Application of naringenin nanoparticles in preparation of long-acting cefalonium preparation

Abstract

This invention discloses the application of naringenin nanoparticles in the preparation of long-acting ceftiofur formulations. Naringenin (5,7-dihydroxy-2(4-hydroxyphenyl)chroman-4-one) is a flavonoid. This invention prepares naringenin nanoparticles and combines them with ceftiofur. It was found that naringenin can effectively intervene in the pharmacokinetic process of ceftiofur, significantly prolonging its in vivo half-life, thereby extending the in vivo duration of action and improving its efficacy. Furthermore, this application method is simple, easy to operate, and inexpensive, making it suitable for large-scale promotion.

Description

Technical Field

[0001] This invention relates to the field of veterinary drug technology, specifically to the application of naringenin nanoparticles in the preparation of long-acting ceftiofur formulations. Background Technology

[0002] Ceftiofur is a cephalosporin antibiotic used in veterinary clinical practice, and is a broad-spectrum antibacterial drug. It has strong antibacterial activity against both Gram-positive and Gram-negative bacteria. Ceftiofur acts on transcriptional peptidase, blocking the synthesis of peptidoglycans, leading to bacterial cell wall loss and thus achieving bactericidal action. Ceftiofur has a stable β-lactam ring, which is not easily destroyed by drug-resistant bacteria, making it one of the most widely used therapeutic drugs in clinical practice. After intramuscular injection, ceftiofur sodium is rapidly absorbed and quickly generates its primary metabolite, defuranylceftiofur, in the plasma, further forming the active defuranylceftiofur cysteine ​​disulfide, which is then excreted in the urine and feces.

[0003] The short duration of effective blood drug concentration is one of the main reasons affecting the efficacy of ceftiofur. Therefore, long-acting formulations of this drug are prepared in clinical practice through various methods. However, although the effective blood drug concentration is improved after preparation, the complex preparation process, poor stability, increased cost, easy induction of drug tolerance, and increased risk of residues prevent the drug from exerting its best therapeutic potential. Therefore, it is essential to study a simple and economical method to improve the therapeutic effect of ceftiofur and prolong its duration of action in vivo. Summary of the Invention

[0004] The purpose of this invention is to address the aforementioned problems in the prior art and provide the application of naringenin nanoparticles in the preparation of long-acting ceftiofur formulations. This invention, through the use of naringenin nanoparticles, intervenes in the pharmacokinetic process of ceftiofur, prolongs the in vivo duration of action of ceftiofur, and enhances the therapeutic effect of ceftiofur.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: the application of naringenin nanoparticles in the preparation of long-acting ceftiofur formulations, wherein the naringenin nanoparticles are used to prolong the duration of action of ceftiofur in animals.

[0006] Preferably, naringenin nanoparticles are used in combination with ceftiofur; the application concentration of the naringenin nanoparticles is 1 mg / kg; and the application concentration of the ceftiofur is 5 mg / kg.

[0007] Preferably, the long-acting ceftiofur formulation is a powder for injection.

[0008] Preferably, the preparation method of the naringenin nanoparticles includes the following steps:

[0009] S1. Prepare a 1% sodium caseinate solution. After stirring until completely dissolved, incubate at 4°C overnight, adjust the pH to 3.5, and heat to 65°C to obtain a casein solution.

[0010] S2, naringenin is dissolved in 70% ethanol, added to the casein solution in S1, and then chitosan is added to make the final concentration 1 mg / ml; after stirring thoroughly, the mixed solution is transferred to a 4℃ water bath, and 1 mL of 1M tripotassium citrate, 6 mL of 0.2M K2HPO4, and 5 mL of 0.2M CaCl2 are added, maintaining the pH of the solution between 6.7 and 7.0 during the addition process;

[0011] S3. Add water to a volume of 100ml, stir and mix well, then centrifuge repeatedly for 3000g to remove unencapsulated naringin, and freeze-dry to obtain naringin nanoparticles.

[0012] Further preferred, the animals include pigs, ducks, and rabbits.

[0013] The beneficial effects of this invention: Naringenin (5,7-dihydroxy-2(4-hydroxyphenyl)chroman-4-one) is a flavonoid. This invention combines naringenin nanoparticles with ceftiofur, finding that the naringenin nanoparticles can effectively interfere with the pharmacokinetic process of ceftiofur, significantly prolonging its in vivo half-life, thereby extending the duration of action of ceftiofur in vivo and improving its efficacy. Furthermore, this application method is simple, easy to operate, and inexpensive, making it suitable for large-scale promotion. Attached Figure Description

[0014] Figure 1 The effect of naringenin nanoparticles on the blood concentration of ceftiofur in pigs;

[0015] Figure 2 The effect of naringenin nanoparticles on the blood concentration of ceftiofur in ducks;

[0016] Figure 3 The effect of naringenin nanoparticles on the blood concentration of ceftiofur in rabbits. Detailed Implementation

[0017] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0018] Example 1: Preparation of naringenin nanoparticles

[0019] First, prepare a 1% sodium caseinate solution. After complete dissolution by stirring, incubate at 4°C overnight to allow for full swelling. Adjust the pH of the casein solution to 3.5 after overnight incubation and heat to 65°C. Dissolve naringenin in 70% ethanol and add it to the casein solution, followed by chitosan to a final concentration of 1 mg / ml. After thorough stirring, transfer the solution to a 4°C water bath and add 1 mL of 1M tripotassium citrate, 6 mL of 0.2M K₂HPO₄, and 5 mL of 0.2M CaCl₂. During the addition process, maintain the pH of the solution between 6.7 and 7.0 using 1M HCl and 1M NaOH solutions. Finally, add water to a volume of 100 mL and stir until well mixed. Centrifuge the prepared solution repeatedly at 3000g to remove unencapsulated naringenin, and then freeze-dry to obtain the final product.

[0020] Example 2: Application of naringenin nanoparticles in the preparation of long-acting ceftiofur formulations

[0021] The naringenin nanoparticles used in this embodiment were prepared in Example 1.

[0022] 1. Pharmacokinetic study of enhanced ceftiofur in pigs

[0023] Healthy piglets were selected for the experiment and divided into two groups of six each: a control group (cefotiam, 5 mg / kg) and a combination group (1 mg / kg of naringenin nanoparticles mixed with 5 mg / kg of ceftiam). Following administration, 2 ml of blood was collected from the anterior vena cava at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours. Plasma was separated after anticoagulation with heparin sodium, and plasma concentrations of ceftiam were determined using high-performance liquid chromatography (HPLC). Drug-time curves were plotted, and pharmacokinetic parameters were statistically analyzed.

[0024] from Figure 1 It can be seen that naringenin nanoparticles significantly increased the blood concentration of ceftiofur in pigs and altered the in vivo duration of action of ceftiofur. As shown in Table 1, naringenin significantly increased the area under the curve (AUC), residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial for maintaining effective blood concentration.

[0025] Table 1. Effects of naringenin nanoparticles on pharmacokinetic parameters of ceftiofur in pigs. 2.

[0027] 3. Pharmacokinetic Study of Enhanced Ceftiofur in Ducks

[0028] Healthy Tianfu meat ducks were selected for the experiment and divided into two groups of 6 ducks each: a control group (cefotiam group, 5 mg / kg) and a combination group (1 mg / kg of naringenin nanoparticles mixed with 5 mg / kg of ceftiam). After administration, 2 ml of blood was collected from the anterior vena cava at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours. Plasma was separated after anticoagulation with heparin sodium, and the plasma concentration of ceftiam was detected using high-performance liquid chromatography (HPLC). Drug-time curves were plotted, and pharmacokinetic parameters were statistically analyzed.

[0029] from Figure 2 It can be seen that naringenin nanoparticles significantly increased the blood concentration of ceftiofur in ducks and altered the in vivo duration of action of ceftiofur. As shown in Table 2, naringenin significantly increased the area under the curve (AUC), residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial for maintaining effective blood concentration.

[0030] Table 2. Effects of naringenin nanoparticles on pharmacokinetic parameters of ceftiofur in ducks.

[0031]

[0032] 4. Pharmacokinetic study of enhanced ceftiofur in rabbits

[0033] Healthy New Zealand rabbits were selected for the experiment and divided into two groups of six each: a control group (cefotiam, 5 mg / kg) and a combination group (1 mg / kg of naringenin nanoparticles mixed with 5 mg / kg of ceftiam). Following administration, 2 ml of blood was collected from the anterior vena cava at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours. Plasma was separated after anticoagulation with heparin sodium, and plasma concentrations of ceftiam were determined using high-performance liquid chromatography (HPLC). Drug-time curves were plotted, and pharmacokinetic parameters were statistically analyzed.

[0034] from Figure 3 It can be seen that naringenin nanoparticles significantly increased the blood concentration of ceftiofur in rabbits and altered the in vivo duration of action of ceftiofur. As shown in Table 3, naringenin significantly increased the area under the curve (AUC), residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial for maintaining effective blood concentration.

[0035] Table 3 Effects of naringenin nanoparticles on pharmacokinetic parameters of ceftiofur in rabbits

[0036]

[0037] In summary, the present invention combines naringenin nanoparticles with ceftiofur, which can significantly increase the blood concentration of ceftiofur in animals, prolong the maintenance time of effective blood concentration, slow down the elimination of ceftiofur, and thus improve the efficacy of the drug.

[0038] The specification and drawings of this invention are intended to be illustrative rather than restrictive. Based on this invention, those skilled in the art can make substitutions and modifications to some of the technical features without creative effort, and all such modifications are within the scope of protection of this invention.

Claims

1. Use of naringenin nanoparticles in the preparation of a long-acting formulation of ceftiofur, characterized in that, Naringenin nanoparticles are used in combination with ceftiofur, wherein the naringenin nanoparticles are used to prolong the duration of action of ceftiofur in animals; the preparation method of the naringenin nanoparticles includes the following steps: S1. Prepare a 1% sodium caseinate solution. After stirring until completely dissolved, incubate at 4°C overnight, adjust the pH to 3.5, and heat to 65°C to obtain a casein solution. S2, naringenin is dissolved in 70% ethanol, added to the casein solution in S1, and then chitosan is added to make the final concentration 1 mg / ml; after stirring thoroughly, the mixed solution is transferred to a 4℃ water bath, and 1 mL of 1M tripotassium citrate, 6 mL of 0.2M K2HPO4, and 5 mL of 0.2MCaCl2 are added, maintaining the pH of the solution between 6.7 and 7.0 during the addition process; S3. Add water to a volume of 100ml, stir and mix well, then centrifuge repeatedly for 3000g to remove unencapsulated naringin, and freeze-dry to obtain naringin nanoparticles.

2. Use according to claim 1, characterized in that, The application concentration of the naringenin nanoparticles is 1 mg / kg; the application concentration of the ceftiofur is 5 mg / kg.

3. Use according to claim 1, characterized in that, The long-acting ceftiofur formulation is a powder for injection.

4. Use according to claim 1, characterized in that, The animals mentioned include pigs, ducks, and rabbits.

Images