A pharmaceutical composition formulation containing ertapenem and its preparation method thereof

Abstract

This invention belongs to the field of biomedical technology, specifically relating to a pharmaceutical composition formulation containing ertapenem and its preparation method. The ertapenem-containing pharmaceutical composition formulation comprises the following raw materials: 10-20 parts ertapenem, 40-60 parts choline-leucine ionic liquid, 30-50 parts polyethylene glycol-polylactic acid-glycolic acid copolymer, 2-5 parts gluconolactone, 2-3 parts antioxidant, 40-50 parts deionized water, 3-5 parts surfactant, and 20-30 parts mannitol. This invention utilizes choline-leucine ionic liquid to enhance the solubility and stability of the polyethylene glycol-polylactic acid-glycolic acid copolymer with ertapenem, forming uniform micelles to protect the drug from hydrolysis. Liquid nitrogen freezing technology is used to improve preparation efficiency and product quality, ensuring biocompatibility.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a pharmaceutical composition preparation containing ertapenem and a preparation method thereof. Background Art

[0002] Ertapenem, as a highly effective carbapenem antibiotic, occupies an important position in clinical treatment due to its broad-spectrum antibacterial activity. It not only has strong antibacterial efficacy against a variety of Gram-positive and Gram-negative bacteria, but is also particularly suitable for combating drug-resistant strains, such as methicillin-resistant Staphylococcus aureus (MRSA) and drug-resistant Gram-negative bacilli. The mechanism of action of ertapenem is to inhibit penicillin-binding proteins (PBPs), a key enzyme in bacterial cell wall synthesis, thereby destroying the cell wall structure and causing bacterial death. Due to its broad antibacterial spectrum and effectiveness against drug-resistant strains, ertapenem has important application value in the treatment of severe infections.

[0003] At present, Ertapenem is mainly provided in the form of powder for injection, which needs to be administered by intravenous or intramuscular injection. Although this mode of administration can ensure that the drug quickly enters the blood circulation and reaches the blood drug concentration required for treatment, it also has some limitations and shortcomings: the powder dosage form for injection is easily affected by environmental factors such as temperature and humidity during storage and transportation and degraded. For example, in a high temperature or high humidity environment, the drug may undergo hydrolysis, resulting in a decrease in the content of active ingredients, thereby affecting the stability of the drug; in addition to degradation, the powder dosage form for injection may also undergo physicochemical changes, such as crystal transformation, aggregation or agglomeration, etc. These changes will also affect the solubility and bioavailability of the drug. Therefore, based on the above problems, it is extremely necessary to develop an Ertapenem pharmaceutical composition preparation with excellent biocompatibility and good stability. Summary of the invention

[0004] In view of the defects of the prior art, the object of the present invention is to provide a pharmaceutical composition preparation containing ertapenem and a preparation method thereof.

[0005] The technical effect described in the present invention is achieved through the following technical scheme: a pharmaceutical composition preparation containing ertapenem, which comprises the following raw materials in parts by weight: 10 to 20 parts of ertapenem, 40 to 60 parts of choline-leucine ionic liquid, 30 to 50 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 2 to 5 parts of gluconolactone, 2 to 3 parts of antioxidant, 40 to 50 parts of deionized water, 3 to 5 parts of surfactant and 20 to 30 parts of mannitol.

[0006] Preferably, the specific preparation steps of the choline-leucine ionic liquid are as follows:

[0007] A1: Mix choline chloride and leucine in equal molar masses in deionized water, stir at 300-500 rpm with a magnetic stirrer at room temperature for 60-90 min, cool naturally to room temperature, centrifuge, and filter to obtain a mixture;

[0008] A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 10-20 mbar, the temperature to 40-60°C, the rotation speed to 200-300 rpm, the condenser temperature to 0-5°C, evaporate for 1-2 hours, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product;

[0009] A3: The purified product prepared in step A2 is added to anhydrous ethanol and dispersed evenly by 30W ultrasonic treatment to obtain choline-leucine ionic liquid.

[0010] Preferably, in step A1, the ratio of the total amount of choline chloride and leucine to the amount of deionized water is 0.01 mol: 30-50 mL.

[0011] Preferably, the ratio of the purified product to anhydrous ethanol in step A3 is 1 g:10-15 mL.

[0012] Preferably, the specific preparation steps of the polyethylene glycol-polylactic acid-glycolic acid copolymer are as follows:

[0013] (1): Dissolve polyethylene glycol in dichloromethane, stir evenly, slowly add an equal amount of ethanol, continue stirring to dissolve evenly, slowly add concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature at 60-70°C, continue stirring to react for 4-6 hours, and obtain a reaction mixture; then add the reaction mixture to 1 volume of 0°C cold ethanol, filter, wash until neutral, and vacuum dry at 60°C to constant weight to obtain activated polyethylene glycol;

[0014] (2): Add polylactic acid-glycolic acid copolymer to dichloromethane, stir and mix evenly, add the activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature at 60-80°C, react for 12-24 hours, then add to 1 volume of 0°C cold ethanol, filter, wash to neutrality, and vacuum dry at 60°C to constant weight to obtain polyethylene glycol-polylactic acid-glycolic acid copolymer.

[0015] Preferably, the ratio of the amount of polyethylene glycol, dichloromethane and concentrated sulfuric acid used in step (1) is 1 g:10-20 mL:0.05-0.1 mL.

[0016] Preferably, in step (2), the ratio of the amount of the polylactic acid-glycolic acid copolymer, dichloromethane and activated polyethylene glycol is 1 g: 10-15 mL: 0.2-0.5 g.

[0017] Preferably, the antioxidant is any one of vitamin C, vitamin E, butylated hydroxyanisole and glutathione.

[0018] Preferably, the surfactant is any one of Tween-80, polyvinyl pyrrolidone, sodium lauryl sulfate and cetyltrimethylammonium bromide.

[0019] Preferably, another aspect of the present invention is to provide a method for preparing a pharmaceutical composition preparation containing ertapenem, and the specific preparation steps are as follows:

[0020] Step 1: Add polyethylene glycol-polylactic acid-glycolic acid copolymer to choline-leucine ionic liquid, stir and dissolve evenly, then add ertapenem, continue stirring until evenly dispersed, control the temperature at 25-30° C., perform ultrasonic treatment, set the parameters to 30-35 kHz, power to 20-30 W, pause for 1 minute after each treatment, and the treatment time is 5-15 minutes in total to obtain a mixed solution;

[0021] Step 2: adding gluconolactone, antioxidant, surfactant and mannitol to the mixed solution prepared in step 1, stirring and mixing evenly, then gradually and slowly adding deionized water dropwise during ultrasonic treatment, and ultrasonically treating at a power of 30 to 50 W for 15 to 30 minutes to obtain a micellar solution;

[0022] Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

[0023] The beneficial effects of the present invention are as follows:

[0024] The present invention utilizes the good solubility of choline-leucine ionic liquid to effectively dissolve polyethylene glycol-polylactic acid-glycolic acid copolymer (PEG-PLGA) and ertapenem. The polarity and hydrogen bond donor / acceptor properties of the ionic liquid enhance the solubility of ertapenem in the solution, and its low volatility and high thermal stability provide a stable environment during the preparation process, reduce the influence of temperature and solvent volatilization on the system, and thus improve the stability of the entire preparation. The choline-leucine ionic liquid provides a uniform solution environment, ensures the uniform distribution of ertapenem in the polyethylene glycol-polylactic acid-glycolic acid copolymer, and forms micelles with uniform structure; considering that the hydrophilicity of ertapenem is easily affected by hydrolysis, the polyethylene glycol-polylactic acid-glycolic acid copolymer is an amphiphilic molecule, and forms a micelle structure through self-assembly, the hydrophilic polyethylene glycol part faces the outside, forming a stable aqueous phase environment, and the hydrophobic polylactic acid-glycolic acid copolymer part faces the inside, which can effectively wrap and protect ertapenem, prevent its degradation in the external environment, and further improve the stability of the drug. Choline and leucine in the ionic liquid promote the self-assembly process of polyethylene glycol-polylactic acid-co-glycolic acid through hydrogen bonding and electrostatic interactions, forming a more stable and uniform micelle structure. The hydrophilic polyethylene glycol (PEG) part faces inward, forming a hydrophilic core environment that can effectively encapsulate and protect the hydrophilic ertapenem, while the hydrophobic polylactic acid-co-glycolic acid (PLGA) part faces outward, forming a stable shell to prevent the degradation of ertapenem in the external environment. Leucine, as a hydrophobic amino acid, interacts with the polylactic acid-co-glycolic acid part to further enhance the stability of the micelle structure.

[0025] During the preparation process, choline-leucine ionic liquid is used as an intermediate solvent to improve the efficiency of self-assembly and the consistency of the product. Then deionized water is used as an anti-solvent, combined with ultrasonic oscillation technology to ensure that no dissolved solids or ions remain in the process of preparing micelles, while reducing surface tension, promoting the formation of fine micelles, and accelerating the mixing of drugs and polymers. After removing the ionic liquid, the micelles formed by polyethylene glycol-polylactic acid-glycolic acid copolymer can still maintain their structure, control the release rate of ertapenem, and prolong the action time of the drug. The final product has good biocompatibility, the polyethylene glycol part has anti-immunogenicity, and the polylactic acid-glycolic acid copolymer is a biodegradable material, which ensures the biocompatibility of the entire preparation. In addition, the use of liquid nitrogen freezing hydrosol technology can effectively reduce the overall process time of freeze-drying, avoid the large ice crystal phenomenon that is prone to occur in conventional freezing processes, and prepare a more uniform frozen hydrosol, which prevents ertapenem from decomposing or denaturing during the drying process, thereby improving the yield of the product.

[0026] In summary, the present invention not only improves the solubility and stability of the ertapenem preparation through the synergistic effect of multiple innovative technologies, but also improves the preparation efficiency and product quality and ensures biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 It is a biocompatibility test diagram of the pharmaceutical composition preparation of ertapenem prepared in Examples 1 to 4 of the present invention;

[0029] Figure 2 It is a graph showing the in vitro dissolution test of the pharmaceutical composition preparations of ertapenem prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2;

[0030] Figure 3 It is a stability test chart of the pharmaceutical composition preparation of ertapenem prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2;

[0031] Figure 4 is a SEM scanning electron microscope image of the pharmaceutical composition preparation of ertapenem prepared in Example 2 of the present invention;

[0032] Figure 5 It is the FTIR infrared spectrum of the pharmaceutical composition preparation of ertapenem prepared in Example 2 of the present invention. DETAILED DESCRIPTION

[0033] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. It should be noted that, unless otherwise specified, the raw materials involved in the present invention are purchased through conventional commercial channels.

[0034] Example 1: A pharmaceutical composition preparation containing ertapenem, comprising the following raw materials in parts by weight: 10 parts of ertapenem, 40 parts of choline-leucine ionic liquid, 30 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 2 parts of gluconolactone, 2 parts of antioxidant, 40 parts of deionized water, 3 parts of surfactant and 20 parts of mannitol.

[0035] 1. Preparation of choline-leucine ionic liquid:

[0036] A1: 0.05 mol of choline chloride and 0.05 mol of leucine were mixed in 300 mL of deionized water, and stirred at 300 rpm with a magnetic stirrer at room temperature for 90 min. After stirring, the mixture was naturally cooled to room temperature, centrifuged, and filtered to obtain a mixture;

[0037] A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 10 mbar, the temperature to 60°C, the speed to 200 rpm, the condenser temperature to 5°C, evaporate for 2 h, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product;

[0038] A3: Add 20 g of the purified product prepared in step A2 to 200 mL of anhydrous ethanol, and disperse evenly by 30 W ultrasonic treatment to obtain choline-leucine ionic liquid;

[0039] 2. Preparation of Polyethylene Glycol-Polylactic Acid-Co-glycolic Acid Copolymer:

[0040] (1): Dissolve 10 g of polyethylene glycol in 100 mL of dichloromethane, stir evenly, slowly add 100 mL of ethanol, continue stirring to dissolve evenly, slowly add 0.5 mL of concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature to 60°C, continue stirring to react for 6 hours, and obtain a reaction mixture; then add the reaction mixture into 200 mL of 0°C cold ethanol, filter, wash until neutral, and vacuum dry at 60°C to constant weight to obtain activated polyethylene glycol;

[0041] (2): Add 20 g of polylactic acid-glycolic acid copolymer to 200 mL of dichloromethane, stir and mix well, add 4 g of activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature to 60° C., react for 24 h, then add 200 mL of 0° C. cold ethanol, filter, wash to neutrality, and vacuum dry at 60° C. to constant weight to obtain polyethylene glycol-polylactic acid-glycolic acid copolymer;

[0042] 3. Preparation of a pharmaceutical composition containing ertapenem:

[0043] Step 1: Add 30 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer to 40 parts of choline-leucine ionic liquid, stir and dissolve evenly, then add 10 parts of ertapenem, continue stirring until evenly dispersed, control the temperature at 25°C, perform ultrasonic treatment, set the parameters to 30kHz, power to 20W, pause for 1 minute after each treatment, and the treatment time is 15 minutes in total to obtain a mixed solution;

[0044] Step 2: Add 2 parts of gluconolactone, 2 parts of vitamin C, 3 parts of polyvinyl pyrrolidone and 20 parts of mannitol to the mixed solution prepared in step 1, stir and mix evenly, then gradually and slowly add 40 parts of deionized water during ultrasonic treatment, and ultrasonically treat at 30 W power for 30 minutes to obtain a micellar solution;

[0045] Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

[0046] Example 2: A pharmaceutical composition preparation containing ertapenem, comprising the following raw materials in parts by weight: 18 parts of ertapenem, 55 parts of choline-leucine ionic liquid, 45 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 4 parts of gluconolactone, 3 parts of antioxidant, 45 parts of deionized water, 4 parts of surfactant and 28 parts of mannitol.

[0047] 1. Preparation of choline-leucine ionic liquid:

[0048] A1: 0.05 mol of choline chloride and 0.05 mol of leucine were mixed in 450 mL of deionized water, and stirred at 450 rpm with a magnetic stirrer for 70 min at room temperature. After stirring, the mixture was naturally cooled to room temperature, centrifuged, and filtered to obtain a mixture;

[0049] A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 20 mbar, the temperature to 40°C, the speed to 300 rpm, the condenser temperature to 0°C, evaporate for 2 h, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product;

[0050] A3: Add 20 g of the purified product prepared in step A2 to 280 mL of anhydrous ethanol, and disperse evenly by 30 W ultrasonic treatment to obtain choline-leucine ionic liquid;

[0051] 2. Preparation of Polyethylene Glycol-Polylactic Acid-Co-glycolic Acid Copolymer:

[0052] (1): Dissolve 10 g of polyethylene glycol in 180 mL of dichloromethane, stir evenly, slowly add 180 mL of ethanol, continue stirring to dissolve evenly, slowly add 0.8 mL of concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature to 70°C, continue stirring to react for 5 hours, and obtain a reaction mixture; then add the reaction mixture into 360 mL of 0°C cold ethanol, filter, wash until neutral, and vacuum dry at 60°C to constant weight to obtain activated polyethylene glycol;

[0053] (2): Add 20 g of polylactic acid-glycolic acid copolymer to 280 mL of dichloromethane, stir and mix well, then add 9 g of activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature to 75° C., react for 20 h, then add 280 mL of 0° C. cold ethanol, filter, wash to neutrality, and vacuum dry at 60° C. to constant weight to obtain polyethylene glycol-polylactic acid-glycolic acid copolymer;

[0054] 3. Preparation of a pharmaceutical composition containing ertapenem:

[0055] Step 1: Add 45 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer to 55 parts of choline-leucine ionic liquid, stir and dissolve evenly, then add 18 parts of ertapenem, continue stirring until evenly dispersed, control the temperature at 27°C, perform ultrasonic treatment, set the parameters to 35kHz, power to 25W, pause for 1min after each treatment, and the treatment time is 10min in total to obtain a mixed solution;

[0056] Step 2: Add 4 parts of gluconolactone, 3 parts of vitamin E, 4 parts of Tween-80 and 28 parts of mannitol to the mixed solution prepared in step 1, stir and mix evenly, then gradually and slowly add 45 parts of deionized water during ultrasonic treatment, and ultrasonically treat at a power of 40 W for 25 minutes to obtain a micellar solution;

[0057] Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

[0058] Example 3: A pharmaceutical composition preparation containing ertapenem, comprising the following raw materials in parts by weight: 15 parts of ertapenem, 50 parts of choline-leucine ionic liquid, 40 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 3 parts of gluconolactone, 2 parts of antioxidant, 45 parts of deionized water, 4 parts of surfactant and 25 parts of mannitol.

[0059] 1. Preparation of choline-leucine ionic liquid:

[0060] A1: 0.05 mol of choline chloride and 0.05 mol of leucine were mixed in 400 mL of deionized water, and stirred at 400 rpm with a magnetic stirrer at room temperature for 80 min. After stirring, the mixture was naturally cooled to room temperature, centrifuged, and filtered to obtain a mixture;

[0061] A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 15 mbar, the temperature to 50°C, the speed to 240 rpm, the condenser temperature to 3°C, evaporate for 1.5 h, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product;

[0062] A3: Add 20 g of the purified product prepared in step A2 to 250 mL of anhydrous ethanol, and disperse evenly by 30 W ultrasonic treatment to obtain choline-leucine ionic liquid;

[0063] 2. Preparation of Polyethylene Glycol-Polylactic Acid-Co-glycolic Acid Copolymer:

[0064] (1): Dissolve 10 g of polyethylene glycol in 150 mL of dichloromethane, stir evenly, slowly add 150 mL of ethanol, continue stirring to dissolve evenly, slowly add 0.7 mL of concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature to 65°C, continue stirring to react for 5 hours, and obtain a reaction mixture; then add the reaction mixture into 300 mL of 0°C cold ethanol, filter, wash until neutral, and vacuum dry at 60°C to constant weight to obtain activated polyethylene glycol;

[0065] (2): Add 20 g of polylactic acid-co-glycolic acid to 250 mL of dichloromethane, stir and mix well, add 7 g of activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature to 70° C., react for 18 h, then add 250 mL of 0° C. cold ethanol, filter, wash to neutrality, and vacuum dry at 60° C. to constant weight to obtain polyethylene glycol-polylactic acid-co-glycolic acid;

[0066] 3. Preparation of a pharmaceutical composition containing ertapenem:

[0067] Step 1: Add 40 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer to 50 parts of choline-leucine ionic liquid, stir and dissolve evenly, then add 15 parts of ertapenem, continue stirring until evenly dispersed, control the temperature at 27°C, perform ultrasonic treatment, set the parameters to 32kHz, power to 25W, pause for 1 minute after each treatment, and the treatment time is 10 minutes in total to obtain a mixed solution;

[0068] Step 2: Add 3 parts of gluconolactone, 2 parts of butylated hydroxyanisole, 4 parts of sodium dodecyl sulfate and 25 parts of mannitol to the mixed solution prepared in step 1, stir and mix evenly, then gradually and slowly add 45 parts of deionized water during ultrasonic treatment, and ultrasonically treat at a power of 40 W for 20 minutes to obtain a micellar solution;

[0069] Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

[0070] Example 4: A pharmaceutical composition preparation containing ertapenem, comprising the following raw materials in parts by weight: 20 parts of ertapenem, 60 parts of choline-leucine ionic liquid, 50 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 5 parts of gluconolactone, 3 parts of antioxidant, 50 parts of deionized water, 5 parts of surfactant and 30 parts of mannitol.

[0071] 1. Preparation of choline-leucine ionic liquid:

[0072] A1: 0.05 mol of choline chloride and 0.05 mol of leucine were mixed in 500 mL of deionized water, and stirred at 500 rpm with a magnetic stirrer at room temperature for 60 min. After stirring, the mixture was naturally cooled to room temperature, centrifuged, and filtered to obtain a mixture;

[0073] A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 20 mbar, the temperature to 60°C, the speed to 200 rpm, the condenser temperature to 0°C, evaporate for 1 h, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product;

[0074] A3: Add 20 g of the purified product prepared in step A2 to 300 mL of anhydrous ethanol, and disperse evenly by 30 W ultrasonic treatment to obtain choline-leucine ionic liquid;

[0075] 2. Preparation of Polyethylene Glycol-Polylactic Acid-Co-glycolic Acid Copolymer:

[0076] (1): Dissolve 10 g of polyethylene glycol in 200 mL of dichloromethane, stir evenly, slowly add 200 mL of ethanol, continue to stir and dissolve evenly, slowly add 1 mL of concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature to 70°C, continue to stir and react for 4 hours to obtain a reaction mixture; then add the reaction mixture into 400 mL of 0°C cold ethanol, filter, wash until neutral, and vacuum dry at 60°C to constant weight to obtain activated polyethylene glycol;

[0077] (2): Add 20 g of polylactic acid-glycolic acid copolymer to 300 mL of dichloromethane, stir and mix well, add 10 g of activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature to 80° C., react for 12 h, then add 300 mL of 0° C. cold ethanol, filter, wash to neutrality, and vacuum dry at 60° C. to constant weight to obtain polyethylene glycol-polylactic acid-glycolic acid copolymer;

[0078] 3. Preparation of a pharmaceutical composition containing ertapenem:

[0079] Step 1: Add 50 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer to 60 parts of choline-leucine ionic liquid, stir and dissolve evenly, then add 20 parts of ertapenem, continue stirring until evenly dispersed, control the temperature at 30°C, perform ultrasonic treatment, set the parameters to 35kHz, power to 30W, pause for 1min after each treatment, and the treatment time is 5min in total to obtain a mixed solution;

[0080] Step 2: Add 5 parts of gluconolactone, 3 parts of glutathione, 5 parts of hexadecyltrimethylammonium bromide and 30 parts of mannitol to the mixed solution prepared in step 1, stir and mix evenly, then gradually and slowly add 50 parts of deionized water during ultrasonic treatment, and ultrasonically treat at a power of 50 W for 15 minutes to obtain a micellar solution;

[0081] Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

[0082] Comparative Example 1: The operation of Comparative Example 1 is substantially the same as that of Example 2, except that anhydrous ethanol is used as a solvent instead of the choline-leucine ionic liquid in Comparative Example 1.

[0083] Comparative Example 2: The operation of Comparative Example 2 is substantially the same as that of Example 3, except that, in Comparative Example 2, polylactic acid-glycolic acid copolymer is used instead of polyethylene glycol-polylactic acid-glycolic acid copolymer.

[0084] Performance Test:

[0085] Biocompatibility: 10 mL of fibroblasts in good logarithmic growth phase were selected, digested with 0.25 wt% trypsin and centrifuged, and then resuspended in 10 mL of DMEM culture medium containing 10% fetal bovine serum; the fibroblasts were inoculated in a 96-well culture plate, with a cell number of about 5 × 10 per well. 4A blank control without cells was set up, and the edge wells were filled with sterile PBS and cultured overnight at 37°C in a CO2 environment with a volume fraction of 5%; samples (the pharmaceutical composition preparation of ertapenem prepared in Examples 1 to 4 of the present invention) were added respectively, and the samples were diluted to 10 mg / mL with PBS, and a gentamicin solution with 100 μg / mL was added as a positive control group, and an untreated group was used as a cell control group. Each group had 6 replicate wells, and 20 μL was added to each well, and then cultured for 48 hours at 37°C and 5% CO2. 4 hours before the end of the culture, an MTT solution diluted to 1 mg / mL with PBS was added, and the culture was terminated after continuing to culture for 4 hours; the liquid in the well was discarded, 100 μL of DMSO was added to dissolve the crystals, and the absorbance of each well was measured at 570 nm by an M3 plate reader. Cell survival rate (%) = (absorbance of the test group - absorbance of the control group) / (absorbance of the treatment group - absorbance of the control group) × 100%. The test results are as follows Figure 1 shown.

[0086] Depend on Figure 1 The results show that the pharmaceutical composition preparation of ertapenem prepared in the present invention has excellent biocompatibility.

[0087] In vitro dissolution release test: 100 mg of the pharmaceutical composition preparation of ertapenem prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2 was dissolved in 90 mL of pH 6.8 phosphate buffer, the temperature was controlled at 37°C, and samples were taken at 0.5 h, 1 h, 2 h, 4 h, 6 h, 12 h, and 24 h, 10 mL of sample was taken at each sampling point, and then 10 mL of solution was replenished to determine the release amount (Xn'=Xn+(X1+X2+X n-1 )V2 / V1; where X n is the relative percentage dissolution measured after correction for the nth time, X n’ is the relative percentage dissolution actually measured for the nth time, V1 is the volume of the dissolution medium, and V2 is the volume added after each sampling). The results are as follows Figure 2 shown.

[0088] Depend on Figure 2 As a result, it can be seen from the results of Comparative Example 1 and Example 2 that the solubility of anhydrous ethanol is not as strong as that of choline-leucine ionic liquid, and anhydrous ethanol has a higher volatility, which affects the stability of the entire preparation. Anhydrous ethanol lacks the promoting effect of choline-leucine ionic liquid, which affects the stability and uniformity of the micelle structure. Although anhydrous ethanol will dissolve faster in the early stage, its solubility performance is not as good as that of choline-leucine ionic liquid; from the results of Comparative Example 2 and Example 3, it can be seen that the lack of hydrophilicity of the polyethylene glycol part leads to an unstable micelle structure, and ertapenem is more likely to contact the dissolution medium, resulting in a large amount of dissolution in the early stage.

[0089] Stability test: The pharmaceutical composition preparation samples of ertapenem prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2 were sealed according to commercial packaging and placed at a temperature of 35 ± 2°C and a relative humidity of 65 ± 5% for 3 months. Samples were taken once in the first, second and third months of the test period to test the solution pH. The average value of the three samples was taken and the appearance changes of the preparations were recorded. The pH change results are shown in FIG. Figure 3 The appearance changes are shown in Table 1 below.

[0090] Table 1. Stability test results of the pharmaceutical composition preparation of ertapenem

[0091]

[0092] From Table 1 and Figure 3 As a result, the pharmaceutical composition preparation of ertapenem prepared by the present invention has excellent stability in the accelerated long-term storage test, and the pH change is stable; from the results of Comparative Example 1 and Example 2, it can be seen that anhydrous ethanol is used as a solvent to replace the choline-leucine ionic liquid, and the dissolution ability of anhydrous ethanol is not as strong as that of the choline-leucine ionic liquid, resulting in reduced solubility of polyethylene glycol-polylactic acid-glycolic acid copolymer and ertapenem, and lacks hydrogen bonds and electrostatic interactions in the choline-leucine ionic liquid, and the stability and uniformity of the micelle structure are significantly affected; from the results of Comparative Example 2 and Example 3, it can be seen that only using polylactic acid-glycolic acid copolymer may not form a stable micelle structure like polyethylene glycol-polylactic acid-glycolic acid copolymer, and lacks the protection provided by polyethylene glycol, and ertapenem is easily affected by hydrolysis, resulting in agglomeration.

[0093] Spectrum test: The spectrum of the pharmaceutical composition sample of ertapenem prepared in Example 2 was tested using a scanning electron microscope. The results are as follows Figure 4 As shown; FTIR spectrometer tested the sample spectrum of the pharmaceutical composition preparation of ertapenem, and the results were as follows Figure 5 shown.

[0094] Depend on Figure 4 The results show that the particles in the pharmaceutical composition preparation of ertapenem prepared by the present invention have uniform morphology and relatively consistent size distribution, showing a typical spherical structure, a relatively smooth particle surface, a clear interface, and an obvious micelle structure; Figure 5 The results show that the FTIR spectrum shows obvious COC stretching vibration peaks of the ether bond of polyethylene glycol, NH stretching vibration peaks of the amine group of ertapenem, and C=O stretching vibration peaks of PEG-PLGA and ertapenem. Based on the above results, the ertapenem prepared by the present invention is effectively encapsulated to form a micellar structure.

[0095] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a pharmaceutical composition preparation containing ertapenem, characterized in that: The pharmaceutical composition preparation comprises the following raw materials in parts by weight: 10 to 20 parts of ertapenem, 40 to 60 parts of choline-leucine ionic liquid, 30 to 50 parts of polyethylene glycol-polylactic acid-glycolic acid copolymer, 2 to 5 parts of gluconolactone, 2 to 3 parts of antioxidant, 40 to 50 parts of deionized water, 3 to 5 parts of surfactant and 20 to 30 parts of mannitol; The specific preparation steps of the choline-leucine ionic liquid are as follows: A1: Mix choline chloride and leucine in equal molar masses in deionized water, stir at room temperature, cool naturally to room temperature, centrifuge, and filter to obtain a mixture; A2: Add the mixture prepared in step A1 into a rotary evaporator, control the pressure to 10-20 mbar, the temperature to 40-60°C, the rotation speed to 200-300 rpm, the condenser temperature to 0-5°C, evaporate for 1-2 hours, wash with cold anhydrous ethanol, and freeze-dry at -60°C in a vacuum until constant weight to obtain a purified product; A3: adding the purified product prepared in step A2 into anhydrous ethanol, and dispersing it evenly by ultrasonic treatment to obtain choline-leucine ionic liquid; The ratio of the total amount of choline chloride and leucine to deionized water in step A1 is 0.01 mol: 30-50 mL; the ratio of the purified product to anhydrous ethanol in step A3 is 1 g: 10-15 mL; The specific preparation steps of the polyethylene glycol-polylactic acid-glycolic acid copolymer are as follows: (1): Dissolve polyethylene glycol in dichloromethane, stir evenly, slowly add an equal amount of ethanol, continue stirring to dissolve evenly, slowly add concentrated sulfuric acid while continuing to stir, heat in a water bath to control the temperature, continue stirring to react, and obtain a reaction mixture; then add the reaction mixture into 0°C cold ethanol, filter, wash to neutrality, and vacuum dry to constant weight to obtain activated polyethylene glycol; (2): Add polylactic acid-glycolic acid copolymer to dichloromethane, stir and mix evenly, then add the activated polyethylene glycol prepared in step (1), heat in a water bath to control the temperature, and after the reaction is completed, add to 0°C cold ethanol, filter, wash to neutrality, and vacuum dry to constant weight to obtain polyethylene glycol-polylactic acid-glycolic acid copolymer; The ratio of the amount of polyethylene glycol, dichloromethane and concentrated sulfuric acid used in step (1) is 1g:10-20mL:0.05-0.1mL; the ratio of the amount of polylactic acid-glycolic acid copolymer, dichloromethane and activated polyethylene glycol used in step (2) is 1g:10-15mL:0.2-0.5g; The antioxidant is any one of vitamin C, vitamin E, butylated hydroxyanisole and glutathione; The surfactant is any one of Tween-80, polyvinyl pyrrolidone, sodium lauryl sulfate and hexadecyltrimethylammonium bromide; The specific preparation steps of the pharmaceutical composition preparation are as follows: Step 1: Add polyethylene glycol-polylactic acid-glycolic acid copolymer to choline-leucine ionic liquid, stir and dissolve evenly, then add ertapenem, continue stirring until evenly dispersed, control the temperature at 25-30° C., perform ultrasonic treatment, set the parameters to 30-35 kHz, power to 20-30 W, pause for 1 minute after each treatment, and the treatment time is 5-15 minutes in total to obtain a mixed solution; Step 2: adding gluconolactone, antioxidant, surfactant and mannitol to the mixed solution prepared in step 1, stirring and mixing evenly, then gradually and slowly adding deionized water dropwise during ultrasonic treatment, and ultrasonically treating at a power of 30 to 50 W for 15 to 30 minutes to obtain a micellar solution; Step 3: Add the micelle solution prepared in step 2 into liquid nitrogen and rapidly cool it to obtain a frozen hydrogel, which is then vacuum dried at -40°C to a constant weight to obtain a pharmaceutical composition preparation.

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