CN102670501B - Orally taken vitamin K1 lipid emulsion - Google Patents

Abstract

The invention relates to an orally taken vitamin K1 lipid emulsion, in particular to an emulsion compound. The orally taken vitamin K1 lipid emulsion comprises 0.5-2 percent of vitamin K1, 5-15 percent of soybean oil, 0.5-2 percent of phospholipid, 1-5 percent of low alcohol, 1-5 percent of saccharides and water. The invention also relates to a method for preparing the emulsion compound. The vitamin K1 emulsion compound provided by the invention meets the properties of common pharmaceutics, can be orally administered, is enough in bioavailability and can be suitable for a wider range of people including infants.

Description

The vitamin K1 lipomul that can orally use

Technical field

The invention belongs to medical technical field, relate to a kind of lipomul, particularly relate to a kind of vitamin K1 lipomul that orally uses, this Emulsion can orally use.The invention further relates to the preparation method of preparation lipomul of the present invention.

Background technology

Vitamin K1 is the thick liquid of a kind of Yellow-to-orange clarification, and odorless or odorless are almost met light and easily decomposed.Vitamin K1 is easily molten in chloroform, ether or vegetable oil, and is slightly molten in ethanol, insoluble in water.The chemical name of vitamin K1 is: 2-methyl-3-(3,7,11,15-tetramethyl-2-hexadecene base)-1, and the mixture of the trans and cis-isomer of 4-naphthalenedione, its molecular formula: C31H46O2, molecular weight: 450.71, structural formula is:

Vitamin K1 is a kind of fatsoluble vitamin, extensively is present in the green plants, exist with the form of phylloquinone (phylloquinone), but also synthetic.Vitamin K1 is the yellow oily liquid of coming out of purifying from alfalfa the earliest, and the called after vitamin K1, and people just utilize vitamin K1 to do hemorrhage very early.The medical value of its hemostasis was found in the thirties in 20th century.The conspicuous Garrick of the biochemist of Denmark reaches the disease of nurse research chicken internal hemorrhage, he finds chicken haemophilia when injured, can't condense, and adds certain feedstuff in food, found afterwards to be rich in a kind of material that can stop blooding immediately in this feedstuff, the material of this hemostasis is exactly vitamin K1.After this vitamin K1 is applied to various hemorrhages gradually.Through basic research and clinical use for many years, the pharmacologically active beyond the vitamin K1 hemostasis is found gradually: treat respiratory tract disease, relieving spasm to stop pain, prevent and treat osteoporosis, treat hepatitis, beauty treatment, antitumor etc.Its pharmacologically active is definite, and toxic and side effects is light, so be widely used in clinically, existing vitamin K1 is clinical commonly used drug.

Vitamin K1 is vitamin K class medicine, and vitamin K is liver composition-factor II, VII, IX, the necessary material of X, and vitamin K deficiency can cause these thrombin dyssynthesises or unusual, clinical visible bleeding tendency and prolonged prothrombin.What vitamin K1 can be used for that vitamin K deficiency causes clinically is hemorrhage, as hemorrhage due to obstructive jaundice, leak, the chronic diarrhea etc., vitamin K deficiency in the body due to the Hypoprothrombinemia due to Coumarins, the sodium salicylate etc., prevention hemorrhage of newborn and prolonged application broad ectrum antibiotic.Through basic research and clinical use for many years, find that also vitamin K1 can prevent and treat respiratory tract disease, relieving spasm to stop pain and treatment hepatitis.In addition also with osteoporosis and improve looks relevant.The vitamin K1 pharmacologically active is definite, and poison is secondary light, so be widely used in clinically, now is clinical commonly used drug.

Human body can not synthesize or secrete vitamin K1, and needed vitamin K1 is mainly derived from food, but the escherichia coli in the intestinal also synthetic vitamin K1 be the human body utilization.Vitamin K1 is the main component of forming prothrombing in the blood of human body, can promote liver to make thrombinogen, organize when hemorrhage, platelet is destroyed, discharge thrombokinase and calcium ion, thrombinogen can be combined with thrombokinase and calcium ion, changes thrombin into, change insoluble fibrin into but soluble fibrin is former in the thrombin catalysis blood plasma, reach haemostatic effect.

Vitamin K1 can be removed the smooth muscle spasm contraction, removes oxygen-derived free radicals, and has parahormone sample antiinflammatory action, hinders the release of chemical mediators such as histamine, alleviate the respiratory mucosa inflammation, minimizing is oozed out, blood vessel dilating, improve blood fortune, alleviate heart front and back load, correct heart failure; Excited respiratory center suppresses cerebral cortex, improves brain microcirculation and prevents cerebral edema, is conducive to respiratory failure and corrects, and therefore can alleviate and improve the clinical symptoms of infantile pneumonia.Vitamin K1 can strengthen the effect of some antibiotics in addition, thereby suppresses the pathogen growth, and pertussis is also had curative effect preferably.

Vitamin K1 can directly act on multiple visceral smooth muscle, smooth muscle by lax spasm stops the internal organs pain due to the multiple illness, as biliary colic, renal colic, and ureter and cystolith angor etc., effective percentage can reach 80～90%, and analgesic effect is fast, effect is lasting, safety, curative effect height.

The shortage of vitamin K1 can cause blood not solidify, and mostly is neonate because vitamin K1 lacks hemorrhage patient in clinical.1～2 day diet of neonate birth is limited, the intake of vitamin K is few, vitamin content is low in the breast milk, and is aseptic in the intestinal in addition, influences the synthetic of vitamin K, cause vitamin K deficiency, liver is not mature enough, and liver synthesizes the process of prothrombin, VII, IX, X and is obstructed cruor time extending, cause hemorrhagely, this disease is hemorrhagic disease of the newborn.Generation and the avitaminous relation of hemorrhagic disease of the newborn (HDN) have just been described as far back as nineteen fifty-two Dam etc.Can be divided into three kinds according to disease time: early hair style hemorrhage, typical hemorrhage, delayed.K1 is deficient in vitamin in the body when neonate just has been born, the pure breast feeding time, long neonate can fast-developingly lack to serious vitamin K1: children's can taken place typical or the delayed hemorrhage occurring in 2 months in first week after the birth, and the both of these case morbidity causes that suddenly, easily intracranial hemorrhage damage, disability rate height, mortality rate are also high.The sickness rate of China is up to 0.5～2%, and the sickness rate in area, Yantai is up to 9.03%, and China is the hotspot of vitamin K deficiency hemorrhagic disease.Medical research shows that the neonate birth gives vitamin K1 can prevent vitamin K1 shortage property hemorrhage.The route of administration of vitamin K1 also influences drug effect.Oral vitamin K1 only can prevent typical hemorrhage, and is invalid to the delayed hemorrhage, and Longhan finds that vitamin K1 is sustainable more than 2 months with 2mg intramuscular injection drug effect, oral relatively poor relatively.2000, the intramuscular injection of the interior conventional application vitamin K1 of neonate birth 24h was promoted once in Heng County, China Guangxi, and result of study shows: baby's vitamin K deficiency number of hospitalized obviously reduces, and vitamin K deficiency patient descends.In addition, because the human liver can synthesize thrombinogen, hepatic disfunction person or surgery large operation person all need give a certain amount of vitamin K1 and can prevent hemorrhage.

Infantile respiratory disease is occupied pediatric disease sickness rate 70%, and the winter-spring season sickness rate is higher, mainly shows as pneumonia, asthma, bronchitis, pertussis.

Infantile pneumonia refers to cause the lung microcirculation disturbance by various different pathogens and other caused by factors pneumonias, and reasons such as anoxia, infection, heating make blood present highly enriched, high gathering, high thickness state, can cause and increase the weight of microcirculation disturbance.Vitamin K1 can change (cAMP/cGMP) ratio in the cell, removes the smooth muscle spasm contraction, removes oxygen-derived free radicals, and have parahormone sample antiinflammatory action, and hinder the release of chemical mediators such as histamine, alleviate the respiratory mucosa inflammation, minimizing is oozed out, and reaches the effect that cough-relieving is coughed and relievingd asthma; Vitamin K1 is expansible blood vessel also, improves blood fortune, alleviates heart front and back load, corrects heart failure; Excited respiratory center; suppress cerebral cortex; improve brain microcirculation and prevent cerebral edema; can reduce airway resistance; increase alveolar ventilation; the ventilation ventilation of alleviating crisis life is depleted; be conducive to exhale the correction that declines; therefore the clinical symptoms of infantile pneumonia can be alleviated and improve to vitamin K1, also can reduce the case fatality rate of severe pneumonia simultaneously greatly, shortens the hospital stays; measure nonresponders such as aminophylline, promethazine, dexamethasone, oxygen uptake are used again; still can receive promising result, have the people that severe pneumonia of infants 100 examples are assisted a ruler in governing a country with vitamin K1 and treat 50 examples, obtain effect preferably.

Vitamin K1 treatment bronchiolitis and bronchial asthma report are also many, and domestic have research from 41 routine children asthmatic brouchitis case intravenous applications vitamin K1 treatment asthmatic bronchitiss of institute in October in October, 1998 to calendar year 2001, satisfactory effect.Behind the child vein instillation vitamin K1, infant is quiet rapidly, and very fast sleeping, cough with asthma obviously reduces, and breathes smooth-going.

Vitamin K1 also has curative effect preferably to pertussis.Pertussis is the acute respiratory infectious disease that is caused by bordetella pertussis.Infectiousness is strong, is more common in infant, with liver source property blood coagulation disorders, causes the petechial hemorrhage of respiratory mucosa during the pertussis outbreak, and this produces paroxysmal spasmodic cough the cause of disease, and the respiratory tract petechial hemorrhage is relevant with venous stasis or blood vessel toxic lesion.Though vitamin K1 can not directly act on pathogen, it can strengthen the effect of some antibiotics, thereby suppresses the pathogen growth.

Animal experiment study proved in 1974, the vitamin K1 intestinal that myenteron shrinks automatically and acetylcholine causes to exsomatizing shrinks the obvious suppression effect is arranged, in recent years clinical trial shows: vitamin K1 can directly act on multiple visceral smooth muscle, smooth muscle by lax spasm, and stop internal organs pain due to the multiple illness, as biliary colic, renal colic, ureter and cystolith angor etc., effective percentage can reach 80～90%, and clinical also have good effect with vitamin K1 treatment children's gastrointestinal convulsion stomachache.

Vitamin K1 can be used for treatment or prevention and osteoporosis diseases associated or disease.Nineteen sixty, people such as Bouckaert report that vitamin K can promote the union of fracture of rat and rabbit.Discovery pregnant woman oral anticoagulant such as Pettifor produced baby's nasal bone hypoplasia in 1975, reported first the influence of vitamin K deficiency to the human body bone development, the hypothesis of vitamin K participant bone metabolism; Pettifor thinks that vitimin supplement K can promote bone formation, reduce bone catabolism, osteoporosis there is positive preventive and therapeutic action, its mechanism of action is to be converted into γ-Qiang Jiguansuan residue (Gla) as the glutaminic acid residue in some protein of cofactor catalysis of enzyme, this albuminoid is called vitamin K dependent protein matter again, and the γ-Qiang Jiguansuan residue after its activation has in conjunction with Ca ⁺⁺Characteristic, thereby make vitamin k-dependent protein bring into play its biological activity, work as vitamin K deficiency, therefore this proteinoid hydroxylation obstacle, its biological activity also reduce.More bases and clinical trial have illustrated vitamin K and osteoporotic relation.Other has the recycling clear proof that causes vitamin K deficiency of vitamin K in the anticoagulant body capable of blocking.The clinical report anticoagulant was arranged to the side effect of bone metabolism in 1975, after this crowd and animal experiment constantly are confirmed, behind the long diaphysis bone, and the too early calcification of end growth hone lamella.More than test all points out vitamin K deficiency can disturb the osseous tissue homergy.

Vitamin K1 helps treatment or the prevention of liver inflammatory diseases.Studies confirm that vitamin K1 can strengthen the effect of pathological changes hepatocyte vigor and absorbability.Also have report to adopt megavitamin K " to swash and raise therapy " the treatment hepatopathy, vitamin K can promote the hepatocyte vigor of impending death, strengthens the absorbing power to nutrient substance, accelerates histiocytic recovery.

Version Chinese Pharmacopoeia in 2010 has recorded vitamin K 1 injection (referring to two ones the 910th page of version Chinese Pharmacopoeia in 2010), this is that a kind of principal agent concentration is the aquesterilisa dispersion liquid that is yellow liquid of 10mg/ml, it allows little apparent muddiness, need antifreeze preservation, if oil droplet is arranged separates out or layering, can under the shading condition, be heated to 70-80 ℃, jolting makes its natural cooling, if clarity normally still can continue to use, these information show that the injection of this pharmacopeia record is a kind of injection that uses solubilizing agent that medicine directly is dissolved in the water, rather than the Emulsion of pharmaceutics field common sense because Emulsion generation oil droplet is separated out or layering after can't after being heated to 70-80 ℃ and jolting, can reach clinical acceptable clarity.

WO2011/153513A2 discloses the micro emulsion composition of fatsoluble vitamin K, it is said vitamin K1 injectable emulsion (the Phytonadione Injectable Emulsion of the alternative American Pharmacopeia of this micro emulsion composition, USP, can abbreviate PIE-USP as, its former name is vitamin K 1 injection) for clinical treatment.PIE-USP is a kind of aqueous dispersions, wherein contains polyoxyethylene fatty acid (also being called Cremophor), and this is a kind of potent surfactant.Although nomenclature of drug is recited as the injection breast, the former product that grinds of vitamin K1 injectable emulsion (USP) is Merck ﹠ Co., Inc.

It does not in fact contain any oil, therefore is different from the vitamin K1 lipomul of indication of the present invention.Other manufacturer also uses the surfactant that is similar to Cremophor to come the solubilising vitamin K1, for example uses the HCO-60 of high HLB value.The shortcoming of using this type of surfactant has been described among the WO2011/153513A2, and by not using this type of reagent, but prepare a microemulsion to substitute existing P IE-USP, this microemulsion for example contains 0.2～1% vitamin K1,0.5～2% soybean oil, 0.5～2% MCT, 4～25% phospholipid and 10% sucrose, and phospholipid is 13-25 times of vitamin K1 amount.Yet this microemulsion that constitutes with relatively large phospholipid and more a spot of oil is a kind of metastable state that is in the critical state between solution and the Emulsion, its easily because of outside cause for example pressure sterilizing break away from this metastable state, for example in the preparation embodiment of WO2011/153513A2, need with 0.22 μ m membrane filtration degerming.In addition, contain a certain amount of oil for example soybean oil be favourable, the soybean oil of 10% amount for example, its can for human body provide can't or the energy of inconvenience picked-up.Be what fully to have proved than the safety of oil for injection (for example safflower oil, the Fructus Canarii albi wet goods) soybean oil of other kind and effectiveness, the heavy dose of soybean oil that provides with the fat emulsion injection form has used many decades clinically.

Yet the route of administration of existing vitamin K1 mainly biases toward injection system, and this mode is difficult with respect to oral administration for patient's compliance, but oral administration makes people face the challenge aspect drug bioavailability again.In addition, although existing VK1 tablet applications in clinical, this kind tablet is suitable for for the part adult, for infant, swallow inconvenient patient and inapplicable.

People expect that still new Vitamin K1 preparation is arranged, and for example it meets general pharmaceutics character, can be taken orally and has enough bioavailability again, and going for widely, the crowd for example comprises infant.

Summary of the invention

The objective of the invention is provides a kind of new Vitamin K1 preparation for clinical, and for example it meets general pharmaceutics character, can be taken orally and has enough bioavailability again.The inventor finds, the vitamin K1 Emulsion that is prepared by the special formulation that comprises about 10% soybean oil, it is the fat milk that a kind of performance can satisfy the clinical application demand fully, and it has the bioavailability of desirable when oral administration, for example comprises infant applicable to crowd widely.The present invention is based on this discovery and be accomplished.

For this reason, first aspect present invention provides emulsion composition, and it comprises:

Vitamin K1 0.5-2%,

Soybean oil 5～15%,

Phosphatidase 10 .5～2%,

Lower alcohol 1～5%,

Saccharide 1～5%,

Water is an amount of, adds to 100%.

According to the emulsion composition of first aspect present invention, wherein said lower alcohol is selected from propylene glycol, glycerol or its combination.

According to the emulsion composition of first aspect present invention, wherein said saccharide is selected from sorbitol, mannitol, fructose, glucose or its combination.

According to the emulsion composition of first aspect present invention, wherein also comprise correctives.In one embodiment, described correctives can be selected from aspartame, saccharin sodium, sucralose etc. and their combination.According to the present invention, the purposes of described correctives in the present composition is that those skilled in the art are rule of thumb confirmable, for example 0～1%, for example 0.01～0.5%, for example 0.02～0.2%.For the present composition, whether the existence of correctives is for realizing that the object of the invention does not have substantial influence.

According to the emulsion composition of first aspect present invention, wherein also comprise antiseptic.In one embodiment, described antiseptic can be selected from benzyl alcohol, parabens (for example methyl hydroxybenzoate, ethyl hydroxybenzoate, propylparaben), chlorobutanol etc. and their combination.According to the present invention, the purposes of described antiseptic in the present composition is that those skilled in the art are rule of thumb confirmable, for example 0～1%, for example 0.01～0.5%, for example 0.02～0.2%.For the present composition, whether the existence of antiseptic is for realizing that the object of the invention does not have substantial influence.Certainly, under present composition case of oral administration, Orally taken emulsion can be considered multiple-unit container, and add an amount of antiseptic this moment is useful for the clinical practice of medicine.In addition, under the situation of present composition drug administration by injection, injectable emulsion is advised not adding preservative agent; Because Emulsion of the present invention can be by the mode degerming of pressure sterilizing, therefore adding preservative agent is not fully feasible for the present composition yet.

According to the emulsion composition of first aspect present invention, wherein also comprise the pH regulator agent.In one embodiment, described pH regulator agent can be selected from any acceptable acid of pharmacy or alkali that can be used for regulating pH value, such as but not limited to hydrochloric acid, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, phosphoric acid, sulphuric acid etc.According to the present invention, the purposes of described pH regulator agent in the present composition is that those skilled in the art are rule of thumb confirmable, for example an amount of pH regulator agent of general use makes the pH value of emulsion composition of the present invention be adjusted to the scope of expectation, the for example scope of pH4-9, for example scope of pH5-8.

According to the emulsion composition of first aspect present invention, it comprises:

Vitamin K1 0.75-1.5%,

Soybean oil 8～12%,

Phosphatidase 10 .75～1.5%,

Lower alcohol 2～4%,

Saccharide 3～5%,

Water is an amount of, adds to 100%.

According to the emulsion composition of first aspect present invention, it comprises:

Vitamin K1 0.9-1.1%,

Soybean oil 9～112%,

Phosphatidase 11 .0～1.5%,

Lower alcohol 2～3%,

Saccharide 3～4%,

Water is an amount of, adds to 100%.

According to the emulsion composition of first aspect present invention, it is measured according to the wet method in Pharmacopoeia of People's Republic of China version two appendix IXE granularity in 2010 and the particle size distribution method three therapeutic methods of traditional Chinese medicine (light scattering method), and the particle diameter of the breast 70% or more is below 5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is below 5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is below 5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.01～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.01～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.01～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.05～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.05～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.05～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.1～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.1～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.1～5.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.1～2.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.1～2.0 μ m.In one embodiment, particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter is less than 20% greater than the breast grain of 5 μ m, preferably is less than 15%, is less than 10%, is less than 5%.

Emulsion composition according to first aspect present invention, it is according to 2010 editions two described high performance liquid chromatography of appendix V D of Chinese Pharmacopoeia, measure with following liquid phase chromatogram condition, the amount that contains vitamin K1 among every 1ml be 5～20mg (7.5～15mg for example, for example 9～11mg):

(1) chromatographic condition and system suitability test: be filler with octadecylsilane chemically bonded silica, be mobile phase with 95: 5 alcohol-water mixed liquor of volume ratio, column temperature is 35 ℃, and the detection wavelength is 254nm, and theoretical cam curve is calculated with vitamin K1 should be not less than 1500;

(2) preparation of need testing solution: it is an amount of that precision is measured described compositions, adds the isopropyl alcohol dissolving and quantitatively make to contain the solution that vitamin K1 is about 0.05mg among every 1ml, as need testing solution;

(3) preparation of reference substance solution: it is an amount of to take by weighing the vitamin K1 reference substance, accurately claims surely, and (in one embodiment, this 10% fat milk is according to the present invention hereinafter method of preparation example 1 but 10% fat milk of vitaminize K1 preparation not to add 10% fat milk; In one embodiment, this 10% fat milk is 10% commercially available fat emulsion injection) 1ml, add isopropyl alcohol dissolving and quantitatively make and contain the solution that vitamin K1 is about 0.05mg, product solution in contrast among every 1ml;

(4) algoscopy: get each 20 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, the record chromatogram is by the content of external standard method with vitamin K1 in the described compositions of calculated by peak area.

According to the emulsion composition of first aspect present invention, it is measured with following method, among every 1ml glycerinated amount be 15～30mg (17.5～25mg for example, for example 19.5～24.5mg):

Precision is measured described compositions 2.5ml, puts in the conical flask, adds water 100ml, adds 5 of bromocresol purple indicator solutions, shakes up, if show acid, drips the 0.1mol/L sodium hydroxide solution, makes solution be bluish violet; If show alkalescence, should drip the 0.5mol/L sulfuric acid solution earlier is adjusted to solution and just is yellow, dripping the 0.1mol/L sodium hydroxide solution again makes solution be bluish violet, add the 0.7% Potassium metaperiodate. solution 100ml that faces with newly joining, put in 37～40 ℃ the water-bath insulation 15 minutes, and jolting constantly, add 1,2-propylene glycol 3ml, placed 5 minutes, just be bluish violet with 0.1mol/L sodium hydroxide volumetric solution titration to solution, namely get the amount of used sodium hydroxide volumetric solution, be equivalent to the C3H8O3 calculating glycerol content of 9.210mg with the 0.1mol/L sodium hydroxide volumetric solution of every 1ml.

Emulsion composition according to first aspect present invention, it is according to 2010 editions two described high performance liquid chromatography of appendix V D of Chinese Pharmacopoeia, measure with following liquid phase chromatogram condition, the amount that contains triglyceride among every 1ml be 75～125mg (80～120mg for example, for example 90～110mg):

(1) chromatographic condition and system suitability test: be filler with silica gel, be that normal hexane-isopropyl alcohol-acetic acid mixed liquor of 98.9: 1: 0.1 is mobile phase with volume ratio, evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 ℃) detects, the separating degree of triolein and oleic acid should be greater than 2, and the relative standard deviation of the peak area of triglyceride should be not more than 3.0%;

(2) preparation of system suitability testing liquid: get each 10mg of triglyceride and oleic acid, to the 25ml measuring bottle, with mobile phase dissolving and be diluted to scale, shake up, namely;

(3) preparation of reference substance solution: get the about 0.35g of soybean oil reference substance, the accurate title, decide, put in the 50ml measuring bottle, with appearance mixed solutions such as normal hexane and isopropyl alcohol dissolvings and be diluted to scale, be stock solution, precision is measured stock solution 3ml and 4ml, put respectively in the 25ml measuring bottle, be diluted to scale with mobile phase, shake up, be reference substance solution 1 and reference substance solution 2;

(4) preparation of need testing solution: precision is measured described compositions 4ml, puts in the 50ml measuring bottle, with appearance mixed solutions such as normal hexane and isopropyl alcohol dissolvings and be diluted to scale, shakes up; Precision is measured 3ml, puts in the 25ml measuring bottle, is diluted to scale with mobile phase, shakes up, namely;

(5) algoscopy: precision is measured reference substance solution 1, reference substance solution 2 respectively, and each 10 μ l of need testing solution alternately inject chromatograph of liquid, get calculated by peak area through chromatographic isolation.

According to the emulsion composition of first aspect present invention, it comprises the vitamin K1 of 0.5-2%, preferably comprises the vitamin K1 of 0.75-1.5%, preferably comprises the vitamin K1 of 0.9-1.1%, preferably comprises about 1.0% vitamin K1.

According to the emulsion composition of first aspect present invention, it comprises 7.5～12.5% soybean oil, preferably comprises 8～12% soybean oil, preferably comprises 9～11% soybean oil, preferably comprises about 10% soybean oil.

According to the emulsion composition of first aspect present invention, it comprises 0.8～2% phospholipid, preferably comprises 1.0～1.5% phospholipid, preferably comprises about 1.2% phospholipid.

According to the emulsion composition of first aspect present invention, it comprises 1～5% lower alcohol, preferably comprises 2～4% lower alcohol, preferably comprises 2～3% lower alcohol.In one embodiment, described lower alcohol is glycerol.In one embodiment, described lower alcohol is propylene glycol.In one embodiment, described lower alcohol is the combination of the arbitrary ratio of propylene glycol and glycerol.Owing to use glycerol or propylene glycol in some example of the present invention separately, and be used in combination propylene glycol and glycerol with certain ratio in some example, so those skilled in the art can expect fully propylene glycol and glycerol the two all can be used for the present invention with the combination of any ratio.

Emulsion composition according to first aspect present invention, it comprises 1～5% saccharide, for example comprise 2～5% saccharide, for example comprise 3～5% saccharide, for example comprise 4～5% saccharide, for example comprise 1～4% saccharide, for example comprise 1～3% saccharide, for example comprise 1～2% saccharide, for example comprise 2～4% saccharide, for example comprise 2～3% saccharide, for example comprise 3～4% saccharide.In one embodiment, described saccharide is sorbitol.In one embodiment, described saccharide is monosaccharide.In one embodiment, described saccharide is mannitol.In one embodiment, described saccharide is fructose.In one embodiment, described saccharide is glucose.In one embodiment, described saccharide is any two or more the combination in sorbitol, mannitol, fructose, the glucose.Owing to use sorbitol, mannitol, fructose or glucose in some example of the present invention separately, and in some example with certain ratio be used in combination in sorbitol, mannitol, fructose, the glucose any two or more, so those skilled in the art can expect fully in sorbitol, mannitol, fructose, the glucose any two or more all can be used for the present invention with any ratio combination.

According to the emulsion composition of first aspect present invention, it is oil-in-water emulsion.The emulsion composition according to the present invention, it is Emulsion for oral use.The emulsion composition according to the present invention, it is injectable emulsion.

Second aspect present invention provides the method for preparing the described emulsion composition of first aspect present invention, and it may further comprise the steps:

(1) take by weighing vitamin K1, soybean oil, phospholipid, lower alcohol, saccharide by recipe quantity, and other optional material, standby;

(2) water (for example purified water, water for injection) that will prepare the about 60-90% of total amount (for example about 80%) adds in the Agitation Tank, adds the lower alcohol of recipe quantity, stirs, and makes water;

(3) soybean oil with recipe quantity adds in another Agitation Tank, feeds nitrogen current, adds vitamin K1 and the phospholipid of recipe quantity, is stirred to each material dissolution, makes oil phase;

(4) feed nitrogen current, oil phase is slowly added the aqueous phase of high-speed stirred, it is in good time to continue high-speed stirred, regulates its pH value to prescribed limit with sodium hydroxide solution, gets colostrum;

(5) under nitrogen current, make colostrum high pressure homogenize under 40-70MPa (for example 55-60MPa) pressure, low pressure homogenize under 5-25MPa (for example 12-18MPa) pressure is cooled off again;

(6) add saccharide with an amount of water-soluble recipe quantity, add water again and be settled to theoretical recipe quantity, stir, in case of necessity with sodium hydroxide or hydrochloric acid solution regulator solution pH value to prescribed limit, use the membrane filtration of 10 μ m then;

(7) the filtrate fill is in drug packaging container (vial (for example cillin bottle, ampoule bottle, phial for example, and the injecting drug use packaging vial of other type or the oral medication packaging vial of other type etc.), plastic bottle (for example plastic ampoule, plastics oral liquid bottle, and the injecting drug use packaging plastic bottle of other type or the oral medication packaging plastic bottle of other type etc.) in, sealing, randomly carry out pressure sterilizing, namely.

According to the method for second aspect present invention, it is included in the suitable processing step, adds other optional adjuvants.In one embodiment, in step (3), add stabilizing agent with vitamin K1 and phospholipid.In one embodiment, in step (4), must add isotonic agent behind the colostrum.

According to the method for second aspect present invention, if also comprise other adjuvant in the emulsion composition, for example coemulsifier, stabilizing agent etc., these adjuvants can add at the preparation water or at the phase time that makes up oil according to its hydrophilic or lipophile.

According to the method for second aspect present invention, before compositions encapsulation and sterilization, measure and or the pH value of regulating said composition be to be easy to very much; And (namely becoming whole breast) should not be regulated the pH value of said composition usually again after compositions encapsulation and sterilization.Those skilled in the art understand, the pH value of compositions may change to some extent before and after sterilization, perhaps increase a little or reduce a little, therefore when in the dosing process, pH value being regulated, the pH value that can suitably depart from objectives makes pH value after the compositions sterilization fall into as far as possible or near purpose pH value or scope.In this sense, in the present invention, when mentioning the pH value of emulsion composition, if not explanation in addition refers to sterilization treatment pH value afterwards; For example hereinafter in the preparation example 1, when describing composite formula, mention " the pH regulator agent is an amount of; adjust pH to 8.0 " refer in prescription or the dosing process in to add an amount of pH regulator agent an amount of, make medicinal liquid through corresponding preparation method processing and after pressure sterilizing obtains final emulsion composition, the pH value of this emulsion composition is about 8.0, and for example pH value is in 8.0 ± 0.2 scope, and for example pH value is in 8.0 ± 0.1 scope.

The documents and materials of the present invention's citation, its full content is incorporated this paper by reference into.In either side of the present invention, any one of its arbitrary embodiment or a plurality of technical characterictic can be combined in arbitrary other embodiment of this aspect, also can be combined in arbitrary embodiment on the other hand, as long as this combination contradiction can not occur.

The various terms that the present invention uses have the conventional general sense of understanding of those skilled in the art, when inconsistent with this general sense, are as the criterion with the present invention.

In the present invention, for writing conveniently, vitamin K1 can simply be written as VK1.

The present invention provides a kind of fluid composition generally, and main carrier wherein is water, and solute comprises liquid substance (for example lower alcohol) and/or solid matter (Ovum Gallus domesticus Flavus lecithin and the saccharide that for example are semi-solid).Therefore, for simplicity, in the present invention, when under the situation of indicated concentration percent, using " % ", as not explanation in addition, refer to weight/volume percent; For example.For the prescription that contains following ratio: vitamin K1 is 1%, soybean oil 10%, Ovum Gallus domesticus Flavus lecithin 1.2%, glycerol 2.2%, mannitol 3%, water add to 100% in right amount, and it refers to that containing vitamin K1 in every 100ml prescription is 1g, soybean oil 10g, Ovum Gallus domesticus Flavus lecithin 1.2g, glycerol 2.2g, mannitol 3g and the water that adds to the 100ml volume.

It will be appreciated by those skilled in the art that breast grain particle diameter normally statistical way calculate or provide.For example, phrase " particle diameter of the breast grain more than 70% is below 5.0 μ m " can refer to measure according to (for example microscopic method, light scattering method etc.) someway the particle diameter of the breast grain particle of some, for example measure about 1000 breast grain particles, then particle diameter is dropped on population in the following scope of 5.0 μ m divided by these about 1000 populations of statistical computation, the percent that obtains is greater than 70%.Other similar terms also has similar meaning.

In the present invention, phrase " breast grain particle diameter is about 1 μ m " as not explanation in addition, refers to that the particle diameter of breast grain is between 0.5 μ m～2.0 μ m.

In the present invention, term " 10% fat emulsion injection " as not explanation in addition, but refers to according to method of preparing emulsion, basically by comprising the intravenous injection that following formulation obtains or the Emulsion that orally uses:

Soybean oil 10%,

Ovum Gallus domesticus Flavus lecithin 1.2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%.

Above-mentioned " 10% fat emulsion injection " can be used as the contrast reagent of measuring emulsion composition of the present invention.

Emulsion composition of the present invention be can be taken orally or inject intravenous administration particularly; Be used under the case of oral administration in expection, wherein each kind of material is preferably preparing burden by rank material for oral use; Be used under the situation of drug administration by injection in expection, wherein each kind of material preferably prepared burden with injection rank material.In emulsion composition of the present invention, vitamin K1 exists as principal agent or active component.In emulsion composition of the present invention, soybean oil uses as the oil phase solvent usually, and it also has the effect that energy is provided certainly.Usually bring into play the effect of emulsifying agent at emulsion composition mesolecithal lecithin of the present invention, and lower alcohol can be brought into play the effect of osmotic pressure regulator usually; Further, water for injection uses as the water solvent.Yet the inventor finds, is used under the situation of Emulsion of the present invention at lower alcohol and saccharide combination, and this emulsion composition has good galenic pharmacy characteristic.

Undoubtedly, the adjuvant that the present invention is used comprises soybean oil, is to meet necessarily requiredly, and for example, during as fat emulsion for injection, these adjuvants should use for injection in preparation; And when preparing as lipomul for oral use, the prescription of these adjuvants is lower.But the difference of these injections of adjuvant itself and prescription for oral use does not have substantial harmful effect for the galenic pharmacy performance of Emulsion of the present invention, and this is well known to a person skilled in the art.

Phospholipid of the present invention is to be selected from following refining phospholipid: Ovum Gallus domesticus Flavus lecithin, soybean lecithin, cephalin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline etc.They can be bought from the market, and also available usual method is prepared as organic solvent segregation.Just raw phospholipid is dissolved in cold normal hexane-acetone, stirs down and slowly adds acetone, the filtered and recycled insoluble matter, repeat this operation after, steam solvent and just can obtain refined phosphatide.Its Main Ingredients and Appearance comprises phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE etc.As other phospholipid also can give an example out phosphatidylinositols, Phosphatidylserine, sphingomyelins etc.The phospholipid of indication of the present invention is Ovum Gallus domesticus Flavus lecithin preferably.

The present invention contains in the lipomul of vitamin K1, can also further comprise pharmaceutically acceptable adjuvant as required, as coemulsifier, stabilizing agent, osmotic pressure regulator, pH value regulator etc.

Can be that the carbon number that contains 50g or following amount in every 1000ml lipomul is 6～22 as coemulsifier, preferred carbon number is 12～20 fatty acid or its physiologically acceptable salt, this fatty acid can be straight chain, it also can be side chain, the preferably stearic acid of straight chain, oleic acid, linoleic acid, Palmic acid, linolein acid, tetradecanoic acid etc., can enumerate physiologically acceptable salt as these sour salt, as sodium salt, potassium salt, calcium salt etc.

Can also be the medium chain fatty glyceride that contains 50g or following amount in every 1000ml lipomul as coemulsifier, said medium chain fatty glyceride be that C is arranged ₈～C ₁₂An ester, diester or three esters that the fatty acid of chain length and glycerol form, or the mixture of these esters, the preferably triglyceride of the sad and capric acid of satisfied fatty acid, or its commercial product is as Miglyol 812 etc.

In every 1000ml lipomul, contain 10g or following amount as stabilizing agent, the cholesterol of preferred 5g or following amount or phospholipid acid etc.

Osmotic pressure regulator is optional from albumin, dextrin, ethene polymers, non-ionic surface active agent, gelatin, hetastarch, propylene glycol, glycerol etc., making the osmotic pressure ratio (assay method carries out according to " Chinese Pharmacopoeia version in 2010 two appendix IX G " osmotic pressure molar density algoscopy " ") of the lipomul of making is 0.5～3, and preferred osmotic pressure ratio is 0.8～1.5.

In the present invention, mention " Chinese Pharmacopoeia ", refer to Pharmacopoeia of People's Republic of China, as not dated especially, refer to the version of version in 2010; As not dated especially, refer to the related content of two ones of versions in 2010.

The pH regulator agent that the present invention uses includes but not limited to sodium hydroxide solution, hydrochloric acid solution, for example 1M sodium hydroxide solution, 1M hydrochloric acid solution.In the present invention, when mentioning emulsion composition of the present invention or being used for the pH value of control composition, this pH value is the final pH value of compositions, rather than the pH value of its middle material.For example for Emulsion of the present invention, the pH value of measuring before branch installs in the packing container might have slight variations with the pH value that sterilization is measured afterwards, the pH value of Emulsion after the pH value of mentioning among the present invention refers to sterilize.So in production practices, when before sterilization, preparing burden, the possible variation of pH value before and after this sterilization with due regard to; If for example pH value can reduce by 0.2 unit after sterilization, and the target pH value is 8.0, the pH value of the emulsion composition of preparing before then sterilizing is adjusted to 8.2 and is advisable.

Lipomul provided by the invention can prepare by the following method:

Take by weighing vitamin K1, soybean oil, phospholipid, lower alcohol, saccharide by recipe quantity, standby; The water of preparation total amount about 80% is added stainless cylinder of steel, add the lower alcohol of recipe quantity while stirring; The soybean oil of recipe quantity is added in another stainless cylinder of steel, feed nitrogen current, add vitamin K1 and the phospholipid of recipe quantity, be stirred to the phospholipid dissolving; Feed nitrogen current, oil phase is slowly added the aqueous phase of high-speed stirred, it is in good time to continue high-speed stirred, with sodium hydroxide solution regulator solution pH value to prescribed limit; Under nitrogen current, make colostrum high pressure homogenize under 40-70MPa (for example 55-60MPa) pressure, again low pressure homogenize under 5-25MPa (for example 12-18MPa) pressure, cooling, add the saccharide solution of the suitable quantity of water preparation of recipe quantity, again this colostrum is settled to theoretical recipe quantity, stir; Use the membrane filtration of 10 μ m then; The filtrate fill in packing container, nitrogen-filled seal, pressure sterilizing, namely.

According to the needs in the preparation, can in suitable processing step, for example after colostrum forms, can add other adjuvants, as stabilizing agent, isotonic agent etc.

The lipomul that contains vitamin K1 of the present invention suits medicinal, uses such as oral and injection.Especially, the emulsion composition of the present invention drug administration by injection that can suit uses, and breast grain diameter wherein is little, and mean diameter and has the general prescription of the medicine of meeting between 0.2～1 μ m.

The lipomul that contains vitamin K1 of the present invention can comprise approach and modes such as intramuscular injection, intravenous injection, intravenous drip by approach and the mode medication of injection, but also by oral route medication, for example oral liquid, oral drop, mouth sprays etc.For example, when the administration of human time spent, the lipomul that contains vitamin K1 of the present invention, dosage for each person every day can be the vitamin K1 of 0.1mg～0mg, preferably 1mg～40mg.Can be once-a-day or multiple dosing, also can be administered once in many days.When intravenously administrable, the present invention can be contained the direct intravenous injection of lipomul of vitamin K1, intravenous injection once or intravenous drip after also it can being diluted with suitable transfusion diluent.Described transfusion diluent comprises electrolyte solution such as normal saline, glucose solution, xylitol solution, fructose soln, woods Ge Shi T3 and glycerol etc.Especially, described transfusion diluent is commercially available 10% fat emulsion injection that is used for the nutritional supplement.In addition, when oral administration, Emulsion of the present invention is oil-in-water type because of it, and it also can be gone in being diluted in appropriate liquid again and take, and described liquid is such as but not limited to water, fruit drink etc.

Emulsion composition provided by the invention can orally use, and can also inject use.Be fit to drug packaging container oral or that injection is used numerous species is arranged.When therefore mentioning " packing container " or " drug packaging container " in the present invention, they can for example be vials, for example cillin bottle, ampoule bottle, phial, and the injecting drug use packaging vial of other type or the oral medication packaging vial of other type etc.); Also can be plastic bottle, for example plastic ampoule, plastics oral liquid bottle, and the injecting drug use packaging plastic bottle of other type or the oral medication packaging plastic bottle of other type etc. can also be the drug packaging containers of other material.

The lipomul that the present invention contains vitamin K1 has good pharmaceutics performance, meets medicinal requirements.For example, the vitamin K1 emulsion composition that preparation example 1 of the present invention prepares is tested about the standard of anaphylaxis, hemolytic and the vascular stimulation test of injection systemic administration Emulsion, muscle irritation, subcutaneous irritation test according to state food and drug administration, and the result shows: the vitamin K1 fat emulsion injection of 4mg/kg and 1mg/kg does not have allergenic effect to Cavia porcellus; Vitamin K1 fat emulsion injection 0.2mg/kg～1mg/kg does not have obvious external haemolysis and agglutination to tame rabbit erythrocyte; 1mg/ml vitamin K1 fat emulsion injection 1mg/min intravenously administrable, continuous 7 days, does not have the obvious stimulation effect to the rabbit auricular vein at every day 1 time; The vitamin K1 fat emulsion injection subcutaneous injection 1ml of 10mg/ml, every day 1 time, continuous 7 days, no obvious stimulation effect.

The specific embodiment

Following examples further specify the present invention, rather than restriction the present invention.In the example below, when using the pH regulator agent material regulated, as there is not an other explanation, be to regulate with 1M sodium hydroxide solution or 1M hydrochloric acid solution, its consumption be make material particularly the pH value of solution be adjusted to setting (for example indicated value ± 0.1 scope in) or scope, be 6.5 and the target pH value is 8.0 o'clock at the pH value of current material solution for example, earlier be adjusted to 8.0 ± 0.1 with the 1M sodium hydroxide solution, if it is excessive to regulate, available 1M hydrochloric acid solution readjustment, these operations are the general technical ability that those skilled in the art possess.Preparation process purpose for example hereinafter, and done some based on the comparability of respectively giving an example and specifically describe, those skilled in the art can therefrom summarize the method that obtains pharmaceutical compositions of the present invention fully according to existing knowledge.

In the present invention's some examples hereinafter, preparation is during the present composition, as explanation in addition, used adjuvant all be the injection level other, Pei Zhi emulsion composition both can be for having orally used thus, also can be for injecting use.In the present invention's some examples hereinafter, lower alcohol or saccharide are when they are used in combination separately, and portfolio ratio is weight ratio.

In the present invention's preparation example and the prepared emulsion composition of reference examples hereinafter, as not explanation in addition, equal parameters such as the number of times of the homogenize by adjusting dispersing emulsification machine, time, the granularity of feasible breast eventually (before the sterilization) is controlled in following scope: the particle diameter of the breast grain more than 90% is between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter is less than 10% (light scattering method, wet method) greater than the breast grain of 5 μ m.

A, preparation of compositions example part: prepare emulsion composition of the present invention

Preparation example 1: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerol 2.2g,

Mannitol 2.5g,

The pH regulator agent is an amount of, adjust pH to 6.5,

Water is an amount of, adds to 100ml.

Method for making:

(1) take by weighing vitamin K1, soybean oil, Ovum Gallus domesticus Flavus lecithin, glycerol, mannitol by recipe quantity, standby.

(2) preparation of water: the water for injection that will prepare total amount about 80% adds stainless cylinder of steel, adds the glycerol of recipe quantity while stirring.

(3) preparation of oil phase: the soybean oil of recipe quantity is added in another stainless cylinder of steel, feed nitrogen current, add vitamin K1 and the Ovum Gallus domesticus Flavus lecithin of recipe quantity, be stirred to the Ovum Gallus domesticus Flavus lecithin dissolving.

(4) preparation of colostrum: feed nitrogen current, the aqueous phase that oil phase is slowly added high-speed stirred, continue high-speed stirred after at least 5 minutes, with 1M sodium hydroxide solution regulator solution pH value setting (on the setting basis, suitably increase by 0.1 pH unit, make the pH value after the Emulsion sterilization reach the value of regulation).

(5) feed nitrogen current, colostrum is behind high pressure homogenize under the 55-60MPa pressure 4 times, 1 (making that the overwhelming majority (microscopic examination has more than 95%) breast grain granularity is between 0.2～0.8um) back cooling of 12-18MPa low pressure homogenizing, the saccharide solution that adding is prepared with suitable quantity of water, again emulsion is settled to theoretical amount, stir, use the membrane filtration of 10 μ m then.

(6) the filtrate fill is in 1ml brown glass ampoule bottle, sealing by fusing behind the inflated with nitrogen, and 118 ℃ of moist heat sterilization 25min, namely.

As a kind of medicine, after this can be further with this sterilising prods labeling behind lamp inspection, packing, after the full review of sampling is qualified namely.As medicine, can change the capacity of ampoule bottle in addition, in order to the single-dose preparations of various dose is provided.This emulsion also can be packaged in the vial for oral use.

Preparation example 2: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Propylene glycol 2.5g,

Mannitol 2.5g,

The pH regulator agent is an amount of, adjust pH to 5,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 3: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerin/propylene glycol (1/1, weight ratio) 2g,

Sorbitol 3g,

The pH regulator agent is an amount of, adjust pH to 8,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 4: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerin/propylene glycol (2/1, weight ratio) 2g,

Fructose 3g,

The pH regulator agent is an amount of, adjust pH to 6,

Water is an amount of, adds to 100ml.

Method for making: the vitamin K1 in will writing out a prescription, soybean oil, Ovum Gallus domesticus Flavus lecithin and glycerin/propylene glycol heat down and abundant mix homogeneously at 70～80 ℃, in the solution that obtains, add 20ml water for injection, stir, add fructose and make dissolving, add the injection water again and make it into 100ml, with this solution of the preliminary homogenize of homogenizer, regulate pH value in case of necessity to setting.Then, under the pressure of 1100Bar, with NS1001L type high pressure homogenizer (Italian GEA Niro Soavi company) homogenize 7 times (making that overwhelming majority breast grain granularities are between 0.2～0.8um), make and contain the lipomul that vitamin K1 is 10mg among every 1ml.This lipomul can be potted in the little ampoule bottle, can adorn 1ml, 2ml, 5ml, 10ml, 20ml etc. for every, through 115 ℃ of sterilizations 30 minutes, namely gets the lipomul injection that contains vitamin K1 again.This injection is intramuscular injection or intravenous injection directly, also can instil with 5% glucose injection dilution posterior vein, can also instil with 10% fat emulsion injection dilution posterior vein.In addition, this Emulsion also can orally use, and when expection was used for oral administration, medicine can be packaged in oral packing container for example in the oral liquid vial person of one-tenth oral liquid plastic bottle.

Preparation example 5: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.5g,

Soybean oil 15g,

Soybean phospholipid 2g,

Glycerol 2g,

Mannitol/fructose (1/1) 4g,

The pH regulator agent is an amount of, adjust pH to 7,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 6: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 2.0g,

Soybean oil 7.5g,

Ovum Gallus domesticus Flavus lecithin 0.8g,

Propylene glycol 3g,

Mannitol/sorbitol (1/2) 3g,

The pH regulator agent is an amount of, adjust pH to 5.5,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 7: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 2g,

Soybean oil 15g,

Soybean lecithin 1.0g,

Glycerol 1g,

Sorbitol 1g,

The pH regulator agent is an amount of, adjust pH to 7.5,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 8: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.5g,

Soybean oil 5g,

Soybean lecithin 0.5g,

Propylene glycol 5g,

Fructose 5g,

The pH regulator agent is an amount of, adjust pH to 6,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 9: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 2g,

Glycerol 2g,

Mannitol/sorbitol/fructose (1/1/2) 4g,

The pH regulator agent is an amount of, adjust pH to 6,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 10: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.75g,

Soybean oil 8g,

Ovum Gallus domesticus Flavus lecithin 0.75g,

Glycerol 2g,

Oleic acid 0.25g,

Fructose 5g,

The pH regulator agent is an amount of, adjust pH to 7,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 11: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.5g,

Soybean oil 12g,

Ovum Gallus domesticus Flavus lecithin 1.5g,

Glycerol 4g,

Miglyol 812 2.0g

Sorbitol 3g,

The pH regulator agent is an amount of, adjust pH to 6.5,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out.

Preparation example 12: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerin/propylene glycol (1/1) 2.2g,

Mannitol/fructose (1/1) 2.2g,

Sucralose 0.001g,

The pH regulator agent is an amount of, adjust pH to 5,

Water is an amount of, adds to 100ml.

Method for making: the method with reference to record in the preparation example 1 is carried out, and sucralose and saccharide make an addition in the continuous phase.More than in each preparation example, each batch Emulsion before sterilization, the particle diameter of 94-96% breast grain between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter is less than 6% greater than the breast grain of 5 μ m; Each batch Emulsion is in sterilization and after 50 ℃ are placed 21 days, the particle diameter of 93-96% breast grain between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter all is less than 8% greater than the breast grain of 5 μ m; For example for preparation example 1,3,12 Emulsion, they are after sterilization, the particle diameter of 95%, 94% and 95% breast grain is arranged respectively between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter is respectively 8%, 6% and 7% greater than the breast grain of 5 μ m.As seen Emulsion breast grain change of granularity before and after sterilization and heat treatment is less.

The emulsion composition for preparing following reference examples, they are as the contrast reagent of emulsion composition of the present invention:

Reference examples 1: except mannitol being replaced with glucose, prescription and method for making are with preparation example 1;

Reference examples 2: except not using propylene glycol, prescription and method for making are with preparation example 2;

Reference examples 3: except not using sorbitol, prescription and method for making are with preparation example 3;

Reference examples 4: except fructose being replaced with glucose, prescription and method for making are with preparation example 4;

Reference examples 5: except not using glycerol, prescription and method for making are with preparation example 5;

Reference examples 6: except saccharide being replaced with glucose, prescription and method for making are with preparation example 6;

Reference examples 7: except not using mannitol, VK1 and glycerol consumption being replaced with respectively 3g and the 8g, prescription and method for making are with preparation example 7;

Reference examples 8: except VK1, soybean oil and propylene glycol consumption being replaced with 0.2g, 2g and 0.5g respectively, prescription and method for making are with preparation example 8;

Reference examples 9: except soybean oil, Ovum Gallus domesticus Flavus lecithin and saccharide consumption being replaced with 18g, 3g and 8g respectively, prescription and method for making are with preparation example 9;

Reference examples 10: except soybean oil, Ovum Gallus domesticus Flavus lecithin and saccharide consumption being replaced with 3g, 0.25g and 0.5g respectively, prescription and method for making are with preparation example 10;

Reference examples 11: except glycerol being replaced with Macrogol 200, prescription and method for making are with preparation example 11;

Reference examples 12: except saccharide being replaced with lactose, prescription and method for making are with preparation example 12.

Reference examples 13: except VK1 being replaced with VK2, prescription and method for making are with preparation example 1.

Reference examples 14: except VK1 being replaced with VK2, prescription and method for making are with preparation example 3.

Reference examples 15: get 1g vitamin K1, alpha-tocopherol 0.1g, safflower oil 10g, soybean lecithin 2.0g in nitrogen current, in 65 ℃ hot water, heating makes it be mixed into oil phase uniformly.About 80ml dissolving makes into water with water for injection with 1.0g poloxamer F68, glycerol 2.2g in addition.Then handle according to preparation example 1 step (4), (5) and (6), get whole emulsion.

Reference examples 16: get 1g vitamin K1, soybean oil 2g and lecithin 1g, in nitrogen current, in 65 ℃ hot water, heating makes it be mixed into oil phase uniformly.In addition the 5g glucose is dissolved in the distilled water for injection of 80ml and becomes water.Water is added in the oil phase while stirring, be placed on then in the ice while cooling off with ultrasound mulser emulsifying 20 minutes.The pH value of gained emulsion is adjusted to 6.5 with the sodium hydroxide solution of 0.1N, adds water for injection afterwards and make into 100ml.Then handle according to preparation example 1 step (5) and (6), get whole emulsion.More than in each reference examples, each batch Emulsion before sterilization, the particle diameter of 95-97% breast grain between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter all is less than 6% greater than the breast grain of 5 μ m.Each batch Emulsion is in sterilization and after 50 ℃ are placed 21 days, the particle diameter of 82-88% breast grain between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter reaches 14-23% greater than the breast grain of 5 μ m; For example for reference examples 1,3,12,16 Emulsion, they are after sterilization, the particle diameter of 83%, 84%, 87% and 85% breast grain is arranged respectively between 0.1～2.0 μ m, and particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter reaches 18%, 16%, 17% and 20% respectively greater than the breast grain of 5 μ m.As seen Emulsion breast grain change of granularity before and after sterilization and heat treatment is bigger.

B, detection method part: the method that detects emulsion composition of the present invention

Following detection side's rule provides the method that can be used for detecting emulsion composition of the present invention, these methods can also be used for measuring the treated for example internal performance after high-temperature process of emulsion composition of the present invention, can also measure some other emulsion composition for comparative test with reference to the inventive method preparation.

Detection side's rule 1: the pH value that detects emulsion composition of the present invention

Method: according to the method that two appendix VI of Chinese Pharmacopoeia version in 2010 H puts down in writing, directly measure emulsion composition of the present invention or other various samples that the present invention obtains.In the present invention's test, when providing pH value, be the average of 3 parallel assays.The pH value of some emulsion compositions of the present invention's preparation is respectively referring to the value described in preparation example 1～12 prescription (and the difference between the prescription sign value is less than 0.1 pH unit).

Detection side's rule 2: detect granularity and the distribution of emulsion composition of the present invention

Method: measure according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two appendix IX E granularity and the particle size distribution method three therapeutic methods of traditional Chinese medicine (light scattering method), adopt the laser light scattering particles distribution instrument to carry out, record and statistics are no less than the particle diameter of 500 breast grain particles.Calculate the percent (can represent with " percentile 1 " in this article) that the newborn grain number of particle diameter in certain setting scope accounts for the total statistical number of breast grain.Calculate simultaneously in the breast grain sum in particle diameter setting scope, particle diameter is greater than the percent (can represent with " percentile 2 " in this article) of the breast grain in certain setting scope.

Detection side's rule 3: the content that detects vitamin K1 in the emulsion composition of the present invention

Method:

(1) measures according to high performance liquid chromatography (2010 editions two appendix V D of Chinese Pharmacopoeia);

(2) chromatographic condition and system suitability test: being filler with octadecylsilane chemically bonded silica, is mobile phase with alcohol-water (95: 5), and column temperature is 35 ℃, and the detection wavelength is 254nm, and theoretical cam curve is calculated with vitamin K1 should be not less than 1500;

(3) preparation of need testing solution: it is an amount of that precision is measured emulsion composition, adds the isopropyl alcohol dissolving and quantitatively make to contain the solution that vitamin K1 is 0.05mg among every 1ml, as need testing solution;

(4) preparation of reference substance solution: it is an amount of to take by weighing the vitamin K1 reference substance, the accurate title, decide, adding 10% fat emulsion injection (prepares with reference to preparation example 1 of the present invention, difference is vitaminize K1 not) 1ml, add the isopropyl alcohol dissolving and quantitatively make and contain the solution that vitamin K1 is 0.05mg, product solution in contrast among every 1ml;

(5) algoscopy: get each 20 μ l of need testing solution and reference substance solution respectively, inject chromatograph of liquid, the record chromatogram is by the content of external standard method with vitamin K1 in the calculated by peak area test sample.

The content of vitamin K1 can represent with % (w/v) or mg/ml, perhaps represents with the percent of the labelled amount that is equivalent to fill a prescription.

Detection side's rule 4: the content that detects glycerol in the emulsion composition of the present invention

Method: precision is measured emulsion composition 2.5ml, puts in the conical flask, adds water 100ml, adds 5 of bromocresol purple indicator solutions, shakes up, if show acid, drips the 0.1mol/L sodium hydroxide solution, makes solution be bluish violet; If show alkalescence, should drip the 0.5mol/L sulfuric acid solution earlier is adjusted to solution and just is yellow, drip the 0.1mol/L sodium hydroxide solution again and make solution be bluish violet, add 0.7% Potassium metaperiodate. solution (facing with newly joining) 100ml, put in 37～40 ℃ the water-bath insulation 15 minutes, and jolting constantly, add 1,2-propylene glycol 3ml, placed 5 minutes, just be bluish violet with 0.1mol/L sodium hydroxide volumetric solution titration to solution, namely.The 0.1mol/L sodium hydroxide volumetric solution of every 1ml is equivalent to the C3H8O3 of 9.210mg.

The content of glycerol can represent with % (w/v) or mg/ml, perhaps represents with the percent of the labelled amount that is equivalent to fill a prescription.

Detection side's rule 5: the content that detects triglyceride in the emulsion composition of the present invention

Method:

(1) measures according to high performance liquid chromatography (two appendix VD of Chinese Pharmacopoeia version in 2010);

(2) chromatographic condition and system suitability test: be filler with silica gel, normal hexane-isopropyl alcohol-acetic acid (98.9: 1: 0.1) is mobile phase, evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 ℃) detects, the separating degree of triolein and oleic acid should be greater than 2, and the relative standard deviation of the peak area of triglyceride should be not more than 3.0%;

(3) preparation of system suitability testing liquid: get each 10mg of triglyceride and oleic acid, to the 25ml measuring bottle, with mobile phase dissolving and be diluted to scale, shake up, namely;

(4) preparation of reference substance solution: get the about 0.35g of soybean oil reference substance, accurate claim surely, put in the 50ml measuring bottle, dissolve and be diluted to scale with appearance mixed solutions such as normal hexane and isopropyl alcohols, be stock solution; Precision is measured stock solution 3ml and 4ml, puts respectively in the 25ml measuring bottle, is diluted to scale with mobile phase, shakes up, and is reference substance solution 1 and reference substance solution 2;

(5) preparation of need testing solution: precision is measured emulsion composition 4ml, puts in the 50ml measuring bottle, with appearance mixed solutions such as normal hexane and isopropyl alcohol dissolvings and be diluted to scale, shakes up; Precision is measured 3ml, puts in the 25ml measuring bottle, is diluted to scale with mobile phase, shakes up, namely;

(6) algoscopy: precision is measured reference substance solution 1, reference substance solution 2 respectively, and each 10 μ l of need testing solution alternately inject chromatograph of liquid, get calculated by peak area through chromatographic isolation, namely.

The content of triglyceride can represent with % (w/v) or mg/ml, perhaps represents with the percent of the labelled amount that is equivalent to fill a prescription.

Detection side's rule 6: the content that detects free fatty acid in the emulsion composition of the present invention

The preparation of contrast solution: get Palmic acid 0.64g, the accurate title, decide, and puts in the 500ml measuring bottle, adds the normal heptane dissolving and be diluted to scale, shakes up.

The emulsion composition of getting preparation dilutes 10 times with 5% glucose injection.

Precision is measured diluent and each 1ml of contrast solution, puts respectively in the 20ml tool plug test tube, adds the mixed solution 5.0ml of isopropyl alcohol-normal heptane-0.5mol/L sulfuric acid solution (40: 10: 1), and jolting 1 minute was placed 10 minutes.Need testing solution pipe precision adds normal heptane and each 3ml of water, and reference substance solution pipe precision adds normal heptane 2ml and water 4ml, close plug, spin upside down 10 times, left standstill at least 15 minutes, and made layering, precision is measured upper strata liquid 3ml respectively, put in the 10ml centrifuge tube, add the Nile blue indicator solution (get Nile blue 0.04g, add water 200ml, make dissolving after, add normal heptane 100ml jolting, discard the upper strata normal heptane.Repeatable operation 4 times.Take off a layer aqueous solution 20ml, add dehydrated alcohol 180ml, mixing.This liquid is put in the brown bottle, can deposit one month under the room temperature.Under logical nitrogen condition, show lavender with sodium hydroxide volumetric solution (0.01mol/L) titration to solution.Need testing solution consumes the milliliter number of sodium hydroxide volumetric solution and the ratio that reference substance solution consumes sodium hydroxide volumetric solution milliliter number, multiply by 5.0mmol/L again, amounts to into the concentration sign of free acid.

C, test routine part

Test example 1: investigates Emulsion of the present invention and make up bioavailability when orally using

Reagent: preparation example 1,2,3, each emulsion composition of 12; Reference examples 1,2,3, each emulsion composition of 12; Commercially available vitamin K1 sheet (5mg/ sheet).

Animal: rat, body weight 200-250g, male and female dual-purpose.

Medication: preparation example 1,2,3, each emulsion composition oral administration of 12, preparation example 1,2,3,12 the quiet notes administration of each emulsion composition, reference examples 1,2,3, each emulsion composition oral administration of 12, totally 12 groups of experimental animals, 6 every group.

Dosage: every kg the weight of animals gives the emulsion composition of 2mg active component.

Method: 15min, 30min, 1h, 2h, 3h, 4h, 5h, 6h, 7h, 8h, 10h, 12h, 15h, 18h, 24h, 30h, 36h, 42h, 48h blood sample collection respectively before the administration and after the administration, document with reference to Dai Li etc. (is worn jasmine, Deng, the human body pharmacokinetics of vitamin K1 soft capsule and bioequivalence Journal of Sex Research, pharmacy with face fiber crops research, 2007,15 (5): 380) method of record is handled sample, detect the line data of going forward side by side statistics.

Calculate every treated animal area (AUC) under the plasma concentration curve of taking medicine in back 48 hours respectively, calculate the percent of each oral group AUC during with respect to its corresponding prescription injection then respectively, that is:

In following formula I and formula II, preparation example n represents to number the emulsion composition of n preparation example, for example preparation example 1,2,3 or 12.In following formula II, reference examples n represents to number the emulsion composition of n reference examples, for example reference examples 1,2,3 or 12.And in formula II, reference examples numbering n is consistent with preparation example numbering n, and when namely for example calculating the oral percentage AUC of reference examples 1, it is reference examples 1 oral AUC injects AUC institute value divided by preparation example 1 percent.The result shows, preparation example 1,2,3 and 12 oral percentage AUC (%) are between 88-93%, and for example the oral percentage AUC (%) of preparation example 1 is 92%.Reference examples 1,2,3 and 12 oral percentage AUC (%) are between 76-84%, and for example the oral percentage AUC (%) of reference examples 1 is 83%.The oral percentage AUC (%) of commercially available vitamin K1 tablet is 73%.

Test example 2: the pharmaceutical property of emulsion composition of the present invention is investigated

1, The content of vitamin K1:Measure according to method shown in above, account for the percent that prescription indicates content in the content of test.The result shows that VK1 content (%) is all in 98.8%～101.3% scope in each sample of preparation example 1-12 above.

2, glycerol content:Measure according to method shown in above.The result shows that VK1 content (%) is all in 97.5%～102.7% scope in each sample of preparation example 1-12 above.

3, triglyceride content:Measure according to method shown in above.Account for the percent of soybean oil sign content in the prescription in the content of test.The result shows that VK1 content (%) is all in 97.1%～104.3% scope in each sample of preparation example 1-12 above.

Claims (10)

1. emulsion composition, it comprises:

Wherein, described lower alcohol is selected from propylene glycol, glycerol or its combination, and described saccharide is selected from sorbitol, mannitol, fructose, glucose or its combination.

2. according to the emulsion composition of claim 1, it is measured according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two appendix IX E granularity and the particle size distribution method three therapeutic methods of traditional Chinese medicine light scattering method, and the particle diameter of the breast grain more than 90% is between 0.01～5.0 μ m.

3. according to the emulsion composition of claim 2, wherein particle diameter greater than with the breast grain sum that equals 1 μ m in, particle diameter is less than 20% greater than the breast grain of 5 μ m.

4. according to the emulsion composition of claim 1, wherein also comprise correctives, antiseptic and/or pH regulator agent.

5. according to the emulsion composition of claim 1, its pH value is pH5-8.

6. according to the emulsion composition of claim 1, it comprises:

The vitamin K1 of 0.9-1.1%;

9～11% soybean oil;

1.0～1.5% phospholipid;

2～4% lower alcohol;

1～5% saccharide.

7. according to the emulsion composition of claim 1, it comprises:

8. according to each emulsion composition of claim 1-7, it is oil-in-water emulsion.

9. emulsion composition according to Claim 8, it is Emulsion for oral use, or injectable emulsion.

10. prepare the method for each emulsion composition of claim 1-9, it may further comprise the steps:

(1) take by weighing vitamin K1, soybean oil, phospholipid, lower alcohol, saccharide by recipe quantity, and other optional material, standby;

(2) will prepare in the water adding Agitation Tank of total amount 60-90%, add the lower alcohol of recipe quantity, stir, make water;

(3) soybean oil with recipe quantity adds in another Agitation Tank, feeds nitrogen current, adds vitamin K1 and the phospholipid of recipe quantity, is stirred to each material dissolution, makes oil phase;

(4) feed nitrogen current, oil phase is slowly added the aqueous phase of high-speed stirred, it is in good time to continue high-speed stirred, regulates its pH value to prescribed limit with sodium hydroxide solution, gets colostrum;

(5) under nitrogen current, make colostrum high pressure homogenize under 40-70MPa pressure, low pressure homogenize under 5-25MPa pressure is cooled off again;

(6) add saccharide with an amount of water-soluble recipe quantity, add water again and be settled to theoretical recipe quantity, stir, in case of necessity with sodium hydroxide or hydrochloric acid solution regulator solution pH value to prescribed limit, use the membrane filtration of 10 μ m then;

(7) the filtrate fill is in the drug packaging container, and sealing is randomly carried out pressure sterilizing, namely.