CN116211800A - Non-prednisone drop and preparation method and application thereof - Google Patents

Abstract

The invention relates to the technical field of medicines, in particular to a non-prednisone drop and a preparation method and application thereof. The non-prednisone drops comprise: a non-splash Luo Niwei ball and a vehicle; the non-prednisone Luo Niwei spheres comprise non-prednisone and MPEG-PLGA; the solvent comprises a solvent and an antioxidant. The preparation method of the non-prednisone Luo Niwei ball comprises the following steps: (1) Dissolving non-prednisone and MPEG-PLGA in acetone to obtain an oil phase A; dissolving polyvinyl alcohol in water to obtain a water phase B, injecting the oil phase A into the water phase B under high-speed shearing, and mixing and emulsifying the two phases to form O/W emulsion; (2) Drying the O/W emulsion at low temperature under reduced pressure, removing acetone, solidifying the microspheres, and filtering to remove water in the system after solidification to obtain solidified microspheres; (3) Dispersing the solidified microspheres in polyvinylpyrrolidone aqueous solution, and freeze-drying to obtain the non-prednisolone Luo Niwei spheres. The non-prednisone drop provided by the invention has longer killing time and higher safety.

Description

Non-prednisone drop and preparation method and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a non-prednisone drop and a preparation method and application thereof.

Background

The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.

Non-prednisone (Fipronil), also known as Fipronil, chemical name: the 5-amino-3-cyano-1- (2, 6-dichloro-4-trifluoromethylphenyl) -4-trifluoromethyl sulfinyl pyrazole is a phenylpyrazole pesticide, and has the action mechanism that the phenylpyrazole pesticide is combined with gamma-aminobutyric acid (GABA) receptors on insect central nervous cell membranes to close chloride ion channels of nerve cells and interfere normal functions of the central nervous system, so that nerves and muscles of insects are in an excited state for a long time, and finally the insects die. It mainly acts on the insects by stomach toxicity and contact killing, and also has a certain systemic conduction effect. Compared with traditional pesticides (such as benzimidazole, carbamate and the like), non-prednisone is safer for non-target animals, but has certain toxicity for human bodies and animals.

Currently, the flean of the French Mei Rui company occupies a large share in the domestic market of insect repellent products, the product comprises drops (the effective components are non-prednisone and methoprene) and spray (the effective components are non-prednisone), and is used for expelling and killing adult fleas, flea eggs, larvae and ticks on the surfaces of dogs and cats. Wherein the spray is suitable for treating fleas and dogs and cats with severe skin infections, and the drops are used for periodic prevention. After the product is applied to the skin surface of animals, the active ingredients are diffused to the whole body surface along with sebum and stored in sebaceous glands, and after that, the medicament is continuously supplemented to the whole body surface along with grease secreted by the sebaceous glands along with hair follicles, and the expelling and killing efficacy is about one month.

However, the inventor found that for dogs and cats that need to be exposed to the complex environment frequently, the insect repellent needs to be performed periodically, and the animal's compliance when the agent is applied is poor in consideration of the high price of the insect repellent product, and the expelling efficacy for one month obviously cannot meet the user's needs. In addition, after the Fulaien drops are applied, part of the medicine still enters the circulatory system of the animal through skin, has potential toxic and side effects, and has the phenomena of skin redness, dehairing and the like after the animals are applied.

Disclosure of Invention

In order to solve the technical problems of short expelling and killing time and low safety of an insect expelling product in the prior art, the invention discloses a non-prednisone drop with longer expelling and killing time and higher safety, a preparation method and application thereof, wherein the drop is prepared by using methoxy polyethylene glycol-poly (D, L-lactide-co-glycolide) (MPEG-PLGA) microsphere loaded medicine non-prednisone, and the purposes of long-acting insect expelling and safety improvement are realized by controlling the medicine loading, particle size and release behavior of the microsphere.

Specifically, the technical scheme of the invention is as follows:

in a first aspect of the invention, there is provided a non-prednisone drop comprising: a non-splash Luo Niwei ball and a vehicle;

the non-prednisone Luo Niwei spheres comprise non-prednisone and methoxypolyethylene glycol-poly (D, L-lactide-co-glycolide) (MPEG-PLGA);

the solvent comprises a solvent and an antioxidant.

In a specific embodiment, the weight average molecular weight of the MPEG is 1000 Da-2000 Da; the PLGA is 75:25 PLGA, and the weight average molecular weight is 10000 Da-80000 Da, preferably 30000 Da-50000 Da.

In a specific embodiment, the ratio of non-prednisone to MPEG-PLGA in the non-prednisone Luo Niwei spheres is 1 by weight: 3-5.

In a specific embodiment, the particle size of the non-prednisone Luo Niwei spheres is 1-50 μm, and the polydispersity index PDI is 0.05-0.4; preferably, the particle size of the non-splash Luo Niwei spheres is 5-10 mu m, and the polydispersity index PDI is 0.1-0.35.

In a specific embodiment, the dosage ratio of the non-prednisone to the solvent is 50-300 mg:1mL, e.g. 100 mg:1mL,150 mg:1mL,200 mg:1mL; the dosage of the antioxidant is 0.05-0.2% of that of the solvent, and w/v.

Preferably, the solvent is ethanol.

In a specific embodiment, the antioxidant is selected from one or more of Butylhydroxytoluene (BHT), methyl benzoate, propyl benzoate, tert-butylhydroquinone (TBHQ);

preferably, the antioxidant is Butyl Hydroxy Toluene (BHT).

In the non-prednisone drops of the invention, the non-prednisone Luo Niwei balls and the solvent are stored separately and are uniformly mixed to prepare the drops before use in consideration of the stability and drug release properties of the microspheres.

In a second aspect of the present invention, there is provided a method of preparing the non-prednisone Luo Niwei ball, comprising the steps of:

(1) Dissolving non-prednisone and methoxypolyethylene glycol-poly (D, L-lactide-co-glycolide) (MPEG-PLGA) in acetone to obtain oil phase A; dissolving polyvinyl alcohol (PVA) in water to obtain a water phase B, injecting an oil phase A into the water phase B under high-speed shearing, and mixing and emulsifying the two phases to form O/W emulsion;

(2) Drying the O/W emulsion at low temperature under reduced pressure, removing acetone, solidifying the microspheres, and filtering to remove water in the system after solidification to obtain solidified microspheres;

(3) Dispersing the solidified microspheres in polyvinylpyrrolidone (PVP) aqueous solution, and freeze-drying to obtain the non-prednisolone Luo Niwei spheres.

In a specific embodiment, the weight average molecular weight of the PEG in step (1) is 1000 Da-2000 Da; the PLGA is 75:25 PLGA, and the weight average molecular weight is 10000 Da-80000 Da, preferably 30000 Da-50000 Da.

In a specific embodiment, the dosage ratio of the non-prednisone to the MPEG-PLGA is 1:3-5 by weight. The ratio range can obtain microspheres with better drug loading and particle size, and above or below the range, the solution is too viscous to form emulsion, or the drug loading of the microspheres is low or the particle size of the microspheres is uneven or too large.

In a specific embodiment, in the step (1), the mass of the non-prednisone is 10-25% of the mass of the acetone, preferably 10-15%.

In a specific embodiment, in step (1), the polyvinyl alcohol (PVA) is a pharmaceutically acceptable polyvinyl alcohol having an alcoholysis degree of 88% and a weight average molecular weight in the range of 30000 Da~100000 Da, preferably 30000 Da~50000 Da. The mass of the polyvinyl alcohol (PVA) accounts for 0.05-2% of the mass of water, and is preferably 0.08-1.2%. PVA is used as an emulsifier, emulsion liquid drops can be effectively dispersed in the preparation process of O/W emulsion in the concentration range, and the effect of stabilizing the liquid drops is achieved. The inventors have found that the PVA molecular weight and concentration have an effect on the formation and stability of the emulsion, and that the above-mentioned ranges are preferable.

In a specific embodiment, in the step (1), the volume ratio of the oil phase a to the water phase B is 1:1-5, preferably 1:2-3. Above this ratio range, particle size is too small, drug loading is low, and below this ratio range, particle size is too large and the distribution is uneven, thereby affecting drug release.

In a specific embodiment, in the step (1), the rotation speed of the high-speed shearing is 5000-20000 rpm, preferably 10000-15000 rpm.

In a specific embodiment, in step (2), the conditions of drying under reduced pressure at low temperature are: the temperature is 0-25 ℃, and the vacuum degree is 0-0.10 MPa.

In a specific embodiment, in step (3), the polyvinylpyrrolidone (PVP) is selected from model PVP K25 or PVP K30, preferably PVP K30; the mass percentage of polyvinylpyrrolidone (PVP) in the PVP aqueous solution is 1-5%, preferably 2-4%. Polyvinylpyrrolidone (PVP) is used as a freeze-drying protective agent, and the microspheres are dispersed in the freeze-drying protective agent to protect the microsphere structure and support the microspheres. Therefore, the non-prednisone Luo Niwei ball prepared by the preparation method also comprises a freeze-drying protective agent.

In a specific embodiment, the particle size of the non-splash Luo Niwei ball prepared by the preparation method of the non-splash Luo Niwei ball is 1-50 μm, and the polydispersity index PDI is 0.05-0.4; preferably, the particle size is 5-20 μm and the polydispersity index PDI is 0.1-0.35. The microspheres with the particle size range are favorable for entering sebaceous glands and have a long-time slow release effect.

In a third aspect of the present invention, there is provided a method for preparing the non-prednisolone drops according to the first aspect, comprising the steps of:

(1) Dissolving an antioxidant in a solvent to obtain a solvent;

(2) Adding the solvent into the non-prednisone Luo Niwei ball prepared by the preparation method of the non-prednisone Luo Niwei ball in the second aspect of the invention, and uniformly mixing to obtain the non-prednisone drop.

In a specific embodiment, the antioxidant in step (1) is selected from one or more of Butylhydroxytoluene (BHT), methyl benzoate, propyl benzoate, tert-butylhydroquinone (TBHQ).

In a specific embodiment, the ratio of non-prednisone to solvent is: 50-300 mg 1mL, such as 100 mg:1mL,150 mg:1mL,200 mg:1mL; the dosage of the antioxidant is 0.05-0.2% of that of the solvent, and w/v.

In a fourth aspect of the present invention, there is provided a combination comprising: component A: non-splash Luo Niwei balls prepared by the preparation method of the non-splash Luo Niwei balls in the second aspect, and component B: a solvent; the two are stored independently, the storage mode is not limited, and the two can be respectively packaged in penicillin bottles or special plastic bottles convenient for drug administration.

In a fifth aspect of the invention, there is provided the use of a non-prednisone drop as described above or a combination as described above in the manufacture of a medicament for the prophylaxis and/or treatment of a mammalian parasite.

In a specific embodiment, the parasite comprises a flea, a tick.

In a specific embodiment, the mammal comprises a feline, canine, porcine, bovine, equine, donkey, or the like; preferably, the mammal is a feline or canine.

The present invention also provides a method for preventing and/or repelling a mammalian parasite using the non-prednisone drops or the combination drug described above;

specifically, the method comprises the following steps: the non-prednisone drops are used or the components A and B in the combined medicine are uniformly mixed to obtain the drops, then the hairs between the shoulder blades of the animal are separated, and the drops are dripped on the skin along the spine until the absorption is complete.

The invention has the following beneficial effects:

the non-prednisone drop has long-acting drug release property of nearly 2 months, and can greatly prolong the insect expelling period.

The prescription of the drops only contains active ingredients, MPEG-PLGA, solvent ethanol and antioxidant, has simple ingredients, and reduces the irritation of the product to skin; and due to the entrapment and slow drug release characteristics of the microspheres, the drug quantity of the administered animals entering the circulatory system is obviously reduced, the toxic and side effects caused by the drugs are further reduced, and the safety is obviously better than that of the existing products on the market.

Detailed Description

It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.

Based on the problems of the existing insect repellent products, the invention provides a non-prednisone drop and a preparation method and application thereof, and considering that the preparation needs local skin administration and needs to be endeavored to avoid transdermal absorption into a circulatory system, the medicine should be mainly stored in sebaceous glands and released along with grease secreted by the sebaceous glands for a long time. The inventor finds that when a plurality of high polymer materials are used as carrier materials, such as polylactic acid (PLA) or PLGA alone and the like, medicines cannot be released for a long time through the drug loading rate, the encapsulation rate and the animal body long-acting insect repellent experimental study of the preparation; and PLGA is used as a main material to modify PEG with lower molecular weight, so that the drug slow release time can be prolonged. However, too high a molecular weight of PEG modification results in too large a microsphere size, and low drug loading and encapsulation rates, which are not suitable for use in the drug delivery system of the present invention. When the inventor further researches the drug release behaviors of PLGA of different types, the release effects of 95:5 type PLGA and 50:50 type PLGA are not good, and only 75:25 type PLGA can perform ideal release on non-prednisone, so that the carrier material using PEG-PLGA as microspheres is determined. On the basis, the invention controls the particle size and drug loading of the microsphere and further controls the release behavior of the microsphere by perfecting the parameters of the drug auxiliary ratio, the dosage of the emulsifier and the like in the preparation process of the non-prednisone Luo Niwei microsphere. When the microsphere is used in the preparation of drops, verification of the long-acting insect repellent effect of approximately 2 months is obtained in animal experiments, and simultaneously the toxic and side effects are effectively reduced.

The following examples are intended to illustrate embodiments of the invention and are not to be construed as limiting the invention in any way, and it will be understood by those skilled in the art that various modifications may be made without departing from the spirit and scope of the invention.

The MPEG-PLGA structure of each number used in the invention is:

the x, y, n are not limited herein, and this structure is only used to describe the connection of PLGA and MPEG. The specific implementation and the invention of the invention are described for PLGA and MPEG types or molecular weights, and the structures thereof are known in the art.

The MPEG-PLGA materials used in the present invention are all tailored to Chongqing, pharmaceutical technology Co., ltd. All other reagents are commercially available in conventional fashion, and the equipment and testing methods used are conventional in the art, unless otherwise specified.

Example 1

Preparation of non-prednisolone drop-1

(1) 5 g non-prednisone and 25 g of MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) were dissolved in 60 mL acetone to give oil phase A; dissolving 120 mg medicinal PVA (weight average molecular weight of 30000 Da) with alcoholysis degree of 88% in 120 g water to obtain water phase B, injecting the oil phase A into the water phase B under high-speed shearing at 10000 rpm, mixing the two phases, and emulsifying to form O/W emulsion;

(2) Drying the O/W emulsion under reduced pressure at 5 ℃ and minus 0.10 MPa, removing acetone, solidifying the microspheres, and filtering to remove water in the system after solidification to obtain solidified microspheres;

(3) Dispersing the solidified microspheres in 10 mL of 1% polyvinylpyrrolidone PVP K30 aqueous solution, and freeze-drying to obtain non-prednisolone Luo Niwei spheres a;

(4) Dissolving 0.5 mg antioxidant BHT in 1mL of ethanol to obtain a solvent of which the dosage of the antioxidant BHT is 0.05% w/v of that of the solvent ethanol;

(5) And taking a proper amount of the non-prednisone Luo Niwei ball a (containing the non-prednisone 150 mg), adding the solvent into the non-prednisone Luo Niwei ball, and uniformly mixing to obtain the non-prednisone drop-1.

Example 2

Preparation of non-prednisolone drop-2

(1) 5 g non-prednisone and 15 g of MPEG (1500 Da) -PLGA (75:25 type, 50000 Da) were dissolved in 60 mL acetone to give oil phase A; dissolving 120 mg medicinal PVA (weight average molecular weight of 30000 Da) with alcoholysis degree of 88% in 120 g water to obtain water phase B, injecting oil phase A into water phase B under high-speed shearing of 12000 rpm, mixing the two phases, and emulsifying to form O/W emulsion;

(2) Drying the O/W emulsion under reduced pressure at 5 ℃ and minus 0.10 MPa, removing acetone, solidifying the microspheres, and filtering to remove water in the system after solidification to obtain solidified microspheres;

(3) Dispersing the solidified microspheres in 10 mL of 3% polyvinylpyrrolidone PVP K30 aqueous solution, and freeze-drying to obtain non-prednisolone Luo Niwei spheres b;

(4) Dissolving 2 mg antioxidant BHT in 1mL of ethanol to obtain a solvent of which the dosage of the antioxidant BHT is 0.2% w/v of that of the solvent ethanol;

(5) And (3) taking a proper amount of non-prednisone Luo Niwei balls b (containing non-prednisone 150 mg) prepared in the step (3), adding the solvent obtained in the step (4) into the non-prednisone Luo Niwei balls, and uniformly mixing to obtain the non-prednisone drop-2.

Comparative example 1

Preparation of non-prednisone Luo Niwei sphere c:

the difference from example 1 is that: in step (1), MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) was replaced with MPEG (5000 Da) -PLGA (75:25 type, 30000 Da), and in step (3), non-splash Luo Niwei pellets c were produced in the same manner as in example 1. Comparative example 1 only produced non-prednisone Luo Niwei spheres, and no drops were produced.

Comparative example 2

Preparation of non-prednisolone drops-3:

the difference from example 1 is that: in step (1), MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) was replaced with MPEG (1000 Da) -PLGA (50:50 type, 30000 Da), and in step (3), a non-splash Luo Niwei ball d was produced in the same manner as in example 1; and finally obtaining the non-prednisolone drop-3.

Comparative example 3

Preparation of non-prednisolone drops-4:

the difference from example 1 is that: in step (1), MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) was replaced with MPEG (1000 Da) -PLGA (95:5 type, 30000 Da), and in step (3), a non-splash Luo Niwei ball e was produced in the same manner as in example 1; and finally obtaining the non-prednisolone drop-4.

Comparative example 4

Preparation of non-prednisolone drops-5:

the difference from example 1 is that: in step (1), MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) was replaced with PLGA (75:25 type, 30000 Da), and in the same manner as in example 1, a non-splash Luo Niwei ball f was produced in step (3); and finally obtaining the non-prednisolone drop-5.

Comparative example 5

Preparation of non-prednisolone drops-6:

the difference from example 1 is that: in step (1), MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) was replaced with PLA (30000 Da), and in the same manner as in example 1, a non-splash Luo Niwei ball g was produced in step (3); and finally obtaining the non-prednisolone drop-6.

Comparative example 6

Preparation of non-prednisone Luo Niwei sphere h:

the difference from example 1 is that: in the step (1), the use level of MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) is changed from 20 g to 10 g, and in the other steps, the non-splash Luo Niwei ball h is prepared in the step (3) in the same manner as in the example 1; comparative example 6 only a non-prednisone Luo Niwei ball was prepared, and no drops were prepared.

Comparative example 7

Preparation of non-prednisone MPEG-PLGA microspheres:

the difference from example 1 is that: in the step (1), the use level of MPEG (1000 Da) -PLGA (75:25 type, 30000 Da) is changed from 20 g to 30 g, and the system viscosity is found to be too high to form emulsion in the test.

Comparative example 8

Preparation of non-prednisone MPEG-PLGA microspheres:

the difference from example 1 is that: in the step (1), the PVA dosage is changed from 120 mg to 360 mg, and the PVA dosage is found to cause easy foaming in emulsion preparation and can not prepare uniform emulsion in the test.

Comparative example 9

Preparation of non-prednisone MPEG-PLGA microspheres:

the difference from example 1 is that: in the step (1), the PVA dosage is changed from 120 mg to 48 mg, and experiments show that the emulsion is unstable after the preparation is finished and is easy to break.

Test example 1

Physical and chemical property study of non-prednisone Luo Niwei ball

The particle diameters of the non-prednisone Luo Niwei spheres prepared in the above examples and comparative examples were analyzed by a malvern 3000 laser particle sizer, and the drug loading and encapsulation efficiency were measured and calculated by HPLC, and the results are shown in table 1.

TABLE 1

Microsphere(s)	a	b	c	d	e	f	g	h
									Drug loading (%)	12.5	10.3	6.5	8.4	13.1	12.8	11.6	5.2
Encapsulation efficiency (%)	83.2	78.6	40.2	70.3	86.7	84.6	80.1	36.2
									Particle size (mum)	6.2	5.1	31.2	10.9	7.2	6.7	8.7	4.2
PDI	0.213	0.221	0.321	0.287	0.234	0.221	0.265	0.201

As can be seen from the data, the non-prednisolone MPEG-PLGA microspheres prepared in the examples 1-2 have higher drug loading rate and encapsulation rate, and have important significance for encapsulating effective dose of non-prednisolone in the preparation; and the lower particle size makes the microspheres easier to enter pores for storage in sebaceous glands. The microspheres prepared in comparative example 1 using higher molecular weight MPEG have larger particle size and significantly reduced drug loading than the microspheres of the examples, and are not beneficial to skin administration. In addition, the microspheres prepared in comparative examples 2-5 also have acceptable drug loading and particle size, while the drug loading of comparative example 6 is too low to meet the formulation requirements due to too low amount of MPEG-PLGA. Comparative examples 7-9 failed to form an emulsion and failed to complete the preparation of non-prednisole drops.

Test example 2

Animal test

In the embodiment, animal experiments are utilized to research the insect repellent effect of the non-prednisone Luo Niwei balls after being prepared into drops.

Test animals: the dogs infected with ticks were confirmed by clinical examination and laboratory diagnosis, and had not received insect repellent treatment for nearly 6 months; the number of infected ticks of the test dogs before the test is more than or equal to 1, the variety and the sex are unlimited, and the age is 8 weeks or more.

Preparation: non-prednisone drop-1 prepared in example 1, non-prednisone drop-2 prepared in example 2, and non-prednisone drops 3-6 prepared in comparative examples 2-5 (containing 150 mg non-prednisone, 1 mL); control drug: fulaien drops (specification: 100 mg non-prednisone+90 mg methoprene, 0.67 mL).

The test method comprises the following steps:

the test animals were divided into 7 groups of 6 animals each, and the above formulation and the control drug were administered separately. The administration method comprises the following steps: the hair at the scapula of the test dog is allocated, the medicine is applied to the skin in a dripping way along the spinal column, and the medicine is completely absorbed for 1 time, and the test dog does not have a bath after the medicine is applied until the test is finished.

The administration doses are respectively as follows: test dogs weighing 10 kg or less used 1mL non-prednisolone drops 1-6 or 0.67 mL flean drops; 2 mL non-prednisolone drops 1-6 or 1.34 mL Fulain drops 1.34 mL are used for test dogs weighing 10-20 kg; 3mL of non-prednisolone drops 1-6 or 2.68 mL Fulain drops are used for test dogs with the weight of 20-40 kg; for test dogs with a body weight greater than 40 kg, 3 mL+n×1mL non-prednisolone drops 1-6 or 2.68 mL+n×0.67 mL of Fulain drops were used for every n increments of 10 kg.

The test period was 60 days, the skin states were observed on the day of administration (0 d), 3 d, 7 d, 14 d, 21 d, 28 d, 35 d, 42 d, 49 d, 56 d, 60 d after administration, respectively, and the number of body surface ticks was collected and recorded, and the results are shown in table 2.

Table 2.

Drops (drops)

0d

3d

7d

14d

21d

28d

35d

42d

49d

56d

60d

1

3.17±0.75

0.17±0.41

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.16±0.41

2

3.17±1.60

0.33±0.52

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.16±0.41

0.33±0.82

3

2.83±1.47

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.50±0.84

0.83±0.98

1.83±1.17

2.33±0.82

3.00±0.63

4

3.17±2.04

1.83±1.17

1.00±1.09

0.00±0.00

0.00±0.00

0.33±0.52

0.83±0.75

1.33±0.82

1.67±1.03

2.17±1.17

2.67±1.37

5

3.17±1.17

1.50±1.05

0.67±0.82

0.00±0.00

0.00±0.00

1.00±0.63

1.33±1.03

1.67±1.37

2.00±1.41

2.50±1.64

3.00±1.41

6

3.00±1.10

1.33±0.52

0.83±0.75

0.00±0.00

0.00±0.00

1.67±0.98

1.50±1.04

1.67±1.03

2.17±0.75

2.67±1.37

3.33±1.21

Control

2.83±1.26

0.00±0.00

0.00±0.00

0.00±0.00

0.00±0.00

0.17±041

1.17±0.75

1.83±0.41

2.33±1.03

3.00±1.42

3.17±1.17

As can be seen from Table 2, the control drug, fulain drops, was effective the third day after use, and reinfection with ticks was observed on day 28. Whereas the non-prednisolone drops-1 prepared in example 1 and the non-prednisolone drops-2 prepared in example 2 of the present invention have slightly slower onset of action, a decrease in the number of tick infections is observed on the third day, which may be related to the need for slow release of the drug from the microspheres, but from the viewpoint of long-term anthelmintic effect, the non-prednisolone drops-1, non-prednisolone drops-2 re-appear tick infections after day 56 or day 60 after administration, indicating that the microspheres have a long-term slow release effect. Non-prednisone drop-3 prepared with 50:50 type PLGA as comparative example 2 (non-prednisone drop-3) had a shorter onset of action and had an overall insect repellent effect on day 3 after administration, which may be associated with faster administration of microspheres prepared with 50:50 type PLGA, but had poor long-term administration. Use 95: the delay in the insect repellent effect was also observed for non-prednisone drops prepared from type 5 PLGA (non-prednisone drop-4), simple PLGA (non-prednisone drop-5) or simple PLA (non-prednisone drop-6), but worse, the re-infection of ticks occurred on day 28, possibly due to the inability of the corresponding microspheres to remain in sebaceous glands for a long period of time, or the low drug accumulation release rate of the microspheres.

In addition, on day 3 of administration of the control drug flene drop, 1 occurrence of skin rash was observed, which resolved within one week after that. The non-prednisone drops prepared in examples 1 and 2 and comparative examples 2-5 of the present invention did not show obvious skin rash and itching symptoms in the whole course of the test, indicating that the safety of the drops of the present invention is higher.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A non-prednisone drop, comprising: a non-splash Luo Niwei ball and a vehicle;

the non-prednisone Luo Niwei spheres comprise non-prednisone and MPEG-PLGA;

the solvent comprises a solvent and an antioxidant.

2. The non-prednisone drop of claim 1, wherein said MPEG has a weight average molecular weight of 1000 Da to 2000Da; the PLGA is 75:25 PLGA, and the weight average molecular weight is 10000 Da-80000 Da.

3. The non-prednisone drop of claim 1, wherein the ratio of non-prednisone to MPEG-PLGA in said non-prednisone Luo Niwei sphere is 1 by weight: 3-5.

4. The non-prednisone drop of claim 1, wherein said non-prednisone Luo Niwei spheres have a particle size of 1-50 μm and a polydispersity index PDI of 0.05-0.4.

5. The non-prednisone drop of claim 1, wherein the dosage ratio of non-prednisone to solvent is 50-300 mg: 1. 1mL, wherein the dosage of the antioxidant is 0.05-0.2% of the solvent, and w/v.

6. A non-prednisone drop according to claim 1, wherein said antioxidant is selected from one or more of butylhydroxytoluene, methyl paraben, propyl paraben, tert-butylhydroquinone.

7. The non-prednisone drop of claim 1, wherein said non-prednisone Luo Niwei pellet is prepared by a process comprising the steps of:

(1) Dissolving non-prednisone and MPEG-PLGA in acetone to obtain an oil phase A; PVA is dissolved in water to obtain a water phase B, the oil phase A is injected into the water phase B under high-speed shearing, and two phases are mixed and emulsified to form O/W emulsion;

(2) Drying the O/W emulsion at the temperature of 0-25 ℃ under reduced pressure, removing acetone, solidifying the microspheres, and filtering to remove water in the system after solidification to obtain solidified microspheres;

(3) Dispersing the solidified microspheres in PVP aqueous solution, and freeze-drying to obtain the non-prednisolone Luo Niwei spheres.

8. A method of preparing a non-prednisone drop as claimed in any one of claims 1 to 7, comprising the steps of:

(1) Dissolving an antioxidant in ethanol to obtain a solvent;

(2) Adding the solvent into the non-prednisone Luo Niwei spheres, and uniformly mixing to obtain the non-prednisone drops.

9. A combination medicament, comprising: component A: non-prednisone Luo Niwei spheres according to claim 1, component B: the vehicle of claim 1; both are stored independently.

10. Use of a non-prednisone drop according to any one of claims 1 to 7 and/or a combination according to claim 9 in the manufacture of a medicament for the prevention and/or the expelling of parasites in mammals.