CN111358754A - Long-circulating emulsion of macrolide antibiotics and preparation method thereof - Google Patents
Long-circulating emulsion of macrolide antibiotics and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of food, and discloses a macrolide antibiotic long-circulating emulsion and a preparation method thereof, wherein the macrolide antibiotic long-circulating emulsion comprises the following components in percentage by mass: 0.1-1% of macrolide antibiotics, 10-30% of oil phase solvent, 0.5-5% of emulsifier, 0.2-5% of co-emulsifier, 0.3-5% of potential regulator, 1-5% of osmotic pressure regulator and the balance of water for injection; the emulsifier comprises a pegylated synthetic phospholipid. The invention overcomes the problems of adverse reaction and drug resistance brought by the traditional macrolide antibiotic administration route, and simultaneously enhances the antibacterial effect of the macrolide antibiotic in partial strains.
Description
Technical Field
The invention relates to the technical field of pharmaceutics, in particular to a macrolide antibiotic long-circulating emulsion and a preparation method thereof.
Background
The macrolide drug is a weakly basic antibiotic produced by streptomyces, contains a 14-or 16-membered ring with a lactone structure, and has strong inhibiting effect on streptococcus pneumoniae. The main pharmacological action mechanism is that the antibacterial peptide acts on a bacterial ribosome 50S subunit, blocks the synthesis of mycoprotein and the displacement of messenger ribonucleic acid, and has better antibacterial action on atypical pathogenic bacteria such as chlamydia, mycoplasma, legionella, helicobacter pylori and the like. Currently, macrolide antibiotic preparations on the market are granules, dispersible tablets, sustained-release tablets and dry suspensions. In addition, the macrolide antibiotics have poor solubility and low absorption utilization rate, so that the clinical dosage of the tablets is high, such as 250mg and 500mg of clarithromycin, 500mg of azithromycin, 500mg of dirithromycin, 200mg of rotamycin and the like.
The traditional antibiotic oral therapy in the administration mode has the problems of rapid metabolism in ① body, less reaching of infected parts, low effective utilization rate, ② large dose, long-period administration, serious adverse reaction, ③ difficulty in realizing intracellular enrichment and incapability of killing intracellular living bacteria, ④ generation of antibiotic resistance and the like.
Although some studies have been made on the administration of antibiotics as injections, macrolide antibiotics generally have poor water solubility and are difficult to directly prepare as injections. Clinically, the macrolide antibiotics are prepared into injection, and ethanol or cosolvents such as malic acid and lactic acid are usually added to increase the solubility of the macrolide antibiotics in an aqueous solution. Typically, ethanol is widely used as a cosolvent commonly used in clinic, and the property of ethanol solves the problem that many drugs cannot be clinically used due to poor solubility. But still has a plurality of adverse reactions such as phlebitis, injection site pain and other clinical compliance problems, and has great replaceability. Other surfactants such as malic acid, lactic acid, propylene glycol, mannitol and Tween series can increase the solubility of the medicament to a certain extent, but obvious adverse reactions are found in clinical application. The addition of cosolvents such as ethanol and the like can easily cause more adverse reactions in clinic, so that the traditional macrolide antibiotic injection is not commonly used in clinic. It is desirable to develop a formulation that improves the compliance of clinical patients while achieving therapeutic goals.
With the widespread use and abuse of antibiotics, the problem of antibiotic resistance has become a global public health safety issue. Emulsions have been widely used clinically as technically mature nano delivery systems. However, no research report on antibiotic injection emulsion exists at present, which is mainly because antibiotic drugs mostly belong to macrolide substances, and the antibiotic drugs have extremely poor lipid solubility and are insoluble in a lipid-soluble solvent, so that the antibiotic drugs cannot be prepared into emulsion. Therefore, the biggest difficulty in how to use an emulsion as a drug delivery system is to solve the problem of solubility in the oil phase.
Disclosure of Invention
The invention aims to overcome the problems of ① rapid in vivo metabolism, low effective utilization rate, ② large dose, long-period administration, serious adverse reaction, ③ difficulty in realizing intracellular enrichment, incapability of killing intracellular living bacteria, ④ generation of antibiotic resistance and the like caused by the traditional administration mode of macrolide antibiotics.
In order to solve the technical problems, the invention adopts the following technical scheme:
a macrolide antibiotic long-circulating emulsion comprises the following components in percentage by mass: 0.1-1% of macrolide antibiotics, 10-30% of oil phase solvent, 0.5-5% of emulsifier, 0.2-5% of co-emulsifier, 0.3-5% of potential regulator, 1-5% of osmotic pressure regulator and the balance of water for injection; the structure of the macrolide antibiotic comprises-NH2One or more of a group, -OH group; before use, the macrolide antibiotics are esterified to form macrolide antibiotic fatty acid esters.
The macrolide antibiotics mostly belong to BCS IV drugs, have the characteristics of low solubility and low permeability, can promote transmembrane absorption of the drugs by virtue of phagocytosis of cells and exchange action of cell membranes and external phospholipids through preparing the macrolide antibiotics into a fat emulsion preparation, and improve the permeability of the drugs so as to improve the bioavailability of the drugs. However, macrolides are very poorly lipid soluble and insoluble in lipid soluble solvents, making it very difficult to prepare them into emulsions. Macrolide antibiotic molecules suitable for use in the present invention have at least one-NH in their structure2The group or one-OH group is used as a base, and is esterified to form macrolide antibiotic fatty acid ester which can be dissolved in an oil phase solution, so that the drug loading is improved, and the emulsion auxiliary drug delivery is prepared.
The emulsion is emulsion droplet particles formed by water phase, oil phase and emulsifier through specific preparation process and wrapped by emulsion film, wherein the inside of the emulsion film is drug-loaded oil phase, and the outside of the emulsion film is water phase. The special physicochemical property of the emulsion membrane enables the emulsion membrane to be better fused with a biological membrane so as to realize the absorption of the cells to emulsion granules, after the emulsion membrane is absorbed into the cells, the emulsion membrane is broken by enzymolysis to release drugs, thereby realizing the killing of the living bacteria in the cells, overcoming the problem that the traditional administration mode of antibiotics can not penetrate through the cell membrane to enter the cells, and realizing the beneficial effect of effectively inhibiting chronic infection and infection recurrence. In addition, the antibiotic is prepared into the nano emulsion for intravenous injection, so that the problems of incomplete absorption of oral administration drugs and adverse reactions caused by ethanol and other irritant cosolvents existing in the conventional injection administration are solved, and the blood concentration level of the conventional administration mode of the antibiotic can be achieved by using lower dosage. The technical scheme of the invention adopts a method of esterifying antibiotics and fatty acid to improve the polarity of the antibiotics and increase the lipid solubility of the antibiotics. By preparing the antibiotic medicines into the emulsion for injection, the first-pass effect of the medicines can be avoided firstly, the medicines directly enter blood by veins, the medicine dosage can be greatly reduced, and the steady blood concentration can be reached.
Preferably, the emulsifier comprises a pegylated synthetic phospholipid.
The PEG synthetic phospholipid is added into the prescription composition of the traditional emulsion, and the PEG chain is wrapped outside the emulsion membrane, so that the PEG synthetic phospholipid can avoid the cleaning effect (phagocytosis of MPS) of the reticuloendothelial system in vivo, can stably exist in a circulatory system, prolongs the half life and maintains the steady blood concentration for a longer time.
The PEGylated synthetic phospholipid refers to PEGylation of synthetic phospholipid, such as DSPE-PEGylated phospholipid, DPPE-PEGylated phospholipid, etc. In addition, the PEG can be classified into PEGylated phospholipids with different molecular weights, such as DSPE-PEG2000, DSPE-PEG5000, etc., according to the quantity of the connected PEG. Studies have shown that different pegylated synthetic phospholipids can act to avoid MPS phagocytosis. While the longer the length of the PEG chain, the thicker the steric hindrance layer it forms on the surface of the emulsion droplet, the thicker the steric hindrance layer is, but the better it is not. PEG with the molecular weight of 1500-6000, particularly PEG with the molecular weight of 2000-5000 is preferably used, so that the long circulation effect can be ensured.
Preferably, the esterification treatment is performed by: and (3) carrying out esterification reaction with the macrolide antibiotic by using saturated fatty acid to form the macrolide antibiotic fatty acid ester.
Preferably, the carbon chain length of the saturated fatty acid is 6-18.
Preferably, the saturated fatty acid comprises one of caproic acid, heptanoic acid, caprylic acid, pelargonic acid, capric acid, lauric acid, myristic acid, palmitic acid and stearic acid.
Macrolide antibiotic molecules suitable for use in the present invention have at least one-NH in their structure2The group or one-OH group is poor in lipid solubility, and the fatty acid macrolide antibiotic which can be dissolved in an oil phase solution and is obtained by esterification reaction with saturated fatty acid with 6-18 carbon atoms is used for improving the drug loading rate. The carbon chain of the saturated fatty acid is too short, and the esterified molecule has larger polarity, so that the saturated fatty acid is difficult to prepare into emulsion; the carbon chain is too long, the esterified molecules are too large, and the transmembrane and fusion of cells are difficult to complete even if the esterified molecules are prepared into an emulsion, and the esterified molecules are difficult to penetrate cell membranes and enter the cells, so that a good administration effect cannot be achieved. Through intensive research and experiments, the saturated fatty acid with 6-18 carbon atoms is finally determined as the optimal choice, and the good absorption effect can be kept while the milk forming property is ensured.
Preferably, the potential modifier is a positive charge modifier.
The positive charge potential regulator is added in the prescription, so that the emulsion droplet particles are positively charged, the characteristic of negative charge of a bacterial microenvironment is utilized, passive targeting is realized, the enrichment concentration of the medicament at the infected part is increased, and the antibacterial effect is improved. And the advantage of good compatibility of the emulsion membrane and the biological membrane is favorable for emulsion droplet particles to penetrate through the biological membrane and enter the interior of cells. The average particle size of the emulsion is between 100 and 500nm, and the Zeta potential is between 10 and 50 mV.
Preferably, the macrolide antibiotic is one of azithromycin, clarithromycin, roxithromycin, dirithromycin, fluoroerythromycin and rotamycin; the oil phase solvent comprises one or more of fatty acid triglyceride; the emulsifier also comprises one or more of egg yolk phospholipid, soybean phospholipid, phosphatidylcholine and 15-hydroxystearic acid polyethylene glycol ester; the auxiliary emulsifier comprises one or more of polysorbate 80, poloxamer 188, sodium dodecyl sulfate and hexadecyl ammonium bromide; the potential regulator comprises one or more of octadecylamine, DOTAP and cationic surfactant; the osmotic pressure regulator comprises one or more of glycerol and mannitol.
The azithromycin, the clarithromycin, the roxithromycin, the dirithromycin, the fluoromycin and the rotamycin have at least one-NH2Macrolide antibiotics, either a group or one-OH group, can be used as drug substance in this application after esterification. However, it will be appreciated by those skilled in the art that the drug substances useful in the present application are not limited to the above-listed ones, and others may contain at least one-NH group2The macrolide antibiotics with groups or one-OH group can also be used as the raw material medicaments of the application after esterification.
Preferably, the oil phase solvent comprises one or more of soybean oil, olive oil, tea seed oil, fish oil, castor oil, coix seed oil, tricaprylin, tricaprin, caprylic capric triglyceride and trilaurin.
A preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
A. adding the macrolide antibiotics and the emulsifier into the oil phase solvent according to the proportion, continuously heating and stirring until the macrolide antibiotics and the emulsifier are completely dissolved to form a drug-loaded oil phase;
B. mixing the potential regulator, the osmotic pressure regulator and the co-emulsifier with the water according to the prescription composition, and heating until the mixture is completely dissolved to form a water phase;
C. slowly adding the drug-loaded oil phase into the water phase, shearing at a high speed under stirring, and adding the rest water to prepare primary emulsion;
D. homogenizing the primary emulsion under high pressure to obtain final emulsion;
E. and (3) sterilizing the final emulsion, filling nitrogen and sealing to obtain the macrolide antibiotic long-circulating emulsion.
In the preparation method, the adding sequence of the auxiliary materials in the preparation process of the oil phase can be the step A, or the adding sequence of the auxiliary materials can be adjusted according to the experimental requirements, but the final state of the oil phase ensures that all the components are completely dissolved and the oil phase is clear; in the preparation process of the water phase, the adding sequence of the auxiliary materials can be the step B, or the adding sequence of the auxiliary materials can be adjusted according to the experimental requirements, and finally, the state of the water phase should ensure that all the components can be completely dissolved or uniformly dispersed.
Preferably, in the step A, the heating temperature is 50-80 ℃; in the step B, the heating temperature is 50-80 ℃; in the step C, the stirring speed of high-speed shearing is 3000-20000 rpm, the shearing time is 5-25 min, and the shearing temperature is 50-80 ℃; in the step D, the high-pressure homogenization specifically comprises the following steps: controlling the homogenizing temperature to be 50-70 ℃, and circulating for 4-10 times under the pressure of 200-1500 bar; in the step E, the sterilization comprises one of filtration sterilization or high-temperature sterilization.
The final milk sterilization operation can adopt 0.22 μm filtration sterilization, filtration sterilization and filtration sterilization, and can also adopt high temperature sterilization at 121 ℃.
Compared with the prior art, the implementation of the invention has the following beneficial effects:
according to the invention, macrolide antibiotics are prepared into the nanoemulsion, and the synthesized phospholipid subjected to PEGylation is added into the formula of the emulsion, so that emulsion particles can avoid phagocytosis of reticuloendothelial cells in a human body, the effective drug concentration of the macrolide antibiotics in-vivo circulation is ensured, the in-vivo circulation metabolism time of the macrolide antibiotics nanoemulsion can be remarkably prolonged, and the precondition guarantee is provided for the antibiotics to reach more infected parts. By adding the positive charge potential regulator into the emulsion, the passive targeting between the emulsion and a bacterial microenvironment with negative charge can be realized, the enrichment concentration of the drugs at the infected part is increased, the drug concentration at the focus part is improved, the effective utilization rate of antibiotics is improved, and thus the clinical dosage of the antibiotics is reduced. Meanwhile, due to the particularity of the emulsion carrier, the emulsion carrier is similar to the components of cell membranes of human bodies, and has higher biocompatibility. The emulsion membrane formed by the phospholipid material can easily reach a focus part through various biological membranes of a human body, so that the medicament can better penetrate through the biological membranes to reach the inside of cells, and the bacteria living in the cells are further killed.
Drawings
Figure 1 is a graph of the solubility of azithromycin in soybean oil after esterification with different fatty acids.
Figure 2 shows the results of the drug time curves of azithromycin tablets, azithromycin fat emulsion and azithromycin long-circulating fat emulsion.
Figure 3 shows the results of the medication time curves of azithromycin injection, azithromycin fat emulsion and azithromycin long-circulating fat emulsion.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings. For the convenience of research and comparison, the following examples, which illustrate azithromycin as a drug substance of a macrolide antibiotic long-circulating emulsion, are described after esterification, but not limited to this. Those skilled in the art will recognize that similar experimental results to those obtained with azithromycin will be obtained using other macrolide antibiotics as the drug substance.
Example 1
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing the formula amount of refined egg yolk lecithin, azithromycin palmitate and PEG synthesized phospholipid, adding a proper amount of ethanol, stirring at 60 ℃ to dissolve the emulsifier, adding the formula amount of refined soybean oil after complete dissolution, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing formula amounts of octadecylamine, glycerin and polysorbate 80, adding a proper amount of water for injection, heating and stirring at 60 ℃ to dissolve or disperse uniformly, and using the mixture as a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 5000rpm/min for 10min at 60 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure for homogenizing particle size, homogenizing at 60 deg.C and 400bar for 2 times, and circulating at 60 deg.C and 800bar for 4 times to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 1.
Table 1 example 1 sample results test table
Example 2
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing the formula amount of refined soybean lecithin, azithromycin laurate and PEG synthesized phospholipid, adding a proper amount of ethanol, stirring at 70 ℃ to dissolve an emulsifier, adding the formula amount of refined soybean oil and medium-chain oil after complete dissolution, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing DOTAP, glycerol and poloxamer 188 according to the prescription amount, adding a proper amount of water for injection, heating and stirring at 70 ℃ to dissolve or disperse uniformly, and using the mixture as a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 7000rpm/min for 15min at 70 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 6 times at 70 deg.C and 800 bar.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 2.
Table 2 example 2 sample results test table
Example 3
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing yolk phosphatidylcholine, azithromycin caprylate and PEG synthesized phospholipid according to the prescription amount, adding a proper amount of ethanol, stirring at 75 ℃ to dissolve an emulsifier, adding refined soybean oil and medium-chain oil after complete dissolution, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing formula amounts of octadecylamine, glycerin and polysorbate 80, adding a proper amount of water for injection, heating and stirring at 75 ℃ to dissolve or disperse uniformly to obtain a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 8000rpm/min for 20min at 75 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 6 times at 70 deg.C and 1000bar homogenizing pressure to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 3.
Table 3 example 3 sample results test table
Example 4
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing 15-hydroxystearic acid polyethylene glycol ester, azithromycin myristate and PEG in the formula amount, adding an appropriate amount of ethanol, stirring at 80 ℃ to dissolve an emulsifier, adding refined soybean oil and olive oil in the formula amount after completely dissolving, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing octadecylamine, glycerol and sodium dodecyl sulfate according to the prescription amount, adding a proper amount of water for injection, heating and stirring at 80 ℃ to dissolve or uniformly disperse the octadecylamine, the glycerol and the sodium dodecyl sulfate to be used as a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 10000rpm/min for 5min at 80 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 4 times at 70 deg.C and 800bar for homogenization to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 4.
Table 4 example 4 sample results test table
Example 5
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing the refined egg yolk lecithin, the azithromycin stearate and the PEG synthetic phospholipid according to the prescription amount, adding a proper amount of ethanol, stirring at 75 ℃ to dissolve the emulsifier, and removing the ethanol through rotary evaporation after complete dissolution to obtain the oil phase.
(2) Weighing the prescription dose of cetyl ammonium bromide, poloxamer 188 and glycerol, adding a proper amount of water for injection, heating and stirring at 75 ℃ to dissolve or disperse uniformly, and using the mixture as a water phase for standby.
(3) Adding the oil phase into the water phase, and shearing at 8000rpm/min for 20min at 75 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 6 times at 70 deg.C and 1000bar homogenizing pressure to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 5.
Table 5 example 5 sample results test table
Example 6
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing the formula amount of refined soybean lecithin, azithromycin laurate and PEG synthesized phospholipid, adding a proper amount of ethanol, stirring at 50 ℃ to dissolve the emulsifier, adding the formula amount of refined soybean oil and medium-chain oil after complete dissolution, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing DOTAP, glycerol and poloxamer 188 according to the prescription amount, adding a proper amount of water for injection, heating and stirring at 50 ℃ to dissolve or disperse uniformly, and using the mixture as a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 3000rpm/min for 5min at 50 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 4 times at 50 deg.C and 200bar homogenizing pressure to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indexes are shown in table 6.
Table 6 example 6 sample results test table
Example 7
The macrolide antibiotic long-circulating emulsion of the present example contains per 1000mL of the emulsion:
the preparation method of the macrolide antibiotic long-circulating emulsion comprises the following steps:
(1) weighing the formula amount of refined soybean lecithin, azithromycin laurate and PEG synthesized phospholipid, adding a proper amount of ethanol, stirring at 80 ℃ to dissolve the emulsifier, adding the formula amount of refined soybean oil and medium-chain oil after complete dissolution, and removing the ethanol through rotary evaporation to obtain an oil phase.
(2) Weighing DOTAP, glycerol and poloxamer 188 according to the prescription amount, adding a proper amount of water for injection, heating and stirring at 80 ℃ to dissolve or disperse uniformly, and using the mixture as a water phase for later use.
(3) Adding the oil phase into the water phase, and shearing at 20000rpm/min for 25min at 80 deg.C to obtain primary emulsion.
(4) Homogenizing the obtained colostrum under high pressure, homogenizing the particle size, and circulating for 10 times at 70 deg.C and 1500bar homogenizing pressure to obtain final emulsion.
(5) Homogenizing under high pressure to obtain final milk, filtering to remove bacteria, bottling, introducing nitrogen gas for protection, and sealing to obtain the final product.
The emulsion of the high proportion of emulsifier prepared in this example was milky white in appearance, good in fluidity, slightly shaken and had a blue opalescence on the wall after high pressure homogenization. The corresponding physicochemical indices are shown in table 7.
Table 7 table for examining results of samples of example 7
Example 8
In order to further investigate the influence of esterification of different fatty acids and azithromycin on the improvement of the solubility of azithromycin in soybean oil, saturated fatty acids such as n-hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid and the like are respectively adopted to be esterified with azithromycin to obtain esterified substances of different azithromycin fatty acids, the saturated solubility of the azithromycin fatty acid esters in soybean oil is measured, and the result is shown in figure 1.
The solubility measurements show that as the carbon chain length of the fatty acid increases, the higher the solubility of the fatty acid ester formed with azithromycin in the oil phase. The reason is that the polarity of the fatty acid is gradually reduced along with the increase of the carbon chain length of the fatty acid, and the polarity of the azithromycin can be reduced to a higher degree after the fatty acid is esterified with the azithromycin, so that the solubility of the fatty acid in an oil phase is increased. The ability of the fatty acids to reduce the polarity of azithromycin is as follows from big to small: stearic acid > palmitic acid > myristic acid > lauric acid > capric acid > pelargonic acid > caprylic acid > heptanoic acid > n-hexanoic acid.
Example 9
The bioavailability of the commercial azithromycin tablets and the bioavailability of the self-made azithromycin long-circulating fat emulsion are compared by adopting a rat intragastric administration method and a tail vein injection method. Taking 24 male SD rats, randomly dividing into 4 groups, 6 rats in each group, respectively orally administering 1 group and intravenously administering 3 groups, and according to an equivalent dose conversion algorithm of body surface area, the equivalent administration dose of the SD rats relative to human is about 44.98mg/kg, the body weight of the rats is calculated according to 200g, and then the administration dose of the rats is 8.996 mg. The commercial azithromycin tablets are ground into powder, 5 percent CMC-Na is used for preparing suspension with the concentration of 4.498mg/mL, and the intragastric volume is 2 mL.
3 groups of intravenous injection administration are respectively an azithromycin injection group, an azithromycin fat emulsion group and an azithromycin long-circulating fat emulsion group, wherein the azithromycin injection is formed by referring to a prescription of the marketed azithromycin injection, and the injection with the azithromycin content of 4.498mg/mL is finally prepared, wherein the ethanol content is 2% (the injection cannot be dissolved when no ethanol is added). The azithromycin fat emulsion is phospholipid synthesized without adding PEG, and the charge regulator is sodium oleate. The content of the medicine is the same as that of the azithromycin intestinal circulating milk. Azithromycin fatty milk and azithromycin intestinal circulating milk according to the drug content of example 1, the administration volume is 0.9mL, 0.5mL of blood is taken from the orbit after administration for 0.083, 0.167, 0.25, 0.333, 0.5, 1, 1.5, 2, 5, 10, 18, 24, 36 and 48 hours, the blood is centrifuged for 10min at 4000rpm, 180 mul of supernatant blood plasma is taken, the blood concentration analysis is carried out after the blood plasma is processed, and the pharmaceutical time curve is drawn, which is shown in figure 2.
As is clear from Table 1, FIG. 2 and FIG. 3, the peak concentration C of the oral administration group after administrationmax23.68mg/mL, which is significantly lower than the group administered by injection. The oral administration group has fast elimination in vivo and half-life period t of the medicine1/23.774 h. Compared with azithromycin injection, the injection has the advantages that the peak reaching time is faster to 0.083h due to the absence of an absorption phase, but the half life period is not greatly different from that of an oral administration group. However, compared with the azithromycin emulsion dosage group, the azithromycin injection has shorter MRT (total body temperature) due to faster metabolism in vivo. And the azithromycin palmitate fat emulsion and the azithromycin palmitate long-circulating injection emulsion are slowly eliminated in vivo, and the half-life periods are respectively 9.47h and 19.14 h. The area under the curve is obviously larger than that of the oral administration group when the medicine is injected, namely, the bioavailability is obviously higher than that of the oral administration group. As can be seen from Table 1, the lower points of the drug administration curves of the azithromycin palmitate fat emulsion and the azithromycin palmitate long-circulating injection emulsion are respectively 2.59 times and 3.76 times of those of the oral administration group, namely, the drug administration by the injection of the emulsion can obviously improve the level of blood concentration.
Meanwhile, compared with the results of an injection administration group, the azithromycin palmitate fat emulsion is added with PEG to synthesize phospholipid, and is changed with a positive charge regulator, so that the in vivo action time of the azithromycin palmitate fat emulsion can be obviously prolonged. The PEG can avoid phagocytosis of a reticuloendothelial cell system in vivo, and the positive charge on the surface of emulsion particles is beneficial to the aggregation of the medicament around bacteria, so that the in vivo circulation time is prolonged, and the medicament concentration of the infected part of the bacteria is increased. Therefore, the azithromycin is prepared into the growth cycle emulsion, so that the dosage can be reduced, the acting time of the medicament in the body can be prolonged, and in addition, the dosage is reduced, the absorption of the medicament is improved, and the reduction of the in-vivo drug resistance of the macrolide antibiotics is facilitated.
TABLE 1 comparison of the pharmacokinetic parameters of different azithromycin dosage forms
The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, therefore, the present invention is not limited by the appended claims.
Claims (10)
1. The macrolide antibiotic long-circulating emulsion is characterized by comprising the following components in percentage by mass: 0.1-1% of macrolide antibiotics, 10-30% of oil phase solvent, 0.5-5% of emulsifier, 0.2-5% of co-emulsifier, 0.3-5% of potential regulator, 1-5% of osmotic pressure regulator and the balance of water for injection; the structure of the macrolide antibiotic comprises-NH2One or more of a group, -OH group; before use, the macrolide antibiotics are esterified to form macrolide antibiotic fatty acid esters.
2. The macrolide antibiotic long-circulating emulsion of claim 1, wherein the emulsifier comprises a pegylated synthetic phospholipid.
3. The macrolide antibiotic long-circulating emulsion according to claim 1, wherein said esterification treatment is performed by: and (3) carrying out esterification reaction with the macrolide antibiotic by using saturated fatty acid to form the macrolide antibiotic fatty acid ester.
4. The macrolide antibiotic long-circulating emulsion according to claim 3, wherein the carbon chain length of the saturated fatty acid is 6 to 18.
5. The macrolide antibiotic long-circulating emulsion of claim 4, wherein the saturated fatty acid comprises one of caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid.
6. The macrolide antibiotic long-circulating emulsion of claim 1, wherein the potential regulator is a positive charge regulator.
7. The macrolide antibiotic long circulation emulsion according to claim 1, wherein the macrolide antibiotic is one of azithromycin, clarithromycin, roxithromycin, dirithromycin, flurithromycin, and rotamycin; the oil phase solvent comprises one or more of fatty acid triglyceride; the emulsifier also comprises one or more of egg yolk phospholipid, soybean phospholipid, phosphatidylcholine and 15-hydroxystearic acid polyethylene glycol ester; the auxiliary emulsifier comprises one or more of polysorbate 80, poloxamer 188, sodium dodecyl sulfate and hexadecyl ammonium bromide; the potential regulator comprises one or more of octadecylamine, DOTAP and cationic surfactant; the osmotic pressure regulator comprises one or more of glycerol and mannitol.
8. The nutritional emulsion of claim 7 wherein the oil phase solvent comprises one or more of soybean oil, olive oil, tea seed oil, fish oil, castor oil, coix seed oil, tricaprylin, tricaprin, caprylocapran, trilaurin.
9. A process for the preparation of a macrolide antibiotic long-circulating emulsion according to claim 1, comprising the steps of:
A. adding the macrolide antibiotics and the emulsifier into the oil phase solvent according to the proportion, continuously heating and stirring until the macrolide antibiotics and the emulsifier are completely dissolved to form a drug-loaded oil phase;
B. mixing the potential regulator, the osmotic pressure regulator and the co-emulsifier with the water according to the prescription composition, and heating until the mixture is completely dissolved to form a water phase;
C. slowly adding the drug-loaded oil phase into the water phase, shearing at a high speed under stirring, and adding the rest water to prepare primary emulsion;
D. homogenizing the primary emulsion under high pressure to obtain final emulsion;
E. and (3) sterilizing the final emulsion, filling nitrogen and sealing to obtain the macrolide antibiotic long-circulating emulsion.
10. The method for preparing a macrolide antibiotic long-circulating emulsion according to claim 9, wherein in the step a, the heating temperature is 50 to 80 ℃; in the step B, the heating temperature is 50-80 ℃; in the step C, the stirring speed of high-speed shearing is 3000-20000 rpm, the shearing time is 5-25 min, and the shearing temperature is 50-70 ℃; in the step D, the high-pressure homogenization specifically comprises the following steps: controlling the homogenizing temperature to be 50-80 ℃, and circulating for 4-10 times under the pressure of 200-1500 bar; in the step E, the sterilization comprises one of filtration sterilization or high-temperature sterilization.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1660872A (en) * | 2005-01-02 | 2005-08-31 | 李�杰 | Antibiotic 2-hydroxy ester in new type macrolide group, preparation and preparing method |
| CN101032467A (en) * | 2007-04-13 | 2007-09-12 | 西安力邦制药有限公司 | Superregulated long-cycled lipid emulsion carrying medicine reagent for mainline |
| CN102784104A (en) * | 2012-07-24 | 2012-11-21 | 宋金春 | Erythromycin ethylsuccinate emulsion injection and its preparation method |
| US20120309780A1 (en) * | 2005-04-12 | 2012-12-06 | Wisconsin Alumni Research Foundation | Micelle composition of polymer and passenger drug |
| CN105188670A (en) * | 2013-03-15 | 2015-12-23 | 微分药物发展联合有限公司 | Emulsion formulations |
| CN108310390A (en) * | 2018-04-10 | 2018-07-24 | 沈阳药科大学 | Docetaxel linoleate and its fat emulsion and application |
| TW201934116A (en) * | 2018-02-14 | 2019-09-01 | 大葉大學 | Nanoemulsion composition including citral and manufacturing method thereof |
-
2020
- 2020-03-13 CN CN202010176898.3A patent/CN111358754A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1660872A (en) * | 2005-01-02 | 2005-08-31 | 李�杰 | Antibiotic 2-hydroxy ester in new type macrolide group, preparation and preparing method |
| US20120309780A1 (en) * | 2005-04-12 | 2012-12-06 | Wisconsin Alumni Research Foundation | Micelle composition of polymer and passenger drug |
| CN101032467A (en) * | 2007-04-13 | 2007-09-12 | 西安力邦制药有限公司 | Superregulated long-cycled lipid emulsion carrying medicine reagent for mainline |
| CN102784104A (en) * | 2012-07-24 | 2012-11-21 | 宋金春 | Erythromycin ethylsuccinate emulsion injection and its preparation method |
| CN105188670A (en) * | 2013-03-15 | 2015-12-23 | 微分药物发展联合有限公司 | Emulsion formulations |
| TW201934116A (en) * | 2018-02-14 | 2019-09-01 | 大葉大學 | Nanoemulsion composition including citral and manufacturing method thereof |
| CN108310390A (en) * | 2018-04-10 | 2018-07-24 | 沈阳药科大学 | Docetaxel linoleate and its fat emulsion and application |
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