CN115998814A - Compound tilmicosin injection and preparation method thereof - Google Patents

Abstract

The invention discloses a compound tilmicosin injection and a preparation method thereof; comprises the following components in parts by weight: 25-36 parts of tilmicosin, 1-5 parts of propranolol, 13-20 parts of coptis chinensis, 12-17 parts of radix sophorae flavescentis, 10-15 parts of mistletoe, 7-12 parts of evergreen, 7-12 parts of fomesas, 5-10 parts of liquorice, 3.5-5 parts of phosphoric acid, 18-25 parts of 1, 2-propylene glycol, 0.005-0.012 part of ethylenediamine and disodium tetraacetate; the preparation method comprises the following steps: s1, respectively crushing and mixing coptis chinensis, radix sophorae flavescentis, mistletoe, evergreen, chinese cineraria and liquorice, and extracting to obtain an extract; s2, uniformly stirring the 1, 2-propylene glycol and water for injection, adding tilmicosin, propranolol, phosphoric acid, disodium ethylenediamine tetraacetate and the extract, fully stirring, filtering, adding water for injection to fix the volume, filtering again, and sterilizing to obtain the injection; the tilmicosin can effectively relieve myocardial toxicity of tilmicosin to pigs.

Description

Compound tilmicosin injection and preparation method thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a compound tilmicosin injection and a preparation method thereof.

Background

Tilmicosin is mainly used for treating infections of animals such as actinobacillus pleuropneumoniae, pasteurella, mycoplasma, and the like, such as infectious pleuropneumonia, hemorrhagic septicemia, mycoplasma pneumonia, mammitis, swine enzootic pneumonia, infectious pleuropneumonia, and the like. The special semisynthetic macrolide antibacterial agent for tilmicosin animals has antibacterial effect similar to tylosin, has stronger activity on actinobacillus pleuropneumoniae, pasteurella and mycoplasma of livestock and poultry than tylosin, and is sensitive to 95% of haemolytic Pasteurella strains.

Tilmicosin is absorbed quickly after subcutaneous injection and is characterized by strong tissue penetration and large distribution volume (> 2L/kg). The concentration in the lung and the milk is high, the half-life period can reach 1-2 days, and the effective blood concentration is long; the cattle are subcutaneously injected according to 10mg of the weight of each 1kg, the peak concentration of blood medicine appears within 1 hour, the concentration in milk (more than 0.8 mug/ml) can be maintained for 8 to 9 days, and after one injection, lung tissue can still be maintained to be higher than the MIC of the haemolytic Pasteurella within 3 days ₉₅ (3.12 μg/ml) at a concentration of about 60:1 tilmicosin concentration in lung/serum at day 3 post injection; the concentration in milk is nearly 50 times higher than the blood concentration in 0.5 hour after subcutaneous injection, and the half life is as long as 33.8 hours.

Clinical verification shows that the pig body weight has a lethal risk by injecting 10mg/kg of tilmicosin per kilogram, and the 8mg/kg dose is a safe dose, but has larger side effects; when the common tilmicosin injection (10:3 g) is used for treatment, the safety window is narrow, so if the tilmicosin injection is directly applied to pigs, the risk of myocardial toxicity death is easy to cause, and the tilmicosin injection is mainly expressed in a cardiovascular system to cause tachycardia and weakening of contractile force. Therefore, it is important to develop a drug which can effectively treat the diseases of pigs and simultaneously can effectively relieve myocardial toxicity to pigs.

Disclosure of Invention

The application aims to overcome the defects in the prior art and provides a compound tilmicosin injection and a preparation method thereof; by adding the traditional Chinese medicine components and propranolol into the existing tilmicosin injection, the heart rate of pigs can be kept stable, and further the toxicity of tilmicosin on cardiovascular and heart is relieved; after scientific compatibility, the product can be directly used for treating respiratory diseases of pigs caused by infection of actinobacillus pleuropneumoniae, pasteurella, mycoplasma and the like, and can effectively relieve toxicity of the pigs during treatment of diseases.

The technical aim of the invention is realized by the following technical scheme:

a compound tilmicosin injection comprises the following components in parts by weight:

25-36 parts of tilmicosin, 1-5 parts of propranolol, 13-20 parts of coptis chinensis, 12-17 parts of radix sophorae flavescentis, 10-15 parts of mistletoe, 7-12 parts of evergreen, 7-12 parts of fomesas, 5-10 parts of liquorice, 3.5-5 parts of phosphoric acid, 18-25 parts of 1, 2-propylene glycol and 0.005-0.012 part of disodium ethylenediamine tetraacetate.

In the scheme, the effects of the Chinese herbal medicine and propranolol in the invention are as follows:

coptis root: contains berberine, and has antiarrhythmic, peripheral resistance reducing, heart rate slowing, and blood pressure lowering effects; the Berberine (BER) and its derivatives contained in Coptidis rhizoma can increase superoxide dismutase (SOD) activity in brain of cerebral ischemia animal, reduce Malondialdehyde (MDA) content, inhibit leakage of Lactic Dehydrogenase (LDH) and MDA content of cultured nerve cells, promote reduction of Glutathione (GSH), increase insulin bioactivity, increase liver SOD activity, reduce liver lipid peroxidation level, and improve organism antioxidant capacity; rhizoma Coptidis also has immunity regulating, antiinflammatory, antiviral, and antitumor effects;

radix Sophorae Flavescentis: has obvious heart inhibiting effect, and can effectively slow down heart rate, weaken myocardial contractility, reduce cardiac output, and also has antiarrhythmic, antibacterial, antiinflammatory, antitumor, leukocyte increasing, asthma relieving, phlegm eliminating, tranquilization, antiallergic, immunosuppression, etc.;

loranthus mulberry mistletoe: has diuretic, antihypertensive, antiviral, antiarrhythmic, acute myocardial infarction preventing and treating, platelet aggregation inhibiting, and isolated cardiac coronary flow increasing effects; the activity of superoxide dismutase and glutathione peroxidase can be obviously improved;

rohdea japonica: can increase myocardial contractility, dilate coronary artery to slow heart rate, and improve microcirculation and coronary circulation;

herb of Chinese Thunberg shou: has effects in tonifying heart, promoting urination, tranquilizing mind, and slowing down heart rate, and can be used for reducing nervous system excitability and spinal cord reflex hyperfunction, and for treating acute and chronic cardiac insufficiency;

licorice root: has effects in stabilizing heart rate and relieving drug properties;

propranolol: can be used for treating arrhythmia caused by various reasons, such as atrial and ventricular premature beat (with good effect), sinus and supraventricular tachycardia, and atrial fibrillation.

The synergy between the above drugs is as follows:

when the international tilmicosin injection is used for pigs, the activities of superoxide dismutase and glutathione peroxidase in myocardial tissues can be reduced, so that myocardial cells are apoptotic, and tachycardia and heart expansion can be caused, so that myocardial toxic death is caused, and the international tilmicosin injection is unfavorable for clinical use; on the basis, propranolol is added, so that the effects of slowing down heart rate and reducing myocardial oxygen consumption can be rapidly exerted, and the effect is rapid; however, if propranolol is compounded with tilmicosin only, the compounded drug has at least four defects:

firstly, the propranolol has a high metabolism speed and a half-life period of 2-3 hours, so that the drug effect can be maintained by taking the propranolol for many times, and excessive tilmicosin is taken by pigs by taking the propranolol for many times, so that toxicity is increased;

secondly, the propranolol has an excessively fast drug effect, and has a good effect of reducing heart rate, so that the drug effect is often excessive, and further symptoms such as hypotension, bradycardia and the like are caused;

thirdly, the compound medicine can not improve the activity of superoxide dismutase and glutathione peroxidase, so that myocardial cells still can continuously die, and the toxic and side effects are not reduced;

fourth, it is: the long-term use of the compound medicine can lead to low immunity of pigs and cause various allergic reactions;

based on the above, the compound medicine is difficult to be directly applied to pig therapy, on the basis, coptis chinensis, mistletoe, evergreen, radix sophorae flavescentis and foggy mane are added into the compound medicine, and by adding coptis chinensis and mistletoe, the activity of superoxide dismutase and glutathione peroxidase can be improved, the toxicity of tilmicosin to myocardial cells is reduced, and simultaneously the coptis chinensis and the mistletoe also have the effect of reducing heart rate; the toxicity of the drug compounded by tilmicosin and propranolol can reduce the blood flow of heart coronary arteries, and the addition of Rohdea japonica can increase the blood flow of coronary arteries, induce urination and prevent myocardial infarction, so that the survival rate of pigs is improved; by adding the kuh-seng, the effects of soothing the nerves and resisting allergy can be achieved; by adding the foggy mane, the central nerve can be protected, excitability of a nervous system and spinal cord hyperreflexia can be reduced, and adverse effects of toxic and side effects of medicines on the central nerve can be further reduced, and meanwhile, the foggy mane also has the effects of promoting urination and strengthening heart; through the combined action of the evergreen and the fomes officinalis, the diuretic effect of the medicine can be improved, the metabolism of the medicine is accelerated, and the residue of the medicine in the body is reduced; furthermore, the coptis chinensis, the mistletoe, the evergreen, the kuh-seng and the foggy manyflower all have the effect of reducing the heart rate, so that the device can play a role of reducing the heart rate together with the propranolol, and the problem that the propranolol is excessive in efficacy, low in blood pressure or slow in heart rate and dead is caused can be effectively relieved. However, in actual use, the compatibility of the medicines is not added, and the medicines after compounding can slow down the problem of excessive medicine effect, but the lowered heart rate is difficult to continuously stabilize, and the problem of bradycardia still occurs. By adding the 6 Chinese herbal medicines on the basis of tilmicosin and propranolol, the effect timeliness of the compound injection can be greatly prolonged.

Through the interaction among the medicines, the compound tilmicosin injection can be directly used for treating respiratory diseases of pigs caused by infection of actinobacillus pleuropneumoniae, pasteurella, mycoplasma and the like, and can effectively relieve toxicity of the compound tilmicosin injection to the heart and the cardiovascular diseases of the pigs during the treatment of the diseases; specifically, the injection can improve the activities of superoxide dismutase and glutathione peroxidase, and reduce the toxicity of tilmicosin to myocardial cells; the heart rate can be effectively reduced, and the reduced heart rate can be ensured to be continuously and stably maintained; can also effectively prolong the drug effect of the injection, and has the functions of enhancing the immunity of pigs, reducing anaphylactic reaction and the like.

Preferably, the injection comprises the following components in parts by weight: 28-35 parts of tilmicosin, 2-4 parts of propranolol, 15-18 parts of coptis chinensis, 13-15 parts of radix sophorae flavescentis, 12-14 parts of mistletoe, 8-11 parts of evergreen, 8-10 parts of fomesas, 6-9 parts of liquorice, 4-4.5 parts of phosphoric acid, 20-23 parts of 1, 2-propylene glycol and 0.007-0.01 part of disodium ethylenediamine tetraacetate. In the scheme, the toxic and side effects of the injection on pigs can be further reduced by further limiting the dosage of each component in the injection.

Preferably, the injection comprises the following components in parts by weight: 33 parts of tilmicosin, 3 parts of propranolol, 16 parts of coptis chinensis, 14 parts of radix sophorae flavescentis, 13.5 parts of mistletoe, 10 parts of evergreen, 8.5 parts of fomesas, 7 parts of liquorice, 4.3 parts of phosphoric acid, 21 parts of 1, 2-propylene glycol and 0.009 part of disodium ethylene diamine tetraacetate. In the scheme, the toxic and side effects of the injection on pigs can be further reduced by further limiting the dosage of each component in the injection.

In addition, in order to achieve the aim, the invention also provides a preparation method of the compound tilmicosin injection, which comprises the following steps:

s1, respectively crushing and mixing coptis chinensis, radix sophorae flavescentis, mistletoe, evergreen, chinese cineraria and liquorice, and extracting to obtain an extract;

s2, uniformly stirring the 1, 2-propylene glycol and water for injection, then adding tilmicosin, propranolol, phosphoric acid, disodium ethylenediamine tetraacetate and the extract, fully stirring, then filtering, adding water for injection, fixing the volume, and filtering again. Sterilizing to obtain the injection.

In the scheme, the extraction efficiency of the coptis chinensis, the radix sophorae flavescentis, the mistletoe, the evergreen, the Chinese cineraria and the liquorice are improved by crushing; in the extraction process, the active ingredients of the Chinese herbal medicines can be fully dissolved out, so that the drug effect of the injection is improved; and S2, filtering is carried out twice successively, so that impurities in the injection can be effectively reduced, the transparency and clarity of the injection are improved, and further the injection is ensured to exert the drug effect smoothly.

Preferably, the step S1 includes the following steps:

s11, respectively crushing and mixing the coptis chinensis, the kuh-seng, the mistletoe, the evergreen, the foggy mane and the liquorice, then adding water with the mass being 7-8 times that of the mixture, soaking for 20-40min, and then heating to slight boiling and keeping for 80-100min; filtering to obtain a first filtrate and filter residues;

s12, adding water with the mass which is 5-7 times that of the filter residue, heating to slight boiling, maintaining for 50-70min, and filtering to obtain a second filtrate;

and S13, combining and concentrating the first filtrate and the second filtrate, adding an ethanol solution after cooling, fully stirring, standing, filtering, and concentrating, standing and re-concentrating the filtrate to obtain the extract.

In the above scheme, water extraction and alcohol precipitation can be carried out to further remove water-soluble impurities such as starch, gum, pectin, mucilaginous substance, protein, ravage, pigment, inorganic salt, etc., and purify the medicinal liquid.

Preferably, the crushed Chinese herbal medicine in the step S11 is mixed after being sieved by a 50-70 mesh sieve.

Preferably, the filtrate after meal rest in step S13 needs to be adjusted to pH 7.8-8.2 before concentrating the extract.

Preferably, during the re-filtration in step S2, the filtration is performed through 0.45 μm and then through 0.22 μm filter pores.

In the scheme, impurities in the injection can be further removed by filtering the filtrate twice, so that the transparency and clarity of the injection are further enhanced, and the injection is ensured to be qualified.

Compared with the prior art, the invention has the beneficial effects that:

through the interaction among the medicines, the compound tilmicosin injection can be directly used for treating respiratory diseases of pigs caused by infection of actinobacillus pleuropneumoniae, pasteurella, mycoplasma and the like, and can effectively relieve toxicity of the compound tilmicosin injection to the heart and the cardiovascular diseases of the pigs during the treatment of the diseases; specifically, the injection can improve the activities of superoxide dismutase and glutathione peroxidase, and reduce the toxicity of tilmicosin to myocardial cells; the heart rate can be effectively reduced, and the reduced heart rate can be ensured to be continuously and stably maintained; can also prolong the action time of the injection, strengthen the immunity of pigs, reduce allergic reaction and the like.

Detailed Description

The following will clearly and fully describe the technical solutions in the embodiments of the present application, and it is obvious that the described embodiments are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art based on the embodiments herein without making any inventive effort, are intended to be within the scope of the present application.

Example 1

S11, respectively screening impurities which cannot be used as medicines, such as cores, stems, bones, shells and the like, or deteriorated and invalid parts, such as worm-eaten and mildewed parts, in coptis chinensis, radix sophorae flavescentis, herba taxilli 10g, herba orostachyos 12g, herba orostachyos 11g and liquorice 9g, respectively crushing the selected coptis chinensis 18g, radix sophorae flavescentis 15g, herba orostachyos 10g, herba orostachyos 12g and liquorice 9g, sieving the crushed materials with a 60-mesh sieve, fully mixing the crushed materials to prepare a mixture, and pouring the mixture into an extraction tank; then adding water which is 7 times of the total mass of the mixture, soaking for 25min, heating to micro-boiling and keeping the micro-boiling state for 90min; then the decoction is filtered by a 80-mesh filter screen to obtain a first filtrate and filter residues;

s12, adding 6 times of water into the filter residue, heating to slight boiling, keeping the slight boiling state for 70min, and then filtering the decoction through a 80-mesh filter screen to obtain a second filtrate;

s13, adding the first filtrate and the second filtrate into a liquid storage tank for combination, standing for 2 hours, inputting the supernatant into a double-effect concentration tank, starting a steam valve for reduced pressure concentration (the temperature is 70-80 ℃, the vacuum degree is 0.02-0.04 Mpa), and concentrating the filtrate to 97.5-100g; transferring the concentrated thick paste into an alcohol precipitation tank, cooling to below 60 ℃, slowly adding 95% ethanol to ensure that the ethanol content is more than 60%, fully stirring, standing and refrigerating for 12 hours, filtering, concentrating the filtrate under reduced pressure to 45-52g (the vapor pressure is not more than 0.09Mpa and the vacuum degree is minus 0.06 Mpa), standing for 12 hours in a cold meal, filtering, regulating the pH value of the filtrate to 8 by using an ammonia test solution, stirring for 7 minutes, standing for 6 hours, and filtering; then ethanol distillation recovery is carried out on the filtrate,

concentrating the filtrate until no ethanol smell exists, to obtain extract;

s2, stirring 22g of 1, 2-propylene glycol and a small amount of water for injection for 10min, adding 34g of tilmicosin, 3g of propranolol, 3.5g of phosphoric acid, 0.012g of disodium ethylenediamine tetraacetate and the extract, stirring for 15min, filtering, adding water for injection to a volume of 100ml, filtering by a 0.45 mu m and 0.22 mu m microporous cylinder type filter membrane filter, and sterilizing to obtain the injection.

Example 2

Compared with the embodiment 1, the injection has the following components:

30g of tilmicosin, 3.5g of propranolol, 15g of coptis chinensis, 15g of radix sophorae flavescentis, 12.5g of mistletoe, 10g of Rohdea japonica, 9g of fomesangial, 7g of liquorice, 4.5g of phosphoric acid, 20g of 1, 2-propanediol and 0.008g of disodium ethylene diamine tetraacetate.

The remaining steps and parameters were the same as in example 1.

Example 3

Compared with the embodiment 1, the injection has the following components:

33g of tilmicosin, 3g of propranolol, 16g of coptis chinensis, 14g of radix sophorae flavescentis, 13.5g of mistletoe, 10g of evergreen, 8.5g of fossa shou, 7g of liquorice, 4.3g of phosphoric acid, 21g of 1, 2-propanediol and 0.009g of disodium ethylene diamine tetraacetate.

The remaining steps and parameters were the same as in example 1.

Example 4

S11, respectively screening impurities which cannot be used as medicines, such as cores, stems, bones, shells and the like, or deteriorated and invalid parts, such as worm-eaten and mildewed parts, in coptis chinensis, radix sophorae flavescentis, herba taxilli 10g, herba orostachyos 12g, herba orostachyos 11g and liquorice 9g, respectively crushing the selected coptis chinensis 18g, radix sophorae flavescentis 15g, herba orostachyos 10g, herba orostachyos 12g and liquorice 9g, sieving the crushed materials with a 60-mesh sieve, fully mixing the crushed materials to prepare a mixture, and pouring the mixture into an extraction tank; then adding water which is 8 times of the total mass of the mixture, soaking for 20-40min, heating to micro-boiling and keeping the micro-boiling state for 100min; then the decoction is filtered by a 80-mesh filter screen to obtain a first filtrate and filter residues;

s12, adding water with the mass which is 5 times that of the filter residue, heating to slight boiling, keeping the slight boiling state for 70min, and then filtering the decoction through a 80-mesh filter screen to obtain second filtrate;

s13, adding the first filtrate and the second filtrate into a liquid storage tank for combination, standing for 2 hours, inputting the supernatant into a double-effect concentration tank, starting a steam valve for reduced pressure concentration (the temperature is 70-80 ℃, the vacuum degree is 0.02-0.04 Mpa), and concentrating the filtrate to 97.5-100g; transferring the concentrated thick paste into an alcohol precipitation tank, cooling to below 60 ℃, slowly adding 95% ethanol to ensure that the ethanol content is more than 60%, fully stirring, standing and refrigerating for 12 hours, filtering, concentrating the filtrate under reduced pressure to 45-52g (the vapor pressure is not more than 0.09Mpa and the vacuum degree is minus 0.06 Mpa), standing for 12 hours in a cold meal, filtering, regulating the PH value of the filtrate to 8.2 by using an ammonia test solution, stirring for 10 minutes, standing for 6 hours, and filtering; then, ethanol distillation recovery is carried out on the filtrate, and sterilization is carried out after the filtrate is concentrated until no ethanol smell exists, so as to obtain extractum;

s2, stirring 22kg of 1, 2-propylene glycol and a small amount of water for injection for 15min, adding 34g of tilmicosin, 3g of propranolol, 3.5g of phosphoric acid, 0.012g of disodium ethylenediamine tetraacetate and the extract, stirring for 10min, filtering, adding water for injection to constant volume of 100ml, filtering by a 0.45 mu m and 0.22 mu m microporous cylinder type filter membrane filter, and sterilizing to obtain injection.

Comparative example 1

10% tilmicosin injection (this is prior art).

Comparative example 2

22g of 1, 2-propanediol, 34g of tilmicosin, 3g of propranolol, 0.012g of disodium ethylenediamine tetraacetate, 3.5g of phosphoric acid and a small amount of water for injection are uniformly stirred for 10min, then the water for injection is added to a volume of 100ml, and then the solution is filtered by a microporous cylinder type filter membrane filter with 0.45 mu m and 0.22 mu m, and the injection is obtained after sterilization.

Comparative example 3

S11, respectively screening impurities which cannot be used as medicines, such as cores, stems, peduncles, bones, shells and the like, or deteriorated and invalid parts, such as worm-eaten and mildewed parts, in the kuh-seng, the evergreen 12g, the evergreen 11g and the liquorice 9g, respectively crushing the selected kuh-seng, the evergreen 12g, the evergreen 11g and the liquorice 9g, sieving the crushed materials with a 60-mesh sieve, fully mixing the crushed materials to prepare a mixture, and pouring the mixture into an extraction tank; then adding water which is 7 times of the total mass of the mixture, soaking for 25min, heating to micro-boiling and keeping the micro-boiling state for 90min; then the decoction is filtered by a 80-mesh filter screen to obtain a first filtrate and filter residues;

s12, adding 6 times of water into the filter residue, heating to slight boiling, keeping the slight boiling state for 70min, and then filtering the decoction through a 80-mesh filter screen to obtain a second filtrate;

s13, adding the first filtrate and the second filtrate into a liquid storage tank for combination, standing for 2 hours, inputting the supernatant into a double-effect concentration tank, starting a steam valve for reduced pressure concentration (the temperature is 70-80 ℃, the vacuum degree is 0.02-0.04 Mpa), and concentrating the filtrate to 70g; transferring the concentrated thick paste into an alcohol precipitation tank, cooling to below 60 ℃, slowly adding 95% ethanol to enable the alcohol content to be more than 60%, fully stirring, standing and refrigerating for 12 hours, filtering, concentrating the filtrate under reduced pressure to 30kg (steam pressure: not more than 0.09Mpa and vacuum degree: -0.06 Mpa), standing for 12 hours in a cold meal, filtering, regulating the PH value of the filtrate to 8 by using ammonia test solution, stirring for 7 minutes, standing for 6 hours, and filtering; then, ethanol distillation recovery is carried out on the filtrate, and sterilization is carried out after the filtrate is concentrated until no ethanol smell exists, so as to obtain extractum;

s2, uniformly stirring a small amount of water for injection of 22g of 1, 2-propylene glycol for 10min, adding 34g of tilmicosin, 3g of propranolol, 3.5g of phosphoric acid, 0.012g of disodium ethylenediamine tetraacetate and the extract, stirring for 15min, filtering, adding water for injection to 100ml, filtering by a 0.45 mu m and 0.22 mu m microporous cylinder type filter membrane filter, and sterilizing to obtain injection.

Experimental example 1 stability test

(1) High temperature test

Taking the injection solutions in examples 1-4 and comparative examples 1-3, respectively placing at 60 ℃ for 10 days, and sampling and detecting related indexes on the 5 th day and the 10 th day; the results show that all the injections do not change significantly and meet the injection standard.

(2) Strong light irradiation test

Taking the injection solutions in examples 1-4 and comparative examples 1-3, placing the injection solutions in an illumination box with a fluorescent lamp and under the condition of 4500+/-500 lx illuminance for 10 days, sampling and detecting on the 5 th day and the 10 th day, wherein the results show that the content and the properties have no obvious difference, and the injection solutions meet the injection standard.

(3) Acceleration test

Taking the injection solutions in examples 1-4 and comparative examples 1-3, and placing the injection solutions under the conditions of (40+/-2) DEG C and RH (75+/-5)%, and performing a 6-month test; sampling and detecting at the end of the 0 th, 1 st, 2 nd, 3 rd and 6 th months in the test period, and the results show that the content and the properties have no obvious difference and accord with the injection standard.

Experimental example 2 toxic side Effect test

Three-day pig farm piglets were taken and randomly divided into 5 groups (designated as groups 1-5), 10 heads per group, and the injections in examples 1-4 and no drug (blank group) were injected, respectively. Groups 1-4 were all injected with one needle on days 3, 7 and 21, and the total condition of the piglets was observed after 50 days, and the incidence of respiratory diseases was counted, based on tilmicosin, and the results are shown in table 1.

TABLE 1

	Incidence (%)
		Group 1	0％
Group 2	0％
		Group 3	0％
Group 4	0％
		Group 5	30％

As can be seen from table 1: after the medicine (group 1-4) is injected, pigs have no adverse reaction, namely the injection has almost no toxic or side effect on pigs, and can well prevent respiratory diseases, in addition, the medicine is injected into the muscle according to 12mg/kg of tilmicosin, and has no toxic or side effect, compared with tilmicosin solution (namely comparative example 1) in the prior art, the injection has only 10mg/kg of toxic or side effect on pigs, and has great progress. In addition, during 60 days, only 20% of the 40 pigs in the 1-4 groups have cold, and the 5 pigs have low resistance due to no drug injection, and more than 40% of the pigs have cold symptoms, so that the injection can effectively improve the immunity of the pigs and improve the capability of resisting cold viruses.

Experimental example 3 clinical test of therapeutic Effect on respiratory diseases

160 pigs in a pig farm with an outbreak of infectious pleuropneumonia were taken, the time for each pig to develop symptoms was less than 36 hours, after which the pigs were equally divided into 8 groups, and the injections of examples 1-4, comparative example 1 (water for injection added to the total), comparative example 2 and comparative example 3 were respectively injected, wherein the injections of examples 1-4 and comparative examples 2-3 were all intramuscular injections of tilmicosin at 12mg/kg body weight, the injection of comparative example 1 was intramuscular injections of tilmicosin at 10mg/kg body weight, the injections were continued once a day, and after 5 days the death of the pigs was observed, and the results are shown in table 2.

TABLE 2

As can be seen from Table 2, the injection in examples 1-4 has a good therapeutic effect on diseased pigs, the mortality rate is less than or equal to 10%, and the rest of non-dead pigs are recovered or in recovery stage; in the comparative example 1, only tilmicosin injection is adopted for treatment, and the treatment effect is effective, but the tilmicosin injection has stronger side effect, so that pigs are easy to die; the injection in comparative example 2 is not added with Chinese herbal medicine, and the injection in comparative example 3 is not strictly prepared according to the proportion and the dosage, so that the treatment effect is relatively poor, and the synergistic effect among medicines is reflected.

In addition, the elimination half-life of propranolol in the pig was examined after intramuscular injection of the pig in experimental example 3, and the examination results are shown in table 3 (wherein half-life is an average value).

TABLE 3 Table 3

	Half-life (h)
		Example 1	3.5
Example 2	3.75
		Example 3	4
Example 4	3.6
		Comparative example 2	2.8
Comparative example 3	3

As can be seen from table 3: the injection provided by the invention can obviously improve the half-life period of propranolol drug, so that the drug effect is prolonged; meanwhile, the Chinese herbal medicine also has the effect of reducing heart rate, so that the defect of reduced efficacy can be overcome by the effect of other Chinese herbal medicines even if the efficacy of propranolol is reduced, and further the heart rate can be effectively stabilized and the toxicity of tilmicosin can be reduced in the injection period of tilmicosin.

Further, in the intramuscular injection period of the pig in the experimental example 3, the heart rate and the blood pressure of the pig are continuously monitored, the specific monitoring time is 1,2, 3 and 4 … … hours after the injection, each time is monitored for 5 minutes, and the experimental result shows that the heart rate and the blood pressure of the pig injected with the injection in the examples 1-4 are kept in a normal range and the numerical value is stable; the heart rate of pigs in comparative example 1 is generally higher than the normal range; the heart rate of comparative example 2 generally fluctuates in the normal range and a range slightly lower than the normal range within the first 5 hours, whereas after 7 hours, a part of the heart rate of pigs appears to be higher than the normal range; the heart rate of comparative example 3 generally fluctuates in a normal range and a range slightly lower than the normal range, and the heart rate of part of pigs is higher than the normal range within more than 15 hours; therefore, the injection in the invention can also keep the heart rate of pigs within a normal range, thereby reducing the toxicity to cardiac muscle.

The foregoing is merely a preferred embodiment of the invention, and it is to be understood that the invention is not limited to the form disclosed herein and is not to be considered as excluding other embodiments, but is capable of numerous other combinations, modifications and environments and is capable of modifications within the scope of the inventive concept, either by the foregoing teachings or by the teaching of the relevant art. And that modifications and variations which do not depart from the spirit and scope of the invention are intended to be within the scope of the appended claims.

Claims (8)

1. The compound tilmicosin injection is characterized by comprising the following components in parts by weight:

25-36 parts of tilmicosin, 1-5 parts of propranolol, 13-20 parts of coptis chinensis, 12-17 parts of radix sophorae flavescentis, 10-15 parts of mistletoe, 7-12 parts of evergreen, 7-12 parts of fomesas, 5-10 parts of liquorice, 3.5-5 parts of phosphoric acid, 18-25 parts of 1, 2-propylene glycol and 0.005-0.012 part of disodium ethylenediamine tetraacetate.

2. The compound tilmicosin injection as claimed in claim 1, which is characterized by comprising the following components in parts by weight: 28-35 parts of tilmicosin, 2-4 parts of propranolol, 15-18 parts of coptis chinensis, 13-15 parts of radix sophorae flavescentis, 12-14 parts of mistletoe, 8-11 parts of evergreen, 8-10 parts of fomesas, 6-9 parts of liquorice, 4-4.5 parts of phosphoric acid, 20-23 parts of 1, 2-propylene glycol and 0.007-0.01 part of disodium ethylenediamine tetraacetate.

3. The compound tilmicosin injection as claimed in claim 1, which is characterized by comprising the following components in parts by weight: 33 parts of tilmicosin, 3 parts of propranolol, 16 parts of coptis chinensis, 14 parts of radix sophorae flavescentis, 13.5 parts of mistletoe, 10 parts of evergreen, 8.5 parts of fomesas, 7 parts of liquorice, 4.3 parts of phosphoric acid, 21 parts of 1, 2-propylene glycol and 0.009 part of disodium ethylene diamine tetraacetate.

4. A method for preparing a compound tilmicosin injection according to any one of claims 1-3, comprising the steps of:

s1, respectively crushing and mixing coptis chinensis, radix sophorae flavescentis, mistletoe, evergreen, chinese cineraria and liquorice, and extracting to obtain an extract;

s2, uniformly stirring the 1, 2-propylene glycol and water for injection, then adding tilmicosin, propranolol, phosphoric acid, disodium ethylenediamine tetraacetate and the extract, fully stirring, and then filtering, adding water for injection to fix the volume, filtering again and sterilizing to obtain the injection.

5. The method for preparing the compound tilmicosin injection according to claim 4, wherein the step of S1 is as follows:

s11, respectively crushing and mixing the coptis chinensis, the kuh-seng, the mistletoe, the evergreen, the foggy mane and the liquorice, then adding water with the mass being 7-8 times that of the mixture, soaking for 20-40min, and then heating to slight boiling and keeping for 80-100min; filtering to obtain a first filtrate and filter residues;

s12, adding water with the mass which is 5-7 times that of the filter residue, heating to slight boiling, maintaining for 50-70min, and filtering to obtain a second filtrate;

and S13, combining and concentrating the first filtrate and the second filtrate, adding an ethanol solution after cooling, fully stirring, standing, filtering, and concentrating, standing and re-concentrating the filtrate to obtain the extract.

6. The method for preparing a compound tilmicosin injection according to claim 5, wherein the crushed Chinese herbal medicines in the step S11 are mixed after passing through a 50-70 mesh sieve.

7. The method for preparing a compound tilmicosin injection according to claim 5, wherein the filtrate after meal rest in step S13 is adjusted to pH 7.8-8.2, and then the extract is concentrated.

8. The method for preparing a compound tilmicosin injection according to claim 4, wherein in the step S2, the compound tilmicosin injection is filtered through a 0.45 μm filter and a 0.22 μm filter successively.