CN104083355B - A kind of compositions for the preparation of anti-fungal keratitis eye drop - Google Patents
A kind of compositions for the preparation of anti-fungal keratitis eye drop Download PDFInfo
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Abstract
The present invention relates to a kind of compositions for the preparation of anti-fungal keratitis eye drop, it comprises the component of following weight parts: cinnamic aldehyde 1 ~ 10 weight portion, ethyl hydroxybenzoate 0.2 ~ 20 weight portion, and wherein the weight ratio of cinnamic aldehyde and ethyl hydroxybenzoate is 1: (0.2 ~ 5).Compositions for the preparation of anti-fungal keratitis eye drop provided by the invention, can be used for treating the caused keratitis such as Fusarium, Eurotium, Penicillium, Saccharomyces and corneal ulcer, the suitability is strong.Compositions for the preparation of anti-fungal keratitis eye drop provided by the invention, pass through Experiment of Compatibility, filter out cinnamic aldehyde and ethyl hydroxybenzoate compatibility uses, the bacteriostatic activity that significantly can strengthen monomer is used respectively compared to the two, be better than existing nipalgin oil preparation, there is the effect of obvious Synergistic.
Description
Technical field
The present invention relates to a kind of compositions of medicine, it can be used for the eye drop preparing treatment cornea fungal infection.
Background technology
Fungal keratitis to be a kind ofly in a bad way, to treat thorny, that blind rate is high ophthalmic infection disease.Most of patients causes perforation of cornea, blind not in time because failing to obtain early stage diagnosis and treatment or treatment, and severe patient will row eyeball excise.Since 20th century, along with increasing year by year of sickness rate, danger is also more and more paid attention to.External report, 44% central cornea ulcer is by fungus-caused, and in China, sickness rate has closely during the last ten years and significantly increases, and plant trauma is the main cause causing fungalkeratitis, and have certain seasonality and provincialism, the infected mostly is peasant.
At present, the pathogen of fungi known keratitis has kind more than 70, to China some areas as in China Guangdong, Henan, Hebei, etc. ground carry out Epidemiological study display, pathogenic bacterium are based on Fusarium, Eurotium, most area Fusarium spp. is the first pathogenic bacterium, accounting for strong position with fusarium moniliforme and Fusarinm solani, is secondly that aspergillosis, penicillium sp, candidiasis, curved spore are mould etc.
Due to less to this sick basic research at present, clinical early diagnosis and Drug therapy lack effective means, cause this sick poor prognosis, and treat thorny.Early stage diagnosis and effective Drug therapy accurately improve the key that cure rate reduces blind rate.
Ethyl hydroxybenzoate (chemical name: ethylparaben): for food, cosmetics anticorrosion, there is the antibacterial action of wide spectrum, to G
-, G
+, yeast, mycete all have good bacteriostasis." the nipalgin eye drop treatment fungal corneal ulcer 94 routine clinical observation " literary composition being published in Chinese general family medicine for 1999 discloses the nipalgin eye drop prepared with 1% ethyl hydroxybenzoate, treats fungal keratitis, ulcer is clinical obtains good curative effect.
The main chemical compositions of Cortex cinnamomi japonici (Ramulus Cinnamomi) is volatile oil, i.e. Oleum Cinnamomi, and it is a kind of volatile oil of complicated.The composition more complicated of Oleum Cinnamomi, based on cinnamic aldehyde, content is more than 80%, and other also have Cortex cinnamomi japonici (Ramulus Cinnamomi) ester, eugenol, cinnamic acid etc.Its dominant reactive part is cinnamic aldehyde.Cinnamic aldehyde clinical practice can be eased pain spasmolytic, antipyretic, strengthens blood circulation, reduces the effects such as blood glucose, antibacterial, antitumor.Its activity has bacteriostasis to various bacteria, fungus, but there is no its research report acting on fungal keratitis.
Summary of the invention
The object of the present invention is to provide a kind of formula simple, there is synergistic function, can effectively prevention and therapy fungal keratitis (Fusarium, aspergillus, Penicillium, Saccharomyces and various mycete) infect caused by fungal keratitis and corneal ulcer, and safety is good, the pharmaceutical composition for the preparation of anti-fungal keratitis eye drop had no side effect.
In order to reach the object of foregoing invention, the technical solution adopted in the present invention is:
Compositions of the present invention comprises the component of following weight parts: cinnamic aldehyde 1 ~ 10 weight portion, ethyl hydroxybenzoate 0.2 ~ 20 weight portion, and wherein the weight ratio of cinnamic aldehyde and ethyl hydroxybenzoate is 1: (0.2 ~ 5).
Further, compositions of the present invention also comprises the Tween 80 of 0.1 ~ 10 weight portion.
Further, compositions of the present invention is prepared by following method: joined by the Tween 80 of 0.1 ~ 10 weight portion in the normal saline of 900 ~ 1000 weight portions of 60 ~ 80 DEG C, add the ethyl hydroxybenzoate of 0.2 ~ 20 weight portion until completely dissolved again, and then add the cinnamic aldehyde of 1 ~ 10 weight portion, after mix homogeneously, leave standstill to room temperature both.
Further, fungus of the present invention comprises the fungus of Fusarium, aspergillus, Penicillium or Saccharomyces.
Beneficial effect of the present invention is:
Compositions for the preparation of anti-fungal keratitis eye drop provided by the invention, can be used for treating the caused keratitis such as Fusarium, Eurotium, Penicillium, Saccharomyces and corneal ulcer, the suitability is strong.
Compositions for the preparation of anti-fungal keratitis eye drop provided by the invention, pass through Experiment of Compatibility, filter out cinnamic aldehyde and ethyl hydroxybenzoate compatibility uses, the bacteriostatic activity that significantly can strengthen monomer is used respectively compared to the two, be better than existing nipalgin oil preparation, there is the effect of obvious Synergistic.
The Chinese medicine composition of anti-aspergillosis medicine provided by the invention, Tween 80 wherein uses as solubilizing agent, but the consumption of Tween 80 only has 0.01% ~ 1%(w/v), well below the consumption of 1 ~ 10% in Chinese Pharmacopoeia, the minimizing of supplementary product consumption can reduce medicine preparation cost, increases drug effect and reduces the discomfort that adjuvant causes.In prescription, the consumption of Tween 80 is 0.01% ~ 1%, and its pharmacodynamics, safety are all better than the consumption of 1 ~ 10%.
Detailed description of the invention
Embodiment 1:
Cinnamic aldehyde: 1 gram,
Ethyl hydroxybenzoate: 1 gram.
Embodiment 2:
Cinnamic aldehyde: 1 gram,
Ethyl hydroxybenzoate: 1 gram,
Tween 80: 1 gram.
Experimental example 1
Detect the activity of cinnamic aldehyde, ethyl hydroxybenzoate and the anti-fungal keratitis of Tween 80 compositions, and assembly cinnamic aldehyde, ethyl hydroxybenzoate and Tween 80 anti-fungal keratitis synergistic mensuration active to the anti-fungal keratitis of monomer cinnamic aldehyde, ethyl hydroxybenzoate, contrasts with fluconazol respectively.
Test specimen: cinnamic aldehyde >=98%, by Wuhan, Hubei, long-range Science and Technology Ltd. provides, and ethyl hydroxybenzoate is provided by Tianjin Tian Tai fine chemicals company limited, and Tween 80 is provided by Tianjin red rock chemical reagent factory, and fluconazol is provided by French Pfizer.
Medicine monomer: cinnamic aldehyde is dissolved in 95% ethanol, be mixed with 10%(w/v) mother liquid medicine, after 0.2um metre filter, be dissolved in culture medium the pharmaceutical culture medium made containing cinnamic aldehyde 1mg/ml, 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, 0.0625mg/ml, 0.3125mg/ml, 0.0156mg/ml, 0.0078mg/ml, 0.0039mg/ml respectively, each test tube subpackage 5ml places inclined-plane, and it is for subsequent use to solidify rear cold preservation.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
Ethyl hydroxybenzoate is dissolved in 95% ethanol, be mixed with 10%(w/v) mother liquid medicine, after 0.2um metre filter, be dissolved in culture medium the pharmaceutical culture medium made containing ethyl hydroxybenzoate 1mg/ml, 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, 0.0625mg/ml, 0.3125mg/ml, 0.0156mg/ml, 0.0078mg/ml, 0.0039mg/ml respectively, each test tube subpackage 5ml places inclined-plane, and it is for subsequent use to solidify rear cold preservation.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
Tween 80 is dissolved in 95% ethanol, be mixed with 10%(w/v) mother liquid medicine, after 0.2um metre filter, be dissolved in culture medium the pharmaceutical culture medium making tween 80 1mg/ml, 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, 0.0625mg/ml, 0.3125mg/ml, 0.0156mg/ml, 0.0078mg/ml, 0.0039mg/ml respectively, each test tube subpackage 5ml places inclined-plane, and it is for subsequent use to solidify rear cold preservation.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
Pharmaceutical composition: the pharmaceutical composition of embodiment 1, be dissolved in 95% ethanol respectively, be mixed with 10% mother liquid medicine, after 0.2um metre filter, be dissolved in respectively in culture medium and make A(cinnamic aldehyde 1mg/ml, ethyl hydroxybenzoate 1mg/ml), B(cinnamic aldehyde 0.5mg/ml, ethyl hydroxybenzoate 0.5mg/ml), C(cinnamic aldehyde 0.25mg/ml, ethyl hydroxybenzoate 0.25mg/ml), D(cinnamic aldehyde 0.125mg/ml, ethyl hydroxybenzoate 0.125mg/ml), E(cinnamic aldehyde 0.0625mg/ml, ethyl hydroxybenzoate 0.0625mg/ml), F(cinnamic aldehyde 0.03125mg/ml, ethyl hydroxybenzoate 0.03125mg/ml), G(cinnamic aldehyde 0.0156mg/ml, ethyl hydroxybenzoate 0.0156mg/ml), H(cinnamic aldehyde 0.0781mg/ml, ethyl hydroxybenzoate 0.0781mg/ml), I(cinnamic aldehyde 0.0039mg/ml, ethyl hydroxybenzoate 0.0039mg/ml) pharmaceutical culture medium, each test tube subpackage 5ml places inclined-plane, solidify rear cold preservation for subsequent use.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
The pharmaceutical composition of embodiment 2, is dissolved in 95% ethanol respectively, is mixed with 10% mother liquid medicine, after 0.2um metre filter, is dissolved in respectively in culture medium and makes A(cinnamic aldehyde 1mg/ml, ethyl hydroxybenzoate 1mg/ml, Tween 80 1mg/ml), B(cinnamic aldehyde 0.5mg/ml, ethyl hydroxybenzoate 0.5mg/ml, Tween 80 0.5mg/ml), C(cinnamic aldehyde 0.25mg/ml, ethyl hydroxybenzoate 0.25mg/ml, Tween 80 0.25mg/ml), D(cinnamic aldehyde 0.125mg/ml, ethyl hydroxybenzoate 0.125mg/ml Tween 80 0.125mg/ml), E(cinnamic aldehyde 0.0625mg/ml, ethyl hydroxybenzoate 0.0625mg/ml, Tween 80 0.0625mg/ml), F(cinnamic aldehyde 0.03125mg/ml, ethyl hydroxybenzoate 0.03125mg/ml, Tween 80 0.03125mg/ml), G(cinnamic aldehyde 0.0156mg/ml, ethyl hydroxybenzoate 0.0156mg/ml, Tween 80 0.0156mg/ml), H(cinnamic aldehyde 0.00781mg/ml, ethyl hydroxybenzoate 0.00781mg/ml, Tween 80 0.0078mg/ml), I(cinnamic aldehyde 0.0039mg/ml, ethyl hydroxybenzoate 0.0039mg/ml, Tween 80 0.0039mg/ml) pharmaceutical culture medium, each test tube subpackage 5ml places inclined-plane, and it is for subsequent use to solidify rear cold preservation.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
Matched group: fluconazol is dissolved in 95% ethanol, be mixed with 10%(w/v) mother liquid medicine, after 0.2um metre filter, be dissolved in culture medium the pharmaceutical culture medium made containing cinnamic aldehyde 1mg/ml, 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, 0.0625mg/ml, 0.3125mg/ml, 0.0156mg/ml, 0.0078mg/ml, 0.0039mg/ml respectively, each test tube subpackage 5ml places inclined-plane, and it is for subsequent use to solidify rear cold preservation.Wherein culture medium is the proof agar base (4% glucose, 1% peptone, 2% agar) after autoclaving.
Tested strain:
All preserve bacterial strain from No.2 Hospital, Hebei Medical Univ. Mycology Lab, be pathogenic strain (being directly from patient's corneal ulcer surface scraping secretions, the visible branched mycelia of direct microscopy, then obtain tested strain through culture identification).
Strain inoculation:
Picking has been transferred the bacterial strain of activation, and adding in 0.85% sodium chloride solution, is 1 × 10 with blood cell counting plate by bacteria suspension concentration adjustment
5 ~ 6cFU/ml.Inoculation 100ul bacteria suspension in each test tube medicine base, each concentration inoculation 2 pipe, compare with proof agar base, put in 36 DEG C of calorstats, every day observes growing state simultaneously, and record, 7 days sentence read result.
The interpretation of MIC value: by the situation of every pipe growth compared with the positive and negative control pipe, the medicine base tube grown without culture with lowest concentration of drug is for MIC terminal.
MFC measures: get glucose proteins peptone liquid base 2ml, add 2h in the test tube of bacteriostatic test asepsis growth respectively, takes out 100ul and be inoculated in sabouraud culture medium 36 DEG C and hatch 7 days, with least concentration without being grown to MFC terminal after concussion.
FIC(mark Mlc) determination: for medicine each during drug combination MIC value respectively divided by MIC value during independent medication business and.That is:
Wherein A is cinnamic aldehyde, and B is ethyl hydroxybenzoate, combines for cinnamic aldehyde and ethyl hydroxybenzoate drug combination.
Interaction is: FIC<1 has synergism, FIC >=1 but <2 has summation action, and there is antagonism FIC >=2.
Quality Control: set up the positive and negative control respectively, repeats once with program as stated above.
Statistical procedures: data acquisition SPSS13.0 software analysis.
Experimental result is in Table 1-2.
Table 1 medicine monomer corneal fungus MIC distribution situation (MIC/mgml
-1)
Table 2 pharmaceutical composition corneal fungus MFC distribution situation (MFC/mgml
-1)
From as above experimental result: the MIC value of the independent medication of cinnamic aldehyde to Fusarium spp. (fusarium moniliforme, Fusarinm solani or Fusarlum poae) is 0.0625mg/ml, with MIC value during ethyl hydroxybenzoate drug combination, fusarium moniliforme is 0.0156mg/ml, and Fusarinm solani and Fusarlum poae are 0.03125mg/ml.
The medication of cinnamic aldehyde associating ethyl hydroxybenzoate is 0.3744 to the FIC of fusarium moniliforme, to Fusarinm solani, the FIC of Fusarlum poae is 0.75, be 0.625 to the FIC of Aspergillus flavus, be 0.624 to the FIC of Aspergillus fumigatus, be 0.624 to the FIC of Candida albicans, be 1 every the FIC of bacterium mutually to chain, the FIC mould to pure green cyan is 0.3744.
Cinnamic aldehyde and ethyl hydroxybenzoate drug combination are described, the fungus for antagonism Fusarium, aspergillus, Penicillium or Saccharomyces has the effect of significant Synergistic.Cinnamic aldehyde associating ethyl hydroxybenzoate corneal fungus synergism is followed successively by: fusarium moniliforme, penicillium viridicatum, Aspergillus fumigatus, Bai Nian, Aspergillus flavus, Fusarlum poae, Fusarinm solani, has summation action every bacterium mutually to chain.
During the independent medication of Tween 80 to above-mentioned 8 kinds of funguses all without antibiotic property, do not affect the antibacterial action of cinnamic aldehyde, ethyl hydroxybenzoate during drug combination.Can be applicable to use with cinnamic aldehyde and ethyl hydroxybenzoate compatibility, the water solublity of both increases.
Fluconazol is to Candida albicans, and chain has antibacterial action every bacterium mutually, to other 6 kinds of fungal keratitis without antibacterial action.
Conclusion: the independent medication of cinnamic aldehyde, ethyl hydroxybenzoate to Fusarium spp. (beading, eggplant sick, pears spore), aspergillosis (Aspergillus flavus, Aspergillus fumigatus), Candida albicans, chain mutually every bacterium, Pure Malachitum is mould all stronger antibacterial action.Produce synergism when cinnamic aldehyde, ethyl hydroxybenzoate drug combination, strengthen further Fusarium spp. (beading, eggplant disease, pears spore), aspergillosis (Aspergillus flavus, Aspergillus fumigatus), Candida albicans, chain mutually every bacterium, the mould antibacterial action of pure green cyan.A kind of effective Therapeutic Method will be provided for treatment cornea fungal infection.
Embodiment 3
Cinnamic aldehyde: 0.5 gram,
Ethyl hydroxybenzoate: 0.1 gram,
Tween 80: 0.1 gram,
0.9% normal saline adds to 100 milliliters,
Preparation: first 0.1 gram of Tween 80 is joined in 0.9% normal saline, add ethyl hydroxybenzoate 0.1 gram successively, then add 0.5 gram of cinnamic aldehyde, enough 100 milliliters.Autoclaving, subpackage, 5 mls/branch, every day eye drip 4 ~ 6 times, each 3 ~ 5, being used in conjunction 2 weeks is a course for the treatment of.
Embodiment 4 ~ 10 and the difference of embodiment 3 are that the proportioning of each component is different, all use 0.9% normal saline to be settled to 100 milliliters, specifically in table 3.And detect the MIC value of each embodiment to fusarium moniliforme, to assess the effect that it suppresses fungal keratitis.
MIC the value (/mgml of table 3 embodiment 3 ~ 10 pairs of fusarium moniliformes
-1)
As shown in the above results, cinnamic aldehyde and ethyl hydroxybenzoate drug combination all have significant inhibitory action to fusarium moniliforme, and MIC is all less than 0.02mg/ml, are better than cinnamic aldehyde significantly or ethyl hydroxybenzoate is used alone.Be wherein 1 in the weight ratio of cinnamic aldehyde and ethyl hydroxybenzoate: time (0.2 ~ 5), the content of cinnamic aldehyde is higher, and biocidal property is more obvious.
Claims (4)
1., for the preparation of a compositions for anti-fungal keratitis eye drop, it is characterized in that it comprises the component of following weight parts:
Cinnamic aldehyde 1 ~ 10 weight portion
Ethyl hydroxybenzoate 0.2 ~ 20 weight portion
Wherein the weight ratio of cinnamic aldehyde and ethyl hydroxybenzoate is 1:(0.2 ~ 5).
2. a kind of compositions for the preparation of anti-fungal keratitis eye drop according to claim 1, is characterized in that it also comprises the Tween 80 of 0.1 ~ 10 weight portion.
3. a kind of compositions for the preparation of anti-fungal keratitis eye drop according to claim 2, it is characterized in that it is prepared by following method: joined by the Tween 80 of 0.1-10 weight portion in the normal saline of 1000 weight portions of 60-80 DEG C, add the ethyl hydroxybenzoate of 0.2-20 weight portion until completely dissolved again, and then add the cinnamic aldehyde of 1-10 weight portion, after mix homogeneously, leave standstill to room temperature and get final product.
4. a kind of compositions for the preparation of anti-fungal keratitis eye drop according to claim 3, is characterized in that described fungus comprises the fungus of Fusarium, aspergillus, Penicillium or Saccharomyces.
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Effective date of registration: 20151207 Address after: 050000 No. 215 Heping West Road, Hebei, Shijiazhuang Applicant after: No.2 Hospital, Hebei Medical Univ. Address before: 050000 No. 215 Heping West Road, Hebei, Shijiazhuang Applicant before: Wang Gangsheng |
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