Combination formulation for preventing or treating vascular or valvular stenosis

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating vascular or valvular stenosis, comprising a statin-based drug and a gliptin-based drug. When the statin-based drug and the gliptin-based drug are co-administered according to the present invention, calcification is synergistically reduced in comparison to when each of the statin-based drug and the gliptin-based drug is administered alone, and thus the composition according to the present invention is very useful for vascular or valvular stenosis.

Description

Combination preparations for the prevention or treatment of vascular or valvular stenosis

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of vascular or valvular stenosis, comprising a statin drug and a gliptin drug. Furthermore, the present invention relates to a health functional food composition for the prevention or improvement of vascular or valvular stenosis, comprising a statin drug and a gliptin drug.

[0002] Aortic valve stenosis (AVS) is a condition in which the aortic valve, located where blood flows from the left ventricle to the aorta, does not open properly during left ventricular contraction. The pathophysiology of ARS is complex and involves multiple pathological processes, including lipid retention, oxidation, chronic inflammation, fibrosis, and calcification. Calcification of the aortic valve is also called calcific aortic valve disease (CAVD). Currently, there are no approved pharmacological treatments specifically for ARS or CAVD, and therefore the only treatment options are surgical Aortic Valve Implantation (SAVI) and non-surgical transcatheter Aortic Valve Implantation (TAVI).

[0003] Cardiovascular calcification contributes to the exacerbation of hypertension, heart failure, acute coronary syndrome, and valvular disease, leading to various complications. Calcification occurs independently or in combination with fibrosis. Numerous epidemiological studies have shown that vascular calcification independently increases mortality. Vascular calcification occurs through mechanisms similar to the bone formation program during normal fetal development or after fracture, and is activated in aging, diabetes, chronic renal failure, and chronic inflammatory diseases. Inflammatory responses accelerate mineral loss from bone, and the released mineral is phagocytosed by abnormal vascular endothelium. Vascular calcification leads to vascular stenosis. Meanwhile, calcification also occurs in heart valves, and calcification of the aortic valve, particularly the aortic valve, can lead to aortic valve stenosis.

[0004] The calcification process of the aortic valve, which leads to aortic stenosis, begins deep within the aortic valve tissue, near the margin of the insertion. In advanced disease, nodules extend beyond the surface of the stenotic valve. The initial stage of calcification is called aortic valve sclerosis, while the later stage, aortic stenosis, occurs when the functional valve surface is sufficiently reduced to cause blood flow impairment.

[0005] There is currently no noninvasive treatment available to cure CAVD.

[0006] However, US Patent No. 9,416,196 discloses that a DPP-4 (Dipeptidyl peptidase-4) inhibitor, such as gliptin, can treat aortic valve calcification. DPP-4 (Dipeptidyl peptidase-4), also known as CD26 (cluster of differentiation 26), is a protein known to be involved in immune regulation, apoptosis, and signal transduction. According to the US patent, when blood vessels and valves are calcified, the expression of DPP-4 increases, and when a DPP-4 inhibitor is administered, the calcification is significantly reduced, so it is disclosed that DPP-4 inhibitors can be usefully used for the treatment or prevention of blood vessels or valves. As DPP-4 inhibitors, several types of gliptin drugs have been disclosed, including sitagliptin, vildagliptin, saxagliptin, linagliptin, dutogliptin, gemigliptin, alogliptin, anagliptin, and evogliptin.

[0007] Meanwhile, statins are HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors, a general term for drugs widely used as treatments for cardiovascular diseases such as hyperlipidemia and hypertension.

[0008] Furthermore, it has been reported that statins inhibit cardiovascular calcification. For example, statin drugs such as atorvastatin have been reported to inhibit aortic smooth muscle calcification (Son et al., Circulation Research, vol. 98, 8, pp. 1024-1031).

[0009] However, a drug that can treat calcification of blood vessels or valves, particularly aortic valve calcification or aortic valve stenosis, has not yet been developed. As a result of the inventors' efforts to develop a drug for the prevention or treatment of the same, they discovered that when a statin and a gliptin are used together, there is a synergistic effect in suppressing calcification of blood vessels or valves compared to when a gliptin or statin is administered alone, thereby completing the present invention.

[0010] The present invention seeks to provide a new composition and method for the prevention or treatment of vascular or valve stenosis.

[0011] To achieve the above purpose,

[0012] One aspect of the present invention provides a pharmaceutical composition for preventing or treating vascular or valvular stenosis, comprising a statin drug and a gliptin drug.

[0013] Another aspect of the present invention provides a health functional food composition for preventing or improving vascular or valvular stenosis, comprising a statin drug or a gliptin drug.

[0014] The pharmaceutical composition according to the present invention can prevent or treat vascular or valvular stenosis.

[0015] Figure 1a is a graph of weight change after co-administration of evogliptin and a statin, Figure 1b is a graph of weight change after co-administration of linagliptin and a statin, Figure 1c is a graph of weight change after co-administration of saxagliptin and a statin, and Figure 1d is a graph of weight change after co-administration of sitagliptin and a statin.

[0016] Figure 2a is a graph of plasma calcium levels after co-administration of evogliptin and a statin, Figure 2b is a graph of plasma calcium levels after co-administration of linagliptin and a statin, Figure 2c is a graph of plasma calcium levels after co-administration of saxagliptin and a statin, and Figure 2d is a graph of plasma calcium levels after co-administration of sitagliptin and a statin.

[0017] Figure 3a is a graph of plasma phosphate levels after co-administration of evogliptin and a statin, Figure 3b is a graph of plasma phosphate levels after co-administration of linagliptin and a statin, Figure 3c is a graph of plasma phosphate levels after co-administration of saxagliptin and a statin, and Figure 3d is a graph of plasma phosphate levels after co-administration of sitagliptin and a statin.

[0018] Figures 4a, 4b, and 4c are histopathological profiles of samples administered with evogliptin and statins together. (Scale bar = 100 μm, A = G1, B = G2, C = G3, D = G4, E = G8, F = G9, G = G10, H = G11, I = G12, J = G13, K = G14, L = G15, M = G16, N = G17)

[0019] Figures 5a, 5b, and 5c are histopathological profiles of samples co-administered with linagliptin and statins. (Scale bar = 100 μm, A = G1, B = G2, C = G3, D = G5, E = G8, F = G9, G = G10, H = G11, I = G12, J = G18, K = G19, L = G20, M = G21, N = G22)

[0020] Figures 6a, 6b, and 6c are histopathological profiles of samples administered with saxagliptin and statins. (Scale bar = 100 μm, A = G1, B = G2, C = G3, D = G6, E = G8, F = G9, G = G10, H = G11, I = G12, J = G23, K = G24, L = G25, M = G26, N = G27)

[0021] Figures 7a, 7b, and 7c are histopathological profiles of samples co-administered with sitagliptin and statins. (Scale bar = 100 μm, A = G1, B = G2, C = G3, D = G7, E = G8, F = G9, G = G10, H = G11, I = G12, J = G28, K = G29, L = G30, M = G31, N = G32)

[0022] Figure 8 is a graph showing the degree of aortic valve calcification in samples administered with statin and gliptin drugs alone.

[0023] Figures 9a and 9b are graphs showing the degree of aortic valve calcification in samples administered with linagliptin and lovastatin.

[0024] Figure 10 is a graph showing the degree of aortic valve calcification in samples administered with evogliptin and lovastatin.

[0025] Figure 11 is a graph showing the degree of aortic valve calcification in samples administered with evogliptin and rosuvastatin.

[0026] Figure 12 is a graph showing the degree of aortic valve calcification in samples administered with sitagliptin and rosuvastatin.

[0027] Figure 13 is a graph showing the degree of aortic valve calcification in samples administered with sitagliptin and lovastatin.

[0028] Figure 14 is a graph showing the degree of aortic valve calcification in samples administered with saxagliptin and lovastatin.

[0029] Figure 15 is a graph showing the degree of aortic valve calcification in samples administered with linagliptin and simvastatin together.

[0030] Figure 16 is a graph showing the degree of aortic valve calcification in samples administered with sitagliptin and atorvastatin.

[0031] Figures 17a and 17b are graphs of descending aortic calcium levels in samples co-administered with linagliptin and lovastatin, and Figures 17c and 17d are graphs of renal calcium levels in samples co-administered with linagliptin and lovastatin.

[0032] Figures 18a and 18b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with evogliptin and lovastatin.

[0033] Figures 19a and 19b are graphs of descending aorta and renal calcium levels, respectively, of samples co-administered with evogliptin and rosuvastatin.

[0034] Figures 20a and 20b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with sitagliptin and rosuvastatin.

[0035] Figures 21a and 21b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with sitagliptin and lovastatin in combination.

[0036] Figures 22a and 22b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with saxagliptin and lovastatin in combination.

[0037] Figures 23a and 23b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with linagliptin and simvastatin together.

[0038] Figures 24a and 24b are graphs of descending aorta and renal calcium levels, respectively, of samples administered with sitagliptin and atorvastatin together.

[0039] Hereinafter, the present invention will be described in detail.

[0040] The embodiments of the present invention may be modified in various ways, and the scope of the present invention is not limited to the embodiments described below. Furthermore, the embodiments of the present invention are provided to more fully explain the present invention to those of ordinary skill in the art.

[0041] Furthermore, reference to an element "including" throughout the specification does not exclude other elements, but rather includes other elements, unless specifically stated otherwise.

[0042]

[0043] One aspect of the present invention provides a combination of a statin drug and a gliptin drug for preventing or treating vascular or valvular stenosis.

[0044] One aspect of the present invention provides a pharmaceutical composition for preventing or treating vascular or valvular stenosis, comprising a statin drug and a gliptin drug.

[0045] Another aspect of the present invention provides a health functional food composition for preventing or improving valvular stenosis, comprising a statin drug or a gliptin drug.

[0046] In another aspect of the present invention, a method for preventing or treating vascular or valvular stenosis is provided by administering a statin drug and a gliptin drug to a subject in need of prevention or treatment of vascular or valvular stenosis in an amount effective for prevention or treatment.

[0047] In another aspect of the present invention, the use of a combination comprising a statin drug and a gliptin drug is provided for the manufacture of a medicament for preventing or treating vascular or valvular stenosis.

[0048] In another aspect of the present invention, a combination comprising the statin drug and the gliptin drug is provided for use in preventing or treating vascular or valvular stenosis.

[0049] In another aspect of the present invention, a combination formulation comprising the statin drug and the gliptin drug is provided for use in preventing or treating vascular or valvular stenosis.

[0050]

[0051] Throughout this specification, the term "combination" refers to a clinical practice involving the simultaneous or concurrent administration of a statin and a gliptin for the purpose of preventing or treating vascular or valvular stenosis. The compositions, methods, uses, combinations, and combination preparations described herein all presuppose combination use. Specific details regarding possible combinations are described below.

[0052]

[0053] The following description applies to all forms of the present invention, including the compositions, methods, uses, combinations and combination preparations.

[0054] The above blood vessel includes all blood vessels in the body, more specifically, the cardiovascular system related to the heart, and more specifically, may include the aorta, vena cava, pulmonary artery, vena cava, and renal artery. The above valve is a heart valve, and includes the aortic valve, pulmonary valve, mitral valve, and tricuspid valve, but preferably refers to the aortic valve. The above stenosis may occur in patients suffering from cardiovascular diseases such as heart failure and arteriosclerosis, or patients suffering from renal diseases such as chronic renal failure.

[0055] The above vascular or valvular stenosis may be accompanied by calcification of the vessel or valve. Vascular calcification occurs in patients with cardiovascular diseases such as heart failure, arteriosclerosis, or atherosclerosis, or in patients with renal diseases such as renal artery atherosclerosis or renal artery atherosclerosis. Renal diseases include chronic kidney disease.

[0056] Vascular calcification or stenosis, which occurs in patients with cardiovascular or kidney disease, can occur in any blood vessel in the body.

[0057] In the present invention, calcification refers to a condition in which calcium deposits in the heart, lungs, kidneys, blood vessels, or valves harden the tissues. As calcification progresses, kidney calcification can lead to kidney disease, blood vessels or valves can narrow, and aortic valve calcification can lead to aortic stenosis.

[0058] Calcification includes medial calcification or atherosclerotic calcification. Calcified tissue is called calcificum. Intimal calcification occurs in association with atherosclerosis. Atherosclerosis initially begins with the accumulation of lipid-rich macrophages and T lymphocytes on the inner surface of blood vessels or valves, forming a fatty layer. This is followed by the migration of smooth muscle cells from the media. The chemodynamic factors that stimulate this migration are thought to be produced by the adjacent endothelium, activated phagocytes, and other sources.

[0059] The migrated smooth muscle cells proliferate, accumulate fat, and produce extracellular matrix. Calcification occurs at the center of the atherosclerotic plaque.

[0060] Medial calcification occurs independently of atherosclerosis and intimal calcification. Medial calcification in distal arteries, also known as Mückeberg's sclerosis, is commonly observed in elderly diabetic patients. Its development is known to involve smooth muscle cells and elastin.

[0061] It has been reported that calcification caused by renal dysfunction is mainly medial calcification that occurs in the media, while calcification caused by atherosclerosis or calcification related to atherosclerosis is mainly reported to occur in the intima, which is a difference.

[0062] In addition, in patients with kidney disease, more specifically, patients with chronic renal failure or chronic kidney disease, calcification of the kidney itself occurs, and thus, chronic kidney disease, more specifically, chronic kidney disease accompanied by calcification, can be alleviated or treated through the composition of the present invention.

[0063]

[0064] The treatable stenosis in the present invention is not particularly limited, but includes those that are clinically diagnosed as stenosis, and may include an early stage of hardening that is not clinically diagnosed as stenosis. It may include a stage in which symptoms of fibrosis and / or calcification are confirmed, and preferably, the stenosis may include a stage in which symptoms of calcification are confirmed. The term “treatment” in the present invention includes slowing down the progression of stenosis or returning stenosis symptoms to a normal state by alleviating the degree of stenosis in the target tissue. Slowing down the progression of stenosis is a relative concept, and means significantly slowing down the rate of stenosis progression compared to when the combination treatment according to the present invention is not performed, for example, when no treatment is performed or only a general statin drug or gliptin drug is administered alone. When we say that the progression of stenosis is slowed, it can mean slowing it down by, for example, 1% or more, 2% or more, 3% or more, 5%, 10%, 15%, 20%, 30%, 40%, or 50%, and the degree of stenosis progression can be quantitatively evaluated through, for example, blood flow velocity, degree of calcification, etc.

[0065] When stenosis is accompanied by calcification, it is a concept that includes not only making the calcification completely normal by returning it to normal tissue, but also alleviation such as slowing down the progression of calcification or reducing the degree of calcification that has already occurred. Slowing down the progression of calcification is a relative concept, and means significantly slowing down the rate of progression of calcification compared to when the combination treatment according to the present invention is not performed, specifically compared to when no treatment is performed or only statin or gliptin is treated alone. Slowing down the progression of calcification includes, for example, slowing down the rate of increase in the amount of calcium deposited as an indicator of calcification or the area or weight of calcified tissue by 1% or more, 2% or more, 3% or more, 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, or 50% or more. When it is said that the degree of calcification is reduced, it includes reducing the area of ​​calcified tissue by 1% or more, 2% or more, 3% or more, 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, or 50% or more, based on the area of ​​calcified tissue before starting the combined administration of the statin drug and the gliptin drug according to the present invention.

[0066] As a method for quantifying the area of ​​calcified tissue, a method for quantifying the area observed by staining with hematoxylin and eosin (HE) for histopathology or von Kossa (VK) for calcium deposition can be used.

[0067] “Prevention” in the present invention may include alleviating calcification before calcification progresses or preventing worsening during calcification progress.

[0068]

[0069] In the present invention, the statin drug is not specifically limited to an HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitor, but may include at least one statin drug selected from fluvastatin, atorvastatin, lovastatin, simvastatin, and rosuvastatin, and the gliptin drug is not specifically limited to a DPP-4 inhibitor, but may include at least one gliptin drug selected from evogliptin, linagliptin, saxagliptin, and sitagliptin.

[0070] In the present invention, statin drugs and gliptin drugs may be included in the composition or administered in a therapeutically effective amount capable of preventing calcification of blood vessels or valves. The pharmaceutical composition or health functional food composition according to the present invention includes not only a single formulation for administration in which the statin drug and gliptin drug are physically mixed, but also a formulation prepared for separate administration and then administered together.

[0071] It is desirable that the dosage of statin drugs and gliptin drugs be determined so that they are administered in a therapeutically effective amount within a range that is not harmful to the subject.

[0072] And, the dosage of these drugs may vary depending on the patient's condition, weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease, and a person skilled in the art can determine an appropriate amount. For example, each of the statin drugs and the gliptin drugs may be administered at a dosage of 0.01 to 100 mg / kg per day, and may be administered once or several times a day, for example, twice, three times, or four times.

[0073] Furthermore, their dosing cycle can be determined daily, on a 2-7-day cycle, or irregularly. The dosing cycle can range from 2 days to 1 month in the short term, from more than 1 month to 3 months in the medium term, from more than 3 months to 1-3 years in the long term, or even throughout the remaining lifespan. It is clear that the dosage and dosing cycle can vary depending on the number of dosing cycles.

[0074] The dosage ratio of the statin drug and the gliptin drug can be determined by a person skilled in the art as an amount effective for treatment, and for example, can be determined within the range of 1:150 to 150:1, 1:100 to 100:1, 1:50 to 50:1, 1:10 to 10:1, 15:1 to 1:1, and 10:1 to 1:1 based on the weight of the active ingredient.

[0075] The statin drug may be administered in a concentration range of 0.05 mg / kg to 50 mg / kg, 0.1 mg / kg to 40 mg / kg, 0.1 mg / kg to 30 mg / kg, preferably 0.125 mg / kg to 20 mg / kg, and the gliptin drug may be administered in a concentration range of 0.5 mg / kg to 50 mg / kg, 1 mg / kg to 40 mg / kg, 1 mg / kg to 30 mg / kg, preferably 1 mg / kg to 20 mg / kg.

[0076] In the present invention, the statin drug and the gliptin drug may be administered simultaneously or concurrently. When referring to simultaneous administration, it does not only include co-administration in a physically mixed form, but also concurrent administration within a time gap of 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, or 1 hour in which drug interaction is possible. The pharmaceutical composition or health functional food composition according to the present invention includes not only a single formulation in which the statin drug and the gliptin drug are physically mixed, but also a formulation provided as separate formulations and administered concurrently. When provided as separate formulations, they may be administered simultaneously or concurrently as described above. The term "pharmaceutical composition" in the present invention is not limited to a physically mixed composition, but is intended to indicate use as a medicine.

[0077] Even if administered at different times, the effect of combined drug administration can be achieved by adjusting the blood drug profile so that the statins and gliptins in the blood can interact, and for example, they can be administered at different times in the range of more than 6 hours to within 48 hours.

[0078] For combination therapy, statin and gliptin drugs can be formulated independently. Commercially available formulations containing each drug as an active ingredient, targeting vascular or valvular stenosis as well as other indications, can also be used as-is.

[0079]

[0080] For combined administration, the active ingredients of statin drugs and gliptin drugs can be formulated together as a combination drug in a single formulation.

[0081] When formulated independently or as a combination formulation, the formulation may be formulated for oral administration, such as tablets, capsules, powders, granules, or suspensions, or for parenteral administration, such as injections, but is preferably formulated for oral administration. When formulated, these formulations may further include one or more pharmaceutically acceptable additives. These additives may include one or more selected from the group consisting of excipients (diluents), binders, disintegrants, lubricants, and colorants.

[0082]

[0083] The composition according to the present invention may be used alone or in combination with methods using surgery, hormone therapy, drug therapy and biological response modifiers for the prevention or treatment of vascular or valve calcification.

[0084] For combined administration, a health functional food composition containing both the active ingredient of a statin drug and the active ingredient of a gliptin drug can be formulated together.

[0085] There are no particular limitations on other ingredients that may be included in the above health functional food composition, and, like conventional foods, it may contain various herbal extracts, food additives, or natural carbohydrates as additional ingredients. In addition, food additives may be additionally included, and food additives include flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like, which are conventionally used in the art.

[0086] It is also clear that in health functional food compositions, statin drugs and gliptin drugs are provided as separate formulations and then used together.

[0087] For the above health functional food composition, the dosage, administration cycle, and administration days can all be applied as described above.

[0088] Hereinafter, the present invention will be described in detail through examples.

[0089] However, the examples described below are only specific examples of one aspect of the present invention, and the present invention is not limited thereto.

[0090]

[0091] <Experimental Preparation> Preparations for Animal Testing

[0092] 1. Prepare warfarin and vitamin K1 solution

[0093] Dissolve the exact weight of warfarin sodium and vitamin K1 in saline solution, and dilute appropriately with saline solution to make 7.57×10 each. -4 Warfarin solution with a concentration of mol / kg / 100μl and 5.32×10 -5 Vitamin K was obtained at a concentration of mol / kg / 100μl.

[0094] 2. Prepare vitamin D3 solution

[0095] 6.5X10 of exact weight of vitamin D3 in a mixture of 30% PEG400, 0.5% Tween80, 5% Propylene glycol, and 64.5% saline solution 4 A vitamin D3 solution was obtained by dissolving IU / ml (1.625 mg / ml) using sonication for 1 hour.

[0096] 3. Preparation of gliptin and statin

[0097] Accurately weigh all ingredients and dissolve or disperse in 5% CMC sodium salt to prepare evogliptin 1 mg / kg / 100 μl, linagliptin 1 mg / kg / 100 μl, saxagliptin 1 mg / kg / 100 μl, sitagliptin 10 mg / kg / 100 μl, fluvastatin 8 mg / kg / 100 μl, atorvastatin 8 mg / kg / 100 μl, lovastatin 8 mg / kg / 100 μl, simvastatin 8 mg / kg / 100 μl, and rosuvastatin 4 mg / kg / 100 μl.

[0098] 4. Prepare the animal (mouse)

[0099] Animals: C57BL / 6 mice (OrientBio, Seongnam, Korea)

[0100] Gender: Male

[0101] Age on arrival: C57BL / 6 (6 weeks)

[0102] Housing: Animals were housed in filter-capped polycarbonate cages, five per cage, in a sterile cleanroom with controlled temperature (25°C) and humidity (45–55%). The light:dark cycle was 12 h:12 h, and sterilized food and water were provided ad libitum.

[0103] Guidelines: For the warfarin model, animals were handled according to the Animal Care and Handling Guidelines of Yeungnam University, Gyeongsan, Republic of Korea, and for the vitamin D model, animals were handled according to the Animal Care and Handling Guidelines of Daegu Haany University, Gyeongsan, Republic of Korea.

[0104]

[0105] <Experimental Example 1> Experimental Method 1: In vivo drug administration

[0106] Mouse model of calcification induced by warfarin administration

[0107] After acclimation for one week, the mice were randomly divided into 32 groups.

[0108] All groups, except the untreated control group (G1), received both warfarin and vitamin K1 at 8:00 AM and warfarin alone at 8:00 PM for 4 weeks.

[0109] Simultaneously, G3 was orally administered 5% CMC (Carboxymethyl Cellulose), and G4, G5, G6, G7, G8, G9, G10, G11, and G12 were orally administered evogliptin (1 mg / kg, G4), linagliptin (1 mg / kg, G5), saxagliptin (1 mg / kg, G6), sitagliptin (10 mg / kg, G7), fluvastatin (8 mg / kg, G8), atorvastatin (8 mg / kg, G9), lovastatin (8 mg / kg, G10), simvastatin (8 mg / kg, G11), and rosuvastatin (4 mg / kg, G12) at 8:00 AM for 4 weeks, respectively. In contrast, groups G13 to G32 were orally administered a combination of gliptin and statin at 8:00 AM for 4 weeks. Four weeks later, whole blood and intact hearts were excised from all mice under anesthesia. Plasma was separated from the blood to measure plasma calcium and phosphate levels. The excised hearts were preserved in buffered formalin until further histopathological and histomorphological examination.

[0110] Each group was treated as shown in Table 1 below.

[0111] Five to eight heart tissue samples were obtained from each group and used for analysis.

[0112] Warfarin + Vitamin K1 Treatment Drug Administration G1 Not Treated G2 Not Treated G3 Treatment Vehicle G4 Treatment Evogliptin 1 mg / kg G5 Treatment Linagliptin 1 mg / kg G6 Treatment Saxagliptin 1 mg / kg G7 Treatment Sitagliptin 10 mg / kg G8 Treatment Fluvastatin 8 mg / kg G9 Treatment Atorvastatin 8 mg / kg G10 Treatment Lovastatin 8 mg / kg G11 Treatment Simvastatin 8 mg / kg G12 Treatment Rosuvastatin 4 mg / kg G13 Treatment Evogliptin 1 mg / kg and Fluvastatin 8 mg / kg G14 Treatment Evogliptin 1 mg / kg and Atorvastatin 8 mg / kg G15 Treatment Evogliptin 1 mg / kg and Lovastatin 8 mg / kg G16 Treatment Evogliptin 1 mg / kg and simvastatin 8 mg / kgG17 Treatment with evogliptin 1 mg / kg and rosuvastatin 4 mg / kgG18 Treatment with linagliptin 1 mg / kg and fluvastatin 8 mg / kgG19 Treatment with linagliptin 1 mg / kg and atorvastatin 8 mg / kgG20 Treatment with linagliptin 1 mg / kg and lovastatin 8 mg / kgG21 Treatment with linagliptin 1 mg / kg and simvastatin 8 mg / kgG22 Treatment with linagliptin 1 mg / kg and rosuvastatin 4 mg / kgG23 Treatment with saxagliptin 1 mg / kg and fluvastatin 8 mg / kgG24 Treatment with saxagliptin 1 mg / kg and atorvastatin 8 mg / kgG25 Treatment with saxagliptin 1 mg / kg and lovastatin 8 mg / kgG26 Treatment with saxagliptin 1 mg / kg and simvastatin 8 mg / kgG27 Treatment with saxagliptin 1 mg / kg and rosuvastatin 4 mg / kgG28 Treatment with sitagliptin 10 mg / kg and fluvastatin 8 mg / kgG29 Treatment with sitagliptin 10 mg / kg and atorvastatin 8 mg / kgG30 Treatment with sitagliptin 10 mg / kg and lovastatin 8 mg / kgG31 Treatment with sitagliptin 10 mg / kg and simvastatin 8 mg / kgG32 Treatment with sitagliptin 10 mg / kg and rosuvastatin 4 mg / kg

[0113] result

[0114] 1. Weight changes

[0115] In all groups, including control and CAVD-induced C57BL / 6 mice, administration of gliptin and statins alone or in combination resulted in a gradual and slight increase in body weight over time (Fig. 1).

[0116] 2. Plasma calcium levels

[0117] G17 (evogliptin + rosuvastatin) significantly reduced plasma calcium levels compared to G1 (control) (Figure 2). However, no significant differences were found between the gliptin or statin monotherapy groups and the gliptin / statin combination therapy groups.

[0118] 3. Plasma phosphate levels

[0119] After 4 weeks of treatment, the plasma phosphate concentration changes were measured, and the groups receiving fluvastatin 8 mg / kg (G8), evogliptin 1 mg / kg and fluvastatin 8 mg / kg (G13), evogliptin 1 mg / kg and simvastatin 8 mg / kg (G16), evogliptin 1 mg / kg and rosuvastatin 8 mg / kg (G17), and linagliptin 1 mg / kg and atorvastatin 8 mg / kg (G19) showed differences compared to G3 (vehicle) (Fig. 3). However, no significant differences were found between the gliptin or statin monotherapy groups and the gliptin / statin combination therapy groups.

[0120] 4. Heart tissue analysis

[0121] 1) Histopathological examination method

[0122] Individual heart samples were cross-sectioned around the origin / root of the ascending aorta. Histological sections were then prepared from all 245 samples. All cross-sectioned cardio-aortic valve areas were refixed in 10% neutral buffered formalin for 24 hours to prepare histological samples. After paraffin embedding, two serial sections, 3–4 μm thick, were prepared from each paraffin block and stained with hematoxylin and eosin (HE) for routine histopathology or von Kossa (VK) for calcium deposition. To observe changes in more detail, a computer-assisted image analysis program and a histological camera system were used to calculate the mean aortic valve thickness (μm) and the number of inflammatory cells infiltrating the aortic valve area (cells / mm2) in the HE staining, and the mean calcified aortic valve area (% / mm2) in the VK staining in the histopathological analysis. The histopathologist was blinded to group distribution when this analysis was performed. Histopathological examinations of one section of each aortic valve, cross-sections, histological sections of each aortic valve, and a total of 5–8 C57BL / 6 mouse heart samples from each group were statistically analyzed.

[0123] 2) Summary of analysis results

[0124] Co-administration of warfarin / vitamin K1 to C57BL / 6 mice induced typical CAVD-related histopathological changes, such as marked aortic valve thickening due to inflammatory cell infiltration and calcium deposition, compared with the intact control (untreated, G1) C57BL / 6 mouse heart-aortic valve site, which was reconfirmed by histomorphometric analysis (Figs. 4a to 7c); the mean aortic valve thickness, the number of inflammatory cells infiltrating the aortic valve site, and the calcified aortic valve site in the control C57BL / 6 mice (G2 and G3) not administered warfarin / vitamin K1 were observed to be significantly increased, respectively, compared with the normal control C57BL / 6 mouse heart-aortic valve site (G1). However, these histopathological signs of CAVD induced by warfarin / vitamin K1 were significantly suppressed by single-agent administration of four types of gliptins and five types of statins, and were confirmed to be consistent with the case of combined administration of 20 types compared to the control C57BL / 6 mouse cardio-aortic valve area (G3) administered with warfarin / vitamin K1, at least under the current histopathological analysis conditions. In particular, co-administration of evogliptin 1 mg / kg and lovastatin 8 mg / kg (G15), evogliptin 1 mg / kg and rosuvastatin 4 mg / kg (G17), linagliptin 1 mg / kg and lovastatin 8 mg / kg (G20), linagliptin 1 mg / kg and simvastatin 8 mg / kg (G21), saxagliptin 1 mg / kg and lovastatin 8 mg / kg (G25), sitagliptin 10 mg / kg and atorvastatin 8 mg / kg (G29), sitagliptin 10 mg / kg and lovastatin 8 mg / kg (G30), and sitagliptin 10 mg / kg and rosuvastatin 4 mg / kg (G32) was shown to have a significantly higher toxicity than the same gliptins (G4 to G7) at least under these histopathological analysis conditions. Compared with single-agent administration of statins (G8-12), warfarin / vitamin K1 showed a significantly more favorable inhibitory effect on CAVD histopathological signs.

[0125] 3) Results of combined administration of evogliptin and statins

[0126] Index (unit) Group average thickness (μm) Number of inflammatory cells (cells / mm 2 )Calcification area (% / mm) 2 )G143.04±6.3530.40±11.260.40±0.22G2135.04±10.64 a 428.00±83.77 a 10.92±0.93 a G3137.21±12.38 a 425.60±60.12 a 11.44±1.44 a G483.44±10.93 ace 169.00±27.15 ade 3.39±0.44 ace G890.53±7.31 acf 172.75±32.16 ade 6.46±1.05 acf G981.22±8.89 ace 180.50±23.56 adf 4.62±0.90 ace G1089.44±7.82 ace 162.50±11.51 adf 5.17±1.12 ace G1197.78±9.72 adf 186.75±20.06 adf 6.04±1.10 ace G1288.86±12.09 ace 145.75±19.23 ade 5.78±1.55 ace G1359.39±3.86 cehi 78.25±15.65 adegi 2.46±0.62 acei G1483.26±13.28 ace 182.50±27.64 ade 3.40±1.13 ace G1553.35±4.36 cegj 66.50±18.85 degj 1.36±0.48 bcegk G1684.49±14.71 bce 171.00±30.22ade 4.96±1.48 bce G1760.69±6.80 bcehl 49.50±13.80 degl 1.18±0.47 cegi

[0127] Data values ​​are expressed as the mean ± standard deviation of eight cardio-aortic valve tissue fields, excluding G1, G2, and G3, which analyzed five cardio-aortic valve tissue fields, and eight cardio-aortic valve tissue fields, excluding G16, which analyzed six cardio-aortic valve tissue fields. a p<0.01 and b p<0.05, compared with G1 by DT3 test; c p<0.01 and d p<0.05, compared with G2 by DT3 test; e p<0.01 and f p<0.05, compared with G3 by DT3 test; g p<0.01 and h Compared with G4 by DT3 test at p<0.05; i Compared to G8 by DT3 test with p<0.01; j p<0.01 and k Compared with G10 by DT3 test at p<0.05; l Compared to G12 by DT3 test with p<0.01.

[0128] 4) Results of combined administration of linagliptin and statins

[0129] Index (unit) Group average thickness (μm) Number of inflammatory cells (cells / mm 2 )Calcification area (% / mm) 2 )G143.04±6.3530.40±11.260.40±0.22G2135.04±10.64 a 428.00±83.77 j 10.92±0.93 j G3137.21±12.38 a 425.60±60.12 j 11.44±1.44 j G585.61±7.46acd 184.00±20.95 jmo 4.92±0.92 jln G890.53±7.31 acd 172.75±32.16 jmn 6.46±1.05 jlo G981.22±8.89 acd 180.50±23.56 jmo 4.62±0.90 jln G1089.44±7.82 acd 162.50±11.51 jmo 5.17±1.12 jln G1197.78±9.72 acd 186.75±20.06 jmo 6.04±1.10 jln G1288.86±12.09 acd 145.75±19.23 jmn 5.78±1.55 jln G1865.99±7.90 bcdfg 86.75±17.76 jmnpq 3.27±0.75 jlnq G1967.34±5.41 acdf 120.75±17.27 jmnpr 4.27±0.63 jln G2054.40±10.36 cdfh 53.75±18.93 lnps 0.80±0.43 lnps G2157.41±8.00 cdfi 67.00±16.53 kmnpt 2.19±0.52 jlnpt G2285.80±19.37 acd 178.63±25.81 jmn 4.94±1.33 jln

[0130] Data values ​​are presented as mean ± standard deviation of eight cardiac aortic valve histological fields, excluding G1, G2, and G3, in which five cardiac aortic valve histological fields were analyzed. a p<0.01 and b p<0.05 compared to G1 by THSD test, cp<0.01 compared to G2 by THSD test, d p<0.01 compared to G3 by THSD test, e p<0.01 and f p<0.05 compared to G5 by DT3 test; g p<0.01, compared to G8 by THSD test, h p<0.01, compared to G10 by THSD test, i p<0.01, compared to G11 by THSD test, j p<0.01, k p<0.05, compared to G1 by DT3 test, l p<0.01, m p<0.05, compared to G2 by DT3 test, n p<0.01, o p<0.05, compared with G3 by DT3 test; p Compared with G5 by DT3 test with p<0.01; q Compared with G8 by DT3 test with p<0.01; r Compared with G9 by DT3 test with p<0.01; s Compared with G10 by DT3 test with p<0.01; t Compared to G11 by DT3 test with p<0.01.

[0131] 5) Results of combined administration of saxagliptin and statins

[0132] Index (unit) Group average thickness (μm) Number of inflammatory cells (cells / mm 2 )Calcification area (% / mm) 2 )G143.04±6.3530.40±11.260.40±0.22G2135.04±10.64 a 428.00±83.77 a 10.92±0.93 a G3137.21±12.38 a 425.60±60.12 a 11.44±1.44 a G674.74±4.65 ace128.75±21.25 ade 3.07±0.33 ace G890.53±7.31 acf 172.75±32.16 ade 6.46±1.05 acf G981.22±8.89 ace 180.50±23.56 adf 4.62±0.90 ace G1089.44±7.82 ace 162.50±11.51 adf 5.17±1.12 ace G1197.78±9.72 adf 186.75±20.06 adf 6.04±1.10 ace G1288.86±12.09 ace 145.75±19.23 ade 5.78±1.55 ace G2366.32±17.79 ce 170.88±24.19 ade 3.58±1.00 aceh G2475.43±12.32 ace 182.00±41.49 ade 3.19±0.88 ace G2554.85±5.05 cegi 46.25±11.44 degi 1.95±0.27 acegj G2670.14±12.12 bcek 173.00±12.24 adf 3.55±0.84 acek G2786.62±13.07 aceg 121.00±27.28 ade 3.24±0.83 ace

[0133] Data values ​​are presented as mean ± standard deviation of eight aortic valve tissue areas, excluding G1, G2, and G3, where five aortic valve tissues were analyzed. a p<0.01 and b p<0.05, compared with G1 by DT3 test; c p<0.01 and dp<0.05, compared with G2 by DT3 test; e p<0.01 and f p<0.05, compared with G3 by DT3 test; g Compared with G6 by DT3 test with p<0.01; h Compared to G8 by DT3 test with p<0.01; i p<0.01 and j Compared with G10 by DT3 test at p<0.05; k Compared to G11 by DT3 test with p<0.05.

[0134] 6) Results of combined administration of sitagliptin and statins

[0135] Index (unit) Group average thickness (μm) Number of inflammatory cells (cells / mm 2 )Calcification area (% / mm) 2 )G143.04±6.3530.40±11.260.40±0.22G2135.04±10.64 a 428.00±83.77 h 10.92±0.93 h G3137.21±12.38 a 425.60±60.12 h 11.44±1.44 h G789.76±13.22 abc 209.25±45.28 hm 4.59±1.06 hjl G890.53±7.31 abc 172.75±32.16 hkl 6.46±1.05 hjm G981.22±8.89 abc 180.50±23.56 hkm 4.62±0.90 hjl G1089.44±7.82 abc 162.50±11.51 hkm 5.17±1.12 hjl G1197.78±9.72 abc 186.75±20.06 hkm 6.04±1.10 hjl G1288.86±12.09 abc145.75±19.23 hkl 5.78±1.55 hjl G2881.95±16.25 abc 198.75±67.56 hkl 5.16±1.24 hjl G2955.94±11.80 bcde 60.00±11.76 klno 1.33±0.49 ijlno G3056.50±8.71 bcdf 69.75±12.58 iklnp 2.03±0.30 hjlnq G3180.31±7.31 abc 171.50±22.34 hkl 5.34±1.58 hjl G3254.37±12.37 bcdg 55.25±10.36 klnr 1.40±0.58 jlnr

[0136] Data values ​​are presented as mean ± SD of eight aortic valve tissue areas, excluding G1, G2, and G3, where five aortic valve tissues were analyzed. a p<0.01, compared with G1 by THSD test; b p<0.01, compared with G2 by THSD test; c p<0.01, compared with G3 by THSD test; d p<0.01, compared with G7 by DT3 test; e p<0.01, compared with G9 by THSD test; f p<0.01, compared with G10 by THSD test; g p<0.01, compared with G12 by THSD test; h p<0.01, i p<0.05, compared with G1 by DT3 test; j p<0.01, k p<0.05, compared with G2 by DT3 test; l p<0.01 and m p<0.05 compared to G3 by DT3 test, np<0.01 compared to G7 by DT3 test, o p<0.01 compared to G9 by DT3 test, p p<0.01 and q p<0.05 compared to G10 by DT3 test, r p<0.01 compared to G12.

[0137]

[0138] <Experimental Example 2> Experimental Method 2: In vivo drug administration

[0139] Mouse model of calcification induced by vitamin D administration

[0140] After acclimation for one week, the mice were randomly divided into groups of 12 to 18.

[0141] All groups except the untreated control group (Normal) received Vitamin D3 6.5X10 5 IU / kg was administered subcutaneously at 24-hour intervals for 3 days.

[0142] Simultaneously, group 2 was administered 0.5% CMC orally, and each gliptin and statin were administered orally at 1-hour intervals, individually or in combination, according to the concentration. Seven days later, the intact heart, kidney, and aorta were resected and isolated under anesthesia. The resected heart was preserved in buffered formalin until further histopathological and histomorphological examinations. In addition, the isolated left kidney and descending aorta were diluted 10 times the tissue volume with chilled saline, homogenized using a homogenizer, and stored in 0.6 N HCl at 4°C for 24 hours to separate calcium from the tissue. The supernatant separated using a refrigerated centrifuge was used to measure the amount of calcium. For accurate measurement, quantification was performed based on the amount of protein.

[0143] result

[0144] 1. Evaluation of the efficacy of statins and gliptins on aortic valve calcification, respectively.

[0145] Each statin and gliptin showed efficacy in inhibiting calcification in proportion to concentration (Fig. 8), and gliptin showed relatively higher efficacy than statins.

[0146] To confirm the combined effect of statins and gliptins, various combinations were considered, and there was no difference in the efficacy of statins and gliptins between the warfarin-induced CAVD and vitamin D-induced CAVD models, and since the degree of efficacy between each substance had a similar pattern, eight combinations were selected with reference to the efficacy results of the warfarin-induced CAVD mouse animal model, and the efficacy was evaluated in the vitamin D-induced CAVD mouse animal model. It is known that the vitamin D-induced CAVD mouse model shows a worse CAVD prognosis than the warfarin-induced CAVD mouse model.

[0147] 2. Aortic valve calcification analysis

[0148] The degree of calcification was analyzed by histopathological examination using the same method as in Experimental Example 1.

[0149] 1) Results of combined administration of linagliptin and lovastatin

[0150] All combinations of linagliptin at concentrations ranging from 0.125 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg demonstrated superior efficacy compared to single agents (Fig. 9). Furthermore, their efficacy was higher than the sum of the individual effects, demonstrating a synergistic effect.

[0151] 2) Results of combined administration of evogliptin and lovastatin

[0152] In all combinations, evogliptin was administered at concentrations ranging from 0.25 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg, demonstrating superior efficacy compared to either drug alone (Fig. 10). Synergistic effects were observed in all combinations, except for those administered at the maximum approved doses of evogliptin and lovastatin.

[0153] 3) Results of combined administration of evogliptin and rosuvastatin

[0154] In all combinations of evogliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and rosuvastatin at concentrations ranging from 1 mg / kg to 8 mg / kg, superior efficacy was observed compared to the efficacy of either drug alone (Fig. 11). Synergistic effects were observed in all combinations except for the combination at the maximum approved doses of evogliptin and rosuvastatin.

[0155] 4) Results of combined administration of sitagliptin and rosuvastatin

[0156] In all combinations of sitagliptin at concentrations ranging from 2.5 mg / kg to 20 mg / kg and rosuvastatin at concentrations ranging from 1 mg / kg to 8 mg / kg, superior efficacy was observed compared to the monotherapy efficacy (Fig. 12). Synergistic effects were observed in all combinations except for the combination at the maximum approved doses of sitagliptin and rosuvastatin.

[0157] 5) Results of combined administration of sitagliptin and lovastatin

[0158] In all combinations, ranging from 2.5 mg / kg to 20 mg / kg of sitagliptin and from 1 mg / kg to 12 mg / kg of lovastatin, superior efficacy was observed compared to the monotherapy (Fig. 13). Synergistic effects were observed in all combinations, except for those administered at the maximum approved doses of sitagliptin and lovastatin.

[0159] 6) Results of combined administration of saxagliptin and lovastatin

[0160] In all combinations, saxagliptin was administered at concentrations ranging from 0.25 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg, demonstrating superior efficacy compared to either drug alone (Fig. 14). Synergistic effects were observed in all combinations, except for those administered at the maximum approved doses of saxagliptin and lovastatin.

[0161] 7) Results of combined administration of linagliptin and simvastatin

[0162] In all combinations, linagliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and simvastatin at concentrations ranging from 1 mg / kg to 16 mg / kg, demonstrated superior efficacy compared to either drug alone (Fig. 15). Synergistic effects were observed in all combinations, including those administered at the maximum approved doses of linagliptin and simvastatin, respectively.

[0163] 8) Results of combined administration of sitagliptin and atorvastatin

[0164] In all combinations, ranging from 2.5 mg / kg to 20 mg / kg of sitagliptin and from 1 mg / kg to 16 mg / kg of atorvastatin, superior efficacy was observed compared to the monotherapy efficacy (Fig. 16). Synergistic effects were observed in all combinations, except for those administered at the maximum approved doses of sitagliptin and atorvastatin.

[0165] 3. Analysis of vascular calcification and renal calcification

[0166] The descending aorta and kidney were removed from mice at the end of the experiment. The left kidney was used. After removing any remaining blood within the tissue, the descending aorta and kidney were each pulverized using a tissue grinder. Calcium was measured using a calcium measurement kit, and the protein content of each tissue was quantified for quantification. The results were analyzed by correcting the amount of calcium (μg) to the amount of protein (mg).

[0167] 1) Results of combined administration of linagliptin and lovastatin

[0168] In all combinations of linagliptin at concentrations ranging from 0.125 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg, superior efficacy was observed compared to the single efficacy (Figs. 17a and 17b). Furthermore, the efficacy of these was higher than the sum of the individual efficacy values, showing that most of them exhibited a synergistic effect. When linagliptin was combined with 0.5 mg / kg and 12 mg / kg of lovastatin, an efficacy close to a synergistic effect was also observed.

[0169] 2) Results of combined administration of evogliptin and lovastatin

[0170] In all combinations, evogliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg, demonstrated superior efficacy compared to either alone (Figs. 18a and 18b). In the aorta, synergistic effects were observed in all combinations except for the maximum approved dose of evogliptin and lovastatin, and in the kidney, synergistic effects were observed in all combinations. This is consistent with the efficacy observed in CAVD.

[0171] 3) Results of combined administration of evogliptin and rosuvastatin

[0172] In all combinations, evogliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and rosuvastatin at concentrations ranging from 1 mg / kg to 8 mg / kg, demonstrated superior efficacy compared to either alone (Figures 19a and 19b). Synergistic effects were observed in all combinations, except for those administered at the maximum approved doses of evogliptin and rosuvastatin. This is consistent with the efficacy observed in CAVD.

[0173] 4) Results of combined administration of sitagliptin and rosuvastatin

[0174] In all combinations, ranging from 2.5 mg / kg to 20 mg / kg of sitagliptin and from 1 mg / kg to 8 mg / kg of rosuvastatin, superior efficacy was observed compared to the monotherapy (Figures 20a and 20b). Synergistic effects were observed in all combinations, except for the maximum approved dose of sitagliptin and rosuvastatin. This is consistent with the efficacy observed in CAVD.

[0175] 5) Results of combined administration of sitagliptin and lovastatin

[0176] All combinations of sitagliptin at concentrations ranging from 2.5 mg / kg to 20 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg demonstrated superior efficacy compared to either alone (Figures 21a and 21b). A synergistic effect was observed in all combinations except for the maximum approved dose of sitagliptin and lovastatin. This is consistent with the efficacy observed in CAVD.

[0177] 6) Results of combined administration of saxagliptin and lovastatin

[0178] In all combinations, saxagliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and lovastatin at concentrations ranging from 1 mg / kg to 12 mg / kg, demonstrated superior efficacy compared to either drug alone (Figures 22a and 22b). Synergistic effects were observed in all combinations, except when saxagliptin and lovastatin were administered at the maximum approved doses. This is consistent with the efficacy observed in CAVD.

[0179] 7) Results of combined administration of linagliptin and simvastatin

[0180] In all combinations, linagliptin at concentrations ranging from 0.25 mg / kg to 1 mg / kg and simvastatin at concentrations ranging from 1 mg / kg to 16 mg / kg, demonstrated superior efficacy compared to either drug alone (Figures 23a and 23b). Synergistic effects were observed in all combinations, including those administered at the maximum approved doses of linagliptin and simvastatin. This is consistent with the efficacy observed in CAVD.

[0181] 8) Results of combined administration of sitagliptin and atorvastatin

[0182] All combinations of sitagliptin at concentrations ranging from 2.5 mg / kg to 20 mg / kg and atorvastatin at concentrations ranging from 1 mg / kg to 16 mg / kg demonstrated superior efficacy compared to either alone (Figures 24a and 24b). For vascular calcification, a synergistic effect was observed in all combinations except for the maximum approved dose of sitagliptin and atorvastatin. For renal calcification, a synergistic effect was observed in all combinations.

Claims

1. A pharmaceutical composition for preventing or treating kidney disease or vascular or valvular stenosis, comprising a statin drug and a gliptin drug.

2. In paragraph 1, A pharmaceutical composition comprising the above valve, wherein the valve comprises an aortic valve.

3. In paragraph 1, A pharmaceutical composition wherein the above vascular or valve stenosis involves calcification of the blood vessel or valve, and the above renal disease involves calcification of the kidney.

4. In paragraph 3, A pharmaceutical composition wherein calcification of the blood vessels or valves is caused by arteriosclerosis or atherosclerosis.

5. In paragraph 3, A pharmaceutical composition wherein calcification of the blood vessels or kidneys occurs in patients with kidney disease.

6. In paragraph 1, A pharmaceutical composition, wherein the aortic valve stenosis includes calcific aortic valve disease (CAVD).

7. In paragraph 1, A pharmaceutical composition comprising at least one statin drug selected from the group consisting of fluvastatin, atorvastatin, lovastatin, simvastatin, and rosuvastatin.

8. In paragraph 1, A pharmaceutical composition comprising at least one drug selected from the group consisting of evogliptin, linagliptin, saxagliptin, and sitagliptin.

9. In paragraph 1, the statin drug is any one selected from the group consisting of fluvastatin, atorvastatin, lovastatin, simvastatin, and rosuvastatin. A pharmaceutical composition wherein the above gliptin series drug is any one selected from the group consisting of evogliptin, linagliptin, saxagliptin, and sitagliptin.

10. A pharmaceutical composition in paragraph 1, wherein the statin drug and the gliptin drug are administered in combination.

11. A pharmaceutical composition according to claim 1, wherein the statin drug and the gliptin drug are administered simultaneously or at different times.

12. A pharmaceutical composition according to claim 1, wherein the composition is provided as a complex agent.

13. A combination preparation for preventing or treating kidney disease or vascular or valvular stenosis, comprising a composition comprising a statin drug and a composition comprising a gliptin drug, wherein each composition is used in combination.

14. A method for preventing or treating vascular or valvular stenosis by administering a statin drug and a gliptin drug in an amount effective for prevention or treatment to a subject in need of prevention or treatment of kidney disease or vascular or valvular stenosis.

15. A method according to claim 14, wherein the statin drug and the gliptin drug are administered simultaneously or at the same time.

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