CN105687127B - Articular cavity moxifloxacin hydrochloride chitosan injection and preparation method thereof - Google Patents
Articular cavity moxifloxacin hydrochloride chitosan injection and preparation method thereof Download PDFInfo
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- CN105687127B CN105687127B CN201610068700.3A CN201610068700A CN105687127B CN 105687127 B CN105687127 B CN 105687127B CN 201610068700 A CN201610068700 A CN 201610068700A CN 105687127 B CN105687127 B CN 105687127B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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Abstract
The invention belongs to pharmaceutical technology fields, and in particular to a kind of articular cavity moxifloxacin hydrochloride chitosan injection and preparation method thereof comprising following components：The moxifloxacin hydrochloride of 0.3 0.8wt%, the chitosan of 0.1 0.8wt%, the surfactant of 0.005 0.02wt%, isotonic regulator and pH adjusting agent, surplus is water for injection, the addition of isotonic regulator be make the osmotic pressure of composition be 250 350 millisomoles/liter, the addition of pH adjusting agent is that the pH of composition is made to be 6.5 7.5.Articular cavity prepared by present invention moxifloxacin hydrochloride chitosan injection, can be effectively relieved pain caused by simultaneously treatment of arthritis, and therapeutic effect is notable.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to a kind of articular cavity moxifloxacin hydrochloride chitosan injection and
Preparation method.
Background technology
Osteoarthritis is a kind of using Articular cartilage degeneration and secondary osteoproliferation as the chronic joint diseases of characteristic.
It is more common in the middle-aged and the old, women is more than male.It is mainly in the larger knee joint of bearing a heavy burden, hip joint, lumbosacral region joint of vertebral column and the
Distal interphalangeal joint, the proximal interphalangeal joint of the positions such as one Plantar toe joints and hand.The disease also becomes osteoarthropathy, regression
Property arthritis, hyperplasia row arthritis, degenerative arthritis, Osteoarthritis, infectional arthritis etc..With the aging of population with
And fat ratio rises, the prevalence rate of osteoarthritis will improve.
Osteoarthritis can fall ill since 20 years old, but most of asymptomatic, generally be not easy to find.The World Health Organization unites
It counting, in 50 years old or more crowd, the incidence of osteoarthritis is 50%, in 55 years old or more crowd, incidence 80%.China's bone
Arthritic incidence accounts for about the 10% of total population, is 100,000,000 people or so.Nineteen ninety, China only have ten thousand osteoarthritis more than 4000
Patient, and 80,000,000 are had reached within 2000, patient numbers have reached people more than 100,000,000, and China will become world osteoarthritis illness people
Most one of the country of number.
The cause of disease of osteoarthritis is the synovia (joint fluid) due to having lacked viscosity in articular cavity, causes to serve as originally
As the abnormal friction of the cartilage of cushion in Bones and joints, damages and degenerate.After cartilage degradation, bone table just can not be protected
Face, in walking or standing, body wt makes the joint degenerated more painful.Due to fearing pain, reduce movement naturally,
Then the muscle of there also follows atrophy, and ligament can also become more loose.
The main pathological change of osteoarthritis be at cartilage degeneration and disappearance and joint margins ligament attachment and
Subchondral bone qualitative response hyperplasia forms spur, and thus causes arthralgia, stiff deformity and dysfunction.The disease is in clinic
On, primary and secondary two class can be divided into.Spontaneous osteoarthritis means age ageing and the relevant pass of other diseases of getting along well
Lesion is saved, secondary osteoarthritis is then caused by the diseases such as damage, inflammation, heredity and metabolism, endocrine.
Infectional arthritis is one kind of arthritis, refers to caused in the microorganisms such as bacterium, virus invasion articular cavity
Arthritis.Patient is mostly the weaker children and the elderly of passive protective physical fitness.The infection of joint is most common the reason is that septicemia, is removed
Except this, wound, operation, juxtra-articular soft tissue infection, and morbidity major reason.Infectional arthritis is according to the cause of disease
It is divided into two classes：
1. acute infective arthritis
It can be caused by bacterium or virus infection：(1) Neisseria gonorrhoeae arthritis, Neisser is gonococcal to be used as pathogen, it
Some hand Minor articulus, elbow, knee joint and ankle-joint are expanded to from infection mucous membrane surface (cervix, rectum, pharynx), axial skeleton closes
Save less involve；(2) Nongonococcal arthritis, mostly by staphylococcus aureus, streptococcus and gram-negative bacteria, such as intestines bar
Bacterium, Pseudomonas aeruginosa, serratia marcecens cause；(3) staphylococcus aureus and the infection of B group of streptococcus, are more common in newborn and 2 one full year of life
Above children；(4) infection of joint anaerobic bacteria is often accompanied by facultative or aerobic bacteria infection (5%~10%), such as Staphylococcus aureus
Bacterium, epidermis streptococcus and Escherichia coli；(5) the caused infection of joint after being bitten, mostly by gram-negative bacteria such as Type B streptococcus,
Oral cavity anaerobium (such as clostruidium, streptococcus, bacteroid) causes.The caused infection of joint is often golden yellow after animal bite
Color staphylococcus or mouth-flora；(6) infection of joint in HIV patient, by staphylococcus aureus, streptococcus, detection of Salmonella causes,
HIV patient can have reiter syndrome, relapsing arthritis, HIV correlations arthritis and arthralgia.HIV patient's survival is more long,
The infection chance of mycobacteria, fungi and rare conditioned pathogen is also more.
2. chronic infectional arthritis
Chornic arthritis can be caused by mycobacteria, fungi and some other pathogenic weaker bacterium.Such as tuberculosis branch
Bacillus, Mycobacterium marinum, mycobacterium kansasii, Mycotoruloides, coccidioides immitis category, Histoplasma capsulatum, Novel hidden
Coccus, the raw bacterium of dermatitis tooth, Sporothrix schenckii Pseudomonas, Aspergillus, actinomyces israelii and Brucella.There is 2/3 patient articular to set
It changes postoperative infection to be happened within 1 year, this may be since surgical procedure introduces bacterium or such as skin sense of postoperative bacterium infection
Dye, pneumonia, dentistry infection etc..
Osteoarthritis is the performance of Bones and joints physiology sexual involution, there is no reverse or stops the drug of the disease progression.Treatment
Purpose is to mitigate pain, alleviates symptom, prevents and delay the development of disease, Saving cortilage function, to prevent maimed person.Using Synthetic
It treats, including patient education, drug therapy, physical therapy and surgical operation therapy.
The drug therapy of current common osteoarthritis has following four kinds：
(1) improve the drug of symptom：Analgestic such as paracetamol has analgesic activity, but anti-inflammatory effect is weak.Non-steroidal
Anti-inflammatory agent has the characteristics of anti-inflammatory analgesic, can mitigate arthralgia after medication, improves range of motion.
(2) glucocorticoid：Unsuitable systemic administration, only invalid to other treatment, there is acute inflammation breaking-out performance in joint
Or having joint surrounding synovial membrane inflammation, skin inflammation etc. can give in articular cavity or diseased region local injection, unsuitable Reusability.It is same
Position biphasic injection interval time is at least at 3 months or more.
(3) Chondroprotective agents are used：Symptom can be alleviated, maintain and restore function of joint, such as poly glucosamine.
(4) viscosupplementation：It is Sodium Hyaluronate, the chitin sugar juice to intraarticular injection macromolecule, subtracts
Light synovial membrane inflammation, cartilage destruction and improvement function of joint, block the vicious circle of local patholoic change.On viscoplasticity object replacement therapy is
The medicine new concept that century 70 proposes, i.e. intraarticular injection have the one of viscoelastic Substance treatment osteoarthritis
The clinical new method of kind.Visco-elastic material used is mainly Sodium Hyaluronate, chitosan etc. so far.
The main component of chitosan is aminoglucan, has good biocompatibility and biodegradability, in body
It is interior it is degradable be aminoglucose, aminoglucose be articular cartilage in basis, human chondrocytes synthetic proteins can be stimulated
Polysaccharide can supplement knuckle synovia.In addition, chitosan has height viscoplasticity, and it is similar to normal knuckle synovia, it can simulate
The physical action of knuckle synovia improves the lubrication state in joint, and mitigation can be covered in soft in the pressure of articular cartilage face
Bone surface is filled into the cartilage crack of regression, and it is thin to prevent harmfulness cell factor and cartilage matrix and cartilage in knuckle synovia
Born of the same parents contact, and prevention effect is played to cartilage degeneration.
As a kind of visco-elastic material of intraarticular injection, chitosan has following performance：(1) chitosan has selection
Property promote epithelial cell, endothelial cell growth and inhibit the biological nature of fibroblastic growth, to promote tissue physiology's property
It repairs, scar is inhibited to be formed, reduce tissue adhesion；(2) chitosan has local hemostasis effect and inhibits fibrin beam shape
At organizing adhesion caused by reduce due to organization of hematoma；(3) chitosan has lubricating action and biological barrier effect, energy
Effectively prevent adhesion.The mechanism of chitosan Saving cortilage cartilage be chitosan physics and chemistry in nature with intra-articular amino
Polysaccharide is similar, has viscoplasticity, delays absorbability, and cartilage and cartilage matrix constitute the basis with metabolism when glycosaminoglycan, display
Higher than natural joint liquid.
Moxifloxacin (Avelox, Avalox) is forth generation fluoroquinolone antibiotics, and Yuan Yan producers are Bayer Bitterfeld GmbH
Company, three generations's quinolone drugs is wider earlier above for antibiotic, and trade name " visits multiple pleasure ", is listed in Germany in September, 1999, together
December in year obtains FDA approval listings in the U.S., and the market sales revenue of 2002 " visiing multiple pleasure " is more than 300,000,000 dollars, and it is big to become the world ten
One of situation of selling well antibiotic.The product of Bayer A.G and Schering Plough company of the U.S. is up to for 2006 in world's market sales revenue
800000000 dollars, in global situation of selling well prescription medicine ranking 129；Its market sales revenue was up to 10.34 hundred million dollars within 2007, compared with 2006
Increase by 25.8%；Its sales volume in 2008 is more than 1,100,000,000 dollars.2002, Moxifloxacin piece listed in China, by Bayer public affairs
Department is sold, and key market is the main hospital of China big and medium-sized cities.The medicine enters national medical insurance directory for 2004, hereafter
Surprising growth rate is presented within 3 years；Compound growth rate to Moxifloxacin in 2007 in 2003 is 116%, city sample in 2007
Hospital's money for drugs surpasses 2.16 hundred million yuan, increases by 75.1% compared with 2006；Sales volume in China in 2008 surpasses 300,000,000 yuan.
Invention content
Invention broadly provides a kind of articular cavity moxifloxacin hydrochloride chitosan injections and preparation method thereof, can
Pain caused by simultaneously treatment of arthritis is effectively relieved, therapeutic effect is notable.Its technical solution is as follows：A kind of articular cavity with hydrochloric acid not
Xisha star chitosan injection comprising following components：The moxifloxacin hydrochloride of 0.3-0.8wt%, the chitin of 0.1-0.8wt%
Sugar, the surfactant of 0.005-0.02wt%, isotonic regulator and pH adjusting agent, surplus are water for injection, isotonic regulator
Addition be make the osmotic pressure of composition be 250-350 millisomoles/liter, the addition of pH adjusting agent is to make the pH of composition be
6.5-7.5。
Preferably, articular cavity moxifloxacin hydrochloride chitosan injection comprising following components：0.5wt%'s
Moxifloxacin hydrochloride, the chitosan of 0.3wt%, the surfactant of 0.01wt%, isotonic regulator and pH adjusting agent, surplus are
Water for injection, the addition of isotonic regulator be make the osmotic pressure of composition be 260-320 millisomoles/liter, pH adjusting agent plus
It is that the pH of composition is made to be 6.8-7.2 to enter amount.
Preferably, the isotonic regulator is sodium chloride.
Preferably, the surfactant is Tween-80.
Preferably, the pH adjusting agent is boric acid.
A kind of articular cavity preparation method of moxifloxacin hydrochloride chitosan injection, includes the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) pH adjusting agent of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and isotonic regulator are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) it is moxifloxacin hydrochloride chitosan injection to use poly (ether sulfone) film filtration sterilization, acquired solution.
Preferably, the aperture of poly (ether sulfone) film filtration sterilization is 0.22-0.45 μm in step (5).
Using above-mentioned articular cavity moxifloxacin hydrochloride chitosan injection and preparation method thereof, the present invention has following excellent
Point：
Moxifloxacin treats infectional arthritis, and mainly playing potent antibacterial to acute and chronic arthritis caused by bacterium disappears
Scorching effect.Cooperation chitosan is injected into articular cavity, can play the effect of anti-inflammatory is with analgesic.The injection of the present invention uses
Moxifloxacin hydrochloride and chitosan, stability is good, curative effect is good, resistant rate is low, adverse reaction is few, can be effectively relieved and treat pass
Pain caused by section is scorching, therapeutic effect are notable.
Specific implementation mode
1, formula of liquid is injected
The surface work of the moxifloxacin hydrochloride of 0.3-0.8wt%, the chitosan of 0.1-0.8wt%, 0.005-0.02wt%
Property agent, isotonic regulator and pH adjusting agent, surplus is water for injection, and the addition of isotonic regulator is to make the osmotic pressure of composition
For 250-350 millisomoles/liter, the addition of pH adjusting agent is that the pH of composition is made to be 6.5-7.5.
2, injection preparation
Include the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) pH adjusting agent of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and isotonic regulator are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) it is moxifloxacin hydrochloride chitosan injection to use poly (ether sulfone) film filtration sterilization, acquired solution.
One, specific embodiment
Embodiment 1
1, formula of liquid is injected
The moxifloxacin hydrochloride of 0.5wt%, the chitosan of 0.3wt%, the Tween-80 of 0.01wt%, sodium chloride and boric acid,
Surplus is water for injection, the addition of sodium chloride be make the osmotic pressure of composition be 260 millisomoles/liter, the addition of boric acid is
It is 6.8 to make the pH of composition.
2, injection preparation
Include the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) boric acid of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and sodium chloride are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) use aperture for 0.22 μm of poly (ether sulfone) film filtration sterilization, acquired solution is moxifloxacin hydrochloride chitosan note
Penetrate liquid.
Embodiment 2
1, formula of liquid is injected
The moxifloxacin hydrochloride of 0.8wt%, the chitosan of 0.1wt%, the Tween-80 of 0.02wt%, sodium chloride and boric acid,
Surplus is water for injection, the addition of sodium chloride be make the osmotic pressure of composition be 320 millisomoles/liter, the addition of boric acid is
It is 7.2 to make the pH of composition.
2, injection preparation
Include the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) boric acid of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and sodium chloride are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) use aperture for 0.45 μm of poly (ether sulfone) film filtration sterilization, acquired solution is moxifloxacin hydrochloride chitosan note
Penetrate liquid.
Embodiment 3
1, formula of liquid is injected
The moxifloxacin hydrochloride of 0.3wt%, the chitosan of 0.8wt%, the Tween-80 of 0.005wt%, sodium chloride and boron
Acid, surplus are water for injection, the addition of sodium chloride be make the osmotic pressure of composition be 320 millisomoles/liter, the addition of boric acid
To make the pH of composition be 7.5.
2, injection preparation
Include the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) boric acid of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and sodium chloride are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) use aperture for 0.35 μm of poly (ether sulfone) film filtration sterilization, acquired solution is moxifloxacin hydrochloride chitosan note
Penetrate liquid.
Two, auxiliary material Study on Compatibility
For research moxifloxacin hydrochloride, chitosan and auxiliary material consistency problem, following experiment is done：
Experiment 1：Moxifloxacin hydrochloride 0.5g, chitosan 2g, injection water 100mL；
Experiment 2：Moxifloxacin hydrochloride 0.5g, chitosan 2g, boric acid 0.6g, injection water 100mL；
Experiment 3：Moxifloxacin hydrochloride 0.5g, chitosan 2g, sodium chloride 0.5g, injection water 100mL；
Experiment 4：Moxifloxacin hydrochloride 0.5g, chitosan 2g, boric acid 0.6g, sodium chloride 0.5g, injection water 100mL.
Above-mentioned prescription is prepared and measures related substance after completing, is described in table 1 below.
1 related substances measurement result of table
As shown in Table 1, related substances difference under influence factor is little in experiment 1-4, illustrates auxiliary material to related substance shadow
It rings less, auxiliary material compatibility is good.
Three, Experiment of Zoology
This experiment mainly studies articular cavity with moxifloxacin hydrochloride chitosan injection to traumatic arthritis Rabbit Model
Observation of curative effect is as follows：
(1) 8~15 2.0~3.0kg of monthly age (♀) rabbit 40 are chosen, 30 are served only for modeling, and 10 are normal control；
(2) after anaesthetizing the rabbit of 30 modelings with yellow Jackets, knee joint disinfection on the right side of animal is selected, by document
Method carries out meniscectomy, and then by skin closure, the same day proceeds by penicillin intramuscular injection, totally 7 days；
(3) rabbit of 30 modelings is randomly divided into tri- groups of A, B, C, every group 10, A groups rabbit is prepared with embodiment 1
Injection gives each 2ml treatments, and injection in 7 days is primary, injects within continuous 5 weeks；B groups rabbit is given every time with chitosan injection
2ml is treated, and injection in 7 days is primary, is injected within continuous 5 weeks；C groups are not treated；10 control groups are D groups, the rabbits of D groups normally not by
Wound；
(4) TNF-α content in an animal blood Samples detection blood sample is respectively taken after postoperative 12,14,16 weeks.By TNF-α mark
Quasi- product add buffer solution 1mL, and it is respectively 0.3,0.9,2.7,8.1,24.3 μ g/L to adjust concentration, and 100 μ L is respectively taken to be added in test tube, then
It sequentially adds in 100 μ L to standard pipe of 125I-TNF and anti-tnf-alpha serum, is placed for 24 hours for 40 DEG C after mixing well, add PR points
From 500 μ L mixings of agent, 20min, 4 DEG C of centrifugations are placed, 3500r/min centrifuges 25min.Supernatant is abandoned in suction, on automatic gamma counter
Precipitation number is measured, standard curve is drawn and calculates sample concentration, statistical variance analyzes group difference.
(5) result：Four groups of rabbit articular conditions are C group ＜ B group ＜ A group ＜ D groups.12 weeks after modeling, A groups, B groups, C group blood
Slurry and joint irrigation TNF-α content are obviously higher than D groups (p ＜ 0.01), and the TNF- of 12,14,16,18 weeks Each point in time
Alpha content, C groups are significantly higher than B groups, A groups, D groups, and the TNF-α content at A groups each time point is low compared with B groups (p ＜ 0.05), is shown in Table 2.
TNF-α content results in 2 rabbit different time points blood plasma of table, joint fluid
It follows that moxifloxacin hydrochloride chitosan injection prepared by the present invention has well infectional arthritis
Healing acts on.
It will be apparent to those skilled in the art that technical solution that can be as described above and design, make various other
Corresponding change and deformation, and all these changes and deformation should all belong to the protection domain of the claims in the present invention
Within.
Claims (6)
1. a kind of articular cavity moxifloxacin hydrochloride chitosan injection comprising following components：The hydrochloric acid of 0.3-0.8wt% is not
Xisha star, the chitosan of 0.1-0.8wt%, the surfactant of 0.005-0.02wt%, isotonic regulator and pH adjusting agent, it is remaining
Amount be water for injection, the addition of isotonic regulator be make the osmotic pressure of composition be 250-350 millisomoles/liter, pH adjusting agent
Addition be that the pH of composition is made to be 6.5-7.5, surfactant is Tween-80.
2. articular cavity according to claim 1 moxifloxacin hydrochloride chitosan injection comprising following components：
The moxifloxacin hydrochloride of 0.5wt%, the chitosan of 0.3wt%, the surfactant of 0.01wt%, isotonic regulator and pH are adjusted
Agent, surplus are water for injection, the addition of isotonic regulator be make the osmotic pressure of composition be 260-320 millisomoles/liter, pH tune
The addition for saving agent is that the pH of composition is made to be 6.8-7.2.
3. articular cavity according to claim 1 moxifloxacin hydrochloride chitosan injection, it is characterised in that：Isotonic adjusting
Agent is sodium chloride.
4. according to claim 1-3 any one of them articular cavity moxifloxacin hydrochloride chitosan injection, it is characterised in that：
PH adjusting agent is boric acid.
5. a kind of articular cavity as described in claim 1 preparation method of moxifloxacin hydrochloride chitosan injection, feature
It is：Include the following steps：
(1) chitosan for weighing formula ratio is dissolved in the water for injection of 95% volume, is stirred；
(2) pH adjusting agent of formula ratio is added in chitin sugar juice, is stirred evenly；
(3) moxifloxacin hydrochloride and isotonic regulator are added into the solution of step (2)；
(4) water for injection adds to full dose, stirs evenly；
(5) it is moxifloxacin hydrochloride chitosan injection to use poly (ether sulfone) film filtration sterilization, acquired solution.
6. the articular cavity according to claim 5 preparation method of moxifloxacin hydrochloride chitosan injection, feature exist
In：The aperture of poly (ether sulfone) film filtration sterilization is 0.22-0.45 μm in step (5).
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| CN105687127A (en) | 2016-06-22 |
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