Use of composition for preventing, alleviating or treating diabetic kidney disease comprising zinc salt and cyclo-hispro as active ingredients

Abstract

The present invention relates to a use of a composition for preventing, alleviating, or treating diabetic kidney disease, the composition comprising zinc salt and cyclo-hispro as active ingredients. More specifically, the present invention provides: a composition for preventing, alleviating, or treating diabetic kidney disease, the composition comprising zinc salt and cyclo-hispro as active ingredients, wherein the composition, when administered alone or in combination with an antihypertensive agent and / or antidiabetic agent, exhibits the effect of preventing, alleviating, or treating diabetic kidney disease through the effect of reducing albuminuria, reducing the albuminuria / creatinine ratio, or increasing the glomerular filtration rate; and a method for preventing, alleviating, or treating diabetic kidney disease using same.

Description

Use of a composition comprising zinc salt and cyclo-hispro as active ingredients for the prevention, improvement or treatment of diabetic nephropathy

[0001] This application claims priority to Republic of Korea Patent Application No. 10-2023-0153151, filed on November 7, 2023, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention relates to the use of a composition comprising a zinc salt and cyclo-hispro as active ingredients for the prevention, improvement or treatment of diabetic nephropathy, and more particularly, to a composition comprising a zinc salt and cyclo-hispro as active ingredients, which exhibits a prevention, improvement or treatment effect of diabetic nephropathy through the effect of reducing albuminuria, reducing the albuminuria / creatinine ratio or increasing the glomerular filtration rate when administered alone or in combination with an antihypertensive and / or antidiabetic agent, and to a method for preventing, improving or treating diabetic nephropathy using the same.

[0003] Diabetic kidney disease (DKD) is a complication of diabetes, affecting approximately 40% of diabetic patients (Non-patent Document 1). While the etiology of DKD remains unclear, it is believed to be caused by damage to blood vessels, cells, and tissues within the kidneys due to persistent hyperglycemia, hypertension, obesity, a high-salt diet, and genetic factors (Non-patent Document 2). Kidney damage caused by DKD causes proteinuria (albuminuria) and reduces the glomerular filtration rate (GFR), impairing the kidney's vital filtration and reabsorption functions and becoming a major cause of end-stage renal failure.

[0004] Currently, angiotensin receptor blockers (ARBs), such as losartan, which regulate blood pressure, and sodium glucose cotransporter 2 (SGLT2) inhibitors, such as dapagliflozin, which regulate blood sugar, are used for the prevention and treatment of DKD. However, losartan, which is classified as an angiotensin II receptor blocker, can cause common side effects such as indigestion, diarrhea, and abdominal pain. Dapagliflozin, which is the SGLT2 inhibitor with the most clinical results, is commonly reported to cause side effects such as headache, diarrhea, back pain, bronchitis, pharyngitis, and upper respiratory tract infection. Furthermore, routine treatments for blood pressure and blood sugar control alone are unlikely to be effective in slowing or halting the progression of DKD. Therefore, strategic drug development that enhances the level of therapeutic efficacy through new drugs or synergistic effects through combinations with existing drugs is necessary.

[0005] Meanwhile, Patent Document 1 discloses a composition containing zinc ions and cyclo-hispro (CHP) as a composition useful for alleviating diabetic symptoms in mammals, but no example of combining the composition with an antihypertensive agent and / or an antidiabetic agent for the purpose of preventing, improving, or treating DKD is disclosed.

[0006] Against this backdrop, the present inventors have confirmed that the combination of zinc and cyclo-hispro not only exhibits the effect of preventing, improving, or treating diabetic nephropathy through the effect of reducing albuminuria, reducing the albuminuria / creatinine ratio, or increasing the glomerular filtration rate, but also significantly increases the diabetic nephropathy treatment effect of antihypertensive and / or antidiabetic drugs prescribed to patients with diabetic nephropathy when administered in combination with these drugs, thereby completing the present invention.

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] (Patent Document 1) Republic of Korea Publication No. 10-2001-0022786

[0010] [Non-patent literature]

[0011] (Non-patent literature 1) Clin J Am Soc Nephrol : CJASN., 12 (2017), pp. 2032-204

[0012] (Non-patent literature 2) Int J Mol Sci. 2020 Mar 23;21(6):2218

[0013] Accordingly, the present invention aims to provide a composition for preventing, improving or treating diabetic nephropathy, comprising a combination of zinc and cyclo-hispro.

[0014] In addition, the present invention aims to provide a composition for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy, comprising a combination of zinc and cyclo-hispro.

[0015] In addition, the present invention aims to provide a method for preventing, improving or treating diabetic nephropathy using a combination of zinc and cyclo-hispro.

[0016] Furthermore, the present invention aims to provide a method for improving the effect of an antihypertensive and / or antidiabetic agent in preventing, improving or treating diabetic nephropathy using a combination of zinc and cyclo-hispro.

[0017] Additionally, the present invention aims to provide a use of a composition comprising a combination of zinc and cyclo-hispro for the manufacture of a medicament for preventing, improving or treating diabetic nephropathy.

[0018] In addition, the present invention aims to provide a use of a composition comprising a combination of zinc and cyclo-hispro for the manufacture of a drug for enhancing the effect of an antihypertensive and / or antidiabetic agent in preventing, improving or treating diabetic nephropathy.

[0019] To solve the above-described problem, the present invention provides a pharmaceutical composition for preventing or treating diabetic nephropathy, comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients.

[0020] The present invention also provides a pharmaceutical composition for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy, the pharmaceutical composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients.

[0021] The present invention also provides a health functional food composition for preventing or improving diabetic nephropathy, comprising zinc salt and cyclo-hispro or a food-related acceptable salt thereof as active ingredients.

[0022] In the present invention, the pharmaceutical composition may contain, as an additional active ingredient, an antihypertensive agent selected from the group consisting of an angiotensin II receptor blocker (ARB) and an angiotensin-converting-enzyme inhibitor (ACE inhibitor).

[0023] In the present invention, the pharmaceutical composition may contain, as an additional active ingredient, an antidiabetic agent selected from the group consisting of a sodium glucose cotransporter 2 inhibitor (SGLT2 inhibitor), a dipeptidyl peptidase-4 inhibitor (DPP-4 inhibitor), and a biguanide drug.

[0024] In the present invention, the pharmaceutical composition may further comprise an antihypertensive agent selected from the group consisting of ARB and ACE inhibitors and an antidiabetic agent selected from the group consisting of SGLT2 inhibitors, DPP-4 inhibitors and biguanide drugs as an additional active ingredient.

[0025] In the present 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 any one selected from the group consisting of Captopril, Zofenopril, Enalapril, Ramipril, Quinapril, Perindopril, Lisinopril, Benazepril, Imidapril, Trandolapril, It may be any one selected from the group consisting of cilazapril and fosinopril.

[0026] In the present invention, the SGLT2 inhibitor may be any one selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin, and the DPP-4 inhibitor may be any one selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, It may be any one selected from the group consisting of anagliptin and evogliptin, and the biguanide drug may be any one selected from the group consisting of metformin, buformin, and phenformin.

[0027] In the present invention, the composition may exhibit at least one effect selected from the group consisting of i) reducing albuminuria; ii) reducing albuminuria / creatinine ratio; and iii) increasing glomerular filtration rate.

[0028] In the present invention, the composition may be administered simultaneously, separately or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0029] In the present invention, the health functional food composition may be for improving the diabetic nephropathy improvement effect in a diabetic nephropathy patient receiving the antihypertensive agent and / or the antidiabetic agent.

[0030] Additionally, the present invention provides a method for preventing, improving or treating diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0031] In addition, the present invention provides a method for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0032] Furthermore, the present invention provides a use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing, improving or treating diabetic nephropathy.

[0033] In addition, the present invention provides a use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing, improving, or enhancing the effect of treating diabetic nephropathy by an antihypertensive and / or antidiabetic agent.

[0034] In the present invention, the composition may contain an antihypertensive agent selected from the group consisting of an angiotensin II receptor blocker (ARB) and an angiotensin-converting-enzyme inhibitor (ACE inhibitor) as an additional active ingredient, or may be administered in combination with the antihypertensive agent.

[0035] In the present invention, the composition may further comprise an antidiabetic agent selected from the group consisting of a sodium glucose cotransporter 2 inhibitor (SGLT2 inhibitor), a dipeptidyl peptidase-4 inhibitor (DPP-4 inhibitor), and a biguanide drug as an additional active ingredient, or may be administered in combination with the antidiabetic agent.

[0036] In the present invention, the composition may further comprise an antihypertensive agent selected from the group consisting of ARB and ACE inhibitors and an antidiabetic agent selected from the group consisting of SGLT2 inhibitors, DPP-4 inhibitors and biguanide drugs as an additional active ingredient, or may be administered in combination with the antihypertensive agent and the antidiabetic agent.

[0037] In the present 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 any one selected from the group consisting of Captopril, Zofenopril, Enalapril, Ramipril, Quinapril, Perindopril, Lisinopril, Benazepril, Imidapril, Trandolapril, It may be any one selected from the group consisting of cilazapril and fosinopril.

[0038] In the present invention, the SGLT2 inhibitor may be any one selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin, and the DPP-4 inhibitor may be any one selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, It may be any one selected from the group consisting of anagliptin and evogliptin, and the biguanide drug may be any one selected from the group consisting of metformin, buformin, and phenformin.

[0039] In the present invention, the composition may exhibit at least one effect selected from the group consisting of i) reducing albuminuria; ii) reducing albuminuria / creatinine ratio; and iii) increasing glomerular filtration rate.

[0040] In the present invention, the composition may be administered simultaneously, separately or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0041] The composition comprising a zinc salt and cyclo-hispro according to the present invention exhibits a preventive, ameliorating, or therapeutic effect on diabetic nephropathy through the effects of reducing albuminuria, reducing the albuminuria / creatinine ratio, or increasing the glomerular filtration rate when the combination is administered alone, and thus can be utilized as a therapeutic agent and health functional food for the prevention, amelioration, or treatment of diabetic nephropathy. In addition, when administered in combination with antihypertensive and / or antidiabetic drugs prescribed to patients with diabetic nephropathy, the diabetic nephropathy therapeutic effect of these drugs is significantly increased, and thus, the composition is useful as an adjuvant for patients receiving the above drugs, and can also contribute to reducing side effects caused by chronic administration of the above drugs.

[0042] Figures 1a to 1d are the results of confirming the albuminuria reduction effect according to the administration of CycloZ or various drug combinations in KKAy mice, a diabetic nephropathy model. Figure 1a shows the albuminuria reduction effect by administering CycloZ alone or a combination of losartan and CycloZ, Figure 1b shows the albuminuria reduction effect by administering CycloZ alone or a combination of dapagliflozin and CycloZ, Figure 1c shows the albuminuria reduction effect by administering CycloZ alone or a combination of losartan, dapagliflozin and CycloZ, and Figure 1d shows the results of Figures 1a to 1c compared in one graph.

[0043] Figures 2a to 2c are the results of confirming the effect of reducing the urine albumin to creatinine ratio (ACR) according to the administration of CycloZ or various drug combinations in KKAy mice, a diabetic nephropathy model. Figure 2a shows the ACR reducing effect by administering CycloZ alone or a combination of losartan and CycloZ, Figure 2b shows the ACR reducing effect by administering CycloZ alone or a combination of dapagliflozin and CycloZ, and Figure 2c shows the ACR reducing effect by administering CycloZ alone or a combination of losartan, dapagliflozin, and CycloZ.

[0044] Figure 3a shows the ACR reduction effect by administration of CycloZ alone, sitagliptin alone, or sitagliptin + CycloZ combination from the mild stage in KKAy mice, a diabetic nephropathy model.

[0045] Figure 3b shows the changes in ACR caused by administration of CycloZ alone, sitagliptin alone, or sitagliptin + CycloZ combination from the severe stage in KKAy mice, a diabetic nephropathy model.

[0046] Figure 4 shows the changes in ACR due to administration of CycloZ alone, losartan alone, losartan + CycloZ combination, losartan + sitagliptin combination, or losartan + sitagliptin + CycloZ combination from the severe stage in KKAy mice, a diabetic nephropathy model.

[0047] Figure 5 shows the changes in ACR caused by administration of CycloZ alone, metformin alone, or a combination of metformin and CycloZ from the severe stage of KKAy mice, a diabetic nephropathy model.

[0048] Figure 6 shows the ACR reduction effect of CHP alone and a combination of zinc salt and CHP in KKAy mice, a diabetic nephropathy model.

[0049] Hereinafter, the present invention will be described in more detail.

[0050] All technical terms used in this invention, unless otherwise defined, have the same meaning as commonly understood by those skilled in the art. While preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of this invention.

[0051] As described above, it is difficult to expect effective treatment, such as slowing or stopping the progression of diabetic nephropathy, with existing diabetic nephropathy treatments. Therefore, the present inventors have experimentally verified that the combination of zinc and cyclo-hispro not only has the effect of preventing, improving, or treating diabetic nephropathy through the effects of reducing albuminuria, reducing the albuminuria / creatinine ratio, or increasing the glomerular filtration rate, but also significantly increases the diabetic nephropathy treatment effect of antihypertensive and / or antidiabetic drugs prescribed to patients with diabetic nephropathy when administered together, thereby seeking a solution to the above-mentioned problem.

[0052] Accordingly, the present invention relates to a pharmaceutical composition for preventing or treating diabetic nephropathy, comprising, in a first aspect, a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients.

[0053] In relation to the first aspect, the present invention provides a method for preventing, improving or treating diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof.

[0054] In connection with the first aspect, the present invention also provides the use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof, for use in the prevention, improvement or treatment of diabetic nephropathy.

[0055] In connection with the first aspect, the present invention also provides the use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for preventing, improving or treating diabetic nephropathy.

[0056] In the present invention, the zinc salt may be a zinc salt selected from the group consisting of zinc ion, zinc metal, or zinc chloride, zinc acetate, zinc gluconate, zinc stearate, zinc sulfate, zinc oxide, zinc picolinate, zinc orotate, and zinc citrate, but is not limited thereto.

[0057] In the present invention, "cyclo-hisPro (CHP)" refers to a naturally occurring circular dipeptide composed of histidine-proline, a metabolite of thyrotropin-releasing hormone (TRH), or a physiologically active dipeptide synthesized de novo in the body through the TRH metabolic process, and is a substance widely distributed throughout the brain, spinal cord, and gastrointestinal tract.

[0058] In the present invention, the cyclo-hispro may be synthesized or commercially available. Furthermore, it may be purified from substances containing cyclo-hispro, such as prostate extracts and soybean hydrolysates.

[0059] The term "purified" is intended to indicate that cyclo-hisprolate is in a more concentrated form than its natural source, such as prostate extract. Purified ingredients can be concentrated from their natural sources or obtained through chemical synthesis.

[0060] In the present invention, the zinc salt and cyclo-hispro are also referred to as "Cyclo-Z", and the zinc salt and cyclo-hispro can be included in the composition of the present invention in the form of a single complex or as individual components. At this time, the weight ratio of the zinc salt: cyclo-hispro can be 1 to 10: 1 to 5, and preferably 1 to 5: 1 to 2, but is not limited thereto. In the present invention, the weight ratio for the zinc salt represents the amount of zinc cations or the weight ratio of the zinc component in the zinc salt.

[0061] Therefore, zinc salt and cyclo-hispro can be administered in the form of a single complex or as individual components, administered simultaneously, separately or sequentially.

[0062] In the present invention, the pharmaceutical composition may additionally contain an antihypertensive agent as another effective ingredient.

[0063] Additionally, in the present invention, the composition administered to the subject may be administered in combination with an antihypertensive agent.

[0064] In the present invention, the antihypertensive agent may be an ARB or an ACE inhibitor. The ARB and ACE are antihypertensive agents that inhibit the action of the renin-angiotensin-aldosterone system (RAAS). ARB blocks the action of angiotensin II on angiotensin II receptor type 1 (AT1), and ACE inhibitors inhibit ACE to block the conversion of angiotensin I to angiotensin II.

[0065] In the present 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.

[0066] In the present invention, the ACE inhibitor may be any one selected from the group consisting of Captopril, Zofenopril, Enalapril, Ramipril, Quinapril, Perindopril, Lisinopril, Benazepril, Imidapril, Trandolapril, Cilazapril, and Fosinopril, but is not limited thereto.

[0067] In the present invention, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z): antihypertensive agent may be 1 to 2: 1 to 4, and the antihypertensive agent may be at a concentration equal to or higher than that of the zinc salt and cyclo-hispro (Cyclo-Z), but is not limited thereto.

[0068] Angiotensin II, a major end product of the renin-angiotensin system (RAS), is known to play a central role in the development and progression of cardiovascular diseases at various points in the cardiovascular disease continuum by binding to the AT1 receptor and causing vasoconstriction, increased sodium absorption in the renal tubules, water retention, impaired endothelial function, increased oxidative stress, and increased tissue fibrosis and collagen deposition. Angiotensin II receptor blockers (ARBs) are currently the most widely used drugs for the treatment of hypertension due to their excellent hypotensive effects and minimal side effects through physiological blockade of the AT1 receptor of the RAS. However, despite these various benefits, concerns have been raised about their safety, such as the so-called ARB-myocardial infarction paradox, which suggests that the use of ARBs may actually increase the risk of myocardial infarction, and the controversy over an increased incidence of cancer.

[0069] In the present invention, the pharmaceutical composition may additionally contain an antidiabetic agent as another effective ingredient.

[0070] Additionally, in the present invention, the composition administered to the subject may be administered in combination with an antidiabetic agent.

[0071] In the present invention, the antidiabetic agent may be any one selected from the group consisting of an SGLT2 inhibitor, a DPP-4 inhibitor, and a biguanide drug.

[0072] In the present invention, the SGLT2 inhibitor may be any one selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin, but is not limited thereto.

[0073] In the present invention, the DPP-4 inhibitor may be any one selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, anagliptin, and evogliptin, but is not limited thereto.

[0074] The above biguanide drug may be any one selected from the group consisting of metformin, buformin, and phenformin, but is not limited thereto.

[0075] In the present invention, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z): SGLT2 inhibitor may be 5 to 10: 1 to 1.5, but is not limited thereto.

[0076] In the present invention, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z): DPP-4 inhibitor may be 1 to 3.5: 1 to 2, wherein the DPP-4 inhibitor may be at the same or lower concentration as the zinc salt and cyclo-hispro (Cyclo-Z), but is not limited thereto.

[0077] In the present invention, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z):biguanide drug may be 1 to 2:10 to 16, but is not limited thereto.

[0078] In the present invention, the pharmaceutical composition may additionally include both the antihypertensive agent and the antidiabetic agent as other effective ingredients. In this case, since the description of the antihypertensive agent and the antidiabetic agent is the same as described above, the description thereof is omitted.

[0079] In addition, in the present invention, the composition administered to the subject may be administered in combination with an antihypertensive agent and an antidiabetic agent. In this case, the description of the antihypertensive agent and the antidiabetic agent is the same as described above, and thus, description thereof is omitted.

[0080] In the present invention, when the pharmaceutical composition comprises a zinc salt and cyclo-hispro (Cyclo-Z), an antihypertensive agent, and an SGLT2 inhibitor, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z): antihypertensive agent: SGLT2 inhibitor may be 5 to 10: 5 to 10: 0.5 to 1, and wherein the antihypertensive agent may be at the same concentration as or higher than that of the zinc salt and cyclo-hispro (Cyclo-Z), but is not limited thereto.

[0081] In the present invention, when the pharmaceutical composition comprises a zinc salt and cyclo-hispro (Cyclo-Z), an antihypertensive agent, and a DPP-4 inhibitor, the weight ratio of the zinc salt and cyclo-hispro (Cyclo-Z): antihypertensive agent: DPP-4 inhibitor may be 5 to 10: 5 to 10: 2 to 5, wherein the antihypertensive agent may be at a concentration equal to or higher than that of the zinc salt and cyclo-hispro (Cyclo-Z), and the DPP-4 inhibitor may be at a concentration lower than that of the zinc salt and cyclo-hispro (Cyclo-Z) and the antihypertensive agent, but is not limited thereto.

[0082] In the present invention, zinc salt and cyclo-hispro (Cyclo-Z) may be included in a single complex form at a dosage of 15 to 100 mg / kg when applied clinically, for example, 38 mg / kg, 76 mg / kg, or 91 mg / kg per dosage unit, but is not limited thereto. If zinc salt and cyclo-hispro are included as individual components, they may be included at a dosage calculated according to the weight ratio of zinc salt: cyclo-hispro defined in the present invention within the above dosage range.

[0083] In the present invention, the SGLT2 inhibitor may be included in a dosage of 2 to 15 mg / kg when applied clinically, for example, it may be included in a dosage of 5 mg / kg or 10 mg / kg per dosage unit, but is not limited thereto.

[0084] In the present invention, the DPP-4 inhibitor may be included in a dosage of 10 to 120 mg / kg for clinical application. For example, it may be included in a dosage of 25 mg / kg, 50 mg / kg, or 100 mg / kg per dosage unit, but is not limited thereto.

[0085] In the present invention, the biguanide drug may be included in a dosage of 200 to 1500 mg / kg for clinical use. For example, it may be included in a dosage of 500 mg / kg or 1000 mg / kg per dosage unit, but is not limited thereto.

[0086] As side effects of existing antihypertensive and antidiabetic agents have been reported, the present inventors sought to search for a substance that can prevent or reduce side effects caused by overdose or long-term administration of antihypertensive and / or antidiabetic agents by replacing the combined administration of these drugs or by significantly enhancing the therapeutic effect of these drugs in diabetic nephropathy. As a result, as confirmed in Figures 1a to 1d, the combination of zinc salt and cyclo-hispro not only showed an albuminuria-reducing effect in the KKay diabetic nephropathy mouse model, but also significantly enhanced the albuminuria-reducing effect of these drugs when co-administered with losartan, a representative ARB, and / or dapagliflozin, a representative SGLT2 drug.

[0087] In addition, as confirmed in Figures 2a to 2c, it was confirmed that the combination of zinc salt and cyclo-hispro (CycloZ) not only showed an ACR reducing effect in the KKay diabetic nephropathy mouse model, but also significantly increased the ACR reducing effect of these drugs when co-administered with losartan and / or dapagliflozin.

[0088] Additionally, as confirmed in Table 2, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclo-hispro was co-administered with losartan and / or dapagliflozin, and it was confirmed that the glomerular filtration rate-increasing effect of these drugs was further enhanced.

[0089] In addition, as confirmed in Figures 3a and 3b, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclo-hispro, when co-administered with sitagliptin, a representative drug of DPP-4 inhibitors, significantly increased the ACR reduction effect compared to sitagliptin monoadministration, regardless of the severity of diabetic nephropathy.

[0090] Furthermore, as confirmed in Fig. 4, in the KKay diabetic nephropathy mouse model, the combination of cyclo-hispro significantly increased the ACR reduction effect when co-administered with losartan, a representative ARB drug, and sitagliptin, a representative DPP-4 inhibitor drug, compared to co-administration of losartan and sitagliptin.

[0091] In addition, as confirmed in Fig. 5, in the KKay diabetic nephropathy mouse model, the combination of cyclo-hispro and metformin, a representative drug of biguanide, significantly increased the ACR reduction effect compared to administration of cyclo-hispro alone and metformin alone.

[0092] Additionally, as confirmed in Fig. 6, in the KKay diabetic nephropathy mouse model, the combination of zinc salt and cyclo-hispro (CycloZ) significantly reduced the ACR level compared to cyclo-hispro alone, confirming the synergistic effect of co-administration of zinc salt and cyclo-hispro.

[0093] Accordingly, the pharmaceutical composition of the present invention can exhibit one or more effects selected from the group consisting of i) reducing albuminuria; ii) reducing albuminuria / creatinine ratio; and iii) increasing glomerular filtration rate.

[0094] In the present invention, the diabetic nephropathy includes both type 1 diabetic nephropathy and type 2 diabetic nephropathy, and the effects of i) to iii) can be equally exhibited regardless of the type of diabetes.

[0095] The combination of zinc salt and cyclo-hyspro is known to have a therapeutic effect by reducing blood sugar in type 2 diabetes, which is non-insulin dependent diabetes, but does not show a distinct therapeutic effect such as reducing blood sugar in type 1 diabetes, which is insulin dependent diabetes. However, in diabetic nephropathy, it shows the effects of reducing albuminuria, reducing the albuminuria / creatinine ratio, and increasing the glomerular filtration rate regardless of the type of diabetes (i.e., both type 1 and type 2 diabetes), and thus shows the effect of preventing, improving, or treating diabetic nephropathy through an independent activity separate from the effect of treating type 2 diabetes.

[0096] In the present invention, the pharmaceutical composition may be administered simultaneously, separately or sequentially with the antihypertensive agent and / or the antidiabetic agent.

[0097] In the present invention, the term "synergistic effect" refers to an effect that occurs when each component is administered in combination (in combination) and is greater than the sum of the effects that occur when each component is administered alone. The composition comprising the zinc salt of the present invention and cyclo-hispro or a pharmaceutically acceptable salt thereof enhances the preventive, ameliorating, or therapeutic effect on diabetic nephropathy when administered in combination with an antihypertensive agent and / or an antidiabetic agent.

[0098] The term "administered in combination" refers to compounds or components being administered to a patient together. Coadministration of each compound or component means that each component may be administered at the same time, in any order, or sequentially at different times to achieve the desired therapeutic effect.

[0099] The term "prevention" as used herein refers to any action that inhibits or delays the onset of a disease or condition. In the present invention, it refers to delaying or inhibiting the onset of diabetic nephropathy.

[0100] The term "improvement" used in the present invention means any act of improving or beneficially changing a disease or condition, and in the present invention means improving the symptoms of diabetic nephropathy.

[0101] The term "treatment" used in the present invention means any act of delaying, stopping, or reversing the progression of a disease or condition, and in the present invention means alleviating, alleviating, eliminating, or reversing the symptoms of diabetic nephropathy.

[0102] The term "pharmaceutically acceptable salt" used in the present invention means any organic or inorganic addition salt of cyclo-hispro, which has an effective effect that is relatively non-toxic and harmless to the patient, and the side effects caused by the salt do not reduce the beneficial efficacy of cyclo-hispro. These salts may use inorganic acids and organic acids as free acids, and inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc. may be used, and organic acids such as citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid, or malonic acid may be used. Additionally, these salts include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, acid addition salts include acetate, aspartate, benzate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, saccharate, stearate, succinate, May contain tartrate, tosylate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, and zinc salts.

[0103] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The pharmaceutical composition comprising the pharmaceutically acceptable carrier may be in various oral or parenteral dosage forms. When formulated, it may be prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration may include tablets, tablets, powders, granules, capsules, troches, etc., and such solid preparations may be prepared by mixing one or more compounds of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration may include suspensions, solutions, emulsions, or syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives may be included.

[0104] Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.

[0105] The pharmaceutical composition of the present invention may be administered via any conventional route capable of reaching the target tissue or cell within a subject or sample. Such 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, dosage form, health status, and disease severity.

[0106] In the methods of the present invention, the term "subject" includes, but is not limited to, any animal (e.g., a 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 gender. Thus, it is intended to include female / female or male / male, adult / adult and newborn subjects, as well as fetuses. A patient refers to a subject suffering from a disease or disorder. The term patient includes human and veterinary subjects.

[0107] In the method of the present invention, the description of the composition including the effects, administration route, number of administrations, dosage, etc. of the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof administered to the subject is the same as described above, and therefore, description thereof is omitted.

[0108] In the method of the present invention, when administered in an effective amount, a zinc salt and cyclo-hispro can provide a desirable effect of preventing, improving, or treating diabetic nephropathy. For a desirable effect, the combination of the zinc salt of the present invention and cyclo-hispro or a pharmaceutically acceptable salt thereof may be administered once or repeatedly several times at regular time intervals. In this case, the zinc salt and cyclo-hispro may be administered simultaneously, separately, or sequentially. For example, the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof may be co-formulated and administered simultaneously as a combined unit dosage form, or may be administered simultaneously or sequentially as separate formulations.

[0109] Alternatively, a composition comprising a zinc salt and cyclo-hispro may provide a desirable effect of preventing, improving, or treating diabetic nephropathy when administered in combination with an antihypertensive and / or antidiabetic agent in an effective amount. For a desirable effect, the combination of the composition comprising the zinc salt of the present invention and cyclo-hispro or a pharmaceutically acceptable salt thereof and the antihypertensive and / or antidiabetic agent may be administered once or repeatedly several times at regular time intervals. In this case, the combination of the composition comprising the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof and the antihypertensive and / or antidiabetic agent may be administered simultaneously, separately, or sequentially. For example, a combination of a composition comprising the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof and an antihypertensive and / or antidiabetic agent may be co-formulated and administered simultaneously as a combined unit dosage form, or may be administered simultaneously or sequentially as separate formulations.

[0110] When administered sequentially, each active ingredient may be administered as an individual formulation at intervals of time, and the order of administration may be determined by a physician or a person having ordinary knowledge in the field.

[0111] Additionally, it can be used in combination with other methods to prevent, improve or treat diabetic nephropathy.

[0112] In the use of the present invention, the effect of the composition containing the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof and the description of the composition including the route of administration, number of administrations, dosage, etc. are the same as those described above, and therefore, description thereof is omitted.

[0113] The composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients according to the present invention significantly increases the therapeutic effect of diabetic nephropathy when administered in combination with an ARB and / or an SGLT2 inhibitor, and can also be utilized as an adjuvant for improving the therapeutic effect of these drugs on diabetic nephropathy.

[0114] Accordingly, the second aspect of the present invention relates to a pharmaceutical composition for enhancing the preventive, ameliorating or therapeutic effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy, the pharmaceutical composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients.

[0115] According to one embodiment of the present invention, a pharmaceutical composition for improving the therapeutic effect of an ARB and / or an SGLT2 inhibitor on diabetic nephropathy is provided, the pharmaceutical composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof as active ingredients.

[0116] The composition and effect of zinc salt and cyclo-hispro, which are included as active ingredients in the pharmaceutical composition for improving the diabetic nephropathy treatment effect of the ARB and / or SGLT2 inhibitor according to the present invention, are the same as described above, and therefore, description thereof is omitted.

[0117] In addition, the types of antihypertensive and antidiabetic agents (specifically, ARB and SGLT2) that can exhibit a synergistic effect when combined with the zinc salt and cyclo-hispro are the same as those described above, and therefore, their description is omitted.

[0118] In relation to the second aspect, the present invention provides a method for improving the effect of preventing, improving, or treating diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof. In this case, the subject is the same as described in the first aspect, and thus description thereof is omitted. Alternatively, the subject may be a patient who exhibits side effects due to overdose or long-term administration of an antihypertensive and / or antidiabetic agent, or is expected to exhibit such side effects. For example, the subject may be a diabetic nephropathy patient who is receiving an antihypertensive and / or antidiabetic agent, preferably an ARB and / or an SGLT2 inhibitor.

[0119] In relation to the second aspect, the present invention also provides a use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof, for use in preventing, improving or enhancing the effect of treating diabetic nephropathy.

[0120] In relation to the second aspect, the present invention also provides the use of a composition comprising a zinc salt; and cyclo-hispro or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing, improving or enhancing the effect of treating diabetic nephropathy.

[0121] In the present invention, the composition comprising the zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof may be administered in combination with an antihypertensive and / or antidiabetic agent for the purpose of enhancing the preventive, ameliorating, or therapeutic effect of the antihypertensive and / or antidiabetic agent on diabetic nephropathy. In this case, the description of the antihypertensive and antidiabetic agents is the same as that described in the first aspect, and thus, description thereof is omitted.

[0122] The third aspect of the present invention relates to a health functional food composition for preventing or improving diabetic nephropathy, comprising zinc salt and cyclo-hispro or a food-related acceptable salt thereof as active ingredients.

[0123] In relation to the third aspect, the present invention also provides a use of a composition comprising a zinc salt and cyclo-hispro or a food-wise acceptable salt thereof for the manufacture of a health functional food for preventing, improving or enhancing the effect of treating diabetic nephropathy.

[0124] In the present invention, the composition comprising the zinc salt and cyclo-hispro or a food-based acceptable salt thereof may be administered in combination to a diabetic nephropathy patient receiving an antihypertensive and / or antidiabetic agent for the purpose of enhancing the preventive, ameliorating, or therapeutic effect of the antihypertensive and / or antidiabetic agent in diabetic nephropathy. In this case, the description of the antihypertensive and antidiabetic agents is the same as that described in the first aspect, and thus, description thereof is omitted.

[0125] According to one embodiment of the present invention, the health functional food composition may be for improving the effect of improving diabetic nephropathy in a diabetic nephropathy patient receiving an antihypertensive and / or antidiabetic agent, for example, an ARB and / or an SGLT2 inhibitor.

[0126] In the health functional food composition of the present invention, the description of the composition and effect of zinc salt and cyclo-hispro contained as active ingredients is the same as described above, and therefore, description thereof is omitted.

[0127] In the present invention, the term "food-acceptable salt" includes salts derived from food-acceptable organic acids, inorganic acids, or bases. The types of food-acceptable salts are identical to the types of "pharmaceutically acceptable salts" described in the first aspect, and therefore, their description is omitted.

[0128] In the present invention, the term “health functional food” includes both the meanings of “functional food” and “health food.”

[0129] In the present invention, the term "functional food" is the same as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition.

[0130] In the present invention, the term "health food" refers to a food that has a more active health maintenance or promotion effect than regular food, and "health supplement food" refers to a food for health supplement purposes. In some cases, the terms "functional food," "health food," and "health supplement food" may be used interchangeably. The above food may be manufactured in various forms, such as tablets, capsules, powders, granules, liquids, and pills, to obtain beneficial effects in preventing or improving diabetic nephropathy.

[0131] As a specific example of such functional foods, by using the zinc salt and cyclo-hispro of the present invention or a food-wise acceptable salt thereof, a processed food can be manufactured that improves the storage properties of agricultural products, livestock products or marine products while modifying them to preserve their characteristics.

[0132] The health functional food composition of the present invention can also be manufactured in the form of a nutritional supplement, food additive, feed, etc., and is intended for consumption by humans or animals including livestock.

[0133] The above type of food composition can be manufactured in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats and processed foods thereof (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juices, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible plant oils, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.), etc., which can be manufactured by adding the zinc salt and cyclo-hispro of the present invention or a food-wise acceptable salt thereof.

[0134] In addition, nutritional supplements may be prepared by adding the zinc salt and cyclo-hispro or a food-wise acceptable salt thereof of the present invention to capsules, tablets, pills, etc., but are not limited thereto.

[0135] In addition, the health functional food is not limited thereto, but for example, the zinc salt and cyclo-hispro or a food-scientifically acceptable salt thereof of the present invention can be manufactured in the form of tea, juice, and drink, and can be consumed by liquefying, granulating, encapsulating, and powdering so that they can be consumed (health drinks). In addition, in order to use the zinc salt and cyclo-hispro or a food-scientifically acceptable salt thereof of the present invention in the form of a food additive, they can be manufactured in the form of a powder or concentrate. In addition, the zinc salt and cyclo-hispro or a food-scientifically acceptable salt thereof of the present invention can be manufactured in the form of a composition by mixing with a known active ingredient known to be effective in preventing or improving diabetic nephropathy.

[0136] When the food composition of the present invention is used as a health beverage composition, the health beverage composition may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. The natural carbohydrates described above may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin and stevia extract; or a synthetic sweetener such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

[0137] The zinc salt and cyclo-hispro or a food-wise acceptable salt thereof of the present invention may be contained as an active ingredient of a health functional food composition for preventing or improving diabetic nephropathy, and the amount thereof is an amount effective to obtain the above-mentioned preventive or improving effect, for example, preferably 0.01 to 100 wt% based on the total weight of the entire composition, but is not particularly limited thereto. The health functional food composition of the present invention may be prepared in the form of a composition by mixing a known active ingredient known to be effective in preventing or improving diabetic nephropathy together with the zinc salt and cyclo-hispro or a food-wise acceptable salt thereof.

[0138] In addition to the above, the health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, or carbonating agents. In addition, the health functional food of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in mixtures. The proportion of these additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0139] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0140] [Example 1]

[0141] Confirmation of the effect of CycloZ, ARB / ACE inhibitor and / or SGLT2 inhibitor administration on reducing albuminuria and albuminuria / creatinine ratio in diabetic animal models.

[0142] 1-1. Experimental animals and administered drugs

[0143] Five-week-old KKay mice were purchased from Clea, Japan, and 5-week-old C57BL6 / J mice were purchased from Daehan Biolink as normal controls. Rearing was performed under constant conditions (temperature: 22±2℃, relative humidity: 55±10%, circadian rhythm: 12 h), and Purina mouse chow was used as feed and distilled water as drinking water. Mice were used in the experiment after a one-week adaptation period after purchase. Losartan used in the examples below was purchased from Angene, and dapagliflozin was purchased from Betapahrma. Metabolic cages were prepared from Jeongdo B&P Co., Ltd. Isoflurane, required for mouse anesthesia and dissection, was purchased from Hana Pharmaceutical, and a Vetequip RC2 Rodent Circuit Controller Anesthesia System was prepared. Phosphate buffered saline (PBS) was purchased from Hyclone.

[0144] The KKAy animal model is known to have 10-50 times more proteinuria than the db / db animal model, and mesangial matrix expansion has been reported to be more severe than all models. Five-week-old KKAy mice purchased from Clea Japan were pre-fed for 1 week and then divided into groups as shown in Table 1. Losartan was dissolved in drinking water and administered per animal at a daily dose of 20 mg / kg, and dapagliflozin (2 mg / kg), CycloZ (15 mg / kg), or their combination drugs were administered orally daily. The normal control group and the negative control group were administered orally the same amount of distilled water daily.

[0145] Group Administration DrugNormal Control Distilled WaterNegative Control Distilled WaterGroup 1 Losartan 20 mg / kgGroup 2 Dapagliflozin 2 mg / kgGroup 3 CycloZ 15 mg / kg (CHP 5 mg / kg + Zn 10 mg / kg)Group 4 Losartan 20 mg / kg + CycloZ 15 mg / kg (CHP 5 mg / kg + Zn 10 mg / kg)Group 5 Dapagliflozin 2 mg / kg + CycloZ 15 mg / kg (CHP 5 mg / kg + Zn 10 mg / kg)Group 6 Losartan 20 mg / kg + Dapagliflozin 2 mg / kgGroup 7 Losartan 20 mg / kg + Dapagliflozin 2 mg / kg + CycloZ 15 mg / kg (CHP 5 mg / kg + Zn 10 mg / kg)

[0146] 1-2. Measurement of albuminuria

[0147] At week 16 of administration, 24-hour urine samples were collected using metabolic cages and centrifuged at 2,000 rpm for 10 minutes to remove suspended solids. Albumin was measured using a DCA vantage analyzer (Siemens) and DCA® Microalbumin / Creatinine Urine Test cartridge (Siemens) using urine diluted 1:10 with distilled water.

[0148] As a result, as shown in Fig. 1a, albuminuria (mg / dL) was detected as 51.4 in the negative control group, 39.3 in the Losartan monotherapy group (Group 1), 45.4 in the CycloZ monotherapy group (Group 3), and 11.1 in the Losartan + CycloZ (Losartan + CycloZ) combination administration group (Group 4), and each drug administration group showed a 23.5%, 11.7%, and 78.4% reduction in albuminuria compared to the negative control group. From the above results, it was confirmed that the Losartan + CycloZ combination administration showed a synergistic effect in reducing albuminuria.

[0149] In addition, as shown in Fig. 1b, albuminuria (mg / dL) was detected as 51.4 in the negative control group, 31.1 in the dapagliflozin monotherapy group (group 2), 45.4 in the CycloZ monotherapy group (group 3), and 22.4 in the dapagliflozin + CycloZ combination therapy group (group 5), and each drug administration group showed a decrease of 39.5%, 11.7%, and 56.4% compared to the negative control group. From the above results, it was confirmed that dapagliflozin + CycloZ combination therapy showed a synergistic effect in reducing albuminuria.

[0150] In addition, as shown in Fig. 1c, albuminuria (mg / dL) was detected as 51.4 in the negative control group, 45.4 in the CycloZ monotherapy group (Group 3), 6.7 in the Losartan+Dapagliflozin combination therapy group (Group 6), and 2.2 in the Lorsaltan+Dapagliflozin+CycloZ combination therapy group (Group 7), and each drug administration group showed a reduction of 11.7%, 87%, and 95.7% compared to the negative control group. From the above results, it was confirmed that the Losartan+Dapagliflozin+CycloZ combination therapy showed a synergistic effect in reducing albuminuria.

[0151] Lastly, Fig. 1d shows the results of comparing all albuminuria measurements of groups 1 to 7, and albuminuria (mg / dL) was detected as 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 CycloZ monotherapy group (group 3), 11.1 in the losartan + CycloZ combination group (group 4), 22.4 in the dapagliflozin + CycloZ combination group (group 5), 6.7 in the losartan + dapagliflozin combination group (group 6), and 2.2 in the losartan + dapagliflozin + CycloZ combination group (group 7), respectively, compared to the negative control group at 23.5%, 39.5%, 11.7%, and 56.4%, respectively. A reduction of 78.4%, 87%, and 95.7% was observed. In particular, the combination of losartan + dapagliflozin + CycloZ showed a significant albuminuria reduction effect compared to the individual or combination administration groups of the two drugs, confirming that the synergistic effect of the three combination administrations was the most excellent.

[0152] 1-3. Check the albuminuria / creatinine ratio

[0153] Creatinine was measured using undiluted urine using a mouse creatinine assay kit (Crystalchem). The measured albumin (mg / dL) value was divided by the creatinine (g / dL) value to derive the ACR (mg / g).

[0154] As a result, as shown in Fig. 2a, it was confirmed that the ACR value of the Losartan + CycloZ combination administration group (Group 4) decreased more than that of the groups administered Losartan or CycloZ alone (Group 1 or Group 3), and that a synergistic effect was observed due to combination administration.

[0155] In addition, as shown in Figure 2b, it was confirmed that the ACR value of the dapagliflozin + CycloZ combination administration group (group 5) was further reduced compared to the groups administered dapagliflozin or CycloZ alone (group 2 or group 3), and a synergistic effect was observed due to the combination administration.

[0156] Finally, as shown in Figure 2c, it was confirmed that the ACR value of the Losartan + Dapagliflozin + CycloZ combination administration group (Group 7) was significantly reduced compared to the groups administered with Losartan, Dapagliflozin, or CycloZ alone (Groups 1 to 3), and was further reduced compared to the Losartan + Dapagliflozin combination administration group.

[0157] [Example 2]

[0158] Confirmation of the effect of increasing glomerular filtration rate in diabetic animal models

[0159] At week 17 of administration, the hair on the back of the mice was shaved with a hair caliper, Nair hair removal cream was applied, and after 5 minutes, the hair was removed by wiping it with a gauze soaked in warm water. The following day, a GFR monitoring device (Medibeacon) was attached to the back of the mice, and FITC-sinistrin (Medibeacon) dissolved in physiological saline was injected into the tail vein at a concentration of 150 mg / kg. After 90 minutes of real-time measurement, the device was removed, and GFR values ​​were analyzed using Medibeacon Studio software.

[0160] As a result, as shown in Table 2, the GFR value of the Losartan + Dapagliflozin + CycloZ combination administration group (Group 7) was significantly increased compared to the negative control group, and was further increased compared to the Losartan + CycloZ combination administration group (Group 4) and the Dapagliflozin + CycloZ combination administration group (Group 5).

[0161] Group GFR Negative control group 0.580 Losartan + CycloZ 0.839 Dapagliflozin + CycloZ 0.826 Losartan + Dapagliflozin + CycloZ 0.932

[0162] [Example 3]

[0163] Confirmation of the effect of reducing the albuminuria / creatinine ratio following administration of CycloZ and / or DPP-4 inhibitors in diabetic animal models

[0164] 3-1. Administration of medication for mild conditions

[0165] Five-week-old KKAy mice from Example 1-1 were bred for one week and then randomly divided into groups as shown in Table 3 so that the average body weights would be the same. The control group was orally administered distilled water for 11 weeks, and Group 1 was orally administered CycloZ 15 mg / kg, Group 2 was orally administered sitagliptin 10 mg / kg, and Group 3 was orally administered sitagliptin 10 mg / kg + CycloZ 15 mg / kg daily for the same period.

[0166] Group Dosage Drug Negative Control Group (CTRL) Distilled water Group 1 CycloZ 15 mg / kg Group 2 Sitagliptin 10 mg / kg Group 3 Sitagliptin 10 mg / kg + CycloZ 15 mg / kg

[0167] 3-2. Confirmation of albuminuria / creatinine ratio in mild cases

[0168] At week 11 of 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 a DCA vantage analyzer (Siemens) and DCA® Microalbumin / Creatinine Urine Test cartridges (Siemens) using urine diluted 1 / 2 with distilled water. Creatinine was measured using a mouse creatinine assay kit (Crystalchem) using undiluted urine. The measured albumin (mg / dL) value was divided by the creatinine (g / dL) value to derive the ACR (mg / g).

[0169] As a result, as confirmed in Fig. 3a, 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 group. These values ​​were reduced by approximately 37%, 24%, and 63%, respectively, compared to the control group. As the ACR value of the sitagliptin + CycloZ combination group showed a significantly improved ACR reduction effect compared to other monotherapy groups, it was confirmed that it showed a synergistic effect by CycloZ.

[0170] 3-3. Administration of medication for severe conditions

[0171] Five-week-old KKAy mice from Example 1-1 were bred for six weeks, and urine was collected for 24 hours, and ACR was measured using the same method as in Example 3-2. The mice were randomly divided into groups as shown in Table 4 so that the average ACR values ​​were the same, and then, starting at 12 weeks of age, drugs were administered orally daily for 11 weeks as shown in Table 4.

[0172] Group Dosage Drug Negative Control Group (CTRL) Distilled water Group 1 CycloZ 15 mg / kg Group 2 Sitagliptin 10 mg / kg Group 3 Sitagliptin 10 mg / kg + CycloZ 15 mg / kg

[0173] 3-4. Check the albuminuria / creatinine ratio in severe cases.

[0174] At the 11th week of administration, urine was collected for 24 hours using a metabolic cage, and ACR was measured using the same method as in Example 3-2.

[0175] As a result, as confirmed in Fig. 3b, 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 group. In administration from the severe stage, CycloZ monotherapy did not show a significant effect on improving ACR, but the sitagliptin monotherapy group showed a 48% decrease compared to the negative control group, and the sitagliptin + CycloZ combination group showed a 31% decrease compared to the sitagliptin monotherapy group and a 64% decrease compared to the negative control group. Through this, it was possible to confirm the synergistic effect of sitagliptin + CycloZ combination administration in improving severe stage diabetic nephropathy.

[0176] [Example 4]

[0177] Confirmation of the effect of CycloZ, ARB / ACE inhibitor, and / or DPP-4 inhibitor administration on reducing the albuminuria / creatinine ratio in diabetic animal models.

[0178] 4-1. Administration of medication for severe conditions

[0179] Five-week-old KKAy mice from Example 1-1 were raised for six weeks, and urine was collected for 24 hours, and ACR was measured using the same method as in Example 3-2. The mice were randomly divided into groups as shown in Table 5 so that the average ACR values ​​were the same, and drugs were administered for nine weeks starting at 12 weeks of age as shown in Table 5. At this time, losartan was dissolved in drinking water and ingested per animal daily at 20 mg / kg, and the remaining drugs were administered orally daily.

[0180] Group Administration Drug Negative Control Group (CTRL) Distilled water Group 1 CycloZ 15 mg / kg Group 2 Losartan 20 mg / kg Group 3 Losartan 20 mg / kg + CycloZ 15 mg / kg Group 4 Losartan 20 mg / kg + Sitagliptin 10 mg / kg Group 5 Losartan 20 mg / kg + Sitagliptin 10 mg / kg + CycloZ 15 mg / kg

[0181] 4-2. Confirmation of albuminuria / creatinine ratio in severe conditions

[0182] At the 9th week of administration, urine was collected for 24 hours using a metabolic cage, and ACR was measured using the same method as in Example 3-2.

[0183] As a result, as confirmed in Fig. 4, 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 group. In the severe stage, CycloZ monotherapy and Losartan monotherapy did not show a significant effect on improving ACR, but the Losartan + CycloZ combination group showed a 42% decrease compared to the negative control group. This confirmed the synergistic effect of Losartan + CycloZ combination administration in improving severe stage diabetic nephropathy.

[0184] In addition, as confirmed in Fig. 4, the ACR value of the Losartan + Sitagliptin combination administration group was 2,768 mg / g, and the Losartan + Sitagliptin + CycloZ combination administration group was 2,025 mg / g, and these values ​​were reduced by 61.1% and 71.6%, respectively, compared to the negative control group. As the ACR value of the Losartan + Sitagliptin + CycloZ combination administration group was reduced more than that of the Losartan + Sitagliptin combination administration group, it was confirmed that there was a synergistic effect due to CycloZ.

[0185] [Example 5]

[0186] Confirmation of the effect of CycloZ and / or biguanide drug administration on reducing the albuminuria / creatinine ratio in diabetic animal models.

[0187] 5-1. Administration of medication for severe conditions

[0188] Five-week-old KKAy mice from Example 1-1 were raised for six weeks, and urine was collected for 24 hours, and ACR was measured using the same method as in Example 3-2. The mice were randomly divided into groups as shown in Table 6 so that the average ACR values ​​were the same, and then, starting at 12 weeks of age, drugs were administered orally daily for 12 weeks as shown in Table 6.

[0189] Group administration drug negative control group (CTRL) distilled water group 1 CycloZ 15 mg / kg group 2 metformin 200 mg / kg group 3 metformin 200 mg / kg + CycloZ 15 mg / kg

[0190] 5-2. Confirmation of albuminuria / creatinine ratio in severe conditions

[0191] At the 12th week of administration, urine was collected for 24 hours using a metabolic cage, and ACR was measured using the same method as in Example 3-2.

[0192] As a result, as confirmed in Fig. 5, the ACR (mg / g) value was 2,369 mg / g in the negative control group, 2,792 mg / g in the CycloZ alone administration group, 2,952 mg / g in the metformin alone administration group, and 1,117 mg / g in the metformin + CycloZ combination administration group. In administration from the severe stage, CycloZ alone and metformin alone did not show a significant effect on improving ACR, but the metformin + CycloZ combination administration group showed a 53% decrease compared to the negative control group. Through this, it was possible to confirm the synergistic effect of metformin + CycloZ combination administration in improving severe stage diabetic nephropathy.

[0193] [Example 6]

[0194] Confirmation of the effect of reducing the albuminuria / creatinine ratio by a combination of zinc salt and CHP in a diabetic animal model.

[0195] The same animal models used in Example 1 were pre-fed for one week and then divided into groups as shown in Table 7. CHP (5 mg / kg or 35 mg / kg), CycloZ (15 mg / kg), and bardoxolone methyl (mg / kg) were administered orally daily, respectively. The normal control group and the negative control group were administered the same amount of distilled water orally daily.

[0196] Group administration drug Normal control group Distilled water Negative control group Distilled water Group 1 CHP 5 mg / kg Group 2 CHP 35 mg / kg Group 3 CycloZ 15 mg / kg (CHP 5 mg / kg + Zn 10 mg / kg) Positive control group Bardoxolone methyl (10 mg / kg)

[0197] Creatinine was measured using a mouse creatinine assay kit (Crystalchem) using undiluted urine. The measured albumin (mg / dL) value was divided by the creatinine (g / dL) value to derive the ACR (mg / g) value. As a result, as shown in Figure 6, the ACR value of the CycloZ administration group (group 3) was significantly reduced compared to the groups administered CHP alone (group 1 or group 2), confirming a synergistic effect of co-administration of CHP and Zn compared to CHP administration alone.

[0198] While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Zinc salt; and A pharmaceutical composition for preventing or treating diabetic nephropathy, comprising cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

2. A pharmaceutical composition for preventing or treating diabetic nephropathy, comprising an antihypertensive agent selected from the group consisting of an angiotensin II receptor blocker and an angiotensin-converting enzyme inhibitor as an additional active ingredient in paragraph 1.

3. A pharmaceutical composition for preventing or treating diabetic nephropathy, comprising an antidiabetic agent selected from the group consisting of a sodium glucose cotransporter 2 inhibitor, a dipeptidyl peptidase-4 inhibitor, and a biguanide drug as an additional active ingredient in paragraph 1.

4. A pharmaceutical composition for preventing or treating diabetic nephropathy, comprising as an additional active ingredient an antihypertensive agent selected from the group consisting of an angiotensin II receptor blocker and an angiotensin-converting enzyme inhibitor; and an antidiabetic agent selected from the group consisting of a sodium glucose cotransporter 2 inhibitor, a dipeptidyl peptidase-4 inhibitor, and a biguanide drug.

5. In paragraph 2 or 4, the angiotensin II receptor blocker is any one selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, fimasartan, irbesartan, eprosartan, and azilsartan, A pharmaceutical composition for preventing or treating diabetic nephropathy, wherein the angiotensin converting enzyme inhibitor is any one selected from the group consisting of Captopril, Zofenopril, Enalapril, Ramipril, Quinapril, Perindopril, Lisinopril, Benazepril, Imidapril, Trandolapril, Cilazapril, and Fosinopril.

6. In the third or fourth paragraph, the sodium glucose cotransporter-2 inhibitor is any one selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin, The above dipeptidyl peptidase-4 inhibitor is any one selected from the group consisting of Sitagliptin, Linagliptin, Vildagliptin, Gemigliptin, Saxagliptin, Alogliptin, Teneligliptin, Anagliptin, and Evogliptin, A pharmaceutical composition for preventing or treating diabetic nephropathy, wherein the above biguanide drug is any one selected from the group consisting of metformin, buformin, and phenformin.

7. In the first paragraph, the pharmaceutical composition is a pharmaceutical composition for preventing or treating diabetic nephropathy, which exhibits at least one effect selected from the group consisting of the following i) to iii): i) Reduction of albuminuria; ii) reduction in the albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

8. In paragraph 1, the pharmaceutical composition is administered simultaneously, separately or sequentially with an antihypertensive agent and / or an antidiabetic agent, The above antihypertensive agent is any one selected from the group consisting of angiotensin Ⅱ receptor blocker and angiotensin-converting-enzyme inhibitor; A pharmaceutical composition for preventing or treating diabetic nephropathy, wherein the antidiabetic agent is any one selected from the group consisting of a sodium glucose cotransporter 2 inhibitor, a dipeptidyl peptidase-4 inhibitor, and a biguanide drug.

9. Zinc salt; and A pharmaceutical composition for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy, comprising cyclo-hispro or a pharmaceutically acceptable salt thereof as an active ingredient.

10. In paragraph 9, the antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, fimasartan, irbesartan, eprosartan, and azilsartan; or A pharmaceutical composition for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent for diabetic nephropathy, wherein the antihypertensive and / or antidiabetic agent is an angiotensin converting enzyme inhibitor selected from the group consisting of Captopril, Zofenopril, Enalapril, Ramipril, Quinapril, Perindopril, Lisinopril, Benazepril, Imidapril, Trandolapril, Cilazapril and Fosinopril.

11. In claim 9, the antidiabetic agent is a sodium glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin; A dipeptidyl peptidase-4 inhibitor selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, anagliptin, and evogliptin; or A pharmaceutical composition for enhancing the therapeutic effect of an antihypertensive and / or antidiabetic agent, wherein the antihypertensive agent is a biguanide drug selected from the group consisting of metformin, buformin and phenformin.

12. In the 9th paragraph, the pharmaceutical composition is a pharmaceutical composition for improving the treatment effect of an antihypertensive and / or antidiabetic agent for diabetic nephropathy, which exhibits at least one effect selected from the group consisting of the following i) to iii): i) Reduction of albuminuria; ii) reduction in the albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

13. In paragraph 9, the pharmaceutical composition is administered simultaneously, separately or sequentially with an antihypertensive agent and / or an antidiabetic agent, The above antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, fimasartan, 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, cilazapril, and fosinopril. The above antidiabetic agent is a sodium glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin; A pharmaceutical composition for enhancing the therapeutic effect of an antihypertensive and / or antidiabetic agent in diabetic nephropathy, wherein the antihypertensive and / or antidiabetic agent is a dipeptidyl peptidase-4 inhibitor selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, anagliptin, and evogliptin; or a biguanide drug selected from the group consisting of metformin, buformin, and phenformin.

14. Zinc salt; and A health functional food composition for preventing or improving diabetic nephropathy, comprising cyclo-hispro or a food-related acceptable salt thereof as an active ingredient.

15. In paragraph 14, the health functional food composition is for improving the diabetic nephropathy improvement effect in diabetic nephropathy patients receiving antihypertensive and / or antidiabetic agents, The above antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, fimasartan, 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, cilazapril, and fosinopril. The above antidiabetic agent is a sodium glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin; A health functional food composition for preventing or improving diabetic nephropathy, comprising a dipeptidyl peptidase-4 inhibitor selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, anagliptin, and evogliptin; or a biguanide drug selected from the group consisting of metformin, buformin, and phenformin.

16. In the 14th paragraph, the health functional food composition is a health functional food composition for preventing or improving diabetic nephropathy, which exhibits at least one effect selected from the group consisting of the following i) to iii): i) Reduction of albuminuria; ii) reduction in the albuminuria / creatinine ratio; and iii) Increased glomerular filtration rate.

17. In paragraph 14, the health functional food composition is administered simultaneously, separately, or sequentially with an antihypertensive agent and / or an antidiabetic agent, The above antihypertensive agent is an angiotensin II receptor blocker selected from the group consisting of losartan, valsartan, candesartan, olmesartan, telmisartan, fimasartan, 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, cilazapril, and fosinopril. The above antidiabetic agent is a sodium glucose cotransporter-2 inhibitor selected from the group consisting of dapagliflozin, canagliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, remogliflozin, remogliflozin etabonate, and ertugliflozin; A health functional food composition for preventing or improving diabetic nephropathy, comprising a dipeptidyl peptidase-4 inhibitor selected from the group consisting of sitagliptin, linagliptin, vildagliptin, gemigliptin, saxagliptin, alogliptin, teneligliptin, anagliptin, and evogliptin; or a biguanide drug selected from the group consisting of metformin, buformin, and phenformin.

18. A method for preventing, improving or treating diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof.

19. A method for improving the therapeutic effect of an antihypertensive and / or antidiabetic agent in diabetic nephropathy, comprising administering to a subject in need thereof an effective amount of a composition comprising zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof.

20. Use of a composition comprising a zinc salt and cyclo-hispro or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing, improving or treating diabetic nephropathy.

21. Use of a composition comprising a zinc salt for the manufacture of a drug for preventing, improving or enhancing the effect of an antihypertensive and / or antidiabetic agent on diabetic nephropathy; and cyclo-hispro or a pharmaceutically acceptable salt thereof.

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