGLT2) inhibitors, such as dapagliflozin, are used to regulate blood sugar. However, losartan, classified as an angiotensin II receptor blocker, can cause common side effects such as indigestion, diarrhea, and abdominal pain; while dapagliflozin, the SGLT2 inhibitor with the most clinical results, has been reported to frequently cause side effects such as headache, diarrhea, back pain, bronchitis, pharyngitis, and upper respiratory tract infections. Furthermore, conventional treatments that only regulate blood pressure and blood sugar are unlikely to slow or stop the progression of DKD. Therefore, there is a need to develop a strategic drug to improve treatment efficacy through the synergistic effect of developing new drugs or combining them with existing drugs.

[0004] On the other hand, Patent Document 1 discloses a composition containing zinc ions and cyclo-hispro (CHP) as a composition that helps alleviate the symptoms of diabetes in mammals, but does not disclose any examples of using an antihypertensive agent and / or an antidiabetic agent in combination in the composition for the purpose of preventing, improving or treating DKD.

[0005] Against this backdrop, the inventors have confirmed that the combination of zinc and cyclic histidine-proline not only exhibits preventive, ameliorative, or therapeutic effects on diabetic nephropathy through the reduction of albuminuria, the reduction of albuminuria / creatinine ratio, or the increase of glomerular filtration rate, but also significantly enhances the therapeutic effects of existing antihypertensive and / or antidiabetic agents for diabetic nephropathy when used in combination with them, thus completing this invention.

[0006] [Existing Technical Documents] [Patent Literature] (Patent Document 1) Korean Patent Publication No. 10-2001-0022786 [Non-patent literature] (Non-patent literature 1) Clin J Am Soc Nephrol: CJASN., 12 (2017), pp. 2032-204 (Non-patent literature 2) Int J Mol Sci. 2020 Mar 23;21(6):2218 Summary of the Invention The technical problem that the invention aims to solve Therefore, the object of the present invention is to provide a composition comprising a zinc salt and a cyclic histidine-proline combination for the prevention, improvement or treatment of diabetic nephropathy.

[0007] Furthermore, the present invention aims to provide a composition comprising a zinc salt and a cyclic histidine-proline combination for improving the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy.

[0008] Furthermore, the present invention aims to provide a method for preventing, improving, or treating diabetic nephropathy using zinc salts and a cyclic histidine-proline combination.

[0009] Furthermore, the present invention aims to provide a method for improving the preventive, therapeutic, or therapeutic effects of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy by utilizing zinc salts and cyclic histidine-proline combinations.

[0010] In addition, the present invention aims to provide the use of a composition comprising a zinc salt and a cyclic histidine-proline combination for the preparation of a medicament for the prevention, improvement or treatment of diabetic nephropathy.

[0011] Furthermore, the present invention aims to provide the use of a composition comprising a zinc salt and a cyclic histidine-proline combination for preparing a medicament for improving the preventive, ameliorative, or therapeutic effects of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy.

[0012] means for solving problems To address the aforementioned technical problems, the present invention provides a pharmaceutical composition for the prevention or treatment of diabetic nephropathy, comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

[0013] The present invention also provides a pharmaceutical composition for improving the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

[0014] The present invention also provides a health functional food composition for preventing or improving diabetic nephropathy, comprising zinc salt; and cyclo-hispro or its food-acceptable salt as an active ingredient.

[0015] In this invention, the pharmaceutical composition may include an antihypertensive agent selected from the group consisting of angiotensin II receptor blockers (ARBs) and angiotensin-converting enzyme inhibitors (ACE inhibitors) as an additional active ingredient.

[0016] In this invention, the pharmaceutical composition may include an antidiabetic agent selected from the group consisting of sodium-glucose cotransporter 2 inhibitor (SGLT2 inhibitor), dipeptidylpeptidase-4 inhibitor (DPP-4 inhibitor), and biguanide drugs as an additional active ingredient.

[0017] In this invention, the pharmaceutical composition may include an antihypertensive agent selected from the group consisting of ARBs and ACE inhibitors, and an antidiabetic agent selected from the group consisting of SGLT2 inhibitors, DPP-4 inhibitors, and biguanides as additional active ingredients.

[0018] In this invention, the ARB may be any one selected from the group consisting of Losartan, Valsartan, Candesartan, Olmesartan, Telmisartan, Fimasartan, Irbesartan, Eprosartan, and Azilsartan, and the ACE inhibitor may be selected from Captopril. It is any one of the following groups: pril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, ciliapril, and fosinopril.

[0019] In this invention, the SGLT2 inhibitor may be selected from any one of the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin, and the DPP-4 inhibitor may be selected from sitagliptin. The biguanide drug may be any one of the following groups: Sitagliptin, Linagliptin, Vildagliptin, Gemigliptin, Saxagliptin, Alogliptin, Tenegliptin, Anagliptin, and Evogliptin, wherein the biguanide drug may be any one of the following groups: metformin, buformin, and phenformin.

[0020] In this invention, the composition may exhibit one or more effects selected from the group consisting of: i) to iii) a reduction in albuminuria; ii) a reduction in the albuminuria / creatinine ratio; and iii) an increase in glomerular filtration rate.

[0021] In this invention, the composition may be administered simultaneously, alone, or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0022] In this invention, the health functional food composition can be used to improve the diabetic nephropathy effect in patients with diabetic nephropathy who are receiving the antihypertensive agent and / or the antidiabetic agent.

[0023] In addition, the present invention provides a method for the prevention, improvement or treatment of diabetic nephropathy, comprising the steps of: administering an effective amount of a zinc salt to an individual who requires the method; and a composition comprising cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0024] The present invention also provides a method for improving the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, comprising the steps of: administering an effective amount of a zinc salt to an individual who requires the method; and a composition comprising cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0025] Furthermore, the present invention provides the use of a composition for preparing a medicament for the prevention, improvement or treatment of diabetic nephropathy, said composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0026] In addition, the present invention provides the use of a composition for preparing a pharmaceutical agent for improving the preventive, ameliorative, or therapeutic effects of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, said composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0027] In this invention, the composition may include an antihypertensive agent selected from the group consisting of angiotensin II receptor blockers (ARBs) and angiotensin-converting enzyme inhibitors (ACE inhibitors) as an additional active ingredient, or may be administered in combination with the antihypertensive agent.

[0028] In this invention, the composition may include an antidiabetic agent selected from the group consisting of sodium-glucose cotransporter 2 inhibitor (SGLT2 inhibitor), dipeptidylpeptidase-4 inhibitor (DPP-4 inhibitor), and biguanide drugs as an additional active ingredient, or may be administered in combination with the antidiabetic agent.

[0029] In this invention, the composition may include an antihypertensive agent selected from the group consisting of ARBs and ACE inhibitors, and an antidiabetic agent selected from the group consisting of SGLT2 inhibitors, DPP-4 inhibitors and biguanides as an additional active ingredient, or may be used in combination with the antihypertensive agent and the antidiabetic agent.

[0030] In this invention, the ARB may be any one selected from the group consisting of Losartan, Valsartan, Candesartan, Olmesartan, Telmisartan, Fimasartan, Irbesartan, Eprosartan, and Azilsartan, and the ACE inhibitor may be selected from Captopril. It is any one of the following groups: pril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, ciliapril, and fosinopril.

[0031] In this invention, the SGLT2 inhibitor may be selected from any one of the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin, and the DPP-4 inhibitor may be selected from sitagliptin. The biguanide drug may be any one of the following groups: Sitagliptin, Linagliptin, Vildagliptin, Gemigliptin, Saxagliptin, Alogliptin, Tenegliptin, Anagliptin, and Evogliptin, wherein the biguanide drug may be any one of the following groups: metformin, buformin, and phenformin.

[0032] In this invention, the composition may exhibit one or more effects selected from the group consisting of: i) to iii) a reduction in albuminuria; ii) a reduction in the albuminuria / creatinine ratio; and iii) an increase in glomerular filtration rate.

[0033] In this invention, the composition may be administered simultaneously, alone, or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0034] Invention Effects The composition comprising zinc salt and cyclic histidine-proline according to the present invention, when administered alone, exhibits preventive, ameliorative, or therapeutic effects on diabetic nephropathy by reducing albuminuria, decreasing the albuminuria / creatinine ratio, or increasing glomerular filtration rate. Therefore, it can be used as a therapeutic agent and health functional food for the prevention, improvement, or treatment of diabetic nephropathy. Furthermore, even when used in combination with existing antihypertensive and / or antidiabetic agents prescribed to patients with diabetic nephropathy, it can significantly enhance the therapeutic efficacy of these drugs for diabetic nephropathy. Therefore, it can be useful as an adjunct to patients receiving the aforementioned drugs and can also help reduce side effects caused by chronic administration of these drugs. Attached Figure Description

[0035] Figures 1a to 1d To confirm the effect of CycloZ or various drug combinations on reducing albuminuria in KKAy mice, a model of diabetic nephropathy, the following steps were taken: Figure 1a This indicates the reduction in albuminuria resulting from CycloZ alone or in combination with Losartan (Losartan) + CycloZ. Figure 1b This indicates the reduction in albuminuria resulting from CycloZ alone or in combination with dapagliflozin. Figure 1c This indicates the reduction in albuminuria resulting from CycloZ alone or from a combination of Losartan, Dapagliflozin, and CycloZ. Figure 1d This indicates that the above Figures 1a to 1c The results are shown in a chart for comparison.

[0036] Figures 2a to 2c To confirm the effect of CycloZ or various drug combinations on reducing the albuminuria-to-creatinine ratio (ACR) in KKAy mice, a model of diabetic nephropathy, the following results were obtained: Figure 2aThis indicates the reduction in ACR caused by CycloZ alone or in combination with Losartan (Losartan) + CycloZ. Figure 2b This indicates the reduction in ACR caused by CycloZ alone or by the combination of dapagliflozin and CycloZ. Figure 2c This indicates the reduction in ACR caused by CycloZ alone or by the combined use of Losartan, Dapagliflozin, and CycloZ.

[0037] Figure 3a This indicates the reduction in ACR caused by CycloZ alone, sitagliptin alone, or sitagliptin + CycloZ combination administration, starting from the mild stage in KKAy mice, a model of diabetic nephropathy.

[0038] Figure 3b This indicates the changes in ACR caused by CycloZ alone, sitagliptin alone, or sitagliptin + CycloZ combination administration from the severe stage onwards in KKAy mice, a model of diabetic nephropathy.

[0039] Figure 4 This indicates the changes in ACR caused by administration of CycloZ alone, Losartan alone, Losartan + CycloZ combination, Losartan + Sitagliptin combination, or Losartan + Sitagliptin + CycloZ combination, starting from the severe stage in KKAy mice, a model of diabetic nephropathy.

[0040] Figure 5 This indicates the changes in ACR caused by CycloZ alone, metformin alone, or metformin + CycloZ combination administration from the severe stage onwards in KKAy mice, a model of diabetic nephropathy.

[0041] Figure 6 This indicates the reduction in ACR caused by CHP administration alone and in combination with zinc salts in KKAy mice, a model of diabetic nephropathy. Detailed Implementation

[0042] The present invention will now be described in more detail.

[0043] All technical terms used in this invention, unless otherwise defined, shall have the meaning commonly understood by one of ordinary skill in the art related to this invention. Furthermore, preferred methods or samples are described in this specification, but similar or equivalent methods are also included within the scope of this invention.

[0044] As mentioned above, existing therapeutic agents for diabetic nephropathy are practically unreliable in achieving effective therapeutic effects such as delaying or halting the progression of diabetic nephropathy. Therefore, the inventors have experimentally verified that the combination of zinc and cyclic histidine-proline not only demonstrates preventative, ameliorative, or therapeutic effects on diabetic nephropathy by reducing albuminuria, decreasing the albuminuria / creatinine ratio, or increasing glomerular filtration rate, but also significantly enhances the therapeutic efficacy of existing antihypertensive and / or antidiabetic agents prescribed to patients with diabetic nephropathy, thus exploring a solution to the aforementioned problems.

[0045] Therefore, a first aspect of the present invention relates to a pharmaceutical composition for the prevention or treatment of diabetic nephropathy, comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

[0046] Regarding the first aspect above, the present invention provides a method for the prevention, improvement or treatment of diabetic nephropathy, comprising administering to an individual in need of the method an effective amount of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0047] Regarding the first aspect above, the present invention also provides the use of a composition for the prevention, improvement or treatment of diabetic nephropathy, said composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0048] Regarding the first aspect above, the present invention also provides the use of a composition for preparing a medicament for preventing, improving or treating diabetic nephropathy, said composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0049] In this invention, the zinc salt may be a zinc salt selected from the group consisting of zinc ions, zinc metal, zinc chloride, zinc acetate, zinc gluconate, zinc stearate, zinc sulfate, zinc oxide, zinc picolinate, zinc orotate, and zinc citrate, but is not limited thereto.

[0050] In this invention, "cyclo-hispro (CHP)" refers to a naturally occurring cyclic dipeptide composed of histidine and proline, which are metabolites of thyrotropin-releasing hormone (TRH), or a physiologically active dipeptide synthesized in vivo during TRH metabolism and via de novo pathway, which is widely distributed throughout the brain, spinal cord, and gastrointestinal tract.

[0051] In this invention, the cyclic histidine-proline can be used synthetically or as a commercially available product. Alternatively, it can be used after purification from substances containing cyclic histidine-proline, such as prostate extracts and soybean hydrolysates.

[0052] The term "purified" indicates that it is a concentrated form compared to the cyclic histidine-proline form, which can be obtained from natural sources such as prostate extracts. Purified components can be obtained by concentration from their natural sources or by chemical synthesis.

[0053] In this invention, the zinc salt and cyclic histidine-proline can also be referred to as "Cyclo-Z," and the zinc salt and cyclic histidine-proline can be included in the composition of this invention as a complex or as individual components. In this case, the weight ratio of zinc salt to cyclic histidine-proline can be 1–10:1–5, preferably 1–5:1–2, but is not limited thereto. In this invention, the weight ratio relative to the zinc salt indicates the content of zinc cations, or the weight ratio of zinc components in the zinc salt.

[0054] Therefore, zinc salts and cyclic histidine-proline can be applied as a complex, or as individual components, applied simultaneously, alone, or sequentially.

[0055] In this invention, the pharmaceutical composition may further include an antihypertensive agent as another active ingredient.

[0056] Furthermore, in this invention, the composition applied to the individual may be administered in combination with an antihypertensive agent.

[0057] In this invention, the antihypertensive agent may be an ARB or an ACE inhibitor. The ARB and ACE are antihypertensive drugs that inhibit the renin-angiotensin-aldosterone system (RAAS). Specifically, ARB blocks angiotensin II from acting on angiotensin II receptor type 1 (AT1), while ACE inhibitors block the conversion of angiotensin I to angiotensin II by inhibiting ACE.

[0058] In this invention, the ARB may be any one selected from the group consisting of Losartan, Valsartan, Candesartan, Olmesartan, Telmisartan, Fimasartan, Irbesartan, Eprosartan, and Azilsartan, but is not limited thereto.

[0059] In this invention, the ACE inhibitor may be any one selected from the group consisting of captopril, zofinopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trundolapril, ciliapril, and fosinopril, but is not limited thereto.

[0060] In this invention, the weight ratio of the zinc salt and cyclic histidine-proline (Cyclo-Z):antihypertensive agent can be 1-2:1-4, wherein the concentration of the antihypertensive agent can be the same as or higher than that of the zinc salt and cyclic histidine-proline (Cyclo-Z), but is not limited thereto.

[0061] Angiotensin II (ARB) is the main end product of the renin-angiotensin system (RAS). By binding to AT1 receptors and through vasoconstriction, increased sodium reabsorption in renal tubules, water retention, decreased endothelial cell function, increased oxidative stress, tissue fibrosis, and increased collagen deposition, it is considered a central mediator in the development and progression of cardiovascular disease across multiple stages of the cardiovascular disease continuum. Angiotensin II receptor blockers (ARBs), by physiologically blocking AT1 receptors in the RAS, are currently the most widely used drugs for hypertension treatment due to their excellent antihypertensive effects and relatively few side effects. However, despite these advantages, the use of ARBs has also raised concerns about their safety due to the so-called ARB-myocardial infarction paradox—that they may actually increase the incidence of myocardial infarction—and the controversy surrounding their potential increase in cancer incidence.

[0062] In this invention, the pharmaceutical composition may further include an antidiabetic agent as another active ingredient.

[0063] Furthermore, in this invention, the composition applied to the individual may be administered in combination with an antidiabetic agent.

[0064] In this invention, the antidiabetic agent may be any one selected from the group consisting of SGLT2 inhibitors, DPP-4 inhibitors, and biguanide drugs.

[0065] In this invention, the SGLT2 inhibitor may be any one selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethyl acetate, and ertugliflozin, but is not limited thereto.

[0066] In this invention, the DPP-4 inhibitor may be any one selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin, but is not limited thereto.

[0067] The biguanide drug may be selected from any one of the group consisting of metformin, buformin, and phenformin, but is not limited thereto. In this invention, the weight ratio of the zinc salt and cyclic histidine-proline (Cyclo-Z):SGLT2 inhibitor can be 5-10:1-1.5, but is not limited thereto.

[0068] In this invention, the weight ratio of the zinc salt and cyclic histidine-proline (Cyclo-Z):DPP-4 inhibitor can be 1-3.5:1-2, wherein the concentration of the DPP-4 inhibitor can be the same as or lower than that of the zinc salt and cyclic histidine-proline (Cyclo-Z), but is not limited thereto.

[0069] In this invention, the weight ratio of the zinc salt and cyclic histidine-proline (Cyclo-Z):biguanide drug can be 1-2:10-16, but is not limited thereto.

[0070] In this invention, the pharmaceutical composition may further include the antihypertensive agent and the antidiabetic agent as another active ingredient. In this case, since the description of the antihypertensive agent and the antidiabetic agent is the same as above, their description is omitted.

[0071] Furthermore, in this invention, the composition applied to the individual may be used in combination with an antihypertensive agent and an antidiabetic agent. In this case, since the description of the antihypertensive agent and the antidiabetic agent is the same as that described above, their description is omitted.

[0072] In this invention, when the pharmaceutical composition comprises zinc salt and cyclic histidine-proline (Cyclo-Z), an antihypertensive agent and an SGLT2 inhibitor, the weight ratio of zinc salt and cyclic histidine-proline (Cyclo-Z): antihypertensive agent: SGLT2 inhibitor may be 5-10: 5-10: 0.5-1, wherein the concentration of the antihypertensive agent may be the same as or higher than that of the zinc salt and cyclic histidine-proline (Cyclo-Z), but is not limited thereto.

[0073] In this invention, when the pharmaceutical composition comprises a zinc salt and cyclic histidine-proline (Cyclo-Z), an antihypertensive agent, and a DPP-4 inhibitor, the weight ratio of the zinc salt and cyclic histidine-proline (Cyclo-Z): antihypertensive agent: DPP-4 inhibitor can be 5-10:5-10:2-5. The concentration of the antihypertensive agent can be the same as or higher than that of the zinc salt and cyclic histidine-proline (Cyclo-Z), and the concentration of the DPP-4 inhibitor can be lower than that of the zinc salt and cyclic histidine-proline (Cyclo-Z) and the antihypertensive agent, but is not limited thereto.

[0074] In this invention, the zinc salt and cyclic histidine-proline (Cyclo-Z) may be contained as a complex in clinical applications at a dose of 15 to 100 mg / kg, for example, at a dose of 38 mg / kg, 76 mg / kg, or 91 mg / kg per dosing unit, but not limited thereto. If the zinc salt and cyclic histidine-proline are contained as separate components, they may be contained within the above-mentioned dose range at a dose calculated according to the weight ratio of zinc salt to cyclic histidine-proline as defined in this invention.

[0075] In this invention, the SGLT2 inhibitor may be administered in clinical doses of 2 to 15 mg / kg, for example, in doses of 5 mg / kg or 10 mg / kg per dosing unit, but is not limited thereto.

[0076] In this invention, the DPP-4 inhibitor may be administered in doses of 10 to 120 mg / kg for clinical use. For example, each dosing unit may contain doses of 25 mg / kg, 50 mg / kg, or 100 mg / kg, but is not limited thereto.

[0077] In this invention, the biguanide drug may be administered in doses of 200 to 1500 mg / kg for clinical use. For example, each dosing unit may contain a dose of 500 mg / kg or 1000 mg / kg, but is not limited thereto.

[0078] Given the reported side effects of existing antihypertensive and antidiabetic agents, the inventors aim to explore a substance that can replace the combined use of these drugs, or significantly improve the therapeutic effect of these drugs on diabetic nephropathy, thereby preventing or reducing side effects caused by overdose or prolonged use of antihypertensive and / or antidiabetic agents. The results, such as... Figures 1a to 1dAs shown, in the KKay diabetic nephropathy mouse model, the combination of zinc salts and cyclic histidine-proline not only reduced albuminuria, but also significantly enhanced the albumin-reducing effects of these drugs when combined with losartan, a representative ARB drug, and / or dapagliflozin, a representative SGLT2 drug.

[0079] In addition, such as Figures 2a to 2c As shown, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclic histidine-proline (CycloZ) not only showed an effect of reducing ACR, but also significantly enhanced the ACR-reducing effect of these drugs when administered in combination with losartan and / or dapagliflozin.

[0080] In addition, as shown in Table 2, in the KKay diabetic nephropathy mouse model, the combination of zinc salts and cyclic histidine-proline, when administered in combination with losartan and / or dapagliflozin, further enhanced the glomerular filtration rate-increasing effects of these drugs.

[0081] In addition, such as Figure 3a and Figure 3b As shown, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclic histidine-proline, when administered in combination with sitagliptin, a representative drug of DPP-4 inhibitors, significantly improved the reduction of ACR compared with sitagliptin alone, regardless of the severity of diabetic nephropathy.

[0082] In addition, such as Figure 4 As shown, in the KKay diabetic nephropathy mouse model, the combination of cyclic histidine-proline, when administered in combination with losartan, a representative drug of ARBs, and sitagliptin, a representative drug of DPP-4 inhibitors, significantly improved the ACR reduction effect compared with the combination of losartan and sitagliptin alone.

[0083] In addition, such as Figure 5 As shown, in the KKay diabetic nephropathy mouse model, the combination of cyclic histidine-proline, when administered in combination with metformin, a representative biguanide drug, significantly improved the reduction of acute respiratory rate (ACR) compared with either cyclic histidine-proline alone or metformin alone.

[0084] Furthermore, such as Figure 6 As shown, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclic histidine-proline (CycloZ) significantly reduced the ACR value compared with cyclic histidine-proline alone, thus confirming the synergistic effect of the combined administration of zinc salt and cyclic histidine-proline.

[0085] Therefore, the pharmaceutical compositions of the present invention can exhibit one or more effects selected from the group consisting of i) reduced albuminuria; ii) reduced albuminuria / creatinine ratio; and iii) increased glomerular filtration rate.

[0086] In this invention, the diabetic nephropathy includes both type 1 and type 2 diabetic nephropathy, and regardless of the type of diabetes, it can exhibit the same effects as described in i) to iii).

[0087] The combination of zinc salts and cyclic histidine-proline is known to have a therapeutic effect by lowering blood glucose in type 2 diabetes, which is a non-insulin-dependent form of diabetes. However, it does not show significant therapeutic effects such as lowering blood glucose in type 1 diabetes, which is an insulin-dependent form of diabetes. Nevertheless, in diabetic nephropathy, regardless of the type of diabetes (i.e., including both type 1 and type 2), it can reduce albuminuria, lower the albuminuria / creatinine ratio, and increase glomerular filtration rate, thus demonstrating a preventive, ameliorative, or therapeutic effect on diabetic nephropathy through independent activity distinct from the therapeutic effect in type 2 diabetes.

[0088] In this invention, the pharmaceutical composition may be administered simultaneously, alone, or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0089] In this invention, the term "synergy effect" refers to an effect produced when the components are administered in combination, which is greater than the sum of the effects produced when each component is administered as a single component. This invention comprises compositions of zinc salts and cyclic histidine-proline or pharmaceutically acceptable salts thereof, which, when administered in combination with antihypertensive and / or antidiabetic agents, can enhance the preventive, ameliorative, or therapeutic effects on diabetic nephropathy.

[0090] The term "administered in combination" refers to the combined administration of compounds or ingredients to a patient. Combined administration means that, in order to achieve the desired therapeutic effect, the components may be administered at the same time, in any order, or sequentially at different times.

[0091] The term "prevention" as used in this invention refers to all actions that inhibit or delay the onset of a disease or condition. In this invention, it refers to delaying the onset of diabetic nephropathy or inhibiting its development.

[0092] The term “improvement” as used in this invention refers to all actions that improve or beneficially change the state of a disease or condition; in this invention, it refers to improving the symptoms of diabetic nephropathy.

[0093] The term "treatment" as used in this invention refers to all actions that delay, stop, or reverse the progression of a disease or condition; in this invention, it refers to reducing, alleviating, eliminating, or reversing the symptoms of diabetic nephropathy.

[0094] The term "pharmaceutically acceptable salt" as used in this invention refers to any organic or inorganic addition salt of cyclic histidine-proline, at an effective concentration that is relatively low in toxicity and harmless to the patient, and whose side effects do not diminish the beneficial efficacy of cyclic histidine-proline. These salts may use inorganic or organic acids as the free acid. Inorganic acids may include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc.; organic acids may include citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, edemaic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid, or malonic acid, etc. Furthermore, these salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, acid addition salts may include acetates, aspartates, benzoates, besylates, bicarbonates / carbonates, bisulfates / sulfates, borates, camphorates, citrates, edetates, ethanesulfonates, formates, fumarates, glucohepanoates, gluconates, glucuronates, hexafluorophosphates, hypothalamates, hydrochlorides / chlorides, hydrobromates / bromines, hydroiodates / iodides, hydroxyethanesulfonates, lactates, malates, maleates, malonates, and methanesulfonates. Salts, methyl sulfates, naphthalates, 2-naphthalenesulfonates, nicotinates, nitrates, orotates, oxalates, palmitates, pomaceates, phosphates / hydrogen phosphates / dihydrogen phosphates, saccharin salts, stearates, succinates, tartrates, p-toluenesulfonates, trifluoroacetates, as well as aluminum salts, arginine salts, benzyl sulfoxides, calcium salts, choline salts, diethylamine salts, ethanolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, oleylamine salts, potassium salts, sodium salts, tromethamine salts, zinc salts, etc.

[0095] The pharmaceutical compositions of the present invention may further comprise a pharmaceutically acceptable carrier. The pharmaceutical compositions comprising a pharmaceutically acceptable carrier may be in various dosage forms for oral or parenteral administration. In formulation, commonly used fillers, expanders, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients may be used. Solid dosage forms for oral administration may include tablets, pills, powders, granules, capsules, lozenges, etc., which may be prepared by mixing one or more compounds of the present invention with at least one or more excipients, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may be used. Liquid dosage forms for oral administration may include suspensions, oral solutions, emulsions, or syrups, and may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.

[0096] Formulations intended for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspension solvents, emulsions, lyophilized formulations, suppositories, etc. As non-aqueous solvents and suspension solvents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate can be used. As suppository bases, witepsol, polyethylene glycol, Tween 61, cocoa butter, lauryl ester, glycerin, gelatin, etc., can be used.

[0097] The pharmaceutical compositions of the present invention can be administered via any conventional route capable of reaching the target tissues or cells within an individual or sample. Administration may include systemic or local administration, and may include, but is not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, intranasal, intrapulmonary, and rectal administration. The dosage of the pharmaceutical composition may vary depending on the patient's age, weight, sex, administration method, health status, and disease severity.

[0098] In the methods of this invention, the term "individual" includes, but is not limited to, any animal (e.g., human, horse, pig, rabbit, dog, sheep, goat, non-human primate, cow, cat, guinea pig, or rodent). This term does not indicate a specific age or sex. Therefore, whether female or male, it is intended to include adult and newborn subjects, as well as fetuses. "Patient" refers to a subject suffering from a disease or disorder. The term "patient" includes both human and veterinary subjects.

[0099] In the method of the present invention, the description of the effects, route of administration, frequency of administration, dosage, etc. of the zinc salt and cyclic histidine-proline or their pharmaceutically acceptable salts applied to the individual is the same as that described above, and therefore is omitted.

[0100] In the method of this invention, when zinc salt and cyclic histidine-proline are applied in an effective amount, ideal preventive, ameliorative, or therapeutic effects on diabetic nephropathy can be provided. To achieve the desired effect, the combination of zinc salt and cyclic histidine-proline or their pharmaceutically acceptable salts can be administered once or repeatedly at regular intervals. In this case, the zinc salt and the cyclic histidine-proline can be administered simultaneously, separately, or sequentially. For example, zinc salt and cyclic histidine-proline or their pharmaceutically acceptable salts can be co-formulated into a unit-dose formulation for simultaneous administration, or administered simultaneously or sequentially as different formulations. Alternatively, compositions containing zinc salts and cyclic histidine-proline, when combined with antihypertensive agents and / or antidiabetic agents in an effective amount, can provide ideal preventive, ameliorative, or therapeutic effects for diabetic nephropathy. To achieve the desired effect, the combination of the present invention's composition containing zinc salts and cyclic histidine-proline or their pharmaceutically acceptable salts with antihypertensive agents and / or antidiabetic agents can be administered once or repeatedly at time intervals. In this case, the combination of the composition containing zinc salts and cyclic histidine-proline or their pharmaceutically acceptable salts with antihypertensive agents and / or antidiabetic agents can be administered simultaneously, separately, or sequentially. For example, the combination of the composition containing zinc salts and cyclic histidine-proline or their pharmaceutically acceptable salts with antihypertensive agents and / or antidiabetic agents can be simultaneously administered by co-formulating a unit-dose formulation of the combination, or administered simultaneously or sequentially as different formulations.

[0101] In the case of sequential administration, each active ingredient can be administered as an individual formulation at time intervals, and the order of administration can be determined by a physician or a person skilled in the art.

[0102] In addition, it can be used in combination with other methods for the prevention, improvement or treatment of diabetic nephropathy.

[0103] In the context of the present invention, descriptions of the effects of the composition comprising zinc salt and cyclic histidine-proline or a pharmaceutically acceptable salt thereof, as well as its route of administration, frequency of administration, dosage, etc., are omitted as they are the same as those described above.

[0104] According to the present invention, a composition comprising zinc salt and cyclic histidine-proline or a pharmaceutically acceptable salt thereof as an active ingredient can significantly improve the therapeutic effect of diabetic nephropathy when used in combination with ARB and / or SGLT2 inhibitors, and can therefore also be used as an adjuvant to enhance the therapeutic effect of these drugs on diabetic nephropathy.

[0105] Therefore, a second aspect of the present invention relates to a pharmaceutical composition for improving the preventive, ameliorative, or therapeutic effects of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

[0106] According to one embodiment of the present invention, a pharmaceutical composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient is provided for improving the therapeutic effect of ARB and / or SGLT2 inhibitors on diabetic nephropathy.

[0107] According to the present invention, the composition and effects of zinc salt and cyclic histidine-proline contained as active ingredients in pharmaceutical compositions for improving the therapeutic effect of ARB and / or SGLT2 inhibitors on diabetic nephropathy are described, and therefore are omitted as they are the same as those described above.

[0108] Furthermore, the types of antihypertensive and antidiabetic agents (specifically, ARBs and SGLT2s) that can exhibit synergistic effects with the zinc salt and cyclic histidine-proline combination are the same as those described above, and therefore their descriptions are omitted.

[0109] Regarding the second aspect, the present invention provides a method for improving the prevention, improvement, or treatment of diabetic nephropathy, comprising administering to an individual requiring the method an effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof. Here, the individual is the same as described in the first aspect above, and therefore its description is omitted. Alternatively, the individual may be a patient experiencing side effects due to overdose or prolonged use of antihypertensive and / or antidiabetic agents, or who is expected to experience such side effects. For example, the individual may be a diabetic nephropathy patient currently receiving antihypertensive and / or antidiabetic agents, preferably ARBs and / or SGLT2 inhibitors.

[0110] Regarding the second aspect, the present invention also provides the use of compositions comprising zinc salts; and cyclo-hispro or pharmaceutically acceptable salts thereof, for enhancing the preventive, ameliorative, or therapeutic effects of diabetic nephropathy.

[0111] Regarding the second aspect, the present invention also provides the use of compositions comprising zinc salts; and cyclo-hispro or pharmaceutically acceptable salts thereof, for the preparation of agents for enhancing the preventive, ameliorative, or therapeutic effects of diabetic nephropathy.

[0112] In this invention, the composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof can be used in combination with antihypertensive and / or antidiabetic agents to enhance the preventive, ameliorative, or therapeutic effects of antihypertensive and / or antidiabetic agents on diabetic nephropathy. In this case, the description of the antihypertensive and antidiabetic agents is the same as that described in the first aspect above, and therefore is omitted.

[0113] A third aspect of the present invention relates to a health food composition for the prevention or improvement of diabetic nephropathy, comprising zinc salt and cyclo-hispro or a food-acceptable salt thereof as active ingredients.

[0114] Regarding the third aspect, the present invention also provides the use of compositions comprising zinc salts; and cyclo-hispro or a food-acceptable salt thereof, for the manufacture of health foods for enhancing the prevention, improvement or treatment of diabetic nephropathy.

[0115] In this invention, the composition comprising zinc salt and cyclo-hispro or a food-acceptable salt thereof can be administered in combination to patients with diabetic nephropathy receiving antihypertensive and / or antidiabetic agents to enhance the preventive, ameliorative, or therapeutic effects of antihypertensive and / or antidiabetic agents on diabetic nephropathy. In this case, the description of the antihypertensive and antidiabetic agents is identical to that described in the first aspect above, and therefore is omitted.

[0116] According to one embodiment of the present invention, the health food composition can be used to improve the diabetic nephropathy improvement effect in patients with diabetic nephropathy who are receiving antihypertensive agents and / or antidiabetic agents (e.g., ARBs and / or SGLT2 inhibitors).

[0117] In the health food composition of the present invention, the description of the composition and effects of zinc salt and cyclic histidine-proline included as active ingredients is the same as described above, so the description is omitted.

[0118] In this invention, the term "food-acceptable salt" includes salts derived from food-acceptable organic acids, inorganic acids, or bases. Since the types of food-acceptable salts are the same as those described in the first aspect above for "pharmaceutical-acceptable salts," their description is omitted.

[0119] In this invention, the term "health food" includes both the meanings of "functional food" and "health food".

[0120] In this invention, the term "functional food" and "food for special health use (FoSHU)" are the same term, referring to foods that, in addition to providing nutrition, are processed to enable efficient biological regulatory functions and have high medical and therapeutic effects.

[0121] In this invention, the term "health food" refers to food that has a more positive effect on maintaining or improving health compared to ordinary food, while "health supplement food" refers to food intended to assist in health maintenance. In some cases, the terms functional food, health food, and health supplement food may be used interchangeably. The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, and pills to obtain beneficial effects for the prevention or improvement of diabetic nephropathy.

[0122] As a specific example of such functional foods, the zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can be used to manufacture processed foods that improve the shelf life while giving full play to the characteristics of agricultural, livestock or aquatic products. The health food composition of the present invention can also be prepared in the form of nutritional supplements, food additives, and feed, and is intended for human consumption or for animals including livestock.

[0123] The above-described food compositions can be prepared in various forms according to conventional methods known in the art. As general foods, the zinc salt and cyclic histidine-proline or their food-grade salts of the present invention can be added to, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled foods, jams, marmalade, etc.), fish, meat and their processed foods (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon noodles, soba noodles, instant noodles, pasta, macaroni, etc.), fruit juices, various beverages, biscuits, syrups, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, ready-to-eat foods, frozen foods, and various seasonings (e.g., miso, soy sauce, condiments, etc.).

[0124] In addition, as a nutritional supplement, the zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can be added to, but are not limited to, capsules, tablets, pills, etc.

[0125] Furthermore, as health food products, the zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can be prepared into tea, juice, and beverage forms, and consumed (health drinks) or ingested in liquid, granulated, encapsulated, or powdered form, but are not limited thereto. In addition, for use as a food additive, the zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can be prepared as powders or concentrated liquids. Furthermore, the zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can also be mixed with known active ingredients known to be effective in the prevention or improvement of diabetic nephropathy to prepare a composition.

[0126] When the food composition of the present invention is used as a health beverage composition, the health beverage composition may, like ordinary beverages, further contain various flavoring agents or natural carbohydrates and other additional ingredients. The aforementioned natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; or sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners may be natural sweeteners such as sweetened protein and stevia extract; or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates is typically about 0.01–0.04 g per 100 mL of the composition of the present invention, preferably about 0.02–0.03 g.

[0127] The zinc salt and cyclic histidine-proline or their food-acceptable salts of the present invention can be included as active ingredients in health food compositions for the prevention or improvement of diabetic nephropathy, in an effective amount sufficient to achieve the aforementioned preventive or improving effects, preferably 0.01 to 100% by weight relative to the total weight of the whole composition, but not limited thereto. The health food compositions of the present invention can be prepared by mixing zinc salt and cyclic histidine-proline or their food-acceptable salts with known active ingredients known to be effective in the prevention or improvement of diabetic nephropathy.

[0128] In addition to the above, the health food products of the present invention may also contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, pectate, alginic acid, alginate, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonation agents, etc. Furthermore, the health food products of the present invention may also contain fruit pulp used in the manufacture of natural fruit juices, fruit juice beverages, or vegetable beverages. These ingredients may be used alone or in combination. The proportions of these additives are not particularly important, but are generally selected from a range of 0.01 to 0.1 parts by weight relative to 100 parts by weight of the composition of the present invention.

[0129] Implementing the embodiments of the present invention The present invention will be described in more detail below through embodiments. These embodiments are for illustrative purposes only, and those skilled in the art should understand that the scope of the present invention should not be construed as limited to these embodiments.

[0130]

Example 1

[0131] It has been reported that the KKAy animal model exhibits 10 to 50 times higher levels of albuminuria than the db / db animal model, and more severe mesangial matrix expansion. Five-week-old KKAy mice purchased from CleaJapan were pre-fed for one week and then grouped as shown in Table 1. Losartan was dissolved in drinking water, with each animal receiving 20 mg / kg daily; dapagliflozin (2 mg / kg), cyclozil (15 mg / kg), or a combination thereof were administered orally daily. The normal control group and the negative control group received the same dose of distilled water orally daily.

[0132] Table 1

[0133] 1-2. Urine albumin assay At week 16 of administration, 24-hour urine was collected using a metabolic cage and centrifuged at 2,000 rpm for 10 minutes to remove suspended matter. Albumin was measured using urine diluted to 1 / 10 with distilled water using a DCA vantage analyzer (Siemens) and a DCA® Microalbumin / Creatinine Urine Test card (Siemens). The results are as follows Figure 1a As shown, the albuminuria (mg / dL) level was 51.4 in the negative control group, 39.3 in the losartan monotherapy group (group 1), 45.4 in the cyclozinc monotherapy group (group 3), and 11.1 in the losartan + cyclozinc combination therapy group (group 4). Compared with the negative control group, each drug group showed a reduction in albuminuria of 23.5%, 11.7%, and 78.4%, respectively. Based on these results, it is confirmed that the losartan + cyclozinc combination therapy has a synergistic effect in reducing albuminuria.

[0134] In addition, such as Figure 1b As shown, the albuminuria (mg / dL) level was 51.4 in the negative control group, 31.1 in the dapagliflozin monotherapy group (group 2), 45.4 in the cyclozinc monotherapy group (group 3), and 22.4 in the dapagliflozin + cyclozinc combination group (group 5). Compared with the negative control group, each drug group showed a reduction of 39.5%, 11.7%, and 56.4%, respectively. Based on these results, it is confirmed that the dapagliflozin + cyclozinc combination therapy has a synergistic effect in reducing albuminuria.

[0135] In addition, such as Figure 1c As shown, albuminuria (mg / dL) was 51.4 in the negative control group, 45.4 in the CycloZ monotherapy group (group 3), 6.7 in the Losartan + Dapagliflozin combination group (group 6), and 2.2 in the Losartan + Dapagliflozin + CycloZ combination group (group 7). These drug administration groups showed reductions of 11.7%, 87%, and 95.7% compared to the negative control group, respectively. Based on these results, it is confirmed that the Losartan + Dapagliflozin + CycloZ combination therapy exhibits a synergistic effect in reducing albuminuria.

[0136] Finally, as Figure 1d The results shown represent an overall comparison of albuminuria measurement results in groups 1 to 7. The albuminuria (mg / dL) level was 51.4 in the negative control group, 39.3 in the losartan monotherapy group (group 1), 31.1 in the dapagliflozin monotherapy group (group 2), 45.4 in the cyclozinc monotherapy group (group 3), 11.1 in the losartan + cyclozinc combination group (group 4), 22.4 in the dapagliflozin + cyclozinc combination group (group 5), 6.7 in the losartan + dapagliflozin combination group (group 6), and 2.2 in the losartan + dapagliflozin + cyclozinc combination group (group 7). Compared with the negative control group, each drug administration group showed a reduction of 23.5%, 39.5%, 11.7%, 56.4%, 78.4%, 87%, and 95.7%, respectively. In particular, the combination of losartan, dapagliflozin, and cyclozinc (CycloZ) showed a significantly better reduction in albuminuria compared to the individual drug groups or the combination of the two drugs, thus confirming that the synergistic effect of the three combination drugs was the best. 1-3. Confirmation of albuminuria / creatinine ratio Creatinine was measured using undiluted urine via a mouse creatinine assay kit (Crystalchem). The albumin (mg / dL) value was divided by the creatinine (g / dL) value to derive the ACR (mg / g) value. The results are as follows Figure 2a As shown, the ACR value of the losartan + cyclozan (CycloZ) combination group (group 4) was further reduced compared with the losartan or cyclozan alone groups (group 1 or group 3), confirming that the combination administration produced a synergistic effect.

[0137] In addition, such as Figure 2b As shown, the ACR value of the dapagliflozin + cyclozil (CycloZ) combination group (group 5) was further reduced compared with the dapagliflozin or cyclozil monotherapy groups (group 2 or group 3), confirming that the combination therapy produced a synergistic effect.

[0138] Finally, as Figure 2c As shown, the ACR value of the losartan + dapagliflozin + cyclozinc (CycloZ) combination group (group 7) was significantly lower than that of the losartan, dapagliflozin or cyclozinc alone groups (groups 1 to 3), and was also further lower than that of the losartan + dapagliflozin combination group.

[0139]

Example 2

[0140] The results are shown in Table 2. The glomerular filtration rate (GFR) of the losartan + dapagliflozin + cyclozinc combination group (group 7) was significantly increased compared with the negative control group, and was also further increased compared with the losartan + cyclozinc combination group (group 4) and the dapagliflozin + cyclozinc (CycloZ) combination group (group 5).

[0141] Table 2

[0142]

Example 3

[0143] Table 3

[0144] 3-2. Confirmation of albuminuria / creatinine ratio in mild cases At week 11 of drug administration, 24-hour urine was collected using a metabolic cage and centrifuged at 1,000 rpm for 5 minutes to remove suspended solids. Albumin was measured using urine diluted to half with distilled water using a DCA vantage analyzer (Siemens) and a DCA® Microalbumin / Creatinine Urine Test card (Siemens). Creatinine was measured using undiluted urine using a mouse creatinine assay kit (Crystalchem). The ACR (mg / g) value was derived by dividing the measured albumin (mg / dL) value by the creatinine (g / dL) value. The results are as follows Figure 3a As shown, the ACR (mg / g) value was 6,379 mg / g in the negative control group, 4,042 mg / g in the CycloZ monotherapy group, 4,852 mg / g in the sitagliptin monotherapy group, and 2,378 mg / g in the sitagliptin + CycloZ combination therapy group. These values ​​represent reductions of approximately 37%, 24%, and 63% respectively compared to the control group. Since the sitagliptin + CycloZ combination therapy group exhibited a significantly improved ACR reduction compared to the other monotherapy groups, it is confirmed that it demonstrates a synergistic effect induced by CycloZ.

[0145] 3-3. Medications administered in severe cases After feeding 5-week-old KKAy mice from Examples 1-1 for 6 weeks, 24-hour urine was collected, and ACR was measured using the same method as in Examples 3-2. Subsequently, to ensure consistent mean ACR values, the mice were randomly assigned to groups as shown in Table 4, and were orally administered the drugs daily as shown in Table 4 starting at 12 weeks of age for 11 weeks.

[0146] Table 4

[0147] 3-4. Confirmation of albuminuria / creatinine ratio in severe cases At week 11 of administration, 24-hour urine was collected using a metabolic cage, and ACR was determined using the same method as in Examples 3-2.

[0148] The results are as follows Figure 3bAs shown, the ACR (mg / g) value was 4,450 mg / g in the negative control group, 4,701 mg / g in the CycloZ monotherapy group, 2,301 mg / g in the sitagliptin monotherapy group, and 1,591 mg / g in the sitagliptin + CycloZ combination therapy group. When administration began at the onset of severe illness, CycloZ monotherapy did not show a significant effect on ACR improvement, but the sitagliptin monotherapy group showed a 48% reduction compared to the negative control group, while the sitagliptin + CycloZ combination therapy group showed a 31% reduction compared to the sitagliptin monotherapy group and a 64% reduction compared to the negative control group. This confirms that the sitagliptin + CycloZ combination therapy has a synergistic effect on improving diabetic nephropathy in the severe stage.

[0149]

Example 4

[0150] Table 5

[0151] 4-2. Confirmation of albuminuria / creatinine ratio in severe cases At week 9 of administration, 24-hour urine was collected using a metabolic cage, and ACR was determined using the same method as in Examples 3-2.

[0152] The results are as follows Figure 4As shown, the ACR (mg / g) value was 7,118 mg / g in the negative control group, 6,639 mg / g in the CycloZ monotherapy group, 7,158 mg / g in the losartan monotherapy group, and 4,125 mg / g in the losartan + CycloZ combination therapy group. When administration began from the severe stage, neither CycloZ nor losartan monotherapy showed a significant effect on ACR improvement, but the losartan + CycloZ combination therapy group showed a 42% reduction compared to the negative control group. This confirms that the losartan + CycloZ combination therapy has a synergistic effect on improving diabetic nephropathy in the severe stage.

[0153] In addition, such as Figure 4 As shown, the ACR value of the losartan + sitagliptin combination group was 2,768 mg / g, while that of the losartan + sitagliptin + cyclozinc (CycloZ) combination group was 2,025 mg / g. These values ​​were 61.1% and 71.6% lower than those of the negative control group, respectively. Furthermore, since the ACR value of the losartan + sitagliptin + cyclozinc (CycloZ) combination group was further lower than that of the losartan + sitagliptin combination group, it was confirmed that it exhibited a synergistic effect brought about by CycloZ.

[0154]

Example 5

[0155] Table 6

[0156] 5-2. Confirmation of albuminuria / creatinine ratio in severe cases At week 12 of administration, 24-hour urine was collected using a metabolic cage, and ACR was determined using the same method as in Examples 3-2.

[0157] The results are as follows Figure 5As shown, the ACR (mg / g) value was 2,369 mg / g in the negative control group, 2,792 mg / g in the CycloZ monotherapy group, 2,952 mg / g in the metformin monotherapy group, and 1,117 mg / g in the metformin + CycloZ combination therapy group. When administration began at the onset of severe illness, neither CycloZ nor metformin monotherapy showed a significant effect on ACR improvement, but the metformin + CycloZ combination therapy group showed a 53% reduction compared to the negative control group. This confirms that the metformin + CycloZ combination therapy has a synergistic effect on improving severe diabetic nephropathy.

[0158]

Example 6

[0159] Table 7

[0160] Creatinine was measured using undiluted urine via a mouse creatinine assay kit (Crystalchem). The albumin (mg / dL) value was divided by the creatinine (g / dL) value to derive the ACR (mg / g) value. Results are as follows... Figure 6 As shown, the ACR value of the CycloZ group (group 3) was significantly lower than that of the CHP monotherapy group (group 1 or group 2), confirming that the combination of CHP and Zn showed a synergistic effect compared to CHP monotherapy.

[0161] The foregoing has described specific parts of the present invention in detail. However, it should be understood by those skilled in the art that these specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Therefore, the essential scope of the invention should be defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for the prevention or treatment of diabetic nephropathy, comprising: Zinc salts; and Cyclic histidine-proline or its pharmaceutically acceptable salts are used as the active ingredient.

2. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 1, comprising: Antihypertensive agents selected from the group consisting of angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors were used as additional active ingredients.

3. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 1, comprising: Antidiabetic agents from the group consisting of sodium-glucose cotransporter 2 inhibitors, dipeptidyl peptidase-4 inhibitors, and biguanide drugs were selected as additional active ingredients.

4. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 1, comprising: Antihypertensive agents selected from the group consisting of angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors; and Antidiabetic agents from the group consisting of sodium-glucose cotransporter 2 inhibitors, dipeptidyl peptidase-4 inhibitors, and biguanide drugs were selected as additional active ingredients.

5. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 2 or 4, wherein, The angiotensin II receptor blocker is selected from any one of the following groups: losartan, valsartan, candesartan, olmesartan, telmisartan, femasartan, irbesartan, eprosartan, and azilsartan. The angiotensin-converting enzyme inhibitor is selected from any one of the following groups: captopril, zofinopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trundolapril, ciliapril, and fosinopril.

6. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 3 or 4, wherein, The sodium-glucose cotransporter-2 inhibitor is selected from any one of the following groups: dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin. The dipeptidylpeptidase-4 inhibitor is selected from any one of the following groups: sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin. The biguanide drug is selected from any one of the group consisting of metformin, buformin, and phenformin.

7. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 1, wherein, The pharmaceutical composition exhibits one or more effects selected from the group consisting of i) to iii): i) Decreased albuminuria; ii) Decreased albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

8. The pharmaceutical composition for the prevention or treatment of diabetic nephropathy according to claim 1, wherein, The pharmaceutical composition may be administered simultaneously, alone, or sequentially with antihypertensive agents and / or antidiabetic agents. The antihypertensive agent is selected from any one of the following groups: angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors. The antidiabetic agent is selected from any one of the following groups: sodium-glucose cotransporter 2 inhibitors, dipeptidyl peptidase-4 inhibitors, and biguanide drugs.

9. A pharmaceutical composition for enhancing the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, comprising: Zinc salts; and Cyclic histidine-proline or its pharmaceutically acceptable salts are used as the active ingredient.

10. The pharmaceutical composition according to claim 9 for enhancing the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, wherein, The antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, femasartan, irbesartan, eprosartan, and azilsartan; or Angiotensin-converting enzyme inhibitors selected from the group consisting of captopril, zofinopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, ciliapril, and fosinopril.

11. The pharmaceutical composition according to claim 9 for enhancing the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, wherein, The antidiabetic agent is a sodium-glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin. Select a dipeptidyl peptidase-4 inhibitor from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin; or Biguanide drugs selected from the group consisting of metformin, buformin, and phenformin.

12. The pharmaceutical composition according to claim 9 for enhancing the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, wherein, The pharmaceutical composition exhibits one or more effects selected from the group consisting of i) to iii): i) Decreased albuminuria; ii) Decreased albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

13. The pharmaceutical composition according to claim 9 for enhancing the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, wherein, The pharmaceutical composition is administered simultaneously, alone, or sequentially with an antihypertensive agent and / or an antidiabetic agent, wherein the antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, femasartan, irbesartan, eprosartan, and azilsartan; or an angiotensin-converting enzyme inhibitor selected from the group consisting of captopril, zofinopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cirazolipril, and fosinopril. The antidiabetic agent is a sodium-glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin; sitagliptin is also selected. The drug is selected from the group consisting of dipeptidyl peptidase-4 inhibitors of liptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin; or biguanide drugs selected from the group consisting of metformin, buformin, and phenformin.

14. A health food composition for preventing or improving diabetic nephropathy, comprising: Zinc salts; and Cyclic histidine-proline or its food-acceptable salts are used as the active ingredient.

15. The health food composition for preventing or improving diabetic nephropathy according to claim 14, wherein, The health food composition is used to improve the therapeutic effect of diabetic nephropathy in patients with diabetic nephropathy who are receiving antihypertensive and / or antidiabetic agents. The antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, femasartan, irbesartan, eprosartan, and azilsartan; or an angiotensin-converting enzyme inhibitor selected from the group consisting of captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cirazopril, and fosinopril. The antidiabetic agent is a sodium-glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin; sitagliptin is also selected. The drug is selected from the group consisting of dipeptidyl peptidase-4 inhibitors of liptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin; or biguanide drugs selected from the group consisting of metformin, buformin, and phenformin.

16. The health food composition for preventing or improving diabetic nephropathy according to claim 14, wherein, The health food composition exhibits one or more effects selected from the group consisting of i) to iii): i) Decreased albuminuria; ii) Decreased albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

17. The health food composition for preventing or improving diabetic nephropathy according to claim 14, wherein, The health food composition may be used simultaneously, alone, or sequentially with antihypertensive agents and / or antidiabetic agents. The antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, femasartan, irbesartan, eprosartan, and azilsartan; or an angiotensin-converting enzyme inhibitor selected from the group consisting of captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cirazopril, and fosinopril. The antidiabetic agent is a sodium-glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin ethacrylate, and ertugliflozin; sitagliptin is also selected. The drug is selected from the group consisting of dipeptidyl peptidase-4 inhibitors of liptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, tenegliptin, anagliptin, and evogliptin; or biguanide drugs selected from the group consisting of metformin, buformin, and phenformin.

18. A method for preventing, improving, or treating diabetic nephropathy, comprising the following steps: The effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof is administered to the individual in need.

19. A method for improving the therapeutic effect of antihypertensive agents and / or antidiabetic agents on diabetic nephropathy, comprising the following steps: The effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof is administered to the individual in need.

20. Use of a composition for preparing a medicament for the prevention, improvement, or treatment of diabetic nephropathy, said composition comprising: Zinc salts; and Cyclic histidine-proline or its pharmaceutically acceptable salt.

21. Use of a composition for preparing a pharmaceutical agent for the prevention, improvement, or treatment of diabetic nephropathy by an antihypertensive agent and / or an antidiabetic agent, said composition comprising: Zinc salts; and Cyclic histidine-proline or its pharmaceutically acceptable salt.

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