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

Abstract

The invention discloses an application of naringenin nanoparticles in preparation of a ceftiofur long-acting preparation. The invention discloses naringenin (5, 7-dihydroxy-2 (4-hydroxyphenyl) chroman-4-ketone), which is a flavonoid substance. The naringenin nanoparticles are prepared, the naringenin nanoparticles are mixed with the ceftiofur for combined use, and it is found that the naringenin can effectively intervene in the pharmacokinetic process of the ceftiofur and significantly prolong the in-vivo half-life period of the ceftiofur, so that the in-vivo action time of the ceftiofur is prolonged, and the curative effect of the ceftiofur is improved. Moreover, the application method is simple, easy to operate, low in cost and suitable for large-scale popularization.

Description

Technical Field

[0001] The invention relates to the technical field of veterinary drugs, and in particular to application of naringenin nanoparticles in the preparation of a long-acting ceftiofur preparation. Background Art

[0002] Ceftiofur is a cephalosporin antibiotic used in veterinary clinics and is a broad-spectrum antibacterial drug. It has strong antibacterial effects on both Gram-positive and Gram-negative bacteria. Ceftiofur acts on transcript peptidase to block the synthesis of mucopeptides, causing bacterial cell wall loss and achieving bactericidal effects. Ceftiofur has a stable β-lactam ring and is not easily destroyed by drug-resistant bacteria. It is one of the most widely used therapeutic drugs in clinical practice. Ceftiofur sodium is rapidly absorbed after intramuscular injection and quickly generates a primary metabolite in plasma, defuroyl ceftiofur, which further forms active defuroyl ceftiofur cysteine ​​disulfide and is excreted through the urinary tract and feces.

[0003] The short maintenance time of effective blood drug concentration is one of the main reasons affecting the effect of ceftiofur. Therefore, long-acting preparations of the drug are prepared in clinical practice in various ways. However, although the effective blood drug concentration is improved after preparation, the drug cannot exert its best therapeutic potential due to factors such as complex preparation process, poor stability, increased cost, easy induction of drug tolerance, and increased residual risk. Therefore, it is necessary to study a simple and economical method to improve the therapeutic effect of ceftiofur and prolong the in vivo action time. Summary of the invention

[0004] The purpose of the present invention is to solve the above problems existing in the prior art and provide the use of naringenin nanoparticles in the preparation of a long-acting preparation of ceftiofur. The present invention intervenes in the pharmacokinetic process of ceftiofur, prolongs the in vivo action time of ceftiofur, and enhances the therapeutic effect of ceftiofur through the use of naringenin nanoparticles.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows: application of naringenin nanoparticles in the preparation of a long-acting preparation of ceftiofur, wherein the naringenin nanoparticles are used to prolong the action time of ceftiofur in animals.

[0006] Preferably, naringenin nanoparticles are mixed with ceftiofur for combined use; the application concentration of the naringenin nanoparticles is 1 mg / kg; the application concentration of the ceftiofur is 5 mg / kg.

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

[0008] Preferably, the method for preparing the naringenin nanoparticles comprises the following steps:

[0009] S1. Prepare a 1% sodium caseinate solution, stir to completely dissolve, adjust the pH to 3.5 after overnight at 4°C, and heat to 65°C to obtain a casein solution;

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

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

[0012] More preferably, the animals include pigs, ducks and rabbits.

[0013] The beneficial effects of the present invention: Naringenin (5,7-dihydroxy-2 (4-hydroxyphenyl) chroman-4-one) is a flavonoid. The present invention uses naringenin nanoparticles mixed with ceftiofur, and finds that naringenin nanoparticles can effectively intervene in the pharmacokinetic process of ceftiofur, significantly prolong the in vivo half-life of ceftiofur, thereby prolonging the in vivo action time of ceftiofur and improving the efficacy of ceftiofur. In addition, the application method is simple, easy to operate, and low in cost, and is suitable for large-scale promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[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 DESCRIPTION

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

[0018] Example 1 Preparation of Naringin Nanoparticles

[0019] First, prepare a 1% sodium caseinate solution, stir until completely dissolved, and incubate at 4°C overnight to fully swell. After overnight, adjust the pH of casein to 3.5 and heat to 65°C. Dissolve naringenin in 70% ethanol, add casein solution, and then add chitosan to a final concentration of 1mg / ml. After sufficient stirring, transfer the solution to a 4°C water bath, add 1mL of 1M tripotassium citrate, 6mL of 0.2M K2HPO4, and 5mL of 0.2M CaCl2. During the addition process, use 1M HCL and 1M NaOH solutions to maintain the pH of the solution between 6.7-7.0. Finally, add water to a volume of 100ml and stir to mix. The prepared solution is repeatedly centrifuged at 3000g to remove unencapsulated naringenin, and then freeze-dried.

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

[0021] The naringenin nanoparticles used in this example 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, 6 in each group, namely the control group (ceftiofur group, 5 mg / kg) and the combined group (naringenin nanoparticles 1 mg / kg mixed with ceftiofur 5 mg / kg). After medication, 2 ml of anterior vena cava blood was collected at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours, and plasma was separated after anticoagulation with heparin sodium. The blood concentration of ceftiofur in plasma was detected by high-efficiency chromatography, and the drug-time curve was drawn and the 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 changed the in vivo action time of ceftiofur. As shown in Table 1, naringenin significantly increased the area under the drug-time curve (AUC), in vivo residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial to the maintenance of effective blood concentration.

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

[0026] parameter unit Rabbit control group Rabbit Enhanced Group AUC(0-t) mg / L*h 74.381±1.191 113.802±2.658* AUC(0-∞) mg / L*h 76.323±0.703 117.137±3.262* MRT(0-∞) h 5.986±0.089 9.728±0.327* t1 / 2z h 4.558±0.814 6.957±0.556* CL L / h / kg 0.066±0.001 0.043±0.001* V L / kg 0.431±0.077 0.428±0.027 C mg / L 0.284±0.081 0.328±0.066*

[0027] 2. Pharmacokinetic study of enhanced ceftiofur in ducks

[0028] Healthy Tianfu ducks were selected for the experiment and divided into two groups, 6 in each group, namely the control group (ceftiofur group, 5 mg / kg) and the combined group (naringenin nanoparticles 1 mg / kg mixed with ceftiofur 5 mg / kg). After medication, 2 ml of anterior vena cava blood was collected at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours, and plasma was separated after anticoagulation with heparin sodium. The blood concentration of ceftiofur in plasma was detected by high-efficiency chromatography, and the drug-time curve was drawn and the 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 changed the in vivo action time of ceftiofur. As shown in Table 2, naringenin significantly increased the area under the drug-time curve (AUC), in vivo residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial to the maintenance of effective blood concentration.

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

[0031]

[0032]

[0033] 3. Pharmacokinetic study of enhanced ceftiofur in rabbits

[0034] Healthy New Zealand rabbits were selected for the experiment and divided into two groups, 6 in each group, namely the control group (ceftiofur group, 5 mg / kg) and the combined group (naringenin nanoparticles 1 mg / kg mixed with ceftiofur 5 mg / kg). After medication, 2 ml of anterior vena cava blood was collected at 0.1, 0.2, 0.4, 0.7, 1, 1.5, 2, 3, 4, 5, 8, 12, 18, 24, and 36 hours, and plasma was separated after anticoagulation with heparin sodium. The blood concentration of ceftiofur in plasma was detected by high-efficiency chromatography, and the drug-time curve was drawn and the pharmacokinetic parameters were statistically analyzed.

[0035] from Figure 3 It can be seen that naringenin nanoparticles significantly increased the blood concentration of ceftiofur in rabbits and changed the in vivo action time of ceftiofur. As shown in Table 3, naringenin significantly increased the area under the drug-time curve (AUC), in vivo residence time (MRT), and half-life (t1 / 2z) of ceftiofur, which is beneficial to the maintenance of effective blood concentration.

[0036] Table 3 Effect of naringenin nanoparticles on the pharmacokinetic parameters of ceftiofur in rabbits

[0037] parameter unit Rabbit control group Rabbit Enhanced Group AUC(0-t) mg / L*h 74.381±1.191 113.802±2.658* AUC(0-∞) mg / L*h 76.323±0.703 117.137±3.262* MRT(0-∞) h 5.986±0.089 9.728±0.327* t1 / 2z h 4.558±0.814 6.957±0.556* CL L / h / kg 0.066±0.001 0.043±0.001* V L / kg 0.431±0.077 0.428±0.027 C mg / L 0.284±0.081 0.328±0.066*

[0038] In summary, the combined use of naringenin nanoparticles and ceftiofur in the present invention can significantly increase the blood concentration of ceftiofur in animals, prolong the maintenance time of the effective blood concentration, slow down the elimination of ceftiofur, and is conducive to improving the effect of the drug.

[0039] The specification and drawings of the present invention are considered to be illustrative rather than restrictive. On the basis of the present invention, those skilled in the art can make some substitutions and modifications to some of the technical features according to the disclosed technical content without creative labor, all of which are within the protection scope of the present invention.

Claims

1. Application of naringenin nanoparticles in the preparation of a long-acting ceftiofur preparation, characterized in that: The naringenin nanoparticles are used to prolong the action time of ceftiofur in animals.

2. The use according to claim 1, characterized in that: The naringenin nanoparticles are mixed with ceftiofur for joint use; the application concentration of the naringenin nanoparticles is 1 mg / kg; the application concentration of the ceftiofur is 5 mg / kg.

3. The use according to claim 1, characterized in that: The long-acting preparation of ceftiofur is a powder injection.

4. The use according to claim 1, characterized in that: The preparation method of the naringenin nanoparticles comprises the following steps: S1. Prepare a 1% sodium caseinate solution, stir to completely dissolve, adjust the pH to 3.5 after overnight at 4°C, and heat to 65°C to obtain a casein solution; S2, naringenin were dissolved in 70% ethanol, the casein solution in S1 was added, and then chitosan was added to make the final concentration 1mg / ml; after sufficient stirring, the mixed solution was transferred to a 4°C water bath, 1mL 1M tripotassium citrate, 6mL 0.2M K2HPO4, and 5mL 0.2M CaCl2 were added, and the pH of the solution was maintained between 6.7-7.0 during the addition process; S3. Add water to a volume of 100 ml, stir and mix, then centrifuge repeatedly at 3000 g to remove unencapsulated naringenin, and freeze-dry to obtain naringenin nanoparticles.

5. The use according to claim 1, characterized in that: The animals include pigs, ducks and rabbits.

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