Pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification, and its preparation method and application

Abstract

The present invention provides a pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification, the pharmaceutical composition containing composition I, composition II and composition III in a mass ratio of (0.1-5):(0.01-0.5):(100-500). The present invention also provides a method for preparing the pharmaceutical composition and its application. The pharmaceutical composition can comprehensively prevent and block cardiovascular calcification, and is particularly suitable for reversing the symptoms of cardiovascular calcification caused by the administration of statin drugs, and also provides the effect of daily supporting and protecting heart health. The pharmaceutical composition of the present invention has each active ingredient stable, high bioavailability, high targeting, no side effects, and can simultaneously achieve the preventive and therapeutic purposes.

Description

[Technical field]

[0001] The present invention relates to the fields of medicine, health food and food technology, and in particular to a pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification and its preparation method and application. [Background technology]

[0002] Vascular calcification is an inappropriate biological mineralization occurring in the soft tissues of the cardiovascular system, a pathological change in which calcium salts are deposited in the arterial wall tissue. It is widely seen in various diseases such as arteriosclerosis, diabetes, nephropathy, senility, and systolic hypertension, and is particularly prone to the development of myocardial infarction and stroke. Previous studies have considered vascular calcification to be a passive, degenerative, and inevitable end-stage process, but recent findings from clinical and basic research suggest that vascular calcification is an active, modifiable, treatable, and preventable process similar to the processes of bone development and cartilage formation.

[0003] Coronary heart disease is the major trigger threatening the health and life of middle-aged and elderly people in modern society. With the progress of society and the improvement of civilization, the prevalence of coronary heart disease increases year by year along with the trend of rejuvenation. Currently, nearly 200 million people worldwide use statins to prevent and treat coronary heart disease, but the latest research has found that people who use statins frequently are more likely to show accelerated coronary artery calcification than those who use them infrequently.

[0004] According to existing pharmacological evidence and clinical trial results, the main physiological mechanism by which statins promote arterial calcification is, first, by inhibiting the function of vitamin K, which leads to arterial calcium deposition; second, by inhibiting the function of coenzyme Q in the body. 10 Inhibits the synthesis of the antioxidant Coenzyme Q 10 The main cause of this is a rapid decline in the antioxidant capacity of the body, which in turn reduces the levels of mitochondrial enzymes and impairs mitochondrial energy metabolism, resulting in a rapid decline in the antioxidant capacity of the body and inducing myocardial damage.

[0005] Therefore, comprehensive prevention and inhibition of cardiovascular calcification is an important key to ensuring the health, reducing side effects, and improving the quality of life of statin users.

[0006] Currently, vitamin K2 is generally used as a dietary supplement to promote calcium absorption, protect joint health, and strengthen bones, and its reverse mineralization effect has not been emphasized or is very little. In addition, vitamin K2 is a fat-soluble vitamin, so it has low solubility in water, is difficult to absorb in the intestine after oral administration, and has low bioavailability. In addition, vitamin K2 is very sensitive to light and heat, and the shelf life of ordinary related products is unstable, and the bioactivity decreases seriously after long-term storage. Therefore, there is an urgent need for vitamin K2-related products with stable activity, high bioavailability, and functional targets. Summary of the Invention

[0007] The objective of the present invention is to provide a Vitamin K2 related product with stable activity, high bioavailability and functional targeting, so as to overcome the above-mentioned shortcomings in the prior art.

[0008] To achieve the above objectives, the present invention provides the following technical solutions: In a first aspect, the present invention provides a pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification, the pharmaceutical composition comprising composition I, composition II and composition III, wherein composition I comprises an active functional ingredient for promoting calcium absorption, composition II comprises an active functional ingredient for inhibiting or reversing calcification, and composition III comprises an active functional ingredient for daily protection of cardiovascular health; The mass ratio of the composition I, the composition II and the composition III is (0.1-5):(0.01-0.5):(100-500).

[0009] In some embodiments, Composition I comprises an active ingredient that promotes calcium absorption, the active ingredient consisting of a main active ingredient and a supplementary active ingredient, the main active ingredient being vitamin D3 and the supplementary active ingredient being one or more of lactose, vitamin A, albumin peptides, vitellogenin peptides, oligofructose, oligogalactose and β-carotene. In yet another embodiment, the composition II comprises an active ingredient for inhibiting or reversing calcification, the active ingredient consisting of a main active ingredient and a supplementary active ingredient, the main active ingredient is vitamin K2, and the supplementary active ingredient is one or more of naringenin, arachidonic acid, tripterin, puerarin, apigenin, and aconite polysaccharide.

[0010] In yet a further embodiment, the composition III comprises an active ingredient for providing daily cardiovascular health protection, the active ingredient being comprised of a primary active ingredient and a secondary active ingredient, the primary active ingredient being Omega-3 fatty acid, pyrroloquinoline quinone, coenzyme Q. 10 and the auxiliary active ingredient is one or more of epicatechin, pumpkin seed oil, phospholipids, krill oil, spirulina, quercetin, lycopene, pomegranate polyphenols, resveratrol, cyanidin, soy isoflavones, and allicin.

[0011] Preferably, the composition I accounts for 0.1% to 1% of the total mass of the drug composition, the composition II accounts for 0.001% to 0.1% of the total mass of the drug composition, and the composition III accounts for 30% to 85% of the total mass of the drug composition.

[0012] Also preferably, the pharmaceutical composition further comprises an auxiliary material portion, and the auxiliary material is one or more of microcrystalline cellulose, silicon dioxide, magnesium stearate, α-cyclodextrin, mannitol, sodium carboxymethylcellulose, hydroxypropylcellulose, glycerol, gelatin, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, talc, modified starch, antioxidants, titanium dioxide, tartrazine aluminum lake, sunset yellow aluminum lake, carmine aluminum coloring agent, brown iron oxide, triacetin, polyethylene glycol, polyvinylpolypyrrolidone, and croscarmellose sodium.

[0013] More preferably, the pharmaceutical composition comprises 0.01-1 part vitamin K2, 300-3000 parts omega-3 fatty acid, 1-40 parts pyrroloquinoline quinone, 10-500 parts coenzyme Q. 10 and 1 to 60 parts by mass of vitamin D3 as a main ingredient.

[0014] In a more preferred embodiment, the vitamin K2 is in the form of MK-7 having an all-trans structure, and the ratio of EPA to DHA in the omega-3 fatty acids is (0.1-4):1. In the second aspect, the present invention also provides the application of said pharmaceutical composition in dietary supplements and health care products.

[0015] In a third aspect, the present invention provides a soft capsule, which contains any of the above-mentioned pharmaceutical compositions containing vitamin K2 for improving cardiovascular calcification.

[0016] In a preferred embodiment of the soft gelatin capsule, the soft gelatin capsule contains 0.01 to 0.7 parts of vitamin K2, 500 to 2500 parts of omega-3 fatty acid, 1 to 30 parts of pyrroloquinoline quinone, 50 to 500 parts of coenzyme Q 10The ingredients include 1 to 50 parts by weight of vitamin D3, 50 to 500 parts by weight of soft capsule shells, 10 to 100 parts by weight of enteric coating powder, and 20 to 200 parts by weight of auxiliary materials.

[0017] In a fourth aspect, the present invention also provides a method for preparing the above soft capsule, the method comprising the steps of: (1) Granulation: Under light-shielding conditions, the fat-insoluble components of the composition II and composition III described in claim 1 and some auxiliary materials are uniformly mixed, and granulated by a dry granulation method in which the particle size is controlled to 50 to 200 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into tablets of a predetermined weight using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a normal coating procedure. (4) Capsule liquid: The fat-soluble component in composition III, composition I, and the antioxidant in the auxiliary materials are weighed, and the antioxidant and composition I are each dissolved in the oil of the fat-soluble component in composition III at a temperature of 20 to 60°C to obtain a mixed oil, which is then left to stand for 4 to 12 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell is obtained by adopting the general recipe and preparation method in the industry; (6) Soft capsule: The encapsulation liquid of step (4) is injected between the two capsule shell layers containing the coated tablet of step (3), and the tablet is pressed to obtain a soft capsule containing the tablet. Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] (1) It can target and block and prevent calcification of the cardiovascular system. On the one hand, it can promote calcium absorption and avoid the accumulation of excess calcium in the vascular wall; on the other hand, it can enhance the conversion of incorrectly deposited calcium; on the third side, it can improve myocardial capacity, strengthen resistance, and enhance mitochondrial energy metabolism, thereby avoiding damage to the organism, and from a macroscopic perspective, it can implement pre-prevention, block the cause of risk, and provide a pre-planned protective measure. In addition, it can improve vascular calcification and at the same time prevent and eliminate other risks caused by vascular calcification. (2) The products obtained using this preparation method have a special double protection method that avoids the influence of the environment such as light, heat, and acid, and the first-pass effect in the oral cavity and gastrointestinal tract, making the activity of MK-7 and other functional ingredients in the composition more stable and long-lasting, with higher bioavailability, more stable product quality, and a longer shelf life. (3) In addition to improving calcification, it also has the effects of improving the body's immune system, acting as an antioxidant, anti-cancer, preventing aging, and strengthening bones. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 shows the trend of the effect of drug compositions on OPG (osteoprotegerin) levels in rat serum. [Diagram 2] FIG. 1 shows the trend of the effect of drug compositions on OPN (osteopontin) levels in rat serum. [Diagram 3] FIG. 1 shows the trend of the effect of drug compositions on the level of CaBP (calcium binding protein) in rat serum. [Figure 4] FIG. 1 shows the trend of the effect of drug compositions on the levels of BMP-7 (bone morphogenetic protein-7) in rat serum. [Diagram 5] FIG. 1 shows the trend of the effect of drug compositions on calcium levels in the arterial blood vessels of rats. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The technical solutions in the embodiments of the present invention will be described below clearly and completely with reference to the embodiments of the present invention and the accompanying drawings, but it is clear that the described embodiments are only some of the embodiments of the present invention, and are not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts fall within the protection scope of the present invention.

[0021] The present invention provides a pharmaceutical composition containing vitamin K2 (MK-7) for improving cardiovascular calcification and its preparation method, said pharmaceutical composition can comprehensively prevent and block cardiovascular calcification, and is particularly suitable for reversing the symptoms of cardiovascular calcification caused by the administration of statin drugs, while providing a beneficial daily support and protection effect for the heart.

[0022] The present invention improves cardiovascular calcification from three perspectives: promoting calcium absorption, inhibiting calcification, and daily protection. On the one hand, it promotes calcium absorption to avoid inappropriate calcium accumulation in the vascular wall; on the other hand, it strengthens the promotion of the degeneration of incorrectly deposited calcium; and thirdly, it strengthens mitochondrial energy metabolism to improve myocardial capacity, strengthen resistance, and avoid damage to the body, thereby achieving the effect of preventing risks in advance from a macroscopic perspective, blocking the causes of risks, and providing pre-planned protective measures. The synergistic action of these three perspectives achieves the comprehensive effect of improving cardiovascular calcification.

[0023] In one aspect, the present invention provides a pharmaceutical composition containing vitamin K2 that improves cardiovascular calcification, the pharmaceutical composition comprising composition I, composition II, and composition III; The mass ratio of the composition I, the composition II, and the composition III is (0.1-5):(0.01-0.5):(100-500).

[0024] The composition I contains an active ingredient that promotes calcium absorption, and may contain one or more of lactose, vitamin A, vitamin D3, albumin peptide, vitellogenin peptide, oligofructose, oligogalactose, and β-carotene.

[0025] Vitamin D3, also known as cholecalciferol, is a type of vitamin D that improves calcium and phosphorus absorption in the body and saturates plasma calcium and plasma phosphorus concentrations. Vitamin D3 is a precursor of D hormone, and research has revealed that it is hydroxylated in the kidneys and lungs to become active vitamin D, which acts in the kidneys and intestines to promote calcium reabsorption. Therefore, the present invention preferably uses VD3 and MK-7 in combination, which on the one hand promotes calcium absorption and avoids excessive deposition of absorbed calcium, and on the other hand plays the role of MK-7 to activate osteocalcin, which acts as a calcium claw to capture deposited calcium in bones, thereby inhibiting and preventing calcification.

[0026] Preferably, vitamin D3 is used as the main component of composition I of the present invention.

[0027] More preferably, the vitamin D3 in the present invention is derived from a plant and has an effective activity unit of 400 IU or more.

[0028] The composition II comprises an active ingredient capable of inhibiting or reversing calcification, and may contain one or more of naringenin, arachidonic acid, tripterin, puerarin, apigenin, aconite polysaccharide, and vitamin K2.

[0029] Vitamin K2 is a type of fat-soluble vitamin, mainly used for the treatment and prevention of osteoporosis. According to the length of the tail chain, K2 is subdivided into subgroups such as MK-4, MK-7, MK-8, and MK-10. All vitamin K2 are structurally similar, but differ in the length of the side chain. The longer the side chain, the better it is absorbed, the more biologically active it is, and the longer it remains in the blood. Therefore, the long-chain MK-7 has been shown to be of the best quality, with the highest bioavailability after oral intake and a long half-life, and as a result, it can exert its effects over a longer time cycle. Also, MK-7 is obtained by plant fermentation, making it a healthier source.

[0030] Osteocalcin, also known as bone gamma-carboxyglutamic acid protein, is a protein secreted by osteoblasts. It is a vitamin K-dependent protein and a specific executor of calcium into bone. When secreted, it has no physiological activity and needs to undergo carboxylation to play a physiological role. After carboxylation, it can firmly grasp calcium salts, promote the deposition of calcium salts in bones, and increase the rate of bone mineralization. Therefore, vitamin K2 is considered to be crucial for maintaining bone health and preventing osteoporosis. When there is a lack of vitamin K2 in the body, osteocalcin cannot form calcium nails, just like a human hand without a thumb, and the grip strength is reduced and a large amount of calcium is lost from the bones, leading to osteoporosis.

[0031] In addition to being involved in normal bodily functions and metabolism, many calcium ions in the blood have two different destinations: (1) deposition in appropriate locations such as bones and teeth, and (2) deposition in incorrect locations such as articular cartilage, arteries and blood vessels in the heart and brain, and soft tissues such as the gallbladder and kidneys. Calcium deposited in the incorrect locations can cause dysfunction of tissues and organs in those locations. If most of the calcium in the blood does not migrate to bones, it is deposited in large amounts in soft tissues such as blood vessels. Long-term accumulation causes calcification of organs, especially vascular calcification. However, the body has mechanisms to prevent this abnormal deposition, and many studies have shown that vitamin K2 has the ability to prevent this incorrect deposition of calcium. Carboxylated matrix gamma-carboxyglutamic acid protein can prevent calcium deposition in soft tissues such as blood vessels and cartilage, bind to calcium ions deposited in the vascular wall, and move them out of the blood vessels, thereby improving arterial calcification.

[0032] Therefore, the present invention preferably utilizes as its core difference the ability of vitamin K2 to activate osteocalcin and precisely capture erroneously deposited calcium to achieve the goal of targeted blockade and prevention of cardiovascular calcification.

[0033] More preferably, the vitamin K2 is of MK-7 structure and is obtained by organic bean fermentation method, which, compared with chemical synthesis method, has no organic reagent residue, the whole process is pollution-free, and is safer to consume.

[0034] More preferably, the MK-7 is in the all-trans configuration and has a purity of 99.5% or more, and the bioavailability of the MK-7 in the trans configuration is higher.

[0035] Composition III contains active ingredients that provide daily cardiovascular health protection, including omega-3 fatty acids, pyrroloquinoline quinone (PQQ), coenzyme Q, and 10 (CoQ 10), epicatechin, pumpkin seed oil, phospholipids, krill oil, spirulina, quercetin, lycopene, pomegranate polyphenols, resveratrol, cyanidin, soy isoflavones and allicin.

[0036] Coenzyme Q 10 is a fat-soluble quinone ring compound with a structure similar to vitamin K, and was named after the degree of polymerization of the side chain-polyisopentenyl group at the 6th position of the parent nucleus, which is 10. Statins, which lower cholesterol, reduce the amount of coenzyme Q in the body. 10 Some studies have shown that statins, along with coenzyme Q, can lower levels of the hormone by as much as 40%, worsening heart problems. 10 Taking coenzyme Q can help reduce this side effect of statins, while also relieving drug-induced muscle pain and fatigue and acting as a liver protector. 10 has the functions of activating somatic cells, generating energy, improving immunity, enhancing antioxidant power, delaying aging, and increasing the vitality of the body.

[0037] PQQ is produced by gram-negative bacteria and has a wide range of nutritional effects on microorganisms, animals and plants, and has antioxidant properties. The protective effect of PQQ on the heart is related to its ability to scavenge free radicals. PQQ can scavenge the reactive oxygen species generated by hypoxia-reperfusion, significantly reducing the release of lactate dehydrogenase in the heart, and through the catalysis of the enzyme flavin reductase, the catalytic product of PQQ also reduces the peroxidation state of hemoglobin and eliminates myocardial damage caused by hypoxia-reperfusion. PQQ reduces the peroxidation state of hemoglobin and eliminates myocardial damage caused by hypoxia-reperfusion. Research has shown that using PQQ to protect the hearts of ischemia-reperfusion mice can significantly reduce the extent of myocardial infarction, enhance the rise and fall rate of left ventricular pressure and left ventricular diastolic pressure, reduce ventricular fibrillation, and reduce the level of malondialdehyde in myocardial tissue. PQQ also inhibited the production of ROS and the decline in mitochondrial membrane potential in rat cardiomyocytes induced by hydrogen peroxide, thereby reducing oxidative stress, inhibiting the inactivation of mitochondrial function, and protecting rat cardiomyocytes.

[0038] Therefore, in the present invention, the amount of coenzyme Q in the body caused by long-term administration of statin drugs is 10 In light of the decline in levels, PQQ and CoQ 10 It is preferable to incorporate coenzyme Q into the body. 10 On the other hand, it activates cells, enhances mitochondrial energy metabolism, and improves the myocardial ability to avoid drug damage. 10 Both can improve the body's antioxidant capacity and maintain the balance of scavenging free radicals.

[0039] Preferably, the PQQ is in the form of the disodium salt of pyrroloquinoline quinone or the acid form of pyrroloquinoline quinone that is 99% or greater pure.

[0040] Preferably, CoQ 10 has a purity of 98% or more.

[0041] Omega-3 is a long-chain polyunsaturated fatty acid found in plants and marine organisms. Existing research data shows that Omega-3 has the effect of promoting heart health and preventing coronary artery disease. Omega-3 is effective in preventing and treating cardiovascular and cerebrovascular diseases mainly by protecting the cells lining blood vessels, restoring vascular elasticity, relaxing blood vessels, and inhibiting platelet aggregation, lowering blood pressure, inhibiting the formation of blood clots, while reducing blood lipids.

[0042] In addition, Omega-3 also plays a role in preventing arrhythmia and sudden cardiac death. A clinical study that followed more than 11,000 patients with coronary artery disease revealed that taking 1 to 2 grams of Omega-3 unsaturated fatty acids per day significantly reduced the mortality rate of cardiovascular disease patients, and in particular reduced the incidence of sudden cardiac death by 45%.

[0043] There are three types of omega-3 fatty acids: alpha-linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). EPA has the effect of preventing the onset of stroke and myocardial infarction, lowering blood cholesterol, and preventing arteriosclerosis. DHA activates brain cells, promotes the adjustment of neural circuits, and maintains the normal function of brain cells. Supplementing with DHA can improve concentration, learning disabilities, memory loss, and Alzheimer's disease.

[0044] Omega-3 also has the effects of resisting nervous system diseases, preventing and resisting cancer, anti-inflammatory, etc. Therefore, the use of Omega-3 supplementation in the pharmaceutical composition of the present invention as part of a lifestyle intervention reduces the risk of mortality and sudden cardiac death in the population.

[0045] Preferably, in the present invention, the ratio of EPA:DHA in Omega-3 fatty acids is (0.1-4:1).

[0046] More preferably, the content of EPA in the Omega-3 fatty acids is ≧40%.

[0047] More preferably, the content of EPA in the Omega-3 fatty acids is ≧45%.

[0048] The present invention finds that there are many active ingredients or dietary supplements in the field that can comprehensively block and prevent cardiovascular calcification, but they are not simply combined to achieve synergistic effects.Therefore, the present invention starts from the mechanism of calcification and cardiovascular disease and the metabolic characteristics of statin drugs, and through many tests, screens the above ingredients and uses them in combination, which on the one hand promotes calcium absorption to avoid excess calcium accumulation in vascular walls, on the other hand strengthens the conversion of calcium that is mistakenly deposited, and on the third side, improves myocardial capacity through strengthening mitochondrial energy metabolism, and enhances the resistance of the body to prevent damage, so as to macroscopically implement pre-prevention, block danger sources, and provide pre-planned protection measures.While improving vascular calcification, it can also prevent and eliminate other risks caused by vascular calcification.

[0049] In one specific embodiment of the present invention, a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification contains the following components by mass: 0.01 to 1 part vitamin K2 (MK-7), 300 to 3,000 parts omega-3 fatty acid, 1 to 40 parts pyrroloquinoline quinone (PQQ), and 10 to 500 parts coenzyme Q. 10 (CoQ 10 ) and 1 to 60 parts vitamin D3 (VD3).

[0050] In a more preferred embodiment, the composition further comprises auxiliary ingredients, which are auxiliary ingredients necessary for the preparation of the pharmaceutical composition containing vitamin K2 (MK-7) for improving cardiovascular calcification provided by the present invention, including coating powder, antioxidant, lubricant, and capsule shell. More preferably, the auxiliary ingredients are one or more of microcrystalline cellulose, silicon dioxide, magnesium stearate, α-cyclodextrin, mannitol, sodium carboxymethylcellulose, hydroxypropylcellulose, glycerol, gelatin, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, talc, modified starch, antioxidant, titanium dioxide, tartrazine aluminum lake, sunset yellow aluminum lake, carmine aluminum coloring agent, brown iron oxide, triacetin, polyethylene glycol, polyvinylpolypyrrolidone, and croscarmellose sodium.

[0051] The pharmaceutical composition may be in the form of one or more selected from the group consisting of tablets, powders, hard capsules, soft capsules, and suspensions.

[0052] The second aspect of the present invention provides the application of the above pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification in dietary supplements and health care products.

[0053] In a third aspect, the present invention provides a soft capsule, which contains the above pharmaceutical composition containing vitamin K2 for improving cardiovascular calcification. Preferably, the soft capsule comprises the following components by weight: 0.01-0.7 parts of vitamin K2 (MK-7), 500-2500 parts of omega-3 fatty acid, 1-30 parts of pyrroloquinoline quinone (PQQ), and 50-500 parts of coenzyme Q. 10 (CoQ 10 ), 1-50 parts Vitamin D3 (VD3), 50-500 parts soft capsule shells, 10-200 parts enteric coating powder, 100 parts, and 20-200 parts auxiliary materials.

[0054] In a fourth aspect, the present invention provides a method for preparing the pharmaceutical composition containing vitamin K2 (MK-7) for improving cardiovascular calcification, the method comprising the steps of: (1) Granulation: Under light-shielding conditions, the fat-insoluble components of Composition II and Composition III and some of the auxiliary materials are uniformly mixed, and granulated by a dry granulation method in which the particle size is controlled to 50 to 200 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into tablets of a predetermined weight using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a normal coating procedure. (4) Capsule liquid: The fat-soluble component in composition III, composition I, and the antioxidant in the auxiliary materials are weighed, and the antioxidant and composition I are each dissolved in the oil of the fat-soluble component in composition III at a temperature of 20 to 60°C to obtain a mixed oil, which is then left to stand for 4 to 12 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell is obtained by adopting the general recipe and preparation method in the industry; (6) Soft capsule: The encapsulation liquid of step (4) is injected between the two capsule shell layers containing the coated tablet of step (3), and the tablet is pressed to obtain a soft capsule containing the tablet.

[0055] The preparation method of the present invention may also include further processing methods including, but not limited to, drying, tablet washing, tablet extraction, packaging, etc., to ultimately obtain the finished product.

[0056] The product obtained using the preparation method of the present invention has a special double protection method that avoids light, heat, acid and other environmental influences, as well as the first-pass effect in the oral cavity and stomach, and prevents premature decomposition after ingestion, making the activity of MK-7 and other active ingredients in the composition more stable and long-lasting, with higher bioavailability, more stable product quality, and longer shelf life.

[0057] The third aspect of the present invention provides a dietary supplement and the application of the above pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification.

[0058] The following examples are used to illustrate the present invention, but are not intended to limit the protection scope of the present invention.

[0059] Example 1 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, which contains the following ingredients by weight: 0.18 parts Vitamin K2 (MK-7), 1200 parts Omega-3 fatty acids, 10 parts PQQ, 200 parts Coenzyme Q 10 (CoQ 10 ), 5 parts Vitamin D3 (VD3), 250 parts soft capsule shells, 100 parts enteric coating powder and 50 parts auxiliary materials.

[0060] The preparation method is as follows. (1) Granulation: In the dark, mix equal amounts of MK-7, PQQ and tablet auxiliary materials, adjust the particle size to 100 mesh, and perform dry granulation to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into plain tablets of a predetermined weight of 0.2 g using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a conventional coating process. (4) Capsule liquid: Omega-3, CoQ 10 , VD3 and antioxidants were weighed out according to weight, and antioxidants and CoQ were added to Omega-3 oil. 10 , and VD3 are dissolved at a temperature of 40°C to obtain a mixed oil solution, which is then left to stand for 12 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The preparation of the soft capsule shell is obtained by adopting the general recipe and preparation method in this industry. (6) Soft capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shells containing the coated tablets from (3) and the tablets are pressed through a special die to obtain a soft capsule containing the tablets inside.

[0061] Example 2 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, the pharmaceutical composition comprising the following components by weight: 0.09 parts Vitamin K2 (MK-7), 600 parts Omega-3 fatty acids, 10 parts PQQ, 100 parts Coenzyme Q 10 (CoQ 10 ), 2 parts Vitamin D3 (VD3), 150 parts soft capsule shells, 80 parts enteric coating powder and 30 parts auxiliary materials.

[0062] The preparation method is as follows. (1) Granulation: Under light-shielded conditions, mix equal amounts of MK-7, PQQ and tablet auxiliary materials, and use dry granulation to obtain granules with a particle size of 120 mesh. (2) Tablets: The granules obtained in step (1) are processed into uncoated tablets of the specified weight of 0.15 g using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a conventional coating process. (4) Capsule liquid: Omega-3, CoQ10 , VD3 and antioxidants were weighed out according to weight, and antioxidants and CoQ were added to Omega-3 oil. 10 , and VD3 are dissolved at a temperature of 40°C to obtain a mixed oil solution, which is then left to stand for 12 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The preparation of the soft capsule shell is obtained by adopting the general recipe and preparation method in this industry. (6) Soft capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shell layers containing the coated tablets from step (3) and the tablets are pressed through a special die to obtain a soft capsule containing the tablets inside.

[0063] Example 3 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, the pharmaceutical composition containing the following components in the weight ratio: 0.12 parts Vitamin K2 (MK-7), 900 parts Omega-3 fatty acids, 15 parts PQQ, 150 parts Coenzyme Q 10 (CoQ 10 ), 3 parts Vitamin D3 (VD3), 180 parts soft capsule shells, 70 parts enteric coating powder and 40 parts auxiliary materials.

[0064] The preparation method is as follows. (1) Granulation: Under light-shielded conditions, mix equal amounts of MK-7, PQQ and tablet auxiliary materials, and use dry granulation to control the particle size to 80 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into plain tablets of a predetermined weight of 0.25 g using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a conventional coating process. (4) Capsule liquid: Omega-3, CoQ 10 The antioxidants, VD3 and CoQ were weighed out and added to Omega-3 oil at 50°C. 10 , VD3 is dissolved to obtain an oil-liquid mixture, which is then left to stand for 12 hours to degas and obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell can be prepared by adopting the general recipe and preparation method in this industry. (6) Soft capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shells containing the coated tablets from step (3) and the tablets are pressed through a special die to obtain a soft capsule containing the tablets inside.

[0065] Example 4 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, which contains the following components by weight: 0.45 parts Vitamin K2 (MK-7), 1000 parts Omega-3 Fatty Acids, 12 parts PQQ, 250 parts Coenzyme Q 10 (CoQ 10 ), 4 parts Vitamin D3 (VD3), 180 parts soft capsule shells, 90 parts enteric coating powder and 45 parts auxiliary materials.

[0066] The preparation method is as follows. (1) Granulation: Under light-shielded conditions, mix equal amounts of MK-7, PQQ and tablet auxiliary materials, and use dry granulation to control the particle size to 100 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into plain tablets of a predetermined weight of 0.26 g using a tablet machine. (3) Coated tablets: The plain tablets prepared in step (2) are mixed with an enteric coating powder in a coating pot to produce coated tablets according to normal coating procedures. (4) Capsule liquid: Omega-3, CoQ 10 , VD3 and antioxidants were weighed out according to weight, and antioxidants and CoQ were added to Omega-3 oil. 10 , and VD3 are dissolved at a temperature of 45°C to obtain a mixed oil solution, which is then left to stand for 8 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell can be prepared by adopting the general recipe and preparation method in this industry. (6) Soft capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shells containing the coated tablets from step (3) and the tablets are pressed through a special die to obtain a soft capsule containing the tablets inside.

[0067] Example 5 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, which contains the following components by weight: 0.65 parts Vitamin K2 (MK-7), 1300 parts Omega-3 fatty acids, 20 parts PQQ, 200 parts Coenzyme Q 10 (CoQ 10 ), 5 parts Vitamin D3 (VD3), 300 parts soft capsule shells, 100 parts enteric coating powder and 50 parts auxiliary materials.

[0068] The preparation method is as follows. (1) Granulation: Under light-shielded conditions, equal amounts of MK-7, PQQ and a tableting aid are mixed, and granulated by dry granulation with the particle size controlled to 120 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into uncoated tablets of the specified weight of 0.3 g using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a conventional coating process. (4) Capsule liquid: Omega-3, CoQ 10 , VD3 and antioxidants were weighed out according to weight, and antioxidants and CoQ were added to Omega-3 oil. 10 , and VD3 are dissolved at a temperature of 40°C to obtain a mixed oil solution, which is then left to stand for 10 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell can be prepared by adopting the general recipe and preparation method in this industry. (6) Soft capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shells containing the coated tablets from step (3) and the tablets are pressed through a special die to obtain a soft capsule containing the tablets inside.

[0069] Example 6 This example provides a pharmaceutical composition containing vitamin K2 (MK-7) that improves cardiovascular calcification, which contains the following components by weight: 0.3 parts Vitamin K2 (MK-7), 1500 parts Omega-3 fatty acids, 18 parts PQQ, 180 parts Coenzyme Q 10 (CoQ 10 ), 10 parts Vitamin D3 (VD3), 200 parts soft capsule shells, 80 parts enteric coating powder and 60 parts auxiliary materials.

[0070] The preparation method is as follows. (1) Granulation: Under light-shielded conditions, mix equal amounts of MK-7, PQQ and tablet auxiliary materials, and use dry granulation to control the particle size to 100 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into plain tablets of a predetermined weight of 0.26 g using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a conventional coating process. (4) Capsule liquid: Omega-3, CoQ 10 , VD3 and antioxidants were weighed out according to weight, and antioxidants and CoQ were added to Omega-3 oil. 10 , and VD3 are dissolved at a temperature of 40°C to obtain a mixed oil solution, which is then left to stand for 11 hours and degassed to obtain a capsule liquid. (5) Soft capsule shell: The soft capsule shell can be prepared by adopting the general recipe and preparation method in this industry. (6) Soft Gelatin Capsule: Using special equipment, the encapsulation liquid from step (4) is injected between the two capsule shells containing the coated tablet from step (3) and the tablet is pressed through a special die to obtain a soft capsule containing the tablet inside.

[0071] Validation Testing 1. Examination of accelerated stability of MK-7 content in the composition The soft capsules produced in Examples 4, 5 and 6 were used as the experimental group, and commercially available vitamin K2 (MK-7) soft capsules (MK-7 content 90 μg) were used as the control group, and were filled into sealed brown vials. The accelerated experiment was carried out in a constant temperature and humidity room at 40 ± 2 ° C and 75% ± 5% RH. To measure the MK-7 content, samples were taken at 0, 1, 2, 3, 4, 5, 6 and 9 months. The MK-7 content was measured by high performance liquid chromatography (HPLC). The results of the accelerated stability test are shown in the table below.

[0072] [Table 1] As can be seen from the results in Table 1, the content of MK-7 in the compositions of Example 4, Example 5 and Example 6 remained essentially unchanged and maintained stability, while that of the control group began to decay after 3 months of accelerated conversion. This indicates that the MK-7 active ingredient in the MK-7-containing drug composition prepared by the preparation method of the present invention can be stably maintained for a longer period of time. Based on the normal accelerated conversion time, a shelf life of at least 3 years can be ensured at room temperature.

[0073] 2. Effect of the pharmaceutical composition on vascular calcification in rats SPF grade SD male rats were selected, healthy, 6–8 weeks old, and weighing 180 g–200 g. The rats were randomly divided into a normal control group (10 rats) and a dexamethasone injection group (100 rats) according to their body weight. The rats in the dexamethasone injection group were intramuscularly injected with 2.5 mg / kg dexamethasone three times a week, and the rats in the normal control group were intramuscularly injected with an equivalent amount of saline. After six weeks of injections, the injections were stopped. The rats were fasted for 24 hours, blood was collected from the orbital vein, serum was separated, and serum alkaline phosphatase (ALP) values ​​were measured using an automatic biochemistry analyzer, and the ALP values ​​of the dexamethasone injection group were compared with those of the normal control group to evaluate the modeling results. The rats that were successfully modeled were randomly divided into five groups: model group, positive control group (pure MK-7 100 μg / kg), and experimental groups (Example 4, Example 5, Example 6). The normal control group and model group were orally administered 10mg / kg of edible oil, the experimental group was orally administered the corresponding MK-7 according to the established practice, and the positive control group was orally administered MK-7 (12mg / kg) dissolved in edible oil once a day for 6 weeks. The rats in each group were kept in separate cages with free access to food and water, and the room temperature was kept constant at (22±2)℃.

[0074] Observation index and method: After the final administration, the rats were fasted for 24 hours, blood was collected from the orbital vein, serum was separated, and serum OPG, OPN, CaBP, and BMP-7 concentrations were measured by radioimmunoassay.

[0075] Measurement of vascular calcium content: 10 mg of abdominal aorta was dissolved in nitric acid, digested, calcined, and then redissolved in deionized water containing 27 nmol / L KCL and 27 μmol / L LaCl3. The absorbance at 442.7 nm was read using an atomic absorption spectrophotometer and converted to tissue calcium content (μmol / mg.dw).

[0076] SPSS21.0 software was used for statistical analysis, and the measured data were expressed as mean ± standard deviation and analyzed by one-way analysis of variance and Dunnett-t test, with P < 0.05 considered as a statistically significant difference.

[0077] Combining Figures 1 to 5, the effect of the drug combination on vascular calcification in rats can be seen. Compared with the normal control group, the serum OPN (osteopontin), OPG (osteoprotegerin), CaBP (calcium binding protein) and BMP-7 (bone morphogenetic protein-7) levels of rats in the model group were significantly decreased. Compared with the model group, the serum OPN (osteopontin), OPG (osteoprotegerin), CaBP (calcium binding protein) and BMP-7 (bone morphogenetic protein-7) levels of rats in the experimental group (Example 4), experimental group (Example 5) and experimental group (Example 6) were significantly increased, and the difference is obvious. Meanwhile, compared with the normal control group, the vascular calcium level of rats in the model group was significantly increased, and compared with the model group, the vascular calcium level of rats in the experimental group (Example 4), experimental group (Example 5) and experimental group (Example 6) was significantly decreased, and the difference is obvious.

[0078] On the other hand, OPN (osteopontin) is a marker gene for the conversion of vascular smooth muscle from a contractile phenotype to a synthetic phenotype, and can suppress calcification of cultured vascular smooth muscle cells. OPG (osteoprotegerin) can be expressed and released in vascular endothelial cells and smooth muscle cells, and prevents the vascular damaging effects of inflammatory cytokines by enhancing the survival of endothelial cells, and is associated with the rupture of atherosclerotic plaques. BMP-7 (bone morphogenetic protein-7) is a bone morphogenetic protein that belongs to the transforming growth factor β family and can improve vascular calcification by suppressing the proliferation of vascular smooth muscle cells VSMC. Calcium-binding protein (CaBP) is involved in various vital phenomena, such as the regulation of the cell cycle, cell proliferation and differentiation, angiogenesis, and the reconstruction of the extracellular matrix, and has the proliferation-promoting and survival-promoting effects of cardiomyocytes. It promotes calcium and phosphorus absorption, as well as the release and transport capacity of bone calcium.

[0079] In summary, the pharmaceutical composition of the present invention can regulate the inhibition of vascular calcification by significantly increasing the levels of OPN (osteopontin), OPG (osteoprotegerin), CaBP (calcium binding protein) and BMP-7 (bone morphogenetic protein-7), thus reducing vascular calcium content. Also, through the experimental group and the positive control group, it was found that the pharmaceutical composition of the present invention has a combined effect and a synergistic effect compared with a single compound.

[0080] The above is merely a preferred embodiment of the present invention, and is not a formal limitation of the present invention. Although the present invention is disclosed in the preferred embodiment as above, it is not intended to limit the present invention, and a person skilled in the art can use the above disclosed method and its technical content to modify the above into an equivalent embodiment with similar changes without departing from the scope of the technical solution of the present invention. However, simple changes, equivalent changes and modifications made to the above embodiment according to the technical essence of the present invention without departing from the content of the technical concept of the present invention are still within the scope of the technical concept of the present invention.

Claims

1. A pharmaceutical composition comprising Composition I, Composition II, and Composition III, wherein Composition I comprises an active functional ingredient that promotes calcium absorption, Composition II comprises an active functional ingredient that inhibits or reverses calcification, and Composition III comprises an active functional ingredient that routinely protects cardiovascular health; A vitamin K composition for improving cardiovascular calcification, wherein the mass ratio of the composition I, the composition II and the composition III is (0.1-5):(0.01-0.5):(100-500). 2 A pharmaceutical composition comprising:

2. The composition I contains an active ingredient that promotes calcium absorption, the active ingredient being composed of a main active ingredient and a supplementary active ingredient, and the main active ingredient is vitamin D 3 and the auxiliary active ingredient is one or more of lactose, vitamin A, albumin peptide, vitellogenin peptide, oligofructose, oligogalactose, and β-carotene. 2 A pharmaceutical composition comprising:

3. The composition II comprises an active ingredient that inhibits or reverses calcification, the active ingredient being composed of a main active ingredient and a supplementary active ingredient, the main active ingredient being vitamin K 2 2. The vitamin K for improving cardiovascular calcification according to claim 1, wherein the auxiliary active ingredient is one or more of naringenin, arachidonic acid, tripterin, puerarin, apigenin, and aconite polysaccharide. 2 A pharmaceutical composition comprising:

4. Composition III comprises an active ingredient for providing daily cardiovascular health protection, the active ingredient being comprised of a main active ingredient and a supplementary active ingredient, the main active ingredient being Omega-3 fatty acid, pyrroloquinoline quinone, coenzyme Q 10 and the auxiliary active ingredient is one or more of epicatechin, pumpkin seed oil, phospholipid, krill oil, spirulina, quercetin, lycopene, pomegranate polyphenol, resveratrol, cyanidin, soybean isoflavone, and allicin. 2 A pharmaceutical composition comprising:

5. The vitamin K composition for improving cardiovascular calcification according to any one of claims 1 to 4, characterized in that the composition I accounts for 0.1% to 1% of the total mass of the pharmaceutical composition, the composition II accounts for 0.001% to 0.1% of the total mass of the pharmaceutical composition, and the composition III accounts for 30% to 85% of the total mass of the pharmaceutical composition. 2 A pharmaceutical composition comprising:

6. The vitamin K for improving cardiovascular calcification according to any one of claims 1 to 4, characterized in that the pharmaceutical composition further comprises an auxiliary material portion, and the auxiliary material is one or more of microcrystalline cellulose, silicon dioxide, magnesium stearate, α-cyclodextrin, mannitol, sodium carboxymethylcellulose, hydroxypropylcellulose, glycerol, gelatin, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, talc, modified starch, antioxidant, titanium dioxide, tartrazine aluminum lake, sunset yellow aluminum lake, carmine aluminum coloring agent, brown iron oxide, triacetin, polyethylene glycol, polyvinylpolypyrrolidone, and croscarmellose sodium. 2 A pharmaceutical composition comprising:

7. The pharmaceutical composition contains 0.01 to 1 part of vitamin K 2 , 300-3000 parts Omega-3 fatty acids, 1-40 parts pyrroloquinoline quinone, 10-500 parts coenzyme Q 10 and 1 to 60 parts of vitamin D 3 The vitamin K composition for improving cardiovascular calcification according to any one of claims 1 to 4, characterized in that it contains a main component of 2 A pharmaceutical composition comprising:

8. Vitamin K 2 is a form of MK-7 with an all-trans structure, Vitamin K for improving cardiovascular calcification according to any one of claims 1 to 4, characterized in that the ratio of EPA to DHA in the Omega-3 fatty acids is (0.1-4):

1. 2 A pharmaceutical composition comprising:

9. The vitamin K composition according to any one of claims 1 to 8 that improves cardiovascular calcification 2 The pharmaceutical composition comprising the compound is applied in dietary supplements and health care products.

10. The vitamin K composition according to any one of claims 1 to 8 that improves cardiovascular calcification 2 A soft capsule comprising a drug composition comprising:

11. 0.01 to 0.7 parts vitamin K 2 , 500-2500 parts Omega-3 fatty acids, 1-30 parts pyrroloquinoline quinone, 50-500 parts coenzyme Q 10 , 1 to 50 parts vitamin D 3 The soft capsule according to claim 9, characterized in that the soft capsule is composed of the following components: 50 to 500 parts of the soft capsule shell, 10 to 100 parts of the enteric coating powder, and 20 to 200 parts of the auxiliary material.

12. (1) Granulation: Under light-shielding conditions, the fat-insoluble components of the composition II and composition III according to claim 1 and some of the auxiliary materials are uniformly mixed, and granulated by a dry granulation method in which the particle size is controlled to 50 to 200 mesh to obtain granules. (2) Tablets: The granules obtained in step (1) are processed into tablets of a predetermined weight using a tablet machine. (3) Coated tablets: The plain tablets obtained in step (2) are mixed with an enteric coating powder in a coating pot, and coated tablets are produced according to a normal coating procedure. (4) Capsule liquid: The fat-soluble component in the composition III described in claim 1, the composition I described in claim 1, and the antioxidant in the auxiliary material described in claim 6 are weighed, and the antioxidant and the composition I are dissolved in the oil of the fat-soluble component in the composition III at a temperature of 20 to 60°C to obtain a mixed oil, which is left for 4 to 12 hours to be degassed, to obtain a capsule liquid; (5) Soft capsule shell: The soft capsule shell is obtained by using the general recipe and preparation method in the industry; (6) Soft capsule: The method for preparing a soft gelatin capsule according to claim 10 or 11, further comprising the steps of injecting the capsule liquid of step (4) between the two capsule shell layers containing the coated tablet of step (3) and pressing the tablet to obtain a soft capsule containing the tablet.

Images