CN102552135A

CN102552135A - Method for preparing vitamin K1 emulsion, and vitamin K1 emulsion

Info

Publication number: CN102552135A
Application number: CN2012100300524A
Authority: CN
Inventors: 傅卫国; 梅丹; 黄伟静; 王佩贤
Original assignee: XI'AN ANJIAN PHARMACEUTICAL CO Ltd
Current assignee: Guangzhou Anjian Industrial Development Co., Ltd.
Priority date: 2012-02-13
Filing date: 2012-02-13
Publication date: 2012-07-11

Abstract

The invention relates to a method for preparing a vitamin K1 emulsion, and a vitamin K1 emulsion. The emulsion provided by the invention comprises components of: 0.5-2% of vitamin K1, 7.5-15% of soybean oil, 0.8-2% of phosphatide, 1.5-3% of glycerin, and balance of water. The pH value of the emulsion is 3.5-7.0. The method provided by the invention comprises a step for regulating the pH value of the emulsion composition to 3.5-7.0. The vitamin K1 emulsion provided by the invention has good medicine properties.

Description

The method and the vitamin K1 Emulsion that prepare vitamin K1 Emulsion

Technical field

The invention belongs to medical technical field, relate to a kind of method for preparing of Emulsion, particularly a kind of method for preparing vitamin K1 Emulsion.The invention still further relates to the vitamin K1 Emulsion for preparing by the inventive method.

Background technology

Vitamin K1 is the clarifying thick liquid of a kind of Yellow-to-orange, and odorless or odorless are almost met light and be prone to decompose.Vitamin K1 is prone to dissolve in chloroform, ether or vegetable oil, dissolves in the ethanol part omitted, and is 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 from alfalfa, purifying the earliest, and the called after vitamin K1, and people just utilize vitamin K1 to do hemorrhage very early.Its hemostatic medical value 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 in food, adds certain feedstuff; Found to be rich in this feedstuff a kind of material of hemostatic immediately afterwards, this hemostatic material is exactly a 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 comes to light 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 a clinical commonly used drug.

Vitamin K1 is a 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 for many years basic research and clinical use, 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, is clinical commonly used drug at present.

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 a main component of forming prothrombing in the blood of human body, can promote liver to make thrombinogen, organizes when hemorrhage; Platelet is destroyed; Discharge thrombokinase and calcium ion, thrombinogen can combine 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 contraction of smooth muscle spasm property, removes oxygen-derived free radicals, and has parahormone appearance 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, helps 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 through lax spasm stops the internal organs pain due to the multiple illness, like biliary colic, renal colic, and ureter and cystolith angor etc.; Effective percentage can reach 80～90%, and analgesic effect is fast, persistent, safety, curative effect are high.

The shortage of vitamin K1 can cause blood not solidify, and is mostly neonate because of vitamin K1 lacks hemorrhage patient in clinical.1～2 day diet of neonate birth is limited, and the intake of vitamin K is few, and vitamin content is low in the breast milk; Aseptic in the intestinal in addition, influence 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 a hemorrhagic disease of the newborn.The 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 in 2 months, occur in first week after the birth, and the both of these case morbidity is anxious, be prone to cause the intracranial hemorrhage damage, disability rate height, mortality rate are also high.The sickness rate of China is up to 0.5～2%, and the geographic sickness rate in Yantai is up to 9.03%, and China is the hotspot of vitamin K deficiency property 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 of 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 is meant by various different pathogens and other caused by factors pneumonias, causes the lung microcirculation disturbance, 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 contraction of smooth muscle spasm property, removes oxygen-derived free radicals; And have parahormone appearance 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; The also expansible blood vessel of vitamin K1 improves blood fortune, alleviates heart front and back load, corrects heart failure; Excited respiratory center suppresses cerebral cortex, improves brain microcirculation and prevents cerebral edema; Can reduce airway resistance, increase alveolar ventilation, the ventilation ventilation of alleviating crisis life is depleted; Help exhaling the correction that declines, so vitamin K1 can alleviate and improve the clinical symptoms of infantile pneumonia, also can reduce the case fatality rate of severe pneumonia simultaneously greatly; Shorten the hospital stays, measure nonresponders such as aminophylline, promethazine, dexamethasone, oxygen uptake are used again, still can receive promising result; Someone assists a ruler in governing a country severe pneumonia of infants 100 examples with vitamin K1 and treats 50 examples, obtains effect preferably.

Vitamin K1 treatment bronchiolitis and bronchial asthma report are also many, the domestic 41 routine children asthmatic brouchitis case intravenous applications vitamin K1 treatment asthmatic bronchitiss that research is arranged from institute in October in October, 1998 to calendar year 2001, satisfactory effect.Behind the child vein instillation vitamin K1, infant is quiet rapidly, and sleeping very soon, 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, through the smooth muscle of lax spasm, and stops the internal organs pain due to the multiple illness; Like 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 property 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.Pettifor in 1975 etc. find that the pregnant woman oral anticoagulant produces baby's nasal bone hypoplasia, 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 are positive prevention and therapeutical effect; 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, and this albuminoid is called vitamin K dependent protein matter again, and the γ-Qiang Jiguansuan residue after its activation has combination Ca ⁺⁺Characteristic, thereby make its biological activity of vitamin k-dependent protein performance, work as vitamin K deficiency, this proteinoid hydroxylation obstacle, therefore its biological activity also reduces.Vitamin K and osteoporotic relation have been explained in more bases and clinical trial.Other has the recycle of vitamin K in the anticoagulant body capable of blocking to cause the clear proof of vitamin K deficiency.The side effect of clinical report anticoagulant to bone metabolism arranged in 1975, and 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 the treatment or the prevention of liver inflammatory diseases.Research confirms 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, quickens 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, needs antifreeze preservation, if there is oil droplet to separate 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 PhytonadioneInjectable Emulsion of the alternative American Pharmacopeia of this micro emulsion composition; USP; Can abbreviate PIE-USP as, its former name is a vitamin K 1 injection) be used 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 strong effect surfactant.Although nomenclature of drug is recited as the injection breast; But vitamin K1 injectable emulsion (USP) former grind product be Merck & Co., Inc.

its in fact do not contain any oil, therefore be 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 through 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 more a large amount of 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.Than the safety of oil for injection (for example safflower oil, the Fructus Canarii albi wet goods) soybean oil of other kind and effectiveness is what fully to have proved, and the heavy dose of soybean oil that provides with the fat emulsion injection form has used many decades clinically.

People still expect to have new vitamin K 1 injection for example the vitamin K1 fat emulsion injection be used for clinical.

Summary of the invention

The objective of the invention is to be the clinical for example vitamin K1 fat emulsion injection of a kind of new vitamin K 1 injection that provides, this injection has the beneficial effect that at least a for example the present invention puts down in writing.The inventor finds, by the vitamin K 1 injection that the special formulation that comprises about 10% soybean oil prepares, be the fat milk that a kind of performance can satisfy the clinical application demand fully, and it has advantage uniquely in preparation technology.The present invention is based on this discovery and be accomplished.

For this reason, first aspect present invention provides the method for preparing emulsion composition, and said emulsion composition comprises:

Vitamin K1 0.5-2%,

Soybean oil 7.5～15%,

Phosphatidase 10 .8～2%,

Glycerol 1.5～3%,

Water is an amount of, adds to 100%;

And the pH value of said emulsion composition is 3.5～7.0;

This method may further comprise the steps:

(1) take by weighing vitamin K1, soybean oil, phospholipid, glycerol by recipe quantity, subsequent use;

(2) water for injection that will prepare the about 60-90% of total amount (for example about 80%) adds stainless cylinder of steel, adds the glycerol of recipe quantity, stirs, and makes water;

(3) soybean oil with recipe quantity adds in another stainless cylinder of steel, feeds nitrogen current, adds the 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, colostrum;

(5) with pH value to 3.8～6.8 of this colostric fluid of sodium hydroxide solution adjusting, add the injection 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 3.8～6.8;

(6) 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, uses the membrane filtration of 10 μ m then;

(7) the filtrating fill is filled the nitrogen sealing by fusing in ampoule bottle, and pressure sterilizing promptly gets.

According to the method for first aspect present invention, it is included in the suitable processing step, adds other 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 first 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 first aspect present invention, before compositions encapsulation and sterilization, measure with or the pH value of regulating said composition be very easy; And (promptly becoming) should not be regulated the pH value of said composition usually again after compositions encapsulation and sterilization.It will be apparent to those skilled in the art that; 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,, be meant sterilization treatment pH value afterwards if specify; For example in the preparation example hereinafter 1; When describing composite formula, mention " the pH regulator agent is an amount of, adjust pH to 6.0 " be meant in prescription or the dosing process in to add an amount of pH regulator agent an amount of; Make medicinal liquid through corresponding method for preparing processing and after pressure sterilizing obtains final emulsion composition; The pH value of this emulsion composition is about 6.0, and for example pH value is in 6.0 ± 0.2 scope, and for example pH value is in 6.0 ± 0.1 scope.Yet be to be understood that into, in the for example step of the said method of preceding text first aspect present invention " pH value to 3.8～6.8 of this colostric fluid are regulated in (5) with sodium hydroxide solution ", pH value wherein is meant the pH value of the colostrum that sterilized is handled.

According to the method for first aspect present invention, the pH value of wherein said emulsion composition is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.

According to the method for first aspect present invention, the method that wherein said emulsion composition is pressed under two appendix VI of Chinese Pharmacopoeia version in 2010 H item is measured, and pH value is 3.5～7.0.In one embodiment, pH value is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.

According to the method for first aspect present invention, also comprise an amount of pH regulator agent in the wherein said emulsion composition, and the pH value of regulating this emulsion composition is 3.5～7.0.In one embodiment, pH value is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.In one embodiment, said pH regulator agent is sodium hydroxide and/or hydrochloric acid.

According to the method for first aspect present invention, wherein said emulsion composition comprises:

Vitamin K1 0.75-1.5%,

Soybean oil 8～12%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.75～2.5%,

Water is an amount of, adds to 100%.

Vitamin K1 0.9-1.1%,

Soybean oil 9～11%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.95～2.45%,

Water is an amount of, adds to 100%.

Vitamin K1 1.0%,

Soybean oil 10%,

Phosphatidase 11 .2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%.

Method according to first aspect present invention; Wherein said emulsion composition is measured according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two the 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 70% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.1～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.1～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.1～1.0 μ m.In one embodiment, particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is less than 20% greater than the breast grain of 1 μ m, preferably is less than 15%, is less than 10%, is less than 5%.

Method according to first aspect present invention; Wherein said emulsion composition is according to 2010 editions two said HPLies 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: use octadecylsilane chemically bonded silica to be filler; With 95: 5 alcohol-water mixed liquor of volume ratio is mobile phase; 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 said compositions, adds the isopropyl alcohol dissolving and quantitatively process 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; The accurate title, decide; (in one embodiment, this 10% fat emulsion injection is 10% fat emulsion injection of vitaminize K1 preparation according to the method for hereinafter embodiment 1 of the present invention but not to add 10% fat emulsion injection; In one embodiment, this 10% fat emulsion injection is 10% a commercially available fat emulsion injection) 1ml, add isopropyl alcohol dissolving and quantitatively process and contain the solution that vitamin K1 is about 0.05mg among every 1ml, as reference substance solution;

(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 said compositions of calculated by peak area.

According to the method for first aspect present invention, wherein said emulsion composition 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 said 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 and be adjusted to solution and just be yellow, drip the 0.1mol/L sodium hydroxide solution again and make solution be bluish violet; Add and face, put in 37～40 ℃ the water-bath insulation 15 minutes with the 0.7% Potassium metaperiodate. solution 100ml that newly joins, and jolting constantly; Add 1; 2-propylene glycol 3ml placed 5 minutes, just was bluish violet with 0.1mol/L sodium hydroxide volumetric solution titration to solution; Promptly 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.

Method according to first aspect present invention; Wherein said emulsion composition is according to 2010 editions two said HPLies 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: use silica gel to be filler; With volume ratio is that normal hexane-isopropyl alcohol-acetic acid mixed liquor of 98.9: 1: 0.1 is a mobile phase; Evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 ℃) detects; Triolein and oleic separating degree 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, promptly get;

(3) preparation of reference substance solution: get the about 0.35g of soybean oil reference substance, the accurate title, decide, and puts 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 said 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, and promptly gets;

(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 method for first aspect present invention, wherein said emulsion composition 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 method for first aspect present invention, wherein said emulsion composition 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 method for first aspect present invention, wherein said emulsion composition comprises 0.8～2% phospholipid, preferably comprises 1.0～1.5% phospholipid, preferably comprises about 1.2% phospholipid.

According to the method for first aspect present invention, wherein said emulsion composition comprises 1.5～3% glycerol, preferably comprises 1.75～2.5% glycerol, preferably comprises 1.95～2.45% glycerol, preferably comprises about 2.2% glycerol.

According to the method for first aspect present invention, wherein said emulsion composition is the oil-in-water type injectable emulsion.

Second aspect present invention provides emulsion composition, and it comprises:

Vitamin K1 0.5-2%,

Soybean oil 7.5～15%,

Phosphatidase 10 .8～2%,

Glycerol 1.5～3%,

Water is an amount of, adds to 100%.

According to the emulsion composition of second aspect present invention, its pH value is 3.5～7.0.In one embodiment, pH value is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.

According to the emulsion composition of second aspect present invention, its method of pressing under two appendix VI of Chinese Pharmacopoeia version in 2010 H item is measured, and pH value is 3.5～7.0.In one embodiment, pH value is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.

According to the emulsion composition of second aspect present invention, wherein also comprise an amount of pH regulator agent, and the pH value of regulating this emulsion composition is 3.5～7.0.In one embodiment, pH value is 4.0～7.0.In one embodiment, pH value is 4.0～6.5.In one embodiment, said pH regulator agent is sodium hydroxide and/or hydrochloric acid.

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

Vitamin K1 0.75-1.5%,

Soybean oil 8～12%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.75～2.5%,

Water is an amount of, adds to 100%.

Vitamin K1 0.9-1.1%,

Soybean oil 9～11%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.95～2.45%,

Water is an amount of, adds to 100%.

Vitamin K1 1.0%,

Soybean oil 10%,

Phosphatidase 11 .2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%.

According to the emulsion composition of second aspect present invention, it is measured according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two the 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 70% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is below 1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.01～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.05～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 70% is between 0.1～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 80% is between 0.1～1.0 μ m.In one embodiment, the particle diameter of the breast grain more than 90% is between 0.1～1.0 μ m.In one embodiment, particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is less than 20% greater than the breast grain of 1 μ m, preferably is less than 15%, is less than 10%, is less than 5%.

Emulsion composition according to second aspect present invention; It is according to 2010 editions two said HPLies 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):

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):

Emulsion composition according to second aspect present invention; It is according to 2010 editions two said HPLies 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):

According to the emulsion composition of second 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 second 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 second 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 second aspect present invention, it comprises 1.5～3% glycerol, preferably comprises 1.75～2.5% glycerol, preferably comprises 1.95～2.45% glycerol, preferably comprises about 2.2% glycerol.

According to the emulsion composition of second aspect present invention, it is the oil-in-water type injectable emulsion.

The documents and materials of the present invention's citation, its full content is incorporated this paper by reference into.Of the present invention arbitrary aspect, any one or more technical characterictics of its arbitrary embodiment 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,, are being as the criterion with the present invention when inconsistent with this general sense.

In the present invention, for writing conveniently, vitamin K1 can simply be written as VK1.Those skilled in the art know, and Menaquinone K6 (can be abbreviated as VK2) is a kind of at aspect materials similar with VK1 such as structure, physicochemical property and BAs.

The present invention provides a kind of fluid composition generally, and main carrier wherein is a water, and solute comprises liquid substance (for example glycerol) and/or solid matter (for example be semi-solid Ovum Gallus domesticus Flavus lecithin).Therefore, for simplicity, in the present invention, when under the situation of indicated concentration percent, using " % ",, be meant weight/volume percent like not explanation in addition; For example.For the prescription that contains " vitamin K1 is 1%, soybean oil 10%, Ovum Gallus domesticus Flavus lecithin 1.2%, glycerol 2.2%, water add to 100% in right amount ", it is meant that containing vitamin K1 in every 100ml prescription is 1g, soybean oil 10g, Ovum Gallus domesticus Flavus lecithin 1.2g, glycerol 2.2g 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 1.0 μ m " can be meant the particle diameter of measuring the breast grain particle of some according to (for example microscopic method, light scattering method etc.) someway; For example measure about 1000 breast grain particles; Then particle diameter is dropped on population in the following scope of 1.0 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 0.5 μ m ", like not explanation in addition, the particle diameter that is meant the breast grain is between 0.2 μ m～1.0 μ m.

In the present invention, term " 10% fat emulsion injection ", like explanation in addition, be meant according to method of preparing emulsion, basically can intravenous Emulsion by what comprise that following formulation obtains:

Soybean oil 10%,

Ovum Gallus domesticus Flavus lecithin 1.2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%.

The present invention is the emulsion composition of injection, and 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 glycerol is normally brought into play the effect of osmotic pressure regulator; Further, water for injection uses as the water solvent.

Undoubtedly, the adjuvant that the present invention is used comprises soybean oil, is to meet necessarily requiredly, and for example, during fat emulsion for injection, these adjuvants should use for injection in preparation.This is well known to a person skilled in the art.

Phospholipid of the present invention is to be selected from following purified phospholipid: Ovum Gallus domesticus Flavus lecithin, soybean lecithin, cephalin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline etc.They can be bought from market, and also available usual method prepares like 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, like coemulsifier, stabilizing agent, osmotic pressure regulator, pH value regulator etc.

As coemulsifier can be that the carbon number that in every 1000ml lipomul, contains 50g or following amount is 6～22; Preferred carbon number is 12～20 fatty acid or its physiologically acceptable salt; This fatty acid can be a straight chain, also can be side chain, preferably the 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, like sodium salt, potassium salt, calcium salt etc.

As coemulsifier can also be the medium chain fatty glyceride that in every 1000ml lipomul, contains 50g or following amount, and said medium chain fatty glyceride is 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 sad triglyceride with capric acid of satisfied fatty acid, or its commercial product is like Miglyol812 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 processing is 0.5～3, and preferred osmotic pressure ratio is 0.8～1.5.

In the present invention, mention " Chinese Pharmacopoeia ", be meant Pharmacopoeia of People's Republic of China, dated especially as not, be meant the version of version in 2010; As not dated especially, be meant 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, mention emulsion composition of the present invention or when 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 material wherein.For example for Emulsion of the present invention, the pH value of before branch installs in the ampoule bottle, being measured 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 is meant and sterilizes.Therefore in production practices, when before sterilization, preparing burden, can the possible variation of with due regard to this sterilization front and back pH value; If for example pH value can reduce by 0.1 unit after sterilization, and the target pH value is 6.0, the pH value of the emulsion composition of then being prepared before the sterilization is adjusted to 6.1 and is advisable.

Lipomul provided by the invention can prepare by following method:

Take by weighing vitamin K1, soybean oil, phospholipid, glycerol by recipe quantity, subsequent use; The water for injection of preparation total amount about 80% is added stainless cylinder of steel, add the glycerol of recipe quantity while stirring; The soybean oil of recipe quantity is added in another stainless cylinder of steel, feed nitrogen current, add the 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, to prescribed limit, again this colostrum is settled to theoretical recipe quantity with sodium hydroxide solution regulator solution pH value, stirs; 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, uses the membrane filtration of 10 μ m then; The filtrating fill is filled the nitrogen sealing by fusing in ampoule bottle, pressure sterilizing promptly gets.

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

The lipomul that contains vitamin K1 of the present invention suits medicinal, uses with injection such as oral.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 through the approach and the mode medication of injection.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 0.1mg～50mg, preferably 1mg～40mg.Can be once-a-day or multiple dosing, also can be administered once in many days.When intravenously administrable, can the present invention be contained the direct intravenous injection of lipomul of vitamin K1, intravenous injection once or intravenous drip after also can it 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 T ₃With glycerin etc.Especially, described transfusion diluent is commercially available 10% fat emulsion injection that is used for the nutritional supplement.

The lipomul that the present invention contains vitamin K1 has good pharmaceutics performance, meets medicinal requirements.For example; The vitamin K1 emulsion composition that the embodiment of the invention 1 prepares makes an experiment according to the standard of state food and drug administration about anaphylaxis, hemolytic and the vascular stimulation test of injection systemic administration Emulsion, muscle irritation, subcutaneous irritation test, 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 is regulated, as not having other explanation, is 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 5.0 o'clock for example, be adjusted to 5.0 ± 0.1 with the 1M hydrochloric acid solution earlier at the pH value of current material solution, if regulate excessive; Available 1M sodium hydroxide solution readjustment, these operations are the general technical ability that those skilled in the art possess.Hereinafter preparation process purpose for example, 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.

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

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,

The pH regulator agent is an amount of, adjust pH to 6.0,

Water is an amount of, adds to 100ml.

Method for making:

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

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

(3) preparation of oil phase: the soybean oil (injection) of recipe quantity is added in another stainless cylinder of steel, feed nitrogen current, add the 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 (and/or 1M hydrochloric acid) regulator solution pH value setting (on the setting basis, suitably increase by 0.1 pH unit, make Emulsion sterilization pH value afterwards reach the value of regulation); Again colostrum is settled to theoretical amount, stirs.

(5) feed nitrogen current, colostrum is behind high pressure homogenize under the 55-60MPa pressure 4 times, and 1 (making that the overwhelming majority (microscopic examination has more than 95%) breast grain granularity is between 0.2～0.8um) postcooling of 12-18MPa low pressure homogenizing is used the membrane filtration of 10 μ m then.

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

As a kind of medicine, after this can be further with this sterilising prods labeling behind lamp inspection, packing promptly gets after the full inspection of sampling is qualified.As medicine, can change the capacity of ampoule bottle in addition, so that the single-dose preparations of various dose is provided.

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,

Glycerol 2.2g,

The pH regulator agent is an amount of, adjust pH to 5.0,

Water is an amount of, adds to 100ml.

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

Preparation example 3: preparation art invention emulsion composition

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerol 2.2g,

The pH regulator agent is an amount of, adjust pH to 4.0,

Water is an amount of, adds to 100ml.

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,

Glycerol 2.2g,

The pH regulator agent is an amount of, adjust pH to 7.0,

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 glycerol heat down and abundant mix homogeneously at 70～80 ℃; In the solution that obtains, add 20ml water for injection; Stir; Add the injection water and make it into 100ml,, regulate pH value in case of necessity to setting with this solution of the preliminary homogenize of homogenizer.Then, under the pressure of 1100Bar,, make and contain the lipomul that vitamin K1 is 10mg among every 1ml 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).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, promptly gets the lipomul injection that contains vitamin K1 again.This injection is intramuscular injection or intravenous injection directly, also can use 5% glucose injection dilution posterior vein to instil, and can also use 10% fat emulsion injection dilution posterior vein to instil.

Preparation example 5: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.5g,

Soybean oil 15g,

Ovum Gallus domesticus Flavus lecithin 2g,

Glycerol 3g,

The pH regulator agent is an amount of, adjust pH to 4.5,

Water is an amount of, adds to 100ml.

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,

Glycerol 1.5g,

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

Water is an amount of, adds to 100ml.

Preparation example 7: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.5g,

Soybean oil 8g,

Soybean lecithin 1.5g,

Glycerol 2.5g,

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

Water is an amount of, adds to 100ml.

Preparation example 8: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.75g,

Soybean oil 12g,

Soybean lecithin 1.0g,

Glycerol 1.75g,

The pH regulator agent is an amount of, adjust pH to 6.0,

Water is an amount of, adds to 100ml.

Preparation example 9: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.1g,

Soybean oil 9g,

Ovum Gallus domesticus Flavus lecithin 1.5g,

Glycerol 2.2g,

Propylene glycol 0.2g,

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

Water is an amount of, adds to 100ml.

Preparation example 10: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.0g,

Soybean oil 10g,

Ovum Gallus domesticus Flavus lecithin 1.2g,

Glycerol 1.95g,

Oleic acid 0.25g,

The pH regulator agent is an amount of, adjust pH to 6.2,

Water is an amount of, adds to 100ml.

Preparation example 11: 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,

Cholesterol 0.25g

The pH regulator agent is an amount of, adjust pH to 5.0,

Water is an amount of, adds to 100ml.

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,

Glycerol 2.2g,

Miglyol?812 2.5g，

The pH regulator agent is an amount of, adjust pH to 4.5,

Water is an amount of, adds to 100ml.

The emulsion composition for preparing following reference examples 1～12, they are as the contrast reagent of emulsion composition of the present invention:

Reference examples 1: except pH value is 3.5, prescription and method for making are with preparation example 1;

Reference examples 2: except pH value is 3.0, prescription and method for making are with preparation example 2;

Reference examples 3: except pH value is 7.6, prescription and method for making are with preparation example 3;

Reference examples 4: except pH value is 8.5, prescription and method for making are with preparation example 4;

Reference examples 5: except pH value is 9.0, prescription and method for making are with preparation example 5;

Reference examples 6: except pH value is 9.5, prescription and method for making are with preparation example 6;

Reference examples 7: except pH value is 7.5, prescription and method for making are with preparation example 7;

Reference examples 8: except pH value is 3.0, prescription and method for making are with preparation example 8;

Reference examples 9: except pH value is 3.5, prescription and method for making are with preparation example 9;

Reference examples 10: except pH value is 3.0, prescription and method for making are with preparation example 10;

Reference examples 11: except pH value is 8.0, prescription and method for making are with preparation example 11;

Reference examples 12: except pH value is 2.8, prescription and method for making are with preparation example 12.

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

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

Reference examples 15: except VK1 is replaced with the VK2, prescription and method for making are with preparation example 7.

Reference examples 16: except soybean oil is replaced with the olive oil, prescription and method for making are with preparation example 1.

Reference examples 17: 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.In addition the about 80ml dissolving of 1.0g poloxamer F68, glycerol 2.2g and water for injection is made into water.Then handle, get whole emulsion according to preparation routine 1 step (4), (5) and (6).

Reference examples 18: 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, get whole emulsion according to routine 1 step of preparation (5) and (6).

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 to measure the treated for example internal performance after high-temperature process of emulsion composition of the present invention, can also measure with reference to some other of the inventive method preparation to be used for the emulsion composition of comparative test.

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

Method:, directly measure emulsion composition of the present invention or other various samples that the present invention obtains according to the method that two appendix VI of Chinese Pharmacopoeia version in 2010 H is put down in writing.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 the 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 the 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 below 1.0 μ m accounts for the total statistical number of breast grain.Calculate simultaneously particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is greater than the percent of the breast grain of 1 μ m (in this article can with " percentile 2 " expression).

Generally speaking, but as the Emulsion of used for intravenous injection, the breast grain number of particle diameter below 1.0 μ m accounts for the percent of the total statistical number of breast grain, and promptly percentile 1 should be more than 70%, preferably more than 80%, preferably more than 90%.In addition, generally speaking, but as the Emulsion of used for intravenous injection, particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is greater than the percent of the breast grain of 1 μ m, promptly percentile 2 should be less than 20%, preferably is less than 15%, is less than 10%, is less than 5%.

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

Method:

(1) measures according to HPLC (2010 editions two appendix V D of Chinese Pharmacopoeia);

(2) chromatographic condition and system suitability test: using octadecylsilane chemically bonded silica to be filler, 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 process 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; (preparing example 1 with reference to the present invention prepares to add 10% fat emulsion injection; Difference is vitaminize K1 not) 1ml, add the isopropyl alcohol dissolving and quantitatively process and contain the solution that vitamin K1 is 0.05mg among every 1ml, as reference substance solution;

(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 use % (w/v) or mg/ml to represent, perhaps representes 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 and be adjusted to solution and just be yellow, drip the 0.1mol/L sodium hydroxide solution again and make solution be bluish violet; Add 0.7% Potassium metaperiodate. solution (facing) 100ml, put in 37～40 ℃ the water-bath insulation 15 minutes with newly joining, 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, promptly get.The 0.1mol/L sodium hydroxide volumetric solution of every 1ml is equivalent to the C3H8O3 of 9.210mg.

The content of glycerol can use % (w/v) or mg/ml to represent, perhaps representes 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 HPLC (two appendix VD of Chinese Pharmacopoeia version in 2010);

(2) chromatographic condition and system suitability test: use silica gel to be filler; Normal hexane-isopropyl alcohol-acetic acid (98.9: 1: 0.1) is a mobile phase; Evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 ℃) detects; Triolein and oleic separating degree 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, promptly get;

(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, be stock solution with appearance mixed solutions such as normal hexane and isopropyl alcohols; 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, and promptly gets;

(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, promptly get.

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

C, Test Example part

Test Example 1: the pharmaceutical property of emulsion composition of the present invention is investigated

1, the content of vitamin K1: measure according to method shown in the preceding text, account for the percent of prescription sign content in the content of test.VK1 content (%) result of 12 emulsion compositions of the present invention of preparation example 1 to preparation example 12 all labelled amount 97.7%～102.6% between, VK1 content (%) result of for example preparation example 1,4,5,11 is respectively 97.7%, 100.2%, 102.6%, 99.2% of labelled amount.

2, glycerol content: measure according to method shown in the preceding text.Content in test accounts for the percent that prescription indicates content.The result shows: glycerol content (%) result of 12 emulsion compositions of the present invention of preparation example 1 to preparation example 12 all labelled amount 94.6%～105.1% between.

3, triglyceride content: measure according to method shown in the preceding text.Account for the percent of soybean oil sign content in the prescription in the content of test.The result shows: triglyceride content (%) result of 12 emulsion compositions of the present invention of preparation example 1 to preparation example 12 all the soybean oil labelled amount 93.9%～106.1% between.

In addition, be index with above-mentioned three parameters and pH value, a breast grain granularity, investigate 12 emulsion compositions of the present invention shading under about 25 ℃ room temperature of preparation example 1 to preparation example 12 and preserved 24 months.The result shows:

The pH value of (1) 12 sample is compared with its initial value respectively, changes all less than 0.3 pH value unit, and all in the acceptable scope of the human body of pH value 3.5～7.

The breast grain granularity of (2) 12 samples:

In the time of 0 month, the breast grain particle diameter more than 85% is about 0.5 μ m; Greater than with the breast grain sum that equals 0.5 μ m in, be less than 6% greater than the breast grain of 1 μ m, do not detect breast grain greater than 5 μ m;

In the time of 24 months, the breast grain particle diameter more than 85% is about 0.5 μ m; Greater than with the breast grain sum that equals 0.5 μ m in, be less than 6% greater than the breast grain of 1 μ m, do not detect breast grain greater than 5 μ m.

The vitamin K1 content of (3) 12 samples is compared with its initial value respectively, all initial value 94.2%～103.7% between, for example preparation example 1 is 96.3%.

The glycerol content of (4) 12 samples is compared with its initial value respectively, all initial value 93.5%～107.8% between, for example preparation example 4 is 98.6%.

The triglyceride content of (5) 12 samples is compared with its initial value respectively, all initial value 94.5%～108.4% between, for example preparation example 6 is 101.2%.

Visible by above result, vitamin K1 lipomul of the present invention has good pharmaceutical property.

Test Example 2: investigate the influence of Semen sojae atricolor oil mass to filling a prescription in the vitamin K1 prescription

The emulsion composition that contains vitamin K1 with 6 different formulations preparations; Its prescription and method for making are referring to preparation example 1; The amount of the soybean oil that difference is to use; The amount of the soybean oil that in 6 prescriptions, uses is respectively 0%, 2.5%, 5%, 7.5%, 10%, 15% different samples, and it is newborn with eventually in preparation example 1 said step (4) and step (6), to obtain colostrum respectively.The colostrum of difference prescription put to leave standstill in the color comparison tube observe the layering situation after 6 hours.Whole breast is put in the centrifuge tube to observe the layering situation after 10000rpm * 10min centrifugal treating, simultaneously according to the method mensuration of " detection side's rule 2 " newborn eventually percentile 1 (" percent 1 ") and percentile 2 (" percent 2 ").The result sees the following form:

It is thus clear that stable emulsion is useful to the amount of about 7-15% soybean oil that use the present invention advocates for forming more.

Test Example 3: measure emulsion composition in sterilization forward and backward granularity and changes in distribution

Emulsion composition to each preparation example and reference examples preparation in the preceding text " A, preparation of compositions example part "; Method according to detection side's rule 2; Measure whole Emulsion that these examples make (118 ℃ of moist heat sterilizations; 25min) the variation of the granularity of forward and backward breast grain and distribution aspect (that is, for example, step (6) through filling the Emulsion of nitrogen sealing by fusing gained in the preparation routine 1; They are in the variation of forward and backward breast grain particle diameter of moist heat sterilization and distribution), respectively with percentile 1 (%) and percentile 2 (%) expression.Result such as following table.

In last each sample of table, all do not detect the breast grain of particle diameter before the sterilization greater than 5um; Each prepares routine sample and after sterilization, does not also detect the breast grain of particle diameter greater than 5um; The sample of indivedual reference examples detects the breast grain of particle diameter greater than 5um, for example reference examples 3,4,7,11,15-17 etc. after sterilization.

Claims

1. the method for preparing emulsion composition, said emulsion composition comprises:

Vitamin K1 0.5-2%,

Soybean oil 7.5～15%,

Phosphatidase 10 .8～2%,

Glycerol 1.5～3%,

Water is an amount of, adds to 100%;

And the pH value of said emulsion composition is 3.5～7.0;

This method may further comprise the steps:

2. according to the method for claim 1, it is characterized in that in following (a) to (f) each or multinomial:

(a) pH value of said emulsion composition is 4.0～7.0;

(b) said emulsion composition comprises:

Vitamin K1 0.75-1.5%,

Soybean oil 8～12%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.75～2.5%,

Water is an amount of, adds to 100%;

(c) said emulsion composition comprises:

Vitamin K1 0.9-1.1%,

Soybean oil 9～11%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.95～2.45%,

Water is an amount of, adds to 100%;

(d) said emulsion composition comprises:

Vitamin K1 1.0%,

Soybean oil 10%,

Phosphatidase 11 .2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%;

(e) said emulsion composition is measured according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two the 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 70% is below 1.0 μ m;

(f) said emulsion composition particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is less than 20% greater than the breast grain of 1 μ m.

3. emulsion composition, it comprises:

Vitamin K1 0.5-2%,

Soybean oil 7.5～15%,

Phosphatidase 10 .8～2%,

Glycerol 1.5～3%,

Water is an amount of, adds to 100%.

4. according to the emulsion composition of claim 3, its pH value is 3.5～7.0.

5. according to the emulsion composition of claim 3, it comprises:

Vitamin K1 0.75-1.5%,

Soybean oil 8～12%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.75～2.5%,

Water is an amount of, adds to 100%.

6. according to the emulsion composition of claim 3, it comprises:

Vitamin K1 0.9-1.1%,

Soybean oil 9～11%,

Phosphatidase 11 .0～1.5%,

Glycerol 1.95～2.45%,

Water is an amount of, adds to 100%.

7. according to the emulsion composition of claim 3, it comprises:

Vitamin K1 1.0%,

Soybean oil 10%,

Phosphatidase 11 .2%,

Glycerol 2.2%,

Water is an amount of, adds to 100%.

8. according to the emulsion composition of claim 3, it is measured according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two the 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 70% is below 1.0 μ m.

9. according to the emulsion composition of claim 3, its particle diameter greater than with the breast grain sum that equals 0.5 μ m in, particle diameter is less than 20% greater than the breast grain of 1 μ m.

10. according to each emulsion composition of claim 3 to 9, it is characterized in that in following (a) to (e) each or multinomial:

(a) it comprises the vitamin K1 of 0.5-2%;

(b) it comprises 7.5～12.5% soybean oil;

(c) it comprises 0.8～2% phospholipid;

(d) it comprises 1.5～3% glycerol;

(e) it is the oil-in-water type injectable emulsion.