CN111437249A

CN111437249A - Compound gentamicin sulfate injection and preparation method and application thereof

Info

Publication number: CN111437249A
Application number: CN202010317944.7A
Authority: CN
Inventors: 廖雪玲; 徐玉明
Original assignee: Shanghai Gongyi Pharmaceutical Co ltd
Current assignee: Shanghai Gongyi Pharmaceutical Co ltd
Priority date: 2020-04-21
Filing date: 2020-04-21
Publication date: 2020-07-24
Anticipated expiration: 2040-04-21
Also published as: CN111437249B

Abstract

The invention discloses a compound gentamicin sulfate injection which is characterized by comprising the following components in percentage by weight: gentamicin sulfate 10%; 5% of lincomycin hydrochloride; a synergist: 1% of trimethoprim; antioxidant: 0.2 percent of sodium metabisulfite; a stabilizer: 2% of sodium citrate; solvent: propylene glycol 20%, the rest is water for injection, and citric acid is adopted to adjust the pH value to 5.0-6.0. The invention also discloses a preparation method and application thereof. The compound injection has stronger killing effect on haemophilus parasuis and streptococcus suis, improves the killing effect and reduces the treatment cost by combining the existing antibiotic drugs.

Description

Compound gentamicin sulfate injection and preparation method and application thereof

Technical Field

The invention relates to the field of antibiotic preparation, and in particular relates to a compound gentamicin sulfate injection as well as a preparation method and application thereof.

Background

The porcine respiratory disease syndrome refers to a series of diseases with characteristics of slow growth speed, high feed-meat ratio, anorexia, cough, asthma, dyspnea and the like caused by combined action of pathogens of pigs in a certain stress environment.

At present, the porcine respiratory disease syndrome is mainly caused by mixed infection of primary pathogen and secondary pathogen. Wherein secondary infection caused by streptococcus suis, haemophilus parasuis and the like has the characteristics of high elimination rate and high death rate.

Investigation shows that the bacterial respiratory disease of the pig mostly occurs in winter and spring, and the bacterial respiratory disease is a common frequently-occurring disease in China at present. The disease is susceptible to the pigs in the nursing stage and the fattening stage (6-22 weeks old), and the affected pigs mainly have the symptoms of loss of appetite, heat, asthma, increase of nasal cavity and eye secretion, acceleration of respiration and the like. The incidence of bacterial respiratory disease in pigs is reported to be high, mortality is low, and mortality tends to decline with the age of the affected pigs. But the recovered pigs almost lose the production value, and the economic loss of the pig bacterial respiratory disease to the live pig breeding industry is very disastrous.

Harm of porcine bacterial respiratory disease syndrome the death of pigs suffering from bacterial disease syndrome suffers from huge economic losses for breeders. The sick pig is infectious and threatens the health of other healthy pigs. The growth of the recovered pigs is inhibited and the development is slow, the slaughtering time is prolonged, the feed-weight ratio is increased, and the comprehensive cost of cultivation is increased.

In addition, some zoonosis exists, which threatens the health and even life of human beings. Such as Streptococcus suis, although not common in humans, the condition is severe and can even be fatal. At present, 60 countries report that nearly thousands of cases infected with streptococcus suis are accumulated, and some infected cases exist in China.

Many bacteria have developed resistance to antibiotics due to abuse problems, and even multi-resistant strains have emerged.

Therefore, it is very critical how to use the existing antibacterial drugs to slow down the generation of bacterial durability while increasing the therapeutic efficiency of the disease.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention aims to provide a compound gentamicin sulfate injection.

The second purpose of the invention is to provide a preparation method of the compound gentamicin sulfate injection.

The invention also aims to provide the application of the compound gentamicin sulfate injection.

In order to realize one of the purposes of the invention, the adopted technical scheme is as follows:

a compound gentamicin sulfate injection comprises the following components in percentage by weight:

gentamicin sulfate 10%;

5% of lincomycin hydrochloride;

a synergist: 1% of trimethoprim;

antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: 20% of propylene glycol;

the balance of water for injection, and the pH is adjusted to 5.0-6.0 by citric acid.

In order to realize the second purpose of the invention, the adopted technical scheme is as follows:

a preparation method of compound gentamicin sulfate injection comprises the following steps:

adding the sodium metabisulfite and the sodium citrate into the water for injection, and stirring for dissolving;

adding the propylene glycol into the trimethoprim, heating to 50-60 ℃, and stirring for dissolving;

mixing the two solutions, sequentially adding the gentamicin sulfate and the lincomycin hydrochloride, and stirring for dissolving;

adjusting the pH to 4.5-6.5 with the citric acid;

adding water to the total amount of the preparation, filtering, filling nitrogen, filling and sealing, and sterilizing at 121 ℃ for × 15 min.

In order to realize the third purpose of the invention, the technical scheme is as follows:

the compound gentamicin sulfate injection is used for preparing a medicament for treating streptococcus suis infection.

In a preferred embodiment of the invention, said treatment of streptococcosis suis comprises treatment of infection by streptococcus suis or haemophilus parasuis.

The invention has the beneficial effects that:

the compound injection has stronger killing effect on haemophilus parasuis and streptococcus suis, improves the killing effect and reduces the treatment cost by combining the existing antibiotic drugs.

Detailed Description

The noun explains:

streptococcosis suis disease

Streptococcus suis is a zoonotic disease caused by a variety of pathogenic Streptococcus suis infections. Streptococcus suis is a common and important pathogen in pigs and also a common cause of meningitis of human and animal origin, and can cause meningitis, septicemia, endocarditis, arthritis and pneumonia, mainly manifested as fever and severe toxemia symptoms.

The secondary infection lethality caused by streptococcus suis, haemophilus parasuis and the like is high, and the harm is serious, so that the primary task of preventing and treating the porcine respiratory disease is to solve the problem of secondary infection caused by gram-positive bacteria streptococcus suis and gram-negative haemophilus parasuis. The most common pathogenic bacteria isolated from the respiratory system of pigs with clinical symptoms in the laboratory are streptococcus suis, haemophilus parasuis, actinobacillus pleuropneumoniae and the like. The detection rate of Mycoplasma hyopneumoniae is also very high.

The respiratory system of the pig has three lines of defense to exogenous foreign matters, and the three lines of defense are mainly used for timely removing pathogens, generating immune response and recovering the health of the pig. Mucociliary and special turbinate structure inside the pig nose can moisten and filter air entering the nose, which is a first line of defense; adhesion of bronchial cilia to pathogens, which is the second line of defense; the third line of defense is to carry out the elimination of pathogenic bacteria and immune response. The three defense lines can clean and filter exogenous impurities, viruses and bacteria together, so that the health of the pigs is ensured. When the pig body suffers from pathogenic factors such as pathogenic bacteria invasion or body resistance reduction, the respiratory system of the susceptible pig group is easily damaged by other pathogenic bacteria invasion, so that serious respiratory system diseases occur.

Gentamicin sulfate injection

Gentamicin sulfate is aminoglycoside antibiotic and has excellent antibacterial effect on gram-positive and gram-negative bacteria. The action mechanism of gentamicin sulfate is combined with a bacterial ribosome 30S subunit, the synthesis of bacterial protein is inhibited, the defect of bacterial cell membranes and cell walls is indirectly caused, the action of the gentamicin sulfate on killing bacteria in the stationary phase is strong, the gentamicin sulfate has an antibiotic after effect on a plurality of pathogenic bacteria, and the gentamicin sulfate belongs to a class 2 stationary phase bactericide in the classification of antibacterial drugs.

The gentamicin sulfate has wide antibacterial spectrum, low price, mature process and stable property, so the gentamicin sulfate is widely used in veterinary clinic. The gentamicin sulfate injection is mainly injected intramuscularly for 0.5h

The peak concentration of the blood medicine can be reached in a short time, and the absorption is rapid and complete. The peak concentration of the blood drug was slightly delayed upon subcutaneous injection. After absorption, it is mainly distributed in extracellular fluid, and the drugs can appear in ascites, pleural fluid, pus and synovial fluid. Can permeate placenta tissue, and has concentration of 15-50% of that in mother body.

The metabolism of gentamicin sulfate is mainly excreted through glomeruli by urine, and accounts for about 40-80% of the dosage. Is mainly used for infection caused by sensitive bacteria such as streptococcus, escherichia coli, proteus and the like. Such as infections of the intestinal tract, respiratory tract, urinary tract, soft tissues, etc. Tests show that the curative effect of treating pullorum disease and chicken pasteurellosis by using gentamicin sulfate is obvious. The effect of Jipei Hua, etc. on treating bacillary dysentery is compared with that of norfloxacin and gentamicin sulfate, and the comparison between norfloxacin and gentamicin sulfate (p <0.05) is proved to have statistical significance. The gentamicin sulfate has small adverse reaction and better treatment effect on bacillary dysentery. Wangyong et al, discussing the sensitivity of 12 antibiotics to streptococci, demonstrated that gentamicin sulfate is superior in treating streptococci.

In summary, gentamicin sulfate is currently used clinically for the treatment of gram-negative and gram-positive bacterial infections.

Lincomycin hydrochloride injection

Lincomycin hydrochloride is the hydrochloride of lincomycin extracted from the fermented liquid of the variant strain of streptomyces. Lincomycin hydrochloride is a narrow spectrum antibiotic, is mainly effective on gram-positive bacteria, is particularly effective on anaerobic bacteria, pneumococcus and most of streptococcus, and can also be used as a substitute drug for penicillin allergic affected animals.

Lincomycin hydrochloride acts on 50S subunit of sensitive bacterium ribosome, and inhibits the synthesis of mycoprotein by preventing the extension of peptide chain, so as to generate antibacterial action. Lincomycin hydrochloride is generally used as a bacteriostatic agent, but has a bactericidal effect even at a high concentration, and belongs to the 3 rd class of quick-acting bacteriostatic agents in the classification of antibacterial drugs.

Lincomycin hydrochloride is mainly used for treating infection caused by gram-positive bacteria, belongs to a narrow-spectrum antibiotic class, and has the characteristics of less adverse reaction, difficult generation of drug resistance, less drug residue and good pharmacodynamic characteristics. It is mainly effective on gram-positive bacteria such as pneumococcus, streptococcus and anaerobe. The mechanism is that the antibacterial peptide is combined with the central loop of the RNA of the subunit 23r of the 50S of the sensitive bacterium ribosome to prevent the extension of a peptide chain, thereby achieving the aim of inhibiting the synthesis of mycoprotein and producing antibacterial action, which is generally a bacteriostatic agent but also has the bactericidal action at high concentration.

Lincomycin hydrochloride is not easy to absorb by oral administration, is slowly absorbed by intramuscular injection, is widely distributed in body fluid and tissues after being absorbed, can permeate into bone tissues, and has higher concentration in bone marrow. About 50% are metabolized in the liver and the metabolites remain active.

In the existing antibiotics, because part of bacteria have drug resistance, the dosage is often required to be increased, however, the following findings are found:

1. the lincomycin hydrochloride injection has the effect of blocking neuromuscular. Intramuscular administration may cause transient diarrhea or soft stools. Although rare, the necessary measures should be taken to prevent dehydration if it occurs.

2. The main adverse reactions of gentamicin sulfate are damage to kidney and the eighth to cranial nerve, and in addition, the gentamicin sulfate also has respiratory depression effect, and is not suitable for bolus injection. Where ototoxic properties cause vestibular damage that worsens with the accumulation of medication.

Therefore, in order to reasonably use the existing antibiotics, avoid the generation of further drug resistance of bacteria and reduce the dosage, the applicant designs a compound gentamicin sulfate injection.

The design principle of the invention is as follows:

theoretically, the 2 nd type and the 3 rd type antibiotics can be used together to obtain additive or synergistic effect, the lincomycin hydrochloride and the gentamicin sulfate respectively act on 50S and 30S subunits of bacterial ribosome to block the synthesis of the protein through different ways, and the bacterial cell membrane is damaged, so that the rapid bacteriostasis and sterilization effects are achieved.

The combination of the two medicines can quickly inhibit multiple processes of bacterial protein synthesis to sterilize. Andreeva P et al report that gentamycin sulfate and lincomycin hydrochloride in vitro combined application has synergistic effect on part of staphylococcus aureus and streptococcus agalactiae. The gentamicin sulfate and lincomycin hydrochloride are prepared into a compound preparation, so that the occurrence of drug resistance can be reduced, toxic and side effects can be reduced, and the curative effect on the swine streptococcosis is obviously improved.

On the structural level, lincomycin belongs to lincomycin amine antibiotics, which not only have stronger killing effect on primary pathogen-mycoplasma, but also have stronger killing effect on gram-positive bacteria in secondary pathogens, such as streptococcus suis. Meanwhile, gentamicin as aminoglycoside antibiotic has strong killing effect on gram-negative bacteria in secondary pathogens, such as haemophilus parasuis, thereby realizing the purpose of drug combination.

Example 1

Gentamicin sulfate 10%;

5% of lincomycin hydrochloride;

a synergist: 1% of trimethoprim;

antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: 20% of propylene glycol;

The preparation method comprises the following steps: (taking a total of 1000ml as an example)

1. Measuring 60% of total amount of water for injection, adding 2g of sodium metabisulfite and 20g of sodium citrate, and stirring for dissolving;

2. taking propylene glycol (200ml) with the prescription amount, adding 10g of trimethoprim, heating to 50-60 ℃, and stirring for dissolving;

3. mixing the above two solutions, and stirring;

4. respectively adding 100g of gentamicin sulfate and 50g of lincomycin hydrochloride, and stirring for dissolving;

5. adjusting pH to 4.5-6.5 with citric acid;

6. adding water to the total amount, filtering with 0.45um polyethersulfone filter core, introducing nitrogen, bottling, and sterilizing at 121 deg.C for × 15 min.

Comparative example 1

A veterinary compound gentamicin sulfate injection contains the following components in percentage by weight.

Gentamicin sulfate 10%;

5% of lincomycin hydrochloride;

antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

the balance of water for injection, and the pH is adjusted to 4.5-6.5 by citric acid.

The procedure was as in example 1.

Compared with example 1, the comparative example 1 has no synergist, and the same dosage of the synergist is poorer in efficacy than the synergist.

Comparative example 2

Gentamicin sulfate 10%;

5% of lincomycin hydrochloride;

a synergist: 2 percent of trimethoprim

Antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: 30% of propylene glycol;

Compared with example 1, the synergist in this example has double effect, same effect and no double effect compared with example 1.

Comparative example 3

5% of gentamicin sulfate;

10% of lincomycin hydrochloride;

a synergist: 1 percent of trimethoprim

Antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: 20% of propylene glycol;

The preparation procedure was the same as in example 1,

compared with the example 1, the content of the main drug is adjusted, and the effect is poor at the same dosage.

Comparative example 4

Gentamicin sulfate 10%;

10% of lincomycin hydrochloride;

a synergist: 1 percent of trimethoprim

Antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: propylene glycol 20%

The preparation procedure was the same as in example 1,

compared with the embodiment 1, the content of the main drug is adjusted, and the difference of the drug effects of the same dosage is not obvious.

And the main components of the prior patent 201010176388.2 include:

4% of gentamicin sulfate;

2% of ofloxacin;

30% of analgin;

1.5 percent of trimethoprim.

In order to strengthen the standard management of veterinary drugs and ensure the safety, effectiveness, controllable quality and animal food safety of the veterinary drugs, the compound preparation of the antibiotic for injection and the chemical synthetic drugs of analgin and fluoroquinolone is definitely abandoned according to the relevant regulations of the state.

However, the gentamicin sulfate of the prior patent 201010176388.2 belongs to the antibiotics for injection and the ofloxacin belongs to the chemical synthesis drugs of fluoroquinolone, and the formula contains 30% of analgin as antipyretic analgesic. The formulation may pose a hazard or potential risk to the aquaculture industry, human health, or the like.

The invention has the advantages that:

gentamicin is effective against a variety of gram-negative bacteria and staphylococcus aureus. Most streptococcus, anaerobe, etc. are resistant to the medicine.

Lincomycin is mainly resistant to gram-positive bacteria and anaerobic bacteria infection. It is commonly used clinically to treat gram-positive bacterial and mycoplasma infections.

When the gentamicin and the lincomycin are used together, the antibacterial spectrum can be expanded, the drug effect is increased, and the synergistic effect on gram-positive bacteria such as staphylococcus, streptococcus and the like is remarkable. Meanwhile, the compound can be used for mycoplasma infection and the like, and the gentamicin and the lincomycin can be used for food animals.

The trimethoprim is added, so that the drug effect of the compound preparation can be obviously improved, and the curative effect is enhanced.

Firstly, data results of destructive tests and influence factor tests of gentamycin sulfate and lincomycin hydrochloride injection.

The stability tests of the compound preparation (gentamicin sulfate 10%, lincomycin hydrochloride 5%, trimethoprim 1%), comparative example 3 (gentamicin sulfate 5%, lincomycin hydrochloride 10%, trimethoprim 1%), comparative example 4 (gentamicin sulfate 10%, lincomycin hydrochloride 10%, trimethoprim 1%) sample preparation: light test, accelerated test, and long-term test

(1) Light test

The content changes are shown in the following table 1:

TABLE 1

The pH changes are shown in table 2 below:

TABLE 2

The color change is as follows in table 3:

TABLE 3

(2) Accelerated testing of 3 sample formulations

The content changes are shown in the following table 4:

TABLE 4

The pH changes are shown in table 5 below:

the color changes are shown in table 6 below:

TABLE 6

(3) The long-term testing of the 3 sample formulations is shown in table 7 below:

TABLE 7

From the above table, it can be derived:

the temperature stability of 3 formulations was obtained according to accelerated test, long term test: examples 4 and 5 were slightly less stable, while the combination formulation was substantially stable.

According to stability test results, the approach degree of the compound preparation to the physiological adaptive pH value and other factors, the formula of the compound preparation (gentamycin sulfate 10%, lincomycin hydrochloride 5% and trimethoprim 1%) is the optimal formula.

(3) Phase II clinical animal test data

160 sick pigs diagnosed with the streptococcus suis disease are selected and determined from a certain pig farm, and comparative example 3 (gentamycin sulfate 5%, lincomycin hydrochloride 10% and trimethoprim 1%), comparative example 4 (gentamycin sulfate 10%, lincomycin hydrochloride 10% and trimethoprim 1%), single lincomycin and single amoxicillin are selected to be grouped and administered respectively.

The clinical efficacy of the test groups after administration is shown in Table 8:

TABLE 8

As can be seen from the above table 8, the cure rates of the intramuscular injection of the compound preparation to natural case streptococcus suis in the experiment are 83.3% and 81.0% respectively, and the effective rates are 92.7% and 90.3% respectively; the cure rates of the control group of lincomycin and amoxicillin to the natural case streptococcus suis are respectively 75.5%, 70.1%, 66.0% and 65.5%, and the effective rates are respectively 82.6%, 80.2%, 73.0% and 72.8%.

The compound preparation has obvious difference (0.01< P <0.05) between the high and medium dose groups and the control groups of the comparative examples 3 and 4; the difference of the high and medium dosage groups of the compound preparation and the lincomycin and amoxicillin control groups is extremely remarkable (p is less than 0.01) compared with the drug administration group. The high and medium dosage groups of the compound preparation have good curative effect on the streptococcus suis.

Claims

1. The compound gentamicin sulfate injection is characterized by comprising the following components in percentage by weight:

gentamicin sulfate 10%;

5% of lincomycin hydrochloride;

a synergist: 1% of trimethoprim;

antioxidant: 0.2 percent of sodium metabisulfite;

a stabilizer: 2% of sodium citrate;

solvent: 20% of propylene glycol;

2. The preparation method of the compound gentamicin sulfate injection as claimed in claim 1, comprising the following steps:

adjusting the pH to 4.5-6.5 with the citric acid;

3. The use of the compound gentamicin sulfate injection as claimed in claim 1, wherein the use is for preparing a medicament for treating streptococcus suis infection.

4. The use of the compound gentamicin sulfate injection as claimed in claim 3, wherein the treatment of swine streptococcosis comprises treatment of infection of Streptococcus suis or Haemophilus parasuis.