A long-acting telithromycin injection and a method for preparing the same

Abstract

The present application relates to long-acting telithromycin injection technical field, especially to a kind of long-acting telithromycin injection and preparation method thereof, the present application is prepared into inclusion compound by telithromycin, then with emulsifying agent, coemulsifier, isotonic agent, complexing agent, tryptophan and injection oil and injection water are prepared into long-acting telithromycin injection, the introduction of emulsifying agent, coemulsifier and complexing agent can effectively improve the stability of injection, the introduction of tryptophan can control the release rate and release amount of telithromycin, so that the long-acting telithromycin injection prepared finally has good security and stability, and has the advantages of long-acting and controllable.

Description

Technical Field

[0001] This invention relates to the field of long-acting tylosin injection technology, and in particular to a long-acting tylosin injection and its preparation method. Background Technology

[0002] Tilatomycin is a broad-spectrum antibiotic highly effective against many pathogens causing respiratory diseases in pigs and cattle, such as hemolytic Pasteurella multocida, Pasteurella hemorrhagicis, Actinobacillus pleuropneumoniae, Mycoplasma, Haemophilus parasuis, Bordetella bronchiseptica, and Bovine infectious keratoconjunctivitis caused by Moraxella vesiculosus. It possesses numerous advantages, including a broad antibacterial spectrum, high activity, low clinical dosage, low toxicity, low residue, non-teratogenicity, and the ability to complete the entire treatment process with a single dose. Therefore, tilatomycin has attracted widespread attention worldwide since its market launch. Commercially available tilatomycin products are injection solutions, which are water-miscible. Most of the patented technologies disclose the preparation of tilatomycin injections using organic solvents / water co-solvents and acidic conditions (CN1429232A, CN101422479A, CN102048747A, CN101416977A). Currently available tylosin injections and the formulations disclosed in the above patents share a common drawback: they are highly irritating, often causing temporary pain and swelling at the injection site during injection.

[0003] Tylopycin is poorly soluble in water but readily soluble in acidic solutions. Most commercially available long-acting tylopycin injections are solution-type injectable formulations prepared under acidic conditions using organic solvents / water co-solvents. These solutions are acidic and highly irritating, often causing injection pain and swelling at the injection site. Oil-based tylopycin injections, compared to solution-type injections, have the disadvantage of slow onset of action, making them unsuitable for treating acute severe infections in animals. Furthermore, while the addition of antioxidants to long-acting tylopycin injection formulations improves drug stability, it reduces the safety of administration.

[0004] Therefore, there is an urgent need to provide a long-acting tylosin injection that is safe, fast-acting, highly soluble, stable, and has a controllable release rate. Summary of the Invention

[0005] In order to solve the problems of slow onset of action, poor solubility, poor stability and poor controllability of existing long-acting tylosin injections, the present invention aims to provide a long-acting tylosin injection and its preparation method to solve the problems existing in the prior art.

[0006] To achieve the above objectives, the present invention provides a long-acting tylosin injection, characterized in that it comprises the following components: tylosin-β-cyclodextrin inclusion complex, emulsifier, co-emulsifier, tryptophan, isotonic agent, complexing agent, injection oil, and water for injection; each 100 mL of the long-acting tylosin injection contains:

[0007] Tylamycin-β-cyclodextrin inclusion complex 20g

[0008] Emulsifier 1.5-3g

[0009] Co-emulsifier 0.1-1g

[0010] 2-3g of isotonic agent

[0011] 4-8g of tryptophan

[0012] Complexing agent 0.01-1g

[0013] 10-30g of oil for injection

[0014] The remainder is water for injection.

[0015] Preferably, in one technical solution, the β-cyclodextrin is selected from one of: hydroxypropyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin or glucosyl-β-cyclodextrin;

[0016] Preferably, in one technical solution, the tylosin-β-cyclodextrin inclusion complex is selected from one of the following: tylosin hydroxypropyl-β-cyclodextrin inclusion complex, tylosin hydroxyethyl-β-cyclodextrin inclusion complex, tylosin sulfobutyl ether-β-cyclodextrin inclusion complex, or tylosin glucosyl-β-cyclodextrin inclusion complex; the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1-4; preferably, the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1-2; more preferably, the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1;

[0017] Preferably, in one technical solution, the amount of tylosin-β-cyclodextrin inclusion complex in the long-acting tylosin injection is 200 g / L;

[0018] Preferably, in one technical solution, the dosage of tylosin in the long-acting tylosin injection is 100 g / L;

[0019] Preferably, in one embodiment, the emulsifier is selected from one or a combination of two of soybean lecithin or egg yolk lecithin, and the amount of emulsifier used in the long-acting tylosin injection is 10-30 g / L; preferably, the amount of emulsifier used in the long-acting tylosin injection is 15-30 g / L; more preferably, the amount of emulsifier used in the long-acting tylosin injection is 15 g / L.

[0020] Preferably, in one embodiment, the co-emulsifier is selected from one or a combination of two of PEG400 or PEG600; the amount of co-emulsifier in the long-acting tylosin injection is 1-10 g / L; preferably, the amount of co-emulsifier in the long-acting tylosin injection is 6-10 g / L.

[0021] Preferably, in one embodiment, the isotonic agent is selected from one or more combinations of glycerol, xylitol, or glucose, and the amount of isotonic agent used in the long-acting tylosin injection is 20-30 g / L; preferably 20 g / L, 26 g / L, or 30 g / L.

[0022] Preferably, in one technical solution, the amount of tryptophan in the long-acting tylosin injection is 40-80 g / L; more preferably, it is 50-80 g / L; and even more preferably, it is 40-60 g / L.

[0023] Preferably, in one embodiment, the complexing agent is selected from disodium ethylenediaminetetraacetate (EDTA), and the amount of EDTA in the long-acting tylosin injection is 0.1-10 g / L; preferably, the amount of EDTA in the long-acting tylosin injection is 0.1-0.5 g / L.

[0024] Preferably, in one embodiment, the oil for injection is selected from one or more of soybean oil, peanut oil, tea oil, sesame oil or olive oil, and the amount of oil for injection in the long-acting tylosin injection is 100-300 g / L; more preferably, the amount of oil for injection in the long-acting tylosin injection is 150-200 g / L.

[0025] This invention also provides a method for preparing a long-acting tylosin injection, comprising the following steps:

[0026] The specific steps are as follows:

[0027] 1) Preparation of tylosin-β-cyclodextrin inclusion complex:

[0028] A certain amount of β-cyclodextrin was dissolved in an appropriate amount of distilled water, heated to 50-60℃, and stirred until completely dissolved to obtain solution I. A certain amount of tylosin was dissolved in anhydrous ethanol to obtain solution II. Solution II was slowly added dropwise to solution I under constant stirring. Inclusion was carried out in a constant temperature water bath at 50℃. After the addition was completed, stirring was continued to ensure full inclusion. The mixture was then refrigerated and pre-frozen, and freeze-dried in a freeze dryer to obtain the tylosin hydroxypropyl-β-cyclodextrin inclusion complex. The mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex was 1:1-4.

[0029] 2) Under nitrogen protection at 50°C, the emulsifier is completely dissolved in a certain amount of injection oil. The mixture is stirred until it is completely dissolved to obtain the oil phase.

[0030] 3) Take a certain amount of the tylosin-β-cyclodextrin inclusion complex, isotonic agent, tryptophan, co-emulsifier, and complexing agent prepared in step 1) and dissolve them in a certain amount of water for injection. Stir until completely dissolved to obtain an aqueous phase.

[0031] 4) Mix the above oil phase and water phase, bring the volume to 100 mL, and then transfer it to a high-speed shear emulsifier. Stir and shear at high speed to obtain the primary emulsion.

[0032] 5) The prepared primary emulsion is then subjected to high-pressure homogenization, and the homogenized emulsion is filtered through a 0.22μm microporous membrane, filled with nitrogen, sealed, and sterilized at 115℃ for 30 minutes to obtain the final product.

[0033] Preferably, the heating temperature in step 1) is 50-60°C, more preferably, the temperature is 50°C;

[0034] Preferably, the β-cyclodextrin in step 1) is: hydroxypropyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, or glucosyl-β-cyclodextrin;

[0035] Preferably, the mass ratio of β-cyclodextrin to distilled water in step 1) is 1:10-30; more preferably, it is 1:15-20.

[0036] Preferably, the encapsulation time in step 1) is 30-120 min; more preferably, the encapsulation time is 30-60 min.

[0037] Preferably, the tylosin-β-cyclodextrin inclusion complex in step 1) is selected from: tylosin hydroxypropyl-β-cyclodextrin inclusion complex, tylosin hydroxyethyl-β-cyclodextrin inclusion complex, tylosin sulfobutyl ether-β-cyclodextrin inclusion complex, and tylosin glucosyl-β-cyclodextrin inclusion complex; the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1-4; preferably, the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1-2; more preferably, the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1;

[0038] Preferably, the oil used for injection in step 2) is selected from one or more of soybean oil, peanut oil, tea oil, sesame oil or olive oil, and the amount of the oil used for injection in step 2) relative to the long-acting tylosin injection is 100-300 g / L; more preferably, it is 150-200 g / L.

[0039] Preferably, the emulsifier in step 2) is selected from one or a combination of two of soybean lecithin or egg yolk lecithin, and the amount of the emulsifier relative to the long-acting tylosin injection is 10-30 g / L; the preferred amount is 15-30 g / L; and the more preferred amount is 15 g / L.

[0040] Preferably, in step 3), the mass of tylosin in the tylosin-β-cyclodextrin inclusion complex is 10g;

[0041] Preferably, in step 3), the co-emulsifier is selected from one or a combination of two of PEG400 or PEG600; the amount of co-emulsifier in step 3) relative to the long-acting tylosin injection is 1-10 g / L; preferably, the amount of co-emulsifier is 6-10 g / L.

[0042] Preferably, the isotonic agent in step 3) is selected from one or more of glycerol, xylitol or glucose, and the amount of the isotonic agent in step 3) relative to the long-acting tylosin injection is 20-30 g / L; preferably 20 g / L, 26 g / L or 30 g / L.

[0043] Preferably, the complexing agent in step 3) is selected from disodium ethylenediaminetetraacetate; the amount of disodium ethylenediaminetetraacetate in step 3) relative to long-acting tylosin injection is 0.1-10 g / L; preferably, the amount of disodium ethylenediaminetetraacetate is 0.1-0.5 g / L.

[0044] Preferably, in step 3), the amount of tryptophan relative to long-acting tylosin injection is 40-80 g / L; more preferably, it is 50-80 g / L; and even more preferably, it is 40-60 g / L.

[0045] Preferably, the rotation speed of the high-speed stirring and shearing in step 4) is 10000 r; the high-speed stirring and shearing time is 3 min-10 min.

[0046] Preferably, in step 5), the high-pressure homogenization temperature is 50-60℃, the pressure is 20-30MPa, the time is 30-60min, and the number of cycles is 3-5.

[0047] The beneficial effects of this invention are as follows:

[0048] 1) The injection solution prepared by this invention is non-irritating, safe, fast-acting, has good solubility, high stability, and good controllability, making it suitable for industrial production. Detailed Implementation

[0049] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0050] The present invention will be further explained below with reference to specific embodiments.

[0051] Example 1

[0052] Prescription for long-acting tylosin injection (100mL):

[0053] 20g of tylosin hydroxypropyl-β-cyclodextrin inclusion complex

[0054] 15g of soybean oil for injection

[0055] 1.5g of soybean lecithin

[0056] PEG400 0.6g

[0057] 2.6g of glycerin

[0058] 5g of tryptophan

[0059] Disodium ethylenediaminetetraacetate 0.02g

[0060] The remainder is water for injection.

[0061] 1) Preparation of tylosin hydroxypropyl-β-cyclodextrin inclusion complex:

[0062] 20g of hydroxypropyl-β-cyclodextrin was dissolved in 300mL of distilled water, heated to 50℃, and stirred until completely dissolved to obtain solution I. 20g of tylosin was dissolved in 30mL of anhydrous ethanol to obtain solution II. Solution II was slowly added dropwise to solution I under constant stirring. Inclusion was carried out in a constant temperature water bath at 50℃. After the addition was completed, stirring was continued for 2 hours to ensure full inclusion. The mixture was then pre-frozen in a refrigerated freeze dryer and freeze-dried to obtain the tylosin hydroxypropyl-β-cyclodextrin inclusion complex. The mass ratio of tylosin to hydroxypropyl-β-cyclodextrin in the tylosin hydroxypropyl-β-cyclodextrin inclusion complex was 1:1.

[0063] 2) Under nitrogen protection at 50℃, 1.5g of soybean lecithin was completely dissolved in 15g of soybean oil for injection. The mixture was stirred until it was completely dissolved to obtain the oil phase.

[0064] 3) Take 20g of the tylosin hydroxypropyl-β-cyclodextrin inclusion complex prepared in step 1), 2.6g of glycerol, 5g of tryptophan, and 0.02g of disodium ethylenediaminetetraacetate and dissolve them in 70mL of water for injection. Stir until completely dissolved to obtain the aqueous phase.

[0065] 4) Mix the above oil phase and water phase, bring the volume to 100 mL, and then transfer it to a high-speed shear emulsifier. Stir and shear at high speed (10000 r, 3 min) to obtain the primary emulsion.

[0066] 5) The prepared primary emulsion is then subjected to high-pressure homogenization at a temperature of 50°C, a pressure of 20 MPa, a time of 30 min, and a cycle of 3 times. The homogenized emulsion is then filtered through a 0.22 μm microporous membrane, filled with nitrogen, sealed, and sterilized at 115°C for 30 min to obtain the final product.

[0067] Examples 2-4

[0068] Examples 2-4 were prepared according to the method of Example 1, and the specific components and amounts are shown in Table 1.

[0069] Table 1

[0070]

[0071]

[0072] Comparative Examples 1-3

[0073] Comparative Examples 1-3 were prepared according to the method of Example 1, and the specific components and amounts are shown in Table 2.

[0074] Table 2

[0075]

[0076] "-" indicates that no addition was made.

[0077] Example 5

[0078] The samples prepared in Examples 1-2 and Comparative Examples 2-3 were subjected to accelerated testing and long-term stability studies. The study conditions were as follows: 1) Accelerated testing was conducted for 6 months at a high temperature of 40±2℃ and a relative humidity of 75%±5%, with samples taken once at months 1, 2, 3, and 6. The results are shown in Table 3. 2) Long-term stability testing was conducted for 12 months at a high temperature of 25±2℃ and a relative humidity of 60%±5%, with samples taken once at months 3, 6, 9, and 12. The results are shown in Table 4.

[0079] Table 3 Results of Accelerated Experiments

[0080]

[0081]

[0082] Table 4 Results of Long-Term Stability Test

[0083]

[0084]

[0085] As can be seen from the results in Tables 3 and 4, when the amount of co-emulsifier is reduced (Comparative Example 2), the stability of the prepared injection solution decreases. Without the addition of a complexing agent (Comparative Example 3), the stability of the prepared injection solution also decreases. However, the stability of the injection solution of this application is significantly improved after adding a certain amount of co-emulsifier and complexing agent. It is evident that the injection solution prepared by this invention has excellent stability.

[0086] Example 6

[0087] Blood drug concentration determination of long-acting tylosin injection

[0088] The long-acting tylosin injection prepared in Examples 1-2 was used as the drug and compared with the tylosin prepared in Comparative Example 1. Fifteen healthy pigs, weighing 40-50 kg, were selected and divided into three groups of five pigs each. Each group received an intramuscular injection in the neck at a dose of 2.5 mg / kg. Blood samples were collected regularly to detect the tylosin content (ng / ml) in the plasma. Details are shown in Table 5 below.

[0089] Table 5

[0090]

[0091] As shown in Table 5, the amount of tryptophan added is related to the release rate and amount of tylosin. When the amount of tryptophan added is low (e.g., 2g / 100mL), the release rate of tylosin is slow and the total amount of tylosin released is relatively low. However, when the amount of tryptophan added is high (e.g., 5g / 100mL), the release rate of tylosin is significantly faster and the total amount of tylosin released is also greater. This may be related to the fact that tryptophan can competitively bind to hydroxypropyl-β-cyclodextrin with tylosin. Therefore, the addition of tryptophan to the formulation in this invention can control the release rate and amount of tylosin.

[0092] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A long-acting tylosin injection, characterized in that, It consists of the following components: tylosin-β-cyclodextrin inclusion complex, emulsifier, co-emulsifier, isotonic agent, tryptophan, complexing agent, oil for injection and water for injection; The tylosin-β-cyclodextrin inclusion complex is selected from: tylosin hydroxypropyl-β-cyclodextrin inclusion complex; the mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex is 1:1; the amount of tylosin hydroxypropyl-β-cyclodextrin inclusion complex is 200 g / L; The emulsifier is selected from soybean lecithin; the amount of emulsifier used in the long-acting tylosin injection is 15 g / L; The co-emulsifier is selected from PEG400; the amount of co-emulsifier in the long-acting tylosin injection is 6-10 g / L; The isotonic agent is selected from glycerin; the dosage of the isotonic agent in the long-acting tylosin injection is 26 g / L; The dosage of tryptophan in the long-acting tylosin injection is 40-60 g / L; The complexing agent is selected from disodium ethylenediaminetetraacetate, and the dosage of disodium ethylenediaminetetraacetate in the long-acting tylosin injection is 0.1-0.5 g / L; The oil used for injection is selected from soybean oil; the amount of oil used for injection in the long-acting tylosin injection is 150 g / L.

2. A method for preparing the long-acting tylosin injection according to claim 1, comprising the following steps: 1) Preparation of tylosin-β-cyclodextrin inclusion complex: A certain amount of β-cyclodextrin was dissolved in an appropriate amount of distilled water, heated to 50-60℃, and stirred until completely dissolved to obtain solution I. A certain amount of tylosin was dissolved in anhydrous ethanol to obtain solution II. Solution II was slowly added dropwise to solution I under constant stirring. Inclusion was carried out in a constant temperature water bath at 50℃. After the addition was completed, stirring was continued to ensure full inclusion. The mixture was then pre-frozen in a refrigerated freeze dryer and freeze-dried to obtain the tylosin hydroxypropyl-β-cyclodextrin inclusion complex. The mass ratio of tylosin to β-cyclodextrin in the tylosin-β-cyclodextrin inclusion complex was 1:

1. 2) Under nitrogen protection at 50°C, the emulsifier is completely dissolved in a certain amount of injection oil. The mixture is stirred until it is completely dissolved to obtain the oil phase. 3) Take a certain amount of the tylosin-β-cyclodextrin inclusion complex, isotonic agent, tryptophan, co-emulsifier, and complexing agent prepared in step 1) and dissolve them in a certain amount of water for injection. Stir until completely dissolved to obtain an aqueous phase. 4) Mix the above oil phase and water phase, bring the volume to 100 mL, and then transfer it to a high-speed shear emulsifier. Stir and shear at high speed to obtain the primary emulsion. 5) The prepared primary emulsion is then subjected to high-pressure homogenization, and the homogenized emulsion is filtered through a 0.22μm microporous membrane, filled with nitrogen, sealed, and sterilized at 115℃ for 30 minutes to obtain the final product.

3. The method for preparing long-acting tylosin injection according to claim 2, characterized in that, The β-cyclodextrin in step 1) is selected from: hydroxypropyl-β-cyclodextrin; In step 1), the mass ratio of β-cyclodextrin to distilled water is 1:15-20; The encapsulation time in step 1) is 30-60 minutes; In step 1), the tylosin-β-cyclodextrin inclusion complex is selected from: tylosin-hydroxypropyl-β-cyclodextrin inclusion complex.

4. The method for preparing long-acting tylosin injection according to claim 2, characterized in that, In step 4), the rotation speed of the high-speed stirring and shearing is 10000 r; the high-speed stirring and shearing time is 3 min to 10 min.

5. The method for preparing long-acting tylosin injection according to claim 2, characterized in that, In step 5), the temperature of high-pressure homogenization is 50-60℃, the pressure is 20-30MPa, the time is 30-60min, and the number of cycles is 3-5.

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