CN101198346A - LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized - Google Patents
LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized Download PDFInfo
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- CN101198346A CN101198346A CNA2006800216616A CN200680021661A CN101198346A CN 101198346 A CN101198346 A CN 101198346A CN A2006800216616 A CNA2006800216616 A CN A2006800216616A CN 200680021661 A CN200680021661 A CN 200680021661A CN 101198346 A CN101198346 A CN 101198346A
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Abstract
The invention relates to a compound, comprising a LKKTET peptide or excitation agent, an ophthalmic carrier, an anti-microorganism corrosion prevention agent, a hyperosmotic agent for providing the ophthalmic permeation pressure, a comfortable-reinforcing agent, at least one of pH adjusting agent for adjusting the pH of the compound to the desired pH level, and a cushion agent for basically remaining the desired pH level, wherein, the peptide agent comprises amino acid sequence LKKTET or LKKTNT or a variant thereof; and the excitation agent excites the generation of LKKTET, LKKTET peptide or the conservative variant thereof.
Description
Background of invention
The cross reference of related application
The present invention requires the rights and interests of U.S. Provisional Application of submitting on June 17th, 2,005 60/691,261 and the U.S. Provisional Application of submitting on February 28th, 2,006 60/776,947.
Technical field
The present invention relates to the field of LKKTET and/or LKKTNT compositions and method.
Background technology
The albumen that originally extrasin beta 4 is considered to be in external endothelial cell migration and is raised between the idiophase.Extrasin beta 4 separates from thymus at first, is one and has 43 aminoacid, 4.9KDa, ubiquitous polypeptide in multiple tissue.Belong to these proteic several functions and be included in function in endothelial cell differentiation and migration, T cell differentiation, actin sequestration, vascularization and the wound healing.
Identified many T β 4 isomers, itself and known T β 4 aminoacid sequences have about 70% or about 75% or about 80% or higher homology.These isomers comprise that for example T β 4
Ala, T β 9, T β 10, T β 11, T β 12, T β 13, T β 14 and T β 15.Similar to T β 4, T β 10 and T β 15 isomers also show the activity that can isolate actin.T β 4, T β 10 and T β 15, and these other isomers have an amino acid sequence LKKTET or LKKTNT, as if it participate in the isolation or the combination of modulate actin.Although do not wish to be limited by any specific theory, the activity of the peptide reagent of Miao Shuing is at least in part owing to the anti-inflammatory activity of these reagent herein.β-thymosin has also been regulated actin polymerization (as if for example, the β thymosin passes through to isolate free G-actin and depolymerization F-actin).The ability of T β 4 modulate actin polymerization is attributable to its ability by combination of LKKTET sequence or isolation actin.
This area needs compositions and the method for LKKTET and/or LKKTNT.
Summary of the invention
Comprise peptide reagent or stimulate reagent and eye with the antiseptic of carrier, anti-microbial effect, the high osmotic agent of described compositions eye with osmotic pressure, comfort enhancers are provided, can regulate extremely required of compositions pH with at least a in the acidity of pH level or the alkaline pH regulator with keep the compositions of the buffer agent of required pH level substantially, wherein said peptide reagent comprises amino acid sequence LKKTET or LKKTNT or its variant, and described stimulation reagent stimulates the generation of LKKTET or LKKTNT peptide or its conservative variant.
Detailed Description Of The Invention
For multiple purpose contains the component that one or more may cause the experimenter to damage to many compositionss that the experimenter uses.
For example, antiseptic, as be used in many eyes with and the benzalkonium chloride (BAK) of cosmetic product, when using this product, can cause surface stimulation and/or injury.Syndrome comprises rubescent and uncomfortable.
The eye drop that life-time service contains quaternary ammonium salt (as BAK) can produce significantly and serious gradually ocular surface stimulation and apoptosis.The patient often develops into such as the such symptom of rubescent and common ophthalmic uncomfortable.Peptide reagent of the present invention, extrasin beta 4 (T β 4) for example when contacting ocular tissue with quaternary ammonium salt (as BAK) experimenter is used, can be used for treating or stops the injury of ocular tissue or impaired.
The example that can be used for quaternary ammonium salt of the present invention comprises BAK, cetrimonium bromide (cetrimide), benzoxonium chloride (benzoxonium) and analog thereof.
According to the specific embodiment, this peptide reagent is used with ammonium salt such as the BAK as a same preparation part, for example joins in the preparation that contains BAK or other quaternary ammonium salts as additive.Quaternary ammonium salt such as BAK are at many eye solution, as being commonly used for antiseptic in the glaucoma eye drop.
In another embodiment, this peptide reagent can be before using quaternary ammonium salt such as BAK, and/or use during quaternary ammonium salt such as the BAK, and/or use after using quaternary ammonium salt such as BAK.
According to the specific embodiment, the present invention especially can be used for stoping the injury to the ocular tissue of cornea epithelium, and it comprises the apoptosis that stops in this class ocular tissue.For example, the present invention is also as the adjuvant that resists blue or green light eye drop or other eye drops.
One aspect of the present invention provides ophthalmic composition, and it comprises peptide reagent or its conservative variant that contains amino acid sequence LKKTET or LKKTNT.
The present invention provides the method for treatment ocular tissue on the other hand, and it comprises the local application ophthalmic composition to described ocular tissue, and said composition comprises peptide reagent or its conservative variant of amino acid sequence LKKTET or LKKTNT
In the specific embodiment, the invention provides the Therapeutic Method that contacts with the compositions that contains peptide reagent described herein of effective dose by with ocular tissue.The embodiment that directly uses comprises, for example directly uses the solution, lotion, ointment, gel, emulsifiable paste, paste, spray, suspension, dispersion liquid, hydrogel, ointment, oil or the foam that contain peptide formulations described herein and contacts with tissue.
Do not find to be suitable for any particular theory, actin-isolation peptide, can promote ocular tissue's health as extrasin beta 4 (T β 4 or TB4) and other reagent that comprises actin-isolation peptide or fragments of peptides, wherein actin-isolation peptide or fragments of peptides contain amino acid sequence LKKTET or LKKTNT or its conservative variant.
Originally extrasin beta 4 is considered to the albumen that raised during endothelial cell migration and the vitro differentiation.Extrasin beta 4 separates from thymus at first, is one and has 43 aminoacid, 4.9Kda, ubiquitous polypeptide in multiple tissue.Belong to these proteic several functions and be included in function in endothelial cell differentiation and migration, T cell differentiation, actin sequestration, vascularization and the wound healing.
According to the specific embodiment, the present invention preferably is applicable to extrasin beta 4 and/or T β 4 isomers, analog or derivant, comprising N-end variant and the C-end variant of the T β 4 of KLKKTET, LKKTETQ, oxidation, T β 4 sulfoxides, T β 4.
According to the specific embodiment, can be used for compositions of the present invention and comprise peptide reagent, for example extrasin beta 4 and/or T β 4 isomers, analog or derivant are comprising the N-end variant of the oxidised form (comprising T β 4 sulfoxides) of T β 4, T β 4 and C-end variant and the polypeptide or the fragments of peptides that comprise or be made up of amino acid sequence LKKTET and conservative variant thereof basically.International Application PCT/US99/17282 discloses the isomer that can be used for T β 4 of the present invention, and can be used for amino acid sequence LKKTET of the present invention or LKKTNT and its conservative variant, and this application mode is by reference included this paper in.International Application PCT/GB99/00833 (WO99/49883) discloses the extrasin beta 4 that can be used for oxidation of the present invention, and this application mode is by reference included this paper in.Although the present invention is primarily aimed at T β 4 hereinafter and T β 4 isomers are described, but should be understood to peptide that following description is equally applicable to amino acid sequence LKKTET or LKKTNT, comprises or is made up of LKKTET, LKKTNT basically and fragment, their conservative variant, and/or T β 4 isomers, analog or derivant, comprising N-end variant and the C-end variant of T β 4, the T β 4 of oxidation.
According to the specific embodiment, compositions of the present invention is used with the frequency in every day, per two days, per two weeks, per two months etc., wherein in the every day of using, can carry out the single or multiple medication, for example whenever uses medication on the same day 2,3,4 or more times.
Identified many T β 4 isomers, itself and known T β 4 aminoacid sequences have about 70% or about 75% or about 80% or higher homology.These isomers comprise, for example T β
4ala, T β 9, T β 10, T β 11, T β 12, T β 13, T β 14 and T β 15.Similar to T β 4, isomer T β 10 and T β 15 also show the activity that can isolate actin.As if T β 4, T β 10 and T β 15, and other isomer consensus amino acid sequences LKKTET or LKKTNT, it participates in the isolation or the combination of modulate actin.T β 4 has anti-inflammatory activity, and also can modulate actin polymerization (as if for example, the β thymosin passes through to isolate free G-actin and depolymerization F-actin).The ability of T β 4 modulate actin polymerization is attributable to its ability by LKKTET or combination of LKKTNT sequence or isolation actin.Like this, the same with T β 4, other anti-inflammatory albumen and/or in conjunction with or isolate actin, perhaps the albumen of modulate actin polymerization (comprising T β 4 isomers with amino acid sequence LKKTET) is independent or make up with T β 4 and play a role probably as this paper explanation.
Therefore, can clearly expect known T β 4 isomers, for example T β 4
Ala, T β 9, T β 10, T β 11, T β 12, T β 13, T β 14 and T β 15, and also unidentified T β 4 isomers can be used for method of the present invention.Like this, T β 4 isomers can be used for method of the present invention, comprising the method that adopts in the experimenter.Thereby the present invention further provides and has comprised T β 4, and T β 4 isomer T β 4
Ala, T β 9, T β 10, T β 11, T β 12, T β 13, T β 14 and T β 15, and eye is with the pharmaceutical composition of carrier.
In addition, as as shown in suitable isolation, combination, mobilization or the polymerization analysis or as by having the bonded aminoacid sequence of modulate actin (for example LKKTETR, LKKTNT) confirms, other have anti-inflammatory activity and/or actin sequestration or binding ability, maybe can mobilize the reagent or the protein of actin or modulate actin polymerization, can similarly be used for method of the present invention.Such albumen comprises for example gelsolin, vitamin D binding protein (DBP), Profilin (profilin), cofilin, depactin (depactin), DNA enzyme I, villin (villin), truncated protein (fragmin), severin (severin), capping protein (capping protein), beta-actin and Acumentin (acumentin).Therefore, method comprises those methods that are used for the experimenter.The present invention further provides pharmaceutical composition, it comprises gelsolin mentioned in this article, vitamin D binding protein (DBP), Profilin (profilin), cofilin, depactin (depactin), DNA enzyme I, villin (villin), truncated protein (fragmin), severin (severin), capping protein (capping protein), beta-actin and Acumentin (acumentin).Therefore, the present invention includes the application of the polypeptide that contains amino acid sequence LKKTET or LKKTNT and its conservative variant.
Term as used herein " conservative variant " or its phraseological variant are meant that amino acid residue is by the biologically similar residue replacement of another kind.The example of conservative variant comprises that for example isoleucine, valine, leucine or methionine are replaced another hydrophobic residue to hydrophobic residue, polar residues is replaced another polar residues, for example replaces lysine, glutamic acid replacement aspartic acid or glutamine with arginine and replaces asparagine and similarly replacement.
According to the specific embodiment, compositions of the present invention is an eye drop preparation.
According to the specific embodiment, be used for compositions of the present invention and comprise peptide reagent described herein, wherein the concentration range of peptide reagent is every milliliter of about 0.001-1000 milligram (mcg/ml), 0.1-100mcg/ml more preferably, most preferably about 1-10mcg/ml.In the particularly preferred specific embodiment, this peptide reagent is T β 4.
According to the specific embodiment, the present invention is used for the treatment of or prevents ocular tissue owing to contact infringement or the damage that causes with quaternary ammonium salt such as BAK, the quaternary ammonium salt concentration range is about 0.0001-1wt%, preferred concentration range is about 0.001-0.1wt%, and preferred concentration range is 0.002-0.05wt%.In a specific embodiment, the concentration of quaternary ammonium salt is about 0.005-0.02wt%.
According to the specific embodiment, the present invention also comprises the pharmaceutical composition that contains peptide reagent described herein, and this peptide reagent can used in the carrier by eye.Such carrier comprises, for example listed herein carrier.
According to the specific embodiment, provide the actual dose of treatment or reagent, preparation or compositions to rely on many factors, comprise experimenter's size and health status.Yet those of ordinary skills can use being used for described in PCT/US99/17282 (seing before) and the list of references quoted to determine that the method and the technology of clinical dosage determine suitable using dosage herein.
According to the specific embodiment, herein the peptide reagent that method and composition used or comprised of Miao Shuing can by with can the eye non-toxic excipients of usefulness or carrier mix and make compositions.
According to the specific embodiment, comprise that the external preparation of reactive compound also comprises the physiological compatibility carrier, the technical staff of field of ophthalmology can use conventional criteria to select this carrier.This carrier is optional uses carrier from known eye, and it is including, but not limited to saline solution; Polyether aqueous solution (waterpolyethers) (for example Polyethylene Glycol); Polyethylene (for example polyvinyl alcohol and polyvidon (povidone)); Cellulose derivative, for example methylcellulose and hydroxypropyl emthylcellulose; Petroleum derivative, for example mineral oil and white vaseline; Animal fat is lanoline for example; Acrylate copolymer is carbomer glue (carboxypolymethylene gel) for example; Plant fat, for example Oleum Arachidis hypogaeae semen; And polysaccharide, for example glucosan; And mucopolysaccharide (glycosaminogiycans), for example hyaluronate sodium; And salt, for example sodium chloride and potassium chloride.
According to the specific embodiment, ophthalmic composition helps the part and is used for eyes, especially with solution, suspension, ointment, gel or foamy form.
According to the specific embodiment, the conventional pharmaceutical that is used for corresponding ophthalmic composition can accept excipient and additive is well known by persons skilled in the art, for example those that hereinafter mention, especially carrier, stabilizing agent, solubilizing agent, penetration enhancer, buffer substance, antiseptic, thickening agent, complexing agent and other excipient.The visible United States Patent (USP) 5,134,124 and 4,906,613 of the such additive and the example of excipient.These compositionss are with known method preparation separately, such as forming corresponding ophthalmic composition by active component is mixed with corresponding excipient and/or additive.Active component is preferably used with the form of eye drop, and active component for example is dissolved in the carrier routinely.Required pH value can suitably be regulated and/or be buffered to this solution, and, add stabilizing agent, solubilizing agent or penetration enhancer in the time of suitably.In the time of suitably, in ophthalmic composition, add antiseptic and/or other excipient.
Employed carrier is applicable to external or common administration usually among the present invention, and for example is water, water and the mixed liquor easy and solvent that water is miscible, for example C
1-C
7-alkanol, the vegetable oil or the mineral oil that contain the 0.5-5wt% hydroxyethyl-cellulose, ethyl oleate, carboxymethyl cellulose, the non-toxic water soluble polymer of polyvinylpyrrolidone and other usefulness, for example cellulose derivative such as methylcellulose, the alkali metal salt of carboxymethyl cellulose, hydroxy methocel, hydroxyethyl-cellulose, methylhydroxypropylcellulose and hydroxypropyl cellulose, acrylate/salt or methacrylate/salt, as polyacrylic salt or ethyl acrylate, polyacrylamide, natural materials such as gelatin, alginate, pectin, tragacanth, karaya, xanthan gum, carrageenin, agar and Radix Acaciae senegalis, starch derivatives such as starch acetate and hydroxypropyl starch, other synthetic products such as polyvinyl alcohol, polyvinylpyrrolidone, polyvinyl methyl ether, polyethylene glycol oxide, preferred cross linked polyacrylate such as neutral carbomer, perhaps these mixture of polymers.Preferred carrier is alkali metal salt, hydroxy methocel, hydroxyethyl-cellulose, methylhydroxypropylcellulose and the hydroxypropyl cellulose of water, cellulose derivative such as methylcellulose, carboxymethyl cellulose, neutral carbomer or its mixture.
According to the specific embodiment, the solubilizing agent that is used for ophthalmic composition of the present invention is that for example tyloxapol, fatty acid glycerine gather the mixture of lower alkyl diol ester (fatty acid glycerol poly-loweralkylene glycol esters), fatty acid polyglycol lower alkyl diol ester (fatty acid poly-loweralkylene glycol esters), Polyethylene Glycol (polyethylene glycols), glycerin ether (glycerolethers) or these chemical compounds.The amount of being added is enough to the lytic activity component usually.For example, the concentration of solubilizing agent be active component concentration 0.1-5000 doubly.Alkylidene in the lower alkyl glycol is meant the alkylidene that comprises 7 carbon atoms at most, for example is methylene, ethylidene, trimethylene, propylene, pentamethylene, 2,5-hexylidene or 1, the inferior heptyl of 7-.Low-grade alkylidene preferably comprises the linearity or the branched alkylidene of 4 carbon atoms at most.
The example of buffer substance is acetate, Ascorbate, borate, bicarbonate/carbonate, citrate, gluconate, Lactobionate, phosphate, propionate and TRIS (trometamol) buffer agent.Trometamol and borate buffer are preferred reducing agents.Add the amount of buffer substance, for example be to guarantee and keep the needed amount of physiological tolerance pH scope.This pH scope is generally 5-9, is preferably 6-8.2, more preferably 6.8-8.1.
Penetration enhancer (tonicity enhancing agent) for example is an ionic compound, as the halogenide of alkali metal or alkaline-earth metal, for example is CaCl
2, KBr, KCl, LiCl, NaBr, NaCl or boric acid.The nonionic penetration enhancer for example is carbamide, glycerol, sorbitol, mannitol, propylene glycol or glucose dextrose.For example add enough penetration enhancers and make the i.e. ophthalmic composition of usefulness, its osmotic pressure approximately is 50-1000mOsmol, 100-400mOsmol preferably, 200-400mOsmol more preferably, 280-350mOsmol more preferably again.
The example of antiseptic is a quaternary ammonium salt, for example the alkyl mercuric salt of cetrimonium bromide, benzalkonium chloride, benzoxonium chloride, thiosalicylic acid, for example thimerosal, phenylmercuric nitrate, phenylmercuric acetate or phenylmercuric borate; P-hydroxybenzoic acid esters, for example nipagin or propyl parabene; Alcohols such as methaform, benzyl alcohol or phenethanol; Guanidine derivatives is as chlorhexidine or poly hexamethylene biguanide, perhaps sorbic acid.Preferred antiseptic is cetrimonium bromide, benzalkonium chloride, benzoxonium chloride and p-hydroxybenzoic acid esters.Suitably in ophthalmic composition, add of the secondary pollution of capacity antiseptic to cause by antibacterial and fungus in guaranteeing to prevent to use.
According to the specific embodiment, this ophthalmic composition can further comprise non-toxic excipients, for example emulsifying agent, wetting agent or filler, and such as Macrogol 200,300,400 and 600, perhaps Carbowax1000,1500,4000,6000 and 10000.If desired, other operable excipient are listed as follows, but they are used to limit the scope of available excipient never in any form.They are chelating agent especially, as EDTA disodium or EDTA; Antioxidant is as ascorbic acid, acetylcysteine, cysteine, sodium sulfite, Butylated hydroxyanisole, butylated hydroxytoluene or alpha-tocopherol acetate; Stabilizing agent is as cyclodextrin, thiourea, sulfo-sorbitol, dioctyl sodium sulfosuccinate or single thioglycerol; Or other excipient, as lauric acid sorbitol ester, Emulphor FM or cetylate.Preferred excipient is a chelating agent, and for example EDTA disodium, and stabilizing agent is as cyclodextrin.Add the amount of excipient and type will be according to specific demand and usually in about scope of 0.0001 to about 90wt%.Cyclodextrin contains a plurality of glucose units, and wherein each glucose all contains 3 free hydroxyl groups.The amount of the cyclodextrin that uses in the specific embodiment is 0.01-20wt% preferably, is more preferably 0.1-15wt%, is more preferably 1-10wt% again.
According to the specific embodiment, the present invention also relates to ophthalmic composition, it comprises the effective pharmaceutical agents of peptide reagent described herein, carrier, solubilizing agent and other treatment for the treatment of effective dose, these pharmaceutical agents can be, for example antibiotic, antiallergic agent, anesthetis, other antiinflammatory, corticosteroid, be applicable to the reagent or the other drug that reduce intraocular pressure (IOP).
PH (hydrogen ion concentration)
According to the specific embodiment, the pH of preparation of the present invention should be as far as possible near the pH of tear film.The physiological pH of tear approximately is 7.4 ± 0.2.Like this, for the consideration to comfortable, tolerance and safety, this is the pH of best ophthalmic preparation.
Stimulate the tear secretion and can cause nictation pH value to reduce.When eyelid opened long-time, the tear film can with the CO in the surrounding air
2Dividing potential drop balances each other and is alkalized, and pH value can be greater than 9.PH reduces and rising does not have adverse consequences.Therefore, when the preparation of statement invention during at about pH7.4, this pH has certain mobility scale.
And when to the eyes administered formulation, it stimulates the outflow tear.Shedding tears can rapid dilution and a spot of substance of buffering, illustrates that eyes can tolerate the wide relatively pH scope that particular formulations causes.
Therefore, the pH value of ophthalmic preparation can be in the scope of about 3.5-11.5.Yet ophthalmic preparation can demonstrate the pH scope of narrower 3.5-9, preferably 4.5-8, most preferably 55.-7.8.Most preferred pH scope has benefited from expection dissolubility, chemical stability and the therapeutic activity of the present composition, and this scope is to stoping useful, the narrow relatively scope of corneal injury.
Buffer system
According to the specific embodiment, buffer system is made up of weak acid or weak base and conjugation salt thereof.The buffer capacity that becomes branch to have in the system, the feasible external action of no matter adding acid or alkali and being subjected to temperature, pressure, volume, oxidation-reduction potential, body fluid and tear, pH will keep substantially constant.Although enough variations (being the pH drift) to resist product pH in rational storage life (promptly under condition of storage) greatly of buffer capacity, even but low also being enough to of the buffer capacity of ophthalmic preparation of the present invention can when ocular administration, can adjust product apace again to physiological pH.According to the specific embodiment, eye should be in the 0.05-1.0 scope with the buffer capacity of product, to specific invention compositions preferably with most preferred buffer capacity respectively in the scope of 0.02-0.2 and 0.01-0.1.Buffer capacity is determined by following formula:
*β＝ΔB/ΔpH
Wherein β is a buffer capacity, Δ B be the strong acid/highly basic pH that changes 1 liter of buffer solution suitable gram number, Δ pH adds the pH variable quantity that strong acid/highly basic caused.
According to the specific embodiment, suitable buffer system can be the sodium salt of following acid: acetic acid, ascorbic acid, boric acid, carbonic acid, phosphoric acid, citric acid, gluconic acid, lactic acid and propanoic acid.The calcium salt of carbonic acid or propanoic acid and the potassium salt of phosphoric acid all can form suitable buffer system.Tris buffer agent (trometamol) uses in vein as the basifier of correcting the metabolism acidosis, and it also is to be used for one of preferred reducing of the present invention.Other preferred reducing agents are acetate, phosphate, citrate and borate.In particular instance, the buffer system that comprises the proton donor of proteinic amino acid residue and proton acceptor is than Acid-Base or ammonia-the alkali buffer agent more preferably.
The concrete consumption of buffer substance can change and depend on to keep the pH environment that is suitable for present composition stability and guarantee and keep physiology and can tolerate the necessary amount of pH scope.
High osmotic agent (tonicity agent)
Listed herein high osmotic agent its objective is at natural tear and regulates the osmotic pressure of ophthalmic composition of the present invention approximately to physiological osmotic pressure (for example 0.9% saline).For example, can in peptide reagent preparation of the present invention, add sodium chloride, potassium chloride, calcium chloride, glucose dextrose and/or mannitol.The amount of high osmotic agent can change, and depends on the concrete reagent that will add.Substantially, the amount of the high osmotic agent that specific invention compositions contains be enough to make final compositions have the eye use acceptable osmotic pressure, osmotic pressure is 150-450mOsm preferably, most preferably 250-350mOsm.
Preferred high osmotic agent is sodium salt and potassium salt, especially sodium chloride and potassium chloride.Most preferred high osmotic agent is a sodium chloride.
Lubricant/wetting agent/viscosity intensifier
According to the specific embodiment, the present invention includes following chemical compound, wherein this chemical compound can make eyes lubricated, reduces surface tension and raising its wettability (contact) of other hydrophobic epithelium anterior corneal surfaces, has roughly concordance with tear.Such chemical compound also can strengthen the viscosity of the present composition, thereby makes the longer peptide reagent that makes of preparation of the present invention time of staying in eyes have more time to bring into play its therapeutic activity or be absorbed and reach required target spot.
Suitable viscosity intensifier and its concentration range in some compositions of the present invention be including, but not limited to (a) monomer polyhydric alcohol in the ophthalmic preparation, for example tyloxapol (tyloxapol) (0.1-1%), glycerol (0.2-1%), propylene glycol (0.2-1%), ethylene glycol (0.2-1%);
(b) polymerized polyalcohol, for example Polyethylene Glycol (for example PEG 300 and PEG 400)
(0.2-1%); (c) cellulose derivative (polymer of cellulose family), for example hydroxyethyl-cellulose (0.2-2.5%), hypromellose (0.2-2.5%), hydroxypropyl emthylcellulose (0.2-2.5%), methylcellulose (0.2-2.5%), sodium carboxymethyl cellulose (0.2-2.5%), hydroxypropyl cellulose (0.2-2.5%); (d) glucosan, for example macrodex (when using with another wetting agent, it is 0.1%); (e) water-solubility protein, for example gelatin (0.01%); (f) polyvinyl, for example polyvinyl alcohol (0.1-4%), polyethylene are than pyrrolidone (0.1-4%); (g) other polyhydric alcohol, for example polysorbate80 (0.2-1%), polyvidon (0.1-2%); (h) carbomer, for example carbomer 934 P, Carbopol 941, Acritamer 940 and carbomer 974P; And (i) polysaccharide/glycosaminoglycans, for example hyaluronic acid (hyaluronic acid/hyaluronate) (0.1-3%), chondroitin sulfate (0.1-3%).
Can in the present composition, add the viscosity that improves carrier more than a kind of viscosity intensifier.Preferred reinforcing agent in the carrier of peptide reagent preparation of the present invention is a carboxymethyl cellulose.
Viscosity
Viscosity has been described material after exerting pressure, and resists the internal force of mobile or alteration of form.The viscosity of material (solution, half viscogel, suspension, oils and fats ointment and soft gel (viscogel)) is with unit pool expression.Unit centipoise (" cp " or its plural form " cps ") equals 0.01 pool and uses through the pharmaceutical applications field of being everlasting.The chemical compound that is used for strengthening viscosity has multiple rank, as 15cps, 100cps etc.This number of levels is meant the viscosity that produces after the reinforcing agent of fixed percentage liquid solution is made.Usually, solution is 1% or 2%; Yet solution can be up to 4% in specific reinforcing agent.Measure viscosity at 20 or 25 ℃.
The appropriate viscosity of ophthalmic solution is between 25 to 50 centipoises (cps).Depend on the rank of reinforcing agent for the actual concentrations that produces the reinforcing agent that required viscosity needs.For example, if use methylcellulose 25cps, 1% solution will produce the viscosity of 25cps so.If use methylcellulose 4000cps, 0.25% solution can provide required viscosity so.Standard reference has provided the tabulation of the viscosity that percent solution and component rank produced.
According to the specific embodiment, preparation of the present invention also shows greater than 1 to 100,000 centipoise (cps) or higher viscosity.Ointment compositions of the present invention (grease gel or viscogel) can have greater than 100 the viscosity grade of 000cps.This be because ophthalmic ointment need thickness some, could stop it to flow away like this from the zone that needs are used.In time passing after application, these ointment are adhered in the conjunctival sac at place or the temperature of ocular surface can cause these ointment " fusing " and begins to flow.
The preferred viscosity ranges of multiple preparation type of the present invention sees the following form:
Preparation type range of viscosities (cps)
| Aqueous solution | ＞1 to＜1000 |
| Suspension | ＞1 to＜1000 |
| Gel | ＞3 to 40,000 |
| Ointment, gel | ＞20,000 to 100,000 |
| Ointment, oil | ＞20,000 to 100,000 |
Reducing agent/antioxidant/oxygen interleaving agent
Some compositions of the present invention has the possibility of oxidized degraded.Therefore; in the manufacturing process; the operation and the packing present composition can comprise: replace oxygen by (1) with nitrogen or high concentration noble gas such as argon, (2) add Reducing agent so that Oxidation minimizes, and (3) add bait molecule and protect chemical compound of the present invention to avoid the influence of Oxidation.
The concentration that can be used for common antioxidation (reduction) agent of ophthalmic preparation can reach 0.1% or higher, and they are sodium sulfate, sodium thiosulfite, sodium sulfite, sodium metabisulfite and thiourea.Sulphite can cause some anaphylaxiss; Therefore, before with the present composition treatment that contains this antioxidant, should inquire that the patient who takes this type of antioxidant is about this type of problem that may react.Other are applicable to that the available antioxidant of the present composition is ascorbic acid, EDTA/ disodiumedetate, acetic acid, citric acid, glutathion and acetylcysteine.These reagent also are regarded as stabilizing agent.
Bait molecule or oxygen insulation blocking reagent can be used as stabilizing agent to be added in the preparation of the present invention to minimize the Oxidation to preparation of the present invention.The molecule bait must have identical with preparation of the present invention at least oxidized ability.For the preparation of the present invention that contains methionine, such bait is exactly a methionine itself.The free methionine that is added in containing the reagent of the present invention of methionine can be competed oxygen in being oxidized to the process of methionyl.The free oxygen depleting agents is the oxidized reagent of oxygen activity aminoacid that stops in the present composition/peptide.For realizing the purpose of specific invention compositions, being not limited to the free oxygen depleting agents is methionine.
Ophthalmic ointment/oils and fats softens substrate
Ophthalmic ointment is easy to make the time of active agent and eye contact to be longer than suspension, and is also long than the time of solution certainly.The removal because difficult quilt is shed tears of most ointment is so be tending towards making the dimness of vision.Like this, ointment uses the auxiliary treatment as daily eye drop usually at night.
The oils and fats ointment base of the present composition is the mixture of mineral oil, vaseline and lanoline, and they all have the fusing point close with body temperature.Under chemical compound of the present invention, said composition can comprise mineral oil, vaseline or lanoline.According to the specific embodiment, preferred compositions comprises the combination of vaseline, mineral oil and lanoline.Most preferred compositions is the ointment compositions that contains white vaseline, mineral oil and lanoline (anhydrous).
The peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its conservative variant is dissolved in the saline of a spot of pure water or 0.9% to influence dissolution.This aqueous solution joins in the anhydrous lanolin, so then " water " lanoline (at the most 10%) and the softening matrix components of remaining ointment/oils and fats, mineral oil (at the most 30%) and white vaseline (at the most 60%) mixing mutually.
The ophthalmic ointment pipe contains the ointment of about 1-5 gram usually slightly, 3.5 grams preferably, and be furnished with thin head, and this thin head can be extruded ointment or its segment of inch meter slice, medicament is carried out quantitatively being used for.
Antiseptic
Aseptic is the absolute requirement of all ophthalmic preparations.The preparation that pollutes can cause eye infection, thereby finally causes losing one's sight, and especially, if contain Pseudomonas aeruginosa microorganism (P.aeruginosa), situation is then all the more so.Therefore, the ophthalmic preparation of describing herein must use the aseptic technology of solution, gel, suspension and ointment of guaranteeing the present composition.Sterile preparation must be packaged in the sterile chamber.Most external eyes with product usually with the multiple dose packaged.Therefore, need antiseptic to stop the microbial contamination of sterile product in the use.Suitable antiseptic comprises: quaternary ammonium compound (salt), as benzalkonium chloride (0.001-0.02%), hyamine 1622, cetalkonium chloride, cetrimonium bromide, benzene degree bromine ammonium and benzoxonium chloride; The alkyl mercuric salt of thiosalicylic acid, for example thimerosal (0.001-0.005%); Parabens, for example methyl parahydroxybenzoate and propyl p-hydroxybenzoate; Chelating agen is as disodiumedetate, gluconic acid sodium salt, sodium propionate; Other reagent, for example chlorobutanol, boric acid, sorbic acid, phenethanol (0.25%); Purite chlorine dioxide; Polyquad polyquatemium-1 (0.001%) and Aldox myristamide propyl group diethylamine (myristamidopropyl diethylamine) (0.005%) or other reagent well known by persons skilled in the art.
These antiseptic use the secondary microbial contamination that is caused by antibacterial, mycete and fungus with in guaranteeing to prevent to use with the level of 0.001%-0.1% (w/v) usually.
Below the selected Cmax of the antiseptic that can in ophthalmic preparation, use that now proved see the following form:
| Reagent | Cmax % |
| Benzalkonium chloride (BAK) hyamine 1622 methaform (clorobutanol) phenylmercuric acetate phenylmercuric nitrate thimerosal (thiomersal) methyl parahydroxybenzoate propyl p-hydroxybenzoate | ?0.01 ?0.01 ?0.5 ?0.004 ?0.004 ?0.01 ?0.2 ?0.04 |
Source: the OTC eye medicinal product of FDA advisory group, Final Report, in December, 1979
The selection of suitable preservatives is based on its antimicrobial efficient to selected invention compositions.The preferred preservative that is used for preparation of the present invention is combination, benzalkonium chloride (the BAK) (0.005%-0.02% of methyl parahydroxybenzoate (0.080%-1%) and propyl p-hydroxybenzoate (0.016%-0.024%), wherein 0.01%w/v is most preferred), the combination of BAK and EDTA (0.01-0.5%), they have synergism together when using.
Units dosage composition of the present invention is aseptic but not preservative treatment.Such compositions majority does not contain antiseptic.Therefore, these compositionss can not be used repeatedly, must abandon in case open.
Filling/stabilizing agent
Filling/stabilizing agent is useful to the hydration status of keeping lubricant, softening agent or carrier reinforcing agent in the present composition between the long-time storage life.Associate within the poly chain as if occur in these materials or between, the minimizing of these hydration status of having facilitated these chains in time of associating.These associations can be within the poly chain or between the hydrogen bond form, it can prove the variation of the viscosity and the quality of ophthalmic preparation/compositions of the present invention.Lyophilisation bulking agent mainly is a sugar, also can be regarded as the pressure protect agent, is used for protecting in freeze cycle chemical compound.The reagent that significantly lowers or eliminate the hydration status of this reduction is that the stable or hydration of a class strengthens reagent, that is, and and the polyhydric alcohol of 0.2-5wt% concentration.The representative of these polyhydric alcohol is mannitol, sorbitol, glycerol, sucrose, relevant sugar and analog.Most preferred stabilizing agent is the mannitol of suction, and its concentration is in the 0.2-5wt% scope.
In addition, the 50mM amino acid stabilizers, as alanine (Ala), lysine (Lys), glycine (Gly) and glutamic acid (Glu), also join and contain the making in the peptide formulations of sequence LKKTET or LKKTNT or its conservative variant to improve of the recovery of freezing back from recycled water solution.The preferred amino acids stabilizing agent is arginine and glycine, and most preferred 50mM aminoacid is glycine.
Using of peptide of the present invention
Exemplary topical administration (being used for the surface action effect)
For reaching partial result, the peptide formulations that will contain sequence LKKTET or LKKTNT or its conservative variant is administered to ocular surface, with treatment for example:
By, but be not limited to the corneal epithelium wound that causes by the epithelial debridement in chemical burn, recurrent corneal erosion, the operation, anterior corneal surface rebuilding operation, laser assisted in-situ keratomileusis (LASIK);
2. the corneal epithelium attenuation that causes by quaternary ammonium salt (as BAK and analog);
3. eye inflammation (using separately or with corticosteroid hormone), thus treatment is for example, conjunctivitis, blepharitis, keratitis, uveitis, scleritis, retinitis, optic neuritis and temporal arteritis;
4. treat infected by microbes (perhaps using separately) with antibacterium, antifungal or Anti-virus agent or with antimicrobial and anti-inflammatory agent;
5. dry eye syndrome (xerophthalmia);
6. blood-shot eye illness (separating congested agent (adrenal gland's energy vasoconstrictor of conjunctiva), for example ephedrine, naphazoline, phyenlephrinium, tetrahydrozoline and antihistaminic, for example pheniramine maleate separately or with eyes) makes together and is used for making eyes to bleach);
7. intraocular pressure (IOP) height and glaucoma; And
8. inflammation or the stimulation situation after traumatic injury or the surgical operation, or multiple eye stimulates inflammation or the stimulation situation in the disorder.
External peptide reagent of the present invention is made solution, suspension, gel and ointment.Peptide reagent preparation of the present invention can be direct or indirect pass through collagen sponge, insertion or similar methods use.The every kind of eye that comprises the external medicament for the eyes should be aseptic in its final container with product, to stop the infected by microbes to eyes.When preparation packing is used for repeatedly using in multi-dose container, keep aseptic for open the back at container, should in preparation, add antiseptic.Ophthalmic preparation needs careful its pH of control, buffer capacity, viscosity and osmotic pressure.Preferred pH scope, buffer agent, viscosity and osmotic pressure have been described herein.
Exemplary preparation: the externally used solution that is used for eye drop
The peptide formulations of the present invention of every milliliter of external comprises the following peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant:
The concentration range of peptide is about 0.001-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to reconcile NaOH or the HCI of pH; The USP purified water.
Exemplary preparation: the external suspension that is used for eye drop
Use suspension for the eye that comprises the peptide reagent that contains amino acid sequence LKKTET or LKKTNT, its granule must be less than 10 microns to minimize the stimulation to eyes.Solid not dissolved particles tends to adhere on the conjunctiva.After medicine is absorbed, thereby these granules can dissolve and replenish the medicine be absorbed.Compare with solution, this accumulate or the storage effect improved suspension time of contact and the effect duration.
The peptide formulations of the present invention of every milliliter of external comprises the following peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant:
The concentration range of peptide is about 0.00 1-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): the peptide encapsulation of poly-(lactide-Acetic acid, hydroxy-, bimol. cyclic ester) PLGA microspheres form; The 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to regulate NaOH or the HCI of pH; The USP purified water.
Exemplary preparation: the external-use gel that is used for eye drop
The peptide formulations of the present invention of every milliliter of external comprises the following peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant:
The concentration range of peptide is about 0.001-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): sodium carboxymethyl cellulose (0.5-1%); Disodium hydrogen phosphate; Sodium chloride; Propylene glycol; Methyl parahydroxybenzoate; Propyl p-hydroxybenzoate; Be used to regulate NaOH or the HCI of pH; The USP purified water.
Exemplary preparation: externally-applied ointment
The peptide formulations of the present invention of every milliliter of external comprises the following peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant:
The concentration range of peptide is about 0.001-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle) (1): sodium carboxymethyl cellulose (2.5%); Disodium hydrogen phosphate; Propylene glycol; Methyl parahydroxybenzoate; Propyl p-hydroxybenzoate; Sodium chloride; Be used to regulate NaOH or the HCI of pH; The USP purified water.
Preferred carrier (carrier/vehicle) (2): " water " lanoline (10%); Mineral oil (30%) and white vaseline (60%).
Save the exemplary topical administration of steroid effect
Corticosteroid has suppressed the anti-inflammatory response to multiple zest reagent.
● dexamethasone eye suspending agent (0.1%); Dexamethasone ophthalmic ointment (0.05%);
With dexamethasone sodium phosphate ophthalmic solution (0.1%)
● fluorometholone ophthalmic ointment (0.1%); Fluorometholone eye suspension (0.25-1%); With fluorometholone acetate eye suspension (0.1%)
● loteprednol (0.5%)
● medrysone eye suspension (1%)
● the prednisoni acetas Arillus Longan is used suspension (0.12-1%) and prednisolone phosphate sodium ophthalmic solution (0.125-1%)
● rimexolone eye suspension (1%)
Yet these reagent can improve intraocular pressure (IOP), thereby can induce the glaucoma of susceptible individual to damage optic nerve, make the vision accuracy and the visual field is impaired, form posterior subcapsular cataract.Cataractous formation more seemingly with high dose, the long-time complication together of using.Some corticosteroid, for example fluorometholone acetate, medrysone and loteprednol cause that than other corticosteroid less IOP raises.Thereby life-time service also can suppress host's immunoreation to have helped to form the secondary ocular infection from fungus and virus that ocular tissue discharges.Known external corticosteroid delays or the healing of the wound that slows down.
Use the external eye drop or the ointment that comprise the peptide reagent of the present invention that contains amino acid sequence LKKTET or LKKTNT or conservative variant and suppress that the inflammatory reaction that stimulates reagent is had the potential of saving steroid.
Exemplary eye drops-conjunctiva/sclera instils
External conjunctiva route of entry goes in the anterior ocular segment to play an important role at drug osmotic.And, found that external used medicine enters sclera from conjunctiva.Probability by sclera transhipment or diffusion depend on the big and permeable surf zone of this tissue, with and not with significantly reduced hyperhydrated of age, height hypocellularity and high-permeability.Therefore, can expect that compositions of the present invention can find the path that arrives anterior ocular segment by this non-intruding route of administration.The data declaration sclera in addition to the chemical compound of macromolecule (～150kD) also be easy to penetrate, this is much larger than the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its conservative variant.Up-to-date discovery: the external nepafenac suppresses choroid by the generation that reduces VEGF and amphiblestroid new vessels forms, macromolecule peptide such as insulin (5.8kD) can be in retina and the accumulation of optic nerve place after the external administration, this explanation external compositions of the present invention, its molecular weight all＜150kD, can not only pass conjunctiva to anterior ocular segment, and they also has therapeutical effect.Externally used solution, suspension, gel or the ointment of the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant of Miao Shuing are the outer appropriate formulation that is used for conjunctivae and selerae hereinbefore.
In addition, the present composition that contains the peptide reagent of amino acid sequence LKKTET or LKKTNT or its conservative variant by injection carries out using under the conjunctiva and can be used for sending anti-inflammatory and antimicrobial therapy (and be responsive to chemical compound of the present invention), can be used for treating serious eye inflammation and ocular infection, for example uveitis, endophthalmitis and glaucoma.
Preferred injectable formulation: the peptide reagent that every ml contains amino acid sequence LKKTET or LKKTNT or conservative variant comprises:
The concentration range of peptide is at about 0.001-1,000mg/ml, and preferred at about 0.1-60mg/ml preferably at about 0.01-600mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to regulate NaOH or the HCI of pH; The USP purified water.
Exemplary eye drops-saturating cornea instils
External used medicine is by the pleasing to the eye interior environment of hydrophobic corneal osmosis; Yet saturating cornea transhipment is not effective and efficient manner, and reason is that it makes 1/10th to 3/10ths dosage infiltrate through eyes and most drug remains in the surface of epithelial layer.Peptide combinations of the present invention is by the passive influence that is diffused in the dissolubility, molecular weight and the degree of ionization that are subjected to them to a great extent of cornea.Make the peptide reagent of the present invention of external used medicine owing to have clean negative electricity and relative high molecular weight, thereby be difficult to penetrate complete cornea.The fact that hole between corneal epithelial cell only allows about 500 molecular weight or littler micromolecule cell bypass to pass is supported this viewpoint.Yet, when complete cornea be corroded when destroying, for example or be exposed to and can open between epithelial cell close-connected certain material or penetrate reagent, compositions of the present invention can more effectively be passed cornea and be entered the ophthalmic space.
Externally used solution, gel or the ointment of the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or conservative variant as indicated above is the appropriate formulation that cornea instils.
Exemplary eye drops-periocular injections
Do not reply independent external eye drop and under inflammatory conditions such as anterior uveitis, posterior uveitis, endophthalmitis and optic neuritis situation, use the periocular injections preparation of peptide reagent of the present invention at eye inflammation.This peptide reagent is injected under the conjunctiva or fascial bursa (Tenon ' s capsule) under the space.Thereby absorbing can be more, and the result is that desired target spot has more medicine.Periocular injections is replenishing of external curing, but it is inconvenient and be not suitable for first-selected treatment.
Preferred injectable formulation: the peptide reagent that every ml contains amino acid sequence LKKTET or LKKTNT or conservative variant comprises:
The concentration range of peptide is about 0.001-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to regulate NaOH or the HCI of pH; The USP purified water.
In exemplary eye drops-vitreous body/aqueous humor in (IntraAqueous) use
As substituting of saturating cornea, saturating conjunctiva and saturating sclera transhipment, the eye inner tissue that sends into of peptide reagent of the present invention can be by being injected into the realization of vitreous body or aqueous humor intracavity.Vitreous body is to be made of hydrogel (water, hyaluronic acid and collagen), and it has filled the chamber between retina and the crystalline lens, and aqueous humor is the aqueous fluids of filling chamber between crystalline lens and the iris.The peptide reagent of the present invention of making solution is expelled to makes eye inner tissue be exposed to peptide reagent rapidly in vitreous body and the aqueous humor.Reagent can be disposed from vitreous body fast, and institute thinks that realization reagent of the present invention continues to exist within the eye, just need duplicate injection, and duplicate injection has improved the risk of suffering from endophthalmitis, damaging crystalline lens and cause retina shedding, and has been difficult to stand.For overcoming this obstacle; peptide reagent of the present invention can be wrapping in the immobilized artificial membrane; be that liposome, biodegradable microsphere, nano-particle or biodegradable lactone are the polymer of substrate, it comprises the polyester of being made by L-lactide, Acetic acid, hydroxy-, bimol. cyclic ester, caprolactone, dioxanone, cyclic carbonate and their derivant polycondensation.Polyactide and poly-Acetic acid, hydroxy-, bimol. cyclic ester also are called poly-(lactic acid) PLA and poly-(glycolic) PGA, especially their copolymer polylactic acid/ethanol copolymer PLGA are the maximum biodegradable polymers of research, and it also can be used as the carrier of peptide reagent of the present invention.In addition, the peptide reagent and synthetic reach natural polymer such as Polyethylene Glycol (PEG) and glucosan (comprise ring glucosan) covalently bound of the conjugate of peptide-polymer as containing LKKTET or LKKTNT or its variant, can improve the pharmacokinetics pattern, thereby cause having reduced removing peptide of the present invention.
Using peptide reagent of the present invention in the vitreous body or in the aqueous humor can be in eye inflammation, ocular infection (antibacterial, fungus or virus) and glaucomatous treatment, structure by F-actin in the control outflow pathway cell represents (Read AT et al., Exp Eye Res, 2006Jun:82 (6): 974-85).
Preferred injectable formulation: the peptide reagent that every ml contains amino acid sequence LKKTET or LKKTNT or conservative variant comprises:
The concentration range of peptide is about 0.001-1,000mg/ml, and preferably about 0.01-600mg/ml, more preferably about 0.1-60mg/ml, most preferred peptide concentration is about 1-6mg/ml.
Preferred carrier (carrier/vehicle): 1): the 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to regulate NaOH or the HCI of pH; The USP purified water.
Preferred carrier (carrier/vehicle) (2): the peptide encapsulation of PLGA microspheres form; The 20mM sodium citrate; The 50mM glycine; 3% sucrose; Be used to regulate NaOH or the HCI of pH; The USP purified water.。
The exemplary formulation dosage
Externally used solution and suspension
Suggestion is used one at every turn and was also used once more in 5 minutes at least at interval.In eyes, instil and oppress lachrymal sac 1-2 minute immediately to reduce drug wastage speed after one by this path.Injectable dosage: use the pin of 27-30 specification, 0.5 inchage.
Compositions according to the specific embodiment can be a lyophilized form, perhaps can be by freeze dried form, it comprises the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant, perhaps comprise the stimulation reagent that stimulates LKKTET or LKKTNT peptide or its conservative variant to produce, said composition further comprises at least a amino acid stabilizers.Said composition can comprise the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant, perhaps comprise the stimulation reagent that stimulates LKKTET or LKKTNT peptide or its conservative variant to produce, and at least a in lyophilizing filler or the amino acid stabilizers, described compositions is in lyophilized form.Said composition further comprises a kind of in acidity or the alkaline pH regulator, and this pH adjusts the pH that can regulate compositions in water-bearing media and can accept the pH level to required physiology, and in described water-bearing media the buffer agent of basicly stable above-mentioned required pH.Amino acid stabilizers can comprise at least a in alanine, lysine, glycine or the glutamic acid.Amino acid stabilizers can comprise the amino acid stabilizers of at least a 50mM.Amino acid stabilizers can comprise the 50mM glycine.Said composition can further comprise filling reagent, and this filling reagent comprises at least a in carbohydrate, sugar alcohol, monosaccharide, disaccharide, polysaccharide, polyhydric alcohol, mannitol, sorbitol, glycerol, sucrose or the glucose dextrose.The pH regulator agent can comprise at least a among NaOH or the HCl.Buffer agent can comprise sodium salt at least a in acetic acid, ascorbic acid, boric acid, carbonic acid, phosphoric acid, citric acid, gluconic acid, lactic acid or the propanoic acid, the calcium salt of carbonic acid or propanoic acid, potassium phosphate, the Tris buffer agent, acetic acid, phosphoric acid, at least a in citrate or the borate buffer.Buffer agent can be a sodium citrate.Buffering agents is about 0.05-1.0.Buffering agents is about 0.02-0.2, or about 0.01-0.1.The scope of required pH level can be about 3.5-11.5, about 3.5-9, approximately 4.5-8, or about 5.0-7.8.Required pH level can be about 5.5.Peptide reagent comprises the N-end variable body of aminoacid sequence KLKKTET, amino acid sequence LKKTET Q, T β 4, T β 4, the C-end variable body of T β 4 or the isomer of T β 4.Compositions can further comprise water-bearing media, and the concentration range of wherein said peptide reagent in described water-bearing media is about 0.001-1,000mg/ml.Compositions can further comprise at least a steroid.
Compositions according to another specific embodiment is used for experimenter's skin histology is used, and comprise the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant, perhaps be included in the described stimulation reagent of organizing internal stimulus LKKTET or LKKTNT peptide or its conservative variant to produce, said composition further comprises quaternary ammonium salt and is applied to the external carrier of described experimenter's skin histology.This peptide reagent can comprise the isomer of N-end variable body or the T β 4 of amino acid sequence LKKTET, amino acid sequence LKKTET Q, T β 4, T β 4.Quaternary ammonium salt can comprise benzalkonium chloride.The concentration of peptide reagent can be greater than 0.001-1, and 000mg/ml, described quaternary ammonium salt approximately are 0.001-1wt% in described compositions.Said composition can be solution, gel, emulsifiable paste, paste, lotion, spray, suspension, dispersion liquid, ointment, the form of hydrogel, ointment or foam formulations.Said composition can be a cosmetic formulations.
According to the pharmacy of another specific embodiment or cosmetics and use medicine, it comprises the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant, perhaps comprise the stimulation reagent that stimulates LKKTET or LKKTNT peptide or its conservative variant to produce, said composition further comprises quaternary ammonium salt, and wherein said reagent and described salt can be used the experimenter separately or together.This peptide reagent can comprise the isomer of N-end variable body or the T β 4 of amino acid sequence LKKTET, amino acid sequence LKKTET Q, T β 4, T β 4.Quaternary ammonium salt can comprise benzalkonium chloride.Should and can comprise pharmacy, eye usefulness or cosmetic composition with medicine, the concentration of its contained peptide reagent can be greater than 0.001-1, and 000mg/ml, wherein said quaternary ammonium salt approximately are 0.001-1wt% in described compositions.Should and can comprise ophthalmic composition with medicine, this ophthalmic composition further comprises can use carrier.Said composition can comprise the eye drop compositions.
According to another specific embodiment, treatment, stop, suppress or reduce tissue owing to the experimenter is used decline, processing method impaired or infringement that quaternary ammonium salt causes comprise described receptor is used described quaternary ammonium salt and described experimenter is used the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant or at the described stimulation reagent of organizing internal stimulus LKKTET or LKKTNT peptide or its conservative variant to produce.This reagent can be before described quaternary ammonium salt be used, simultaneously or afterwards the experimenter is used.This reagent and described salt can be used as compositions and use simultaneously.Said composition can further comprise can the eye carrier.Said composition can comprise the eye drop compositions.Said composition can comprise the cosmetics acceptable carrier.Said composition can be solution, gel, emulsifiable paste, paste, lotion, spray, suspension, dispersion liquid, ointment, the form of hydrogel, ointment or foam formulations.Peptide reagent can comprise the N-end variable body of aminoacid sequence KLKKTET, amino acid sequence LKKTET Q, T β 4, T β 4, the C-end variable body of T β 4 or the isomer of T β 4.Quaternary ammonium salt can comprise benzalkonium chloride.The concentration that is included in the peptide reagent in the described compositions can be about 0.001-1, and 000mg/ml, described quaternary ammonium salt approximately are 0.001-1wt% in described compositions.
According to the compositions of another specific embodiment, it comprises the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its variant or stimulates LKKTET or stimulation reagent that LKKTNT peptide or its conservative variant produce and can eye high osmotic agent, the comfort enhancers of eye with osmotic pressure are provided, can regulate compositions pH to required the buffer agent of using at least a in the acidity that can accept the pH level or the alkaline pH regulator and keeping above-mentioned required pH level substantially with the antiseptic of carrier, anti-microbial effect, for described compositions.Said composition can comprise further that antioxidant or oxygen isolates a kind of in the reagent.Antioxidant or oxygen are isolated reagent can comprise sodium sulfite, sulfo-pressure sodium sulfate, sodium sulfite, pyrosulfurous acid hydrogen sodium, thiourea, ascorbic acid, EDTA/ disodiumedetate, acetic acid, citric acid, glutathion, acetylcysteine or methionine.It is about 0.0001-1.0wt% that antioxidant or oxygen are isolated the concentration range of reagent in described compositions.The antiseptic of anti-microbial effect can comprise a kind of in following: quaternary ammonium compound: benzalkonium chloride, hyamine 1622, cetalkonium chloride, cetrimonium bromide, benzene degree bromine ammonium, benzoxonium chloride; The alkyl mercuric salt of thiosalicylic acid: thimerosal, phenylmercuric nitrate, phenylmercuric acetate, phenylmercuric borate; Parabens: methyl parahydroxybenzoate, propyl p-hydroxybenzoate; Chelating agen, disodiumedetate, gluconic acid sodium salt, sodium propionate; Alcohols, chlorobutanol, benzyl alcohol, phenethanol; Guanidine derivatives: chlorhexidine (chlorohexidine), polyhexamethylene biguanide (polyhexamethylene biguanide), sorbic acid, boric acid, chlorine dioxide, polyimidazole hyamine (polyquatemium) or myristamide propyl group diethylamine (myristamidopropyldiethylamine).The concentration range of antiseptic in described compositions is about 0.0001-5.0% (w/v).Said composition can comprise further that at least a eye is with accepting stabilizing agent.Stabilizing agent can comprise at least a in polyhydric alcohol, mannitol, sorbitol, glycerol, sucrose, amino acid stabilizers alanine (Ala), lysine (Lys) or the glutamic acid (Glu).The concentration range of stabilizing agent in described compositions is about 0.01-10wt%.High osmotic agent comprises ionic compound, alkali metal halides, alkaline-earth metal halides, CaCl
2, at least a in KBr, KCl, LiCl, NaBr, NaCl, boric acid, non-ionic compound, carbamide, glycerol, sorbitol, mannitol, propylene glycol or the glucose dextrose.The scope of the osmotic pressure of said composition can be 50-1000mOsmol.Regulator can comprise at least a among NaOH or the HCl.Buffer agent can comprise at least a in acetic acid sodium salt, SODIUM ASCORBATE, boric acid sodium salt, sodium carbonate salt, sodium ascorbyl phosphate, sodium citrate salt, gluconic acid sodium salt, lactylate salt, propionic acid sodium salt, polcarb, propanoic acid calcium salt, potassium phosphate, Tris buffer agent, acetate buffer agent, phosphoric acid buffer agent, citrate buffer agent or the borate buffer.The scope of required pH level is about 3.5-11.5.The buffer capacity that buffer agent has is about 0.05-1.0.Peptide reagent can comprise the N-end variable body of aminoacid sequence KLKKTET, amino acid sequence LKKTET Q, T β 4, T β 4, the C-end variable body of T β 4 or the isomer of T β 4.This chemical compound further comprises aqueous medium, and the concentration range of wherein said peptide reagent in described water-bearing media is 0.001-1,000mg/ml.Said composition can be solution, gel, emulsifiable paste, paste, lotion, spray, suspension, dispersion liquid, ointment, the form of hydrogel, ointment or foam formulations.Said composition can comprise the eye drop compositions.Peptide reagent can be in described compositions, and compositions is by at least a parcel the in immobilized artificial membrane, liposome, microsphere, nano-particle or the biodegradable polymer, and perhaps the form of the conjugate of peptide-polymer exists.
Embodiment
Extrasin beta 4 (T β 4) is one and contains 43 amino acid whose molecules that the healing that it has promoted the eye wound has reduced eye inflammation, and corneal epithelial has the anti-apoptotic effect.In this research, detected the effect of survival that 4 couples of T β are exposed to the cultivation people corneal epithelial cell of benzalkonium chloride (BAK).
The about 80% people's corneal epithelial cell that converges was handled 15 minutes at the BAK with 0%, 0.001%, 0.01% or 0.1%.In culture medium, recover after 3 and 24 hours, use colorimetric BrdU to infiltrate analysis and detect the cell increment.Use the colorimetric annexin to detect apoptosis as the cell death analysis on basis.Repeated experiments in 1mcg/ml T β 4, this dosage show in several researchs of having delivered effectively.Stop apoptotic ability for further assessing T β 4, corneal epithelial cell is handled above 5 days with 0.01%BAK ± T β 4.
Under all used BAK concentration, compared with the control, corneal epithelial cell has been suppressed in the propagation of 3 and 24 hours recovery time, and apoptosis has been enhanced.Time point at 3 and 24 hours, T β 4 does not eliminate the illeffects of BAK; T β 4 does not promote that cell proliferation and apoptosis are not suppressed yet.Yet when longer incubation time (2-5 days), T β 4 handles and has significantly suppressed the inductive epithelial cell apoptosis of BAK.In addition, compare with the cell of only cultivating 5 days in culture medium, the apoptosis of the cell that T β 4 handles has reduced.
BAK is the antiseptic that uses in many commercially available ophthalmic solutions, and it has induced epithelial apoptosis in cultivation, illustrates that it is deleterious exposing corneal health for a long time.Harmful apoptosis previous crops that the research explanation T β 4 of this paper report can overcome BAK is used.Because the eye drop of many BAK of containing uses in a very long time usually, so T β 4 is the useful additives that contain the solution of this kind antiseptic.
Claims (20)
1. compositions, it comprises the peptide reagent that contains amino acid sequence LKKTET or LKKTNT or its conservative variant or stimulates LKKTET or stimulation reagent that LKKTNT peptide or its conservative variant produce, and eye with carrier, effectively anti-microbial preservative, for described compositions provide eye with high osmotic agent, the comfort enhancers that can accept osmotic pressure, can regulate compositions pH to required with at least a in the acidity of pH level or the alkaline pH regulator with keep the buffer agent of above-mentioned required pH level substantially.
2. compositions according to claim 1, it further comprises at least a in antioxidant or the oxygen interleaving agent.
3. compositions according to claim 2, wherein said antioxidant or oxygen interleaving agent comprise at least a in sodium sulfite, sodium thiosulfite, sodium sulfite, pyrosulfurous acid hydrogen sodium, thiourea, ascorbic acid, EDTA/ disodiumedetate, acetic acid, citric acid, glutathion, acetylcysteine or the methionine.
4. compositions according to claim 3, wherein said antioxidant or the concentration range of oxygen interleaving agent in described compositions are about 0.0001-1.0wt%.
5. compositions according to claim 1, wherein said effective antiseptic comprise at least a in the following chemical compound: quarternary ammonium salt compound: benzalkonium chloride, hyamine 1622, cetalkonium chloride, cetrimonium bromide, benzene degree bromine ammonium, benzoxonium chloride; The alkyl mercuric salt of thiosalicylic acid: thimerosal, phenylmercuric nitrate, phenylmercuric acetate, phenylmercuric borate; P-Hydroxybenzoate: methyl parahydroxybenzoate, propyl p-hydroxybenzoate; Chelating agen, disodiumedetate, gluconic acid sodium salt, sodium propionate; Alcohol: chlorobutanol, benzyl alcohol, phenethanol; Guanidine derivatives: chlorhexidine, polyhexamethylene biguanide, sorbic acid, boric acid, chlorine dioxide, polyimidazole hyamine or myristamide propyl group diethylamine.
6. compositions according to claim 5, the concentration range of wherein said antiseptic in described compositions are about 0.0001-5.0% (w/v).
7. compositions according to claim 1, it further comprises at least a eye stabilizing agent.
8. compositions according to claim 7, wherein said stabilizing agent comprise at least a in following: polyhydric alcohol: mannitol, sorbitol, glycerol, sucrose; Amino acid stabilizers: alanine (Ala), lysine (Lys) or glutamic acid (Glu).
9. compositions according to claim 8, the concentration range of wherein said stabilizing agent in described compositions is about 0.01-10wt%.
10. compositions according to claim 1, wherein said high osmotic agent comprise at least a in following: ionic compound: alkali metal halides, alkaline-earth metal halides, CaCl
2, KBr, KCl, LiCl, NaBr, NaCl, boric acid; Non-ionic compound: carbamide, glycerol, sorbitol, mannitol, propylene glycol or glucose dextrose.
11. compositions according to claim 10, the scope of the osmotic pressure that it had are about 50-1000mOsmol.
12. compositions according to claim 1, wherein said pH regulator agent comprise at least a among NaOH or the HCl.
13. compositions according to claim 1, wherein said buffer agent comprise at least a in acetic acid sodium salt, SODIUM ASCORBATE, boric acid sodium salt, sodium carbonate salt, sodium ascorbyl phosphate, sodium citrate salt, gluconic acid sodium salt, lactylate salt, propionic acid sodium salt, polcarb, propanoic acid calcium salt, potassium phosphate, Tris buffer agent, acetate buffer, phosphate buffer, citrate buffer agent or the borate buffer.
14. compositions according to claim 1, the scope of wherein said required pH level is about 3.5-11.5.
15. compositions according to claim 1, wherein said buffering agents are about 0.05-1.0.
16. compositions according to claim 1, wherein said peptide reagent comprise the N-end variable body of aminoacid sequence KLKKTET, amino acid sequence LKKTET Q, T β 4, T β 4, the C-end variable body of T β 4 or the isomer of T β 4.
17. compositions according to claim 1, it further comprises water-bearing media, and the concentration range of wherein said peptide reagent in described water-bearing media is about 0.001-1,000mg/ml.
18. compositions according to claim 1, wherein said compositions are the forms of solution, gel, emulsifiable paste, paste, lotion, spray, suspension, dispersion liquid, ointment, hydrogel, ointment or foam formulations.
19. compositions according to claim 1, wherein said compositions comprises the eye drop compositions.
20. compositions according to claim 1; wherein said peptide reagent is present in the described compositions; this peptide reagent is by at least a parcel the in immobilized artificial membrane, liposome, microsphere, nano-particle or the biodegradable polymer, and perhaps the conjugate form with peptide-polymer exists.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US69126105P | 2005-06-17 | 2005-06-17 | |
| US60/691,261 | 2005-06-17 | ||
| US60/776,947 | 2006-02-28 |
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| Publication Number | Publication Date |
|---|---|
| CN101198346A true CN101198346A (en) | 2008-06-11 |
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ID=39548325
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006800216620A Pending CN101198344A (en) | 2005-06-17 | 2006-06-19 | LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized |
| CNA2006800216616A Pending CN101198346A (en) | 2005-06-17 | 2006-06-19 | LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized |
| CNA2006800216692A Pending CN101198345A (en) | 2005-06-17 | 2006-06-19 | LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006800216620A Pending CN101198344A (en) | 2005-06-17 | 2006-06-19 | LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006800216692A Pending CN101198345A (en) | 2005-06-17 | 2006-06-19 | LKKTET and/or LKKTNT peptide compositions which are lyophilized or in a form capable of being lyophilized |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110430866A (en) * | 2017-03-03 | 2019-11-08 | Gtreebnt科技有限公司 | Stabilization external preparation containing extrasin beta -4 as effective component |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104271142B (en) * | 2012-03-01 | 2017-07-04 | 福斯特斯特林研究公司 | The topical gel of PROTEIN C end (ACT) peptide is connected containing α |
| CN102924573B (en) * | 2012-11-13 | 2014-07-23 | 兆科药业（广州）有限公司 | Actin binding peptide and purpose thereof |
| KR102488796B1 (en) * | 2016-07-18 | 2023-01-13 | 리젠트리 엘엘씨 | How to treat dry eye syndrome |
| CN111712229A (en) * | 2017-09-15 | 2020-09-25 | 奥叙拉尔有限公司 | Ophthalmic delivery device |
| CN115266995B (en) * | 2022-08-02 | 2023-10-13 | 兆科（广州）眼科药物有限公司 | Analysis method of related substances in pilatory peptide preparation |
-
2006
- 2006-06-19 CN CNA2006800216620A patent/CN101198344A/en active Pending
- 2006-06-19 CN CNA2006800216616A patent/CN101198346A/en active Pending
- 2006-06-19 CN CNA2006800216692A patent/CN101198345A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110430866A (en) * | 2017-03-03 | 2019-11-08 | Gtreebnt科技有限公司 | Stabilization external preparation containing extrasin beta -4 as effective component |
| CN110430866B (en) * | 2017-03-03 | 2023-04-07 | Hlb医疗有限公司 | Stable external preparation containing thymosin beta-4 as active ingredient |
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| CN101198344A (en) | 2008-06-11 |
| CN101198345A (en) | 2008-06-11 |
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