A pharmaceutical composition containing MK-7 and its application and preparation method

Abstract

The present invention belongs to the field of medical technology, and relates to a pharmaceutical composition containing MK-7, and its application and preparation method. The pharmaceutical composition comprises MK-7, and at least one pharmaceutically acceptable excipient; the MK-7 is contained in liposome powder; the preparation of liposome powder comprises the following steps: (1) MK-7, an emulsifier and water are mixed and emulsified to obtain a premixed liquid to obtain an emulsion; (2) the emulsion is mixed with the excipient to obtain liposome powder; in step (1), the ratio of MK-7, an emulsifier and water is (0.2-10): (0.05-3): (0.1-8). The present invention prepares MK-7 raw material into liposome powder, which can greatly improve the stability of the active pharmaceutical ingredient MK-7 in the preparation, and the pharmaceutical composition can improve bone density, promote osteocalcin carboxylation by increasing the levels of osteocalcin and alkaline phosphatase (ALP), and treat osteoporosis, which has a wide range of application prospects.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a pharmaceutical composition containing MK-7 and an application and a preparation method thereof. Background Art

[0002] Vitamin K is a type of fat-soluble compound with a quinone structure. There are two main types of vitamin K in nature. The first is vitamin K1, also known as phylloquinone, which mainly comes from plants; the second is vitamin K2. Vitamin K has two forms, MK-4 and MK-7. The former is vitamin K2 with 4 isoprene units on the side chain, and the latter is vitamin K2 with 7 isoprene units on the side chain. Vitamin K2 is a fat-soluble vitamin that has biological activity in animals and is one of the important vitamins that are indispensable to the human body. Vitamin K2 is absorbed and transported through the intestines and transported by the lymphatic system. It requires the participation of bile and pancreatic juice, and is combined with chylomicrons and transported to the liver. In the liver, it is combined with very low-density lipoprotein (VLDL) or low-density lipoprotein (LDL) and transported to various tissues. It is stored in the liver, adrenal glands, lungs, bone marrow, kidneys and other organs, and is finally excreted from the urine in the form of glucuronic acid derivatives.

[0003] Studies have shown that vitamin K2 is beneficial for the treatment of osteoporosis, cardiovascular calcification, cognitive diseases, inflammation and diabetes. Vitamin K2 can activate coagulation factors through post-translational modification of protein molecules, play a role in bone metabolism, and effectively prevent fractures. MK-7 has a longer side chain and higher activity, can exist in the human body for a longer time, and has a higher bioavailability in extrahepatic tissues.

[0004] The basic information of MK-7 is as follows:

[0005] Molecular weight: 649.00; Molecular formula: C 46 H 64 O2; melting point: 54°C; solubility: slightly soluble in chloroform, ethyl acetate, methanol; the structural formula is shown below:

[0006]

[0007] Osteoporosis (OP) is one of the common metabolic bone diseases, characterized by decreased bone density, deterioration of bone tissue microstructure, increased bone fragility and susceptibility to fractures. Osteoporosis is mainly divided into primary osteoporosis and secondary osteoporosis. Primary osteoporosis mainly includes postmenopausal osteoporosis and senile osteoporosis, while secondary osteoporosis is mainly caused by certain diseases or drugs.

[0008] At present, the main therapeutic drugs for osteoporosis include bisphosphonates, estrogen and selective estrogen receptor activators, calcitonin, RANKL monoclonal antibodies, recombinant human parathyroid hormone, etc. However, the above drugs have problems such as poor compliance, obvious side effects, short duration of action, and high price.

[0009] Chinese patent CN110584121A discloses a health product for preventing and treating osteoporosis and muscle cramps and its preparation method. It is a product with vitamin B3 (niacinamide), vitamin D3, vitamin K2 (MK-7), and calcium (calcium carbonate) as core raw materials, with other auxiliary materials, and prepared into tablets after mixing. The invention uses MK-7 type vitamin K2 obtained by fermentation, which has stronger biological activity, higher bioavailability, longer-lasting effect, and is safer and more effective. Vitamin K2 and vitamin D3 work synergistically to participate in bone metabolism, promote the body's effective and rapid absorption of calcium, and improve bone health. In addition, compared with commercially available health products, the content of niacinamide is greatly increased within the range specified by the state, which is more suitable for the prevention and treatment of osteoporosis and muscle cramps.

[0010] Chinese patent CN108065405A discloses a health product for improving bone density, and its components include tricalcium phosphate, calcium citrate, vitamin D3 and vitamin K2. The invention provides a health product for improving bone density, which uses a dual calcium source of calcium phosphate and calcium citrate. Calcium phosphate will not cause bloating and hiccups caused by a large amount of carbon dioxide in the stomach like calcium carbonate, and calcium citrate has a high absorption rate and is suitable for consumers to take at any time; MK-7 type vitamin K2 is added, which can activate osteocalcin, effectively allowing osteocalcin to capture calcium ions absorbed by the human body for bone mineralization, and assisting the human body to effectively utilize the ingested calcium; vitamin K2 and D3 produce a synergistic effect, improve bone health, and can prevent osteoporosis caused by inflammation in women during menstruation.

[0011] The preparations containing MK-7 sold on the market are all health foods and the MK-7 health foods prepared by the above patent cannot replace drugs. Health products mainly play a role in regulating body functions and cannot directly prevent or treat osteoporosis, while drugs have therapeutic effects. In addition, the existing technology adds too many active ingredients to health products, resulting in a low content of MK-7, which is not conducive to combined administration. The present invention aims to provide a pharmaceutical composition with better osteoporosis treatment effect, and the prepared pharmaceutical composition has good stability. Summary of the invention

[0012] In order to achieve the above technical objectives, the present invention hereby proposes the following technical solutions:

[0013] In one aspect, the present invention provides a pharmaceutical composition comprising MK-7 and at least one pharmaceutically acceptable excipient;

[0014] The MK-7 is contained in liposome powder;

[0015] The preparation of the liposome powder comprises the following steps:

[0016] (1) mixing MK-7, an emulsifier and water to obtain a premixed solution and an emulsion;

[0017] (2) mixing the emulsion with auxiliary materials to obtain liposome powder;

[0018] The mass ratio of MK-7, emulsifier and water in step (1) is (0.2-10): (0.05-3): (0.1-8).

[0019] In some embodiments, the MK-7 has the following structure:

[0020]

[0021] Further, in step (1), the mass ratio of MK-7, emulsifier and water is 10:(2.5-2.7):(5-8);

[0022] Furthermore, in step (1), the mass ratio of MK-7, emulsifier and water is 10:2.7:5;

[0023] Furthermore, in step (1), the mass ratio of MK-7, emulsifier and water is 10:2.5:8.

[0024] Specifically, the emulsifier in step (1) is selected from one or more of lecithin, sodium carboxymethyl cellulose, tragacanth gum, gum arabic, deacetylated lecithin, polysorbate, soybean lecithin, egg yolk lecithin, poloxamer, gelatin, xanthan gum, apricot gum, and methylcellulose;

[0025] Furthermore, the emulsifier in step (1) is a combination of lecithin and sodium carboxymethyl cellulose or a combination of tragacanth gum and gum arabic.

[0026] Furthermore, the mass ratio of lecithin to sodium carboxymethyl cellulose is 2:0.5.

[0027] Furthermore, the mass ratio of tragacanth gum to gum arabic is 0.2:2.5.

[0028] Specifically, a preservative may be added during the preparation of the emulsion in step (1), and the preservative is selected from one or more of ethylparaben, methylparaben, and sodium benzoate;

[0029] Furthermore, the preservative described in step (1) is ethylparaben.

[0030] The auxiliary material described in step (2) is selected from one or more of maltodextrin, soluble starch, α-cyclodextrin, β-cyclodextrin, hydroxypropyl starch, mannitol, lactose, microcrystalline cellulose, maltitol, erythritol, xylitol, sorbitol, isomalt, sucrose and glucose.

[0031] Furthermore, the auxiliary material in step (2) is a combination of maltodextrin and cyclodextrin.

[0032] Furthermore, in step (2), the mass ratio of maltodextrin to cyclodextrin is (75-93): (7-7.2).

[0033] In some embodiments, the pharmaceutically acceptable excipient is selected from one or more of solvents, diluents, disintegrants, precipitation inhibitors, surfactants, glidants, binders, lubricants, dispersants, suspending agents, isotonic agents, thickeners, plasticizers, emulsifiers, preservatives, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, flavors, sweeteners, ion exchangers, release agents, coating agents, flavoring agents and antioxidants.

[0034] In some embodiments, the dosage form of the pharmaceutical composition includes, but is not limited to: capsules, tablets, powders, granules, lozenges or liquid preparations, etc.

[0035] Those skilled in the art will appreciate that any dosage form that can maintain the biological activity of the active compound in the pharmaceutical composition of the present invention can be used, and can be obtained without inventive effort.

[0036] In some preferred embodiments, the pharmaceutical composition is in the form of capsules, tablets or granules.

[0037] In some more preferred embodiments, the capsule is a soft capsule.

[0038] In some embodiments, the pharmaceutical composition may contain 0.1-99% of the active compound (MK-7) depending on the administration method.

[0039] In another aspect, the present invention provides a pharmaceutical composition comprising MK-7 or a pharmaceutically acceptable excipient thereof;

[0040] The pharmaceutically acceptable excipient is selected from one or more of vegetable oil, cosolvent, diluent, adhesive, and disintegrant;

[0041] The MK-7 is contained in liposome powder.

[0042] In some embodiments, the preparation of the liposome powder comprises the following steps:

[0043] (1) mixing MK-7, an emulsifier and water to obtain a premixed solution and an emulsion;

[0044] (2) Mixing the emulsion with auxiliary materials to obtain liposome powder.

[0045] In some embodiments, the vegetable oil is selected from one or more of corn oil, peanut oil, almond oil, soybean oil, olive oil, edible blended oil, rapeseed oil, walnut oil, sunflower oil, tea oil, sesame oil, cottonseed oil, peach kernel oil, castor oil, hydrogenated oil, hardened oil, and sesame oil.

[0046] In some preferred embodiments, the vegetable oil is corn oil.

[0047] In some embodiments, the co-solvent includes at least one of Tween 80, Tween 60, Span 20, Span 60, Span 80, glyceryl behenate, glyceryl monolinoleate, propylene glycol monocaprylate, propylene glycol monolaurate, glyceryl monooleate, high-purity diethylene glycol monoethyl ether, linoleic acid polyethanol-6 glyceride, lauric acid polyethanol-6 glyceride, medium chain triglycerides, caprylic acid caprylate, caprylic acid caprylate, propylene glycol fatty acid ester, polyglycerol oleate, oleic acid polyethylene glycol glyceride, oleic acid glyceride, medium chain fatty acid triglycerides, glycerol fatty acid esters, glycerol fatty acid esters, soy lecithin, and glyceryl monooleate.

[0048] In some preferred embodiments, the cosolvent is Tween 80.

[0049] In some embodiments, the diluent is selected from one or more of corn starch, sucrose, maltodextrin, lactose, pregelatinized starch, and microcrystalline cellulose;

[0050] In some preferred embodiments, the diluent is microcrystalline cellulose.

[0051] In some embodiments, the binder is selected from one or more of starch slurry, pregelatinized starch, sucrose, methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, povidone, gelatin, polyethylene glycol, and polyvinyl alcohol.

[0052] In some preferred embodiments, the binder is pregelatinized starch.

[0053] In some embodiments, the disintegrant is selected from one or more of silicon dioxide, magnesium stearate, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, and cross-linked sodium carboxymethyl cellulose.

[0054] In some embodiments, the disintegrants are silicon dioxide and magnesium stearate.

[0055] In some embodiments, the solvent includes water.

[0056] In some embodiments, the pharmaceutical composition further comprises at least one of a preservative, a wetting agent, a lubricant, a colorant, a flavoring agent, and a fragrance.

[0057] In some preferred embodiments, the preservative is selected from one or more of methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, butyl hydroxybenzoate, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, calcium sorbate, benzalkonium bromide, chlorhexidine acetate, o-phenylphenol, eucalyptus oil, cinnamon oil, and peppermint oil.

[0058] In some preferred embodiments, the flavoring agent is selected from one or more of sucrose, simple syrup, stevioside, apalastine, glycerol, sorbitol, and mannitol.

[0059] In some preferred embodiments, the colorant is selected from one or more of tartrazine, hematoxylin, lithospermum root, madder root, beetroot red, cochineal red, turmeric, gardenia, carotene, amaranth, and carmine.

[0060] In some preferred embodiments, the fragrance is selected from volatile oils such as lemon, cherry, mint, and synthetic flavors such as apple flavor and banana flavor.

[0061] In some embodiments, the raw materials for preparing the pharmaceutical composition include:

[0062] 1 part MK-7 liposome powder, 10-30 parts vegetable oil and 0.1-0.5 parts cosolvent.

[0063] In some preferred embodiments, the raw materials for preparing the pharmaceutical composition include:

[0064] 1 part of MK-7 liposome powder, 10-30 parts of vegetable oil, 0.1-0.5 parts of co-solvent and 0.001-1 parts of preservative.

[0065] In some preferred embodiments, the raw materials for preparing the pharmaceutical composition include:

[0066] 1 part of MK-7 liposome powder, 10-30 parts of vegetable oil, 0.1-0.5 parts of cosolvent, 1 part of thickener, 0.4-0.6 parts of plasticizer, 1 part of solvent, 0.0005-0.03 parts of sunscreen and 0.00003-0.0075 parts of colorant.

[0067] In some preferred embodiments, the raw materials for preparing the pharmaceutical composition include:

[0068] 1 part of MK-7 liposome powder, 10-30 parts of vegetable oil, 0.1-0.5 parts of cosolvent, 0.001-1 parts of preservative, 1 part of thickener, 0.4-0.6 parts of plasticizer, 1 part of solvent, 0.0005-0.03 parts of sunscreen and 0.00003-0.0075 parts of colorant.

[0069] The pharmaceutical composition is a soft capsule. Those skilled in the art can, with common technical knowledge, arbitrarily add other pharmaceutical excipients or combine with other active pharmaceutical ingredients (in a form in which the active pharmaceutical ingredients are compatible with each other) without affecting the physiological activity of the active pharmaceutical compound.

[0070] In some embodiments, the raw materials for preparing the pharmaceutical composition include:

[0071] 0.1-1 parts of MK-7 liposome powder, 3-5 parts of diluent, 0.5-2 parts of binder and 0.5-2 parts of disintegrant.

[0072] The pharmaceutical composition is a tablet, and those skilled in the art can, with common technical knowledge, arbitrarily add other pharmaceutical excipients or combine with other active pharmaceutical ingredients (in a form in which the active pharmaceutical ingredients are compatible with each other) without affecting the physiological activity of the active pharmaceutical compound.

[0073] In some embodiments, the pharmaceutical composition is prepared from the following raw materials:

[0074] 1-1.4 parts of MK-7 liposome powder, 1.5-4.2 parts of binder, 0.01-1 parts of preservative, 0.01-1 parts of flavoring agent and 0.00003-0.0075 parts of colorant.

[0075] The pharmaceutical composition is a granule, and those skilled in the art can, with common technical knowledge, arbitrarily add other pharmaceutical excipients or combine with other active pharmaceutical ingredients (in a form in which the active pharmaceutical ingredients are compatible with each other) without affecting the physiological activity of the active pharmaceutical compound.

[0076] In yet another aspect, the present invention provides use of any of the aforementioned pharmaceutical compositions in the preparation of a drug for increasing or improving bone density.

[0077] In yet another aspect, the present invention provides use of any of the aforementioned pharmaceutical compositions in the preparation of a medicament for preventing and / or treating osteoporosis, osteomalacia, osteopenia and / or osteoarthritis.

[0078] In another aspect, the present invention provides a method for preparing any of the aforementioned pharmaceutical compositions, when the pharmaceutical composition is a soft capsule, the preparation method comprises the following steps:

[0079] (1) preparing liposome powder;

[0080] (2) Weighing respective weight portions of liposome powder, vegetable oil, cosolvent and preservative to prepare a drug solution;

[0081] (3) weighing gelatin, plasticizer, sunscreen, colorant and water in parts by weight to prepare capsule wall;

[0082] (4) Inject the drug solution into the compressed capsule skin, compress it together with the capsule skin into soft capsules, wash, dry and package.

[0083] When the pharmaceutical composition is a tablet, the preparation method comprises the following steps:

[0084] (1) preparing liposome powder;

[0085] (2) Mix the liposome powder with a diluent, a binder and a disintegrant, and compress the mixture into tablets.

[0086] When the pharmaceutical composition is a granule, the preparation method comprises the following steps:

[0087] (1) preparing liposome powder;

[0088] (2) The liposome powder is mixed with a binder, a preservative, a flavoring agent and a colorant to prepare a soft material, which is sieved to obtain granules and dried to obtain the granules.

[0089] When the pharmaceutical composition is a hard capsule, the preparation method comprises the following steps:

[0090] (1) preparing liposome powder;

[0091] (2) mixing the liposome powder with a binder, a preservative, a flavoring agent and a colorant to prepare a soft material, sieving the granules and drying the granules;

[0092] (3) The granules prepared in step (2) are loaded into capsule shells to obtain the product.

[0093] The preparation of liposome powder in the above step (1) comprises the following steps:

[0094] (1) mixing MK-7, an emulsifier and water to obtain a premixed solution and an emulsion;

[0095] (2) Mixing the emulsion with auxiliary materials to obtain liposome powder.

[0096] The various preparations involved in the present invention can form a composition with vitamin D3 and calcium to exert a synergistic effect. Calcium supplementation can increase bone mass. Vitamin D3 participates in the process of osteoblast synthesis of osteocalcin. Under the dual effects of the promotion of vitamin K2 (MK-7) and the promotion of calcium absorption by the small intestine by vitamin D3, carboxylated osteocalcin is promoted to combine with hydroxyapatite in the bones, promote bone mineralization, increase bone density, and improve bone quality.

[0097] The beneficial effects of the present invention are:

[0098] The present invention creatively prepares MK-7 raw material into liposome powder, which can greatly improve the stability of the active pharmaceutical ingredient MK-7 in the preparation.

[0099] The pharmaceutical composition prepared by the present invention can improve bone density, increase the content of bone formation factor ALP, inhibit the content of bone resorption factor TRACP-5b, and has a good therapeutic effect on postmenopausal osteoporosis and senile osteoporosis. DETAILED DESCRIPTION

[0100] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly used in the field to which the present invention belongs. For the purpose of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.

[0101] Unless the context clearly dictates otherwise, the expressions "a", "an" and "an" as used herein include plural references. For example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art, and so forth.

[0102] As used herein, the term "about" refers to a range of ±20% of the value that follows. In some embodiments, the term "about" refers to a range of ±10% of the value that follows. In some embodiments, the term "about" refers to a range of ±5% of the value that follows.

[0103] Numerical ranges used herein should be understood to include all numbers within the range. For example, a range of 1 to 20 should be understood to include any number, combination of numbers, or subrange from the following group: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0104] As used herein, the term "comprises" or "comprising" means "including but not limited to". The term is intended to be open-ended to specify the presence of any stated features, elements, integers, steps, or components, but does not exclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" encompasses the more restrictive terms "consisting of" and "consisting essentially of". In one embodiment, the term "comprising" as used throughout the application, especially in the claims, may be replaced by the term "consisting of".

[0105] The terms "optional", "either", "any", or "any" as used herein mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. As used herein, "a" and "an" are used herein to refer to one or more than one of the grammatical objects.

[0106] The term "and / or" used herein should be understood to mean any one of the optional items or a combination of any two or more of the optional items.

[0107] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problems or complications, and are commensurate with a reasonable benefit / risk ratio.

[0108] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with embodiment. In the embodiment, if specific conditions are not specified, the conditions are carried out according to normal conditions or manufacturer's recommendations. If all reagents or instruments are not specified by the manufacturer, they are all conventional products that can be purchased commercially. In order to better illustrate the present invention, numerous specific details are provided in the specific embodiments below. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0109] Basic Example 1 Preparation of liposome powder

[0110] S1: Take 10 g of MK-7 raw material, 2 g of lecithin, and 0.5 g of sodium carboxymethyl cellulose, add water to 5 mL and mix and emulsify (the mass ratio of MK-7 raw material, lecithin, sodium carboxymethyl cellulose and water is 10:2:0.5:5) to obtain a premixed solution, and add 0.001 times of ethylparaben to the premixed solution to obtain an emulsion;

[0111] S2: Take 75.3 g of maltodextrin and 7.2 g of cyclodextrin, add the emulsion obtained in S1 to 100 mL, mix and slurry to obtain slurry;

[0112] S3: The slurry obtained in S2 is dried and crushed to obtain liposome powder containing 10% MK-7.

[0113] Basic Example 2 Preparation of liposome powder

[0114] S1: Take 10 g of MK-7 raw material, 0.2 g of lecithin, and 2.5 g of sodium carboxymethyl cellulose, add water to 8 mL and mix and emulsify (the ratio of MK-7 raw material, lecithin, sodium carboxymethyl cellulose and water is 10:0.2:2.5:8), and add 0.01 times of ethyl hydroxybenzoate to obtain an emulsion;

[0115] S2: Take 82.8 g of maltodextrin and 7 g of cyclodextrin, add the emulsion obtained in S1 to 100 mL, mix and slurry to obtain slurry;

[0116] S3: The slurry obtained in S2 is dried and crushed to obtain liposome powder containing 10% MK-7.

[0117] Example 1 Preparation of MK-7 soft capsules

[0118] The MK-7 soft capsules are prepared from the following raw materials, as shown in Table 1:

[0119] Table 1

[0120]

[0121] The soft capsules are prepared by compression method, where the liquid medicine is added into the hopper of the pill making machine, and the soft capsules are pressed together with the pressed capsule skin, washed, dried and packaged. Each soft capsule contains 1-500 mg MK-7, and the soft capsules contain 15 mg MK-7.

[0122] Example 2 Preparation of MK-7-containing tablets

[0123] The preparation of tablets containing MK-7 is shown in Table 2:

[0124] Table 2

[0125] Components weight 10% MK-7 liposome powder (prepared in Basic Example 2) 150g Pregelatinized starch 400g Microcrystalline Cellulose 100g Silicon dioxide 10g Magnesium Stearate 5g

[0126] When preparing the above tablets, the corresponding amount of active ingredients and excipients are uniformly mixed and tableted. Each tablet of the pharmaceutical composition contains 1-500 mg MK-7, and the above tablet contains 15 mg MK-7.

[0127] Example 3 Preparation of MK-7 granules and hard capsules

[0128] The preparation of MK-7 granules is shown in Table 3:

[0129] Table 3

[0130] Components weight 10% MK-7 liposome powder (prepared in Basic Example 2) 150g Sodium Carboxymethyl Cellulose 225g Methyl hydroxybenzoate 7.5g glycerin 7.5g Lemon Yellow 0.0045g

[0131] When preparing the above granules, the corresponding amount of active ingredients and auxiliary materials are uniformly mixed, a soft material is prepared, granulated through a sieve, and dried. Each granule contains 1-500 mg MK-7, for example, the above granules contain 15 mg MK-7.

[0132] Based on the above granules, the granules are filled into suitable empty capsule shells, which are sealed and cleaned to obtain hard capsules, each of which contains 1-500 mg MK-7.

[0133] Comparative Example 1

[0134] The difference between Comparative Example 1 and Example 1 is that the 10% MK-7 liposome powder in the drug solution is replaced with 15 mg MK-7, and the rest is the same.

[0135] Comparative Example 2

[0136] The only difference between Comparative Example 2 and Example 1 is that the preparation method of MK-7 liposome powder is different, specifically, the emulsifier is replaced by sorbitan monolaurate and sucrose stearate, the mass ratio of sorbitan monolaurate and sucrose stearate is 2:0.5, and the rest are the same.

[0137] Comparative Example 3

[0138] The difference between Comparative Example 3 and Example 1 is that the preparation method of MK-7 liposome powder is different, the ratio of MK-7 raw material, emulsifier and water is 12:5:10, and the rest are the same.

[0139] Comparative Example 4

[0140] The difference between Comparative Example 4 and Example 3 is that the 10% MK-7 liposome powder in the drug solution is replaced with 15 mg MK-7, and the rest is the same.

[0141] Example 4: Improvement effect of the MK-7-containing pharmaceutical composition on the experimental model of postmenopausal osteoporosis

[0142] 1 Model construction

[0143] 110 6-month-old Sprague-Dawley (SD) female rats, weighing 280-330g, were divided into 11 groups, with 10 rats in each group. The ovaries on both sides were removed. The rats were fasted but not watered for 12 hours before the operation. The rats were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (0.2ml / 100g). After anesthesia, the rats were fixed on a board in a supine position, and the hair was shaved in the midline surgical incision area of ​​the lower abdomen with a razor. The rats were disinfected with iodine tincture, the abdominal cavity of the rats was opened, the muscle layer and connective tissue were separated layer by layer, the bilateral ovaries were exposed, and the surrounding connected blood vessels and fallopian tube tissues were ligated with silk thread after lifting. The ovaries were removed, the wounds were sutured, and the abdominal wounds were disinfected with alcohol. 1mL of penicillin (80,000 units / mL) was injected intraperitoneally for each rat to prevent infection. In the sham operation group, fat was removed after laparotomy and the ovaries were retained. The rest of the operations were the same.

[0144] 2 Experimental groups and drug administration

[0145] Rats were randomly divided into sham operation group 1, sham operation group 2, model group 1, model group 2, Example 1-3 groups, and Comparative Example 1-4 groups, with 10 rats in each group. The modeling lasted for 12 weeks. From the 13th week, sham operation group 1 and model group 1 were gavaged with soybean oil (the gavage volume was the same as in Example 1), the soft capsules prepared in Example 1 were broken open, the contents were taken out, and dissolved in 2 ml of soybean oil. Comparative Example 1-3 groups were respectively administered with the pharmaceutical compositions prepared in Comparative Example 1-3 groups, the gavage method was the same as in Example 1, once a day, and the gavage amount was 6 mg / kg / d, for a total of 12 weeks.

[0146] Sham operation group 2 and model group 2 were gavaged with normal saline, and Example 2-3 groups were respectively administered with tablets prepared in Example 2 and granules prepared in Example 3. The drugs prepared in Examples 2 and 3 were crushed and dissolved in normal saline, and the comparative example 4 group was administered with the pharmaceutical composition prepared in comparative example 4, the drugs were crushed and dissolved in normal saline, once a day, and the gavage amount was 6 mg / kg / d, and the administration was given for a total of 12 weeks.

[0147] 3 indicators detection

[0148] The bone density of rats in each group was detected at 12 weeks and 24 weeks of intervention. Dual-energy X-ray bone density analyzer was used to obtain bone density scanning images and measure the femur bone density (BMD) of rats in each group.

[0149] After the rats were killed, blood was collected from the orbits and centrifuged at 3000 rpm for 15 min to separate the serum. The bone formation factor ALP and the bone resorption factor TRACP-5b in the rat serum were detected using an enzyme-linked immunosorbent assay kit according to the instructions of the kit. The detection kit was purchased from Beijing Solebow Co., Ltd.

[0150] 4 Experimental results

[0151] The bone density results of rats in each group are shown in Tables 4-5 below. It can be seen from the table that compared with the sham operation group, the bone density of the model group was significantly reduced. Compared with the model group, the Example 1-3 groups were able to significantly increase the bone density of rats, with statistically significant differences (p < 0.05), indicating that the pharmaceutical composition of the present invention can effectively improve osteoporosis, but there was no significant difference between the comparative example 1-4 groups and the model group (p > 0.05).

[0152] Table 4 Bone density of rats in each group

[0153] Group <![CDATA[Bone density (g / cm 2 )]]> Sham operation group 1 0.1813±0.0035 Model 1 set 0.1421±0.0029** Example 1 Group 0.1745±0.0047## Comparative Example 1 0.1518±0.0052 Comparative Example 2 0.1487±0.0043 Comparative Example 3 0.1495±0.0038

[0154] Note: Compared with the sham operation group, **p<0.01; compared with the model group, ##p<0.01.

[0155] Table 5 Bone density of rats in each group

[0156]

[0157]

[0158] Note: Compared with the sham operation group, **p<0.01; compared with the model group, #p<0.05.

[0159] The detection results of bone formation factor ALP and bone resorption factor TRACP-5b in rat serum are shown in Tables 6 and 7 below. It can be seen that compared with the sham operation group, the ALP content in the model group was significantly reduced, and the TRACP-5b content was significantly increased. The pharmaceutical composition of the administration examples 1-3 groups can increase the content of bone formation factor ALP and significantly reduce the content of bone resorption factor TRACP-5b, indicating that the pharmaceutical composition of the present invention can promote bone formation, inhibit bone resorption, and has a good improvement effect on osteoporosis.

[0160] Table 6

[0161] Group ALP (ng / mL) TRACP-5b (ng / mL) Sham operation group 1 14.76±0.47 5.44±0.25 Model 1 set 12.35±0.61** 6.37±0.58* Example 1 Group 14.53±0.50## 5.61±0.37# Comparative Example 1 12.86±0.70 6.24±0.39 Comparative Example 2 12.53±0.81 6.12±0.70 Comparative Example 3 12.75±1.01 6.18±0.61

[0162] Note: Compared with the sham operation group, *p<0.05, **p<0.01; compared with the model group, #p<0.05, ##p<0.01.

[0163] Table 7

[0164] Group ALP (ng / mL) TRACP-5b (ng / mL) Sham operation group 2 14.28±0.62 5.28±0.35 Model 2 Group 12.17±0.55** 6.88±0.71* Example 2 Group 14.35±0.57## 4.86±0.58## Example 3 Group 14.02±0.73## 5.01±0.64# Comparative Example 4 12.74±0.92 6.38±0.57

[0165] Note: Compared with the sham operation group, *p<0.05, **p<0.01; compared with the model group, #p<0.05, ##p<0.01.

[0166] Example 5 Effect of MK-7-containing pharmaceutical composition on osteoporosis in the elderly

[0167] 1 Model construction

[0168] 110 18-month-old SD male rats, weighing 280-300 g, were randomly divided into normal group 1, normal group 2, model group 1, model group 2, Example 1-3 groups, and comparative example 1-4 groups, with 10 rats in each group. Except for normal groups 1-2, rats in other groups were given dexamethasone sodium phosphate injection by intramuscular injection of the hind limb buttocks at a dose of 2.5 mg / kg, twice a week for 9 consecutive weeks.

[0169] 2 Experimental animal grouping and drug administration

[0170] Rats were randomly divided into normal group 1, normal group 2, model group 1, model group 2, Example 1-3 groups, and comparative example 1-4 groups, with 10 rats in each group. The modeling lasted for 9 weeks. From the 10th week, normal group 1 and model group 1 were gavaged with soybean oil (the gavage volume was the same as in Example 1), and Example 1 group was administered with the soft capsule prepared in Example 1, the soft capsule prepared in Example 1 was broken open, the contents were taken out, and dissolved in soybean oil, and comparative example 1-3 groups were respectively administered with the pharmaceutical composition prepared in comparative example 1-3 groups, the gavage method was the same as in Example 1, once a day, the gavage amount was 6 mg / kg / d, and the administration lasted for 9 weeks.

[0171] Normal group 2 and model group 2 were gavaged with normal saline, Example 2-3 groups were administered with tablets prepared in Example 2 and granules prepared in Example 3, respectively, the drugs prepared in Examples 2 and 3 were crushed and dissolved in normal saline, and the comparative example 4 group was administered with the pharmaceutical composition prepared in comparative example 4, the drugs were crushed and dissolved in normal saline, once a day, the gavage amount was 6 mg / kg / d, and the drug was administered for a total of 9 weeks.

[0172] 3. Detection indicators

[0173] The bone density of rats was tested in the same manner as in Example 3.

[0174] 4 Experimental results

[0175] The results of the bone density test are shown in Tables 8 and 9 below.

[0176] Table 8 Bone density of rats in each group

[0177] Group <![CDATA[Bone density (g / cm 2 )]]> Normal 1 group 0.274±0.019 Model 1 set 0.248±0.025* Example 1 Group 0.281±0.031## Comparative Example 1 0.251±0.022 Comparative Example 2 0.258±0.015 Comparative Example 3 0.253±0.018

[0178] Note: Compared with the normal group, *p<0.05; compared with the model group, ##p<0.01.

[0179] Table 9 Bone density of rats in each group

[0180] Group <![CDATA[Bone density (g / cm 2 )]]> Normal 2 groups 0.281±0.022 Model 2 Group 0.252±0.017* Example 2 Group 0.277±0.025# Example 3 Group 0.283±0.013# Comparative Example 4 0.255±0.025

[0181] Note: Compared with the normal group, *p<0.05; compared with the model group, #p<0.05.

[0182] As can be seen from the above table, compared with the normal group, the bone density of the model group was significantly reduced. Compared with the model group, Example 1-3 groups were able to significantly increase the bone density of rats and effectively improve osteoporosis, but the comparative example 1-4 groups had poor therapeutic effects on osteoporosis.

[0183] Example 6 Preparation stability test

[0184] Experimental methods:

[0185] (1) Influencing factor test

[0186] The preparations prepared by the methods of Examples 1-3 and Comparative Examples 1-4 were placed in high temperature (60°C), high humidity (92.5% RH), and strong light (5500 Lx) environments, respectively, and the effects of the three conditions of high temperature (60°C), high humidity (92.5% RH), and strong light (5500 Lx) on the preparations were tested at different times (0, 5, and 10 days).

[0187] Detection method:

[0188] 1. Determination of MK-7 and MK-6:

[0189] 1. Chromatographic conditions:

[0190] Chromatographic column: C18 column (4.6 mm × 100 mm, 2.6 μm); model: Welch ultimate XB_C18;

[0191] Column temperature: 25°C;

[0192] Detection wavelength: 268nm;

[0193] Injection volume: 10 μl;

[0194] Flow rate: 0.7ml / min;

[0195] Mobile phase: anhydrous ethanol-water = 97:3.

[0196] 2. Solution preparation:

[0197] Preparation of standard solution: Accurately weigh 20 mg of MK-7 standard into a 50 ml volumetric flask, add 1 ml of tetrahydrofuran, and dilute to the mark with anhydrous ethanol; then transfer 1.0 ml to a 10 ml volumetric flask, dilute to the mark with anhydrous ethanol, and mix well.

[0198] Sample solution preparation: transfer an appropriate amount of sample (equivalent to 1 mg of MK-7) to a 25 ml volumetric flask, add 2 ml of tetrahydrofuran, dilute to volume with anhydrous ethanol, shake for 2 minutes, and filter with a membrane with a pore size of 0.45 μm.

[0199]

[0200] Where:

[0201] r U7 ——Peak area of ​​MK-7 peak of sample solution;

[0202] r s ——Average peak area of ​​standard solution 1MK-7 peak;

[0203] C s ——Concentration of standard solution 1MK-7, μg / mL;

[0204] W——sample weight, g;

[0205] V——volume of sample solution, mL.

[0206] The relative retention times of MK-6 and MK-7 were 0.81 and 1.00, respectively.

[0207]

[0208] Where: r U6 ——Peak area of ​​MK-6 peak of test solution;

[0209] r S ——The average value of the main peak area in 6 injections of VK control solution 1;

[0210] Ws——VK reference sample weight, mg;

[0211] W——test sample weight, mg;

[0212] P——labeled content of VK reference substance, %.

[0213] The impurities except MK-6 (relative retention time is about 0.9, calculated in the assay procedure) were calculated using the peak area normalization method.

[0214] Total impurities are the sum of all impurities, including MK-6.

[0215] Table 10

[0216]

[0217]

[0218] (2) Accelerated test

[0219] The preparations prepared by the methods of Examples 1-3 and Comparative Examples 1-4 were placed in an environment of 40°C ± 2°C, 75% RH ± 5% RH (relative humidity) for 0, 1, 2, 3, and 6 months, respectively, and the stability was observed.

[0220] Table 11

[0221]

[0222] (3) Long-term stability test

[0223] The preparations prepared by the methods of Examples 1-3 and Comparative Examples 1-4 were placed in an environment of 25°C ± 2°C, 60% RH ± 5% RH (relative humidity) for 0, 3, 6, 9, 12, 18, and 24 months, respectively, and the stability was observed.

[0224] Experimental results:

[0225] Table 12

[0226]

[0227]

[0228] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises MK-7 and at least one pharmaceutically acceptable excipient; The MK-7 is contained in liposome powder; The preparation of the liposome powder comprises the following steps: (1) Mixing MK-7, an emulsifier and water to obtain a premixed solution and an emulsion; (2) mixing the emulsion with the excipients to obtain liposome powder; In step (1), the mass ratio of MK-7, emulsifier and water is 10:(2.5-2.7):(5-8); The emulsifier in step (1) is selected from a combination of lecithin and sodium carboxymethyl cellulose; The mass ratio of lecithin to sodium carboxymethyl cellulose is 2:0.5; The auxiliary material in step (2) is a combination of maltodextrin and cyclodextrin; The mass ratio of maltodextrin to cyclodextrin in step (2) is (75-93): (7-7.2).

2. The pharmaceutical composition according to claim 1, characterized in that The dosage form of the pharmaceutical composition is capsule, tablet, powder, granule, lozenge or liquid preparation.

3. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises MK-7 and pharmaceutically acceptable excipients; The pharmaceutically acceptable excipient is selected from one or more of vegetable oil, cosolvent, diluent, adhesive, and disintegrant; The MK-7 is contained in liposome powder; The preparation of the liposome powder comprises the following steps: (1) Mixing MK-7, an emulsifier and water to obtain a premixed solution and an emulsion; (2) mixing the emulsion with the excipients to obtain liposome powder; In step (1), the mass ratio of MK-7, emulsifier and water is 10:(2.5-2.7):(5-8); The emulsifier in step (1) is selected from a combination of lecithin and sodium carboxymethyl cellulose; The mass ratio of lecithin to sodium carboxymethyl cellulose is 2:0.5; The auxiliary material in step (2) is a combination of maltodextrin and cyclodextrin; The mass ratio of maltodextrin to cyclodextrin in step (2) is (75-93): (7-7.2).

4. The pharmaceutical composition according to claim 3, characterized in that The vegetable oil is selected from one or more of corn oil, peanut oil, almond oil, soybean oil, olive oil, edible blended oil, rapeseed oil, walnut oil, sunflower oil, tea oil, sesame oil, cottonseed oil, peach kernel oil, hydrogenated oil, hardened oil, and sesame oil; And / or, the cosolvent comprises at least one of Tween 80, Tween 60, Span 20, Span 60, Span 80, glyceryl behenate, glyceryl monolinoleate, propylene glycol monocaprylate, propylene glycol monolaurate, glyceryl monooleate, high-purity diethylene glycol monoethyl ether, linoleic acid polyethanol-6 glyceride, lauric acid polyethanol-6 glyceride, medium chain triglycerides, caprylic acid caprylate, caprylic acid caprylate propylene glycol, propylene glycol fatty acid ester, polyglycerol oleate, oleic acid polyethylene glycol glyceride, oleic acid glyceride, medium chain fatty acid triglycerides, glycerol fatty acid esters, glycerol fatty acid esters, soybean lecithin, and glyceryl monooleate; And / or, the diluent is selected from one or more of corn starch, sucrose, maltodextrin, lactose, pregelatinized starch, and microcrystalline cellulose; And / or, the binder is selected from one or more of starch slurry, pregelatinized starch, sucrose, methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, povidone, gelatin, polyethylene glycol, and polyvinyl alcohol; And / or, the disintegrant is selected from one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, and cross-linked sodium carboxymethyl cellulose.

5. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition further comprises at least one of a preservative, a wetting agent, a lubricant, a colorant, a flavoring agent, and a fragrance.

6. The pharmaceutical composition according to claim 3, characterized in that The raw materials for preparing the pharmaceutical composition include: 1 part MK-7 liposome powder, 10-30 parts vegetable oil and 0.1-0.5 parts cosolvent; or; 1 part MK-7 liposome powder, 10-30 parts vegetable oil, 0.1-0.5 parts cosolvent and 0.001-1 parts preservative; or; 1 part of MK-7 liposome powder, 10-30 parts of vegetable oil, 0.1-0.5 parts of cosolvent, 0.001-1 parts of preservative, 1 part of thickener, 0.4-0.6 parts of plasticizer, 1 part of solvent, 0.0005-0.03 parts of sunscreen and 0.00003-0.0075 parts of colorant; or; 0.1-1 parts of MK-7 liposome powder, 3-5 parts of diluent, 0.5-2 parts of binder and 0.5-2 parts of disintegrant; or; 1-1.4 parts of MK-7 liposome powder, 1.5-4.2 parts of binder, 0.01-1 parts of preservative, 0.01-1 parts of flavoring agent and 0.00003-0.0075 parts of colorant.

7. Use of the pharmaceutical composition according to any one of claims 1 to 6 in the preparation of a drug for increasing or improving bone density.

8. Use of the pharmaceutical composition according to any one of claims 1 to 6 in the preparation of a medicament for preventing and / or treating osteoporosis, osteomalacia and / or osteopenia.

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