Compound fipronil praziquantel gel as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to compound fipronil praziquantel gel as well as a preparation method and application thereof. The compound fipronil praziquantel gel is prepared from fipronil, praziquantel, acetamido abamectin, metopropene, poloxamer 407, a penetration enhancer, hydroxypropyl methyl cellulose, a preservative, absolute ethyl alcohol and water. The fipronil and praziquantel gel is prepared from the following components in percentage by mass and volume: 6.225% of fipronil, 6.225% of praziquantel, 0.3% of acetamido abamectin, 7.5% of metopropene, 10%-13% of poloxamer 407, 6%-15% of a penetration enhancer, 0.05%-0.1% of potassium sorbate, 0-1.67% of hydroxypropyl methyl cellulose HPMC, 28%-30% of absolute ethyl alcohol and the balance of water. The compound fipronil praziquantel gel prepared by the invention not only has a good transdermal effect, but also can prolong the drug release time, and has an obvious slow release effect.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a compound fepronil praziquantel gel and a preparation method and application thereof. Background Art

[0002] The compound fepronil praziquantel is composed of fepronil, methoprene, avermectin, and praziquantel. It is used to prevent and treat flea and tick infections in cats, treat gastrointestinal nematode and tapeworm infections, and can be used as an auxiliary treatment for allergic dermatitis caused by fleas.

[0003] The pure product of fenprodil is a white crystalline powder, and its chemical name is (RS)-5-amino-1-(2,6-dichloro-4-trifluoromethylphenyl)-4-trifluoromethyl, which is a broad-spectrum insecticide. It binds to the γ-aminobutyric acid (GABA) receptors on the membrane of insect central nervous cells, closes the chloride ion channels of nerve cells, and thus interferes with the normal function of the central nervous system and causes the death of insects.

[0004] Methoxypromide is a colorless or light yellow oily liquid. Its chemical name is (7S, 2E, 4E)-11-methoxy-3,7,11-trimethyldodecadienoic acid isopropyl ester. It is an insect growth regulator (IGR) and a congener of insect juvenile hormone. It has an inhibitory effect on the development of immature insects. This drug has a similar mechanism of action to juvenile hormone and can cause insect development stunting and flea death in the developmental stage.

[0005] The raw material of avermectin is a white crystalline solid at room temperature. It has a strong lipophilic group in its molecular structure and is highly lipophilic. It has a good expelling and killing effect on parasites inside and outside the body, mainly nematodes and arthropods. Its anthelmintic mechanism is to promote the release of γ-aminobutyric acid (GABA) from precontactile neurons, thereby opening GABA-mediated chloride ion channels. Acetaminobamectin also has selectivity and high affinity for glutamate-mediated chloride ion channels located near GABA-mediated points in nerve and muscle cells of invertebrates, thereby interfering with signal transmission between nerves and muscles, causing the parasite to relax and paralyze, leading to the death of the parasite or being excreted from the body.

[0006] Praziquantel is a white or off-white crystalline powder with a broad spectrum of anti-schistosomal and anti-tapeworm effects. It has good activity against adults and larvae of various tapeworms. The exact mechanism of action of praziquantel on tapeworms has not yet been determined, but it may be an interaction with the phospholipids of the worm's capsule, resulting in the outflow of sodium and potassium ions. In vitro, low concentrations of praziquantel appear to damage the tapeworm's sucker function and excite the worm's peristalsis, while higher concentrations of the drug can enhance the contraction of the tapeworm chain (segment chain) (irreversible at extremely high concentrations). In addition, praziquantel can cause focal vacuoles to form in special parts of the tapeworm's capsule, which then causes the worm to lyse.

[0007] At present, compound fepronil praziquantel has only one dosage form: drops. The more mature product on the market is compound fepronil praziquantel drops for external use produced by Boehringer Ingelheim Animal Health (China) Co., Ltd., with the Chinese trade name "Bolaien". In the drops, each 1ml contains 83mg of fepronil, 100mg of methoprene, 4mg of avermectin, and 83mg of praziquantel. The preparation is dripped on the skin for use, which has the characteristics of broad-spectrum deworming in vivo and in vitro and convenient use. However, ordinary transdermal drops have a short residence time on the body surface, and animals will shake, resulting in incomplete drug penetration, affecting the efficacy of fepronil, praziquantel, avermectin, and methoprene. After treatment, transient hair adhesion and mild transient skin reactions (itching, hair loss) may be found at the medication site. In addition, the commercially available Bolaien products are more oily for animal hair, and the feeling of use is relatively poor.

[0008] Therefore, the development of a compound non-prednisolone praziquantel preparation that is free of loss, highly permeable, low in oiliness, safe and mild, and has high efficacy and long-lasting efficacy has important practical application value and is the direction that pharmaceutical technicians are working hard to research. Summary of the invention

[0009] In order to overcome the defects of the prior art, the present invention provides a compound non-pronil praziquantel gel, which has low oiliness, constant temperature and breathability, good sustained release effect, long duration, and is conducive to the anthelmintic effect.

[0010] The present invention is achieved through the following technical solutions:

[0011] A compound fepronil praziquantel gel, comprising fepronil, praziquantel, avermectin, methoprene, poloxamer 407, a penetration enhancer, hydroxypropyl methylcellulose, a preservative, anhydrous ethanol and water; the mass volume percentage of each component in the gel is: 6.225% of fepronil, 6.225% of praziquantel, 0.3% of avermectin, 7.5% of methoprene, 10%-13% of poloxamer 407, 6%-15% of a penetration enhancer, 0.05-0.1% of potassium sorbate, 0-1.67% of hydroxypropyl methylcellulose HPMC, 28-30% of anhydrous ethanol, and the balance of water. The penetration enhancer is one or a combination of two of 1,2-propylene glycol, polyethylene glycol 400 and laurocapram.

[0012] Preferably, the penetration enhancer is a combination of 1,2-propylene glycol and laurocapram.

[0013] Further, the present invention preferably comprises the following compound fepronil praziquantel gel, wherein the mass volume percentage of each component in the compound fepronil praziquantel gel is: fepronil 6.225%, praziquantel 6.225%, avermectin 0.3%, methoprene 7.5%, poloxamer 407 10%-13%, 1,2-propylene glycol 5%-10%, laurocapram 1%-5%, potassium sorbate 0.05-0.1%, anhydrous ethanol 28-30%, and water as the balance.

[0014] Further, the present invention preferably comprises the following gel, wherein the weight volume percentage of each component in the gel is: 6.225% of fepronil, 6.225% of praziquantel, 0.3% of acetaminophen, 7.5% of methoprene, 11%-12% of poloxamer 407, 6%-8% of 1,2-propylene glycol, 3%-5% of laurocapram, 0.05-0.1% of potassium sorbate, 28-30% of anhydrous ethanol, and the balance of water.

[0015] Furthermore, the mass ratio of Poloxamer 407 to the total mass of 1,2-propylene glycol and laurocapram is 0.8-1.2:1, preferably 0.9-1.1:1.

[0016] The mass ratio of laurocapram to 1,2-propylene glycol is 1:1.8-1:2.2, preferably 1:2.0-2.1.

[0017] Furthermore, in the compound nonpronil praziquantel gel of the present invention, the HPMC is HPMC E5 or E50, and the mass volume percentage of the HPMC is less than 0.42%.

[0018] When the HPMC is HPMC E50, its mass volume percentage is preferably: 0.25-0.42%, preferably 0.25%;

[0019] When the HPMC is HPMC E5, its mass volume percentage is preferably: 0.33%-0.42%, preferably 0.33%.

[0020] The preparation process of the gel of the present invention is:

[0021] (1) Accurately weigh poloxamer 407, add poloxamer 407 to distilled water, seal and place in a 4°C refrigerator for 24 hours to obtain an aqueous solution of poloxamer 407;

[0022] (2) adding a preservative, hydroxypropyl methylcellulose and a penetration enhancer to the aqueous solution of poloxamer 407, stirring evenly, sealing and standing to obtain a matrix solution;

[0023] (3) Add fenprodil, praziquantel, avermectin, and methoprene to anhydrous ethanol and dissolve them by ultrasonication. Add the drug solution to the above matrix solution, stir evenly, and ultrasonicate in a low-temperature ultrasonic instrument until the drug is completely dissolved. Seal and let stand.

[0024] (4) Add distilled water to a certain volume and stir evenly to obtain a gel.

[0025] The prepared compound fepronil praziquantel gel is 1.2 ml / tube, and the preparation can be used for animals weighing 2.5 kg to 7.5 kg, and each 1.2 ml contains 74.7 mg of fepronil, 90.0 mg of methoprene, 3.6 mg of acetaminophen, and 74.7 mg of praziquantel; for animals weighing less than 2.5 kg, 0.4 ml of the preparation is used, and each 0.4 ml contains 24.90 mg of fepronil, 30.00 mg of methoprene, 1.20 mg of acetaminophen, and 24.90 mg of praziquantel. The preparation is usually sealed and stored at room temperature (not higher than 25°C).

[0026] The present invention uses gelation experiment and rat transdermal experiment as research methods, and uses gelation temperature, gelation time and transdermal duration as evaluation indicators to examine the formulation of the gel, thereby obtaining a gel with good transdermal penetration-enhancing effect and long sustained-release time.

[0027] The experimental results show that the compound nonpronil praziquantel gel prepared by the present invention not only has a good transdermal effect, but also can prolong the release time of the drug and has an obvious sustained-release effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a graph of the gelation temperatures of Example 3, Example 8, Example 11 and Example 17.

[0029] Figure 2 This is a graph of the gel time of Example 3, Example 8, Example 11 and Example 17.

[0030] Figure 3 This is a comparison chart of the cumulative drug permeation of the drug praziquantel in Example 3 and the commercially available "Bolain".

[0031] Figure 4 This is a comparison chart of the cumulative drug permeation over time of the drug fepronil in Example 3 and the commercially available "Bolaien".

[0032] Figure 5 This is a comparison chart of the cumulative drug permeation of methoprene over time in Example 3 and the commercially available "Bolain".

[0033] Figure 6 This is a comparison chart of the cumulative drug permeation of avermectin over time in Example 3 and the commercially available "Bolain".

[0034] Figure 7 This is a comparison chart of the cumulative drug permeation of the drug praziquantel over time in Example 8, Example 11 and Example 12.

[0035] Figure 8 This is a comparison chart of the cumulative drug permeation of the drug fepronil over time in Example 8, Example 11 and Example 12.

[0036] Fig. 9 This is a comparison chart of the cumulative drug permeation of methoprene over time in Example 8, Example 11 and Example 12.

[0037] Fig.10 This is a comparison chart of the cumulative drug permeation of avermectin over time in Example 8, Example 11 and Example 12.

[0038] Fig.11 This is a comparison chart of the cumulative drug permeation of the drug praziquantel over time in Example 15, Example 16 and Example 17.

[0039] Fig.12 This is a comparison chart of the cumulative drug permeation of the drug fepronil over time in Example 15, Example 16 and Example 17.

[0040] Fig.13 This is a comparison chart of the cumulative drug permeation of methoprene over time in Example 15, Example 16 and Example 17.

[0041] Fig.14 This is a comparison chart of the cumulative drug permeation of avermectin over time in Example 15, Example 16 and Example 17. DETAILED DESCRIPTION

[0042] The present invention is further described below by way of examples, but the present invention is not limited to the following examples. Without departing from the above technical premise of the present invention, corresponding replacements or modifications made according to common technical knowledge and conventional means in the art are all included in the scope of the present invention.

[0043] The preparations prepared in the following examples total 120 ml, each of which has a volume of 1.2 ml (74.7 mg of fepronil, 90 mg of methoprene, 3.6 mg of avermectin, and 74.7 mg of praziquantel), and the drug content contained is consistent with the drug content contained in the commercially available "Bolain" 0.9 ml / tube drops.

[0044] Embodiment 1:

[0045] Poloxamer 407 12.60g 10.5％ Laurocapram 8.28g 6.9％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0046] Preparation method:

[0047] (1) Accurately weigh the prescribed amount of poloxamer 407 and potassium sorbate, add distilled water, seal and place in a 4°C refrigerator for 24 hours to obtain a gel matrix solution;

[0048] (2) accurately weighing the prescribed amount of fepronil, praziquantel, avermectin, and methoprene, adding anhydrous ethanol, dissolving them by low-temperature ultrasonication, and then adding them to the gel matrix prepared in step (1), stirring evenly, and ultrasonicating them in a low-temperature ultrasonic instrument until the drugs are completely dissolved, and then sealing and letting them stand;

[0049] (3) Accurately weigh the prescribed amount of laurocapram and add it to the above solution, stir evenly, add distilled water to 120 ml, stir evenly, and divide into portions (specification: 1.2 ml) to obtain the gel.

[0050] Determination of gelation temperature of gel: Take 1.2 ml at room temperature, when the preparation is in sol state. Place the EP test tube in a water bath, and observe the temperature when the sol turns into gel under different temperature conditions, which is the gelation temperature. Each example is measured three times and the average value is calculated.

[0051] Determination of gelation time of gel: Take 1.2 ml at room temperature, when the preparation is in a sol state. Place the EP test tube in a water bath at 32°C, start timing, and invert the EP test tube from time to time to observe the gelation time of the sol, which is the gelation time of the gel. Each example is measured three times and the average value is calculated.

[0052] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature, but is placed in a water bath, the gelling temperature exceeds 35°C, and then placed at room temperature to restore fluidity.

[0053] Embodiment 2:

[0054] Poloxamer 407 15.20g 12.67％ 1,2-Propanediol 5.18g 4.32％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0055] Preparation method:

[0056] Same as Example 1.

[0057] Determination of gelation temperature and time:

[0058] Same as Example 1.

[0059] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature. It is placed in a water bath and gelled at 33°C, with a gelling time of 69s. It does not recover its fluidity when placed at room temperature.

[0060] Embodiment 3:

[0061]

[0062]

[0063] Preparation method:

[0064] Same as Example 1.

[0065] Determination of gelation temperature and time:

[0066] Same as Example 1.

[0067] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature, but quickly gels at body temperature (32°C), with a gelling time of 50 seconds. It then returns to fluidity when placed at room temperature.

[0068] Embodiment 4:

[0069] Poloxamer 407 14.30g 11.92％ 1,2-Propanediol 5.18g 4.32％ Potassium Sorbate 0.06g 0.05％ Laurocapram 8.28g 6.90％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0070] Preparation method:

[0071] Same as Example 1.

[0072] Determination of gelation temperature and time:

[0073] Same as Example 1.

[0074] This product is homogeneous and does not separate after standing. It is a milky white liquid. It gels at room temperature.

[0075] Embodiment 5:

[0076]

[0077]

[0078] Preparation method:

[0079] Same as Example 1.

[0080] Determination of gelation temperature and time:

[0081] Same as Example 1.

[0082] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel when the water bath temperature exceeds 40°C, and still has fluidity when placed at room temperature.

[0083] Embodiment 6:

[0084] Poloxamer 407 14.30g 11.92％ Laurocapram 12.88g 10.73％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0085] Preparation method:

[0086] Same as Example 1.

[0087] Determination of gelation temperature and time:

[0088] Same as Example 1.

[0089] This product is homogeneous and does not separate after standing. It is a milky white liquid. It gels at room temperature.

[0090] Embodiment 7:

[0091]

[0092]

[0093] Preparation method:

[0094] Same as Example 1.

[0095] Determination of gelation temperature and time:

[0096] Same as Example 1.

[0097] The product is homogeneous and does not separate after standing. It is a milky white liquid. It does not gel at room temperature, but quickly gels at 34°C in a water bath with a gelling time of 50 seconds. It does not recover fluidity when left at room temperature.

[0098] In Examples 1-7, the gelling temperature and gelling time of the gelling agent were investigated under the conditions of changing the dosage of poloxamer 407 and the type and dosage of the penetration enhancer. In Examples 1 and 6, different proportions of laurocapram were used as penetration enhancers, but gelling agents with qualified gelling temperatures could not be obtained. In Examples 2, 4 and 5, different proportions of 1,2-propylene glycol were used as penetration enhancers, but gelling agents with qualified gelling temperatures and fluidity at room temperature could not be obtained. In Example 3, a combination of laurocapram and 1,2-propylene glycol was used as a penetration enhancer, and the mass ratio of laurocapram to 1,2-propylene glycol was about 1:2, and the mass ratio of poloxamer 407 to the total mass of the combination of laurocapram and 1,2-propylene glycol was about 1:1, and a gelling agent with qualified gelling temperature and fluidity could be prepared.

[0099] According to the results of Examples 1-7, the present invention selects the formulation of Example 3 to prepare the compound fepronil praziquantel gel.

[0100] On the basis of Example 3, the amounts of poloxamer 407, 1,2-propylene glycol and laurocapram were changed. The results showed that when the mass ratio of poloxamer 407 to the sum of 1,2-propylene glycol and laurocapram was 1:0.8-1.2, and the mass ratio of laurocapram to 1,2-propylene glycol was 1:1.8-1:2.2, a gel having both a qualified gelling temperature and fluidity at room temperature could be obtained. Outside this range, a gel having both qualified gelling temperature and fluidity could not be obtained.

[0101] Embodiment 8:

[0102] Preliminary tests have shown that the addition of HPMC may increase drug penetration. Therefore, the present invention uses the formulation of Example 3 as the basic formulation, adds HPMC of different types (E50 or E5) and different concentrations to investigate the effects of the type and amount of HPMC on the gelling temperature, gelling time and drug penetration in the gel.

[0103]

[0104]

[0105] Preparation method:

[0106] (1) Accurately weigh the prescribed amount of poloxamer 407, HPMC E50 and potassium sorbate, add distilled water, seal and place in a 4°C refrigerator for 24 hours to obtain a gel matrix solution;

[0107] (2) accurately weighing the prescribed amount of fepronil, praziquantel, avermectin, and methoprene, adding anhydrous ethanol, dissolving them by low-temperature ultrasonication, and then adding them to the gel matrix prepared in step (1), stirring evenly, and ultrasonicating them in a low-temperature ultrasonic instrument until the drugs are completely dissolved, and then sealing and letting them stand;

[0108] (3) Accurately weigh the prescribed amount of 1,2-propylene glycol and laurocapram, add them to the above solution, stir evenly, add distilled water to 120 ml, stir evenly, and divide into portions (specification: 1.2 ml) to obtain the gel.

[0109] Determination of gelation temperature of gel: Take 1.2 ml into EP test tube at room temperature, at which time the preparation is in sol state. Place the EP test tube in a water bath, and observe the temperature when the sol turns into gel under different temperature conditions, which is the gelation temperature. Each example is measured three times and the average value is calculated.

[0110] Determination of gelation time of gel: Take 1.2 ml at room temperature, when the preparation is in a sol state. Place the EP test tube in a water bath at 32°C, start timing, and invert the EP test tube from time to time to observe the gelation time of the sol, which is the gelation time of the gel. Each example is measured three times and the average value is calculated.

[0111] The product is homogeneous and does not separate after standing. It is a milky white liquid. It does not gel at room temperature. It is placed in a water bath and quickly gels at body temperature (32°C). The gelation time is 92s. It is then placed at room temperature to restore fluidity.

[0112] Embodiment 9:

[0113]

[0114]

[0115] Preparation method:

[0116] Same as Example 8.

[0117] Determination of gelation temperature and time:

[0118] Same as Example 8.

[0119] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature. It is placed in a water bath with a gelling temperature of 33°C. It is then placed at room temperature to restore fluidity.

[0120] Embodiment 10:

[0121] Poloxamer 407 14.30g 11.92％ HPMC E50 2.00g 1.67％ 1,2-Propanediol 9.32g 7.77％ Laurocapram 4.60g 3.83％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0122] Preparation method:

[0123] Same as Example 8.

[0124] Determination of gelation temperature and time:

[0125] Same as Example 8.

[0126] The product is homogeneous and does not separate after standing. It is a milky white liquid. It does not gel at room temperature. It is placed in a water bath and quickly gels at body temperature (32°C). The gelation time is 27s. It does not recover its fluidity when placed at room temperature.

[0127] The results of Examples 8-10 show that the gelling temperature and gelling time of the preparations are different when the concentration of HPMC E50 is changed. When the percentage of HPMC E50 is 0.42%, the gelling temperature of the preparation increases. When the percentage of HPMC E50 is 1.67%, although the gelling temperature of the preparation is qualified, the fluidity cannot be restored at room temperature. Therefore, when HPMC E50 is added, the percentage of HPMC E50 should be less than 0.42%, preferably 0.25%, at which time the drug matrix is ​​in the best state.

[0128] Embodiment 11:

[0129] Poloxamer 407 14.30g 11.92％ HPMC E5 0.40g 0.33％ 1,2-Propanediol 9.32g 7.77％ Laurocapram 4.60g 3.83％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0130] Preparation method:

[0131] Same as Example 8.

[0132] Determination of gelation temperature and time:

[0133] Same as Example 8.

[0134] The product is homogeneous and does not separate after standing. It is a milky white liquid. It does not gel at room temperature. It is placed in a water bath and quickly gels at body temperature (32°C). The gelation time is 36s. It is then placed at room temperature to restore fluidity.

[0135] Embodiment 12:

[0136]

[0137]

[0138] Preparation method:

[0139] Same as Example 8.

[0140] Determination of gelation temperature and time:

[0141] Same as Example 8.

[0142] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature. It is placed in a water bath with a gelling temperature of 34°C. It is then placed at room temperature to restore fluidity.

[0143] Embodiment 13:

[0144] Poloxamer 407 14.30g 11.92％ HPMC E5 1.00g 0.84％ 1,2-Propanediol 9.32g 7.77％ Laurocapram 4.60g 3.83％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0145] Preparation method:

[0146] Same as Example 8.

[0147] Determination of gelation temperature and time:

[0148] Same as Example 8.

[0149] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature, but is placed in a water bath, the gelling temperature exceeds 35°C, and then placed at room temperature to restore fluidity.

[0150] Embodiment 14:

[0151]

[0152]

[0153] Preparation method:

[0154] Same as Example 8.

[0155] Determination of gelation temperature and time:

[0156] Same as Example 8.

[0157] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature, but is placed in a water bath, the gelling temperature exceeds 35°C, and then placed at room temperature to restore fluidity.

[0158] The results of Examples 11-14 show that the gelling temperature and gelling time of the preparations are different when the concentration of HPMC E5 is changed. When the percentage of HPMC E5 is 0.84% ​​and 1.67%, the gelling temperature of the preparations exceeds 35°C, which does not meet the gelling temperature requirement. Therefore, the percentage of HPMC E5 should be less than 0.42%, preferably 0.33%, at which time, the drug matrix is ​​in the best state.

[0159] As shown in the results of Examples 8-14, when other ingredients remain unchanged, adding different amounts of HPMC E5 or E50 will affect the gelling temperature and gelling time of the gelling agent.

[0160] Embodiment 15:

[0161] The formulation of the gel was determined by the above method. When the dosages of poloxamer 407 and HPMC E5 were fixed, the new penetration enhancer crotamiton was used as the penetration enhancer in the formulation to investigate its effect on the gelation temperature and gelation time of the gel.

[0162]

[0163]

[0164] (1) Accurately weigh the prescribed amount of poloxamer 407, HPMC E5 and potassium sorbate, add distilled water, seal and place in a 4°C refrigerator for 24 hours to obtain a gel matrix solution;

[0165] (2) accurately weighing the prescribed amount of fepronil, praziquantel, avermectin, and methoprene, adding anhydrous ethanol, dissolving them by low-temperature ultrasonication, and then adding them to the gel matrix prepared in step (1), stirring evenly, and ultrasonicating them in a low-temperature ultrasonic instrument until the drugs are completely dissolved, and then sealing and letting them stand;

[0166] (3) Accurately weigh the prescribed amount of crotamiton and add it to the above solution, stir evenly, add distilled water to 120 ml, stir evenly, and divide into portions (specification: 1.2 ml) to obtain the gel.

[0167] Determination of gelation temperature of gel: Take 1.2 ml into EP test tube at room temperature, at which time the preparation is in sol state. Place the EP test tube in a water bath, and observe the temperature when the sol turns into gel under different temperature conditions, which is the gelation temperature. Each example is measured three times and the average value is calculated.

[0168] Determination of gelation time of gel: Take 1.2 ml at room temperature, when the preparation is in a sol state. Place the EP test tube in a water bath at 32°C, start timing, and invert the EP test tube from time to time to observe the gelation time of the sol, which is the gelation time of the gel. Each example is measured three times and the average value is calculated.

[0169] This product is homogeneous and does not separate after standing. It is a milky white liquid. It gels at room temperature.

[0170] Embodiment 16:

[0171] Poloxamer 407 14.30g 11.92％ HPMC E5 0.40g 0.33％ Cromiton 13.92g 11.6％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0172] Preparation method:

[0173] Same as Example 15.

[0174] Determination of gelation temperature and time:

[0175] Same as Example 15.

[0176] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature, but is placed in a water bath, the gelling temperature exceeds 35°C, and then placed at room temperature to restore fluidity.

[0177] The results of Example 15 and Example 16 show that, when the other ingredients and dosages in the formula are fixed and the penetration enhancer is HPMC E5, replacing the penetration enhancer with crotamiton, a gel with a qualified gelling temperature and fluidity cannot be obtained.

[0178] Embodiment 17:

[0179] Poloxamer 407 14.30g 11.92％ HPMC E5 0.40g 0.33％ 1,2-Propanediol 9.32g 7.77％ Laurocapram 4.60g 3.83％ Cromiton 0.45g 0.38％ Potassium Sorbate 0.06g 0.05％ Fipronil 7.47g 6.225％ Methoxyprene 9.00g 7.5％ Avermectin 0.36g 0.3％ Praziquantel 7.47g 6.225％ Anhydrous ethanol 36.00ml 30％ Distilled water Add to 120ml margin

[0180] Preparation method:

[0181] Same as Example 15.

[0182] Determination of gelation temperature and time:

[0183] Same as Example 15.

[0184] The product is uniform in state, does not separate after standing, and is a milky white liquid. It does not gel at room temperature. It is placed in a water bath with a gelling temperature of 33°C and a gelling time of 120s. It is then placed at room temperature to restore fluidity.

[0185] In Example 17, in which crotamiton is added to Example 11, the gelling temperature is slightly increased, but it basically meets the qualified standards of the gel.

[0186] In summary, different penetration enhancer types and dosages have different impacts on the gelling temperature of gel. When the penetration enhancer is 1, under the condition of the combination of 2-propylene glycol and laurocapram, a gelling temperature qualified can be obtained. And under the condition of the combination of 1,2-propylene glycol and laurocapram, crotamiton is added, which has little impact on its gelling temperature, but the gelling time is prolonged.

[0187] Experimental Example 1 Gel Transdermal Test

[0188] In the following experiment, the present invention selected preparations that gel at body temperature (32°C), namely, Examples 3, 8 and 11, as well as Example 12 with a slightly elevated gelling temperature but better fluidity at room temperature, and Example 17 with crotamiton added to Example 11 for subsequent rat transdermal experiments.

[0189] Rat transdermal test

[0190] This experiment used a vertical diffusion cell. The rat skin was fixed between the drug delivery cell and the receiving cell. The stratum corneum layer faced the delivery chamber and the dermis layer faced the receiving chamber. The effective diffusion area was 1.54 cm 2 . Take the gels of Examples 3, 8, 11, 12 and 17 and the commercially available "Bolain" drops respectively and add them to the dosing pool, evenly apply them on the rat skin, and take 4 ml of the prepared receiving solution (30% ethanol and 70% saline) and add it to the receiving pool, remove bubbles, and make the rat skin contact with the liquid surface. Continue stirring at 32°C and 500 (r / min), extract 4 ml of the receiving solution 5min, 10min, 15min, 20min, 25min, 30min, 1h, 2h, 6h, 12h, 24h, 36h, 48h after administration and supplement with fresh receiving solution of the same volume and temperature, filter the extracted receiving solution with a 0.45μm filter membrane to obtain the filtrate, measure the ultraviolet absorbance, and calculate the cumulative penetration amount Q.

[0191]

[0192] ρn is the mass concentration of the nth sampling point (μg·mL －1 ), V is the sampling volume, and A is the penetration area.

[0193] The results of three parallel transdermal tests on rats showed that:

[0194] The cumulative transdermal release of praziquantel in the commercially available "Bolain" test product was 11.34 mg, and the drug sustained-release time was estimated to be 109 hours. The cumulative transdermal release of pyroptidine was 0.81 mg, and the drug sustained-release time was estimated to be 23 hours. The cumulative transdermal release of methoprene was 6.01 mg, and the drug sustained-release time was estimated to be 784 hours. The cumulative transdermal release of avermectin was 0.59 mg, and the drug sustained-release time was estimated to be 312 hours.

[0195] The cumulative transdermal release of the praziquantel drug in Example 3 is 10.08 mg, and the drug sustained release time is calculated to be 367 h. The cumulative transdermal release of the pyroptidine drug is 0.71 mg, and the drug sustained release time is calculated to be 5364 h. The cumulative transdermal release of the methoprene drug is 5.37 mg, and the drug sustained release time is calculated to be 823 h. The cumulative transdermal release of the avermectin drug is 0.59 mg, and the drug sustained release time is calculated to be 290 h.

[0196] The cumulative transdermal release of the praziquantel drug in Example 8 is 15.22 mg, and the drug sustained release time is calculated to be 225 h. The cumulative transdermal release of the pyroptidine drug is 0.63 mg, and the drug sustained release time is calculated to be 6429 h. The cumulative transdermal release of the methoprene drug is 11.19 mg, and the drug sustained release time is calculated to be 355 h. The cumulative transdermal release of the avermectin drug is 1.15 mg, and the drug sustained release time is calculated to be 135 h.

[0197] The cumulative transdermal release of the praziquantel drug in Example 11 is 20.22 mg, and the drug sustained release time is calculated to be 161 h. The cumulative transdermal release of the pyroptidine drug is 0.80 mg, and the drug sustained release time is calculated to be 4468 h. The cumulative transdermal release of the methoprene drug is 12.33 mg, and the drug sustained release time is calculated to be 320 h. The cumulative transdermal release of the avermectin drug is 1.27 mg, and the drug sustained release time is calculated to be 122 h.

[0198] The cumulative transdermal release of the praziquantel drug in Example 12 is 21.89 mg, and the drug sustained release time is calculated to be 146 h. The cumulative transdermal release of the pyroptidine drug is 0.74 mg, and the drug sustained release time is calculated to be 4694 h. The cumulative transdermal release of the methoprene drug is 14.53 mg, and the drug sustained release time is calculated to be 267 h. The cumulative transdermal release of the avermectin drug is 1.45 mg, and the drug sustained release time is calculated to be 105 h.

[0199] The cumulative transdermal release of the praziquantel drug in Example 17 is 21.54 mg, and the drug sustained release time is calculated to be 166 h. The cumulative transdermal release of the pyroptidine drug is 1.48 mg, and the drug sustained release time is calculated to be 2628 h. The cumulative transdermal release of the methoprene drug is 23.25 mg, and the drug sustained release time is calculated to be 190 h. The cumulative transdermal release of the avermectin drug is 2.86 mg, and the drug sustained release time is calculated to be 61 h.

[0200]

[0201] a: Praziquantel b: Fipronil c: Methoxpromide d: Avermectin

[0202] Comparative analysis of the above results:

[0203] The drug permeability time of praziquantel in Example 3 is 3.37 times that of the commercially available drug, the drug permeability time of fenpropathrin is 233.21 times that of the commercially available drug, the drug permeability time of methoprene is 1.05 times that of the commercially available drug, and the drug permeability time of avermectin is 0.93 times that of the commercially available drug. It has an obvious sustained-release effect.

[0204] The drug permeability time of praziquantel in Example 8 is 142 hours less than that of Example 3 and 116 hours more than that of the commercially available drug; the drug permeability time of fenpropathrin is 1065 hours more than that of Example 3 and 6406 hours more than that of the commercially available drug; the drug permeability time of methoprene is 468 hours less than that of Example 3 and 429 hours less than that of the commercially available drug; the drug permeability time of avermectin is 155 hours less than that of Example 3 and 177 hours less than that of the commercially available drug.

[0205] The drug permeability time of praziquantel in Example 8 is 2.06 times that of the commercially available drug, the drug permeability time of fenpropathrin is 279.52 times that of the commercially available drug, the drug permeability time of methoprene is 0.45 times that of the commercially available drug, and the drug permeability time of avermectin is 0.43 times that of the commercially available drug.

[0206] The drug permeability time of praziquantel in Example 11 is 206 hours less than that of Example 3 and 52 hours more than that of the commercially available drug; the drug permeability time of fenpropathrin is 896 hours less than that of Example 3 and 4445 hours more than that of the commercially available drug; the drug permeability time of methoprene is 503 hours less than that of Example 3 and 464 hours less than that of the commercially available drug; the drug permeability time of avermectin is 168 hours less than that of Example 3 and 190 hours less than that of the commercially available drug.

[0207] The drug permeability time of praziquantel in Example 11 is 1.48 times that of the commercially available drug, the drug permeability time of fenpropathrin is 194.26 times that of the commercially available drug, the drug permeability time of methoprene is 0.41 times that of the commercially available drug, and the drug permeability time of avermectin is 0.39 times that of the commercially available drug.

[0208] The drug permeability time of praziquantel in Example 12 is 221 hours less than that of Example 3, and 37 hours more than that of the commercially available drug; the drug permeability time of fenpropathrin is 670 hours less than that of Example 3, and 4671 hours more than that of the commercially available drug; the drug permeability time of methoprene is 556 hours less than that of Example 3, and 517 hours less than that of the commercially available drug; the drug permeability time of avermectin is 185 hours less than that of Example 3, and 207 hours less than that of the commercially available drug.

[0209] The drug permeability time of praziquantel in Example 12 is 1.34 times that of the commercially available drug, the drug permeability time of fenpropathrin is 204.09 times that of the commercially available drug, the drug permeability time of methoprene is 0.34 times that of the commercially available drug, and the drug permeability time of avermectin is 0.34 times that of the commercially available drug.

[0210] The drug permeability duration of praziquantel in Example 17 is 5 hours longer than that in Example 11 and 57 hours longer than that of the commercially available drug; the drug permeability duration of fenpropathrin is 1840 hours shorter than that in Example 11 and 2605 hours longer than that of the commercially available drug; the drug permeability duration of methoprene is 130 hours shorter than that in Example 11 and 594 hours shorter than that of the commercially available drug; the drug permeability duration of avermectin is 61 hours shorter than that in Example 11 and 251 hours shorter than that of the commercially available drug.

[0211] The drug permeability time of praziquantel in Example 17 is 1.52 times that of the commercially available drug, the drug permeability time of fenpropathrin is 114.26 times that of the commercially available drug, the drug permeability time of methoprene is 0.24 times that of the commercially available drug, and the drug permeability time of avermectin is 0.20 times that of the commercially available drug.

[0212] The experimental results show that the cumulative permeation of the four drugs of Examples 3, 8, 11, 12 and 17 of the present invention is equivalent to or higher than the permeation of the corresponding drugs in the commercial products. And the penetration time of the main drug ingredients praziquantel and fepronil is higher than the commercial products, with obvious sustained release effect. Further, on the basis that the cumulative permeation of the four drugs in Example 3 is equivalent to the cumulative permeation of the commercial products, the penetration time of praziquantel, fepronil and methoprene is longer than the commercial products, and the penetration time of acetaminophen is equivalent to the commercial products. It has a good sustained release effect.

[0213] After adding HPMC, the cumulative drug permeation amount of 11 and 12 increased, which was about twice that of the commercially available products, but the drug permeation time was reduced. The permeation of different drugs in Example 17, in which crotamiton was added on the basis of Example 11, was improved to varying degrees, but the drug permeation time was significantly shortened.

[0214] In summary, Example 3 not only has good penetration, but also has obvious sustained release. The specification of the prepared compound non-pronil praziquantel gel of Example 3 is 1.2 ml / tube. It is a milky white liquid at room temperature (not higher than 25°C). It gels quickly after being dropped on the skin for 50 seconds. It has the characteristics of low oiliness and high efficiency permeability. Its sustained release effect is good, and its sustained release characteristics are better than those of commercially available preparations. The commercially available "Bolaien" drops are administered once a month, and the preparation of the present invention is expected to be administered at least once every 2-3 months, which is more convenient for medication.

[0215] The effective ingredients of the compound fepronil praziquantel gel are fepronil, methoprene, avermectin and praziquantel. Fepronil acts on ticks and flea adults as an adulticide, methoprene acts on flea eggs and larvae as an insect growth regulator, avermectin has a good expelling and killing effect on nematodes and arthropods in the body, and praziquantel has a broad-spectrum anti-schistosomiasis and anti-tapeworm effect. Therefore, the present invention can effectively prevent and treat animal fleas, tick infections, treat gastrointestinal nematode, tapeworm, hookworm and roundworm infections, and can be used as an auxiliary treatment for allergic dermatitis caused by fleas, has a stable insecticidal effect, and has a high application value.

Claims

1. A compound nonpronil praziquantel gel, characterized in that: The compound fepronil-praziquantel gel comprises fepronil, praziquantel, avermectin, methoprene, poloxamer 407, a penetration enhancer, hydroxypropyl methylcellulose, a preservative, anhydrous ethanol and water; the mass volume percentage of each component in the gel is: 6.225% of fepronil, 6.225% of praziquantel, 0.3% of avermectin, 7.5% of methoprene, 10%-13% of poloxamer 407, 6%-15% of the penetration enhancer, 0.05-0.1% of potassium sorbate, 0-1.67% of hydroxypropyl methylcellulose HPMC, 28-30% of anhydrous ethanol and the balance of water.

2. The compound nonpronil praziquantel gel according to claim 1, characterized in that: The penetration enhancer is a combination of 1,2-propylene glycol and laurocapram.

3. The compound nonpronil praziquantel gel according to claim 1 or 2, characterized in that: The mass volume percentage of each component in the compound fepronil praziquantel gel is: fepronil 6.225%, praziquantel 6.225%, avermectin 0.3%, methoprene 7.5%, poloxamer 407 10%-13%, 1,2-propylene glycol 5%-10%, laurocapram 1%-5%, potassium sorbate 0.05-0.1%, anhydrous ethanol 28-30%, and water as the balance.

4. The compound nonpronil praziquantel gel according to claim 1 or 2, characterized in that: The weight volume percentage of each component in the gel is: 6.225% of fepronil, 6.225% of praziquantel, 0.3% of acetaminophen, 7.5% of methoprene, 11%-12% of poloxamer 407, 6%-8% of 1,2-propylene glycol, 3%-5% of laurocapram, 0.05-0.1% of potassium sorbate, 28-30% of anhydrous ethanol, and the balance of water.

5. The compound non-prednisolone praziquantel gel according to any one of claims 1 to 4, characterized in that: The mass ratio of Poloxamer 407 to the total mass of 1,2-propylene glycol and laurocapram is 0.8-1.2:1, preferably 0.9-1.1:

1.

6. The compound non-prednisolone praziquantel gel according to any one of claims 1 to 5, characterized in that: The mass ratio of laurocapram to 1,2-propylene glycol is 1:1.8-1:2.2, preferably 1:2.0-2.

1.

7. The compound nonpronil praziquantel gel according to claim 1, characterized in that: The HPMC is HPMC E5 or E50, and the mass volume percentage of the HPMC is less than 0.42%.

8. The compound nonpronil praziquantel gel according to claim 7, characterized in that: When the HPMC is HPMC E50, its mass volume percentage is preferably: 0.25%-0.42%, preferably 0.25%; when the HPMC is HPMC E5, its mass volume percentage is preferably: 0.33%-0.42%, preferably 0.33%.

9. The method for preparing the compound nonpronil praziquantel gel according to claim 1, characterized in that: (1) Accurately weigh poloxamer 407, add poloxamer 407 to distilled water, seal and place in a 4°C refrigerator for 24 hours to obtain an aqueous solution of poloxamer 407; (2) adding the preservative potassium sorbate, hydroxypropyl methylcellulose and a penetration enhancer to the aqueous solution of poloxamer 407, stirring evenly, sealing and standing to obtain a matrix solution; (3) adding fenprodil, praziquantel, avermectin, and methoprene to anhydrous ethanol, dissolving them by ultrasonication, and adding the drug solution to the above-mentioned matrix solution. After stirring evenly, ultrasonication is performed in a low-temperature ultrasonic instrument until the drugs are completely dissolved, and the solution is sealed and allowed to stand; (4) Add distilled water to a certain volume and stir evenly to obtain a gel.

10. Use of the compound fepronil praziquantel gel according to any one of claims 1 to 8 in the preparation of insecticides.

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