阅读说明：本技术 包含奥硝唑类化合物的药物组合物及其制备方法和用途 (Pharmaceutical composition containing ornidazole compounds and preparation method and application thereof ) 是由 杨丹 陈令武 丁菲 张琪 杨海雪 汤传飞 王嬿钧 梁衡 王旭艳 于 2020-09-28 设计创作，主要内容包括：本发明公开了包含奥硝唑类化合物的药物组合物及其制备方法和用途。该药物组合物包含奥硝唑类化合物和含量在1000mg/kg以下的1-(2,3-环氧丙烷)-2-甲基-5-硝基咪唑。本发明的药物组合物有效期内的安全性和/或稳定性取得显著改善。(The invention discloses a pharmaceutical composition containing ornidazole compounds, a preparation method and application thereof. The pharmaceutical composition comprises ornidazole compounds and 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole with the content of below 1000 mg/kg. The pharmaceutical composition of the present invention has significantly improved safety and/or stability over the useful life.)

1. A pharmaceutical composition is characterized by comprising an ornidazole compound and 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole with the content of below 1000 mg/kg;

the ornidazole compound can be selected from ornidazole, stereoisomer thereof or precursor compound thereof, preferably from ornidazole, levo-ornidazole or dextro-ornidazole or precursor compound thereof;

the ornidazole, l-ornidazole or d-ornidazole precursor compound may be selected from its pharmaceutically acceptable precursor compounds, for example from ornidazole or l-ornidazole ester or pharmaceutically acceptable salt of the ester or their hydrates, for example l-ornidazole phosphate disodium salt or its hydrate.

2. Pharmaceutical composition according to claim 1, characterized in that the content of ornidazole as active ingredient in the pharmaceutical composition may be in the range of 10% to 99%, preferably 15% to 90%, such as 20% to 85% by weight of ornidazole, l-ornidazole disodium phosphate or any hydrate thereof.

3. The pharmaceutical composition according to claim 1 or 2, wherein the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole has a structure represented by the following formula (II):

the content of the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is preferably 100mg/kg or less, more preferably 60mg/kg or less.

4. The pharmaceutical composition of any one of claims 1-3, further optionally comprising or excluding a compound of formula (IV):

；

preferably, in the pharmaceutical composition, the content of the compound represented by the formula (IV) is 15000mg/kg or less;

preferably, the pharmaceutical composition optionally further comprises or does not comprise 2-methyl-5-nitroimidazole;

preferably, in the pharmaceutical composition, the content of the 2-methyl-5-nitroimidazole in percentage by weight is below 2000 mg/kg;

preferably, the pharmaceutical composition further optionally comprises or does not comprise 1- (2, 3-dihydroxypropyl) -2-methyl-5-nitroimidazole represented by the following formula (III):

preferably, in the pharmaceutical composition, the content of the 1- (2, 3-dihydroxypropyl) -2-methyl-5-nitroimidazole is below 2100 mg/kg.

5. The pharmaceutical composition according to any one of claims 1 to 4, further comprising a pharmaceutically acceptable excipient;

preferably, the pharmaceutically acceptable excipients are selected from at least one of the following excipients including but not limited to: fillers, disintegrants, binders, lubricants, surfactants, flavoring agents, wetting agents, pH adjusting agents, solubilizers or cosolvents, and osmotic pressure adjusting agents.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pharmaceutical composition is a formulation, preferably a formulation for gastrointestinal administration or a formulation for parenteral administration;

preferably, the pharmaceutical composition is an injection or a freeze-dried preparation for injection, wherein the pharmaceutical composition comprises an ornidazole compound, preferably ornidazole, levo-ornidazole, disodium levo-ornidazole phosphate or any hydrate thereof, and 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole with the content of below 60 mg/kg;

preferably, the pharmaceutical composition contains citric acid, and the content of the citric acid in percentage by weight is 10-35%;

preferably, the injection has a pH of 4.0 to 6.0;

preferably, the injection solution may contain water for injection, ethanol and/or propylene glycol.

7. The method for preparing the pharmaceutical composition of any one of claims 1-6, wherein the method comprises mixing the ornidazole compound with pharmaceutically acceptable auxiliary materials to obtain the pharmaceutical composition.

Preferably, the l-ornidazole disodium phosphate or the hydrate thereof is prepared into a solution with water, and the solution is freeze-dried to obtain the pharmaceutical composition.

Preferably, the pH of the solution is 4.0-6.0, preferably 4.5-5.5.

Preferably, the temperature of the freeze drying is-60 ℃ to 30 ℃, preferably-50 ℃ to 25 ℃.

Preferably, the freeze-drying time is from 30 to 70h, such as from 40 to 60h, such as from 45 to 55 h.

Preferably, the freeze-drying is temperature programmed.

8. The pharmaceutical composition of any one of claims 1-6 for use in improving pharmaceutical safety.

9. The pharmaceutical composition of any one of claims 1-6 for use in improving drug stability.

10. The pharmaceutical composition of any one of claims 1-6 for use in the preparation of a medicament.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition containing ornidazole compounds, and a preparation method and application thereof.

Background

The ornidazole is a nitroimidazole derivative, is a drug for strongly resisting anaerobic bacteria and protozoan infection, and is a third-generation nitroimidazole derivative which is newly developed after metronidazole, and has higher curative effect, shorter treatment course, better tolerance and wider in-vivo distribution. The antimicrobial action of ornidazole is through the reduction of the nitro group in its molecule to an amino group in an anaerobic environment or through the formation of free radicals interacting with cellular components leading to the death of the microorganism. Ornidazole is 1- (3-chloro-2-hydroxypropyl) -2-methyl-5-nitroimidazole (CAS No.16773-42-5), and has a chemical structure shown in the following formula:

l-ornidazole (also known as "L-ornidazole", CAS No.166734-83-4) is L-isomer of ornidazole, and is mainly used for treating bacteroides fragilis, bacteroides dieselii, bacteroides ovatus, bacteroides thetaiotaomicron, bacteroides vulgatus, clostridium, eubacterium, digestive coccus and digestive streptococcus, helicobacter pylori, bacteroides melanoides, clostridium, CO₂And various infectious diseases caused by anaerobic bacteria such as the bacteria of the weaven fungi and the gingival bacteria, or used for preventing the infection before operation. L-ornidazole has the chemical structure shown in the following formula:

the research shows that compared with dextroisomer of ornidazole or its racemate, levo-ornidazole has lower neurotoxicity and thus obviously improved safety. In addition, prodrugs of l-ornidazole have been developed, including phosphate esters of l-ornidazole (also known as l-ornidazole phosphate) or salts thereof (e.g., disodium l-ornidazole phosphate). After the prodrug is administrated, the prodrug can be rapidly degraded into the levoornidazole under the action of in vivo phospholipase so as to exert the drug effect.

At present, the characteristics of the ornidazole, its stereoisomer and its prodrug, especially the safety and/or stability of the ornidazole, l-ornidazole and their prodrug during the effective period, are still paid extensive attention by drug researchers, and the research direction is needed.

Disclosure of Invention

In order to improve the technical problem, the invention provides a pharmaceutical composition, which comprises an ornidazole compound and 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole with the content of below 1000 mg/kg.

According to an embodiment of the present invention, the ornidazole-based compound may be selected from ornidazole, its stereoisomers or their precursor compounds. As an example, the ornidazole compound may be selected from ornidazole, levo-ornidazole (also known as "levo-ornidazole") or dextro-ornidazole (also known as "dextro-ornidazole") or their precursor compounds.

According to an embodiment of the present invention, the ornidazole has the chemical structure shown in formula (I-1) below:

the L-ornidazole has the structure shown in the following formula (I-2):

the D-ornidazole has the structure shown in the following formula (I-3):

according to an embodiment of the present invention, the ornidazole, l-ornidazole or d-ornidazole precursor compound may be selected from its pharmaceutically acceptable precursor compound, for example from an ester of any of them or a pharmaceutically acceptable salt of the ester, and a hydrate of them, for example, at least one of an amino acid ester, a phosphoric acid ester, an amino acid salt of a phosphoric acid ester, and a salt of a phosphoric acid ester with an alkali metal or alkaline earth metal ion or a hydrate of them; for example, at least one of a sodium salt, a potassium salt, a calcium salt, a magnesium salt, etc. of phosphoric acid ester or a hydrate thereof, and an illustrative example is disodium phosphate salt or a hydrate thereof.

According to an embodiment of the present invention, the precursor compound of l-ornidazole may be selected from at least one of l-ornidazole amino acid ester, l-ornidazole phosphate, l-ornidazole amino acid phosphate, and salts of l-ornidazole phosphate with alkali metal or alkaline earth metal ions or their hydrates; for example, at least one of a sodium salt, a potassium salt, a calcium salt, a magnesium salt, etc. of l-ornidazole phosphate or a hydrate thereof, and exemplified by l-ornidazole phosphate disodium salt (CAS No.909133-95-5) or a hydrate thereof.

According to an embodiment of the present invention, the ornidazole, l-ornidazole or d-ornidazole precursor compound may be present in the pharmaceutical composition in the form of its amorphous or polymorphic forms. Or alternatively, the ornidazole, the precursor compound of l-ornidazole or d-ornidazole may also be selected from solvates of pharmaceutically acceptable salts of esters of any of them, such as hydrates thereof, for example at least one of 1, 2,3, 4, 5, 6 or 7 hydrates of the pharmaceutically acceptable salts of the esters, examples of which may be selected from hydrates of the disodium salt of l-ornidazole phosphate, such as at least one of 5 hydrates, 6 hydrates, 7 hydrates thereof.

According to an embodiment of the present invention, 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole has a structure as shown in the following formula (II):

。

according to an embodiment of the invention, the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is present in the pharmaceutical composition in an amount of less than 900mg/kg, such as less than 800mg/kg, less than 700mg/kg, less than 600mg/kg, less than 500mg/kg, less than 400mg/kg, less than 300mg/kg, less than 200mg/kg, preferably less than 100 mg/kg. Preferably, the content of 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is 90mg/kg or less, for example 80mg/kg or less, 70mg/kg or less, more preferably 60mg/kg or less, 50mg/kg or less, 40mg/kg or less, 30mg/kg or less, 20mg/kg or less or 10mg/kg or less.

According to a preferred embodiment of the invention, the content of 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole in the pharmaceutical composition is below 9mg/kg, such as below 8mg/kg, below 7mg/kg, more preferably below 6mg/kg, examples of which may be below 5mg/kg, below 4mg/kg, below 3mg/kg, below 2mg/kg or below 1 mg/kg.

According to a more preferred embodiment of the invention, the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is contained in the pharmaceutical composition in an amount of 0.9mg/kg or less, such as 0.8mg/kg or less, 0.7mg/kg or less, more preferably 0.6mg/kg or less, and examples thereof may be 0.5mg/kg or less, 0.4mg/kg or less, 0.3mg/kg or less, 0.2mg/kg or less, or 0.1mg/kg or less.

According to an embodiment of the present invention, the content of 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole in the pharmaceutical composition may be 0, which means that 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is not contained in the pharmaceutical composition. In some embodiments of the pharmaceutical compositions of the present invention, the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is present in an amount greater than 0, which means that the pharmaceutical composition contains 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole.

According to an embodiment of the present invention, the content of the ornidazole compound as an active ingredient in the pharmaceutical composition may be in the range of 10% to 99%, preferably 15% to 90%, for example 20% to 85%, and examples thereof may be in the range of, for example, 72% to 83%, and examples thereof may be 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, based on the l-ornidazole, l-ornidazole disodium phosphate or a hydrate of any of them.

According to an embodiment of the present invention, the pharmaceutical composition may optionally further comprise or not comprise a compound represented by the following formula (IV):

。

according to an embodiment of the invention, the content of the compound of formula (IV) in the pharmaceutical composition may be 15000mg/kg or less, such as 14000mg/kg or less, 13000mg/kg or less, 12000mg/kg or less, 11000mg/kg or less, 10000mg/kg or less, 8000mg/kg or less, 7000mg/kg or less, 6000mg/kg or less, 5000mg/kg or less, 4000mg/kg or less, 3000mg/kg or less, 2000mg/kg or less. Preferably, the compound of formula (IV) may be present in an amount of 1000mg/kg or less, for example 900mg/kg or less, 800mg/kg or less, 700mg/kg or less, 600mg/kg or less, 500mg/kg or less, 400mg/kg or less, 300mg/kg or less, 200mg/kg or less or 100mg/kg or less.

According to a preferred embodiment of the invention, the content of the compound of formula (IV) in the pharmaceutical composition is below 90mg/kg, such as below 80mg/kg, below 70mg/kg, more preferably below 60mg/kg, examples of which may be below 50mg/kg, below 40mg/kg, below 30mg/kg, below 20mg/kg, below 10mg/kg or 0. Wherein, when the content of the compound shown in the formula (IV) in the pharmaceutical composition is 0, the pharmaceutical composition does not contain the compound shown in the formula (IV). Alternatively, the content of the compound of formula (IV) in the pharmaceutical composition may be > 0.

According to an embodiment of the present invention, 2-methyl-5-nitroimidazole may also optionally be included or not included in the pharmaceutical composition.

According to an embodiment of the invention, the 2-methyl-5-nitroimidazole is present in the pharmaceutical composition in an amount of less than 2000mg/kg, such as less than 1900mg/kg, less than 1800mg/kg, less than 1700mg/kg, less than 1600mg/kg, less than 1500mg/kg, preferably less than 1400mg/kg, less than 1300mg/kg, less than 1200mg/kg, less than 1100mg/kg, less than 1000mg/kg, such as less than 900mg/kg, less than 800mg/kg, less than 700mg/kg, less than 600mg/kg, less than 500mg/kg, less than 400mg/kg, less than 300mg/kg, less than 200mg/kg or less than 100 mg/kg.

According to a preferred embodiment of the invention, the 2-methyl-5-nitroimidazole content in the pharmaceutical composition is below 90mg/kg, such as below 80mg/kg, below 70mg/kg, more preferably below 60mg/kg, examples of which may be below 50mg/kg, below 40mg/kg, below 30mg/kg, below 20mg/kg, below 10mg/kg or 0. Wherein, when the content of the 2-methyl-5-nitroimidazole in the pharmaceutical composition is 0, the pharmaceutical composition does not contain the 2-methyl-5-nitroimidazole.

According to an exemplary embodiment of the present invention, when the pharmaceutical composition comprises a precursor compound of l-ornidazole, it may or may not comprise l-ornidazole, preferably wherein the l-ornidazole is present in an amount of less than 2000mg/kg, such as less than 80mg/kg, less than 70mg/kg, more preferably less than 60mg/kg, examples of which may be less than 50mg/kg, less than 40mg/kg, less than 30mg/kg, less than 20mg/kg or less than 10mg/kg or 0. Wherein, when the content of the l-ornidazole in the pharmaceutical composition is 0, the pharmaceutical composition does not contain the l-ornidazole.

According to an embodiment of the present invention, the pharmaceutical composition may optionally further comprise or not comprise 1- (2, 3-dihydroxypropyl) -2-methyl-5-nitroimidazole represented by the following formula (III):

according to an embodiment of the invention, the 1- (2, 3-dihydroxypropyl) -2-methyl-5-nitroimidazole is present in the pharmaceutical composition in an amount of 2100mg/kg or less, such as 2000mg/kg or less, 1900mg/kg or less, 1800mg/kg or less, 1700mg/kg or less, 1600mg/kg or less, 1500mg/kg or less, 1400mg/kg or less, 1300mg/kg or less, 1200mg/kg or less, 1100mg/kg or less.

According to an embodiment of the invention, the 1- (2, 3-dihydroxypropyl) -2-methyl-5-nitroimidazole is present in the pharmaceutical composition in an amount of 1000mg/kg or less, for example 900mg/kg or less, 800mg/kg or less, 700mg/kg or less, 600mg/kg or less, 500mg/kg or less, 400mg/kg or less, 300mg/kg or less, 200mg/kg or less or 100mg/kg or less.

According to a preferred embodiment of the invention, the content of the compound of formula (III) in the pharmaceutical composition is below 90mg/kg, such as below 80mg/kg, below 70mg/kg, more preferably below 60mg/kg, examples of which may be below 50mg/kg, below 40mg/kg, below 30mg/kg, below 20mg/kg or below 10 mg/kg.

According to a more preferred embodiment of the invention, the content of the compound of formula (III) in the pharmaceutical composition is below 9mg/kg, such as below 8mg/kg, below 7mg/kg, more preferably below 6mg/kg, examples of which may be below 5mg/kg, below 4mg/kg, below 3mg/kg, below 2mg/kg, below 1mg/kg or 0. Wherein, when the content of the compound shown in the formula (III) in the pharmaceutical composition is 0, the pharmaceutical composition does not contain the compound shown in the formula (III). Alternatively, the content of the compound of formula (III) in the pharmaceutical composition may be > 0.

According to an embodiment of the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable adjuvant, such as a carrier or excipient. The pharmaceutically acceptable excipients are preferably chemically unreactive or inert towards the active ingredient. For example, the pharmaceutically acceptable excipients are selected from at least one of the following, including but not limited to: fillers, disintegrants, binders, lubricants, surfactants, flavoring agents, wetting agents, pH adjusting agents, solubilizers or cosolvents, osmotic pressure adjusting agents, and the like.

According to the technical scheme of the invention, the filler can be at least one selected from lactose, sucrose, glucose, mannitol, sorbitol, calcium sulfate, calcium gluconate, calcium hydrophosphate, calcium phosphate, calcium carbonate, calcium bicarbonate, starch, carboxymethyl starch, pregelatinized starch, microcrystalline cellulose and the like.

According to the technical scheme of the invention, the disintegrating agent can be selected from at least one of pregelatinized starch, microcrystalline cellulose, alginic acid, lignocellulose, sodium carboxymethyl starch, guar gum, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose and the like.

According to the technical scheme of the invention, the adhesive can be at least one selected from gelatin, dextrin, maltodextrin, sucrose, Arabic gum, polyvinylpyrrolidone, methylcellulose, carboxymethylcellulose, ethyl cellulose, polyvinyl alcohol, polyethylene glycol, hydroxypropyl methylcellulose and the like.

According to the technical scheme of the invention, the lubricant can be selected from at least one of magnesium stearate, calcium stearate, zinc stearate, talcum powder, glyceryl monostearate, polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 8000, sodium benzoate, adipic acid, fumaric acid, boric acid, sodium chloride, sodium oleate, glyceryl triacetate, polyoxyethylene glyceryl monostearate, monolaurocysteine, sodium chloride, sodium lauryl sulfate, magnesium lauryl sulfate and the like.

According to the technical scheme of the invention, the surfactant can be selected from at least one of sodium dodecyl sulfate, poloxamer, polysorbate 80, hexadecyl trimethylamine bromide, sodium lauryl sulfate, sodium stearate, polyoxyethylene higher fatty alcohol, sucrose ester, sorbitol fatty ester, soybean phospholipid and the like.

According to the technical scheme of the invention, the flavoring agent is at least one of steviosin, fructose, glucose, high fructose syrup, honey, aspartame, xylitol, mannitol, lactose, sorbitol, essence, maltitol and the like.

According to the technical scheme of the invention, the pH regulator can be at least one selected from hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, triethylamine and the like.

According to the technical scheme of the invention, the cosolvent can be selected from alcohol solvents, for example, at least one selected from ethanol, glycerol, propylene glycol, polyethylene glycol (such as polyethylene glycol 300, polyethylene glycol 400, and the like) and the like.

According to the technical scheme of the invention, the osmotic pressure regulator can be at least one selected from sodium chloride, glucose, fructose, phosphate, polyethylene glycol, propylene glycol, mannitol and the like.

According to the technical scheme of the invention, the pharmaceutical composition can be a preparation, for example, a gastrointestinal administration preparation or a parenteral administration preparation. Wherein the gastrointestinal tract administration preparation can be tablets, dispersible tablets, capsules, sustained-release agents, granules, oral liquid, syrup and the like; the parenteral administration preparation can be an infusion preparation, an injection (such as a liquid injection), a freeze-dried preparation (such as freeze-dried powder), an effervescent tablet, a suppository, a sublingual tablet and the like, and is preferably a capsule, an injection (such as a liquid injection) or a freeze-dried preparation (such as freeze-dried powder) for injection.

According to an exemplary embodiment of the present invention, the pharmaceutical composition is an injection or a lyophilized preparation for injection, which comprises an ornidazole compound, preferably ornidazole, disodium levoornidazole phosphate or a hydrate of any of them, and 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole in an amount of less than 60 mg/kg.

According to an embodiment of the present invention, the content of the ornidazole compound as an active ingredient in the injection or lyophilized preparation for injection is in the range of 70% to 85% by weight, for example, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 82%, 83%, 84%, 85%, preferably in the range of 72% to 83% of ornidazole or its isomer.

According to an embodiment of the invention, the pharmaceutical composition, e.g. a lyophilized formulation for injection, may contain citric acid in an amount of 10-35% by weight, preferably 15-32%, e.g. 18-28%, 20-25%, exemplarily 16%, 17%, 18%, 19%, 21%, 22%, 23%, 24%, 26%, 27%, 29%.

According to an embodiment of the invention, the pH of the injection may be 4.0-6.0, preferably 4.5-5.5, e.g. 4.6-5.4, 4.7-5.3, exemplarily 4.8, 4.9, 5.0, 5.1, 5.2.

According to the embodiment of the invention, when the pH of the injection is 4.0-6.0, the content of citric acid can be 19-45 mg/mL. For example, at pH 4.0, the citric acid content is 45 mg/mL; when the pH value is 6.0, the content of the citric acid is 19 mg/mL.

According to an embodiment of the present invention, the injection solution may include water for injection, ethanol and/or propylene glycol.

According to the technical scheme of the invention, the administration route of the pharmaceutical composition comprises but is not limited to gastrointestinal administration or parenteral administration; the parenteral administration may be injection (e.g., intravenous injection, arterial injection, intramuscular injection, subcutaneous injection, intradermal injection, etc.), vaginal administration, mucosal administration, or the like.

According to an embodiment of the present invention, the active ingredient of the pharmaceutical composition is an ornidazole compound, preferably ornidazole, levo-ornidazole or a precursor compound of levo-ornidazole. Preferably, the pharmaceutical composition may not contain other active ingredients than the ornidazole-type compound, or may also contain other active ingredients than ornidazole, levoornidazole or a precursor compound of levoornidazole.

According to an exemplary embodiment of the invention, the pharmaceutical composition comprises either l-ornidazole disodium phosphate and citric acid, and optionally water for injection, either present or absent. Preferably, the pH of the pharmaceutical composition is 4.5-5.5.

According to an exemplary embodiment of the invention, the pharmaceutical composition comprises ornidazole, ethanol, propylene glycol and optionally water for injection, either present or absent.

The invention also provides a preparation method of the pharmaceutical composition, which comprises the step of mixing the ornidazole compound with pharmaceutically acceptable auxiliary materials to obtain the pharmaceutical composition.

According to an exemplary embodiment of the present invention, ornidazole, ethanol, propylene glycol and water for injection are mixed to obtain the pharmaceutical composition.

According to an exemplary embodiment of the present invention, the disodium l-ornidazole phosphate or its hydrate is prepared into a solution with water, and the solution is freeze-dried to obtain the pharmaceutical composition.

According to an embodiment of the invention, the pH of the solution is 4.0-6.0, preferably 4.5-5.5, such as 4.6-5.4, 4.7-5.3, exemplarily 4.8, 4.9, 5.0, 5.1, 5.2.

According to an embodiment of the invention, the temperature of the freeze-drying is between-60 ℃ and 30 ℃, preferably between-50 ℃ and 25 ℃.

According to an embodiment of the invention, the freeze-drying time is 30-70h, such as 40-60h, further such as 45-55 h.

According to an embodiment of the invention, the freeze-drying is temperature programmed.

According to an aspect of the present invention, an exemplary operation of the freeze-drying includes: sending the semi-finished product filled with the semi-stopper into a freeze-drying box when the temperature of the silicone oil is reduced to 0 ℃, keeping for 0.5 hour, and keeping for 1.0 hour when the temperature of the silicone oil is reduced to-50 ℃; heating to-20 deg.C, and maintaining for 3.0 h; cooling to-50 deg.C, and maintaining for 3.0 h. The vacuum pump is opened, the vacuum is pumped to 200 mu bar, the temperature is raised to-20 ℃ within 1.0h, and the temperature is maintained for 16.0 h. Heating to-15 deg.C for 5min, and maintaining for 6.0 h; heating to-10 deg.C for 5min, and maintaining for 1.0 h; heating to 0 deg.C for 10min, and maintaining for 1.0 h; heating to 10 deg.C for 10min, and maintaining for 1.0 h; heating to 25 deg.C for 15min, and maintaining for 7.0 h; extreme vacuum was applied at 25 ℃ and continued for 13.0 h. Performing pressure rise test (the pressure rise qualification standard is less than or equal to 15 μ bar/min), freeze-drying after qualification, filling nitrogen into the box body to break vacuum (vacuum control range: 850mbar-900mbar), and taking out of the box after full pressure plugging.

The invention also provides the use of the pharmaceutical composition for improving the safety of a medicament.

The invention also provides the use of the pharmaceutical composition for improving the stability of a medicament.

The invention also provides the application of the pharmaceutical composition in preparing medicines.

Preferably, the medicament is used for preventing or treating diseases related to anaerobes, such as infectious diseases caused by anaerobes. For example, the anaerobic bacteria may be selected from at least one of the following, including but not limited to: bacteroides fragilis, Bacteroides dirichi, Bacteroides ovatus, Bacteroides thetaiotaomicron, Bacteroides vulgatus, Clostridium clostridia, Eubacterium, Pediococcus and Pediococcus, helicobacter pylori, Bacteroides melanoides, Fusobacterium, CO₂And anaerobic bacteria such as bacteriophage phagemid and gingival bacteroides.

Preferably, the pharmaceutical product is used for preventing or treating infectious diseases caused by anaerobic bacteria before and/or after surgery.

The present invention also provides a method for preventing or treating diseases associated with anaerobes, such as infectious diseases caused by anaerobes, comprising administering a therapeutically effective amount of the pharmaceutical composition to a patient in need thereof.

Definition and description of terms

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present specification.

The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound of the present invention sufficient to effect the intended use, including but not limited to the treatment of a disease as defined below. The therapeutically effective amount may vary depending on the following factors: the intended application (in vitro or in vivo), or the subject and disease condition being treated, such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc., can be readily determined by one of ordinary skill in the art. The specific dosage will vary depending on the following factors: the particular compound selected, the dosage regimen to be followed, whether to administer it in combination with other compounds, the timing of administration, the tissue to be administered and the physical delivery system carried.

The term "patient" refers to a patient in need of a targeted prevention or treatment of a disease associated with anaerobes, wherein the patient is a mammal, e.g., selected from rodents, cows, pigs, dogs, cats and primates, particularly humans.

The term "expiration date" means a period of time during which the quality of a pharmaceutical product is able to meet regulatory requirements under defined storage conditions, for example 24 months, such as 18 months or 12 months.

Advantageous effects

The present application surprisingly found that 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole present in pharmaceutical compositions comprising ornidazole compounds is genotoxic, causes cellular DNA damage, and produces mutagenic effects, creating a great challenge to the safety and/or stability of such drugs during the lifetime of use. Such dosing risks are controlled or even eliminated in the pharmaceutical compositions of the present invention. It was found that the safety and/or stability of such a drug can be significantly improved when the weight percentage content of 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole is reduced to below 1000mg/kg, preferably below 100mg/kg, more preferably below 60 mg/kg.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

Example 1: genotoxicity test of 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole (Compound of formula (II))

Ames experiment (Strain experiment)

1. Materials and methods

1.1 test article

Name: 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole;

packaging: an ampoule bottle;

specification: about 50 mg/count;

storage conditions are as follows: sealing at 2-8 deg.C in dark.

1.2 vehicle information

Name: sodium chloride injection;

production unit: chenxin pharmaceutical industry, gmbh;

properties and physicochemical properties: colorless clear liquid with slightly tasted;

specification: 500 mL/bottle;

storage conditions are as follows: and (5) sealing and storing.

1.3 preparation of test sample/reference

The test sample is prepared on the day of administration, and an appropriate amount of the test sample is weighed and dissolved by adding an appropriate amount of sodium chloride injection to make the concentration of the test sample be 50 mg/mL. The solution was filtered through a 0.2 μm filter and diluted with sodium chloride injection to a concentration of 20, 8, 2, 0.5mg/mL, respectively.

The formulated solution was stored at room temperature prior to administration.

1.4 Experimental methods

The strain is as follows: salmonella typhimurium, histidine-deficient strains (TA97a, TA98, TA100, TA102, TA 1535). The strain is provided by Moltox, and the bacterial liquid is stored in liquid nitrogen.

SD rats are offered by Zhao Zhi (Suzhou) New drug research center, Inc.

Preparing a mixed solution of S9:

SD rat liver S9 fraction with protein concentration of 21.64mg/mL was used in this experiment and stored in liquid nitrogen. Prior to use, the S9 mixture was prepared under sterile conditions, and the S9 mixture was prepared as described in Table 1.

TABLE 1

And (3) enrichment culture of the strain: after the bacterial liquid frozen and preserved by liquid nitrogen is melted in water bath at 37 ℃, 100 mu l of the bacterial liquid is inoculated into 20ml of nutrient broth and cultured for 10 to 12 hours at 37 ℃ in a dark oscillation (120 rpm).

Grouping administration: each test point processed 2 parallel dishes. The administration information is shown in table 2.

TABLE 2

Note: group 1 was a spontaneous control group; groups 2-6 are test article groups.

Taking a corresponding number of glass test tubes, subpackaging 2ml of top layer culture medium (heated at 45-47 ℃) in the test tubes, and then sequentially adding 0.1ml of bacterial culture solution, 0.1ml of test solution, 0.5ml of S9 mixture or PBS with pH 7.4. Quickly and uniformly mixing the mixture on a vibrator, pouring the mixture on the surface of the basic culture medium, and slightly rotating to uniformly spread the mixture on the surface of the basic culture medium.

Placing the plate on a horizontal desktop, inverting the plate after the culture medium is solidified, and culturing at 37 ℃ for 48-72 hours.

1.5 data acquisition

And after culturing for about 48-72 hours, counting the number of the reverted mutant colonies in all the plates, and observing the background lawn under a microscope to evaluate whether the test article has an antibacterial effect on the strains.

The precipitation was observed when the drug was added.

1.6 determination of results

Judging whether the test article has bacteriostatic toxicity to the strain according to the following criteria:

1) the background lawn becomes thinner, and the reduction of the number of revertant colonies can be accompanied;

2) the background lawn disappears, i.e. the bacterial growth is completely inhibited;

3) needle-like non-reverting mutant microcolonies (usually accompanied by background lawn deletion) appear.

2. Results

2.1 precipitation conditions

No precipitate was observed in the sample loading process for each test article dose group.

2.2 bacterial virulence

Background lawn individual data are shown in table 3 below.

Table 3: background lawn individual observation data

Note: "√" indicates that the background lawn is normal; "-" indicates disappearance of background lawn, and X indicates decrease of background lawn.

The results show that: under the condition of non-metabolic activation, under the dosage of 800-5000 mu g/dish, background lawn of each strain disappears, and under the dosage of 50-200 mu g/dish, background lawn of each strain decreases. Under the condition of metabolic activation, under the dosage of 200-5000 mug/dish, background lawn of each strain disappears, and under the dosage of 50 mug/dish, background lawn of each strain is reduced.

2.3 mutagenicity

The results of the counter-mutant colony counts are shown in Table 4 below.

Table 4: return mutation colony count individual data (individual/dish)

The results show that: under the conditions of metabolic activation and non-metabolic activation, the number of the reversion mutation colonies of each strain of the spontaneous control group is within a normal reference range or slightly increased or decreased; under the dosage of 50 and 200 mug/dish in the non-metabolic activation state and the dosage of 50 mug/dish in the metabolic activation state, the number of the reversion mutation colonies of each strain is obviously increased and is 2 times higher than that of a spontaneous control group.

3. Conclusion

Under the test condition, the 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole has a bacterial toxicity effect under the dosage of 50-5000 mu g/dish, has mutagenicity on salmonella typhimurium, and has genotoxicity.

(II) micronucleus test

1. Test and reference substances

1.1 test article

Name: 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole;

packaging: an ampoule bottle;

specification: about 50 mg/count;

storage conditions are as follows: sealing at 2-8 deg.C in dark.

1.2 reference substance

Cyclophosphamide for injection;

production unit: jiangsu Hengrui pharmaceuticals, Inc.;

condition and physicochemical properties: a white crystalline powder;

batch number: 19061521, respectively;

specification: 0.2 g/bottle;

storage conditions are as follows: and (5) shading, sealing and storing.

1.3 Experimental animals

Mouse KM

Grade: SPF;

selling units: henan Spanish Biotech GmbH;

the unit of mass detection: experimental animal center in Shandong province;

license number: SCXK 2020-.

2. Preparation of drug delivery preparation

The preparation process is finished under the aseptic condition, the used reagent bottles and the like are sterilized, and the preparation process is not protected from light.

Solvent (0.9% sodium chloride injection):

production unit: anhui double crane pharmaceutical industry, Inc.;

properties and physicochemical properties: colorless clear liquid, slightly salty taste;

batch number: 20011112C

Specification: 500 mL/bottle;

storage conditions are as follows: and (5) sealing and storing.

Preparation of positive control solution (cyclophosphamide):

weighing the required amount of the positive control (calculated by content), adding a proper amount of solvent, stirring until the positive control is clear and transparent, and finally diluting the positive control to the required concentration by using the solvent. After the preparation is finished, a filter membrane with the diameter of 0.22 mu m is adopted for filtration to obtain a positive control solution.

Preparing a test article administration preparation:

weighing the required amount of the test sample (calculated by content), adding a proper amount of solvent, stirring until the test sample is clear and transparent, and finally diluting the test sample to the required concentration by using the solvent. Filtering with 0.22 μm filter membrane to obtain the test drug delivery preparation.

The doses and concentrations for each group are shown in the following table:

3. experimental system

3.1 Experimental animals

Animals used for the experiment: kunming mouse, clean grade; 24 females and 24 males were selected.

Weight and age at group: female: about 24-28 g, 8-10 weeks old; male: about 25-29 g, 8-10 weeks old.

3.2 grouping

The test has 5 groups, 10 solvent groups and test sample groups, 8 positive control groups and 48 positive control groups.

3.3 animal identification

Animals were identified on the day of reception with 3% picric acid ethanol solution, and grouped together with picric acid and house label as follows:

x is female mouse, Y is male mouse.

3.4 animal randomization group

The animals were randomly grouped according to sex and weight, and the 48 animals selected were assigned to 5 groups. The differences in body weight of the animals used for the grouping were within ± 20% of the mean body weight of the same sex category. After grouping, the average body weight of each group of animals was not statistically different at the 5.0% test level. And (5) storing and recording the rest animals after grouping, and feeding.

3.5 reagent dosage design

The experimental dose design is shown in the following table:

4. test method

4.1 administration of drugs

The administration route is as follows: the intraperitoneal injection administration route is selected.

The administration mode comprises the following steps: the administration is performed 5 times for 3 consecutive days, 1 time in the morning on the first day, and 1 time in the afternoon on the second and third days.

The administration period is as follows: and 3 days.

Dose capacity: 10 mL/kg.

Dosage: the dose was calculated from the last weight weighing.

4.2 Observation and inspection

4.2.1 death or dying

All surviving animals were observed 1 time per day.

4.2.2 body weight

An adaptation period: all animals were weighed 1 time before grouping, i.e. group weight

The administration period is as follows: the body weights were weighed in the morning before D1, D2 and D3 administration, respectively, and used to calculate the dose

4.3 test end-of-line procedure

4.3.1 unintended death animal

During the experiment, all animals did not die or moribund (see section 5.1 for a summary of die or moribund results).

4.3.2 dissection

Planned dissection date: administration end (D4)

And (3) dissecting animals: all surviving animals

The tabletting method comprises the following steps: the mice were sacrificed by cervical dislocation, femurs were taken, muscles were removed, epiphyses were cut off, marrow cavities were exposed, marrow smears were taken, fixed with methanol, and stained with gimesas.

4.3.3 microscopic examination

Planned microscopic examination date: administration end (D5)

The microscopic examination object: all animal bone marrow smears (see details 4.3.2)

The microscopic examination method comprises the following steps: the area with complete cells, uniform dispersion and proper coloring is selected under a low power microscope, 1000 micronucleoli rates of cytoplasmally clear and complete polyphilic red blood cells (PCE) are counted in each smear by observing through a high power microscope, and the result is expressed in per thousand percent (‰).

4.4 statistical analysis

SPSS16.0 statistical software is used for analysis, and the microkernel rate is equal toShow, compare with negative control, using two-sided t-test and chi²The test is statistically processed, P<0.05 is statistically significant, P>0.05 is not significant, 0.01<P<0.05 is significant, P<The difference at 0.01 is very significant.

The death or moribund results were not counted and are expressed as frequency.

5 results

5.1 death or dying

The effect of the test article on the death or moribund outcome of the animals is shown in the following table. By the end of the dosing period, no death or moribund was found in all animals.

5.2 body weight

The results of the body weight examination are shown in the following table.

During the experiment, the animals in the test article administration group showed a weight loss tendency.

The weight of the animals in the 100mg/kg dose group D3 showed a decrease trend and showed dose correlation, and the other animals showed a smaller weight increase and showed dose correlation, although no statistical difference in weight change, and were considered to be related to the test sample.

5.3 Micronuclear Rate of pleochromophilic erythrocytes

The statistic results of the micronucleus rate of the pleochromophilic erythrocytes are shown in the following table.

During the experiment, the animals of the test article administration group showed a rising trend of micronucleus rate.

The micronucleus rates of the 48mg/kg dose group and the 100mg/kg dose group are very different from those of the negative control group (p is less than 0.01), and although no statistical difference is found in other dose groups, the micronucleus rates are also seen to be slightly increased, so that dose correlation is shown, and the micronucleus rates are considered to be related to the test sample.

Conclusion 6

Under the test condition, the Kunming mouse is subjected to intraperitoneal injection for 3 consecutive days to give test compound with the concentration of 2.88, 48 and 100mg/kg, no animal death or dying is seen in the test period, the weight of animals in a test administration group is reduced under the dosage of more than or equal to 100mg/kg, and the rate of the chromophilous erythrocyte micronucleus is increased under the dosage of more than or equal to 48mg/kg, so that the test compound is considered to be related to the test. The level of no observed adverse effect in this test (NOOBSERVED ADDVERSE EFFECT level, NOAEL) was 2.88 mg/kg.

According to the ICH M7 (international human drug registration technical co-ordination) guidelines, genotoxic impurity limits were calculated as follows:

the impurity limit is the acceptable daily intake of the impurity (i.e., PDE value)/maximum daily dosage of the drug.

PDE (NOEL) × body weight/(F1 × F2 × F3 × F4 × F5)

Wherein, the NOEL of the mouse is 2.88 mg/kg/day; f1-12, the human dose was extrapolated from the mouse dose; f2 ═ 10, see FDA ICH-Q3C guidelines; f3 is 10, and the experimental period does not exceed 10 days; f4 ═ 1, no serious toxic reaction was considered to be found; f5 ═ 1; regulating body weight to 50 kg; the calculation results are as follows:

PDE＝2.88mg/kg(NOEL)×50/(12×10×10×1×1)＝0.12mg；

the impurity limit is 0.12mg/2g (the maximum daily dose of the compound in a human body is determined to be 2g according to clinical tests and the like), 60ppm is 60 mg/kg.

Example 2: genotoxicity test of the Compound of formula (IV)

The method of example 1 was used except that 1- (2, 3-epoxypropane) -2-methyl-5-nitroimidazole of formula (II) was replaced with the compound of formula (IV), which showed no genotoxicity at the doses of 50, 200, 800, 2000 and 5000 μ g/dish.

EXAMPLE 3 lyophilized preparation for injection

The formulation of the lyophilized formulation for injection is as follows:

6000g of disodium L-ornidazole phosphate

2700g of citric acid 1140-

Adding water for injection to 60L

30000 pieces of the extract were prepared.

The preparation method of the prescription comprises the following steps:

adding a part of citric acid into injection water with the temperature below 20 ℃ and the total volume of the prescription of 80%, stirring until the citric acid is completely dissolved, adding the levo-ornidazole disodium phosphate according to the prescription amount, stirring until the levo-ornidazole disodium phosphate is completely dissolved, and adjusting the pH value of a preparation solution to 4.0-6.0. And then adding cooled injection water into the residual citric acid to prepare a solution with the concentration of 20g/100mL to adjust the pH of the prepared solution to 4.5-5.5. Adding water for injection (below 20 deg.C) according to the density of the medicinal liquid (1.063g/ml) to the total amount of the prescription. Stirring for about 15min, and filtering the prepared medicinal liquid with filters (0.2 μm/0.22 μm; nylon 66) of A05NF2PH4 and KA3NFP 1.

Sending the filtered liquid medicine into a freeze-drying box when the temperature of the silicone oil is reduced to 0 ℃, keeping for 0.5 hour, and keeping for 1.0 hour when the temperature of the silicone oil is reduced to-50 ℃; heating to-20 deg.C, and maintaining for 3.0 h; cooling to-50 deg.C, and maintaining for 3.0 h. The vacuum pump is opened, the vacuum is pumped to 200 mu bar, the temperature is raised to-20 ℃ within 1.0h, and the temperature is maintained for 16.0 h. Heating to-15 deg.C for 5min, and maintaining for 6.0 h; heating to-10 deg.C for 5min, and maintaining for 1.0 h; heating to 0 deg.C for 10min, and maintaining for 1.0 h; heating to 10 deg.C for 10min, and maintaining for 1.0 h; heating to 25 deg.C for 15min, and maintaining for 7.0 h; extreme vacuum was applied at 25 ℃ and continued for 13.0 h. Performing a pressure rise test (the pressure rise qualification standard is less than or equal to 15 μ bar/min), after the freeze-drying is finished, filling nitrogen into the box body to break vacuum (the vacuum control range is 850mbar-900mbar), and obtaining the freeze-dried preparation.

The above preparation method is repeated, and 3 batches of finished products are obtained by co-production and are marked as S1, S2 and S3, and the immediate test results are shown in Table 5.

TABLE 5 inspection results of the finished products

The accelerated stability of the S1, S2 and S3 products was tested (test conditions: temperature 25 ℃. + -. 2 ℃ C., humidity 60% RH. + -. 5% RH), and the test results are shown in tables 6-8.

TABLE 6S 1 test results for accelerated stability of finished products

TABLE 7S 2 test results for accelerated stability of finished product

TABLE 8S 3 test results for accelerated stability of finished product

The long-term stability of the finished products S1, S2 and S3 was tested (long-term stability test condition: 5 ℃. + -. 3 ℃), and the test results are shown in tables 9-11.

TABLE 9S 1 test results for Long-term stability of finished product

TABLE 10S 2 Long term stability test results for finished products

TABLE 11S 3 Long term stability test results for finished products

The experimental result shows that the freeze-dried preparation has good medication safety and stability.

The detection method of each substance was as follows:

the instrument comprises the following steps: liquid chromatography tandem mass spectrometer, model 1290LC-6470LC/TQ, Agilent technologies, Inc.

(1) Chromatographic conditions

A chromatographic column: svea Core Shell C18 (100 mm. times.4.6 mm. times.2.6 μm)

Mobile phase: a (10mmol ammonium formate aqueous solution) B (methanol)

Flow rate: 0.6mL/min

Sample introduction volume: 5 μ L

Gradient program:

time (min)	Mobile phase a (% by volume)	Mobile phase B (% by volume)
			0.00	90	10
0.20	90	10
			1.00	80	20
2.50	80	20
			4.00	70	30
5.50	70	30
			6.00	10	90
8.00	10	90
			8.01	90	10
10.00	90	10

(2) Conditions of Mass Spectrometry

Type of ion source: AJS ESI ion source

Collision gas (N)₂) Flow rate: 8L/min

Sheath gas (N)₂) Flow rate: 11L/min

Temperature of sheath gas: 350 deg.C

Spraying voltage: 3500V

Nozzle voltage: 500V

EMV gain: 200V

The collection mode is as follows: multiple reactive ion monitoring (MRM).

(3) Preparation of blank solution

Blank solution: i.e. pure water.

(4) Control solution: precisely weighing 10mg of 2-methyl-5-nitroimidazole, the compound of formula (II) and the compound of formula (III) as reference substances, respectively placing the reference substances in a10 mL volumetric flask, and fixing the volume to the scale with blank solution to obtain reference substance stock solution. From these, 0.1mL of the stock solutions of the control samples were precisely measured in 10mL volumetric flasks, and the blank solution was added to the volume to the scale as an intermediate solution of the control sample (i.e., 10. mu.g/mL). From the medium, 5mL of the solution was precisely measured in a 50mL volumetric flask, and the volume was adjusted to the scale with a blank solution as a control working solution (i.e., 1. mu.g/mL). Precisely measuring 0.6mL from the medium into a10 mL volumetric flask, and fixing the volume to the scale with the blank solution to obtain a reference substance solution of 60 ng/mL.

(5) System applicability solution: precisely weighing 10mg (calculated by levo ornidazole phosphate disodium) of a test sample in a10 mL volumetric flask, and fixing the volume of a blank solution to a scale. Taking 1mL of the solution into a10 mL volumetric flask, adding a compound (60mg/kg) of the formula (II), a compound (60mg/kg) of the formula (III) and a reference substance of 2-methyl-5-nitroimidazole (60mg/kg) with the limit concentration (taking 0.6mL of the reference substance working solution under the item (4) into the volumetric flask), and diluting the blank solution to a constant volume (namely 60 ng/mL).

(6) Preparing a test solution: precisely weighing 10mg (calculated by L-ornidazole phosphate disodium) of a sample in a10 mL volumetric flask, and fixing the volume to a scale by using a blank solution to obtain the product.

(7) Preparing a mixed test solution: precisely weighing 10mg of a test sample (calculated by levo ornidazole phosphate disodium) into a10 mL volumetric flask, adding a compound (60mg/kg) with a limit concentration (0.6 mL of a reference working solution under the item of 3.4) in the formula (II), a compound (60mg/kg) in the formula (III) and a reference substance of 2-methyl-5-nitroimidazole (60mg/kg), and fixing the volume of a blank solution to a scale (namely 60 ng/mL).

(8) Limitation: the residual quantity of the compound of formula (II), the compound of formula (III) and the 2-methyl-5-nitroimidazole reference substance in the product is not more than 60 mg/kg.

In the above experimental results, the detection limit concentration of each substance is 0.1ng/mL, and the content Relative Standard Deviation (RSD) meets the acceptable standard.

EXAMPLE 4 lyophilized preparation for injection

Prescription of lyophilized preparation for single injection:

l-ornidazole disodium phosphate 200mg

Citric acid 38-90mg

Adding water for injection to 2mL

The pH value is 4.5-5.5.

Referring to the lyophilization preparation process and detection method in example 3, 12 batches of lyophilized formulations were prepared.

(1) The content of each substance in the lyophilized preparation was measured in real time, and the measurement results are shown in table 12.

TABLE 12 results of sample examination

In the above experimental results, the detection limit concentration of each substance is 0.1ng/mL, and the content Relative Standard Deviation (RSD) meets the acceptable standard.

The other indexes of the 12 batches of freeze-dried preparations all meet the requirements of finished products shown in the table 5.

Example 5 genotoxicity test

The samples of examples 3-4 were tested according to the genotoxicity test method of example 1, and the results showed that: examples 3-4 samples of lyophilized formulations were not genotoxic.

EXAMPLE 6 Ornidazole injection

The prescription of the ornidazole injection is as follows:

ornidazole 500mg

0.9g of ethanol

Propylene glycol 1mL

Water for injection was added to 3 mL.

The preparation process of the ornidazole injection comprises the following steps: mixing the ornidazole, the ethanol, the propylene glycol and the water for injection according to the above dosage to prepare the ornidazole injection. According to the substance detection method in the embodiment 3, the content of the compound of the formula (II) in the ornidazole injection is less than 60 mg/kg.

The samples of example 6 were tested according to the genotoxicity test method of example 1 and the results showed that: the injection of example 6 has no genotoxicity.

The above description has been made of exemplary embodiments of the present invention. However, the scope of the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement and the like made by those skilled in the art within the spirit and principle of the present invention shall be included in the protection scope of the present invention.