CN110876719B

CN110876719B - Vitamin K1 injection and preparation method thereof

Info

Publication number: CN110876719B
Application number: CN201811042943.5A
Authority: CN
Inventors: 全东琴; 王永安; 王涛; 王亚男; 李海燕; 申辽; 张亚丹
Original assignee: Academy of Military Medical Sciences AMMS of PLA
Current assignee: Academy of Military Medical Sciences AMMS of PLA
Priority date: 2018-09-06
Filing date: 2018-09-06
Publication date: 2022-08-09
Anticipated expiration: 2038-09-06
Also published as: CN110876719A

Abstract

The invention relates to the field of medicines, and relates to a vitamin K1 injection and a preparation method thereof. In particular, the present invention relates to a micelle solution and a preparation method thereof, and also relates to an injection comprising the micelle solution and a preparation method thereof.

Description

Vitamin K1 injection and preparation method thereof

Technical Field

The invention relates to the field of medicines, in particular to the field of injection preparations.

Background

Vitamin K1 is 2-methyl-3- (3,7,11, 15-tetramethyl hexadec-2-enyl) -1, 4-naphthoquinone with molecular formula of C ₃₁ H ₄₆ O ₂ Molecular weight is 450.71, and its structural formula is shown in the following formula.

Vitamin K1 is yellow to orange yellow, transparent and viscous liquid, has odorless or almost odorless smell, is extremely unstable under illumination conditions, and is easy to decompose. The vitamin K1 is easily soluble in solvents such as chloroform, ether or vegetable oil, is slightly soluble in ethanol, is almost insoluble in water, and has a refractive index of 1.525-1.528.

Vitamin K1 is an essential substance for the liver to synthesize prothrombin, and can cause blood coagulation disorders when lacking. When human blood lacks prothrombin, the blood coagulation is retarded, and at the moment, a proper amount of vitamin K1 is supplemented to promote the liver to synthesize prothrombin, so that the effects of blood coagulation and hemostasis are achieved. The vitamin K1 is mainly used for treating vitamin K1 deficiency and thrombin hypofunction, and preventing and treating neonatal spontaneous hemorrhage, biliary fistula, obstructive jaundice and hemorrhage caused by chronic diarrhea. In addition, the vitamin K1 also has the functions of easing pain and relieving bronchospasm, and has obvious treatment effect on angina caused by visceral smooth muscle angina, bile duct spasm, intestinal spasm and the like.

Vitamin K1 is an essential substance for the preactivation of blood coagulation factors II, VII, IX, X in the human body, and has the property of an endogenous substance. The vitamin K1 is used as a hemostatic drug, and is widely applied clinically due to the advantages of remarkable curative effect, safe medication, low price and the like.

Adverse drug reaction information announced by the national food and drug administration and national adverse drug reaction detection center at 2011, 12 months and 26 days (severe anaphylaxis of vigilant vitamin K1 injection) shows that 893 adverse drug reactions of the vitamin K1 injection are caused, wherein the number of allergic drug cases caused by intravenous administration accounts for 95.3% of the total amount. The vitamin K1 injection has the main serious adverse reactions as follows: dyspnea, anaphylactoid reaction, anaphylactic shock, etc. The reason for the adverse reaction caused by the vitamin K1 injection at present may be related to the following: the vitamin K1 has poor stability, is easy to decompose when being exposed to light, and can be degraded to generate impurities when being placed for a long time to deepen the color; the vitamin K1 injection preparation adjuvant contains Tween-80, propylene glycol, etc. Wherein, the Tween-80 is a surfactant, and has larger side effect.

At present, the vitamin K1 has wide clinical application and is increasingly demanded. A new formulation with high safety and good stability is required to be developed in order to achieve the aim of improving clinical safety.

Disclosure of Invention

In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings that are commonly understood by those skilled in the art. Also, the laboratory procedures referred to herein are all conventional procedures widely used in the corresponding field. Meanwhile, in order to better understand the present invention, the definitions and explanations of related terms are provided below.

In the present invention, the term "micelle" refers to an assembly having a solid core-shell structure in which an amphiphilic substance is present in a solvent (e.g., water) at a concentration exceeding a certain critical value, as a result of mutual attraction of solphobic portions (e.g., hydrophobic portions) or solvophilic portions (e.g., hydrophilic portions), the core of which is composed of the solvophobic portions (e.g., hydrophobic portions) of the molecule, and the shell of which is composed of the solvophilic portions (e.g., hydrophilic portions) of the molecule. The micelle system can be used for wrapping/loading the medicament to realize the solubilization of the medicament.

In the invention, the term "mixed micelle" refers to a micelle system formed by compounding substances with surface activity, and can improve the solubilization performance of a single surface active substance.

In the present invention, the term "related substances" refers to impurities such as starting materials, intermediates, polymers, side reaction products, and degradation products during storage, which are introduced during the production of the pharmaceutical product. Such impurities are often referred to as related substances because their chemical structures are generally similar to or have a source relationship with the active ingredient. The term "related substances" may also refer to the content of the above-mentioned impurities in the pharmaceutical product.

In the present invention, the term "injection" refers to a sterile preparation for injection into the body, which is prepared from the raw material drug (main drug) and appropriate auxiliary materials, and includes sterile solutions, suspensions and emulsions, and sterile powders or concentrated solutions for preparing solutions, suspensions or emulsions just before use. The term "injection" refers to liquid injections.

In the present invention, the "labeled amount" refers to the specification and type of the drug, and for example, for an injection with a labeled amount of "1 mL, 10 mg", the labeled amount means that 1mL of the injection contains 10mg of the principal drug. The content of the injection is the percentage of the actual amount of the main drug in the injection to the marked amount, and can be used for evaluating the chemical stability of the injection.

In the present invention, the term "egg yolk lecithin E80" means egg yolk lecithin having a purity of not less than 80%, which generally means phosphatidylcholine purity by TLC-FID detection.

The invention aims to solve the technical problem of providing a vitamin K1 injection which can be produced in an enlarged scale and can resist high-temperature sterilization. The present inventors prepared an injection comprising vitamin K1 mixed micelles using a Bile salt/phospholipid mixed micelles system (BS/PC-MMS) by using a specific formulation and method, which has good physical and chemical stability against high temperature sterilization, and can improve the safety of drugs. The following invention is thus provided:

in one aspect, the present application provides a micelle solution comprising water and a mixed micelle consisting of vitamin K1, sodium glycocholate and a phospholipid, wherein the micelle solution has a vitamin K1 weight percentage of (1.00 ± 0.05%), sodium glycocholate weight percentage of (5.5 ± 0.1%) to (6.0 ± 0.1%), and phospholipid weight percentage of (6.0 ± 0.1%).

In certain embodiments, the phospholipid is a lecithin, such as egg yolk lecithin (e.g., egg yolk lecithin E80).

In certain embodiments, the weight percent of sodium glycocholate is (5.5 ± 0.1)%, (5.6 ± 0.1)%, (5.7 ± 0.1)%, (5.8 ± 0.1)%, (5.9 ± 0.1)%, or (6.0 ± 0.1)%.

In certain embodiments, the water is water for injection.

Optionally, the micellar solution further comprises a pH adjusting agent (e.g. hydrochloric acid or sodium hydroxide).

In certain embodiments, the mixed micelle is a spheroidal entity.

In certain embodiments, the pH of the micellar solution is 3.5 to 7.0, such as 3.5 to 4.0, 4.0 to 4.5, 4.5 to 5.0, 5.0 to 6.0, 6.0 to 6.5, or 6.5 to 7.0.

In certain embodiments, the mixed micelles have an average particle size of 5 to 30nm, such as 5 to 15nm, 15 to 20nm, or 20 to 30nm, such as 9.7 ± 2.0 nm.

In certain embodiments, the Zeta potential of the mixed micelle is between-40 mV and-50 mV.

In certain embodiments, the micellar solution has a light transmittance of greater than 90%.

In one aspect, the present application provides a method of preparing a micellar solution, comprising the steps of:

step 1: mixing vitamin Kl, sodium glycocholate, phospholipid and a proper amount of ethanol, and stirring at 40 +/-2 ℃ in a dark place until the solution is clear; wherein the weight ratio of the vitamin Kl, the sodium glycocholate and the phospholipid is (1.00 +/-0.05): (5.5. + -. 0.1) - (6.0. + -. 0.1): (6.0 +/-0.1);

and 2, step: introducing inert gas into the solution obtained in the step (1) at 40 +/-2 ℃ in a dark condition, and volatilizing to remove ethanol to obtain a semi-solid mixture;

and step 3: mixing the semi-solid mixture obtained in the step (2) with water, stirring, and adjusting the pH to 5.3-6.0 by using a pH regulator to completely dissolve the system until the system is clear; adding water again to fix the volume; the ratio of the total volume of water added (mL) to the weight of vitamin K1 in solution (g) was 100: (1.00. + -. 0.05).

In certain embodiments, the phospholipid in step 1 is a lecithin, such as egg yolk lecithin (e.g., egg yolk lecithin E80).

In certain embodiments, the ratio of the volume of ethanol in step 1 to the total volume of water added in step 3 (mL/mL) is 1-5: 100.

In certain embodiments, the inert gas in step 2 is nitrogen.

In some embodiments, the inert gas is introduced in the step 2 for 1 to 2 hours.

In certain embodiments, the water in step 3 is water for injection.

In certain embodiments, in step 3, the water added prior to adjusting the pH comprises 70% to 90% (e.g., 80%) of the total amount of water added.

In certain embodiments, the pH adjusting agent added in step 3 is selected from hydrochloric acid and sodium hydroxide.

In certain embodiments, in step 3, the pH is adjusted to 5.4.

In certain embodiments, the micellar solution produced by the method is a micellar solution as described above.

In one aspect, the present application also provides an injection comprising the micellar solution of the invention and a package (e.g. an ampoule).

In one aspect, the present application also provides a method of preparing an injection as described above, comprising the steps of:

step 1: providing a micellar solution of the invention;

step 2: the micellar solution of the present invention is filtered, filled, sealed and sterilized.

In certain embodiments, the filling refers to filling the micellar solution into ampoules.

In certain embodiments, the sealing is a melt seal.

In certain embodiments, the sterilization refers to high temperature sterilization (e.g., sterilization at 121 ℃ for 15 min).

In one aspect, the present application provides the use of the micellar solution or injection as described above for the prevention and/or treatment of vitamin K1 deficiency, thrombasthenia, neonatal spontaneous hemorrhage, hemorrhagic symptoms due to biliary fistula, obstructive jaundice or chronic diarrhea, visceral smooth muscle angina, bile duct spasm or cramp caused by intestinal spasm in a patient.

The injection of the present invention may be administered to a patient in any suitable manner of administration, for example, subcutaneous injection, intramuscular injection, intravenous bolus injection or intravenous drip. In certain preferred embodiments, the injection is administered to the patient in a prophylactically or therapeutically effective amount. It is well within the ability of those skilled in the art to determine such effective amounts. For example, an amount effective for therapeutic use will depend on the severity of the disease to be treated, the general state of the patient's own immune system, the general condition of the patient, e.g., age, weight and sex, the mode of administration of the drug, and other treatments administered concurrently, and the like.

The invention adopts a bile salt/phospholipid mixed micelle system to prepare the vitamin K1 injection, which not only can obviously improve the solubility of insoluble vitamin K1, but also ensures the safety of the preparation because the phospholipid and the bile salt contained in the micelle have biocompatibility and are called as physiological excipients. Although the prior art adopts a freeze-drying re-dissolution method or a film dispersion-ultrasonic method to prepare a mixed micelle system of bile salt/phospholipid, however, the freeze-drying reconstitution method requires the use of an organic solvent such as 2-methyl-2-propanol or 2-methyl-2-butanol, the vitamin K1 is oily substance, the phenomenon of liquefaction caused by combination of oil drops and dissolution of cholate phospholipid is easily generated after water is removed, the oil-water separation phenomenon is caused when water is added for redissolution, the whole preparation is difficult to recover to a uniform clear solution before freeze-drying, so the problems in the aspects of solvent residue, safety, environmental protection and the like easily occur, and the freeze-drying process needs to be carried out under the condition of vacuum low temperature (-40 ℃ to-25 ℃), the membrane dispersion-ultrasonic method requires reduced pressure distillation, and both methods have higher requirements on equipment and processes required by industrial scale-up production. Compared with the prior art, the preparation method can be implemented under normal pressure, has simple process and is more suitable for industrial scale-up production.

In the preparation method of the invention, the concentration of the bile salt and the phospholipid has important influence on physical stability indexes such as light transmittance, average particle size and the like of the vitamin Kl mixed micelle system (see figures 1-2), and is an important factor influencing the molding and stability of the sterilized injection. The inventor finds that when the concentrations of phospholipid and cholate are lower, the drug loading rate of the mixed micelle is lower, the particle size of the formed mixed micelle system is larger, the solution is cloudy, and the light transmittance is lower; however, when the concentrations of phospholipid and bile salt are high, the amount of ethanol required for preparation is large, and the prepared solution has high viscosity, which brings inconvenience and safety risks to clinical medication.

In the prior art, the drug loading of a micelle preparation of the same kind is 5mg/mL, but the inventor of the invention finds that the drug loading of the micelle preparation can reach 10mg/mL under a proper phospholipid concentration, and particularly finds that when the phospholipid concentration is lower than 6%, the molding and stability of an injection are poor, and when the concentration of phospholipid or cholate is too high, the molding and stability of the injection are not obviously changed, which indicates that the preparation of the mixed micelle injection is not significant when the concentration of phospholipid or cholate is too high.

In addition, the type of bile salt has an important influence on the pH range of the micellar solution, the chemical stability of high temperature sterilization (as indicated by content, impurities and appearance) (see fig. 3 to 6). When the type of bile salt is changed from sodium glycocholate to sodium deoxycholate, the lowest pH value for maintaining the physical stability of the mixed micelle system is increased from 3.5 to 8.5 (see figures 3 to 4), the risk of chemical degradation in the high-temperature sterilization process is increased, the content is obviously reduced, the related substances are obviously increased (see figure 5), and the appearance of the solution is obviously changed (see figure 6).

The inventor finds that when the composition of the micelle solution is 11.0 percent of vitamin K, 6.0 percent of sodium glycocholate and 6.0 percent of egg yolk lecithin, and the pH value of the solution is adjusted to be 5.4, the average particle size of the micelle in the prepared micelle solution is 9.7 +/-2.0 nm, the light transmittance of the solution is more than 90 percent, the content of the main drug is 98.0 +/-2.0 percent of the marked amount, the content is not reduced after sterilization, and the impurities are not increased.

Advantageous effects

Compared with the prior art, the technical scheme of the invention has the following beneficial effects:

1. the vitamin Kl mixed micelle solution prepared by the method has good physical and chemical stability of tolerating high-temperature sterilization, and can improve the safety of clinical use when being used as an injection;

2. the preparation method of the invention has the advantages of higher safety, simple process, low preparation cost, good process reproducibility, no need of special equipment and easy industrialized mass production.

Drawings

FIG. 1 shows the effect of bile salt concentration or phospholipid concentration on the light transmittance of micellar solution studied in a single variable manner in example 4. FIG. 1A shows the transmittance of micellar solutions at different bile salt concentrations (2% to 7%); FIG. 1B shows the light transmittance of micellar solutions at different phospholipid concentrations (4% to 7%). As shown in the figure, when the concentrations of phospholipid and cholate are lower, the formed mixed micelle solution is cloudy, and the light transmittance is lower; when the phospholipid concentration reached about 6.0%, the light transmittance of the solution did not change significantly.

FIG. 2 shows the effect of bile salt concentration or phospholipid concentration on micelle size studied in a single variable manner in example 4. FIG. 2A shows the average particle size of micelles at different phospholipid concentrations (4% to 7%); FIG. 2B shows the average particle size of micelles at different bile salt concentrations (4% to 7%). As shown in the figure, when the concentrations of phospholipid and bile salt are low, the particle size of the formed mixed micelle is larger; when the phospholipid concentration reached about 6.0%, there was no significant change in micelle size.

FIG. 3 shows the change of light transmittance of micellar solutions with changes in pH of the solutions in example 5, using sodium glycocholate and sodium deoxycholate, respectively, as bile salts.

FIG. 4 shows the changes in the average particle size (FIG. 4A) and Zeta potential (FIG. 4B) of micelles with changes in the pH of the solution in example 5 using sodium glycocholate and sodium deoxycholate, respectively, as the cholate.

As shown in fig. 3 and 4, the lowest pH for maintaining physical stability of the mixed micelle system was 3.5 using sodium glycocholate as the bile salt; and sodium deoxycholate is used as bile salt instead, and the lowest pH value for maintaining the physical stability of the mixed micelle system is increased to 8.5.

FIG. 5 shows the contents of sodium deoxycholate (FIG. 5A) and sodium glycocholate (FIG. 5B) before and after sterilization in the injection prepared in example 5 with cholate as the cholate and the related substances. As shown in the figure, sodium deoxycholate is used as cholate, so that the content is obviously reduced after sterilization, and related substances are obviously increased; and the content and related substances of the sodium glycocholate used as the cholate do not obviously change before and after sterilization.

Fig. 6 shows the shape of the injection before and after sterilization at various concentrations when sodium glycocholate and sodium deoxycholate were used as the cholate components, respectively, and the solution color before and after sterilization showed strong stability using the sodium glycocholate injection, while the sodium deoxycholate component produced a significant color change after sterilization, indicating that the sodium glycocholate component was significantly superior to the sodium deoxycholate component.

Fig. 7 shows the morphology of the mixed micelle prepared in example 1 under TEM, and as shown, the mixed micelle is a uniformly distributed spheroidal entity.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer.

EXAMPLE 1 preparation of vitamin K1 Mixed micelle solution and injection

The specification of the injection to be prepared in this embodiment is 1 mL/injection, the amount of vitamin K1 containing the main drug per injection is 10mg, and the composition is as follows, calculated by 1000 injections:

the specific operation process is as follows:

(1) weighing the components according to the proportion, placing the components in a beaker, adding a proper amount of ethanol, placing the mixture in a water bath magnetic stirrer, heating to 40 +/-2 ℃, stirring, and dissolving until the mixture is clear;

(2) preserving the temperature of the solution obtained in the step (1) at 40 +/-2 ℃, keeping the solution away from light, introducing nitrogen for 1-2 h, and volatilizing to remove ethanol to obtain a semi-solid mixture;

(3) and (3) adding 80% of the prescription amount of water for injection into the semisolid mixture obtained in the step (2), adjusting the pH to 5.4, continuously stirring until the mixture is clear, and supplementing the water for injection to the prescription amount (the pH is not changed greatly in actual measurement) to obtain a solution containing vitamin K1 mixed micelles.

(4) And (4) filtering the solution obtained in the step (3), filling the filtered solution into an ampoule, sealing the ampoule by melting, and sterilizing the ampoule at 121 ℃ for 15min to obtain the injection.

Example 2 physicochemical Properties of micellar solution

(1) Particle diameter and Zeta potential measurement

The solution containing vitamin K1 mixed micelles from example 1 was diluted to 0.5mg/mL and the mean particle size was determined using a Malvern Nano ZS90Zetasizer to give micelles with a mean particle size of 9.7. + -. 2.0 nm.

(2) Morphological investigation

And (3) dropwise adding the diluted mixed micelle solution to a copper mesh, counterdyeing with 2% sodium phosphotungstate solution, observing under a Transmission Electron Microscope (TEM) and taking a picture. The prepared mixed micelle is a uniformly distributed sphere-like entity (as shown in figure 7).

(3) Transmittance test of solution

The mixed micelle solution is taken, and the light transmittance (%) of the mixed micelle solution is measured by an ultraviolet-visible spectrophotometer under 600 nm. The transmittance of the solution was measured to be 94.7%.

(4) Content and related substance determination

(4-1) content determination: measured according to high performance liquid chromatography (appendix XE of the second part of the Chinese pharmacopoeia 2010 edition).

Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filler, and water-methanol (40:60) is used as a mobile phase; the detection wavelength was 270 nm.

The determination method comprises the following steps: precisely weighing a proper amount of a sample to be detected, adding a mobile phase for dissolving, quantitatively diluting into a solution containing 0.1mg in each 1mL, precisely measuring 20 mu L, injecting into a liquid chromatograph, and recording a chromatogram; and taking a proper amount of vitamin K1 reference substances, and measuring by the same method. Calculating according to the peak area by an external standard method to obtain the product.

The content of the injection is expressed by the percentage of the actual content in the marked amount, and the calculation formula is as follows:

(4-2) measurement of related substances: taking a sample to be tested, adding a mobile phase for dissolving and diluting to prepare a solution containing 1mg per 1mL as a test solution; precisely measuring 1mL, and placing in a 100mL measuring flask to serve as a control solution; according to the chromatographic conditions under the content determination item, precisely measuring 20 mu L of each of the test solution and the control solution, respectively injecting the test solution and the control solution into a liquid chromatograph, recording the chromatogram until the retention time of the main component peak is 2 times that of the chromatogram, wherein if an impurity peak exists in the chromatogram of the test solution, the peak area of a single impurity is not more than 0.5 times (0.5%) of the main peak area of the control solution, and the sum of the peak areas of the impurities is not more than 2.0 times (2.0%) of the main peak area of the control solution.

(4-3) measurement results: the content is 98.3 percent and the content of related substances is 0.57 percent.

Example 3 2-year stability study of injection

Sterile injections were prepared according to the method of example 1 and subjected to a 2-year stability study, with the results shown in table 1.

TABLE 1 stability data for vitamin K1 phospholipid/bile salt mixed micelle sterile injectable solutions

As can be seen from Table 1, the injection of the present invention has good stability and can be stored for more than 2 years.

Example 4 investigation of the Effect of bile salt and phospholipid concentrations on the Properties of micellar solutions

Vitamin K1 mixed micelle solution was prepared according to the method of example 1, and the influence of the bile salt concentration and the phospholipid concentration on the solution properties was investigated in a single variable manner.

FIG. 1A shows the transmittance of micellar solutions at different bile salt concentrations (2% to 7%); FIG. 1B shows the light transmittance of micellar solutions at different phospholipid concentrations (4% to 7%). FIG. 2A shows the average particle size of micelles at different phospholipid concentrations (4% to 7%); FIG. 2B shows the average particle size of micelles at different bile salt concentrations (4% to 7%). In fig. 1A and 2B, the conditions other than the bile salt concentration were the same as in example 1. In FIGS. 1B and 2A, the conditions other than the phospholipid concentration were the same as in example 1.

As shown in the figure, when the concentrations of phospholipid and cholate are lower, the particle size of the formed mixed micelle system is larger, the solution is cloudy, and the light transmittance is lower; when the phospholipid concentration reached about 6.0%, there was no significant change in the light transmittance of the solution or in the particle size of the micelles. If the concentrations of phospholipid and bile salt are high, the amount of ethanol required in preparation is large, and the prepared solution has high viscosity, so that inconvenience and safety risks are easily brought to clinical medication. It was found that a solution having a relatively good light transmittance was obtained even when the bile salt concentration was about 6.0%, the phospholipid concentration was about 6.0%, and the bile salt concentration was about 5.5%.

Example 5 investigation of the Effect of the type of bile salt on the stability of micellar solutions

Vitamin K1 mixed micellar solution was prepared according to the method of example 1 and the influence of the type of bile salts on the pH tolerated by the micellar solution was investigated in a univariate manner.

Figures 3 and 4 show the pH at which the physical stability of the mixed micellar system is maintained using sodium glycocholate and sodium deoxycholate, respectively, as bile salts. Fig. 3 shows the change of light transmittance of the micelle solution according to the change of the pH of the solution, fig. 4A shows the change of the average particle size of the micelle according to the change of the pH of the solution, and fig. 4B shows the change of Zeta potential of the micelle according to the change of the pH of the solution. As shown, the lowest pH for maintaining the physical stability of the mixed micelle system was 3.5 using sodium glycocholate as the bile salt, whereas the lowest pH for maintaining the physical stability of the mixed micelle system was raised to 8.5 using sodium deoxycholate as the bile salt instead.

Too high a pH of the injection increases the risk of chemical degradation of the drug during high temperature sterilization. The contents of the injection solution and the related substances were measured according to the method of example 2, and the results are shown in fig. 5. Fig. 5A and 5B show the content of sodium deoxycholate and sodium glycocholate before and after sterilization, respectively, of the injection prepared using sodium deoxycholate and sodium glycocholate as bile salts, and the related substances. As shown in the figure, sodium deoxycholate is used as cholate, so that the content is obviously reduced after sterilization, and related substances are obviously increased; and the content and related substances of the sodium glycocholate used as the cholate do not obviously change before and after sterilization.

In addition, the pH of the injection solution is too high or too low, which easily causes adverse effects on the human body. The pH of the injection solution is usually controlled to be between 4 and 9. As shown in FIGS. 3 and 4, the micellar solution of the present invention can be stable at a pH in the range of 3.5 to 7 and is suitable for use as an injection.

FIG. 6 shows the shape of the injection before and after sterilization at each concentration when sodium glycocholate (A) and sodium deoxycholate (B) were used as the cholate component, respectively. The samples in fig. 6A were sodium deoxycholate as bile salts, the four samples on the left were pre-sterilized injections, and the bile salt concentrations were 4%, 5%, 6%, and 7% in this order, and the four samples on the right were sterilized injections, and the bile salt concentrations were 4%, 5%, 6%, and 7% in this order. The samples in fig. 6B were sodium glycocholate as bile salt, the two samples on the left were pre-sterilized injections, and the bile salt concentration was 6%, and the two samples on the right were sterilized injections, and the bile salt concentration was 6%. As shown in the figure, the color of the solution is basically unchanged before and after sterilization by using various groups of injection of sodium glycocholate, and the solution shows stronger stability, while the injection of the sodium deoxycholate group generates obvious color change after sterilization, which shows that the sodium glycocholate group is obviously superior to the sodium deoxycholate group.

While specific embodiments of the invention have been described in detail, those skilled in the art will understand that: various modifications and changes in detail can be made in light of the overall teachings of the disclosure, and such changes are intended to be within the scope of the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims

1. A micellar solution comprises water and a mixed micelle consisting of vitamin K1, sodium glycocholate and egg yolk lecithin, wherein the micellar solution contains (1.00 +/-0.05%) by weight of vitamin K1, (5.5 +/-0.1)% to (6.0 +/-0.1%) by weight of the sodium glycocholate and (6.0 +/-0.1)% by weight of the egg yolk lecithin; the micellar solution is prepared by a process comprising the steps of:

step 1: mixing vitamin Kl, sodium glycocholate, egg yolk lecithin and appropriate amount of ethanol, and stirring at 40 + -2 deg.C in the dark until the solution is clear; wherein the weight ratio of the vitamin Kl, the sodium glycocholate and the egg yolk lecithin is (1.00 +/-0.05): (5.5. + -. 0.1) - (6.0. + -. 0.1): (6.0 +/-0.1);

step 2: introducing inert gas into the solution obtained in the step (1) at 40 +/-2 ℃ in a dark condition, and volatilizing to remove ethanol to obtain a semi-solid mixture;

2. The micellar solution of claim 1, wherein said water is water for injection.

3. The micellar solution of claim 1, further comprising a pH adjusting agent.

4. The micellar solution of claim 3, wherein said pH adjusting agent is hydrochloric acid or sodium hydroxide.

5. The micellar solution of any one of claims 1 to 4, having one or more of the following characteristics:

(1) the mixed micelle is a sphere-like entity;

(2) the pH value of the micelle solution is 3.5-7.0;

(3) the average particle size of the mixed micelle is 5-30 nm;

(4) the Zeta potential of the mixed micelle is-40 mV to-50 mV;

(5) the light transmittance of the micellar solution is greater than 90%.

6. A method of preparing a micellar solution, comprising the steps of:

step 1: mixing the vitamin Kl, sodium glycocholate, egg yolk lecithin and a proper amount of ethanol, and stirring at 40 +/-2 ℃ in a dark place until the solution is clear; wherein the weight ratio of the vitamin Kl, the sodium glycocholate and the phospholipid is (1.00 +/-0.05): (5.5. + -. 0.1) - (6.0. + -. 0.1): (6.0 +/-0.1);

7. The method of claim 6, having one or more of the following features:

(1) the ratio (mL/mL) of the volume of the ethanol in the step 1 to the total volume of the water added in the step 3 is 1-5: 100;

(2) the inert gas in the step 2 is nitrogen;

(3) in the step 2, the time for introducing the inert gas is 1-2 hours;

(4) the water in the step 3 is water for injection;

(5) in step 3, the water added before the pH is adjusted accounts for 70-90% of the total amount of the added water;

(6) the pH regulator added in step 3 is selected from hydrochloric acid and sodium hydroxide.

8. An injection comprising the micellar solution of any one of claims 1 to 5 and a package.

9. The injection of claim 8, wherein the package is an ampoule.

10. A process for the preparation of an injection according to claim 8 or 9, comprising the steps of:

step 1: providing the micellar solution of any one of claims 1 to 5;

step 2: filtering, filling, sealing and sterilizing the micellar solution of any one of claims 1 to 5.

11. Use of the micellar solution according to any one of claims 1 to 5 or the injectable formulation according to claim 8 or 9 for the preparation of a medicament for the prevention and/or treatment of vitamin K1 deficiency, hypothrombin, spontaneous neonatal hemorrhage, biliary fistula, bleeding symptoms caused by obstructive jaundice or chronic diarrhea, visceral smooth muscle colic, colic caused by bile duct spasm or intestinal spasm in a patient.