CN102552134B - Fat emulsion containing vitamin K1 - Google Patents

Abstract

The invention relates to a fat emulsion containing vitamin K1, and specifically relates to an emulsion composition comprising 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 invention also relates to a preparation method of the emulsion. The emulsion provided by the invention can be used medicinally.

Description

Lipomul containing vitamin K1

Technical field

The invention belongs to medical art, relate to a kind of lipomul, particularly relate to a kind of lipomul containing vitamin K1.The invention further relates to the preparation method preparing lipomul of the present invention.

Background technology

Vitamin K1 is the thick liquid of a kind of Yellow-to-orange clarification, odorless or almost odorless, meets light and easily decomposes.Vitamin K1 is easily molten in chloroform, ether or vegetable oil, slightly molten in ethanol, insoluble in water.The chemical name of vitamin K1 is: the mixture of the trans and cis-isomer of 2-methyl-3-(3,7,11,15-tetramethyl-2-hexadecene base)-Isosorbide-5-Nitrae-naphthalenedione, its molecular formula: C31H46O2, molecular weight: 450.71, and structural formula is:

Vitamin K1 is a kind of fatsoluble vitamin, is extensively present in green plants, exists with the form of phylloquinone (phylloquinone), also can synthetic.Vitamin K1 is the yellow oily liquid of purifying out from alfalfa the earliest, and called after vitamin K1, people just utilize vitamin K1 to do hemorrhage very early.The medical value of its hemostasis found in the thirties in 20th century.The conspicuous Garrick of biochemist of Denmark reaches the disease of nurse research chicken internal hemorrhage, he finds chicken haemophilia when injured, cannot condense, until add 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 gradually applied to various hemorrhage.Through basic research for many years and Clinical practice, the pharmacologically active beyond 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, therefore is widely used in clinical, and existing vitamin K1 is clinical commonly used drug.

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

Human body can not synthesize or secrete vitamin K1, and required vitamin K1 is mainly derived from food, and the escherichia coli in intestinal can synthetic vitamin K1 be also that human body utilizes.Vitamin K1 is the main component of prothrombing in composition blood of human body, can promote that liver manufactures thrombinogen, when organizing hemorrhage, platelet is destroyed, discharge thrombokinase and calcium ion, thrombinogen can with thrombokinase and calcium binding, change thrombin into, thrombin can catalysis plasma soluble conversion of fibrinogen be insoluble fibrin, reaches haemostatic effect.

Vitamin K1 can be removed smooth muscle spasmodic and shrink, scavenging activated oxygen, and has parahormone sample antiinflammatory action, hinders the release of the chemical mediators such as histamine, alleviate respiratory mucosa inflammation, minimizing is oozed out, blood vessel dilating, improve blood fortune, alleviate load before and after heart, correct heart failure; Excited respiratory center, suppresses cerebral cortex, improves brain microcirculation and prevent 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, thus suppresses growth of pathogenic bacteria, also has good curative effect to pertussis.

Vitamin K1 can directly act on multiple visceral smooth muscle, the internal organs pain stopped caused by multiple illness by the smooth muscle of lax spasm, as biliary colic, renal colic, 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 mostly clinically middlely is neonate because vitamin K1 lacks hemorrhage patient.Neonate birth 1 ~ 2 day diet is limited, the intake of vitamin K is few, in breast milk, vitamin content is low, in addition aseptic in intestinal, affects the synthesis of vitamin K, cause vitamin K deficiency, liver is not mature enough, and the process of liver synthesis prothrombin, VII, IX, X is obstructed, cruor time extending, cause hemorrhage, this disease is hemorrhagic disease of the newborn.The generation of hemorrhagic disease of the newborn (HDN) and avitaminous relation is just described as far back as nineteen fifty-two Dam etc.Three kinds can be divided into: Early onset hemorrhage, typical hemorrhage, delayed according to disease time.Be deficient in vitamin in body when neonate is just born K1, Pure breast feeding time longer neonate can fast-developing lack to serious vitamin K1: children's can occur typical after birth for first week or in 2 months, occur delayed hemorrhage, and both of these case is fallen ill suddenly, easily cause intracranial hemorrhage damage, disability rate is high, mortality rate is also high.The sickness rate of China is up to 0.5 ~ 2%, and the sickness rate of Area in Yantai Region is up to 9.03%, and China is the hotspot of Vitamin K Deficiency Bleeding disease.Medical research shows, neonate birth gives vitamin K1 can prevent vitamin K1 shortage property hemorrhage.The route of administration of vitamin K1 also affects drug effect.Oral vitamin K1 only can prevent typical hemorrhage, invalid to delayed hemorrhage, Longhan discovery vitamin K1 sustainable more than 2 months with 2mg intramuscular injection drug effect, oral relatively poor.2000, Hengxian County of China promotes conventional application vitamin K1 intramuscular injection in neonate birth 24h, and once, result of study showed: baby's vitamin K deficiency number of hospitalized obviously reduces, and vitamin K deficiency patient declines.In addition, because human liver can synthesize thrombinogen, hepatic disfunction person or surgery large operation person, all need to give a certain amount of vitamin K1 and can prevent hemorrhage.

Infantile respiratory disease occupies pediatric disease sickness rate 70%, and winter-spring season sickness rate is higher, and main manifestations is pneumonia, asthma, bronchitis, pertussis.

Infantile pneumonia refers to the pneumonia caused by various different pathogens and other factors, causes Pulmonary microcirculatory disturbance, and the reasons such as anoxia, infection, heating make blood present highly enriched, high gathering, high viscous pasty state, can cause and increase the weight of microcirculation disturbance.Vitamin K1 can change (cAMP/cGMP) ratio in cell, removes smooth muscle spasmodic and shrinks, scavenging activated oxygen, and there is parahormone sample antiinflammatory action, hinder the release of the chemical mediators such as histamine, alleviate respiratory mucosa inflammation, minimizing is oozed out, and reaches cough-relieving and to cough the effect of relievining asthma, vitamin K1 is expansible blood vessel also, improves blood fortune, alleviates load before and after heart, correct heart failure, excited respiratory center, suppress cerebral cortex, improve brain microcirculation and prevent cerebral edema, airway resistance can be reduced, increase alveolar ventilation, alleviate the ventilation ventilation exhaustion of crisis life, be conducive to exhaling the correction that declines, therefore vitamin K1 can be alleviated and improve the clinical symptoms of infantile pneumonia, also greatly can reduce the case fatality rate of severe pneumonia simultaneously, shorten the hospital stays, to aminophylline, promethazine, dexamethasone, the measure nonresponders such as oxygen uptake apply again, still promising result can be received, severe pneumonia of infants 100 example is assisted a ruler in governing a country by vitamin K1 treatment 50 example by someone, obtain good effect.

Vitamin K1 treatment bronchiolitis and bronchial asthma are reported also many, and domestic have research from 41 routine children asthmatic brouchitis case intravenous applications vitamin K1s treatment asthmatic bronchitiss of institute in October in October, 1998 to calendar year 2001, satisfactory effect.After child vein instillation vitamin K1, infant is quiet rapidly, falls asleep very soon, and cough with asthma obviously reduces, and breathes smooth-going.

Vitamin K1 also has good curative effect to pertussis.Pertussis is the Acute respiratory infectious disease caused by bordetella pertussis.Infectiousness is strong, is more common in infant, with liver source property blood coagulation disorders during pertussis outbreak, causes the petechial hemorrhage of respiratory mucosa, and this generation paroxysmal spasmodic cough of the cause of disease, 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, thus suppresses growth of pathogenic bacteria.

Within 1974, animal experiment study proves, vitamin K1 shrinks the intestinal that excised tea fresh leaves shrinks automatically and acetylcholine causes obvious inhibitory action, in recent years clinical trial shows: vitamin K1 can directly act on multiple visceral smooth muscle, by the smooth muscle of lax spasm, and the internal organs pain stopped caused by multiple illness, as biliary colic, renal colic, ureter and cystolith angor etc., effective percentage can reach 80 ~ 90%, and clinical vitamin K1 treatment children's gastrointestinal convulsion stomachache also has good effect.

Vitamin K1 can be used for treatment or prevents the disease relevant with osteoporosis or disease.Nineteen sixty, the people such as Bouckaert report that vitamin K can promote the union of fracture of rat and rabbit.Pettifor in 1975 etc. find that pregnant woman oral anticoagulant produces baby nose dysosteogenesis, the impact that reported first vitamin K deficiency is grown human bone, the hypothesis of vitamin K participant bone metabolism; Pettifor thinks that vitimin supplement K can promoting bone growing, reduce bone catabolism, positive preventive and therapeutic action is had to osteoporosis, its mechanism of action is to be converted into γ-Hydroxy GIutaminic Acid residue (Gla) as the glutaminic acid residue in some protein of cofactor catalysis of enzyme, this albuminoid is also called vitamin K dependent protein matter, and the γ-Hydroxy GIutaminic Acid residue after its activation has in conjunction with Ca ⁺⁺characteristic, thus make vitamin k-dependent protein play its biological activity, work as vitamin K deficiency, this proteinoid hydroxylation obstacle, therefore its biological activity also reduces.More bases and clinical trial describe vitamin K and osteoporotic relation.What separately have vitamin K in anticoagulant body capable of blocking recycles the clear proof causing vitamin K deficiency.Within 1975, have clinical report anticoagulant to the side effect of bone metabolism, after this crowd and animal experiment are constantly confirmed, after long diaphysis bone, and the too early calcification of end growth hone lamella.More than test and all point out vitamin K deficiency can disturb osseous tissue homergy.

Vitamin K1 contributes to treatment or the prevention of liver inflammatory disease.Research confirms that vitamin K1 can strengthen the effect of pathological changes Activity of hepatocytes and absorbability.Also have report to adopt megavitamin K " to swash and raise therapy " and treat hepatopathy, vitamin K can promote the Activity of hepatocytes of impending death, strengthens the absorbing power to nutrient substance, accelerates histiocytic recovery.

Version Chinese Pharmacopoeia in 2010 has recorded vitamin K 1 injection (see version Chinese Pharmacopoeia in 2010 two the 910th page), the aquesterilisa dispersion liquid in yellow liquid of this to be a kind of principal agent concentration be 10mg/ml, it allows micro-aobvious muddiness, need antifreeze preservation, if there is oil droplet to separate out or layering, 70-80 DEG C can be heated under dark conditions, jolting makes its natural cooling, if clarity normally still can continue to use, these information show that the injection that this pharmacopeia is recorded is a kind of injection using solubilizing agent that medicine is directly dissolved in the water, instead of the Emulsion that art of pharmacy is understood usually, because cannot by being heated to 70-80 DEG C and clinical acceptable clarity can being reached after jolting after the precipitation of Emulsion generation oil droplet or layering.

WO2011/153513A2 discloses the micro emulsion composition of fatsoluble vitamin K, allegedly vitamin K1 injectable emulsion (the PhytonadioneInjectable Emulsion of the alternative American Pharmacopeia of this micro emulsion composition, USP, can referred to as PIE-USP, its former name is vitamin K 1 injection) for clinical treatment.PIE-USP is a kind of aqueous dispersions, and wherein containing polyoxyethylated fatty acid (being also called Cremophor), this is a kind of potent surfactant.Although nomenclature of drug is recited as injection breast, the former product that grinds of vitamin K1 injectable emulsion (USP) is Merck & Co., Inc. it in fact not containing any oil, is therefore different from the vitamin K1 lipomul of indication of the present invention.Other manufacturer also uses the surfactant being similar to Cremophor to carry out solubilising vitamin K1, such as, use the HCO-60 of high hlb.The shortcoming using this type of surfactant is described in WO2011/153513A2, and by not using this type of reagent, but preparation one microemulsion is to substitute existing PIE-USP, this microemulsion such as containing 0.2 ~ 1% vitamin K1,0.5 ~ 2% soybean oil, 0.5 ~ 2% MCT, 4 ~ 25% phospholipid and 10% sucrose, phospholipid be the 13-25 of vitamin K1 amount doubly.But this microemulsion formed with the oil of relatively large phospholipid and small amount is a kind of metastable state being in critical state between solution and Emulsion, it easily departs from this metastable state because of outside cause such as pressure sterilizing, such as in the preparation embodiment of WO2011/153513A2, need with 0.22 μm of membrane filtration degerming.In addition, be favourable containing a certain amount of oil such as soybean oil, the such as soybean oil of 10% amount, its can for human body provide cannot or inconvenience picked-up energy.Be sufficient proof compared to the safety of oil for injection (such as safflower oil, the Fructus Canarii albi wet goods) soybean oil of other kind and effectiveness, employ many decades clinically with heavy dose of soybean oil that fat emulsion injection form provides.

People still expect to have new vitamin K 1 injection such as vitamin K1 fat emulsion injection for clinical.

Summary of the invention

The object of the invention is to provide a kind of new vitamin K 1 injection such as vitamin K1 fat emulsion injection for clinical, this injection has the beneficial effect of at least one such as described in the present invention.The present inventor finds, the vitamin K 1 injection prepared by the special formulation comprising about 10% soybean oil is the fat milk that a kind of performance can meet clinical application demand completely.The present invention is based on this find and be accomplished.

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

Vitamin K1 0.5-2%,

Soybean oil 7.5 ~ 15%,

Phosphatidase 0 .8 ~ 2%,

Glycerol 1.5 ~ 3%,

Water is appropriate, adds to 100%.

Emulsion composition according to a first aspect of the present invention, its pH value is 7.0 ~ 9.0.In one embodiment, pH value is 7.0 ~ 8.5.In one embodiment, pH value is 7.5 ~ 8.5.

Emulsion composition according to a first aspect of the present invention, its method of pressing under Chinese Pharmacopoeia version in 2010 two annex VI H items measures, and pH value is 7.0 ~ 9.0.In one embodiment, pH value is 7.0 ~ 8.5.In one embodiment, pH value is 7.5 ~ 8.5.

Emulsion composition according to a first aspect of the present invention, wherein also comprises appropriate pH adjusting agent, and regulates the pH value of this emulsion composition to be 7.0 ~ 9.0.In one embodiment, pH value is 7.0 ~ 8.5.In one embodiment, pH value is 7.5 ~ 8.5.In one embodiment, described pH adjusting agent is sodium hydroxide and/or hydrochloric acid.

Emulsion composition according to a first aspect of the present invention, it comprises:

Vitamin K1 0.75-1.5%,

Soybean oil 8 ~ 12%,

Phosphatidase 1 .0 ~ 1.5%,

Glycerol 1.75 ~ 2.5%,

Water is appropriate, adds to 100%.

Emulsion composition according to a first aspect of the present invention, it comprises:

Vitamin K1 0.9-1.1%,

Soybean oil 9 ~ 11%,

Phosphatidase 1 .0 ~ 1.5%,

Glycerol 1.95 ~ 2.45%,

Water is appropriate, adds to 100%.

Emulsion composition according to a first aspect of the present invention, it comprises:

Vitamin K1 1.0%,

Soybean oil 10%,

Phosphatidase 1 .2%,

Glycerol 2.2%,

Water is appropriate, adds to 100%.

Emulsion composition according to a first aspect of the present invention, it measures according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two annex IY E granularities and particle size distribution method the 3rd method (light scattering method), and the particle diameter of the newborn grain of more than 70% is below 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 80% is below 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 90% is below 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 70% is between 0.01 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 80% is between 0.01 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 90% is between 0.01 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 70% is between 0.05 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 80% is between 0.05 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 90% is between 0.05 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 70% is between 0.1 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 80% is between 0.1 ~ 1.0 μm.In one embodiment, the particle diameter of newborn grain of more than 90% is between 0.1 ~ 1.0 μm.In one embodiment, be greater than and equal in the newborn grain sum of 0.5 μm at particle diameter, the newborn grain that particle diameter is greater than 1 μm is less than 20%, is preferably less than 15%, is less than 10%, is less than 5%.

Emulsion composition according to a first aspect of the present invention, it is according to high performance liquid chromatography described in Chinese Pharmacopoeia 2010 editions two annex V D, measure with following liquid phase chromatogram condition, the amount containing vitamin K1 in every 1ml is 5 ~ 20mg (such as 7.5 ~ 15mg, such as 9 ~ 11mg):

(1) chromatographic condition and system suitability test: be filler with octadecylsilane chemically bonded silica, with the ethanol-water mixture of volume ratio 95: 5 for mobile phase, column temperature is 35 DEG C, and determined wavelength is 254nm, and theoretical cam curve calculates should be not less than 1500 with vitamin K1;

(2) preparation of need testing solution: it is appropriate that precision measures described compositions, adds isopropyl alcohol and dissolves and quantitatively make the solution being about 0.05mg in every 1ml containing vitamin K1, as need testing solution;

(3) preparation of reference substance solution: take vitamin K1 reference substance appropriate, accurately weighed, (in one embodiment, this 10% fat emulsion injection is method according to Examples below 1 of the present invention but not 10% fat emulsion injection prepared of vitaminize K1 to add 10% fat emulsion injection; In one embodiment, this 10% fat emulsion injection is 10% commercially available fat emulsion injection) 1ml, add isopropyl alcohol and dissolve and quantitatively make the solution being about 0.05mg in every 1ml containing vitamin K1, product solution in contrast;

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

Emulsion composition according to a first aspect of the present invention, it measures by following method, and in every 1ml, glycerinated amount is 15 ~ 30mg (such as 17.5 ~ 25mg, such as 19.5 ~ 24.5mg):

Precision measures described compositions 2.5ml, puts in conical flask, and add water 100ml, adds bromocresol purple indicator solution 5, shakes up, if aobvious acid, drip 0.1mol/L sodium hydroxide solution, makes solution be bluish violet; If aobvious alkalescence, first should drip 0.5mol/L sulfuric acid solution and be adjusted to solution just in yellow, dripping 0.1mol/L sodium hydroxide solution again makes solution be bluish violet, add the 0.7% Potassium metaperiodate. solution 100ml faced with newly joining, put in the water-bath of 37 ~ 40 DEG C and be incubated 15 minutes, and jolting constantly, add 1,2-propylene glycol 3ml, place 5 minutes, be titrated to solution just in bluish violet with 0.1mol/L sodium hydroxide titration liquid, obtain the amount of sodium hydroxide titration liquid used, the C3H8O3 being equivalent to 9.210mg with the 0.1mol/L sodium hydroxide titration liquid of every 1ml calculates glycerol content.

Emulsion composition according to a first aspect of the present invention, it is according to high performance liquid chromatography described in Chinese Pharmacopoeia 2010 editions two annex V D, measure with following liquid phase chromatogram condition, the amount containing triglyceride in every 1ml is 75 ~ 125mg (such as 80 ~ 120mg, such as 90 ~ 110mg):

(1) chromatographic condition and system suitability: be filler with silica gel, be that the normal hexane-isopropyl alcohol-acetic acid mixture of 98.9: 1: 0.1 is for mobile phase with volume ratio, evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 DEG C) detect, 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 solution: get triglyceride and each 10mg of oleic acid, in 25ml measuring bottle, dissolve with mobile phase and be diluted to scale, shaking up, to obtain final product;

(3) preparation of reference substance solution: get soybean oil reference substance and be about 0.35g, accurately weighed, put in 50ml measuring bottle, hold mixed solution with normal hexane and isopropyl alcohol etc. dissolve and be diluted to scale, be stock solution, precision measures stock solution 3ml and 4ml, put in 25ml measuring bottle respectively, 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 measures described compositions 4ml, puts in 50ml measuring bottle, holds mixed solution and dissolves and be diluted to scale, shake up with normal hexane and isopropyl alcohol etc.; Precision measures 3ml, puts in 25ml measuring bottle, is diluted to scale with mobile phase, shake up, to obtain final product;

(5) algoscopy: precision measures reference substance solution 1, reference substance solution 2 respectively, each 10 μ l of need testing solution, in alter least-squares chromatograph of liquid, obtain calculated by peak area through chromatographic isolation.

Emulsion composition according to a first aspect of the 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 the vitamin K1 of about 1.0%.

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

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

Emulsion composition according to a first aspect of the present invention, it comprises the glycerol of 1.5 ~ 3%, preferably comprises the glycerol of 1.75 ~ 2.5%, preferably comprises the glycerol of 1.95 ~ 2.45%, preferably comprises the glycerol of about 2.2%.

Emulsion composition according to a first aspect of the present invention, it is oil-in-water type injectable emulsion.

Second aspect present invention provides the method preparing emulsion composition described in first aspect present invention, and it comprises the following steps:

(1) vitamin K1, soybean oil, phospholipid, glycerol is taken by recipe quantity, for subsequent use;

(2) water for injection preparation total amount being about 60-90% (such as about 80%) adds stainless cylinder of steel, adds the glycerol of recipe quantity, stirs, obtained aqueous phase;

(3) soybean oil of recipe quantity is added in another stainless cylinder of steel, pass into nitrogen current, add vitamin K1 and the phospholipid of recipe quantity, be stirred to each material dissolution, obtained oil phase;

(4) pass into nitrogen current, oil phase is slowly added in the aqueous phase of high-speed stirred, continue high-speed stirred in good time, obtain colostrum;

(5) regulate the pH value of this colostric fluid to prescribed limit with sodium hydroxide solution, then inject and be settled to theoretical recipe quantity with water, stir, regulate solution ph to prescribed limit with sodium hydroxide or hydrochloric acid solution if desired;

(6) under nitrogen flowing, make colostrum high pressure homogenization under 40-70MPa (such as 55-60MPa) pressure, then under 5-25MPa (such as 12-18MPa) pressure low pressure homogenize, cooling, then use the membrane filtration of 10 μm;

(7) filtrate fill is in ampoule bottle, fills nitrogen sealing by fusing, pressure sterilizing, to obtain final product.

Method according to a second aspect of the present invention, it is included in suitable processing step, adds other adjuvants.In one embodiment, in step (3), stabilizing agent is added with vitamin K1 and phospholipid.In one embodiment, isotonic agent is added after obtaining colostrum in step (4).

Method according to a second aspect of the present invention, if also comprise other adjuvant in emulsion composition, such as coemulsifier, stabilizing agent etc., these adjuvants can prepared aqueous phase according to its hydrophilic or lipophile or add when preparing oil phase.

Method according to a second aspect of the present invention, in compositions encapsulation and measure before sterilizing and or regulate the pH value of said composition to be very easy; And (namely becoming) should not regulate the pH value of said composition usually again after compositions encapsulation also sterilizing.Those skilled in the art understand, before sterilization, the pH value of compositions may change to some extent, or increase a little or reduce a little, when therefore pH value being regulated in dosing process, suitably can depart from target ph, the pH value after compositions sterilizing be fallen into as far as possible or near object pH value or scope.In this sense, in the present invention, when mentioning the pH value of emulsion composition, if do not illustrated in addition, the pH value after referring to sterilization treatment; Such as in hereafter preparation example 1, when describing composite formula, mention that " pH adjusting agent is appropriate; adjust pH to 8.0 " refers in formula or to add appropriate pH adjusting agent in dosing process appropriate, make medicinal liquid through corresponding preparation method process and obtain after final emulsion composition through pressure sterilizing, the pH value of this emulsion composition is about 8.0, and such as pH value is in the scope of 8.0 ± 0.2, and such as pH value is in the scope of 8.0 ± 0.1.

The documents and materials of the present invention's citation, its full content is incorporated to herein by reference.In either side of the present invention, any one or more technical characteristics of its arbitrary embodiment can be combined in other embodiment arbitrary of this aspect, also can be combined in arbitrary embodiment on the other hand, as long as this combination there will not be contradiction.

The various terms that the present invention uses have the general sense that those skilled in the art's routine is understood, and when inconsistent with this general sense, are as the criterion with the present invention.

In the present invention, for writing conveniently, vitamin K1 simply can be written as VK1.Those skilled in the art know, and Menaquinone K6 (can be abbreviated as VK2) is a kind of material similar to VK1 in structure, physicochemical property and biologic activity etc.

The present invention provides a kind of fluid composition generally, and main carriers is wherein water, and solute comprises liquid substance (such as glycerol) and/or solid matter (such as in semi-solid Ovum Gallus domesticus Flavus lecithin).Therefore, for simplicity, in the present invention, when using " % " when indicated concentration percent, if not otherwise indicated, weight/volume percent is referred to; Such as.For the formula containing " vitamin K1 be 1%, soybean oil 10%, Ovum Gallus domesticus Flavus lecithin 1.2%, glycerol 2.2%, water add to 100% in right amount ", its refer in every 100ml formula containing vitamin K1 be 1g, soybean oil 10g, Ovum Gallus domesticus Flavus lecithin 1.2g, glycerol 2.2g and add to the water of 100ml volume.

It will be appreciated by those skilled in the art that newborn grain particle diameter normally statistical way calculate or provide.Such as, phrase " particle diameter of the newborn grain of more than 70% is below 1.0 μm " can refer to and measure the particle diameter of the newborn grain particle of some according to (such as microscopic method, light scattering method etc.) someway, such as measure about 1000 newborn grain particles, then particle diameter is dropped on these about 1000 populations divided by statistical computation of the population in less than 1.0 μm scopes, the percent obtained is greater than 70%.Other similar terms also has similar meaning.

In the present invention, phrase " newborn grain particle diameter is at about 0.5 μm ", if not otherwise indicated, refers to that the particle diameter of newborn grain is between 0.2 μm ~ 1.0 μm.

In the present invention, term " 10% fat emulsion injection ", if not otherwise indicated, refer to according to method of preparing emulsion, substantially can intravenous Emulsion by what comprise that following formula prepares:

Soybean oil 10%,

Ovum Gallus domesticus Flavus lecithin 1.2%,

Glycerol 2.2%,

Water is appropriate, adds to 100%.

The present invention is the emulsion composition of injection, and wherein each kind of material is 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 usually used as oil phase solvent, and it also has the effect providing energy certainly.Usually play the effect of emulsifying agent at emulsion composition mesolecithal lecithin of the present invention, and glycerol normally plays the effect of osmotic pressure regulator; Further, water for injection uses as aqueous phase solvent.

Certainly, the adjuvant that the present invention is used, comprises soybean oil, and being needs to meet necessarily required, and such as, when preparing fat emulsion for injection, these adjuvants should can be used for injection to use.This well known to a person skilled in the art.

Phospholipid of the present invention is selected from following refining phospholipid: Ovum Gallus domesticus Flavus lecithin, soybean lecithin, cephalin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline etc.They commercially, also can use usual method, as Batteries by Organic Solvent Partition Process is prepared.Namely raw phospholipid is dissolved in cold n-hexane-acetone, under stirring, slowly adds acetone, filtered and recycled insoluble matter, after repeating this operation, steam solvent and just can obtain refined phosphatide.Its Main Ingredients and Appearance comprises phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE etc.Phospholipid as other also can be illustrated out phosphatidylinositols, Phosphatidylserine, sphingomyelins etc.The phospholipid preferably Ovum Gallus domesticus Flavus lecithin of indication of the present invention.

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

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

As the medium chain fatty glyceride that coemulsifier can also be containing 50g or following amount in every 1000ml lipomul, said medium chain fatty glyceride has C ₈~ C ₁₂the ester that the fatty acid of chain length and glycerol are formed, diester or three esters, or the mixture of these esters, the preferably triglyceride of the sad and capric acid of satisfied fatty acid, or its commercial product, as Miglyol812 etc.

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

The optional albumin of osmotic pressure regulator, dextrin, ethene polymers, non-ionic surface active agent, gelatin, hetastarch, propylene glycol, glycerol etc., make the osmotic pressure ratio of the lipomul made (assay method carries out according to " Chinese Pharmacopoeia version in 2010 two annex IX G " osmotic pressure molar density algoscopy " ") be 0.5 ~ 3, 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 version in 2010 two.

The pH adjusting agent that the present invention uses includes but not limited to sodium hydroxide solution, hydrochloric acid solution, such as 1M sodium hydroxide solution, 1M hydrochloric acid solution.In the present invention, when mentioning emulsion composition of the present invention or the pH value for the compositions that contrasts, this pH value is the final ph of compositions, instead of the pH value of its intermediate material.Such as Emulsion of the present invention, the pH value measured after the pH value measured before being dispensed in ampoule bottle and sterilizing likely has slight variations, the pH value mentioned in the present invention refer to sterilizing after the pH value of Emulsion.Therefore in production practices, when preparing burden before sterilization, can pH value is possible before and after with due regard to this sterilizing change; If such as pH value can reduce by 0.2 unit after sterilization, and target ph is 8.0, then the pH value of the emulsion composition prepared before sterilizing is adjusted to 8.2 and is advisable.

Lipomul provided by the invention can be prepared by the following method:

Vitamin K1, soybean oil, phospholipid, glycerol is taken by recipe quantity, for subsequent use; The water for injection of preparation total amount about 80% is added stainless cylinder of steel, adds the glycerol of recipe quantity while stirring; The soybean oil of recipe quantity is added in another stainless cylinder of steel, pass into nitrogen current, add vitamin K1 and the phospholipid of recipe quantity, be stirred to phospholipid and dissolve; Pass into nitrogen current, oil phase is slowly added in the aqueous phase of high-speed stirred, continue high-speed stirred in good time, regulate solution ph to prescribed limit with sodium hydroxide solution, then this colostrum is settled to theoretical recipe quantity, stir; Under nitrogen flowing, make colostrum high pressure homogenization under 40-70MPa (such as 55-60MPa) pressure, then under 5-25MPa (such as 12-18MPa) pressure low pressure homogenize, cooling, then use the membrane filtration of 10 μm; Filtrate fill, in ampoule bottle, is filled nitrogen sealing by fusing, pressure sterilizing, to be obtained final product.

According to the needs in preparation, in suitable processing step, such as, after colostrum is formed, other adjuvants can be added, as stabilizing agent, isotonic agent etc.

Lipomul containing vitamin K1 of the present invention is suitable for medicinal, and such as oral and injection uses.Especially, emulsion composition of the present invention can use in suitable injection administration, and newborn grain diameter is wherein little, and mean diameter between 0.2 ~ 1 μm, and has and meets the general prescription of medicine.

Lipomul containing vitamin K1 of the present invention by the approach of injection and mode medication, can comprise approach and the modes such as intramuscular injection, intravenous injection, intravenous drip.Such as, when giving people the used time, the lipomul containing vitamin K1 of the present invention, dosage for each person every day can be 0.1mg ~ 50mg, preferably 1mg ~ 40mg.Can once-a-day or multiple dosing, also can be administered once for 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 as normal saline, glucose solution, xylitol solution, fructose soln, woods grignard T ₃with glycerol etc.Especially, described transfusion diluent is commercially available 10% fat emulsion injection for nutritional supplement.

The lipomul that the present invention contains vitamin K1 has good pharmaceutics performance, meets medicinal requirements.Such as, the vitamin K1 emulsion composition that the embodiment of the present invention 1 prepares is tested about the specification of the anaphylaxis of injection systemic administration Emulsion, hemolytic and vascular stimulation test, muscle irritation, subcutaneous irritation test according to state food and drug administration, and result shows: the vitamin K1 fat emulsion injection of 4mg/kg and 1mg/kg to Cavia porcellus without allergenic effect; Vitamin K1 fat emulsion injection 0.2mg/kg ~ 1mg/kg to family's rabbit erythrocyte without obvious hemolysis in vitro and agglutination; 1mg/ml vitamin K1 fat emulsion injection 1mg/min intravenously administrable, every day 1 time, continuous 7 days, to rabbit auricular vein without obvious stimulation effect; The vitamin K1 fat emulsion injection subcutaneous injection 1ml of 10mg/ml, every day 1 time, continuous 7 days, without obvious stimulation effect.

Detailed description of the invention

Following examples further illustrate the present invention, instead of restriction the present invention.In example below, when using pH adjusting agent and regulating material, unless otherwise noted, regulate with 1M sodium hydroxide solution or 1M hydrochloric acid solution, its consumption be make the pH value of material particularly solution be adjusted to setting (such as indicated value ± 0.1 scope in) or scope, such as, when the pH value of current material solution is 6.5, target ph is 8.0, first be adjusted to 8.0 ± 0.1 with 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.The hereafter object of preparation process in order to illustrate, and based on each citing comparability and make some specific description, those skilled in the art can therefrom summarize the method obtaining pharmaceutical compositions of the present invention completely according to existing knowledge.

a, compositions preparation 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,

PH adjusting agent is appropriate, adjust pH to 8.0,

Water is appropriate, adds to 100ml.

Method for making:

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

(2) preparation of aqueous phase: the water for injection of preparation total amount about 80% is added stainless cylinder of steel, adds the glycerol (injection) of recipe quantity while stirring.

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

(4) preparation of colostrum: pass into nitrogen current, oil phase is slowly added in the aqueous phase of high-speed stirred, continue high-speed stirred after at least 5 minutes, solution ph is regulated (on setting basis, suitably to increase 0.1 pH unit at setting with 1M sodium hydroxide solution, the pH value after Emulsion sterilizing is made to reach the value of regulation), again colostrum is settled to theoretical amount, stirs.

(5) nitrogen current is passed into, colostrum is under 55-60MPa pressure after high pressure homogenize 4 times, 12-18MPa low pressure homogenizing 1 time (making the newborn grain granularity of the overwhelming majority (microscopic examination has more than 95%) between 0.2 ~ 0.8um) cools afterwards, then uses the membrane filtration of 10 μm.

(6) filtrate fill is in 1ml brown glass ampoules bottle, sealing by fusing after inflated with nitrogen, and 118 DEG C of moist heat sterilization 25min, to obtain final product.

As a kind of medicine, after this can further by this sterilising prods labeling after lamp inspection, packaging, after the full review of sampling is qualified and get final product.In addition as medicine, the capacity of ampoule bottle can be changed, to provide the single-dose preparations of various dose.

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,

PH adjusting agent is appropriate, adjust pH to 7.5,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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,

Glycerol 2.2g,

PH adjusting agent is appropriate, adjust pH to 8.5,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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,

PH adjusting agent is appropriate, adjust pH to 8.0,

Water is appropriate, adds to 100ml.

Method for making: the vitamin K1 in prescription, soybean oil, Ovum Gallus domesticus Flavus lecithin and glycerol are heated at 70 ~ 80 DEG C and abundant mix homogeneously, 20ml water for injection is added in the solution obtained, stir, inject and make it into 100ml with water, with this solution of homogenizer preliminary homogenisation, adjust ph is to setting if desired.Then, under the pressure of 1100Bar, by NS1001L type high pressure homogenizer (Italian GEA Niro Soavi company) homogenize 7 times (making most newborn grain granularity between 0.2 ~ 0.8um), containing vitamin K1 in obtained every 1ml is the lipomul of 10mg.This lipomul can be potted in little ampoule bottle, often props up and can fill 1ml, 2ml, 5ml, 10ml, 20ml etc., then through 115 DEG C of sterilizings 30 minutes, obtains the lipomul injection containing vitamin K1.This injection can directly intramuscular injection or intravenous injection, also can instil with 5% glucose injection dilution posterior vein, can also with 10% fat emulsion injection dilution posterior vein instillation.

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 1.5g,

PH adjusting agent is appropriate, adjust pH to 8.0,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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 3g,

PH adjusting agent is appropriate, adjust pH to 8.0,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

preparation example 7: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 1.5g,

Soybean oil 8g,

Soybean lecithin 1.0g,

Glycerol 2.5g,

PH adjusting agent is appropriate, adjust pH to 8.0,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

preparation example 8: prepare emulsion composition of the present invention

Composite formula:

Vitamin K1 0.75g,

Soybean oil 12g,

Soybean lecithin 1.5g,

Glycerol 1.75g,

PH adjusting agent is appropriate, adjust pH to 7.0,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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 1.95g,

Propylene glycol 0.2g,

PH adjusting agent is appropriate, adjust pH to 7.5,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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 2.2g,

Oleic acid 0.25g,

PH adjusting agent is appropriate, adjust pH to 7.9,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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,

Miglyol 812 2.0g

PH adjusting agent is appropriate, adjust pH to 8.2,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

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,

Cholesterol 0.2g,

PH adjusting agent is appropriate, adjust pH to 8.5,

Water is appropriate, adds to 100ml.

Method for making: carry out with reference to the method recorded in preparation example 1.

Prepare the emulsion composition of 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 except 6.5, formula and method for making are with preparation example 1;

Reference examples 2: except pH value is except 5.8, formula and method for making are with preparation example 2;

Reference examples 3: except pH value is except 9.0, formula and method for making are with preparation example 3;

Reference examples 4: except pH value is except 9.5, formula and method for making are with preparation example 4;

Reference examples 5: except pH value is except 6.5, formula and method for making are with preparation example 5;

Reference examples 6: except pH value is except 5.7, formula and method for making are with preparation example 6;

Reference examples 7: except pH value is except 5.0, formula and method for making are with preparation example 7;

Reference examples 8: except pH value is except 9.0, formula and method for making are with preparation example 8;

Reference examples 9: except pH value is except 9.6, formula and method for making are with preparation example 9;

Reference examples 10: except pH value is except 10.2, formula and method for making are with preparation example 10;

Reference examples 11: except pH value is except 6.7, formula and method for making are with preparation example 11;

Reference examples 12: except pH value is except 9.1, formula and method for making are with preparation example 12.

Reference examples 13: except replacing with except VK2 by VK1, formula and method for making are with preparation example 1.

Reference examples 14: except replacing with except VK2 by VK1, formula and method for making are with preparation example 3.

Reference examples 15: except replacing with except VK2 by VK1, formula and method for making are with preparation example 7.

Reference examples 16: except replacing with except olive oil by soybean oil, formula 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 the hot water of 65 DEG C, heat and make it be mixed into oil phase uniformly.Separately 1.0g poloxamer F68, glycerol 2.2g are about 80ml dissolving with water for injection and make into aqueous phase.Then according to preparation example 1 step (4), (5) and (6) process, whole emulsion is obtained.

Reference examples 18: get 1g vitamin K1, soybean oil 2g and lecithin 1g, in nitrogen current, in the hot water of 65 DEG C, heats and makes it be mixed into oil phase uniformly.In addition 5g glucose is dissolved in the distilled water for injection of 80ml and becomes aqueous phase.Aqueous phase is added in oil phase while stirring, to be then placed in ice the ultrasound mulser emulsifying of limit cooling limit 20 minutes.The sodium hydroxide solution of the pH value 0.1N of gained emulsion is adjusted to 6.5, adds water for injection afterwards and make into 100ml.Then according to preparation example 1 step (5) and (6) process, whole emulsion is obtained.

b, detection method part: the method detecting 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 internal performance such as 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 detecting emulsion composition of the present invention

Method: according to the method described in Chinese Pharmacopoeia version in 2010 two annex VI H, the emulsion composition of the present invention that the present invention of direct mensuration obtains or other various sample.In the present invention's test, when providing pH value, be the average of 3 parallel assays.The pH value of some emulsion compositions prepared by the present invention is respectively see the value (and the difference between formula sign value is less than 0.1 pH unit) described in preparation example 1 ~ 12 formula.

detection side's rule 2: the granularity and the distribution that detect 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 annex IX E granularities and particle size distribution method the 3rd method (light scattering method), adopt laser diffraction particle size distribution instrument to carry out, record and add up the particle diameter being no less than 500 newborn grain particles.Calculate the percent (can represent with " percentile 1 " in this article) that the newborn grain number of particle diameter below 1.0 μm accounts for the total statistical number of newborn grain.Calculate simultaneously and to be greater than at particle diameter and to equal in the newborn grain sum of 0.5 μm, particle diameter is greater than the percent (can represent with " percentile 2 " in this article) of the newborn grain of 1 μm.

Generally speaking, as Emulsion that can be used for intravenous injection, the newborn grain number of particle diameter below 1.0 μm accounts for the percent of the total statistical number of newborn grain, and namely percentile 1 should more than 70%, preferably more than 80%, preferably more than 90%.In addition, generally speaking, as Emulsion that can be used for intravenous injection, be greater than at particle diameter and equal in the newborn grain sum of 0.5 μm, particle diameter is greater than the percent of the newborn grain of 1 μm, and namely percentile 2 should be less than 20%, is preferably less than 15%, is less than 10%, is less than 5%.

detection side's rule 3: the content detecting vitamin K1 in emulsion composition of the present invention

Method:

(1) measure according to high performance liquid chromatography (Chinese Pharmacopoeia 2010 editions two annex V D);

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

(3) preparation of need testing solution: it is appropriate that precision measures emulsion composition, adding that isopropyl alcohol dissolves and quantitatively to make in every 1ml containing vitamin K1 is the solution of 0.05mg, as need testing solution;

(4) preparation of reference substance solution: take vitamin K1 reference substance appropriate, accurately weighed, add 10% fat emulsion injection (to prepare with reference to preparation example 1 of the present invention, difference is non-vitaminize K1) 1ml, add isopropyl alcohol dissolve and quantitatively make in every 1ml containing vitamin K1 be the solution of 0.05mg, product solution in contrast;

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

The content of vitamin K1 can represent with % (w/v) or mg/ml, or represents with the percent being equivalent to formula labelled amount.

detection side's rule 4: the content detecting glycerol in emulsion composition of the present invention

Method: precision measures emulsion composition 2.5ml, puts in conical flask, and add water 100ml, adds bromocresol purple indicator solution 5, shakes up, if aobvious acid, drip 0.1mol/L sodium hydroxide solution, makes solution be bluish violet; If aobvious alkalescence, first should drip 0.5mol/L sulfuric acid solution and be adjusted to solution just in yellow, dripping 0.1mol/L sodium hydroxide solution again makes solution be bluish violet, adds 0.7% Potassium metaperiodate. solution (facing with newly joining) 100ml, puts in the water-bath of 37 ~ 40 DEG C and is incubated 15 minutes, and jolting constantly, add 1,2-PD 3ml, place 5 minutes, be titrated to solution just in bluish violet with 0.1mol/L sodium hydroxide titration liquid, obtain final product.The 0.1mol/L sodium hydroxide titration liquid of every 1ml is equivalent to the C3H8O3 of 9.210mg.

The content of glycerol can represent with % (w/v) or mg/ml, or represents with the percent being equivalent to formula labelled amount.

detection side's rule 5: the content detecting triglyceride in emulsion composition of the present invention

Method:

(1) measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 two annex VD);

(2) chromatographic condition and system suitability: 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 DEG C) detect, 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 solution: get triglyceride and each 10mg of oleic acid, in 25ml measuring bottle, dissolve with mobile phase and be diluted to scale, shaking up, to obtain final product;

(4) preparation of reference substance solution: get soybean oil reference substance and be about 0.35g, accurately weighed, put in 50ml measuring bottle, hold mixed solution with normal hexane and isopropyl alcohol etc. and dissolve and be diluted to scale, be stock solution; Precision measures stock solution 3ml and 4ml, puts respectively in 25ml measuring bottle, is diluted to scale with mobile phase, shake up, and is reference substance solution 1 and reference substance solution 2;

(5) preparation of need testing solution: precision measures emulsion composition 4ml, puts in 50ml measuring bottle, holds mixed solution and dissolves and be diluted to scale, shake up with normal hexane and isopropyl alcohol etc.; Precision measures 3ml, puts in 25ml measuring bottle, is diluted to scale with mobile phase, shake up, to obtain final product;

(6) algoscopy: precision measures reference substance solution 1, reference substance solution 2 respectively, each 10 μ l of need testing solution, in alter least-squares chromatograph of liquid, obtain calculated by peak area through chromatographic isolation, to obtain final product.

The content of triglyceride can represent with % (w/v) or mg/ml, or represents with the percent being equivalent to formula labelled amount.

detection side's rule 6: the content detecting emulsion composition free fatty acid of the present invention

The preparation of contrast solution: get Palmic acid 0.64g, accurately weighed, put in 500ml measuring bottle, add normal heptane and dissolve and be diluted to scale, shake up.

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

Precision measures diluent and each 1ml of contrast solution, puts respectively in 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), jolting 1 minute, places 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, leave standstill at least 15 minutes, make layering, precision measures upper liquid 3ml respectively, put in 10ml centrifuge tube, (get Nile blue 0.04g, add water 200ml, after making dissolving to add Nile blue indicator solution, add normal heptane 100ml jolting, discard upper strata normal heptane.Repeatable operation 4 times.Get lower aqueous solution 20ml, add dehydrated alcohol 180ml, mixing.This liquid is put in brown bottle, can deposit one month under room temperature.Under logical nitrogen condition, be titrated to solution with sodium hydroxide titration liquid (0.01mol/L) and show lavender.The milliliter number of need testing solution consumption sodium hydroxide titration liquid and reference substance solution consume the ratio of sodium hydroxide titration liquid milliliter number, then are multiplied by 5.0mmol/L, amount to into the concentration symbols of free acid.

c, test example part

test example 1: in investigation vitamin K1 formula, Semen sojae atricolor oil mass is on the impact of formula

With the emulsion composition of 6 different formulations preparations containing vitamin K1, its formula and method for making are see preparation example 1, difference is the amount of the soybean oil used, the amount of the soybean oil used in 6 formula is respectively the different samples of 0%, 2.5%, 5%, 7.5%, 10%, 15%, obtains colostrum and whole breast in step (4) described in preparation example 1 and step (6) respectively.The colostrum of different prescription is put in color comparison tube to leave standstill and observe delamination after 6 hours.Whole breast is put to observe delamination after 10000rpm × 10min centrifugal treating in centrifuge tube, and the method simultaneously according to " detection side's rule 2 " measures percentile 1 (" percent 1 ") newborn eventually and percentile 2 (" percent 2 ").The results are shown in following table:

Visible, the amount of the about 7-15% soybean oil using the present invention to advocate is useful for forming more stable emulsion.

test example 2: measure emulsion composition and at high temperature place the content of rear vitamin K1 and dissociate the content of fatty acid

12 emulsion compositions prepared by prepared by preparation example 1-12 12 emulsion composition of the present invention and reference examples 1-12 are divided in brown ampoule, be placed in 50 DEG C of baking ovens to place 2 months, then the content (mg/ml) of vitamin K1 is measured by the method for detection side's rule 3 mentioned above, measure the content without the vitamin K1 in the respective sample of high-temperature treatment in addition, calculate the remaining percent (%) that high temperature places vitamin K1 in reagent after 2 months thus.In addition, measure the content without high-temperature process and the free fatty through high-temperature process sample by the method for detection side's rule 6 mentioned above, calculate high temperature thus and to place after 2 months reagent free fatty acid corresponding to without the percent (%) during high-temperature process.The results are shown in following table.

Table

test example 3: 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 content tested the percent basis that formula indicates content.The results are shown in following table:

Reagent	VK1 content (%)	Reagent	VK1 content (%)
				Preparation example 1	99.7	Preparation example 7	99.1
Preparation example 2	98.9	Preparation example 8	99.8
				Preparation example 3	100.1	Preparation example 9	99.6
Preparation example 4	99.2	Preparation example 10	101.8
				Preparation example 5	99.7	Preparation example 11	100.2
Preparation example 6	100.3	Preparation example 12	99.9

2, glycerol content:measure according to method shown in above.The percent basis that formula indicates content is accounted for the content of test.The results are shown in following table:

Reagent	Glycerol content (%)	Reagent	Glycerol content (%)
				Preparation example 1	98.9	Preparation example 7	99.9
Preparation example 2	102.3	Preparation example 8	97.8
				Preparation example 3	100.7	Preparation example 9	101.3
Preparation example 4	99.8	Preparation example 10	100.2
				Preparation example 5	99.3	Preparation example 11	102.2
Preparation example 6	101.2	Preparation example 12	100.9

3, triglyceride content:measure according to method shown in above.Account for soybean oil in formula with the content of test and indicate the percent basis of content.The results are shown in following table:

Reagent	Triglyceride content (%)	Reagent	Triglyceride content (%)
				Preparation example 1	97.7	Preparation example 7	101.9
Preparation example 2	98.6	Preparation example 8	99.8
				Preparation example 3	104.7	Preparation example 9	97.3
Preparation example 4	97.8	Preparation example 10	104.5
				Preparation example 5	99.8	Preparation example 11	98.6
Preparation example 6	104.5	Preparation example 12	103.5

In addition, with above-mentioned three parameters and pH value, newborn grain granularity for index, 12 emulsion compositions of the present invention shading under the room temperature of about 25 DEG C investigating preparation example 1 to preparation example 12 preserves 24 months.Relatively 24 months time value and 0 month initial value between situation of change, result shows:

The pH value of (1) 12 sample is respectively compared with its initial value, and change is all less than 0.2 pH value unit, and all in the acceptable scope of the human body of pH value 7 ~ 9.

The newborn grain granularity of (2) 12 samples:

When 0 month, the newborn grain particle diameter of more than 85% is at about 0.5 μm; Be greater than and equaling in the newborn grain sum of 0.5 μm, the newborn grain being greater than 1 μm is less than 6%, does not detect the newborn grain being greater than 5 μm;

When 24 months, the newborn grain particle diameter of more than 85% is at about 0.5 μm; Be greater than and equaling in the newborn grain sum of 0.5 μm, the newborn grain being greater than 1 μm is less than 6%, does not detect the newborn grain being greater than 5 μm.

The vitamin K1 content of (3) 12 samples is respectively compared with its initial value, and all between 94.2% ~ 103.7% of initial value, such as preparation example 1 is 95.7%.

The glycerol content of (4) 12 samples is respectively compared with its initial value, and all between 93.5% ~ 107.8% of initial value, such as preparation example 4 is 99.3%.

The triglyceride content of (5) 12 samples is respectively compared with its initial value, and all between 94.5% ~ 108.4% of initial value, such as preparation example 6 is 98.4%.

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

In addition, emulsion composition shading under the room temperature of about 25 DEG C prepared by reference examples 18 preserves 24 months, value when comparing 24 months and the situation of change between 0 month initial value, and result shows:

When 0 month, the newborn grain particle diameter of more than 90% is at about 0.5 μm; Be greater than and equaling in the newborn grain sum of 0.5 μm, the newborn grain being greater than 1 μm is less than 5%, does not detect the newborn grain being greater than 5 μm;

When 24 months, the newborn grain particle diameter of less than 80% is at about 0.5 μm; Be greater than and equaling in the newborn grain sum of 0.5 μm, the newborn grain being greater than 1 μm reaches 18%, and detects many newborn grains being greater than 5 μm.

Claims (10)

1. an emulsion composition, it consists of:

Vitamin K1: 0.5-2%,

Soybean oil: 7.5 ~ 15%,

Phospholipid: 0.8 ~ 2%,

Glycerol: 1.5 ~ 3%,

PH adjusting agent: appropriate, regulates the pH value of this emulsion composition to be 7.5 ~ 8.5,

Water: appropriate, adds to 100%.

2. emulsion composition according to claim 1, wherein said pH adjusting agent is sodium hydroxide and/or hydrochloric acid.

3. emulsion composition according to claim 1, it consists of:

Vitamin K1: 0.75-1.5%,

Soybean oil: 8 ~ 12%,

Phospholipid: 1.0 ~ 1.5%,

Glycerol: 1.75 ~ 2.5%,

PH adjusting agent: appropriate,

Water: appropriate, adds to 100%.

4. emulsion composition according to claim 1, it consists of:

Vitamin K1: 0.9-1.1%,

Soybean oil: 9 ~ 11%,

Phospholipid: 1.0 ~ 1.5%,

Glycerol: 1.95 ~ 2.45%,

PH adjusting agent: appropriate,

Water: appropriate, adds to 100%.

5. emulsion composition according to claim 1, it consists of:

Vitamin K1: 1.0%,

Soybean oil: 10%,

Phospholipid: 1.2%,

Glycerol: 2.2%,

PH adjusting agent: appropriate,

Water: appropriate, adds to 100%.

6. according to the emulsion composition of any one of claim 1-5, it measures according to the wet method in Pharmacopoeia of People's Republic of China version in 2010 two annex Ⅸ E granularities and particle size distribution method the 3rd method light scattering method, and the particle diameter of the newborn grain of more than 80% is below 1.0 μm; Or be greater than at particle diameter and equal in the newborn grain sum of 0.5 μm, the newborn grain that particle diameter is greater than 1 μm is less than 10%.

7., according to the emulsion composition of any one of claim 1-5, the particle diameter of its newborn grain of more than 90% is between 0.01 ~ 1.0 μm.

8., according to the emulsion composition of any one of claim 1-5, the particle diameter of its newborn grain of more than 90% is between 0.1 ~ 1.0 μm; Further, be greater than and equal in the newborn grain sum of 0.5 μm at particle diameter, the newborn grain that particle diameter is greater than 1 μm is less than 10%.

9., according to the emulsion composition of any one of claim 1-5, it is oil-in-water type injectable emulsion.

10. prepare the method for emulsion composition described in any one of claim 1-9, it comprises the following steps:

(1) vitamin K1, soybean oil, phospholipid, glycerol is taken by recipe quantity, for subsequent use;

(2) water for injection preparation total amount being about 60-90% adds stainless cylinder of steel, adds the glycerol of recipe quantity, stirs;

(3) soybean oil of recipe quantity is added in another stainless cylinder of steel, pass into nitrogen current, add vitamin K1 and the phospholipid of recipe quantity, be stirred to each material dissolution;

(4) pass into nitrogen current, oil phase is slowly added in the aqueous phase of high-speed stirred, continue high-speed stirred in good time, obtain colostrum;

(5) regulate the pH value of this colostric fluid to prescribed limit with sodium hydroxide solution, then inject and be settled to theoretical recipe quantity with water, stir, regulate solution ph to prescribed limit with sodium hydroxide or hydrochloric acid solution if desired;

(6) under nitrogen flowing, make colostrum high pressure homogenization under 40-70MPa pressure, then under 5-25MPa pressure low pressure homogenize, cooling, then use the membrane filtration of 10 μm;

(7) filtrate fill is in ampoule bottle, fills nitrogen sealing by fusing, pressure sterilizing, to obtain final product.