CN103830164A - Moxifloxacin hydrochloride injection liquid and preparation method thereof - Google Patents
Moxifloxacin hydrochloride injection liquid and preparation method thereof Download PDFInfo
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- CN103830164A CN103830164A CN201210472535.XA CN201210472535A CN103830164A CN 103830164 A CN103830164 A CN 103830164A CN 201210472535 A CN201210472535 A CN 201210472535A CN 103830164 A CN103830164 A CN 103830164A
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Abstract
The present invention discloses a moxifloxacin hydrochloride injection liquid and a preparation method thereof, wherein the moxifloxacin hydrochloride injection liquid comprises a solubilizer and a metal ion complexing agent.
Description
Field
The disclosure relates to field of pharmaceutical preparations, and more specifically, the disclosure relates to moxifloxacin hydrochloride injection and preparation method thereof.
Background
Moxifloxacin hydrochloride (Moxifloxacin) is the super wide spectrum quinolones of Bayer A.G's development, the commodity multiple pleasure (Avelox) of visiing by name.Moxifloxacin hydrochloride structural formula is as follows.
Moxifloxacin hydrochloride demonstrates in vitro has broad spectrum antibiotic activity to gram positive bacteria, gram-negative bacteria, anaerobe, acid fast bacteria and the atypical microorganism such as mycoplasma, chlamydia and legionella, and its antibacterial mechanisms is for disturbing II, IV topoisomerase.Moxifloxacin hydrochloride is active high in vivo, and absolute bioavailability amounts to approximately 91%, and its medication is not affected by feed.
At present, the less stable of commercially available moxifloxacin hydrochloride injection in transport and storing process, for example, has precipitation when transporting or storing and occur under 2 ° of C-8 ° of C.And in preparation process, often there is the situation of color burn or variable color in moxifloxacin hydrochloride solution.In addition, because commercially available moxifloxacin hydrochloride injection dosage is large, so the poor operability in the process of producing, production, transport and carrying cost are high, and amount of infusion brings inconvenience greatly also to clinical application and patient simultaneously.
General introduction
The disclosure relates in one aspect to moxifloxacin hydrochloride injection, based on the volume of described injection, and the complexing of metal ion agent of the moxifloxacin hydrochloride that comprises 1-2W/V, the solubilizing agent of 0.005-0.6W/V and 0.001-0.1W/V.
Another aspect of the present disclosure relates to the method for preparing moxifloxacin hydrochloride injection, and it comprises: water for injection is provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Optionally, then add pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0.
Describe in detail
Unless separately indicated in the disclosure, in whole description and claims, word " comprises " and " comprising " should be interpreted as meaning open, that include formula, " includes but not limited to ".
The term adopting " W/V " in the disclosure is weight/volume, and it represents the quantity that unit of weight in 100ml volume is gram.For example 1W/V represents to contain the moxifloxacin hydrochloride that weight is 1g in the moxifloxacin hydrochloride injection of 100ml.
The disclosure relates in one aspect to moxifloxacin hydrochloride injection, based on the volume of described injection, and the complexing of metal ion agent of the moxifloxacin hydrochloride that comprises 1-2W/V, the solubilizing agent of 0.005-0.6W/V and 0.001-0.1W/V.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.005-0.1W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.01-0.6W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.01-0.1W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.01-0.025W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.025-0.1W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.005-0.01W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.01-0.025W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.025-0.1W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.1-0.6W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, moxifloxacin hydrochloride infusion pump is 0.005W/V, 0.01W/V, 0.025W/V, 0.1W/V or 0.6W/V containing the amount of solubilizing agent.
In other embodiments, based on the volume of described injection, the amount of moxifloxacin hydrochloride infusion pump metal ion chelating agent is 0.001-0.01W/V.
In other embodiments, based on the volume of described injection, the amount of moxifloxacin hydrochloride infusion pump metal ion chelating agent is 0.01-0.1W/V.
In other embodiments, based on the volume of described injection, the amount of moxifloxacin hydrochloride infusion pump metal ion chelating agent is 0.001W/V, 0.01W/V or 0.1W/V.
In some embodiments, conventional solubilizing agent in injection preparation is selected in solubilizing agent.
The example that can be used in exemplary solubilizing agent of the present disclosure includes but not limited to polyoxyethylene castor oil, polyoxyethylene ether (60) castor oil hydrogenated, Liquid Macrogol, PEG400, HS15, Polyethylene Glycol-15-hydroxy stearic acid ester, PLURONICS F87 and Tween 80.
In some embodiments, wherein solubilizing agent is PLURONICS F87.
In some embodiments, wherein solubilizing agent is Polyethylene Glycol-15-hydroxy stearic acid ester.
In some embodiments, wherein solubilizing agent is Tween 80.
The example that can be used in exemplary complexing of metal ion agent of the present disclosure includes but not limited to cyclohexanediamine four sodium acetates, N-hydroxy-ethylenediamine three acetic acid, sodium calcium edetate, disodium edetate and edetic acid.
In some embodiments, wherein complexing of metal ion agent is sodium calcium edetate.
In some embodiments, wherein complexing of metal ion agent is disodium edetate.
In some embodiments, wherein complexing of metal ion agent is edetic acid.
The scope of the pH value of moxifloxacin hydrochloride is between 4.0-4.6, and its stability within the scope of this pH value is also best conventionally.But in injection preparation process, the many factors that may exist, such as water for injection, can cause the pH value of injection solvent to depart from the best stabilized interval of moxifloxacin hydrochloride.Therefore in the time preparing moxifloxacin hydrochloride injection, the pH value of injection need to be adjusted within the scope of this.
Conventionally, regulate pH value by adding acidic ph modifier to solution.If the excessive acid adding, also can add alkaline pH regulator again.
In the disclosure, for improving the stability of product, add pH adjusting agent so that pH value is adjusted between 4.0-5.0 to moxifloxacin hydrochloride solution.
The example that can be used in exemplary pH adjusting agent of the present disclosure includes but not limited to hydrochloric acid, sulphuric acid, phosphoric acid, acetic acid, malic acid, lactic acid, citric acid and tartaric acid.
In some embodiments, pH adjusting agent is hydrochloric acid.
The example that can be used in exemplary pH adjusting agent of the present disclosure includes but not limited to sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, sodium tartrate and sodium lactate.
In some embodiments, pH adjusting agent is sodium hydroxide.
The disclosure relates to the method for preparing moxifloxacin hydrochloride injection on the other hand, and the method comprises: water for injection is provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Additive is dissolved completely; Add again pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0.
In some embodiments, the method for preparing moxifloxacin hydrochloride injection is disclosed.The method comprises the following steps: the water for injection that the 75%-95% of cumulative volume is provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Being incubated 60 ° of C-80 ° of C and stirring dissolves additive completely; Add water for injection to cumulative volume.
In some embodiments, the method for preparing moxifloxacin hydrochloride injection is disclosed.The method comprises the following steps: the water for injection that the 75%-95% of cumulative volume is provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Being incubated 60 ° of C-80 ° of C and stirring dissolves additive completely; Add again pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0; Add water for injection to cumulative volume; Add injection remove impurity with active carbon, filter and remove active carbon; Fine straining, fill, hot pressing steam sterilizing.
In some embodiments, the method for preparing moxifloxacin hydrochloride injection is disclosed.The method comprises the following steps: the water for injection that whole volumes are provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Being incubated 60 ° of C-80 ° of C and stirring dissolves additive completely; Add again pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0.
In some embodiments, the method for preparing moxifloxacin hydrochloride injection is disclosed.The method comprises the following steps: the water for injection that whole volumes are provided; Add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection; Add again appropriate complexing of metal ion agent; Being incubated 60 ° of C-80 ° of C and stirring dissolves additive completely; Add again pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0; Add injection active carbon, stir, boil, filter and remove active carbon; Fine straining, fill, hot pressing steam sterilizing.
In some embodiments, moxifloxacin hydrochloride injection of the present disclosure, can be before use according to the clinical sterile solution being injected in vivo that needs dilution (dilute solution is commercially available 0.9% sodium chloride injection and 5% glucose injection) to become variable concentrations.
It will be appreciated by those skilled in the art that the technical scheme described in the respective embodiments described above can further combine and form new technical scheme according to object of the present disclosure.
Hereinafter, the disclosure will be explained in detail to understand better various aspects of the present disclosure and advantage thereof by following embodiment.But, should be appreciated that following embodiment is nonrestrictive and only for some embodiment of the present disclosure is described.
Embodiment
The reagent adopting in following examples of the present disclosure is commercially available prod.
Embodiment 1: the preparation (0.4g ︰ 20ml) of moxifloxacin hydrochloride injection
Formula:
Component | Weight | Content (taking the volume of injection as basis, g/100mL) |
Moxifloxacin hydrochloride | 400g(is in Moxifloxacin) | 2% |
PLURONICS F87 | 120g | 0.6% |
Calcium disodium edetate | 0.2g | 0.001% |
Water for injection adds to 20L, makes 1000 bottles.
Preparation technology:
(1) get qualified moxifloxacin hydrochloride raw material, PLURONICS F87, calcium disodium edetate by above-mentioned formula;
(2) in metal material-compound tank, add the water for injection of batch volumes 80%, add moxifloxacin hydrochloride, PLURONICS F87, the calcium disodium edetate of formula ratio, after being incubated 60 ° of C-80 ° of C and stirring and make fully to dissolve, regulate pH value 4.0-5.0;
(3) add medicinal carbon to stir by 0.05% of cumulative volume, boil 15min, filter decarburization, benefit adds to the full amount of water for injection;
(4) medicinal liquid is crossed respectively the filter membrane fine straining of 0.45 μ m and 0.22 μ m; Intermediate detects qualified rear fill;
(5) the good semi-finished product of fill are sent into sterilizing cabinet sterilizing, sterilising conditions: 121 ° of C hot pressing steams sterilizing 15 minutes;
(6) leak detection, lamp inspection;
(7) finished product is examined qualified rear packaging, warehouse-in entirely.
In this preparation process, there is not the phenomenon of color burn or variable color in moxifloxacin hydrochloride solution.
Embodiment 2: the preparation (0.4g ︰ 20ml) of moxifloxacin hydrochloride injection
Formula:
Water for injection adds to 20L, makes 1000 bottles.
Preparation technology:
(1) get qualified moxifloxacin hydrochloride raw material, Polyethylene Glycol-15-hydroxy stearic acid ester, calcium disodium edetate by above-mentioned formula;
(2) in metal material-compound tank, add the water for injection of batch volumes 95%, add moxifloxacin hydrochloride, Polyethylene Glycol-15-hydroxy stearic acid ester, the disodium edetate of formula ratio, after being incubated 60 ° of C-80 ° of C and stirring and make fully to dissolve, regulate pH value 4.0-5.0;
(3) add medicinal carbon to stir by 0.05% of cumulative volume, boil 15min, filter decarburization, benefit adds to the full amount of water for injection;
(4) medicinal liquid is crossed respectively the filter membrane fine straining of 0.45 μ m and 0.22 μ m; Intermediate detects qualified rear fill;
(5) the good semi-finished product of fill are sent into sterilizing cabinet sterilizing, sterilising conditions: 121 ° of C hot pressing steams sterilizing 15 minutes;
(6) leak detection, lamp inspection;
(7) finished product is examined qualified rear packaging, warehouse-in entirely.
In this preparation process, there is not the phenomenon of color burn or variable color in moxifloxacin hydrochloride solution.
Embodiment 3: the preparation (0.2g ︰ 20ml) of moxifloxacin hydrochloride injection
Formula:
Water for injection adds to 20L, makes 1000 bottles.
Preparation technology:
(1) get qualified moxifloxacin hydrochloride raw material, PLURONICS F87, calcium disodium edetate by above-mentioned formula;
(2) in metal material-compound tank, add the water for injection of batch volumes 80%, add moxifloxacin hydrochloride, PLURONICS F87, the calcium disodium edetate of formula ratio, after being incubated 60-80 ° of C and stirring and make fully to dissolve, regulate pH value 4.0-5.0;
(3) add medicinal charcoal to stir by 0.05% of cumulative volume, boil 15min, filter decarburization, benefit adds to the full amount of water for injection;
(4) medicinal liquid is crossed respectively the filter membrane fine straining of 0.45 μ m and 0.22 μ m; Intermediate detects qualified rear fill;
(5) the good semi-finished product of fill are sent into sterilizing cabinet sterilizing, sterilising conditions: 121 ° of C hot pressing steams sterilizing 15 minutes;
(6) leak detection, lamp inspection;
(7) finished product is examined qualified rear packaging, warehouse-in entirely.
In this preparation process, there is not the phenomenon of color burn or variable color in moxifloxacin hydrochloride solution.
Embodiment 4: the preparation (0.2g ︰ 20ml) of moxifloxacin hydrochloride injection
Formula:
Water for injection adds to 20L, makes 1000 bottles.
Preparation technology:
(1) get qualified moxifloxacin hydrochloride raw material, Polyethylene Glycol-15-hydroxy stearic acid ester, calcium disodium edetate by above-mentioned formula;
(2) in metal material-compound tank, add the water for injection of batch volumes 95%, add moxifloxacin hydrochloride, Polyethylene Glycol-15-hydroxy stearic acid ester, the calcium disodium edetate of formula ratio, after being incubated 60-80 ° of C and stirring and make fully to dissolve, regulate pH value 4.0-5.0;
(3) add medicinal charcoal to stir by 0.05% of cumulative volume, boil 15min, filter decarburization, benefit adds to the full amount of water for injection;
(4) medicinal liquid is crossed respectively the filter membrane fine straining of 0.45 μ m and 0.22 μ m; Intermediate detects qualified rear fill;
(5) the good semi-finished product of fill are sent into sterilizing cabinet sterilizing, sterilising conditions: 121 ° of C hot pressing steams sterilizing 15 minutes;
(6) leak detection, lamp inspection;
(7) finished product is examined qualified rear packaging, warehouse-in entirely.
In this preparation process, there is not the phenomenon of color burn or variable color in moxifloxacin hydrochloride solution.
Embodiment 5: the preparation (0.4g ︰ 20ml) of moxifloxacin hydrochloride injection
Formula:
Water for injection adds to 20L, makes 1000 bottles.
Preparation technology:
(1) get qualified moxifloxacin hydrochloride raw material, Tween 80, calcium disodium edetate by above-mentioned formula;
(2) in metal material-compound tank, add the water for injection of batch volumes 90%, add moxifloxacin hydrochloride, Tween 80, the calcium disodium edetate of formula ratio, after being incubated 60-80 ° of C and stirring and make fully to dissolve, regulate pH value 4.0-5.0;
(3) add medicinal charcoal to stir by 0.05% of cumulative volume, boil 15min, filter decarburization, benefit adds to the full amount of water for injection;
(4) medicinal liquid is crossed respectively the filter membrane fine straining of 0.45 μ m and 0.22 μ m; Intermediate detects qualified rear fill;
(5) the good semi-finished product of fill are sent into sterilizing cabinet sterilizing, sterilising conditions: 121 ° of C hot pressing steams sterilizing 15 minutes;
(6) leak detection, lamp inspection;
(7) finished product is examined qualified rear packaging, warehouse-in entirely.
In this preparation process, there is not the phenomenon of color burn or variable color in moxifloxacin hydrochloride solution.
By above-described embodiment, 1-5 is known, and the specification (Moxifloxacin is heavily measured ︰ injection volume) of moxifloxacin hydrochloride injection of the present disclosure is 0.2g ︰ 20mL to 0.4g ︰ 20mL.
The injection of the aqueous solution form that commercially available moxifloxacin hydrochloride injection is sodium chloride at present.This injection that contains sodium chloride is with large capacity transfusion agent form administration, and its specification is Moxifloxacin 0.4g ︰ 250mL injection.
As can be seen here, the dissolubility of moxifloxacin hydrochloride of the present disclosure in injection solution improved 6-12 doubly, therefore,, in the situation that using equal dosage, disclosure moxifloxacin hydrochloride is significantly less than commercially available moxifloxacin hydrochloride injection at the consumption of injection solution.
Embodiment 6: moxifloxacin hydrochloride injection accelerated stability test research
According to the listed investigation project of stability test of injection in Chinese Pharmacopoeia two annex of version in 2010 " medicine stability test guideline ", moxifloxacin hydrochloride injection is carried out to study on the stability.
Get respectively embodiment 1-5 sample and place 6 months under the condition of 40 ° of C, detected with sampling in 6th month respectively at 1st month, the 2nd month, the 3rd month, the detection data of testing result and 0 day compare, and the results are shown in Table 1.
Table 1 moxifloxacin hydrochloride injection accelerated stability test result
Result shows: moxifloxacin hydrochloride injection, under simulation listing terms of packing, is placed 6 months under the condition of 40 ° of C, every detection index and having no significant change for 0 day.Show that this product has good stability.
Embodiment 7: the application's moxifloxacin hydrochloride injection and the comparison of commercially available moxifloxacin hydrochloride injection particulate matter
Get respectively moxifloxacin hydrochloride injection and the commercially available moxifloxacin hydrochloride injection prepared by the application's method, investigate particulate determination in its freezing-thawing test (light blockage method), both relatively, the results are shown in Table 2.
Table 2 particulate matter is investigation table relatively
As seen from the above table, moxifloxacin hydrochloride injection of the present disclosure is through freezing-thawing test, particulate matter measurement result is better than commercially available moxifloxacin hydrochloride injection, illustrate that the stability under moxifloxacin hydrochloride injection stability, especially low temperature of the present disclosure is better than commercially available moxifloxacin hydrochloride injection.
Although be appreciated that from the foregoing for the object of exemplary illustration and described specific embodiments of the present disclosure, not depart under the condition of spirit and scope of the present disclosure, technical staff can make various distortion or improvement described in this area.These distortion or amendment all should fall into the scope of disclosure claims.
Claims (10)
1. moxifloxacin hydrochloride injection, based on the volume of described injection, the complexing of metal ion agent of the moxifloxacin hydrochloride that comprises 1-2W/V, the solubilizing agent of 0.005-0.6W/V and 0.001-0.1W/V.
2. moxifloxacin hydrochloride injection as claimed in claim 1, wherein said solubilizing agent is selected from PLURONICS F87, Polyethylene Glycol-15-hydroxy stearic acid ester, Tween 80 or its mixture, preferably PLURONICS F87.
3. moxifloxacin hydrochloride injection as claimed in claim 1 or 2, wherein said complexing of metal ion agent is selected from sodium calcium edetate, disodium edetate, edetic acid or its mixture, preferably sodium calcium edetate.
4. the moxifloxacin hydrochloride injection as described in claim as arbitrary in claim 1-3, it also comprises pH adjusting agent.
5. moxifloxacin hydrochloride injection as claimed in claim 4, wherein said pH adjusting agent is selected from hydrochloric acid, sulphuric acid, phosphoric acid, acetic acid, malic acid, lactic acid, citric acid, tartaric acid or its mixture, preferably hydrochloric acid.
6. moxifloxacin hydrochloride injection as claimed in claim 4, wherein said pH adjusting agent is selected from sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, sodium tartrate, sodium lactate or its mixture, preferably sodium hydroxide.
7. the moxifloxacin hydrochloride injection as described in arbitrary claim in claim 4 to 6 is 4.0-5.0 at the pH value that adds described injection solution after described pH adjusting agent.
8. the method for the moxifloxacin hydrochloride injection described in arbitrary claim in preparation claim 1 to 7, it comprises:
(1) provide water for injection;
(2) add appropriate moxifloxacin hydrochloride and appropriate solubilizing agent to described water for injection;
(3a) add again appropriate complexing of metal ion agent; And
Optionally, (4a) add again pH adjusting agent so that the pH value of solution is adjusted to 4.0-5.0.
9. method as claimed in claim 8 wherein provides the water for injection of the 75%-95% of cumulative volume in step (1); The method is further comprising the steps of:
(3b) 60 ° of C-80 ° of C of insulation stirring are dissolved moxifloxacin hydrochloride and solubilizing agent completely;
(5) add water for injection to cumulative volume.
10. method as claimed in claim 8 or 9, is carrying out after described step, and described method is further comprising the steps of:
(4b) use injection remove impurity with active carbon and filter and remove active carbon; And
(6) hot pressing steam sterilizing.
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CN104771359A (en) * | 2015-04-02 | 2015-07-15 | 南京正大天晴制药有限公司 | Steady moxifloxacin hydrochloride injection |
CN105687127A (en) * | 2016-01-29 | 2016-06-22 | 上海利康瑞生物工程有限公司 | Moxifloxacin hydrochloride chitosan injection for articular cavity and preparation method thereof |
CN109260180A (en) * | 2017-07-17 | 2019-01-25 | 北京盈科瑞创新药物研究有限公司 | A kind of moxifloxacin hydrochloride Neulized inhalation pharmaceutical solutions and preparation method thereof |
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CN102000024A (en) * | 2010-11-17 | 2011-04-06 | 陕西合成药业有限公司 | Moxifloxacin hydrochloride injection, preparation method and application thereof |
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CN1663615A (en) * | 2004-03-05 | 2005-09-07 | 杨喜鸿 | Fructose injection of antibiotic medicine |
CN101836950A (en) * | 2010-03-12 | 2010-09-22 | 陕西合成药业有限公司 | Moxifloxacin hydrochloride glucose injection and preparation method and use thereof |
CN102000024A (en) * | 2010-11-17 | 2011-04-06 | 陕西合成药业有限公司 | Moxifloxacin hydrochloride injection, preparation method and application thereof |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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CN104771359A (en) * | 2015-04-02 | 2015-07-15 | 南京正大天晴制药有限公司 | Steady moxifloxacin hydrochloride injection |
CN105687127A (en) * | 2016-01-29 | 2016-06-22 | 上海利康瑞生物工程有限公司 | Moxifloxacin hydrochloride chitosan injection for articular cavity and preparation method thereof |
CN105687127B (en) * | 2016-01-29 | 2018-09-28 | 上海利康瑞生物工程有限公司 | Articular cavity moxifloxacin hydrochloride chitosan injection and preparation method thereof |
CN109260180A (en) * | 2017-07-17 | 2019-01-25 | 北京盈科瑞创新药物研究有限公司 | A kind of moxifloxacin hydrochloride Neulized inhalation pharmaceutical solutions and preparation method thereof |
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