CN112778364A - Nitroimidazole derivative and preparation method and application thereof - Google Patents

Nitroimidazole derivative and preparation method and application thereof Download PDF

Info

Publication number
CN112778364A
CN112778364A CN201911075152.7A CN201911075152A CN112778364A CN 112778364 A CN112778364 A CN 112778364A CN 201911075152 A CN201911075152 A CN 201911075152A CN 112778364 A CN112778364 A CN 112778364A
Authority
CN
China
Prior art keywords
compound
nitroimidazole
formula
hours
preparation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201911075152.7A
Other languages
Chinese (zh)
Other versions
CN112778364B (en
Inventor
杨成
刘晓鹏
张起愿
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Huachuang Synthetic Pharmaceutical Co ltd
Original Assignee
Huachuang Synthetic Pharmaceutical Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Huachuang Synthetic Pharmaceutical Co ltd filed Critical Huachuang Synthetic Pharmaceutical Co ltd
Priority to CN201911075152.7A priority Critical patent/CN112778364B/en
Publication of CN112778364A publication Critical patent/CN112778364A/en
Application granted granted Critical
Publication of CN112778364B publication Critical patent/CN112778364B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/645—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having two nitrogen atoms as the only ring hetero atoms
    • C07F9/6503—Five-membered rings
    • C07F9/6506—Five-membered rings having the nitrogen atoms in positions 1 and 3
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention provides a nitroimidazole derivative shown as a general formula I or pharmaceutically acceptable salts, solvent compounds, hydrates, polymorphs, deuterons and isomers thereof; the invention also provides a pharmaceutical composition containing the same and application of the pharmaceutical composition. The nitroimidazole derivative provided by the invention has excellent pharmacokinetic properties and also has excellent antibacterial activity.

Description

Nitroimidazole derivative and preparation method and application thereof
The technical field is as follows:
the invention relates to nitroimidazole derivatives, a preparation method and application thereof, and further relates to nitroimidazole derivatives for resisting anaerobic bacteria and protozoan infection, and a preparation method and application thereof.
Technical background:
the ornidazole is a nitroimidazole derivative, is a strong anti-anaerobe and antiprotozoal infection drug, and is a third-generation nitroimidazole derivative which is newly developed after metronidazole, and has the advantages of 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 formula to an amino group in an anaerobic environment or through the interaction of free radicals with cellular components, leading to the death of the microorganism.
The l-ornidazole is an l-isomer of ornidazole, and Chinese patents CN200510068478.9 and CN200510083517.2 describe that the water solubility is very poor, the melting point is low, the acidity is required to be strong when the injection is prepared into an infusion preparation, and phlebitis is easy to generate when a patient uses the injection; and the toxic degradation product 2-methyl-5 nitroimidazole is higher during high-temperature sterilization of transfusion, thus bringing great insecurity to clinical medication.
The prior art discloses levoornidazole phosphate and medicinal salts thereof, which greatly increase water solubility, but generate impurities in the process of placement, the degradation degree of the levoornidazole phosphate and the medicinal salts thereof is increased along with the increase of temperature, and the toxicity of the degraded impurities is unknown, so that the levoornidazole phosphate and the medicinal salts thereof greatly influence the clinical use.
The compound of the invention also has excellent solubility in water, and the inventor surprisingly finds that the compound of the invention has better stability and does not generate degradation impurities in the process of accelerating placement, and the inventor unexpectedly finds that the pharmacokinetic data of the compound of the invention are superior to that of the disclosed levoornidazole prodrug related compound and have better safety performance in animal experiments.
The invention content is as follows:
the invention aims to provide a novel nitroimidazole derivative which has the following structural formula I:
Figure RE-GDA0002384363250000011
wherein: n is 0 to 12
X is water, methanol, ethanol, acetone, ethyl acetate, isopropanol
R represents hydrogen, alkali metal, meglumine, choline and amino acid.
The object of the present invention is a process for the preparation of the above-mentioned compounds of formula I, which comprises:
(a) reacting a compound shown in a formula A with a compound shown in a formula B in a solvent to form a compound shown in a formula C;
Figure RE-GDA0002384363250000021
(b) reacting the compound shown in the formula C formed in the step (a) with the compound shown in the formula D in an organic solvent in the presence of inert gas to obtain the compound shown in the formula I.
Figure RE-GDA0002384363250000022
The invention aims to provide nitroimidazole derivatives, including racemates, S-type isomers and R-type isomers. The structural formula is as follows:
Figure RE-GDA0002384363250000023
Figure RE-GDA0002384363250000031
the invention aims to provide nitroimidazole derivatives, which comprise pharmaceutically acceptable medicinal salts, solvate or hydrate. Preferred compounds have the following structural formula:
Figure RE-GDA0002384363250000032
Figure RE-GDA0002384363250000041
the nitroimidazole derivative is realized by the following steps:
adding a solvent into a reaction bottle, adding 2-methyl-5-nitroimidazole, cooling to 0-5 ℃, dropwise adding S- (+) -epoxy chloropropane, stopping the reaction after 35 hours of dropwise reaction, heating to 85-95 ℃, concentrating under reduced pressure, dissolving concentrated residues in water, adjusting the pH value to 3-4, filtering, extracting filtrate with acid water, adjusting the pH value of an acid water layer to 7-8 with ammonia water, stirring, crystallizing, filtering and drying to obtain the levoornidazole. Adding the levoornidazole, the organic solvent and the phosphorus oxychloride into a reaction bottle. Starting stirring, heating to react for 4-8 hours, concentrating under reduced pressure until no liquid drops are generated after the reaction is finished, putting the residue into a chromatographic column, eluting with an organic solvent, concentrating the target eluent to 1/3 of the total volume after the elution is finished, cooling to-5-0 ℃, and crystallizing for 8-12 hours. Filtering, and placing the filter cake in a blast drying oven, controlling the temperature to be 20-30 ℃, and drying for 2-4 hours to obtain the nitroimidazole derivative shown as the formula I.
Figure RE-GDA0002384363250000051
In the preparation method of the nitroimidazole derivative, the organic solvent is acetonitrile, ethyl acetate, dichloromethane, tetrahydrofuran and the like.
In the preparation method of the nitroimidazole derivative, the molar ratio of the levoornidazole to the phosphorus oxychloride is 2: 0.1-2.
The reaction temperature in the preparation method of the nitroimidazole derivatives is 40-90 ℃.
The elution solvent in the preparation method of the nitroimidazole derivative is one or more than two of dichloromethane, methanol, ethyl acetate, toluene, acetone, ethanol, isopropanol and other solvents which are mixed in any proportion.
The crystallization temperature in the preparation method of the nitroimidazole derivatives is-20 ℃ to 5 ℃.
The nitroimidazole derivative can be used independently or prepared into medicine or pharmaceutical composition, and comprises pharmaceutically acceptable carriers and auxiliary materials besides active ingredients.
The pharmaceutical composition of the nitroimidazole derivatives provided by the invention is tablets, suppositories, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated agents, granules, dry powders, oral solutions, small injection needles, freeze-dried injection powder for injection or large infusion solutions.
The auxiliary materials of the nitroimidazole derivatives are one or more selected from the following materials: pH adjusting agents, diluents, solubilizers, disintegrants, suspending agents, lubricants, binders, fillers, flavoring agents, sweeteners, antioxidants, surfactants, preservatives, encapsulating agents, and pigments.
The nitroimidazole derivative disclosed by the invention can be prepared into a high-purity product, and has good solubility and unexpected stability. Has superior stability compared with the disclosed medicinal salt.
The excellent stability of the nitroimidazole derivative enables the nitroimidazole derivative to be placed for a long time when being used for oral administration, and the effective period is prolonged.
The excellent stability of the nitroimidazole derivative enables the nitroimidazole derivative to be stored for a long time without harsh storage conditions, so that the nitroimidazole derivative can be stored for a long time at room temperature. Compared with the disclosed medicinal salt, the compound has the advantage of storage at the temperature below 15 ℃.
Drawings
Fig. 1 shows the amount of levoornidazole measured in the blood.
FIG. 2 shows the measured amounts of the mono-sodium salt of nitroimidazole compound II and the comparative compound in blood.
Detailed Description
The present invention will be described in further detail with reference to examples, but it should be understood that the scope of the present invention is not limited to these examples.
Example 1: preparation of nitroimidazole compounds II
139.4g of formic acid and 13.3g of concentrated sulfuric acid were added to a reaction flask, and stirring was started, followed by addition of 40.0g of 2-methyl-5-nitroimidazole. Controlling the temperature of the reaction liquid to be 0-5 ℃, and dripping 89.8g of S- (+) -epichlorohydrin. After the dropwise addition, the temperature is raised to 5-10 ℃, and the reaction is stopped after stirring and reacting for 35 hours. Heating to 85-95 deg.c, decompression concentrating until no liquid flows out, and stopping concentrating. Adding 190g of purified water into the concentrated residue, cooling to 0-5 ℃, then dropwise adding ammonia water to adjust the pH value to 3-4, and continuing stirring for 2 hours after the pH value is stable. And (4) filtering by shaking, discarding a filter cake, and pumping the filtrate into a 100mL reaction bottle. The filtrate was extracted with dichloromethane (135g × 3), the aqueous layer was discarded, the combined dichloromethane layers were extracted into a 100mL reaction flask, extracted with 10% hydrochloric acid solution (about 60g × 2), and the combined acid aqueous layers were collected. And (4) dropwise adding ammonia water, and adjusting the pH value to 7-8. And then naturally cooling to 25-30 ℃, stirring and crystallizing for 2-4 hours, then cooling to 5-10 ℃, and stirring and crystallizing for 4-8 hours. Then, filtering by throwing, drying a filter cake at 40-50 ℃ to obtain the levoornidazole, adding 50g of the levoornidazole, 100ml of acetonitrile and 17g of phosphorus oxychloride into a reaction bottle, starting stirring, heating to reflux and react for 4-8 hours, after the reaction is finished, concentrating under reduced pressure until no liquid drops exist, putting the residue into a chromatographic column, and adding dichloromethane: and (3) eluting with methanol (2:1), concentrating the target eluent to 1/3 of the total volume after the elution is finished, cooling to-5-0 ℃, and crystallizing for 8-12 hours. Filtering, and placing a filter cake in a blast drying oven, controlling the temperature to be 20-30 ℃, and drying for 2-4 hours to obtain 40.6g of the nitroimidazole compound II with the yield of 81.2 percent.
Figure RE-GDA0002384363250000061
Example 2: preparation of nitroimidazole compounds II
100ml acetonitrile, 17g phosphorus oxychloride and 50g levoornidazole were added to the reaction flask. Starting stirring, heating to reflux reaction for 4-8 hours, after the reaction is finished, concentrating under reduced pressure until no liquid drops exist, putting the residue into a chromatographic column, and reacting with dichloromethane: and (3) eluting with methanol (2:1), concentrating the target eluent to 1/3 of the total volume after the elution is finished, cooling to-5-0 ℃, and crystallizing for 8-12 hours. Filtering, and placing a filter cake in a blast drying oven, controlling the temperature to be 20-30 ℃, and drying for 2-4 hours to obtain 40.3g of the nitroimidazole compound II with the yield of 80.6%.
Example 3: preparation of racemate of nitroimidazole compound II
100ml acetonitrile, 17g phosphorus oxychloride and 50g ornidazole were added to the reaction flask. Starting stirring, heating to reflux reaction for 4-8 hours, after the reaction is finished, concentrating under reduced pressure until no liquid drops exist, putting the residue into a chromatographic column, and reacting with dichloromethane: and (3) eluting with methanol (2:1), concentrating the target eluent to 1/3 of the total volume after the elution is finished, cooling to-5-0 ℃, and crystallizing for 8-12 hours. Filtering, and placing a filter cake in a blast drying oven, controlling the temperature to be between 20 and 30 ℃, and drying for 2 to 4 hours to obtain 43.5g of the racemate of the nitroimidazole compound II with the yield of 87.0 percent.
Example 4: preparation of D-isomer of nitroimidazole compound II
100ml acetonitrile, 17g phosphorus oxychloride and 50g dexornidazole were added to the reaction flask. Starting stirring, heating to reflux reaction for 4-8 hours, after the reaction is finished, concentrating under reduced pressure until no liquid drops exist, putting the residue into a chromatographic column, and reacting with dichloromethane: and (3) eluting with methanol (2:1), concentrating the target eluent to 1/3 of the total volume after the elution is finished, cooling to-5-0 ℃, and crystallizing for 8-12 hours. Filtering, and placing a filter cake in a blast drying oven, controlling the temperature to be between 20 and 30 ℃, drying for 2 to 4 hours, wherein the yield of the nitroimidazole compound II dextroisomer is 77.2 percent and 38.6 g.
Example 5: preparation of nitroimidazole compound II sodium salt
Adding 200ml of acetonitrile, 17g of phosphorus oxychloride and 50g of levonidazole into a reaction bottle, starting stirring, heating to 45-55 ℃ for reaction for 4-8 hours, after the reaction is finished, adding 50g of purified water, heating to 60-70 ℃, hydrolyzing for 2-4 hours, concentrating under reduced pressure to dryness, adding 1000ml of ethanol into residues for dissolving, filtering, adjusting the pH of filtrate to 7-8 with 30% sodium hydroxide, filtering, concentrating the filtrate under reduced pressure to dryness, heating and dissolving the residues to 50-60 ℃ with 200ml of methanol/ethyl acetate (1/4), adding 2g of activated carbon after clearing, stirring and decoloring for 10 minutes, filtering, cooling the filtrate to-5-0 ℃, stirring and crystallizing for 2-4 hours, filtering, drying filter cakes for 2-4 hours at 20-30 ℃, obtaining 32.6g of sodium salt of the nitroimidazole compound II, and obtaining the yield of 65.2%.
Example 6: preparation of choline salt of nitroimidazole compound II
Adding 200ml of acetonitrile, 17g of phosphorus oxychloride and 50g of levonidazole into a reaction bottle, starting stirring, heating to 45-55 ℃ for reaction for 4-8 hours, after the reaction is finished, adding 50g of purified water, heating to 60-70 ℃, hydrolyzing for 2-4 hours, concentrating under reduced pressure to be dry, adding 1000ml of ethanol into residues for dissolving, filtering, adjusting the pH of filtrate to 7-8 with 30% choline hydroxide, filtering, concentrating the filtrate under reduced pressure to be dry, heating and dissolving the residues to 50-60 ℃ with 150ml of ethanol/water (2/1), adding 1.5g of activated carbon after clearing, stirring and decoloring for 10 minutes, filtering, cooling the filtrate to-5-0 ℃, stirring and crystallizing for 2-4 hours, filtering, drying filter cakes for 2-4 hours at 20-30 ℃, obtaining 39.5g of nitroimidazole compound II choline salt, and obtaining the yield of 79.0%.
Example 7: preparation of racemate sodium salt of nitroimidazole compound II
Adding 200ml of acetonitrile, 17g of phosphorus oxychloride and 50g of ornidazole into a reaction bottle, starting stirring, heating to 45-55 ℃ for reaction for 4-8 hours, after the reaction is finished, adding 50g of purified water, heating to 60-70 ℃, hydrolyzing for 2-4 hours, concentrating under reduced pressure to be dry, adding 1000ml of ethanol into residues for dissolving, filtering, adjusting the pH of filtrate to 7-8 with ammonia water, filtering, concentrating the filtrate under reduced pressure to be dry, heating to 50-60 ℃ with 200ml of ethyl acetate/methanol (3/1) to dissolve the residues, adding 2g of activated carbon after clearing, stirring and decoloring for 10 minutes, filtering, cooling the filtrate to-5-0 ℃, stirring and crystallizing for 2-4 hours, filtering, drying a filter cake at 20-30 ℃ for 2-4 hours to obtain 37.7g of racemate sodium salt of the nitroimidazole compound II, and the yield is 75.4%.
Example 8: preparation of hexahydrate of sodium salt of nitroimidazole compound II
Adding 200nl of 75% ethanol into a reaction bottle, adding 20g of nitroimidazole compound II sodium salt, heating to 60-70 ℃, stirring for dissolving, adding 2g of activated carbon, stirring for 20 minutes, filtering, slowly cooling the filtrate to-5-0 ℃, crystallizing, filtering, and drying the filter cake at 40-50 ℃ to obtain 15.3g of hexahydrate of the nitroimidazole compound II sodium salt, wherein the yield is 76.5%.
Example 9: preparation of heptahydrate of sodium salt of nitroimidazole compound II
Adding 40ml of purified water into a reaction bottle, adding 20g of nitroimidazole compound II sodium salt, stirring at room temperature to dissolve the mixture clearly, adding 2g of activated carbon, stirring for 20 minutes, filtering, slowly controlling the temperature of the filtrate to be 5-10 ℃, slowly dropwise adding 300ml of absolute ethyl alcohol, stirring for 2-4 hours after dropwise adding, filtering, and drying the filter cake at 40-50 ℃, namely 18.2g of heptahydrate of the nitroimidazole compound II sodium salt, wherein the yield is 91.0%.
Example 10: preparation of nitroimidazole compound II sodium salt tablet
Prescription:
nitro imidazole compound II (sodium salt) 12.5g
Hydroxypropyl methylcellulose 5.97g
Starch 6.55g
Lactose 0.75g
Sodium carbonate 0.12g
Magnesium stearate 0.25g
5% Povidone 0.85g
The preparation method comprises the following steps: taking the nitroimidazole compound II sodium salt, crushing and sieving by a 80-mesh sieve; weighing and uniformly mixing the nitroimidazole compound II sodium salt with the hydroxypropyl methyl cellulose, the starch, the lactose and the sodium carbonate according to the prescription amount. Preparing the materials into soft materials by using 5% povidone K30 solution, granulating by using a 20-mesh sieve, and drying at 40-60 ℃ until the water content in the granules is about 5%. Sieving with a 20-mesh sieve, grading, adding magnesium stearate according to the formula amount, mixing, measuring the content of an intermediate, and determining the weight of tablets; and (6) tabletting.
Example 11: preparation of nitroimidazole compound II sodium salt granules
Prescription:
nitro imidazole compound II sodium salt 12.5g
Starch 6.55g
Sucralose 10.0g
5% Povidone 1.0g
The preparation method comprises the following steps: taking the nitroimidazole compound II sodium salt, crushing and sieving by a 80-mesh sieve; weighing the starch and the sucralose according to the prescription amount, and mixing uniformly. Preparing the materials into soft materials by using 5% povidone K30 solution, granulating by using a 20-mesh sieve, and drying at 40-60 ℃ until the water content in the granules is about 5%. Sieving with 20 mesh sieve, grading, measuring intermediate content, and packaging.
Example 12: preparation of nitroimidazole compound II sodium salt for injection
Prescription:
nitro imidazole compound II sodium salt 50g
pH regulator 4.0-9.0
Mannitol 20g
Water (W) 200ml
The preparation method comprises the following steps: adding a batch volume of water for injection, weighing the nitroimidazole compound II sodium salt and mannitol according to the prescription amount, stirring to fully dissolve, adjusting the pH to 4.0-9.0 by using a pH regulator, filtering by using a 0.22um microporous membrane, and filling; freeze drying, capping and packaging.
EXAMPLE 13 preparation of nitroimidazole Compound II for injection
Prescription:
nitroimidazole compounds II 50g
pH regulator 4.0-9.0
Water for injection 500ml
The preparation method comprises the following steps: adding 450ml of water for injection, weighing the nitroimidazole compound II according to the prescription amount, stirring to fully dissolve, adjusting the pH to 4.0-9.0 by using a pH regulator, supplementing purified water to the full amount, adding 0.05% of active medicinal carbon, stirring for 30 minutes, filtering by using a 0.22um microporous membrane, and filling; and (6) sealing, sterilizing and packaging.
Experimental example 1: acute toxicity test of Nitroimidazole Compounds of the invention and control Compounds on intravenous administration to mice
To test the acute toxicity of the compounds of the invention and the comparative compounds, the following experiments were performed.
The compound of the present invention was dissolved in water and administered to 5 ICR mice (5-week-old, male, mice weighing 20 g. + -.2 g). Intravenous administration to determine median Lethal Dose (LD)50Mg/kg). Levoornidazole was used as control. The results are shown in the following table.
Compound (I) Median Lethal Dose (LD)50，mg/kg)
Levoornidazole 332
Nitroimidazole compounds II 487
Nitro imidazole compound II sodium salt 493
Nitroimidazole compound II choline salt 503
Nitro imidazole compound II sodium salt heptahydrate 542
The test result shows that: the LD50 value of the compound is higher than that of the levoornidazole, which indicates that the nitroimidazole compound has better safety than the levoornidazole.
Experimental example 2: experiment of intravenous administration of the compound of the invention on resisting bacteroides fragilis vaginitis
1. Experimental Material
1.1 Experimental instruments
A blood cell counting plate, a paraffin slicer, an SPX-250B biochemical incubator, an ultra-purification workbench, a microsyringe, a pressure steam sterilizer, an optical microscope and an electronic analytical balance.
1.2 Experimental reagents
Estradiol benzoate injection, polyethylene glycol and Bacteroides fragilis activation culture broth.
1.3 Experimental animals
KM mice, 18-22g in weight, female, were provided by the laboratory animals center of Jiangsu province.
1.4 test strains
Standard Bacteroides fragilis was purchased from American type culture Collection under the strain number ATCC 25285.
2. Experimental methods
The mice are randomly grouped after being weighed: a levoornidazole group, a test compound group and a solvent group, wherein each group comprises 20. Before infection with Bacteroides fragilis, animals in each group were given 0.5ml of estradiol benzoate (2mg/ml) subcutaneously for 6 consecutive days to enter estrus, and thereafter injected 1 time every 2 days until the experiment was completed. After 6 days, 20ul of a 3.5 x 10 concentration was vaginal injected into each mouse6CFU/m1, a fragile bacteroidal liquid, resulting in a vaginal infection model. From the first day after infection, the tail vein of each group of animals is administered with the corresponding drug 20mg/kg (calculated as levoornidazole), and the drug volume is administered with 0.1ml/kg once a day for 5 consecutive days, and the model group is administered with the same volume of solvent (physiological saline). On the 3 rd and 5 th days after infection, the vagina of the mouse is wiped by a sterile cotton swab, the cotton swab is soaked in 0.9ml of main saline, the bacterial liquid is diluted into a series of concentrations by 10 times of increment, then 100ul of bacterial liquid with each concentration is respectively taken and inoculated on the bacteroides fragilis activation culture broth, and the fungal load of the bacteroides fragilis on the vagina is observed.
3. Results of the experiment
Bacteroides fragilis vaginitis (intravenous administration): vaginal fungus load of each group of mice
Figure RE-GDA0002384363250000111
Note: data are presented as mean ± standard deviation of the logarithm of CFU values for 20 mice.
The experimental results show that the fungus load of mice in the compound group is obviously reduced compared with that of a solvent group after 5 days of intravenous administration, and the compound is superior to that of a levoornidazole group, so that an obvious curative effect is achieved.
Example 3: experiment of compound of the invention for resisting bacteroides fragilis vaginitis by intragastric administration
1. Experimental Material
1.1 Experimental instruments
A blood cell counting plate, a paraffin slicer, an SPX-250B biochemical incubator, an ultra-purification workbench, a microsyringe, a pressure steam sterilizer, an optical microscope and an electronic analytical balance.
1.2 Experimental reagents
Estradiol benzoate injection, polyethylene glycol and Bacteroides fragilis activation culture broth.
1.3 Experimental animals
KM mice, 18-22g in weight, female, were provided by the laboratory animals center of Jiangsu province.
1.4 test strains
The standard strain, Bacteroides fragilis, was purchased from American type culture Collection and has the strain number ATCC 25285.
2. Experimental methods
The mice are randomly grouped after being weighed: a levoornidazole group, a test compound group and a solvent group, wherein each group comprises 20. Before infection with Bacteroides fragilis, animals in each group were given 0.5ml of estradiol benzoate (2mg/ml) subcutaneously for 6 consecutive days to enter estrus, and thereafter injected 1 time every 2 days until the experiment was completed. After 6 days, 20ul of 3.5 xl 0 was vaginally injected into each mouse6CFU/m1, a fragile bacteroidal liquid, resulting in a vaginal infection model. From the first day after infection, each group of animals is administered with corresponding drug 20mg/kg (calculated by levoornidazole) via intragastric administration, the administration volume is 0.1ml/kg, once a day for 15 days continuously, and the administration is performed by model group, etcVolume solvent (normal saline). On the 3 rd, 5 th, 7 th, 11 th and 15 th days after infection, the vagina of each group of mice was swabbed with a sterile cotton swab, the cotton swab was soaked in 0.9ml of main saline, the bacterial solution was diluted to a series of concentrations by 10-fold increase, then the bacterial solution of each concentration of l00ul was inoculated on the bacteroides fragilis activation medium broth, and the amount of the fungal load of bacteroides fragilis on the vagina was observed.
3. Results of the experiment
Bacteroides fragilis vaginitis (gavage): vaginal fungus load of each group of mice
Figure RE-GDA0002384363250000112
Figure RE-GDA0002384363250000121
The experimental results show that after the compound is taken for 15 days, the fungus load capacity of mice in the compound group dissolved by normal saline is obviously reduced compared with that of a solvent group, and is superior to that of a levoornidazole group, so that an obvious curative effect is achieved.
Example 4: in vivo pharmacokinetic testing
The test method comprises the following steps: the experimental animal is male beagle dog with weight of 9.9-11.2kg, and is purchased from Beijing Weilitong Hua experimental animal technology GmbH. The groups were randomized into 3 groups of 3 animals based on beagle body weight. The dose of administration for each group was 30mg/kg (calculated as levoornidazole) and the route are given in the table below.
Group of Medicine Dosage (mg/kg) Pathway(s) Number of
Group 1 Levoornidazole 30 Vein 3
Group 2 Mono-sodium nitroimidazole compound II 30 Vein 3
Group 3 Sodium levoornidazole phosphate monosodium salt 30 Vein 3
Beagle dogs were fasted for 12 hours prior to pharmacokinetic testing. A single dose of the compound was then administered intravenously as shown in the table above. Collecting 2ml of blood at the administration initiation timing by means of intravenous injection in the foreleg, wherein for the intravenously administered animal groups, blood was collected at 0, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, and 24 hours after administration; urine was collected at 2 hours, 4 hours, 8 hours, 12 hours, 24 hours. The blood samples were collected in sample tubes with EDTA, immediately centrifuged at 4000rpm for 5 minutes at 4 ℃, and then the plasma was transferred to another sample tube and stored at-20 ℃.
The samples were subjected to pharmacokinetic testing by measuring the concentration of levoornidazole formed by the conversion of the test compound in the blood and urine samples taken at each time point, using the following methods and instruments:
HPLC：Shimadzu
MS：AB API4000Q
column: phenomenex Luna 5um C18
Mobile phase: 100% acetonitrile (3mM ammonium acetate) and 100% water (3mM ammonium acetate)
The quantitative method comprises the following steps: internal standard method
The results of the in vivo pharmacokinetic experiments were as follows:
measurement unit of levoornidazole in superficial blood: ng (ng)
Figure RE-GDA0002384363250000131
In the table blood, the measurement unit of the mono-sodium salt of the nitroimidazole compound II and the comparative compound is as follows: ng (ng)
Figure RE-GDA0002384363250000132
The experimental results show that the compound of the invention achieves steady-state blood concentration better than that of the levoornidazole and the comparative compound, the compound of the invention has slower metabolic speed, and can achieve higher stable blood concentration due to the slow release process, and the data show that the compound of the invention is better than that of the levoornidazole and the comparative compound.
Experimental example 5: stability comparison research of heptahydrate of nitroimidazole compound II sodium salt and levoornidazole phosphate disodium hydrate
The results of comparing the stability of the heptahydrate of the sodium salt of the nitroimidazole compound ii prepared in the above example 9 with that of the disodium levoornidazole phosphate hydrate at 60 ℃ for 10 days are as follows:
chromatographic conditions are as follows:
mobile phase A: potassium dihydrogen phosphate 6.8g/L, sodium heptanesulfonate 0.3g/L, and triethylamine is used for adjusting the pH value to 6.5.
Mobile phase B: methanol
Detection wavelength: 321 nm; flow rate: l.0ml/min; column temperature: 25 ℃; sample concentration: 2mg/ml
Dilution medium: 0.1mol/L potassium dihydrogen phosphate
Gradient conditions:
Figure RE-GDA0002384363250000133
Figure RE-GDA0002384363250000141
results comparing stability of the present Compounds with that of the comparative Compounds
Figure RE-GDA0002384363250000142
The above experimental results show that the compound of the present invention and the control compound are left at 60 ℃ for 10 days, the compound of the present invention has almost no degradation of impurities, and the control compound has significant degradation of impurities after 10 days, so the compound of the present invention is superior to the control compound.
The above examples are only illustrative of several embodiments of the present invention, but should not be construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. The protection scope of the present invention should be subject to the appended claims.

Claims (6)

1. Nitroimidazole derivative of formula I
Figure 282600DEST_PATH_IMAGE001
I
Wherein: n is 0 to 12, preferably 0 to 8, more preferably 0 to 6;
r represents hydrogen, alkali metal, amino acid, choline and meglumine;
x is water, methanol, ethanol, acetone, ethyl acetate, isopropanol, etc.
2. A nitroimidazole derivative according to claim 1, wherein the compound comprises S, racemate, R isomers.
3. Nitroimidazole derivatives according to claim 1, characterized in that the preparation process of said compounds comprises the following steps:
(a) reacting a compound shown in a formula A with a compound shown in a formula B in a solvent to form a compound shown in a formula C;
Figure 249288DEST_PATH_IMAGE002
(b) reacting the compound shown in the formula C formed in the step (a) with the compound shown in the formula D in an organic solvent in the presence of inert gas to obtain the compound shown in the formula I;
Figure 270596DEST_PATH_IMAGE003
。
4. the method for producing a nitroimidazole derivative according to claim 3, wherein the molar ratio of the compound C to the compound D is 2:0.1 to 2.
5. A pharmaceutical composition comprising a compound of claim 1 as an active ingredient and a pharmaceutically acceptable carrier.
6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is used for treating diseases caused by anaerobic bacterial infection or protozoal infection, and for preventing anaerobic bacterial infection or protozoal infection.
CN201911075152.7A 2019-11-06 2019-11-06 Nitroimidazole derivative and preparation method and application thereof Active CN112778364B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201911075152.7A CN112778364B (en) 2019-11-06 2019-11-06 Nitroimidazole derivative and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201911075152.7A CN112778364B (en) 2019-11-06 2019-11-06 Nitroimidazole derivative and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN112778364A true CN112778364A (en) 2021-05-11
CN112778364B CN112778364B (en) 2023-12-08

Family

ID=75747574

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911075152.7A Active CN112778364B (en) 2019-11-06 2019-11-06 Nitroimidazole derivative and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN112778364B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478494A (en) * 2020-10-26 2022-05-13 南京锐志生物医药有限公司 Derivative of nitroimidazole compound, pharmaceutical composition, preparation method and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102516299A (en) * 2011-12-09 2012-06-27 陕西合成药业有限公司 S-Ornidazole phosphate amino acid salt, its preparation method and application
CN105646580A (en) * 2016-03-04 2016-06-08 中山福运生物科技有限公司 Method for producing pentahydrate s-ornidazole disodium phosphate
CN107235911A (en) * 2016-03-28 2017-10-10 陕西合成药业股份有限公司 A kind of purposes of 2- methyl-4-nitro iminazoles derivative and preparation method thereof and Pharmaceutical composition
CN107365272A (en) * 2016-05-12 2017-11-21 陕西合成药业股份有限公司 A kind of novel imidazole class compound and preparation method thereof and application medically

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102516299A (en) * 2011-12-09 2012-06-27 陕西合成药业有限公司 S-Ornidazole phosphate amino acid salt, its preparation method and application
CN105646580A (en) * 2016-03-04 2016-06-08 中山福运生物科技有限公司 Method for producing pentahydrate s-ornidazole disodium phosphate
CN107235911A (en) * 2016-03-28 2017-10-10 陕西合成药业股份有限公司 A kind of purposes of 2- methyl-4-nitro iminazoles derivative and preparation method thereof and Pharmaceutical composition
CN107365272A (en) * 2016-05-12 2017-11-21 陕西合成药业股份有限公司 A kind of novel imidazole class compound and preparation method thereof and application medically

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478494A (en) * 2020-10-26 2022-05-13 南京锐志生物医药有限公司 Derivative of nitroimidazole compound, pharmaceutical composition, preparation method and application thereof
CN114478494B (en) * 2020-10-26 2023-10-27 南京锐志生物医药有限公司 Derivatives of nitroimidazole compounds, pharmaceutical composition, preparation method and application thereof

Also Published As

Publication number Publication date
CN112778364B (en) 2023-12-08

Similar Documents

Publication Publication Date Title
EP2749276B1 (en) Use of patchouli alcohol in preparation of drug against helicobacter pylori
JP2009502752A (en) Water-soluble artemisinin derivative, preparation method thereof, pharmaceutical composition and use
CN102040592B (en) Coumarin azole compound with antimicrobial activity, and preparation method and medicinal application thereof
US8217013B2 (en) Polyene diester antibiotics
CN100494193C (en) 8-octyl berberine hydrochloride and its synthesis process and application
CN112778364B (en) Nitroimidazole derivative and preparation method and application thereof
CN102918051B (en) N6-(methyl ferrocene) quinazoline-2,4,6-triamine (H2) and its derivative, and the prodrug being used as antiseptic-germicide, antiparasitic, protozoacide and anti-Leishmania agent
CN112778363B (en) Nitroimidazole derivative and preparation method and application thereof
CN112500344B (en) Crystalline form of roxasistat and preparation method thereof
CN107365272A (en) A kind of novel imidazole class compound and preparation method thereof and application medically
CN112778369B (en) Triazole derivative and preparation method and application thereof
DE3031788C2 (en)
CN101381386B (en) Lactose-azithromycin hydrate, preparation and use thereof
CN101386593B (en) Norfloxacin glutamate hydrate and preparation and use thereof
CN111514151B (en) Application of three compounds in myrobalan in preparing anti-inflammatory drugs and preparation method thereof
JP7026264B2 (en) Posaconazole phosphate monocholine salt, its preparation method and use
JPS6412277B2 (en)
CN1281447A (en) (i(E))-3-[1-i(n)-butyl-5-[2-(-carboxyphenyl) methoxy-4-chlorophenyl]-1i(H)-pyrazol-4-yl]-2-[(5-methoxy-2,3-dihydrobenzofuran-6-yl) methyl]-prop-2-enoic acid monoargininyl salt
CN107556304B (en) Nitroimidazole medicine and preparation method and application thereof
CN103159710B (en) Antiviral decalin derivate
JPH04139180A (en) New xanthones and aldose reductase inhibitor comprising the same xanthones as active ingredient
CN114907405A (en) Deuterated nitroimidazole compound and pharmaceutical application thereof
RU2181051C1 (en) Method to obtain injection preparation based upon a substance of p-vitamin activity
JPH0759515B2 (en) Aldo-reductase inhibitor
CN101161652A (en) Quinolizine derivatives having antibacterial activity

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant