CN102349926A - New medical application of combination of paeonol, derivative thereof and paeoniflorin - Google Patents
New medical application of combination of paeonol, derivative thereof and paeoniflorin Download PDFInfo
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Abstract
The invention discloses an application of the combination of paeonol, a derivative thereof and paeoniflorin in preparation of a drug for treating eye diseases. The invention explores a new medical application of the combination of the known compound paeonol, the derivative thereof and the paeoniflorin, namely the application of the combination of the paeonol, the derivative thereof and the paeoniflorin in preparation of the drug for treating eye diseases, and opens up a new application area. The combination of the paeonol, the derivative thereof and the paeoniflorin can be used in combination modes with different proportions and concentration and has obvious synergy effect. The invention is good for realizing quality controllability and repeatability of Chinese herbal preparation as well as the in-depth study of the pharmacological action and the mechanism of action of the Chinese herbal preparation, and is good for the development and application of innovative Chinese herbal medicine. The product provided by the invention has rich raw material source, the raw materials are cheap, and the paeonol, paeonol sodium and the paeoniflorin are safe and low-toxic and have strong pharmacological effects, which indicates a good medical application prospect.
Description
Technical field
The present invention relates to medical technical field, relate in particular to the new medical use of paeonol and derivant thereof and peoniflorin combination, be specifically related to paeonol and derivant thereof and peoniflorin and be combined in the purposes in the preparation treatment oculopathy medicine.
Background technology
Existing document discloses compound paeonolum (Paeonol) and part derivant thereof, and the structural formula of paeonol is:
Chemical formula is: 2-hydroxyl-4-methoxyacetophenone, molecular formula are C
9H
10O
3, relative molecular mass is 166.18.English by name: Paeonolum.Another name: peonol, paeonol.This chemical compound has the characteristic of fusing point low [mp (51+1) ℃].
One of its derivant sodium paeonol sulfonate be paeonol after sulfonation and obtain, chemical constitution is 2-methoxyl group-4 hydroxyls-5-acetylbenzene sodium sulfonate, is characterized in that its water solublity is significantly improved, its pharmacological action and paeonol are basic identical.
2-methoxyl group-4-hydroxyl-5-acetyl group diphenyl diimide-4-arsenic acid is that paeonol and atoxylic acid carry out the derivant that coupling reaction makes, and has been characterized in strengthening the pharmacological action of paeonol, and its pharmacological action and paeonol are basic identical.
Paeonol can make by extracting in Chinese medicine Cortex Moutan [root bark of ranunculaceae peony (Paeonia suffruticosa)] and the Chinese medicine Radix Cynanchi Paniculati [herb of Asclepiadaceae plant Radix Cynanchi Paniculati (Cynanchum paniculatum)]; But also chemosynthesis; The known active component that can be used as a treatment seat skin ulcer, skin-whitening, cardiovascular disease, rheumatic arthritis, antineoplastic agent, Chinese scholars confirms that through years of researches paeonol is the topmost drug activity composition of Cortex Moutan in recent years.The commodity of at present existing paeonol and paeonol sodium sulfonate.
Existing document also discloses chemical compound peoniflorin (Paeoniflorin), and the structural formula of peoniflorin is:
The molecular formula of peoniflorin is: C
23H
28O
11, relative molecular mass is 480.45.English by name: Paeoniflorin.Peoniflorin can derive from the root of ranunculaceae plant Radix Paeoniae Paeonia albiflora Pall, the root of Paeonia suffruticosa P.suffrsticosa Andr; The root of Paeonia delavayi P.delarayi Franch; Content with Radix Paeoniae Rubra be high [Wang Yuntao, Wang Limei, Jin Xiangqun. the high-efficiency liquid-phase fingerprint of Radix Paeoniae Rubra and hygroplasm combination analysis. Chinese patent medicine; 2010,32 (7): 1089-1092.Wang Yanzhi, Shi Renbing, Liu Bin. the separation of Radix Paeoniae Rubra chemical constituent and structure are identified. Chemistry for Chinese Traditional Medicine, 2006,29 (4): 267-269].Though motherland's medical science is divided into the Radix Paeoniae Alba and Radix Paeoniae Rubra with Radix Paeoniae, it is generally acknowledged that the root convection drying of the wild article of Radix Paeoniae then is Radix Paeoniae Rubra, and be the Radix Paeoniae Alba behind the root peeling decocting in water of cultivation article.All contain monoterpenes chemical compounds such as peoniflorin, lactone glucoside of Radix Paeoniae, Hydroxy peoniflorin, benzoylpaeoniflorin in white peony root's total glycoside and the total paeony glycoside, and peoniflorin is the characteristic component of the Radix Paeoniae Alba and Radix Paeoniae Rubra, but the amount of the peoniflorin in the Radix Paeoniae Rubra is far above the Radix Paeoniae Alba.[Chinese Pharmacopoeia 2000 editions, one one (S) published the 78th page, the 125th page in 2000.Brilliant, Zhao Lei, the Zhao Guang tree Radix Paeoniae Alba and the comparative study of Radix Paeoniae Rubra HPLC finger printing, Chinese herbal medicine, 2010,41 (11): 1904-1906].Modern pharmacological research shows: effect such as Radix Paeoniae has immunomodulating, analgesia, spasmolytic, protects the liver, blood vessel dilating, antiinflammatory; Peoniflorin is the main drug activity composition [Zhou Qiang of then Radix Paeoniae; Foxtail millet accounts for state. the pharmacological action of Radix Paeoniae Alba total glucosides and the application in autoimmune disease thereof. and Chinese Journal of New Drugs and Clinical Remedies; 2003,22 (11): 687-690.Jiang Jianping. modern pharmacological research of the Radix Paeoniae Alba and clinical practice overview. Chinese medicine information, 2000, (3): 6-8].The medicine of the commodity of at present existing peoniflorin and peoniflorin injection, white peony root's total glycoside (wherein, content of paeoniflorin >=90%).
In the classic doctor of successive dynasties traditional Chinese medical science nationality; We can see a plurality of classics sides of containing Cortex Moutan and Radix Paeoniae compatibility treatment disease; Be used for epidemic febrile disease heat at blood system like what put down in writing in " prescriptions worth thousand gold "; Fever of the body, a macule, and spit blood due to the heat in blood and wait the xijiao Dihuang Tang (Cornu rhinocerotis, Radix Rehmanniae, Cortex Moutan, Radix Paeoniae Rubra) of card; Ramulus Cinnamomi Poria pill that is used for cards such as amenorrhea due to stagnation of blood, dysmenorrhea ， Huo mass in the abdomen of record in " Medical Treasures of the Golden Chamber " (Ramulus Cinnamomi, Poria, pink, Cortex Moutan, Radix Paeoniae); Record is used for moving back of hyperemia of the bulbar conjunctiva red loose (Cortex Moutan, Radix Paeoniae Rubra, Cortex Mori, Radix Scutellariae, Radix Trichosanthis, Radix Platycodonis, Radix Angelicae Sinensis, Radix Ophiopogonis, Semen Trichosanthis, Radix Glycyrrhizae) in the TCM Ophthalmology monograph " Precious Book of Ophthalmology "; In " prescription of peaceful benevolent dispensary " record be used for married woman's heat in blood deficiency of vital energy taste blood soup (Cortex Moutan, Radix Paeoniae Rubra, Radix Angelicae Sinensis, Rhizoma Chuanxiong, Herba Schizonepetae, Herba Verbenae, Cortex Cinnamomi, Fructus Aurantii); The JIAWEI XIAOYAO SAN (Cortex Moutan, the Radix Paeoniae Alba, Fructus Gardeniae, Radix Bupleuri, Radix Angelicae Sinensis, Poria, the Rhizoma Atractylodis Macrocephalae, Radix Glycyrrhizae, Rhizoma Zingiberis Recens, Herba Menthae) of the card of putting down in writing in " women section brief points " that is used for stagnation of QI due to depression of the liver etc.Cortex Moutan and Radix Paeoniae Rubra are just paid attention in explanation in traditional tcm prescription compatibility compatibility uses.Theoretical according to traditional Chinese medicine, Cortex Moutan and Radix Paeoniae Rubra compatibility have the mutual reinforcement between effect.
Which type of pharmacological effect paeonol and derivant thereof in forming different prescriptions, different ratio reorganization can be produced with peoniflorin? The research of this respect mainly concentrates on the influence to cardiovascular system of paeonol and peoniflorin prescription at present.Like Guo Qi; Deng; The influence acute of Cortex Moutan effective ingredient reorganization compound recipe to the dog Experimental Acute Myocardial Infarction; China's Chinese medicine information magazine, has reported that each compatibility group of paeonol and peoniflorin all is superior to Cortex Moutan decocting liquid group in the effect that alleviates degree of ischemia and reduction ischemia scope, dwindle aspect the myocardial infarction area at 2010 17 the 4th phases of volume.Li Yikui, etc. the protective effect to rats with acute myocardial infarction of, paeonol and peoniflorin and different proportioning thereof, new Chinese medicine and clinical pharmacology, have been reported the peoniflorin (25mgkg of various dose at 2010 21 the 3rd phases of volume
-1, 50mgkg
-1), the paeonol (50mgkg of various dose
-1, 100mgkg
-1) and the different proportioning (50mgkg of paeonol and peoniflorin
-1: 25mgkg
-1, 50mgkg
-1: 50mgkg
-1, 100mgkg
-1: 50mgkg
-1, 100mgkg
-1: 25mgkg
-1) to myocardial infarction area, ECG ST section and creatine kinase, the TB of rats with acute myocardial infarction
2Effect, the result shows paeonol 100mgkg
-1Group and contain paeonol 100mgkg
-1Two compatibility groups, paeonol 50mgkg
-1+ peoniflorin 50mgkg
-1Group, paeonol 50mgkg
-1Group all can obviously be improved each experimentation index, and significant improvement effect does not all appear in single two groups with peoniflorin.
Because natural medicinal ingredients is complicated, the different pharmaceutical compatibility uses possibly produce different effects; The reorganization compound recipe of different pharmaceutical effective ingredient also possibly produce different effects.Which type of with Cortex Moutan and Radix Paeoniae two herbal medicines combination, especially its combination of active principles, be difficult to expect in performance effect aspect prevention or the treatment oculopathy.Still do not have at present is the pertinent literature report or the patent of material combination treatment oculopathy with Cortex Moutan and Radix Paeoniae two flavors only; Do not have yet will be wherein the active component extract compatibility or wherein topmost active ingredient paeonol and peoniflorin compatibility and pharmaceutically acceptable one or more carriers or adjuvant or the complementary composition pharmaceutical composition or the preparation that combine to process be used to treat or prevent oculopathy, the relevant report of oculopathy such as especially immune-related conjunctivitis, chronic conjunctivitis, scleritis, uveitis.
Summary of the invention
The invention discloses paeonol and derivant thereof and peoniflorin and be combined in the purposes in the preparation treatment oculopathy medicine.The present invention also provides this preparation of drug combination method.
In fact; Oculopathy such as immunity conjunctivitis, uveitis, chronic conjunctivitis, xerophthalmia, scleritis still do not have ideal Therapeutic Method at present, and medical circle is still at the new medicine that strives to find oculopathy such as the littler treatment immunity conjunctivitis of better efficacy, side effect, uveitis, chronic conjunctivitis, xerophthalmia, scleritis.The present invention is through research repeatedly, and accomplished the present invention through the repeated validation of animal and clinical trial.
The invention reside in the new purposes that paeonol and derivant and peoniflorin combination are provided, i.e. new application in pharmacy.
In fact, the present invention relates to paeonol and derivant thereof and peoniflorin and be combined in the application in the medicine for preparing treatment oculopathy.
Relate to paeonol and derivant thereof and peoniflorin and be combined in the application in the medicine for preparing treatment immunity conjunctivitis.Described immunity conjunctivitis is called allergic conjunctivitis again, mainly comprises anaphylaxis conjunctivitis, long-term anaphylaxis conjunctivitis, seasonal allergic conjunctivitis, vernal keratoconjunctivitis, macropapillary conjunctivitis, eczematous keratoconjunctivitis, autoimmune membranous conjunctivitis etc.
Relate to paeonol and derivant thereof and peoniflorin and be combined in the application in the medicine for preparing the treatment xerophthalmia.Xerophthalmia is claimed xerophthalmia again.
Relate to paeonol and derivant thereof and peoniflorin and be combined in the application in the medicine for preparing the treatment chronic conjunctivitis.
Also relate to paeonol and derivant thereof and peoniflorin and be combined in the application in the uveitic medicine of preparation treatment.
Also relate to paeonol and derivant thereof and peoniflorin and be combined in the application in the medicine for preparing treatment scleritis.
Above-mentioned paeonol and derivant thereof comprise the series derivates of paeonol and analog structure thereof.The series derivates of said paeonol analog structure can be to have similar chemical constitution by what extraction in natural material such as the plant obtained with paeonol, particularly has similar pharmacological action derivant with paeonol.The series derivates of said paeonol analog structure also can be to be the derivant of raw material through chemosynthesis with the paeonol.
In above-mentioned paeonol and derivant thereof and the peoniflorin combination, peoniflorin can be used with the chemical compound of peoniflorin analog structure or the derivant of peoniflorin and substitute, like Hydroxy peoniflorin, benzoylpaeoniflorin etc.The chemical compound of said peoniflorin analog structure or the derivant of peoniflorin can be to obtain by extracting in natural material such as the plant, also can be that raw material is through chemosynthesis with the peoniflorin.The chemical compound of peoniflorin analog structure or derivant have identical or similar pharmacology and drug action with peoniflorin, and pharmacological action and peoniflorin are basic identical in the combined preparation medicine of above-mentioned and paeonol and derivant thereof.
Combinations thereof comprises: the series derivates of paeonol, paeonol analog structure or paeonol and derivant thereof are no less than series derivates or peoniflorin and the combination that derivant is no less than two kinds mixture thereof of two kinds mixture and peoniflorin, peoniflorin analog structure; As, paeonol and peoniflorin combination, a kind of combination of paeonol and peoniflorin analog structure series derivates, paeonol and peoniflorin and derivant thereof are no less than the combination of two kinds mixture; As: a kind of and peoniflorin combination of the series derivates of paeonol analog structure; A kind of combination of a kind of and peoniflorin analog structure series derivates of the series derivates of paeonol analog structure, a kind of and peoniflorin of the series derivates of paeonol analog structure and derivant thereof are no less than the combination of two kinds mixture; As: paeonol and derivant thereof are no less than two kinds mixture and peoniflorin combination, as above analogize.
Above-mentioned paeonol and derivant thereof and peoniflorin make up by weight, and the ratio of mark is 0.5: 1 to 4: 1; It is paeonol and derivant thereof: peoniflorin=0.5: 1 to 4: 1.
The combination of above-mentioned paeonol and derivant thereof and peoniflorin can be prepared from pharmaceutical composition or the preparation that paeonol and derivant thereof and peoniflorin make up with pharmaceutically acceptable one or more carriers or adjuvant or complementary composition, and the weight percentage that paeonol and derivant thereof and peoniflorin are combined in pharmaceutical composition or the preparation is 0.02% to 99%.Said weight percentage is 0.02% to 99% to be meant that mixture that paeonol and derivant thereof and peoniflorin combination form is prepared from pharmaceutical composition that paeonol and derivant thereof and peoniflorin make up or the weight percentage in the preparation adding other pharmaceutically acceptable one or more carriers or adjuvant or complementary composition.
The combination of above-mentioned paeonol and derivant thereof and peoniflorin can be prepared into eye with liquid preparation, eye with semi-solid preparation, eye with solid preparation, with sustained release pharmaceutical formulation, with film controlled release medicine preparation, sclera release preparation, capsule, pill, tablet, granule, injection, powder etc. are used to treat and prevent the medicine of oculopathy thoroughly.
Above-mentioned paeonol and derivant thereof become weight percentage that eye makes up with paeonol in the liquid preparation and derivant thereof and peoniflorin more than or equal to 0.02% with the peoniflorin combined preparation.
According to Pharmacopoeia of the People's Republic of China version in 2005, above-mentioned eye is meant eye drop, collyrium, intraocular injection agent with liquid preparation; Above-mentioned eye is meant Eye ointments, ophthalmic emulsion, gel for eye with semi-solid preparation; Above-mentioned eye is meant eye mask agent, eye pill, ophthalmic intercalating agent with solid preparation.
The multi-section position of Chinese medicine, the effect of many target spots have been brought into play in paeonol of the present invention and derivant thereof and peoniflorin combination.The single component of paeonol and derivant thereof and peoniflorin all is useful on the clinical application of non-field of ophthalmology, existing a large amount of report paeonol and peoniflorin safety and low toxicity.Available data [" medical research journal, 2009 38 volume the 4th phase 131-133; " Chinese herbal medicine " 2010 33 the 10th phases of volume, the 1548-1550 page or leaf] show: paeonol is the topmost drug activity composition of Chinese medicine Cortex Moutan, and peoniflorin is the topmost drug activity composition of Radix Paeoniae.From pharmacodynamics test analysis of the present invention, the present invention adopts the main effective ingredient compatibility of Cortex Moutan and Radix Paeoniae to use, and has obvious role in synergism.Existing at present big quantity research proves: the content of paeonol is more obvious than the difference of paeoniflorin content in evident difference, the Cortex Moutan in the Cortex Moutan of the different places of production, different acquisition season and different concocting method gained; Equally, content of paeoniflorin also has notable difference in the Radix Paeoniae of the different place of production, different acquisition season and different concocting method gained.Therefore; The present invention adopts paeonol and peoniflorin to carry out the pharmacological action that the proportioning combination not only is of value to better further investigation paeonol and the different proportionings of peoniflorin; Help the development and utilization of novelty Chinese medicine; And to realizing controllability, the repeatability of Chinese medicine preparation quality, and further investigate its pharmacological action and mechanism of action thereof, development of new Chinese medicine has significant helpfulness.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.
In order to understand essence of the present invention better, pharmacological testing and the result with paeonol and derivant and peoniflorin combination explains its new purposes in pharmaceutical field below.Further paeonol and derivant thereof and peoniflorin are combined in as the application that in the medicine of treating oculopathy same or analogous pharmacology's mechanism is arranged all in protection scope of the present invention.
1, acute toxicity of mice and chronic toxicity test
[" Chinese medicine information " 2009 in the bibliographical information of acute toxicity that has paeonol and paeonol sodium sulfonate now and chronic toxicity test; 26 (1): 20]; Proved that paeonol and paeonol sodium sulfonate whole body use and all not have obvious toxic and side effects, paeonol and sodium paeonol sulfonate have been the human clinical's medication that obtains state approval at present.
The correlational study of peoniflorin shows that also its toxicity is very low, under the normal condition, and no obvious adverse reaction.Mice LD50: intravenous injection is 3530mg/kg, and lumbar injection is 9530mg/kg.Peoniflorin has been approved for the new drug of clinical treatment rheumatoid arthritis as II class Western medicine, be the human clinical's medication that obtains state approval.
2, be exposed to outer anatomical physiology characteristic because the eye conjunctival tissue has rich blood vessel, position, make it receive the stimulation of external antigen tissue the most easily and anaphylaxis takes place.According to bibliographical information; Anaphylaxis conjunctivitis has influenced about 1/4 general population (the eye Claritin compares the effect of people's conjunctiva mastocyte, ophthalmology magazine, 2007 16 the 4th phases of volume); Therefore, allergia (immunity) conjunctivitis is commonly encountered diseases, frequently-occurring disease clinically.Because the eye conjunctiva has abundant lymphocyte, mastocyte etc.; When eye receives the stimulation of external antigen tissue and anaphylaxis takes place; Conjunctiva exists the infiltration of oxyphil cell, neutrophil cell and the activation of mastocyte, and wherein the density of the mastocyte in the conjunctiva can be up to 5000/mm
2And the activation of mastocyte and to take off granule be anaphylaxis conjunctivitis, long-term most important link in the pathogenesis of immunity conjunctivitis such as anaphylaxis conjunctivitis, seasonal allergic conjunctivitis, vernal keratoconjunctivitis, macropapillary conjunctivitis; Histamine is to cause the most important medium of immunity conjunctivitis sings and symptoms.Therefore, histamine and mastocyte are treatment eye important goal hypersensitive.Eye drop (Western medicine) overwhelming majority who is used for clinical multiple treatment immunity conjunctivitis at present is through the activation of blocking-up mastocyte and takes off granule and the antihistamine link is brought into play therapeutical effect, but all there is side effect in various degree in these medicines.In the experimental study of the therapeutical effect of active allergia (the claiming immunity again) conjunctivitis that the medicine that the present invention is mixed with brings out ovalbumin, find; The ophthalmic preparation dissimilar, variable concentrations of paeonol and peoniflorin formulated in combination all can significantly alleviate or eliminate the inflammatory reaction of the eye local organization of immunity conjunctivitis animal model, obviously alleviate or eliminate in the conjunctival epithelial cell and substrate in oxyphil cell's infiltration, obviously reduce mast cell degranulation.Find that in the experimental study of the therapeutical effect of allergia (the claiming immunity again) conjunctivitis that the medicine that the present invention is mixed with causes histamine the ophthalmic preparation dissimilar, variable concentrations of paeonol and peoniflorin formulated in combination all can significantly alleviate or eliminate the inflammatory reaction of the eye local organization of immunity conjunctivitis animal model, obviously reduce the permeability of eyelid and conjunctiva blood vessel.The inventor's result of study explanation paeonol and derivant thereof are embodied in antihistamine with the main pharmacological of peoniflorin combination, stablize mastocyte, minimizing mast cell degranulation, minimizing oxyphil cell's infiltration; Can alleviate or eliminate the inflammatory reaction of the eye local organization of the immunity conjunctivitis animal model that active immunity membranous conjunctivitis that ovalbumin brings out and histamine cause significantly; And its effect is superior to single preparation with paeonol or peoniflorin preparation; The main pharmacological that the combination of paeonol of the present invention and derivant thereof and peoniflorin is described has been directed against the most basic pathogenesis of immunity conjunctivitis exactly; And have antihistamine, stablize the mastocyte dual function; Paeonol and derivant thereof and peoniflorin compatibility are used and are had obvious synergistic function aspect drug action; Therefore not only can the efficacious therapy anaphylaxis conjunctivitis, oculopathy such as immunity conjunctivitis such as anaphylaxis conjunctivitis, seasonal allergic conjunctivitis, vernal keratoconjunctivitis, macropapillary conjunctivitis, and the effect of paeonol and derivant and peoniflorin Combination application throughout the year are than application paeonol or peoniflorin are better separately.
3, the inventor uses document public reported " method of sunkening cord under the conjunctiva " and makes the chronic conjunctivitis model; Be about to new zealand white rabbit and make the chronic conjunctivitis animal model with the legal system of sunkening cord under the conjunctiva; And the inflammation situation of employing slit lamp observation animal eye; Result of study shows the inflammatory reaction that the medicament for the eyes of paeonol (or sodium paeonol sulfonate) and peoniflorin formulated in combination all can significantly alleviate chronic conjunctivitis animal model eye; These effects have been directed against the main clinical manifestation of chronic conjunctivitis exactly; And the paeonol (or sodium paeonol sulfonate) of certain matched proportion density and the effect that peoniflorin is combined in some index aspect also are superior to using separately paeonol or use peoniflorin separately; Explain the medicine that is mixed with of the present invention not only can the efficacious therapy chronic conjunctivitis etc. oculopathy, and synergistic function has also been brought into play in paeonol (or sodium paeonol sulfonate) and peoniflorin combination aspect drug action.
4, use classical endotaxin induction uveitis (endotoxin-induced uveitis; EIU) animal model method; Being about to the SD rat adopts abdominal cavity and two metapedes pad injection endotoxins to make the uveitis model; And the inflammatory reaction degree of anterior chamber of eye tissues such as the iris that adopts the slit lamp observation animal is congested, ooze out in the anterior chamber, hypopyon, contracted pupil; The vitreous body of myopia nethike embrane inner surface, the observation paeonol of the methods such as inflammatory infiltration cell counting in the retinal tissue and peoniflorin compositions are to the influence of EIU uveitis rat; Result of study shows the inflammatory reaction that paeonol and peoniflorin compositions can obviously alleviate EIU uveitis rat anterior ocular segment; Reduce in the vitreous body of near-sighted nethike embrane inner surface and the inflammatory cell infiltration in the retinal tissue, these effects have been directed against uveitic main pathological change exactly, and the drug action of paeonol and peoniflorin combination also is superior to using separately paeonol or peoniflorin; Explain the medicine that is mixed with of the present invention can the efficacious therapy uveitis etc. oculopathy, obvious role in synergism has also been brought into play in the combination of paeonol and peoniflorin aspect drug action.
Can draw from above result; The most outstanding advantage of the present invention is: the present invention has excavated new medical application to the combination of known compound paeonol and paeonol derivative and peoniflorin---and paeonol and paeonol derivative thereof and peoniflorin are combined in preparation and treat the application in the medicine of oculopathy, have opened up a new application; Paeonol of the present invention and derivant thereof and peoniflorin combination are used; Has obvious role in synergism; And help realizing controllability, the repeatability of Chinese medicine preparation quality, and further investigate its pharmacological action and mechanism of action thereof, also help the development and utilization of novelty Chinese medicine; Products material of the present invention source is abundant, inexpensive, and paeonol, sodium paeonol sulfonate and peoniflorin safety and low toxicity, and pharmacological action is strong, is indicating well prospect in medicine.
Below, foregoing of the present invention is further described again through the specific embodiment of embodiment form.But should this be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following instance.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
The specific embodiment
Embodiment 1
The preparation of paeonol and peoniflorin compound eye drops: the method for preparing eye drop according to routine prepares the paeonol and the peoniflorin eye drop of variable concentrations and proportioning ratio respectively.In the preparation process,, also optionally add adjuvant and hyaluronate sodium or viscosity excipient such as methylcellulose or hydroxypropyl methylcellulose such as Borneolum Syntheticum, Herba Menthae except that by being routinely added to cosolvent, osmotic pressure regulator, antioxidant, antiseptic, the PH regulator.Its objective is and improve the concentration of medicine in ocular tissue, increase its bioavailability, heighten the effect of a treatment and reduce untoward reaction etc.
Above-mentioned cosolvent is meant: be selected from least a in cyclodextrin, polypropylene based polymers, polyvinyl alcohol, polyvidon, para-amino benzoic acid, the polysorbate, or their mixture of at least two kinds.Osmotic pressure regulator is meant: be selected from least a in sodium chloride, borate, the boric acid, or their mixture of at least two kinds; Antioxidant is meant: be selected from least a in sodium sulfite, sodium sulfite, sodium pyrosulfite, sodium thiosulfate, sodium citrate, the ascorbic acid, or their mixture of at least two kinds; Antiseptic is meant: be selected from least a in parabens, phenethanol, the benzalkonium bromide, or their mixture of at least two kinds; The PH regulator is meant: be selected from least a in phosphate buffer, borate buffer solution, sodium hydroxide, the sodium bicarbonate, or their mixture of at least two kinds.
Concrete example: the preparation of 0.01% paeonol and 0.01% peoniflorin compound recipe (1: 1) eye drop.In 1000 milliliters of paeonol and peoniflorin compound eye drops, the percentage by weight of each composition that contains is: paeonol 0.01%, peoniflorin 0.01%, polysorbate 0.5%, sodium citrate 0.2%, phenethanol 0.1%, Borneolum Syntheticum 0.5%, sodium chloride is an amount of, sodium hydroxide is an amount of, all the other are sterile purified water.
Sodium hydroxide is meant in right amount: through dripping the 1M sodium hydroxide, make join solution pH value be adjusted into 4.5～7.0.The 1M sodium hydroxide solution prepares in advance, and the 1M sodium hydroxide solution is illustrated in has 1 mole sodium hydroxide in every liter of solution.
Sodium chloride is meant in right amount: be depressed into the tolerant seepage of eyes through the seepage that adds an amount of sodium chloride solution that adjustment is joined and press scope, the seepage that is equivalent to the sodium chloride solution of 0.6%-1.5% like the tolerant seepage pressure of eyes scope is pressed.
Manner of formulation at present embodiment is: after 1) paeonol, peoniflorin, polysorbate, sodium citrate, phenethanol, Borneolum Syntheticum being prepared with corresponding percentage by weight, use the aseptic distillation water dissolution, through stirring it is dissolved fully; 2) with the 1M sodium hydroxide solution its pH value is adjusted into 4.5～7.0; 3) add sterile purified water to 1000 milliliter, add an amount of sodium chloride and regulate seepage and press, stir, and be filtered to clarification, clear filtrate is sub-packed in aseptic plastics eyedrops bottle and promptly gets.Said process carries out under aseptic condition.
The method for preparing of the paeonol of other concentration and different proportionings and peoniflorin compound eye drops similarly, the deal of paeonol and peoniflorin is different and different according to concentration, other components in proportions is constant also can do suitable adjustment according to purpose and needs.
Embodiment 2
The preparation of sodium paeonol sulfonate and peoniflorin compound eye drops: the method preparation for preparing eye drop according to routine.Its preparation method and process are said and get according to embodiment 1.Wherein component paeonol among the embodiment 1 is replaced with sodium paeonol sulfonate, each component ratio of replacement back and content need ratio to measure in sodium paeonol sulfonate and each component of peoniflorin compound eye drops to get final product.
Embodiment 3
The preparation of paeonol and peoniflorin compound recipe eye ointment: the method for preparing eye ointment according to routine prepares the paeonol and the peoniflorin compound recipe eye ointment of variable concentrations and proportioning ratio respectively.In the preparation process, can select different mastic such as lanoline, Yellow Vaselin, liquid paraffin or its mixture etc. for use, also optionally add adjuvants such as Borneolum Syntheticum, Herba Menthae, cyclodextrin.Its objective is and improve the concentration of medicine in ocular tissue, increase its bioavailability, heighten the effect of a treatment and reduce untoward reaction etc.
Concrete example: the preparation of 2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment.In 1000 gram paeonol and peoniflorin compound recipe eye ointment, contain paeonol 22.5 grams, peoniflorin 7.5 grams, Borneolum Syntheticum 5 grams, all the other are the eye pasting substrate of Yellow Vaselin, lanoline, liquid paraffin composition.
Each components in proportions of above-mentioned eye pasting substrate is: Yellow Vaselin: lanoline: liquid paraffin=8: 1: 1.
Manner of formulation at present embodiment is: 1) takes by weighing paeonol 22.5 grams, peoniflorin 7.5 grams, Borneolum Syntheticum 5 grams, grinds, is screened to below the 75 μ m, and subsequent use; 2) with above-mentioned eye pasting substrate heating and melting, filtration, reheat to 130 ℃～150 ℃ sterilization 60～80 minutes is cooled to room temperature, and is subsequent use; 3) paeonol, peoniflorin and the Borneolum Syntheticum after grinding is sieved is moistening with an amount of sterilized liquid paraffin, adds sterilization matrix to 1000 gram, stirs well, fill promptly gets.Whole process for preparation is operated under aseptic condition.
The method for preparing of the paeonol of other concentration and proportioning and peoniflorin compound recipe eye ointment similarly, the deal of paeonol and peoniflorin is different and different according to concentration and proportioning, other components in proportions is constant also can do suitable adjustment according to purpose and needs.
Embodiment 4
The preparation of 1% paeonol eye ointment: the paeonol eye ointment for preparing the method preparation 1% of eye ointment according to routine.That is: in 1000 gram paeonol eye ointment, contain paeonol 10 grams, Borneolum Syntheticum 5 grams, all the other are the eye pasting substrate of Yellow Vaselin, lanoline, liquid paraffin composition.Its preparation method and process are said and get according to embodiment 3.
Embodiment 5
The preparation of 1% peoniflorin eye ointment: the red peoniflorin eye ointment for preparing the method preparation 1% of eye ointment according to routine.That is: in 1000 gram peoniflorin eye ointment, contain peoniflorin 10 grams, Borneolum Syntheticum 5 grams, all the other are the eye pasting substrate of Yellow Vaselin, lanoline, liquid paraffin composition.Its preparation method and process are said and get according to embodiment 3.
Embodiment 6
The preparation of paeonol and peoniflorin compound oral administration preparation: will directly buy commercially available paeonol by the industry standard preparation and peoniflorin (unit: g) product is pressed following parts by weight proportioning:
| Medical material | Prescription 1 | Prescription 2 | Prescription 3 |
| Paeonol | ?50 | ?60 | ?80 |
| Peoniflorin | ?50 | ?30 | ?20 |
After the paeonol of different proportionings, peoniflorin mixed, process various preparations according to the conventional method for preparing of Chinese medicine, like tablet, capsule, granule etc.
Embodiment 7
The research of the therapeutical effect of the active immunity membranous conjunctivitis that the present invention brings out ovalbumin.
1, test material
Tried the drug group medication: 0.25% paeonol and 0.5% peoniflorin (0.5: 1) compound eye drops, 0.01% paeonol and 0.01% peoniflorin (1: 1) compound eye drops, 0.5% paeonol and 0.25% peoniflorin (2: the 1) compound eye drops processed according to embodiment 1 said method; 1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: the 1) compound eye drops processed according to embodiment 2 said methods; 2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment, 2% paeonol and 0.5% peoniflorin (4: 1) the compound recipe eye ointment processed according to embodiment 3 said methods processed according to embodiment 3 said methods.Matched group medication: 2% sodium chromoglicate eye drop, 1% paeonol eye ointment (by embodiment 4 gained), 1% peoniflorin eye ointment (making) by embodiment 5.
2, test method
(1) modeling method
Selecting body weight for use is SD rat, the male and female half and half of 100g～120g.Adopt " the active allergic conjunctivitis that ovalbumin brings out " animal modeling method of open source literature report, the SD rat is made as the rat model of immunity (claiming allergia again) conjunctivitis.
(2) test is divided into groups
55 rats that will be made into immunity conjunctivitis animal model at random are divided into 11 groups at random, every group of 5 animals (10 eyes): model contrasts 1 group (solvent group of sodium paeonol sulfonate and peoniflorin compound eye drops); Model contrasts 2 groups (matrix group of paeonol and peoniflorin compound recipe eye ointment); 1 group of positive control (2% sodium chromoglicate eye drop group); 2 groups of positive controls (1% paeonol eye ointment group); 3 groups of positive controls (1% peoniflorin eye ointment group); Receive 1 group of reagent thing [0.25% paeonol and 0.5% peoniflorin (0.5: 1) compound eye drops]; Receive 2 groups of reagent things [0.01% paeonol and 0.01% peoniflorin (1: 1) compound eye drops]; Receive 3 groups of reagent things [0.5% paeonol and 0.25% peoniflorin (2: 1) compound eye drops]; Receive 4 groups of reagent things [2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment]; Receive 5 groups of reagent things [2% paeonol and 0.5% peoniflorin (4: 1) compound recipe eye ointment]; Receive 6 groups of reagent things [1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: 1) compound eye drops].
(3) medication
1 group of positive control: with 2% sodium chromoglicate eye drop.The local beginning of the 12nd day eye behind rats by intraperitoneal injection ovalbumin phosphate buffer continuous use 3 days, 4 times/day, i.e. 8am, 12am, 4pm, 8pm; The local antigen challenge of eye is carried out in behind rats by intraperitoneal injection ovalbumin phosphate buffer the 15th day; Beginning in 5 minutes eye local application once more after the local antigen challenge of eye; Per 10 minutes once, totally three times (i.e. eye local antigen challenge after the 5th minute, the 15th minute, the 25th minute).
Model contrasts 1 group: with the solvent eye drip of preparation paeonol and peoniflorin compound eye drops, concrete medication number of times is with 1 group of positive control.
Receive 1 group of reagent thing: 0.25% paeonol and 0.5% peoniflorin (1: 2) compound eye drops, concrete medication number of times is with 1 group of positive control.
Receive 2 groups of reagent things: 0.01% paeonol and 0.01% peoniflorin (1: 1) compound eye drops, concrete medication number of times is with 1 group of positive control
Receive 3 groups of reagent things: 0.5% paeonol and 0.25% peoniflorin (2: 1) compound eye drops, concrete medication number of times is with 1 group of positive control.
Receive 6 groups of reagent things: 1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: 1) compound eye drops, concrete medication number of times is with 1 group of positive control.
Model contrasts 2 groups: the substrate with preparation paeonol and peoniflorin compound recipe eye ointment is coated with eye, and the 12nd day eye behind rats by intraperitoneal injection ovalbumin phosphate buffer is local to begin continuous use 3 days, and 2 times/day, i.e. 8am, 8pm; The local antigen challenge of eye is carried out in behind rats by intraperitoneal injection ovalbumin phosphate buffer the 15th day, after the local antigen challenge of eye, begins medication once more 1 time in 5 minutes (promptly after the local antigen challenge the 5th minute).
2 groups of positive controls: with 1% paeonol eye ointment, concrete medication number of times contrasts 2 groups with model.
3 groups of positive controls: with 1% peoniflorin eye ointment, concrete medication number of times contrasts 2 groups with model.
Receive 4 groups of reagent things: 2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment; Concrete medication number of times contrasts 2 groups with model.
Receive 5 groups of reagent things: 2% paeonol and 0.5% peoniflorin (4: 1) compound recipe eye ointment; Concrete medication number of times contrasts 2 groups with model.
(4) index detects
1. the observation of the eye local inflammation extent of reaction: after local antigen challenge, carried out the slit lamp tissue reaction's degree that judges in the 10th minute, the 20th minute, the 30th minute.With following scoring method the extent of reaction is carried out quantitatively.
(palpebral conjunctiva and bulbar conjunctiva) 0 minute: normal; 1 minute: the conjunctiva pinkiness; 2 minutes: conjunctiva took on a red color; 3 minutes: conjunctiva was kermesinus, and ecchymosis, petechia are arranged.
Edema 0 minute: normal; 1 minute: palpebra inferior edema only; 2 minutes: going up palpebra inferior all had edema, and the eyelid part is closed; 3 minutes: ectropion of lid, edema is obvious, and eyelid is semi-closed at least; 4 minutes: go up palpebra inferior and the equal edema of margo palpebrae.
Secretions 0 minute: do not have; 1 minute: rare mucus shape; 2 minutes: eyelid and hair on every side were moist; 3 minutes: eyelid and hair on every side were moist, thick mucus shape
2. histological examination: 35 minutes excision upper ball conjunctivas prepare light microscopy specimen after local antigen challenge, and in Karnovsky ' s fixative, the ethanol gradient is dewatered with fixation of tissue; The embedding of methacrylate ethanol; Slice thick 2 μ m carry out HE, PAS, Giemsa dyeing respectively, om observation.Use grid as the netted counting chi of 0.25mm * 0.25mm in 3 different visuals field respectively to conjunctival epithelium, on acidophil (Eos), mastocyte (MC) sum, degranulated MC counting in subcutaneous tissue and the matrix organization, calculate the counting of meansigma methods then as this BIAO and BEN.
(5) result:
1. model contrast 1 group with model contrast 2 groups after local antigen challenge; Immunity anaphylaxiss such as significantly hyperemia, edema, secretions appear in the anterior ocular segment tissue; And hyperemia, edema, secretions increase the weight of gradually, after local antigen challenge, peak in 30 minutes; Respectively tried drug group and positive control 1-3 group and all the immunity anaphylaxis (hyperemia, edema, secretions) of immunity conjunctivitis rat anterior ocular segment tissue is had the obvious suppression effect in each period; The hyperemia of anterior ocular segment tissue, edema, secretions all are the change that alleviates gradually; Wherein, receive 1 group of reagent thing, this effect that receives 2 groups of reagent things and 1 group of (sodium chromoglicate eye drop) basic identical, its difference not statistically significant (p＞0.05) of positive control; Receive this effect of reagent thing 3-6 group obviously to be superior to 1 group of positive control (sodium chromoglicate eye drop), positive control 2 groups (1% paeonol eye ointment group) and positive control 3 groups of (1% peoniflorin eye ointment group), its differences all have statistical significance (p＜0.05, p＜0.01) (seeing table 1-3).Result of the test shows: the variable concentrations in this test and the paeonol of proportioning and peoniflorin compound eye drops and eye ointment, 1.5% paeonol yellow acid receive and 0.75% peoniflorin eye drop all can be effectively to the immunity anaphylaxis of anti-immune conjunctivitis anterior ocular segment tissue; Proportioning is that 2: 1,3: 1,4: 1 paeonol and peoniflorin compound recipe ophthalmic preparation and proportioning are that 2: 1 the sodium paeonol sulfonate and the curative effect of peoniflorin eye drop are superior to 1% paeonol eye ointment and 1% peoniflorin eye ointment in this test.Explain paeonol and derivant thereof and peoniflorin compatibility use can not only be effectively to the immunity anaphylaxis of anti-immune conjunctivitis anterior ocular segment tissue, and have obvious role in synergism.
2. model contrasts 1 group and contrasts in 2 groups the conjunctival tissue oxyphil cell with model and soak in a large number, degranulated mastocyte showed increased, and the ratio of degranulated mastocyte and mastocyte sum obviously increases; Respectively tried the quantity of degranulated mastocyte and acidophil in the conjunctival tissue that drug group and positive control 1-3 group eye liquid all can obviously reduce immunity conjunctivitis rat; Wherein, receive 1 group of reagent thing, this effect that receives 2 groups of reagent things and 1 group of (sodium chromoglicate eye drop) basic identical, its difference not statistically significant (p＞0.05) of positive control; Receive this effect of reagent thing 3-6 group obviously to be superior to 1 group of positive control (sodium chromoglicate eye drop), positive control 2 groups of (1% paeonol eye ointment group) and 3 groups of positive controls (1% peoniflorin eye ointment group), its difference all has statistical significance (p＜0.05) (seeing table 4).Result of the test shows: the variable concentrations in this test and the paeonol of proportioning and peoniflorin compound recipe eye liquid and ointment, 1.5% paeonol yellow acid are received and 0.75% peoniflorin eye drop suppresses the immunity anaphylaxis of immunity conjunctivitis anterior ocular segment tissue and the granule that takes off that mastocyte is soaked into, reduced to the interior oxyphil cell of its minimizing conjunctival tissue in a large number has obvious relation.Therefore; Paeonol and derivant thereof and peoniflorin compatibility use and all can effectively treat the immunity conjunctivitis in this experiment; And the therapeutical effect of paeonol and peoniflorin combination, sodium paeonol sulfonate and peoniflorin combination is superior to independent paeonol or peoniflorin; Be that paeonol and peoniflorin compatibility, paeonol derivative and peoniflorin compatibility have obvious role in synergism, and be superior to the sodium chromoglicate eye drop at finite concentration and its effect of ratio range.
Table 1 receives the reagent thing that (n=10) compared in the scoring of schematic eye chemosis degree affect
Annotate: (X ± SD, statistical method: adopt SPSS13.0 software, relatively use one factor analysis of variance between the many groups of identical index, between many groups in twos relatively with the q check, below each epiphase with).
Positive in organizing 1 group, receive reagent thing 1-3 group, receiving 6 groups of reagent things and model to contrast 1 group relatively
▲P＜0.05,
▲ ▲P＜0.01; Positive in organizing 2,3 groups, receiving 4,5 groups of reagent things and model to contrast 2 groups relatively
△P＜0.05,
△ △P＜0.01; Respectively being tried drug group and positive control compares for 1 group
P＜0.05,
P＜0.01; Respectively being tried drug group and positive control compares for 2 groups
★P＜0.05,
★ ★P＜0.01; Respectively being tried drug group and positive control compares for 3 groups
◆P＜0.05,
◆ ◆P＜0.01
Table 2 receives the reagent thing that (n=10) compared in the scoring of schematic eye chemosis degree affect
Notes are with table 1.
Table 3 receives the reagent thing that (n=10) compared in the scoring of schematic eye secretions degree affect
Notes are with table 1.
Table 4 receives the counting influence (n=10) of reagent thing to mastocyte (MC) in the schematic eye conjunctival tissue and acidophil (Eos)
Notes are with table 1
Embodiment 8:
The research of the therapeutical effect of the immunity conjunctivitis that the present invention causes histamine.
1, test material
With embodiment 7.
2, test method
(1) modeling method
Selecting body weight for use is SD rat, the male and female half and half of 100g～120g.Adopt " allergic conjunctivitis that histamine causes " animal modeling method of open source literature report, rat is made as the animal model of immunity (claiming allergia again) conjunctivitis.
(2) test is divided into groups
55 rats that will be made into immunity conjunctivitis animal model at random are divided into 11 groups at random, and every group of 5 animals (10 eyes) are specifically tested group technology with embodiment 7.
(3) test procedure
Concrete medication is with embodiment 7.After the last administration 30 minutes, inject 1% diazo blue normal saline solution according to rat body weight to rat tail vein respectively, dosage is the 0.4ml/100g body weight, anesthetized animal after 5 minutes is attacked with histamine 300ng/10 μ g subconjunctival injection immediately; The 5th minute behind the subconjunctival injection histamine, the 15th minute under slit lamp microscope to eye irritated inflammatory reaction observe evaluation, behind the subconjunctival injection histamine, put to death animal in 20 minutes, extract eyelid and conjunctiva.
(4) index detects
1. the observation of the eye local inflammation extent of reaction: with carrying out the slit lamp tissue reaction's degree that judges in the 5th minute, the 15th minute after the histamine subconjunctival injection attack.With scoring method hyperemia, edema and the partial secretions extent of reaction of eye of palpebral conjunctiva and bulbar conjunctiva are carried out quantitatively, concrete point system is with above-mentioned embodiment 7.
2. the quantitative assay of vascular permeability change: attacking back 20 minutes execution animals with the histamine subconjunctival injection, extract eyelid and conjunctiva, remove fatty tissue.To organize according to zoological specimens numberings and to place 10ml tool plug centrifuge tube respectively; (V: V) acetone and mixed liquor of normal saline 4ml soak vibration to add 1: 1; After 24 hours with solution centrifugal 15 minutes in 3000r/min; Draw supernatant and measure its absorbance in the 610nm wavelength, organize a significance relatively with the OD value with spectrophotometer.
(5) result:
1. model contrasts 1 group and contrasts 2 groups after local antigen challenge with model, and immunity anaphylaxiss such as tangible hyperemia, edema, secretions appear in anterior ocular segment tissue, and increase the weight of gradually; Respectively tried drug group and positive control 1-3 group and all the immunity anaphylaxis (hyperemia, edema, secretions) of immunity conjunctivitis rat anterior ocular segment tissue is had the obvious suppression effect in each period; The hyperemia of anterior ocular segment tissue, edema, secretions all are the change that alleviates gradually, receive 1 group of reagent thing, receive this effect and 1 group of (2% sodium chromoglicate eye drop) basic identical, its difference not statistically significant (p＞0.05) of positive control of 2 groups of reagent things; This effect that organized by reagent thing 3-6 obviously is superior to 1 group of positive control (2% sodium chromoglicate eye drop), its difference has statistical significance (p＜0.05, p＜0.01); This effect that organized by reagent thing 3-6 obviously is superior to 2 groups of positive controls (1% paeonol eye ointment group), its difference has statistical significance (p＜0.05); This effect that organized by reagent thing 3-6 obviously is superior to 3 groups of positive controls (1% peoniflorin eye ointment group), its difference has statistical significance (p＜0.05, p＜0.01) (seeing table 5-7).Result of the test shows: the variable concentrations in this test and the paeonol of proportioning and peoniflorin compound recipe eye liquid and eye ointment, the paeonol yellow acid is received and the peoniflorin compound eye drops all can be effectively to the immunity anaphylaxis of anti-immune conjunctivitis anterior ocular segment tissue, proportioning is that 2: 1,3: 1,4: 1 paeonol and peoniflorin compound recipe ophthalmic preparation and proportioning are that 2: 1 the sodium paeonol sulfonate and the curative effect of peoniflorin eye drop are superior to 1% paeonol eye ointment and 1% peoniflorin eye ointment in this test.Explain paeonol and derivant thereof and peoniflorin compatibility use can not only be effectively to the immunity anaphylaxis of anti-immune conjunctivitis anterior ocular segment tissue; And the therapeutical effect of paeonol and peoniflorin combination, sodium paeonol sulfonate and peoniflorin combination is superior to independent paeonol or peoniflorin; Be that paeonol and peoniflorin compatibility, paeonol derivative and peoniflorin compatibility have obvious role in synergism, and be superior to the sodium chromoglicate eye drop at finite concentration and its effect of ratio range.
2. contrast 1 group and model contrast 2 comparisons with model, respectively tried the OD value that drug group and positive control 1-3 group all can obviously reduce the eyelid and the conjunctival tissue of schematic eye.Wherein, receive 1 group of reagent thing and this effect and 1 group of (2% sodium chromoglicate eye drop) basic identical, its difference not statistically significant (p＞0.05) of positive control of receiving 2 groups of reagent things; This effect that organized by reagent thing 3-6 obviously is superior to 1 group of positive control (2% sodium chromoglicate eye drop), its difference has statistical significance (p＜0.05, p＜0.01); This effect that organized by reagent thing 4-6 obviously is superior to 2 groups of positive controls (1% paeonol eye ointment group), its difference has statistical significance (p＜0.05); This effect that organized by reagent thing 3-6 obviously is superior to 3 groups of positive controls (1% peoniflorin eye ointment group), its difference has statistical significance (p＜0.05, p＜0.01) (seeing table 8).Result of the test shows: the variable concentrations in this test and the paeonol of proportioning and peoniflorin compound eye drops and eye ointment, 1.5% paeonol yellow acid are received and 0.75% peoniflorin eye drip fluid power is through reducing the vascular permeability of eyelid and conjunctival tissue to the anaphylactoid main mechanism of immunity of the inductive immunity conjunctivitis of antihistamine local organization effectively.Therefore; Paeonol and derivant thereof and peoniflorin compatibility use and all can effectively treat the immunity conjunctivitis in this experiment; And the therapeutical effect of paeonol and peoniflorin combination, sodium paeonol sulfonate and peoniflorin combination is superior to independent paeonol or peoniflorin; Be that paeonol and peoniflorin compatibility, paeonol derivative and peoniflorin compatibility have obvious role in synergism, and be superior to the sodium chromoglicate eye drop at finite concentration and its effect of ratio range.
Table 5 receives the reagent thing that (n=10) compared in the scoring of schematic eye conjunctival congestion degree affect
Notes are with table 1
Table 6 receives the reagent thing that (n=10) compared in the scoring of schematic eye chemosis degree affect
Notes are with table 1
Table 7 receives the reagent thing that (n=10) compared in the scoring of schematic eye secretions degree affect
Notes are with table 1
Table 8 receives the influence (n=10) of reagent thing to the schematic eye vascular permeability
Notes are with table 1
Embodiment 9 the present invention are to uveitis (endotoxin-induced uveitis, EIU) research of therapeutical effect of endotaxin induction
1, method:
(1) modeling method
Adopt the method for the endotaxin induction uveitis animal model of public reported, be about to induce uveitis (EIU) model bottom two metapedes that 200 μ g Bacillus typhi endotoxins are injected in the SD rat.
(2) test is divided into groups and medication
70 SD rats that are made into the EIU animal model are divided into 7 groups at random:
The paeonol of 1: 1 proportioning and peoniflorin compound recipe are tried drug group and (are called for short 1: 1 paeonol peoniflorin compound recipe group; 10): inject at the Bacillus typhi endotoxin and to begin to give paeonol 2.5mg/kg/ time+peoniflorin in preceding 1 hour and irritate stomach 2.5mg/kg/ time; Whenever once, totally 4 times at a distance from 4 hours.
The paeonol of 2: 1 proportionings and peoniflorin compound recipe are tried drug group and (are called for short 2: 1 paeonol peoniflorin compound recipe groups; 10); Inject at the Bacillus typhi endotoxin and to begin to give paeonol 5mg/kg/ time+peoniflorin in preceding 1 hour and irritate stomach 2.5mg/kg/ time, every at a distance from 4 hours once, totally 4 times;
The paeonol of 4: 1 proportionings and peoniflorin compound recipe are tried drug group and (are called for short 4: 1 paeonol peoniflorin compound recipe groups; 10); Inject at the Bacillus typhi endotoxin and to begin to give paeonol 6mg/kg/ time+peoniflorin in preceding 1 hour and irritate stomach 1.5mg/kg/ time, every at a distance from 4 hours once, totally 4 times;
Model control group (10) is irritated stomach with isometric normal saline, and the filling stomach time is identical with the treatment group with time number average;
1 group of positive control (10) is injected at the Bacillus typhi endotoxin and to be begun to give meloxicam group 0.3125mg/kg/ time in preceding 1 hour and irritate stomach, every at a distance from 4 hours once, totally 4 times;
2 groups of positive controls (paeonol matched group, 10) are injected at the Bacillus typhi endotoxin and to be begun to give paeonol 7.5mg/kg/ time in preceding 1 hour and irritate stomach, every at a distance from 4 hours once, totally 4 times;
3 groups of positive controls (peoniflorin matched group, 10) are injected at the Bacillus typhi endotoxin and to be begun to give peoniflorin 7.5mg/kg/ time in preceding 1 hour and irritate stomach, every at a distance from 4 hours once, totally 4 times.
(3) index detects
Before Bacillus typhi endotoxin injection, in the injection iris hyperemia with the slit lamp microscope observation rat in back 24 hours, anterior chamber, ooze out, the inflammatory reaction of anterior chamber of eye tissues such as hypopyon, contracted pupil; And judge tissue inflammation reaction's degree with reference to the methods of marking scoring of Rick Hoekzema etc., with following scoring method the inflammatory reaction degree is marked quantitatively.
Iris is congested: 0 minute, do not have congested; 1 minute, mild hyperaemia; 2 minutes, moderate was congested; 3 minutes, severe was congested.
Contracted pupil: 0 minute, acorea dwindled, and 1 minute, contracted pupil.
Ooze out in the anterior chamber: 0 minute, nothing was oozed out; 1 minute, ooze out on a small quantity; 2 minutes, ooze out in a large number.
Hypopyon: 0 minute, do not have; 1 minute, have.
The maximum possible value is 7 minutes.
After the inflammatory reaction degree of back 24 hours slit lamp microscope observation anterior chamber of eye tissues of injection finishes; Put to death rat; Extract eyeball, and eyeball was placed the 40ml/L paraformaldehyde solution fixing 72 hours, make sagittate section through pupil optic disc axle; Do routine paraffin wax embedded section, HE dyeing; Om observation, under high power lens, use grid as the netted counting chi of 0.25mm * 0.25mm in 3 different visuals field respectively to inflammatory infiltration cell counting in vitreous chamber myopia nethike embrane inner surface, the retinal tissue, calculate the counting of meansigma methods then as this BIAO and BEN.
The result: the rat anterior chamber of eye inflammatory reaction degree that 1. 1: 1,2: 1,4: 1 paeonol peoniflorin compound recipe groups, 1 group of positive control, 2 groups of positive controls and positive control are 3 groups is all than the obvious reduction of model control group (P＜0.01, P＜0.05); The inflammatory reaction degree of 2: 1 and 4: 1 paeonol peoniflorin compound recipe group rat anterior chamber of eye all obviously reduces (P＜0.05) (seeing table 9) for 3 groups than 1 group of positive control, 2 groups of positive controls, positive control.Experimental studies results prompting: the paeonol peoniflorin compound recipe of 1: 1,2: 1,4: 1 proportionings and meloxicam, paeonol, peoniflorin are congested to the iris of the uveitis SD rat of Bacillus typhi endotaxin induction, ooze out in the anterior chamber, hypopyon, contracted pupil inflammatory reaction all have the obvious suppression effect; And the therapeutical effect of 2: 1,4: 1 paeonol peoniflorin compound recipes is superior to using separately paeonol or uses peoniflorin separately; Explain that paeonol peoniflorin compound recipe has the effect of obvious suppression acute uveitis, paeonol and peoniflorin compatibility use and also have synergistic function simultaneously.2. 1: 1,2: 1,4: 1 paeonol peoniflorin compound recipe and meloxicam, paeonol, peoniflorin all can obviously reduce inflammatory infiltration cell in vitreous chamber myopia nethike embrane inner surface and the retinal tissue of uveitis SD rat of Bacillus typhi endotaxin induction; And the therapeutical effect of 2: 1 paeonol peoniflorin compound recipe also is superior to paeonol matched group and Radix Paeoniae matched group glycosides (P＜0.05) (seeing table 10); Explain that paeonol peoniflorin compound recipe not only has the therapeutical effect of obvious suppression acute uveitis, and paeonol and the peoniflorin compatibility use effect of Synergistic in addition.
Table 9 receives the reagent thing that (n=10) compared in the scoring of endotaxin induction uveitis rat anterior chamber of eye inflammation degree affect
Annotate: (X ± SD, statistical method: adopt SPSS13.0 software, relatively use one factor analysis of variance between the many groups of identical index, relatively check in twos between many groups) with q
Compare with model control group
▲P＜0.05,
▲ ▲P＜0.01; Respectively being tried drug group and positive control compares for 1 group
P＜0.05; Respectively receive reagent
Thing group and positive control compare for 2 groups
★P＜0.05; Respectively being tried drug group and positive control compares for 3 groups
◆P＜0.05
Table 10 receives the therapeutical effect (n=10) of reagent thing to endotaxin induction uveitis rat
Notes are with table 9
Embodiment 11 the present invention are to the test of the therapeutical effect of chronic conjunctivitis
1, test material
Tried the drug group medication: 0.01% paeonol and 0.01% peoniflorin (1: the 1) compound eye drops processed according to embodiment 1 said method; 1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: the 1) compound eye drops processed according to embodiment 2 said methods; 2.25% paeonol and 0.75% peoniflorin (3: 1) the compound recipe eye ointment processed according to embodiment 3 said methods.The positive controls medication is respectively: ZHENZHU MINGMU DIYANYE (commercially available), 1% paeonol eye ointment (by embodiment 4 gained), 1% peoniflorin eye ointment (making by embodiment 5).
2, test method
(1) modeling method
New zealand white rabbit, body weight 2.0 ± 0.1kg, male and female half and half, eye does not have illness.The chronic conjunctivitis model is made according to " method of sunkening cord under the conjunctiva " of document public reported.
(2) test is divided into groups: 56 new zealand white rabbits that will be made into the chronic conjunctivitis animal model are divided into 8 groups at random, 7 every group (14 eyes).Model contrasts 1 group (solvent group of paeonol and peoniflorin compound eye drops); Model contrasts 2 groups (matrix group of paeonol and peoniflorin compound recipe eye ointment); 1 group of positive control (ZHENZHU MINGMU DIYANYE group); 2 groups of positive controls (1% paeonol eye ointment group); 3 groups of positive controls (1% peoniflorin eye ointment group); Receive 1 group of reagent thing [0.01% paeonol and 0.01% peoniflorin (1: 1) compound eye drops]; Receive 2 groups of reagent things [1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: 1) compound eye drops]; Receive 3 groups of reagent things [2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment].Begin to drip in the conjunctival sac medicine treatment, 3 times/day, totally 14 days after 24 hours in modeling.
(3) observation of curative effect
1. the 7th day, the 14th day employing slit lamp microscope, pocket lamp bore hole are observed the eye inflammation situation after medication, and with following scoring method the eye inflammation degree are carried out quantitatively.
Margo palpebrae 0 minute is normal; 1 minute, wet, pieces shape; 2 minutes, congested, edema.
Secretions 0 minute does not have; 1 minute, corner of the eyes portion was a small amount of; 2 minutes, conjunctival sac secretions; 3 minutes, margo palpebrae was sticked together.
Congested 0 minute of palpebral conjunctiva does not have; 1 minute, slight; 2 minutes, moderate; 3 minutes, severe.
Congested 0 minute of bulbar conjunctiva does not have; 1 minute, slight; 2 minutes, moderate; 3 minutes, severe.
Chemosis 0 minute does not have; 1 minute, transparent slight protuberance; 2 minutes, bulge; 3 minutes, the blister projection.
2. curative effect judging standard: cure, total points is a 0-0.5 branch produce effects; Total points is 0.5～1.5 minute; Effectively, total points is＜1.5～2.5 minutes; Invalid, total points is more than 2.5 minutes.
3, result
After treatment 7 days, positive control 1-3 group, each trial drug group and model contrast 1 group, model and contrast 2 groups of comparing difference not statistically significants (P＞0.05); Continuously treatment is after 14 days, contrasts 2 groups except that model contrasts 1 group and model, and positive control 1-3 group and the animal pattern eye inflammation that is respectively tried drug group change all has improvement in various degree.Curative effect grows from weak to strong and is followed successively by 1 group of positive control, 3 groups of positive controls, receives 1 group of reagent thing, 2 groups of positive controls, receive 3 groups of reagent things, receive 2 groups of reagent things (seeing table 11).Therefore, the paeonol yellow acid is received and all can effectively be treated chronic conjunctivitis with peoniflorin compound eye drops, paeonol and peoniflorin compound eye drops, paeonol and peoniflorin compound recipe eye ointment; And the curative effect effect of 1.5% sodium paeonol sulfonate and 0.75% peoniflorin (2: 1) compound eye drops and 2.25% paeonol and 0.75% peoniflorin (3: 1) compound recipe eye ointment not only is superior to ZHENZHU MINGMU DIYANYE; And the therapeutical effect that paeonol (or sodium paeonol sulfonate) and peoniflorin compatibility use is superior to independent paeonol or peoniflorin, i.e. paeonol (or sodium paeonol sulfonate) and peoniflorin compatibility use has synergistic function.
Table 11 receives the therapeutical effect (n=14 eye) of reagent thing to the chronic conjunctivitis animal model
Statistical method: adopt rank test
Claims (10)
1. the new medical use of paeonol and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof and peoniflorin are combined in preparation treatment or prevent the application in the medicine of oculopathy.
2. the new medical use of paeonol according to claim 1 and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof and peoniflorin are combined in preparation and treat the application in the medicine of immunity conjunctivitis.
3. the new medical use of paeonol according to claim 2 and derivant thereof and peoniflorin combination; It is characterized in that: described immunity conjunctivitis is called allergic conjunctivitis again, mainly comprises anaphylaxis conjunctivitis, long-term anaphylaxis conjunctivitis, seasonal allergic conjunctivitis, vernal keratoconjunctivitis, macropapillary conjunctivitis, eczematous keratoconjunctivitis, autoimmune membranous conjunctivitis.
4. the new medical use of paeonol according to claim 1 and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof and peoniflorin are combined in preparation and treat the application in the medicine of chronic conjunctivitis.
5. the new medical use of paeonol according to claim 1 and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof and peoniflorin are combined in the application in the uveitic medicine of preparation treatment.
6. according to the new medical use of each described paeonol of claim claim 1 to 5 and derivant thereof and peoniflorin combination, it is characterized in that: said combination comprises: the series derivates of paeonol, paeonol analog structure or paeonol and derivant thereof are no less than series derivates or peoniflorin and the combination that derivant is no less than two kinds mixture thereof of two kinds mixture and peoniflorin, peoniflorin analog structure.
7. the new medical use of paeonol according to claim 6 and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof and peoniflorin make up by weight that the ratio of mark is 0.5: 1 to 4: 1.
8. the new medical use of paeonol according to claim 7 and derivant thereof and peoniflorin combination; It is characterized in that: the combination of said paeonol and derivant thereof and peoniflorin can be prepared from pharmaceutical composition or the preparation that paeonol and derivant thereof and peoniflorin make up with pharmaceutically acceptable one or more carriers or adjuvant or complementary composition, and the weight percentage that paeonol and derivant thereof and peoniflorin are combined in compositions or the preparation is 0.02% to 99%.
9. the new medical use of paeonol according to claim 8 and derivant thereof and peoniflorin combination is characterized in that: paeonol and derivant thereof with peoniflorin combination can be prepared into eye with liquid preparation, eye with semi-solid preparation, eye with solid preparation, with sustained release pharmaceutical formulation, with film controlled release medicine preparation, sclera release preparation, capsule, pill, tablet, drop pill, granule, injection or oral liquid are used to treat the medicine of oculopathy thoroughly.
10. the new medical use of paeonol according to claim 9 and derivant thereof and peoniflorin combination is characterized in that: said paeonol and derivant thereof become weight percentage >=0.02% of eye with paeonol in the liquid preparation and derivant and peoniflorin combination with the peoniflorin combined preparation.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105796977A (en) * | 2016-04-29 | 2016-07-27 | 毕宏生 | Pharmaceutical composition for treating uveitis |
| CN108159066A (en) * | 2018-02-02 | 2018-06-15 | 昆明理工大学 | The new application of Paeoniflorin |
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| US20080194700A1 (en) * | 2005-05-06 | 2008-08-14 | Angiolab, Inc. | Use of Paeonol For Inhibiting Angiogenesis or For Enhancing Radiosensitization |
| CN101267779A (en) * | 2005-07-27 | 2008-09-17 | 福罗里达大学研究基金会有限公司 | Use of heat shock to treat ocular disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20080194700A1 (en) * | 2005-05-06 | 2008-08-14 | Angiolab, Inc. | Use of Paeonol For Inhibiting Angiogenesis or For Enhancing Radiosensitization |
| CN101267779A (en) * | 2005-07-27 | 2008-09-17 | 福罗里达大学研究基金会有限公司 | Use of heat shock to treat ocular disease |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105796977A (en) * | 2016-04-29 | 2016-07-27 | 毕宏生 | Pharmaceutical composition for treating uveitis |
| CN108159066A (en) * | 2018-02-02 | 2018-06-15 | 昆明理工大学 | The new application of Paeoniflorin |
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