CN103142462A

CN103142462A - Brinzolamide eye preparations, and preparation method and use thereof

Info

Publication number: CN103142462A
Application number: CN2013100553679A
Authority: CN
Inventors: 宋相容; 魏于全
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2012-02-23
Filing date: 2013-02-21
Publication date: 2013-06-12
Anticipated expiration: 2033-02-21
Also published as: CN104814924A; CN103142462B

Abstract

The invention relates to the medicine field, and concretely relates to brinzolamide eye preparations, and preparation methods and uses thereof. Several brinzolamide eye preparations are provided to prolong the eye stay time or improve the bioavailability, and provide a new dose form choice for the reduction of the intraocular tension or the treatment of glaucoma. The concrete dose forms of the preparations comprise a temperature-sensitive eye in-situ gel, a pH-sensitive eye in-situ gel, an ion-sensitive eye in-situ gel, a brinzolamide lipidosome, a brinzolamide nanoparticle, a brinzolamide nano-suspension, and a brinzolamide composite gel preparation, the preparations can be used for treating diseases comprising intraocular hypertension, glaucoma and the like, and the gels can prolong the stay time of medicines in eyes and reduce the administration frequency and the administration dosage, so the patient compliance is improved, and the side effects of medicines are reduced. The no addition of any antiseptics and the adoption of single-dosage packaging reduce the irradiation of the use of eye drops for a long term to eyes and substantially improve the medicine safety.

Description

Brinzolamide ophthalmic preparation and its production and use

Technical field

The present invention relates to field of medicaments, be specifically related to brinzolamide ophthalmic preparation and its production and use.

Background technology

Glaucoma is a kind of disease that causes optic nerve lesion.Optic nerve is comprised of a lot of nerve fibers, when increased intraocular pressure, can cause nerve fiber damage, causes defect of visual field.Control glaucomatous key and be and reduce intraocular pressure, and the generation of aqueous humor, to discharge be the key of controlling intraocular pressure, unbalance if aqueous humor generates, discharges, and causes that wall of eyeball pressure is too large, causes optic nerve lesion.Carbonic anhydrase (CA) is present in a lot of bodily tissues that comprise ocular tissue, and catalysis carbon dioxide aquation becomes carbonic acid, and the reversible reaction of carbonic acid dehydration.Corpus ciliare produces aqueous humor by this reaction, thus suppress the secretion that the capsulociliary carbonic anhydrase of eye can reduce aqueous humor, thus reduce intraocular pressure.Brinzolamide mainly suppresses dominant carbonic anhydrase II type isozyme in ocular tissue, thereby reduces the generation of aqueous humor, is applicable to ocular hypertension, the high intraocular pressure of open-angle and the invalid glaucoma of other Drug therapys.

EP2348026 and EP1985618 disclose the synthetic of brinzolamide, US5461081, US5378703, US5240923 and US005948801A disclose the synthetic of brinzolamide and as the use of eye drop, the prescription that suspensoid is wherein arranged, be the Pai Liming eye drop (Azopt) of Alcon Universal Ltd., its preparation is suspensoid, can reach the effect of slow release, but patient tolerability is not good, and combine with other drug while using and can only make suspensoid, its application is had to very large limitation; JP2010037327 discloses the patent of brinzolamide water formulation, but, after the medicine salify, its eye holdup time is short, and the cornea permeability is not good, and slow releasing function is poor; WO2009073216A1 proposes the prescription of brinzolamide nano suspension, but need to make lyophilized formulations, and concerning the glaucoma patient that needs medication every day, it is very inconvenient to use; CN2005100434124, its gel-type vehicle is selected from the multivalence hydrochlorate of methylcellulose, is the cavity thermosensitive hydrogel, but does not propose the detailed prescription of brinzolamide preparation; CN2010101485162 discloses the method that high-pressure electrostatic prepares chitosan gel rubber, and its gel-type vehicle is selected from chitosan, but propose brinzolamide, specifically do not write out a prescription.CN2010100011309 discloses a kind of temperature sensitive gel for eye use, and its gel-type vehicle is chitosan etc., its objective is and extends the retention time of preparation in eye, also without the detailed prescription of brinzolamide.In a word, the said goods and patent all do not solve the zest problem of brinzolamide eye drop short in the eye retention time in antiseptic life-time service simultaneously, this area be starved of by brinzolamide make safety, efficient, stable, the controlled eye that facilitates drips the liquid preparation of use.

Gel for eye use often be take hydrophilic high molecular material as carrier, biocompatibility is preferably arranged, and is semi-solid preparation, but the action time of prolong drug, reduce administration number of times, can overcome that the eye drop bioavailability is low, the shortcoming such as the greasy not easy to apply and suspensoid ophthalmic uncomfortable of Eye ointments.And the specific physico chemical factor of situ-gel system in conjunctival sac (such as: pH, temperature, ion etc.) under, self will produce the phase transformation of sol-gel, and rarer liquid rotating becomes thicker gel when splashing into, because causing mutually the difference of the thixotropic factor changed, be divided into again three kinds of pH responsive type, responsive to temperature type and ion-sensitive types.The invention human desires is developed the in-situ gel of brinzolamide to reach the long-acting control intraocular pressure of slow releasing function, and, for the use in conjunction with other intraocular pressure lowering medicines provides probability, improves patient's compliance.

Eye is medicine to be embedded in to the cavity of cyclodextrin with clathrate, cyclodextrin carries medicine and is scattered in solvent, the solubilization of realization to medicine, and in certain scope, the ratio that increases cyclodextrin is conducive to improve the drug accumulation corneal permeability, therefore the inventor intends using cyclodextrin to carry out enclose to medicine, preparation has the lyophilized formulations of good cornea permeability.

People's corneal epithelium is lipid layer, the difficult cornea barrier that sees through of hydrophilic medicine, and although lipophilic drugs can pass through cornea, but its dissolubility in preparation is not high, the Chang Weinan soluble substance, as brinzolamide, therefore medicine will see through cornea and reach treatment concentration, just need more suitable lipotropy and hydrophilic, the lipophilic hydrophilic of chemical substance usually characterizes by Determination of oil-water partition coefficient, Determination of oil-water partition coefficient relatively is suitable for seeing through cornea barrier arrival focus at 2 to 3 medicine or complex, the n-octyl alcohol water partition coefficient physiological pH of brinzolamide is 6.6, pH is within 5 o'clock, to be 0.7, and after its preparation is become to liposome, Determination of oil-water partition coefficient raises, can effectively reduce the effective dose of medicine, therefore the inventor intends using the brinzolamide liposome as a kind of scheme that solves its slightly solubility.

The inventor also intends using the nano material such as PLGA to prepare the brinzolamide nanoparticle simultaneously, and the medicine parcel is become to nanometer particle, existence that can be stable in aqueous solution, and increased its dissolubility in water, make it there is drug effect preferably.

Nano suspension is not use any carrier, the submicron dispersion system of colloid formed in disperse medium by pure drug particle high degree of dispersion under a small amount of stabilizer function, and particle diameter is generally at 10～1000nm.Its specific surface area is large, and medicine is stripping rapidly, effectively solves the problem that drug solubility is low.At present, the technology for preparing nano suspension has wet grinding, the sedimentation method, microjet method, high pressure homogenization method.High pressure homogenization method is to make the thick suspension that contains medicine, stabilizing agent at a high speed by a very narrow gap, and the huge energy produced due to cavitation, high speed shear power and interparticle collision makes drug particle be pulverized as nanoparticle.High pressure homogenization method is compared with additive method, operate short, favorable reproducibility comparatively simple, consuming time, can facilitate to the particle diameter of product adjusting by regulating homogenization pressure and cycle-index, obtain the nano suspension of different large small particle diameter: equipment cost is low, and technology is comparatively ripe, has been approved in the commercial production of intravenous injection product lipomul; The products obtained therefrom particle diameter is little, and and even, physical and chemical stability is good, applied widely.Nano suspension belongs to the submicron system of high degree of dispersion, has higher surface free energy, is a kind of thermodynamic instability system.Under liquid condition, store, its particle is easily assembled, adhesion, and to reduce the surface free energy of system, common processing method is to add freeze drying protectant, by Freeze Drying Technique, makes stable freeze-dried powder.

As mentioned before, the various brinzolamide eye drip preparations that the invention human desires is implemented have characteristics separately, can make ophthalmic preparation by some specific methods, if can be by the brinzolamide preparation of preparation and gel rubber material combination, can solve its stability problem, even it becomes stable solution and does not need to make lyophilized formulations, or can reduce the consumption of stabilizing agent in each preparation, or has the phase conversion characteristic of situ-gel.

Summary of the invention

Technical problem solved by the invention is to provide several brinzolamide ophthalmic preparations, and purpose is or extends its eye holdup time, or improves its bioavailability, for reducing intraocular pressure or the treatment glaucoma provides new dosage form selection.

One, one of brinzolamide ophthalmic preparation provided of the present invention is the ocular in-situ gel agent.

The ocular in-situ gel agent comprises responsive to temperature ocular in-situ gel agent (being called for short the temperature sensing in situ gel rubber agent), the responsive ocular in-situ gel agent of pH and ion-sensitive ocular in-situ gel agent three types.

1, temperature sensing in situ gel rubber agent (being called for short preparation 1)

The disclosed prescription of temperature-sensitive situ-gel is more, mostly comparatively similar, directly loads brinzolamide with disclosed prescription and is difficult to molding or reaches prescription, also is difficult to reach the intraocular pressure lowering effect of enhancing simultaneously.

CN2005100434124 discloses prescription and the method for making of aqueous drug composition having property of reversible thermosetting gelation, and what it is characterized in that temperature sensitive substrate employing is that certain density methylcellulose and multivalence hydrochlorate form.Thermosensitive hydrogel material of the present invention is mainly selected one or more combination in any of poloxamer, chitosan, and material has had commercial product to obtain, and is convenient to industrialization; Use the brinzolamide thermosensitive hydrogel of the formula preparation of CN2005100434124, its gel phase changes do not have the present invention to announce highly sensitive simultaneously.

CN2010101485162 has announced a kind of Synergistic treatment type multi-material sustained-release eye drop and preparation method, it is characterized in that gel-type vehicle is comprised of chitosan and hyaluronic acid, and the preparation process more complicated need to can molding under the high direct voltage electrostatic interaction.By contrast, preparation technology is simpler in the present invention, is easy to industrialization.

CN2010100011309 discloses prescription and the method for making of thermo-sensitive in-situ gel pharmaceutical, and what its gel-type vehicle adopted is also chitosan, but its auxiliary gel-type vehicle use is the glucose phosphate ester.With the brinzolamide thermosensitive hydrogel of this formula preparation, its gel phase changes do not have the present invention to announce highly sensitive.

Embodiment 1 in disclosed Penciclovir ophthalmic temperature sensitivity in situ gel preparation of employing CN200710000552 and preparation method thereof prepares the temperature sensing in situ gel rubber of brinzolamide, phase transition temperature is 25 ℃, just formed gel in storage process, can't as eye drop, use, can not form situ-gel truly.The temperature sensing in situ gel rubber for preparing brinzolamide with the disclosed eye methazolamide temperature-sensitive situ-gel preparation of CN200910029314 and preparation method thereof embodiment 3, also just changed gel into mutually in room temperature, can't form situ-gel.

Thermosensitive hydrogel material and bioadhesive thickening agent polymer matrix compound use form dual system, regulate its phase transition temperature between 28～34 ℃, have higher bioavailability and lower zest.

In the 100ml gel, the component that contains following weight:

Brinzolamide or brinzolamide salt are generally 0.1～10.0g; Preferred 0.1～5.0g; 0.5～2.0g most preferably;

Gel-type vehicle is generally 0.1～40g; Preferred 5.0～30g; 10～30g most preferably;

Stabilizing agent is generally 0.1～20g; Preferred 0.1～15g; 0.1～12g most preferably;

Osmotic pressure regulator is generally 0.01～8.0g; Preferred 0.1～6.0g; 0.1～4.0g most preferably;

PH adjusting agent is generally 0.01～2.0g; Preferred 0.1～2.0g; 0.1～1.0g most preferably;

And water is supplied 100ml.

In said components, the consumption of brinzolamide or brinzolamide salt, gel-type vehicle, stabilizing agent, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described thermosensitive hydrogel substrate of preparation 1 is gel rubber material and thickening agent, and thermosensitive hydrogel substrate is selected from following material, comprises one or more the combination in any in following material: poloxamer, chitosan.

Owing to being ophthalmic preparation, for meeting the medicine preparation standard, the water in the present invention's prescription all adopts distilled water or water for injection.

For the holdup time of prolong drug at anterior corneal surface, the rate of release of regulating drug, improve the rheological property of gel rubber system, added the thickening agent polymer matrix that is no more than 5% in gel, in this scope, osmotic pressure and the viscosity of preparation meet the requirements, the adjuvant concentration increased not only can further not improved the character of preparation, cause on the contrary patient's discomfort, the described thickening agent of preparation 1 is selected from following material, comprise one or more the combination in any in following material: as dextrin and derivant, sodium alginate and derivant thereof, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose (hypromellose), hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, tyloxapol.

Described osmotic pressure regulator is at least one in mannitol, sorbitol, sodium chloride, Polyethylene Glycol, glycerol, propylene glycol or glucose.

The preparation method of responsive to temperature ocular in-situ gel agent comprises the steps:

A, get the thickening agent polymer matrix, be dissolved in water;

B, take in proportion thermosensitive hydrogel substrate again, be dissolved in water, cold preservation is placed to whole dissolvings; (illustrate: the common method that step B cold preservation used chemistry owned by France, formulation art are dissolved polymer matrix.）

C, brinzolamide or brinzolamide salt, osmotic pressure regulator and stabilizing agent are dissolved in water, filtrate and steps A and step B gained solution are merged, adjust pH to 5.0～9.0, add water to full dose, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention to stimulate eye again), single dose packing and get final product.

Wherein step C adjust pH adopts in sodium hydroxide, hydrochloric acid, boric acid, Borax, carbonate, phosphate, acetate, triethylamine any one or its mixture.

As one of preferred embodiment of the invention, responsive to temperature ocular in-situ gel of the present invention agent is, in the 100ml gel, and the component that contains following weight:

Brinzolamide or brinzolamide salt	0.5～2.0g
		Gel rubber material	10～28g
Thickening agent	0.1～2g
		Osmotic pressure regulator	0.1～4.0g
PH adjusting agent	0.1～1.0g
		Water	Be supplemented to 100ml

Wherein gel rubber material is selected from one or more combination in any of poloxamer, chitosan.The molecular weight ranges of preferred poloxamer is 5000～20000, and its oxygen ethyl and oxygen propyl group proportion are 4:1～1:4, most preferably adopt poloxamer188 (F127) and PLURONICS F87 (F68).Preferred chitosan, molecular weight is between 40,000 to 250,000, it is characterized in that deacetylation is greater than 85%, most preferably adopts deacetylation to be greater than 90%.Thickening agent is preferably from chitosan, dextrin and derivant, carbopol, hydroxypropyl emthylcellulose, hyaluronic acid, sodium alginate, tragcanth; Stabilizing agent is preferably from Tween 80, lecithin, polyvinyl alcohol, polyvinylpyrrolidone, hypromellose and edetic acid, edetate; Osmotic pressure regulator is preferably from sodium chloride, potassium chloride, glycerol, Pyrusussuriensis alcohol and glucose; PH adjusting agent is preferably from sodium dihydrogen phosphate, sodium hydrogen phosphate, boric acid, Borax, sodium carbonate, sodium bicarbonate, acetic acid, sodium acetate, sodium hydroxide, hydrochloric acid and triethylamine.

Beneficial effect: the present invention utilizes the temperature-sensitive character of poloxamer and the character of viscosity high molecular polymer, has prepared and has had suitable phase transition temperature and change mutually sensitive brinzolamide ocular in-situ gel preparation., said preparation is liquid condition at ambient temperature, with the form of solution, facilitates dropleting medicine-feeding, is easy to accurately control dosage; Temperature is elevated to while approaching body temperature and changes gel in anterior corneal surface, thereby increases medicine in the holdup time of eye, delays to eliminate, and improves the eye bioavailability; Reduce administration number of times, facilitate patient's medication.Simultaneously, this in-situ gel preparation has good biocompatibility, non-stimulated to eye.

2, the responsive ocular in-situ gel agent of pH (being called for short preparation 2)

The disclosed prescription of pH sensitive in-situ gel is more, mostly comparatively similar, directly loads brinzolamide with disclosed prescription and is difficult to molding or reaches prescription.As CN200810116084 discloses Aciclovir eye pH-sensitive in-situ gel and preparation method, what in the said preparation prescription, gel-type vehicle was selected is carbopol, used carbopol and cellulose derivative compatibility in embodiment as gel-type vehicle, but this invention is not screened to carbopol and cellulose.The present invention carries out a large amount of screenings of optimizing to carbopol and cellulosic consumption, and while using carbopol, its consumption is strict controlled in below 0.3%, just can guarantee that the product final pH can be controlled at 5～9; The embodiment 4～6 prepared with the consumption preferably and the responsive gel of the pH of embodiment A 13～A20 all have the responsive phase-change characteristic of pH preferably, and viscosity is moderate, is convenient to accurately control dosage, facilitates the patient to use.The prescription that in employing CN200810116084, embodiment 1 and embodiment 5 announce prepares respectively the pH sensitive in-situ gel of brinzolamide, and the situ-gel viscosity made is all too large, during the drop eye drip, is difficult to drip, and can't accurately control dosage.

In the 100ml gel, the component that contains following weight:

Gel-type vehicle is generally 0.1～10.0g; Preferred 0.1～8.0g; 0.1～6.0g most preferably;

And water is supplied 100ml.

The responsive gel-type vehicle of the described pH of preparation 2 comprises gel rubber material and thickening agent, wherein gel-type vehicle is selected from following material, comprises one or more the combination in any in following material: carbopol, Polycarbophil, methylcellulose, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, poloxamer, polyvinyl alcohol, hyaluronate sodium, chitosan, sodium glycerophosphate, tyloxapol.

The described thickening agent of preparation 2 is selected from following material, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol.

The mixture of gel-type vehicle preferred cellulose derivant and carbopol, weight ratio is:

Cellulose derivative: carbopol=1:0.02～1;

Preferably: cellulose derivative: carbopol=1:0.04～0.8.

Cellulose derivative comprises a kind of or its combination in any in methylcellulose, hydroxy methocel, Carboxymethyl cellulose sodium, hydroxypropyl methylcellulose, carboxymethyl cellulose, preferably hypromellose (is called for short HPMC, lower same), preferably adopt the hypromellose that viscosity is 1000～30000cP, as to block happy Kanggong department specification be the products such as HPMC E4M or HPMC EIOM.Carbopol comprises a kind of in polyacrylic acid, polyacrylate or CP, preferred CP, commodity are called carbopol, more preferably adopt the carbopol that viscosity is 2000-30000cP, as the Lubrizol company standard is the products such as carbopol 971, carbopol 980 or carbopol 981.Cellulose derivative and carbopol mix as gel-type vehicle, in aqueous solution, can form the polymer interpenetration network gel, produce significant rheology synergism, realize the slow releasing function of brinzolamide.The rheology synergistic result of hypromellose and carbopol is in Table 1.Use separately hypromellose not there is the phase conversion characteristic and use separately carbopol can not reach suitable viscosity, therefore the two mixes by certain proportion, there is good pH sensitivity and suitable viscosity, but when hypromellose concentration is excessive, the viscosity of preparation under non-physiological condition is just very large, be unfavorable for that medicine is uniformly dispersed in preparation, and be difficult to drip, can not accurately control each dose.

The rheology synergism of table 1 hypromellose and carbopol (CP980)

Sample	0.5%HPMC?E4M	0.5%HPMC?ElOM	1.0%HPMC?E4M	0.1%CP980
					Viscosity (cp, 25 ℃)	45	102	300	2440
＋0.1%CP980	3975	5355	＞9999	?

The preferred nonionic compound of described osmotic pressure regulator, as mannitol, sorbitol, glucose, glycerol or propylene glycol etc.

PH adjusting agent addition in the responsive ocular in-situ gel agent of pH of the present invention is according to the pH value to 5.0 of regulating final gel～9.0.

In said components, the consumption of brinzolamide or brinzolamide salt, gel-type vehicle, stabilizing agent, osmotic pressure regulator can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

Concrete, gel-type vehicle preferably adopts the mixture of cellulose derivative, carbopol, and weight ratio is:

Cellulose derivative: carbopol=1:0.02～1.

Preferably: cellulose derivative: carbopol=1:0.04～0.67.

In above-mentioned prescription, in order to improve the stability of preparation, gel of the present invention also can add chelating agent, as edetic acid or disodium edetate.Addition is, every 100ml gel is containing 0.01～5.0g chelating agent, and preferably addition is 0.02～1.0g.The preparation method of the responsive ocular in-situ gel agent of pH of the present invention preferably includes following steps:

A, gel-type vehicle are dissolved in the water of gross weight 25～35% and obtain the gel-type vehicle aqueous solution;

B, by brinzolamide or brinzolamide salt, osmotic pressure regulator and stabilizing agent, add the water of gross weight 25～35%, dissolve;

C, blend step A gained gel-type vehicle aqueous solution and step B gained, adjust pH to 5.0～9.0;

D, add excess water and mix, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step for except degerming, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain.

If the interpolation chelating agent adds in step B, with osmotic pressure regulator, add in the lump dissolving to get final product.

The responsive ocular in-situ gel agent of pH selects to produce the synergistic hypromellose of rheology and carbopol as mixed gel substrate, utilize two kinds of macromolecular materials to form the polymer interpenetration network gel, prepare the brinzolamide gel for eye, carbopol is low viscous liquid solution when low pH value, and contact with tear when it, the buffer capacity of tear increases its pH value, and its folding side chain opens, and viscosity increases.Because both have significant rheology synergism, lower consumption can produce full-bodied gel, thereby prolong drug is in the holdup time of eye, on the other hand, positively charged medicine brinzolamide can form complex by electrostatic interaction with electronegative carbopol, thereby further delay the release of brinzolamide from gel, produce better slow releasing function, and reduce side effects of pharmaceutical drugs.Simultaneously, gel for eye of the present invention is containing antiseptic, thus while having reduced life-time service to the zest of eye, improved drug safety.

3, ion-sensitive ocular in-situ gel agent (being called for short preparation 3)

The disclosed prescription of ion-activited in situ gel is more, mostly comparatively similar, directly loads brinzolamide with disclosed prescription and is difficult to molding or reaches prescription.As Chinese invention patent 201210203569 discloses betaxolol hydrochloride ion-sensitive type ocular in-situ gel and preparation method thereof, in the said preparation prescription, need to use antiseptic; Preparation prescription provided by the invention can not used antiseptic, the Corneal inflammation reaction of having avoided antiseptic to cause.

In the 100ml gel, the component that contains following weight:

Gel-type vehicle is generally 0.1～10.0g; Preferred 0.1～6.0g; 0.1～2.0g most preferably;

And water is supplied 100ml.

The pH value of this situ-gel compositions is 4.5 to 9.0.

The described ion-sensitive gel-type vehicle of preparation 3 comprises gel rubber material and thickening agent, and wherein gel-type vehicle can only adopt the medical hydrophilic high molecular materials such as alginate, gellan gum, deacetylated gellan gum, hypromellose to apply as substrate; Also can be using above-mentioned medical hydrophilic high molecular material and thickening agent with addition of applying as substrate.

Wherein, should adopt gellan gum in gel-type vehicle.The complexing of metal ion such as the calcium ion in itself and tear, sodium ion, potassium ion form stable hydrogen bond, and are gel state.Also can Yin Wendu and pH value change and gelation facilitates fill.

Gel-type vehicle also should adopt sodium alginate.Because gelatine can occur for alginate aqueous solution and certain density bivalence or trivalent metal ion, form the semi-solid gel.Simultaneously, sodium alginate toxicity is very low, and it is very safe being applied to the vitals such as eyes.

Thickening agent in the described gel-type vehicle of preparation 3 is selected from following polymer matrix, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol etc.

The preferred nonionic compound of osmotic pressure regulator, as mannitol, sorbitol, glucose, glycerol or propylene glycol etc.

The invention provides the preparation method of a kind of ion-sensitive ocular in-situ gel agent, comprise the steps:

By every 100ml gel containing brinzolamide or brinzolamide salt 0.1～10.0g, gel-type vehicle 0.1～10.0g, osmotic pressure regulator 0.1～4.0g, pH adjusting agent 0.1～1.0g, surplus is the water preparation.

B, by brinzolamide or brinzolamide, osmotic pressure regulator, add the water of gross weight 25～35%, dissolve;

C, blend step A gained gel-type vehicle aqueous solution and step B gained, adjust pH to 4.5～9.0;

The invention provides the preparation method of a kind of ion-sensitive ocular in-situ gel agent, preferably include following steps:

Contain brinzolamide 0.1～10.0g, gel-type vehicle 0.1～10.0g, osmotic pressure regulator 0.1～4.0g, chelating agent 0.01～0.05g by every 100g gel, pH adjusting agent 0.1～1.0g, surplus is the water preparation.

B, by brinzolamide or brinzolamide salt, chelating agent, osmotic pressure regulator, add the water of gross weight 25～35%, dissolve;

The final pH value that obtains gel is 4.5 to 9.0.

Use the prepared ion-sensitive type situ-gel of gellan gum isogel substrate, good ion-sensitive is arranged, and supplementary product consumption is few, non-physiological condition current downflow is good, is easy to splash into eye, can continue release in vitro; Alginate also is usually used in preparing the ion-sensitive type gel, has toxic and side effects little, and to advantages such as ion-sensitives, therefore, the present invention uses such material to prepare the ion-sensitive gel, the transformation sensitivity mutually of preparation, and the adjuvant toxic and side effects is little.

The ocular in-situ gel preparation that the purpose of this invention is to provide a kind of brinzolamide, the preferred responsive to temperature ocular in-situ gel agent of described preparation or the multiple gel that contains the responsive to temperature characteristic, reason is: with brinzolamide pH sensitive in-situ gel, brinzolamide ion-activited in situ gel or not containing the multiple gel phase ratio of responsive to temperature characteristic, change mutually that sensitivity is high, stability of solution good, and there is better intraocular pressure lowering effect, also less to Ocular irritation, be a kind of good glaucoma treatment preparation.

The multiple gel that contains the responsive to temperature characteristic of the present invention, comprise that temperature-pH mixes eye for the multiple in-situ gel three types by multiple in-situ gel and temperature-pH-Ar ion mixing sensitivity of responsive eye by multiple in-situ gel, temperature-Ar ion mixing sensitivity.Characteristics are: this situ-gel can splash into eyes on form as eye drop, the phase transformation that is subject to collaborative the bringing out of two or three factor in temperature, pH of tears or tear intermediate ion and produces sol-gel in conjunctival sac, while making to splash into, rarer liquid rotating becomes thicker gel.Owing to being that multiple factors brings out, can improving and change mutually sensitivity.

It is characterized in that in the 100ml gel component that contains following weight:

Brinzolamide or its pharmaceutically acceptable salt are generally 0.05～10.0g; Preferred 0.05～5.0g; 0.1～2.0g most preferably;

Gel-type vehicle:

Responsive to temperature type is generally 0.1～40g; Preferred 1.0～30g; 1.0～28g most preferably;

Or the pH responsive type is generally 0.01～10.0g; Preferred 0.1～8.0g; 0.1～6.0g most preferably;

Or ion-sensitive type is generally 0.1～10.0g; Preferred 0.1～5.0g; 0.5～2.0g most preferably;

Or other pharmaceutically acceptable eyes use composition as antiseptic, complexing of metal ion agent etc.;

And water is supplied 100ml.

The present invention also provides a kind of above-mentioned multiple gel method for preparing, the gel formula of transformation mutually by two or more combines, be worth having the gel preparation of dual or multiple sensitivity, have and better change mutually sensitivity, and increase the stability of medicine in preparation, it is characterized in that: A, gel-type vehicle are dissolved in the water of gross weight 25～35% and obtain the gel-type vehicle aqueous solution;

B, by brinzolamide or brinzolamide salt, osmotic pressure regulator, add the water of gross weight 25～35%, dissolve;

Two, one of brinzolamide ophthalmic preparation provided of the present invention is brinzolamide clathrate and pluralgel agent thereof

4, brinzolamide clathrate (being called for short preparation 4)

In 100ml clathrate preparation, the component that contains following weight:

The clathrate material is generally 1.0～50.0g; Preferred 1.0～40.0g; 1.0～30.0g most preferably;

Stabilizing agent is generally 0.1～20g; Preferred 0.1～15g; 0.1～10g most preferably;

Freeze drying protectant is generally 1.0～20.0g; Preferred 1.0～10.0g; 2.0～10.0g most preferably;

And water is supplied 100ml.

In said components, the consumption of brinzolamide, clathrate material, stabilizing agent, freeze drying protectant, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described clathrate material of preparation 4 is selected from following material, comprises one or more the combination in any in following material: at least one in cyclodextrin (CD), beta-schardinger dextrin-, HP-β-CD; Preferred beta-schardinger dextrin-and HP-β-CD; More preferably HP-β-CD.

The clathrate that uses the cyclodextrin of different materials and ratio to prepare, can solve the brinzolamide dissolubility preferably, and cyclodextrin has cornea urge saturating property, makes medicine better see through cornea barrier arrival target site.

The preparation method of brinzolamide clathrate, the optional commonly used method for preparing clathrate, as polishing, saturated solution method, freeze-drying, neutralisation, supercritical ultrasonics technology, coprecipitation, solvent evaporated method etc.The preferred solvent evaporation specifically comprises the following steps:

(1) by brinzolamide or brinzolamide salt and enclose material dissolves in organic solvent, obtain organic facies disperse medium A;

(2) stabilizing agent is scattered in the water for injection of recipe quantity, obtains aqueous phase B;

(3) eliminate in A after organic solvent by vacuum rotary steam or other modes, add aqueous phase B to dissolve, obtain formulation C;

(4) supplement distilled water to recipe quantity; add freeze drying protectant; regulate pH value and osmotic pressure; degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization; this step is degermed for removing; in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), lyophilization, obtain.

In preparation, organic solvent used is selected from following organic reagent commonly used, comprises one or more the combination in any in following material: chloroform, ethanol, dichloromethane, ethyl acetate, normal hexane, n-butyl alcohol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerin ester, acetone, acetonitrile etc.Be preferably chloroform, dichloromethane, ethyl acetate, acetone and ethanol.

Medicine is embedded in to the cavity of cyclodextrin, cyclodextrin carries medicine and is scattered in solvent, the solubilization of realization to medicine, and in certain scope, the ratio that increases cyclodextrin is conducive to improve the drug accumulation corneal permeability, clathrate of the present invention has good cornea permeability, and the enclose material is biodegradable, and toxic and side effects is little.

5, brinzolamide clathrate pluralgel agent (being called for short preparation 5)

In 100ml clathrate compound formulation, the component that contains following weight:

Gel-type vehicle is generally 0.1～40g; Preferred 0.1～30g; 0.1～28g most preferably;

And water is supplied 100ml.

In said components, the consumption of brinzolamide, clathrate material, stabilizing agent, gel-type vehicle, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described clathrate material of preparation 5 is selected from following material, comprises one or more the combination in any in following material: at least one in cyclodextrin (CD), beta-schardinger dextrin-, HP-β-CD; Preferred beta-schardinger dextrin-and HP-β-CD; More preferably HP-β-CD.

The described gel-type vehicle of preparation 5 comprises preparation 1 to 3 described gel rubber material and thickening agent, and can realize by the prescription proportioning of preparation 1 to 3 the phase conversion characteristic of situ-gel, wherein gel-type vehicle is selected from following material, comprise one or more the combination in any in following material: poloxamer, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, hyaluronate sodium, chitosan, sodium glycerophosphate, alginate, gellan gum, deacetylated gellan gum, carbopol, Polycarbophil, tyloxapol etc.Thickening agent in the described gel-type vehicle of preparation 5 is selected from following polymer matrix, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol etc.

After brinzolamide is made to clathrate, because its solution state can only be stablized the regular hour, often need further lyophilizing to make lyophilized formulations, so the present invention further combines it after considering clathrate is made to intermediate with gel, optional gel rubber material scope and preparation prescription are enumerated in preparation 1 to 3, particularly, optional way is as follows:

The preparation method of brinzolamide clathrate pluralgel agent comprises the following steps:

(4) use the water dissolution gel-type vehicle of surplus, after dissolve complete, by C and gel-type vehicle aqueous solution, regulate pH value and osmotic pressure, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add antiseptic in ophthalmic preparation of the present invention to stimulate eye again), obtain.

The brinzolamide clathrate also can adopt the described method commonly used preparation of preparation 4, as polishing, saturated solution method, freeze-drying, neutralisation, supercritical ultrasonics technology, coprecipitation etc., after said method makes the brinzolamide inclusion complex in solution, use the water for injection of surplus to dissolve gel-type vehicle, and after the two mix homogeneously is regulated to pH value and osmotic pressure, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain brinzolamide clathrate plural gel preparation.

After preparation becomes the agent of clathrate pluralgel, the stability problem that had both solved the brinzolamide inclusion complex in solution has the advantage of gel concurrently simultaneously, longer in the eye holdup time, and steadily release again, has slow releasing function preferably.

Three, one of brinzolamide ophthalmic preparation provided of the present invention is brinzolamide liposome and pluralgel agent thereof

6, brinzolamide liposome (being called for short preparation 6)

In the 100ml Liposomal formulation, the component that contains following weight:

The liposome material is generally 0.1～30.0g; Preferred 0.5～30.0g; 0.5～20.0g most preferably;

Stabilizing agent is generally 0.10～30.0g; Preferred 0.1～20.0g; 0.1～15g most preferably;

And water is supplied 100ml.

In said components, the consumption of brinzolamide or brinzolamide salt, liposome material, stabilizing agent, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described liposome material of preparation 6 is selected from following material, comprises one or more the combination in any in following material: lecithin, PHOSPHATIDYL ETHANOLAMINE (PE), fabaceous lecithin, cholesterol, cephalin, cholesterol acetyl fat, cupreol, Yolk lecithin, synthetic two palmityls-DL-α phosphatidylcholine (DPPC), synthetic phosphatidyl serine (PS), phosphatidylinositols (PI), sphingomyelin (SPH), sphingo (DCP), two myristoyl lecithin (DMPC), stearmide (SA).

The preparation method of brinzolamide Liposomal formulation selects film water legal, and concrete steps are:

(1) take liposome material and brinzolamide or the brinzolamide salt of recipe quantity, organic solvent dissolution, mix homogeneously obtains lipid soln A;

(2) organic solvent in lipid soln A is removed, made immobilized artificial membrane;

(3) stabilizing agent is scattered in the water for injection of recipe quantity, and adds in immobilized artificial membrane, aquation is complete, and high pressure homogenize or ultrasonic Treatment are adjusted the liposome particle diameter;

(4) mend and add to the full amount of water for injection, regulate pH value and osmotic pressure, (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing in degerming, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain;

(5) aseptic subpackaged aseptic liposome solutions; Or add freeze drying protectant to prepare the brinzolamide lipidosome freeze-dried preparation.

7, brinzolamide liposome pluralgel agent (being called for short preparation 7)

In 100ml liposome compound formulation, the component that contains following weight:

And water is supplied 100ml.

In said components, the consumption of brinzolamide, liposome material, stabilizing agent, gel-type vehicle, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described liposome material of preparation 7 is selected from following material, comprises one or more the combination in any in following material: lecithin, PHOSPHATIDYL ETHANOLAMINE (PE), fabaceous lecithin, cholesterol, cephalin, cholesterol acetyl fat, cupreol, Yolk lecithin, synthetic two palmityls-DL-α phosphatidylcholine (DPPC), synthetic phosphatidyl serine (PS), phosphatidylinositols (PI), sphingomyelin (SPH), sphingo (DCP), two myristoyl lecithin (DMPC), stearmide (SA).

The described gel-type vehicle of preparation 7 comprises preparation 1 to 3 described gel rubber material and thickening agent, and can realize by the prescription proportioning of preparation 1 to 3 the phase conversion characteristic of situ-gel, wherein gel-type vehicle is selected from following material, comprise one or more the combination in any in following material: poloxamer, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, hyaluronate sodium, chitosan, sodium glycerophosphate, alginate, gellan gum, deacetylated gellan gum, carbopol, Polycarbophil, tyloxapol etc.Thickening agent in the described gel-type vehicle of preparation 7 is selected from following polymer matrix, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol etc.

After brinzolamide is made to liposome, because of the time that its solution state is stable short, often need further lyophilizing to make lyophilized formulations, so the present invention further combines it after considering clathrate is made to intermediate with gel, optional material ranges and preparation prescription are enumerated in preparation 1 to 3, particularly, optional way is as follows:

The preparation method of brinzolamide Liposomal formulation comprises following methods:

(3) stabilizing agent is scattered in the water for injection of recipe quantity, and adds in immobilized artificial membrane, aquation is complete, and high pressure homogenize or ultrasonic Treatment are adjusted the liposome particle diameter, make liposome solutions B;

(4) use the water for injection of surplus to dissolve gel-type vehicle, add solution B, stir, regulate pH value and osmotic pressure, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add antiseptic in ophthalmic preparation of the present invention to stimulate eye again), obtain.

After preparation becomes the agent of liposome pluralgel, the stability problem that had both solved the brinzolamide liposome solutions has the advantage of gel concurrently simultaneously, and corneal permeability is higher, and again can steady long-acting release.

Four, one of brinzolamide ophthalmic preparation provided of the present invention is brinzolamide nanoparticle and pluralgel agent thereof

8, brinzolamide nanoparticle (being called for short preparation 8)

Although the existing lot of documents report of nano particle preparations, there is no the listing product as ophthalmic preparation.The inventor, by adopting specific nano carrier material, when improving the brinzolamide dissolubility, has improved the intraocular pressure lowering therapeutic effect greatly, obtains gratifying technique effect.

CN200610130645 discloses a kind of and has used nano eyedrop, use polylactic acid as the chitosan of material, cholesterol modification as emulsifying agent, particle diameter 283.6nm, and the release of 70 days ability is complete.WO2009073216 discloses the preparation method of brinzolamide nanoparticle, stable not, need to make freeze-dried powder.Release of the present invention is very fast, can enter efficiently eye and reach good antihypertensive effect, and not need to make lyophilized formulations and also can store for a long time.

The eye that the purpose of this invention is to provide a kind of brinzolamide is used nano particle preparations, but this nanoparticle efficient packet is carried brinzolamide, after eye drip, can in conjunctival sac, accumulate and sustained release; Can reduce the contact angle of drop and cornea, increase spreading coefficient, make preparation there is good wettability, thereby extend the time of contact of drug-carrying nanometer particle and corneal epithelium; This nanoparticle also can be pleasing to the eye by endocytosis, maybe can extend in the holdup time of eye by bio-adhesive effect or electrostatic adsorption, can effectively promote drug absorption, improves drug bioavailability, strengthens drug effect.

The eye of brinzolamide provided by the invention uses nano particle preparations, stability to be better than the nanometer formulation of literature research report, and therefore not needing to introduce additional caffolding agent makes lyophilized formulations.This nanoparticle only pharmaceutical solutions can be stored for a long time, and eye is used without special device, cost-saving, easy to use.This nanoparticle also can add the lyophilizing caffolding agent to make lyophilized formulations.

The characteristics of nanoparticle ophthalmic preparation of the present invention are with becoming to be grouped into by brinzolamide, nano carrier material, osmotic pressure regulator and pH adjusting agent or other pharmaceutically acceptable eyes, the nanoparticle envelop rate made after optimizing is up to more than 85%, particle diameter, below 400nm, has higher bioavailability, lower zest and intraocular pressure lowering effect preferably.

The purpose of this invention is to provide a kind of and use nano particle preparations, it is characterized in that in the 100ml nano particle preparations component that contains following weight:

The nanoparticle material is generally 0.1～30.0g; Preferred 0.5～30.0g; 0.5～20.0g most preferably;

Stabilizing agent is generally 0.1～30.0g; Preferred 0.1～20.0g; 0.1～15g most preferably;

Can also add other pharmaceutically acceptable eyes to use composition as antiseptic, complexing of metal ion agent etc.;

And water is supplied 100ml.

In said components, the consumption of brinzolamide, nanoparticle material, stabilizing agent, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described nanoparticle material of preparation 8 is selected from following material, comprises one or more the combination in any in following material: albumin, gelatin, chitosan, PACA and derivant thereof, PMMA and derivant thereof, PLGA and derivant thereof, PLA and derivant thereof, PCL and derivant thereof.

The described stabilizing agent of preparation 8 is selected from the combination in any of following one or more: polysorbas20, polysorbate60, Tween 80, span 20, sorbester p17, Myrij 35, sodium laurylsulfate, sodium lauryl sulphate, lecithin, polyvinyl alcohol, polyvinylpyrrolidone, crospolyvinylpyrrolidone, vinyl pyrrolidone-vinyl acetate copolymer, ethylene-vinyl acetate copolymer, poloxamer, watermiscible vitamin E (TPGS), Polyethylene Glycol, methylcellulose, ethyl cellulose, hydroxypropyl cellulose, hypromellose, sodium carboxymethyl cellulose, carbopol, sodium sulfite, sodium sulfite, sodium thiosulfate, edetic acid, edetate, arabo-ascorbic acid and sodium salt thereof, thiourea, crylic acid resin, cellulose and derivant thereof, polyoxyethylene castor oil, tyloxapol, sodium alginate, sodium tripolyphosphate, etc..Be preferably polyvinyl alcohol, TPGS, poloxamer and Tween 80; Reason is: these stabilizing agent dosage less, and prepared nanoparticle particle diameter is more even, and stability is better.

In preparation 8, organic solvent used is selected from following organic reagent commonly used, comprises one or more the combination in any in following material: chloroform, ethanol, dichloromethane, ethyl acetate, normal hexane, n-butyl alcohol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerin ester, acetone, acetonitrile etc.Be preferably chloroform, dichloromethane, ethyl acetate, acetone and ethanol.

The preparation method of brinzolamide nano particle preparations, comprise that single emulsion process, the nanometer sedimentation method or other can make any method of brinzolamide nanoparticle.

The purpose of this invention is to provide a kind of preparation method of brinzolamide nano particle preparations, comprise the following steps:

(1) the nanoparticle material of recipe quantity and brinzolamide or brinzolamide salt are dissolved in organic solvent and make organic facies A;

(2) preparation is aqueous phase B containing the solution of stabilizing agent, stirs;

(3) organic facies A is poured in aqueous phase B, adopt the physical methods such as ultrasonic or high pressure homogenize to carry out physical dispersion, make it become uniform Emulsion, or directly form the nanometer deposit seed;

(4) adopt decompression rotation steaming method or additive method to eliminate organic solvent in Emulsion (if not with an organic solvent, can save this step), degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain brinzolamide nanoparticle solution;

Or centrifugal or filter to collect nanoparticle (5), after again disperseing with the solution containing freeze drying protectant, carry out lyophilization, obtain brinzolamide nanoparticle lyophilized formulations.

Preparation method is the preparation method of optional nanoparticle commonly used also, as natural polymer polymerization, intra-liquid desiccation method, automatic emulsified method, polymeric bladder Shu Fa etc.

As one of preferred embodiment of the invention, nanoparticle of the present invention is, in the 100ml gel, and the component that contains following weight:

Brinzolamide or brinzolamide salt	0.5～2.0g
		The nanoparticle material	0.5～20.0g
Stabilizing agent	0.1～15g
		Osmotic pressure regulator	0.1～4.0g
PH adjusting agent	0.1～1.0g
		Water	Supplement 100ml

Wherein the nanoparticulate carriers material preferably certainly: PLGA and derivant thereof, PLA and derivant thereof, chitosan and derivant thereof and PCL and derivant thereof; Stabilizing agent is preferably from polyvinyl alcohol, TPGS, poloxamer and Tween 80; Osmotic pressure regulator is selected from: sodium chloride, potassium chloride, glycerol, Pyrusussuriensis alcohol and glucose; PH adjusting agent is selected from: sodium dihydrogen phosphate, sodium hydrogen phosphate, boric acid, Borax, sodium carbonate, sodium bicarbonate, acetic acid, sodium acetate, sodium hydroxide, hydrochloric acid and triethylamine.

Use nano carrier material, the brinzolamide parcel is become to nanoparticle, make that its dissolubility in water is the highest improves approximately 140 times, and the eye gone on the market at present at most only can make dissolubility in brinzolamide water improve approximately 20 times (liposome now is to the utmost unstable) with liposome.Brinzolamide nanoparticle envelop rate is up to more than 85%, and particle diameter is below 400nm, and colloid solution can steady in a long-termly be stored, and stability is better than other nano particle preparations of Liposomal formulation and bibliographical information.The brinzolamide nanoparticle also can effectively promote drug absorption, improves drug bioavailability, strengthens drug effect.

The present invention further comprises that preparation adds other art-recognized adjuvant commonly used further to make other preparation after becoming nanoparticle.

9, brinzolamide nanoparticle pluralgel agent (being called for short preparation 9)

Chinese invention patent 201110008693 discloses a kind of preparation method of pH sensitive in situ gels research nano controlled-release collyrium, this nanoparticle preparation method is comparatively complicated, grow (6～8h) consuming time, and means that need to use this difficult industry of ultrafiltration the to amplify nanoparticle that concentrates; And the position gel pH value of making is 4.0～4.5, and meta-acid, have certain zest to eye.Nanoparticle preparation technology of the present invention is simple, and short (1 hour) consuming time, technique is controlled also can amplify production, and pH is moderate, is not to use antiseptic, and eye is not had to zest.

In 100ml nanoparticle plural gel preparation, the component that contains following weight:

And water is supplied 100ml.

In said components, the consumption of brinzolamide, nanoparticle material, stabilizing agent, gel-type vehicle, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described nanoparticle material of preparation 9 is selected from following material, comprises one or more the combination in any in following material: albumin, gelatin, chitosan, PACA and derivant thereof, PMMA and derivant thereof, PLGA and derivant thereof, PLA and derivant thereof, PCL and derivant thereof.

The described gel-type vehicle of preparation 9 comprises preparation 1 to 3 described gel rubber material and thickening agent, and can realize by the prescription proportioning of preparation 1 to 3 the phase conversion characteristic of situ-gel, wherein gel-type vehicle is selected from following material, comprise one or more the combination in any in following material: poloxamer, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, hyaluronate sodium, chitosan, sodium glycerophosphate, alginate, gellan gum, deacetylated gellan gum, carbopol, Polycarbophil, tyloxapol etc.Thickening agent in the described gel-type vehicle of preparation 9 is selected from following polymer matrix, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol etc.

The invention provides a kind of preparation method of brinzolamide nanoparticle compound formulation, comprise the following steps:

(1) the nanoparticle material of recipe quantity and brinzolamide or brinzolamide salt are dissolved in to the organic facies A made in organic solvent;

(2) the water for injection preparation of use part recipe quantity is aqueous phase B containing the solution of stabilizing agent, stirs;

(3) organic facies A is poured in aqueous phase B, adopt the physical methods such as ultrasonic or high pressure homogenize to carry out physical dispersion, make it become uniform Emulsion, or directly form nanometer deposit seed C;

(4) use the water for injection of surplus to dissolve gel-type vehicle, be uniformly dispersed, the gel-type vehicle aqueous solution D of system;

(5) adopt decompression rotary evaporation or additive method to eliminate organic solvent in C (if not with an organic solvent, can save this step), again C is mixed homogeneously with D, after regulating pH value and osmotic pressure, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add antiseptic in ophthalmic preparation of the present invention to stimulate eye again), obtain brinzolamide nanoparticle plural gel preparation.

Prepare the also preparation method of optional nanoparticle commonly used of nanoparticle method, as the natural polymer polymerization, intra-liquid desiccation method, automatic emulsified method, polymeric bladder Shu Fa etc., after said method makes brinzolamide nanoparticle solution, use the water for injection of surplus to dissolve gel-type vehicle, and after the two mix homogeneously is regulated to pH value and osmotic pressure, degerming (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain brinzolamide nanoparticle plural gel preparation.

Beneficial effect of the present invention

Use the nano materials such as PLGA, the medicine parcel is become to nanometer particle, existence that can be stable in aqueous solution, and increased its dissolubility in water, as the carrier of mucosa delivery, the nanoparticle eye drop can stick on conjunctiva and cornea, make it there is preferably drug effect and can greatly extend action time, after adding gel-type vehicle, there is the phase conversion characteristic, further reach and be stranded in anterior corneal surface, reach the purpose of slow release.

This compound formulation has fully utilized the advantage of nanoparticle and in-situ gel, further improves the bioavailability of brinzolamide, thereby strengthens the intraocular pressure lowering degree and extend the intraocular pressure lowering persistent period, greatly improves patient's compliance.

Five, one of brinzolamide ophthalmic preparation provided of the present invention is brinzolamide nano suspension and pluralgel agent thereof

10, brinzolamide nano suspension (being called for short preparation 10)

After adjuvant and brinzolamide are processed through nanorize, there is good lyotropy and stability, can effectively see through anterior corneal surface, concentrate in aqueous humor, better bring into play the intraocular pressure lowering effect.

Although the existing lot of documents report of nanometer suspension preparation used in the present invention, there is no the listing product as ophthalmic preparation.At present, the prescription and the preparation technology that disclose the brinzolamide nano suspension are only arranged in WO2010146606 and WO2009073216, in WO2010146606, used dehydrated alcohol as solvent, can cause Ocular irritation, and the highest drug level of said preparation is only 1mg/ml, much smaller than listing formulation concentrations 10mg/ml; Two kinds of organic solvents of ethyl acetate and dichloromethane have been used in the preparation technology of WO2009073216, need at first form Emulsion, then organic solvent, complicated process of preparation are removed in evaporation, and organic easily being difficult for eliminates, and may there be certain toxic and side effects in residual solvent.

CN200710135285 discloses the nano suspension method for making of itraconazole, uses albumin to make stabilizing agent, and albuminous quality is not easy to control, be difficult to realize industrialization, and the step that needs emulsifying, be not so good as this patent easy, and these unstable need of writing out a prescription are made lyophilized formulations.

Use ethyl acetate when prepared by the preparation method that CN201110059685 discloses brinzolamide spherical particle nano suspension, and particle diameter is 5.176 μ m, the present invention has obvious advantage by comparison.

The inventor, by adopting specific adjuvant to form, with an organic solvent can not make the brinzolamide nano suspension, and preparation technology is simple, quality controllable; And drug level can arbitrarily be regulated in 0.05mg/ml to 50mg/ml scope; When improving the brinzolamide dissolubility, greatly improved the intraocular pressure lowering therapeutic effect, obtain gratifying technique effect.

The eye nanometer suspension preparation that the purpose of this invention is to provide a kind of brinzolamide, this nano suspension is not use any carrier, the submicron dispersion system of colloid only formed in disperse medium by pure drug particle high degree of dispersion under a small amount of stabilizer function, particle diameter is at 10～500nm.Its specific surface area is large, and medicine is stripping rapidly, effectively solves the problem that drug solubility is low.This nanometer suspension preparation adopts wet grinding, the sedimentation method, microjet method and high pressure homogenization method etc. all can make.Preferred high pressure homogenization method, this method can make the thick suspension that contains brinzolamide and stabilizing agent at a high speed by a very narrow gap, and the huge energy of utilizing cavitation, high speed shear power and interparticle collision to produce makes drug particle be pulverized as nanoparticle; Compare additive method following advantage is arranged: operate short, favorable reproducibility comparatively simple, consuming time, can be regulated the particle diameter of product by regulating homogenization pressure and cycle-index, obtain the nano suspension of different-grain diameter, and production cost be low, the products obtained therefrom particle diameter is little and even, and physical and chemical stability is good.

Nano suspension of the present invention can be stored by solution state for a long time, also can further make lyophilized formulations and store for a long time.

The purpose of this invention is to provide a kind of nano suspension, in the 100ml nano suspension, the component that contains following weight:

Brinzolamide or brinzolamide salt consumption are 0.1～10.0g; Preferred 0.1～5.0g; 0.5～2.0g most preferably;

Stabilizing agent dosage is 0.01～30.0g; Preferred 0.01～20.0g; 0.1～15.0g most preferably;

Or freeze drying protectant is generally 1.0～20.0g; Preferred 1.0～10.0g; 2.0～10.0g most preferably;

And water is supplied 100ml.

In said components, the consumption of brinzolamide or brinzolamide salt and stabilizing agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".Osmotic pressure and pH adjusting agent, or freeze drying protectant, or other pharmaceutically acceptable eyes all adopt " consumption usually " with composition.

The described brinzolamide salt of preparation comprises the arbitrary proportion combination of salt that following brinzolamide can form or any kind salt: citrate, maleate, malate, hydrochlorate, acetate, carbonate, bicarbonate, hydrophosphate, phosphoric acid hydrogen disalt, borate etc.

What stabilizing agent rose in the present invention is form nanometer formulation and reach Stabilization, be selected from following material, comprise one or more the combination in any in following material: polysorbas20, polysorbate60, Tween 80, span 20, sorbester p17, Myrij 35, sodium laurylsulfate, sodium lauryl sulphate, lecithin, polyvinyl alcohol, polyvinylpyrrolidone, crospolyvinylpyrrolidone, vinyl pyrrolidone-vinyl acetate copolymer, ethylene-vinyl acetate copolymer, poloxamer, Polyethylene Glycol, methylcellulose, ethyl cellulose, hydroxypropyl cellulose, hypromellose, sodium carboxymethyl cellulose, carbopol, sodium sulfite, sodium sulfite, sodium thiosulfate, edetic acid, the edetate edetic acid, , arabo-ascorbic acid and sodium salt thereof, thiourea, crylic acid resin, cellulose and derivant thereof, polyoxyethylene castor oil, a kind of or any mixture in tyloxapol.Preferred poloxamer, Tween 80, PVP K30, sodium carboxymethyl cellulose, acrylic resin RL100 and ethyl cellulose.

The invention provides the preparation method of a kind of brinzolamide or brinzolamide salt nano suspension, comprise the following steps:

(1) by medicine, stabilizing agent and a certain amount of solvent;

(2) use the physical methods such as dispersing emulsification machine, high pressure homogenizer, Probe Ultrasonic Searching or ball mill to make nano suspension;

(3) add osmotic pressure regulator, pH adjusting agent;

(4) filtration sterilization or directly add antiseptic;

(5) or add other pharmaceutically acceptable eyes with compositions as the complexing of metal ion agent;

(6) fill obtains the brinzolamide nano suspension or adds freeze drying protectant to make lyophilized formulations.

As one of preferred embodiment of the invention, nano suspension of the present invention is, in the 100ml nano suspension, and the component that contains following weight:

Brinzolamide or brinzolamide salt	0.5～2.0g
		Stabilizing agent	0.1～15g
Osmotic pressure regulator	0.1～4.0g
		PH adjusting agent	0.1～1.0g
Water	Be supplemented to 100ml

Wherein stabilizing agent preferably certainly: poloxamer, Tween 80, PVP K30, sodium carboxymethyl cellulose, acrylic resin RL100 and ethyl cellulose; PH adjusting agent is selected from: sodium dihydrogen phosphate, sodium hydrogen phosphate, boric acid, Borax, sodium carbonate, sodium bicarbonate, acetic acid, sodium acetate, sodium hydroxide, hydrochloric acid and triethylamine.

Beneficial effect of the present invention

The contained supplementary product kind of this nano suspension and consumption are all less, and medicine is the solid nano granule existed with certain crystal formation or unformed state; After eye drip is used, can be in conjunctival sac and anterior corneal surface be detained the long period, simultaneously higher medicine is dense to be conducive to the cornea medicine that Concentraton gradient relies on and to see through, and improves bioavailability, extends action time, improves curative effect.Nano suspension is compared with common suspensoid, and particle diameter is less, and drug-eluting speed is faster, ocular absorption speed and degree are just larger, and the drug particles in nano suspension has larger surface area, at eye, has certain adhesion, can improve the degree of absorption of medicine, extend action time.

11, brinzolamide nano suspension pluralgel agent (being called for short preparation 11)

In 100ml nano suspension plural gel preparation, the component that contains following weight:

Or freeze drying protectant is generally 1.0～20.0g; Preferred 1.0～10.0g; 2.0～10.0g most preferably; ;

And water is supplied 100ml.

In said components, the consumption of brinzolamide, stabilizing agent, gel-type vehicle, osmotic pressure and pH adjusting agent can adopt the combination in any of " consumption usually ", " preferable amount ", " most preferred quantities ".

The described stabilizing agent of preparation 11 is selected from following material, comprise one or more the combination in any in following material: polysorbas20, polysorbate60, Tween 80, span 20, sorbester p17, Myrij 35, sodium laurylsulfate, sodium lauryl sulphate, lecithin, polyvinyl alcohol, polyvinylpyrrolidone, crospolyvinylpyrrolidone, vinyl pyrrolidone-vinyl acetate copolymer, ethylene-vinyl acetate copolymer, poloxamer, Polyethylene Glycol, methylcellulose, ethyl cellulose, hydroxypropyl cellulose, hypromellose, sodium carboxymethyl cellulose, carbopol, sodium sulfite, sodium sulfite, sodium thiosulfate, edetic acid, edetate, arabo-ascorbic acid and sodium salt thereof, thiourea, crylic acid resin, cellulose and derivant thereof, polyoxyethylene castor oil, a kind of or any mixture in tyloxapol.Stabilizing agent is called again suspending agent sometimes, and the differentiation of suspending agent and stabilizing agent mainly on its consumption, and has different being used in conjunction with, and in embodiment, will illustrate.

The described gel-type vehicle of preparation 11 comprises preparation 1 to 3 described gel rubber material and thickening agent, and can realize by the prescription proportioning of preparation 1 to 3 the phase conversion characteristic of situ-gel, wherein gel-type vehicle is selected from following material, comprise one or more the combination in any in following material: poloxamer, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, hyaluronate sodium, chitosan, sodium glycerophosphate, alginate, gellan gum, deacetylated gellan gum, carbopol, Polycarbophil, tyloxapol etc.Thickening agent in the described gel-type vehicle of preparation 11 is selected from following polymer matrix, comprises one or more the combination in any in following material: as polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropyl emthylcellulose, hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, sodium alginate and derivant thereof, arabic gum, tragcanth, guar gum, carbopol, phytohemagglutinin, hyaluronic acid, xanthan gum, chitosan, dextrin and derivant, tyloxapol etc.

The invention provides the preparation method of a kind of brinzolamide or the agent of brinzolamide salt nanometer suspension pluralgel, comprise the following steps:

(1) by medicine, stabilizing agent and a certain amount of solvent;

(2) use the physical methods such as dispersing emulsification machine, high pressure homogenizer, Probe Ultrasonic Searching or ball mill to make nano suspension A;

(3) use the water for injection of surplus to dissolve gel-type vehicle, stir, obtain gel-type vehicle aqueous solution B, A and B are mixed, regulate pH value and osmotic pressure, (can adopt the degerming modes commonly used such as 0.22 μ m filtering with microporous membrane or radiation sterilization, this step is degermed for removing in degerming, in order to avoid add again antiseptic in ophthalmic preparation of the present invention to stimulate eye), obtain.

The particle diameter of nanometer suspension pluralgel agent can be regulated with the change cycle-index by power and the ultrasonic time that changes Probe Ultrasonic Searching and the pressure of regulating high pressure homogenize.

Beneficial effect of the present invention

This compound formulation has fully utilized the advantage of nano suspension and in-situ gel, further improves the bioavailability of brinzolamide, thereby strengthens the intraocular pressure lowering degree and extend the intraocular pressure lowering persistent period, greatly improves patient's compliance.

Further illustrate:

Preparation 1 to the described brinzolamide salt of preparation 11 comprises the arbitrary proportion combination of salt or any kind salt of the feasible one-tenth of following brinzolamide salt: citrate, maleate, malate, hydrochlorate, acetate, carbonate, bicarbonate, hydrophosphate, phosphoric acid hydrogen disalt, borate etc.

Preparation 1 to the described stabilizing agent of preparation 11 is selected from following material, comprise one or more the combination in any in following material: polysorbas20, polysorbate60, Tween 80, span 20, sorbester p17, Myrij 35, sodium laurylsulfate, sodium lauryl sulphate, lecithin, polyvinyl alcohol, polyvinylpyrrolidone, crospolyvinylpyrrolidone, vinyl pyrrolidone-vinyl acetate copolymer, ethylene-vinyl acetate copolymer, poloxamer, Polyethylene Glycol, methylcellulose, ethyl cellulose, hydroxypropyl cellulose, hypromellose, sodium carboxymethyl cellulose, carbopol, sodium sulfite, sodium sulfite, sodium thiosulfate, edetic acid, edetate, arabo-ascorbic acid and sodium salt thereof, thiourea, crylic acid resin, cellulose and derivant thereof, polyoxyethylene castor oil, tyloxapol.

Preparation 1 to the described osmotic pressure regulator of preparation 11 is selected from following material, comprises one or more the combination in any in following material: sodium chloride, glucose, sodium acetate, sodium carbonate, sodium bicarbonate, sodium hydrogen phosphate, sodium dihydrogen phosphate, boric acid, Borax, propylene glycol, glycerol, mannitol.

Preparation 1 to the freeze drying protectant of preparation 11 is selected from following material, comprises one or more the combination in any in following material: polyvinylpyrrolidone, mannitol, sucrose, lactose, glucose, Polyethylene Glycol, glucosan, trehalose, albumin.

Preparation 1 to 11 also can add chelating agent, is selected from following material, comprises one or more the combination in any in following material: sodium pyrosulfite, sodium sulfite, sodium sulfite, sodium thiosulfate, edetic acid, disodium edetate, sodium citrate.

Preparation 1 to the described pH adjusting agent of preparation 11 is selected from following material, comprises one or more the combination in any in following material: hydroxide, organic amine and or mineral acid and salt, organic acid and salt thereof, sodium carbonate, sodium bicarbonate, sodium hydrogen phosphate, sodium dihydrogen phosphate, boric acid, Borax.

Preparation 1 to organic solvent used in preparation 11 is selected from following organic reagent commonly used, comprises one or more the combination in any in following material: ethanol, dichloromethane, ethyl acetate, normal hexane, n-butyl alcohol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerin ester, acetone etc.

Brinzolamide ophthalmic preparation of the present invention, can be used for the treatment of the diseases such as high intraocular pressure and glaucoma, and dosage is generally 1 times/day, and each giving suffered from 1 of eye, concrete, can, according to patient's the state of an illness, age etc., by the doctor, be determined.Irritant to eyes because of antiseptic, especially may cause serious zest during life-time service.Therefore, brinzolamide ophthalmic preparation of the present invention, containing antiseptic, is not disposable unit dose package sterile preparation.

To sum up, the present invention provides multiple slow release brinzolamide ophthalmic preparation for glaucoma and high intraocular pressure patient, and said preparation has the characteristics of long-time slowly release and foreign sense, can significantly improve patient's compliance; And more lasting than commercial preparation intraocular pressure lowering better effects if, action time; Simultaneously, preparation used does not add any antiseptic, adopts unit dose package, has reduced the zest of eye drop life-time service to eye, has greatly improved the safety of medication.This long-acting slow-release preparation technology also can be used for the medicine of other local use, realizes slow release long-acting.

The accompanying drawing explanation

Before and after Fig. 1 brinzolamide situ-gel 24% poloxamer188 and the dilution of 0.5% PLURONICS F87 compatibility tear, viscosity changes.

Fig. 2 brinzolamide pH responsive type gel A, B viscosity when different pH value changes

A, 0.1% carbopol 980-0.5%HPMC E4M-1% brinzolamide gel

B, 0.2% carbopol 980-0.5%HPMC E4M-1% brinzolamide gel

The comparison of tri-kinds of gel tablets in vitros of Fig. 3 and solution, suspensoid (thermosensitive in situ gel-embodiment 1, the responsive gel-embodiment 4 of pH, ion-sensitive gel-embodiment 8).

Tri-kinds of gels of Fig. 4 and solution isolated cornea see through accumulation medication amount result (thermosensitive in situ gel-embodiment 1, the responsive gel-embodiment 4 of pH, ion-sensitive gel-embodiment 8).

Fig. 5 variable concentrations enclose material sees through the result of accumulation medication amount to the rabbit isolated cornea.

The intraocular pressure reduction value of Fig. 6-1 temperature-sensitive situ-gel (embodiment A 6) and commercial preparation contrast (" * " means p<0.05).

Fig. 6-2 temperature-sensitive situ-gel (embodiment A 6) is with respect to intraocular pressure reduction value and the commercial preparation contrast (" * " means p<0.05) of the responsive ocular in-situ gel agent of pH (embodiment A 16) and ion-sensitive ocular in-situ gel agent (embodiment A 25).

Fig. 6-3 temperature-pH mixes intraocular pressure reduction value with multiple in-situ gel (embodiment A 38) and the commercial preparation contrast (" * " mean p<0.05) by multiple in-situ gel (embodiment A 37) and temperature-pH-Ar ion mixing sensitivity by multiple in-situ gel (embodiment A 36), pH-Ar ion mixing sensitivity of responsive eye by multiple in-situ gel (embodiment A 35), temperature-Ar ion mixing sensitivity.

The intraocular pressure reduction value percentage ratio of Fig. 7 brinzolamide nanoparticle (Embodiment B 12) and commercial preparation contrast (" * " means p<0.05).

The intraocular pressure reduction value percentage ratio of Fig. 8 nano suspension (Embodiment C 24) and commercial preparation contrast (" * " means p<0.05).

Fig. 9-1 nano suspension (Embodiment C 24) and nano suspension thermosensitive in situ gel (embodiment D19) intraocular pressure reduction value contrast (" * " means p<0.05).

Fig. 9-2 nanoparticle gel compound formulation (embodiment D6), nanometer suspension gel compound formulation (embodiment D19) and clathrate-pH responsive gel intraocular pressure reduction value contrast (" * " means p<0.05).

Fig. 9-3 nanoparticle gel compound formulation (embodiment D6), nanometer suspension gel compound formulation (embodiment D19) and commercial preparation contrast (" * " means p<0.05).

The specific embodiment

Below by specific description of embodiments of the present invention the explanation but do not limit the present invention.

Embodiment 1～3

Annotate: "/" means not add.

Embodiment 1 to 3 is brinzolamide or brinzolamide salt responsive to temperature type gel prescription, and method for making takes the thickening agent polymer matrix as follows in proportion, adds distilled water, fully stirs, and is placed to whole dissolvings; Take in proportion again poloxamer, add in distilled water, stir and make to be partly dissolved, be placed to whole dissolvings in cold compartment of refrigerator; Brinzolamide or brinzolamide and solubilizing agent, osmotic pressure regulator, antiseptic are dissolved in distilled water, stirring makes its dissolving, through 0.22 μ m filtering with microporous membrane, filtrate and above-mentioned macromolecular solution are merged, adding distil water is to full dose, the pH value of pH adjusting agent regulator solution of take is 5.0～9.0, stirs, and obtains.

Embodiment 4～6

Embodiment 4 to 6 is brinzolamide or brinzolamide salt pH responsive type gel prescription, method for making is scattered in soaked overnight in 30ml water for injection by hypromellose and carbopol as follows, stir, obtain solution, standby, by brinzolamide or brinzolamide salt, osmotic pressure regulator, inject water 30ml, stir lower the dissolving, 0.22 μ m microporous filter membrane filters, filtrate is joined in the hypromellose and carbopol solution of above-mentioned preparation, add water for injection to 100ml, the pH that adds appropriate pH adjusting agent solution to be adjusted to system is 5.5, and stir, obtain the brinzolamide gel for eye.

Embodiment 7～9

Annotate: "/" means not add.

Embodiment 7 to 9 is brinzolamide or brinzolamide salt ion responsive type gel prescription, method for making is as follows: the macromolecular material of recipe quantity is added in 20 to 30ml waters for injection, magnetic agitation, after being uniformly dispersed, by medicine, osmotic pressure regulator, inject water 30ml, stir lower the dissolving, 0.22 μ m microporous filter membrane filters, and filtrate is joined in the hypromellose and carbopol solution of above-mentioned preparation, regulate pH value and osmotic pressure, and add water for injection to 100ml, and stir, obtain the brinzolamide gel for eye.

The applicant further researchs and analyses above-mentioned three kinds of situ-gels in the phase in priority.

The agent of embodiment A 1～A10 brinzolamide responsive to temperature ocular in-situ gel

Annotate: "/" means not add.

Embodiment A 1 to A5 brinzolamide or brinzolamide salt responsive to temperature type gel prescription, method for making takes the thickening agent polymer matrix as follows in proportion, adds distilled water, fully stirs, and is placed to whole dissolvings; Take in proportion again poloxamer, add in distilled water, stir and make to be partly dissolved, be placed to whole dissolvings in cold compartment of refrigerator; Brinzolamide or brinzolamide and solubilizing agent, osmotic pressure regulator are dissolved in distilled water, stirring makes its dissolving, through 0.22 μ m filtering with microporous membrane, filtrate and above-mentioned macromolecular solution are merged, adding distil water is to full dose, the pH value of pH adjusting agent regulator solution of take is 5.0～9.0, stirs, and obtains.

Embodiment A 6 to A10 is brinzolamide or brinzolamide salt responsive to temperature type gel prescription, and method for making takes the thickening agent polymer matrix as follows in proportion, adds distilled water, fully stirs, and is placed to whole dissolvings; Take the chitosan of recipe quantity, be scattered in previously prepared 0.1M hydrochloric acid, magnetic agitation makes its dissolving in 24 hours; Brinzolamide or brinzolamide and solubilizing agent, osmotic pressure regulator are dissolved in distilled water, stirring makes its dissolving, through 0.22 μ m filtering with microporous membrane, filtrate and above-mentioned macromolecular solution are merged, adding distil water is to full dose, the pH value of pH adjusting agent regulator solution of take is 5.0～9.0, stirs, and obtains.

The responsive ocular in-situ gel agent of embodiment A 13～A20 brinzolamide pH

Annotate: "/" means not add.

Embodiment A 13 to A20 is brinzolamide or brinzolamide salt pH responsive type gel prescription, method for making is scattered in soaked overnight in 30ml water for injection by hypromellose and carbopol as follows, stir, obtain solution, standby, by brinzolamide or brinzolamide salt, osmotic pressure regulator, inject water 30ml, stir lower the dissolving, 0.22 μ m microporous filter membrane filters, filtrate is joined in the hypromellose and carbopol solution of above-mentioned preparation, adding the water for injection ad pond om is 100ml, the pH that adds appropriate pH adjusting agent solution to be adjusted to system is 5.5, and stir, obtain the brinzolamide gel for eye.

The agent of embodiment A 21～A30 brinzolamide ion-sensitive ocular in-situ gel

Annotate: "/" means not add.

Embodiment A 21 to A30 is brinzolamide or brinzolamide salt ion responsive type gel prescription, and method for making is as follows: the macromolecular material of recipe quantity is added in 20 to 30ml waters for injection to magnetic agitation, after being uniformly dispersed, by medicine, osmotic pressure regulator, inject water 30ml, stir lower the dissolving, 0.22 μ m microporous filter membrane filters, and regulates pH value and osmotic pressure, and to add the water for injection ad pond om be 100ml, stir, obtain the brinzolamide gel for eye.The system gel add artificial tears (sodium chloride: 8.3g, calcium chloride dihydrate: 0.084g, potassium chloride: 1.4g, water: 1L), all can form gel by 40:7.

The multiple gel preparation of embodiment A 31～A40

Annotate: "/" means not add; "-" means not detect.

Embodiment A 31 to A40 is the multiple responsive gel prescription of brinzolamide or brinzolamide salt, and method for making is as follows: the macromolecular material of recipe quantity is added in 20 to 30ml waters for injection to magnetic agitation, after being uniformly dispersed, by medicine, osmotic pressure regulator, inject water 30ml, stir lower the dissolving, 0.22 μ m microporous filter membrane filters, and regulates pH value and osmotic pressure, and to add the water for injection ad pond om be 100ml, stir, obtain the brinzolamide gel for eye.

Experimental example A1 pharmacodynamic evaluation

Get the normal rabbits random packet, 6 every group, male and female half and half.The bilateral eye gives 50 μ l Oxybuprocaines (times promise happiness, Japan) eye anesthesia, uses Tono-

tonometer is measured eyes baseline intraocular pressure, is designated as 0 moment intraocular pressure.One-sided eye gives 50 μ l preparations (test formulation: temperature-sensitive situ-gel (embodiment A 6), the responsive ocular in-situ gel agent (embodiment A 16) of pH, ion-sensitive ocular in-situ gel agent (embodiment A 25), temperature-pH is mixed responsive eye with multiple in-situ gel (embodiment A 35), the eye of temperature-Ar ion mixing sensitivity is with multiple in-situ gel (embodiment A 36), the eye of pH-Ar ion mixing sensitivity is with multiple in-situ gel (embodiment A 37), the group that the eye of temperature-pH-Ar ion mixing sensitivity is produced with multiple in-situ gel (embodiment A 38) or commercially available suspensoid Alcon Universal Ltd. is stood

) to experimental group rabbit eyes conjunctival sac.30,60,120,180,240 and 300 minutes minutes mensuration bilateral varieties of intraocular pressure after administration, and calculate intraocular pressure reduction value (mmHg): intraocular pressure reduction value=certain moment intraocular pressure basis intraocular pressure.Adopt the t check in SPSS13.0 software to carry out statistical procedures, p<0.05 use " * " sign.

Obtain intraocular pressure reduction value and the commercial preparation contrast (" * " mean p<0.05) of result referring to Fig. 6-1 temperature-sensitive situ-gel, conclusion is that the intraocular pressure lowering successful of thermosensitive hydrogel is better than commercial preparation, and the intraocular pressure value descends remarkable and action time is lasting; Fig. 6-2 temperature-sensitive situ-gel is with respect to intraocular pressure reduction value and the commercial preparation contrast (" * " means p<0.05) of the responsive ocular in-situ gel agent of pH and the agent of ion-sensitive ocular in-situ gel, conclusion is: the strong and weak order of the intraocular pressure lowering effect of three kinds of gels is: thermosensitive hydrogel > the responsive gel of pH > ion-sensitive gel, wherein thermosensitive hydrogel intraocular pressure value decline Chengdu and continuous action time optimal; Fig. 6-3 temperature-pH mixes intraocular pressure reduction value with multiple in-situ gel and the commercial preparation contrast (" * " mean p<0.05) by multiple in-situ gel and temperature-pH-Ar ion mixing sensitivity by multiple in-situ gel, pH-Ar ion mixing sensitivity of responsive eye by multiple in-situ gel, temperature-Ar ion mixing sensitivity, conclusion is that the intraocular pressure lowering effect of 4 kinds of gels all is better than commercial preparation, not longer containing the responsive reducing iop persistent period of the pH-Ar ion mixing of thermosensitive hydrogel containing 3 multiple gel preparation effects of thermosensitive hydrogel.

Embodiment 10～12

Embodiment 10 to 12 is preparation 4 brinzolamide clathrate prescriptions, method for making is as follows: the brinzolamide of recipe quantity and HP-β-CD are added in 20 to 30ml ethanol, magnetic agitation, after being uniformly dispersed, be placed in 25 ℃ of Rotary Evaporators and boil off organic solvent formation thin film, stabilizing agent is dissolved, add dissolving films, and to add the water for injection ad pond om be 100 grams, stir, use pH value regulator and osmotic pressure regulator, regulate pH value and osmotic pressure, 0.22 μ m microporous filter membrane filters, filtrate is added to freeze drying protectant, stir, with-80 ℃ of pre-freezes 24 hours, lyophilization, obtain brinzolamide clathrate preparation.

Embodiment 13～15

Annotate: "/" means not add.

Embodiment 13 to 15 is prescriptions of preparation 6 brinzolamide liposomees, and method for making is as follows: take phospholipid, cholesterol and the medicine of recipe quantity, and dissolve with ethanol, the mix homogeneously mixed liquor is on Rotary Evaporators, and the pressure reducing and steaming organic solvent, prepare immobilized artificial membrane; To the water for injection that is dissolved with stabilizing agent that adds 50% in immobilized artificial membrane, aquation is complete, and high pressure homogenize or ultrasonic Treatment are adjusted the liposome particle diameter; Benefit adds to the full amount of water for injection, and regulates pH value and osmotic pressure, and filtering with microporous membrane, obtain, aseptic subpackaged or add freeze drying protectant to prepare the brinzolamide lipidosome freeze-dried preparation.

Embodiment 16～18

Annotate :/mean not add.

Embodiment 16 to 18 is prescriptions of preparation 8 brinzolamide nanoparticles, preparation method is as follows: the material of recipe quantity and medicine are dissolved in to the organic facies of making in dichloromethane, the solution containing stabilizing agent of preparation is water, organic facies is poured in water, 10000rpm stirred after 3～5 minutes, in Probe Ultrasonic Searching cell pulverization machine, ultrasonic 5 minutes of 100W, obtain Emulsion, the decompression rotation is steamed and is removed organic solvent, 0.22 the degerming of μ m filtering with microporous membrane, carry out lyophilization after the sucrose solution with 2%～10% disperses again, obtains the brinzolamide nanometer formulation.

In the phase, to nano particle preparations, further research is as follows in priority for the applicant:

Embodiment B 1～B6 brinzolamide consumption is preferred

Annotate :=expression is suitable with commercially available suspensoid drug effect ,+mean that drug effect is better than the degree of commercially available suspensoid.

Brinzolamide and PCL are dissolved in the mixed solvent of dichloromethane and acetone, add in polyvinyl alcohol water solution, under condition of ice bath, the 300W Probe Ultrasonic Searching is minutes 5 minutes, obtain O/W Emulsion, 37 ℃ of water-bath distilling under reduced pressure eliminate organic solution, regulate pH value and osmotic pressure, or add antiseptic, add water and be settled to 100ml, obtain.Carry out the intraocular pressure lowering effect relatively with commercially available suspensoid, the result demonstration, when brinzolamide is 0.05mg/100ml, drug effect and commercially available suitable, improve consumption, drug effect significantly is better than commercially available, and, along with concentration increases, drug effect increases gradually, when concentration reaches 2mg/100ml, drug action reaches peak value, further increases drug level, and drug effect no longer includes significant change.

Embodiment B 7～B16 nano carrier material is preferred

Annotate: "/" means not add.

Brinzolamide and nanoparticulate carriers material are dissolved in organic solvent, add in the aqueous solution that contains stabilizing agent, 300W Probe Ultrasonic Searching 5 minutes or magnetic agitation under condition of ice bath, obtain O/W Emulsion or directly obtain nanoparticle, 37 ℃ of water-bath distilling under reduced pressure eliminate organic solution, regulate pH value and osmotic pressure, or add antiseptic, add water and be settled to 100ml, obtain.

Embodiment B 17～B22 nano carrier material is preferred

Annotate: "/" means not add.

Brinzolamide is mixed with adjuvant, add in the aqueous solution that contains stabilizing agent, high pressure homogenize or dispersant disperse, and directly obtain nanoparticle, regulate pH value and osmotic pressure, or add antiseptic, add water and are settled to 100ml, obtain.

Experimental example B1 pharmacodynamic evaluation

Test operation is with test example A1.The used test preparation is: Embodiment B 12.

Obtain intraocular pressure reduction value percentage ratio and the commercial preparation contrast (" * " mean p<0.05) of result referring to Fig. 7 brinzolamide nanoparticle (Embodiment B 12), conclusion is that the intraocular pressure lowering successful of brinzolamide nanoparticle is better than commercial preparation, and the intraocular pressure value descends significantly.

Embodiment 19～21

Annotate: "/" means not add.

Embodiment 19 to 21 is prescriptions of preparation 10 brinzolamide nano suspensions, embodiment 19 and 20 preparation method: medicine and adjuvant are dissolved in organic facies, use high-shear emulsion machine 10000rpm, emulsifying is sheared 5 minutes, obtain just suspension, high pressure homogenize 300bar, circulate 6 times, 500bar circulation 6 times, 800bar circulation 6 times, 1000bar circulation 20 times, regulate pH value and osmotic pressure and osmotic pressure, cross 0.22 μ m filter membrane, obtain.

The preparation method of embodiment 21 is: medicine and adjuvant are dissolved in organic facies, use high-shear emulsion machine 6000rpm, emulsifying is sheared 5 minutes, obtains just suspension, and under ice bath, Probe Ultrasonic Searching 90W is ultrasonic 5 minutes,, cross 0.22 μ m filter membrane, obtain.

Embodiment C 1～C6 brinzolamide consumption is preferred

Brinzolamide and PVP K30 are scattered in water, use high-shear emulsion machine to shear 5 minutes, obtain thick suspension, high pressure homogenize 500bar circulation 5 times, 1000bar circulation 5 times, regulate pH and osmotic pressure, add complexing of metal ion agent and antiseptic, add water and be settled to 100ml, obtain.Carry out the intraocular pressure lowering effect relatively with commercially available suspensoid, the result demonstration, when brinzolamide is 0.05mg/100ml, drug effect and commercially available suitable, improve consumption, drug effect significantly is better than commercially available, and, along with concentration increases, drug effect increases gradually, when concentration reaches 2mg/100ml, drug action reaches peak value, further increases drug level, and drug effect no longer includes significant change.

Embodiment C 7～C15 stabilizing agent is preferred

Annotate: "/" means not add.

Brinzolamide is mixed with adjuvant, and be added in the quartzy cup of ball mill with a certain amount of water, the use zirconium oxide is abrasive media, 35Hz grinds 6 hours, takes out preparation, removes by filter abrasive media, regulate pH value and osmotic pressure, add complexing of metal ion agent or antiseptic, add water and be settled to 100ml, obtain.The particle size determination result shows, while with glycerol, making stabilizing agent, particle diameter is too large and distributed pole is inhomogeneous, and therefore, glycerol is not selected as stabilizing agent.

Annotate: "/" means not add.

Brinzolamide is mixed with adjuvant, and be added in the quartzy cup of ball mill with a certain amount of water, the use zirconium oxide is abrasive media, 35Hz grinds 6 hours, takes out preparation, removes by filter abrasive media, regulate pH value and osmotic pressure, add complexing of metal ion agent or antiseptic, add water and be settled to 100ml, obtain; Or add freeze drying protectant, and make lyophilized formulations, obtain.The demonstration of particle size determination result, during with the single stable agent, the nano suspension particle diameter that glycerol makes is too large and distributed pole is inhomogeneous, only can stablize 1 day, and therefore, glycerol is not selected as stabilizing agent.When poloxamer, Tween 80, PVP K30, sodium carboxymethyl cellulose, acrylic resin RL100 and ethyl cellulose are alone, Jun stabilization time of nano suspension>=2 months, as preferred stabilizing agent.

Experimental example C1 pharmacodynamic evaluation

Test operation is with test example A1.The used test preparation is: Embodiment C 24.

Obtain intraocular pressure reduction value percentage ratio and the commercial preparation contrast (" * " mean p<0.05) of result referring to Fig. 8 nano suspension (Embodiment C 24)., conclusion is that the intraocular pressure lowering successful of brinzolamide nano suspension is better than commercial preparation, the intraocular pressure value descends significantly.

Embodiment 22～25

Annotate: "/" means not add.

Embodiment 22 to 25 is the agent of brinzolamide pluralgel, embodiment 22 prescriptions are selected from preparation 5, preparation method: get the brinzolamide of recipe quantity and enclose material dissolves in organic solvent, obtain organic facies disperse medium A, stabilizing agent is scattered in the water for injection of recipe quantity, obtain aqueous phase B, drip slowly organic facies disperse medium A in B, stir, mixed liquor is removed organic solvent, obtain formulation C, use the water dissolution gel-type vehicle of surplus, after dissolve complete, C and gel-type vehicle aqueous solution is even, regulate pH value and osmotic pressure, 0.22 μ m filtering with microporous membrane degerming, obtain.

Embodiment 23 and 24 is selected from preparation 11, preparation method: the adjuvants such as medicine and suspending agent are dissolved in organic solvent, obtain organic facies disperse medium A; Stabilizing agent is scattered in the water for injection of recipe quantity, obtains aqueous phase B; Drip slowly organic facies disperse medium A in aqueous phase B, use the physical method such as mulser to prepare the suspensoid colostrum simultaneously, then use the physical methods such as Probe Ultrasonic Searching or high pressure homogenize to increase the stability of suspensoid colostrums, obtain suspension emulsion C; Remove the organic solvent in C, obtain whole suspensoid D; Use the water for injection of surplus to dissolve gel-type vehicle, stir, obtain gel-type vehicle aqueous solution E, D and E are mixed, regulate pH value and osmotic pressure, 0.22 μ m filtering with microporous membrane degerming, obtain;

Embodiment D1～D10 nanometer particle in-situ gel compound formulation

Annotate: "/" means not add.

The preparation method of brinzolamide compound formulation, comprise the following steps: the standby nanoparticle of the following legal system of water that uses recipe quantity 50%～80%: brinzolamide and nanoparticulate carriers material are dissolved in organic solvent, add in the aqueous solution that contains stabilizing agent, 300W Probe Ultrasonic Searching 5 minutes or magnetic agitation under condition of ice bath, obtain O/W Emulsion or directly obtain nanoparticle, 37 ℃ of water-bath distilling under reduced pressure eliminate organic solution, then use the water of recipe quantity 20%～50% to prepare in-situ gel preparation, nanometer formulation is scattered in situ-gel, regulate pH value and osmotic pressure, or add antiseptic, obtain.

Embodiment D11～D20 nano suspension situ-gel

Annotate: "/" means not add.

The preparation method of brinzolamide compound formulation, comprise the following steps: the standby nano suspension of the following legal system of water that uses recipe quantity 50%～80%: brinzolamide is mixed with adjuvant, and be added in the quartzy cup of ball mill with a certain amount of water, the use zirconium oxide is abrasive media, 35Hz grinds 6 hours, take out preparation, remove by filter abrasive media, add gel-type vehicle and stabilizing agent, regulate pH value and osmotic pressure, add complexing of metal ion agent or antiseptic, moisturizing, to 100ml, obtains.

Experimental example D1 pharmacodynamic evaluation

Test operation is with test example A1.The used test preparation is: nanoparticle solution suspensoid (Embodiment C 24) and nanoparticle ion suspensoid-thermosensitive in situ gel (embodiment D19).

Obtain result referring to Fig. 9-1 nanoparticle solution suspensoid (Embodiment C 24) and nanoparticle ion suspensoid thermosensitive in situ gel (embodiment D19) intraocular pressure reduction value contrast (" * " means p<0.05), conclusion be nano suspension and thermosensitive in situ gel compound after, the intraocular pressure lowering effect is still fine, and can maintain the longer time; Fig. 9-2 nanoparticle gel compound formulation (embodiment D6), nanometer suspension gel compound formulation (embodiment D19) and clathrate-pH responsive gel intraocular pressure reduction value contrast (" * " means p<0.05), conclusion is: 3 kinds of compound formulations are by certain intraocular pressure lowering effect, and continuous action time length is sequentially: nanoparticle gel compound formulation > nanometer suspension gel compound formulation > the responsive gel of clathrate-pH; Fig. 9-3 nanoparticle gel compound formulation (embodiment D6), nanometer suspension gel compound formulation (embodiment D19) and commercial preparation contrast (" * " means p<0.05), conclusion is: the intraocular pressure lowering effect of nanoparticle gel compound formulation and nanometer suspension gel compound formulation all is better than commercial preparation.

The phase transition detection of experimental example 1 brinzolamide temperature, pH and ion-sensitive gel for eye use

1.1 the temperature sensitive hydrogel phase transition temperature is measured:

Gel solution is added in cillin bottle, after adding magnetic stir bar, bottle is placed in to the water bath device with magnetic agitation, controlling magnetic agitation speed is 100rpm, then regulate temperature, 0.5 ℃ of rising per minute, maintain 2 minutes, observe temperature record that stirrer stops operating, measure calculating mean value 3 times.

The brinzolamide gel preparation gel phase transition temperature measurement result (n=3) of the different prescriptions of table 1-1

Table 1-1 result shows: add hypromellose can reduce to a certain extent the gelation temperature of Poloxamer 127.With the Poloxamer 127 of alone same concentrations, compare, the viscosity of gel increases, the effect that viscosity increases is relevant to addition, prescription all can form gel in preference temperature, the material mixture ratio of change prescription, as increase the ratio of Poloxamer 127, the gel phase transition temperature reduces, and higher to the conditional request of producing, transporting and storing, cost increases, and use inconvenience, and the ratio of reduction Poloxamer 127, its phase transition temperature raises, and the temperature of eyeball surface is 34 ℃ to 37 ℃, the requirement that it changes mutually can not be met, gel can't be formed.

1.2pH the responsive type gel phase changes the mensuration of pH value:

Under 34 ± 0.2 ℃, the viscosity of gel when pH value being set respectively being 4.0-9.0 and measuring this pH value, using its viscosity and become suddenly large point as changing mutually the pH point, as shown in Figure 2.

The different prescription of table 1-2 brinzolamide gel preparation gel phase changes pH measurement result (n=3)

Table 2 result shows: the amount of carbomer increases, gel phase changes the pH value step-down, but still in suitable scope, there is gel characteristic preferably, if continue to increase the consumption of carbomer, its acidity strengthens gradually, and an eye table tear is difficult to be buffered to and changes mutually pH value, and to the corresponding increase of zest of eye, should not be for dosing eyes.

1.3 changing, the ion-sensitive gel phase measures:

Mainly, by relatively adding gel viscosity before and after tear to change, concrete grammar be under 34 ℃ of conditions, repeatedly measures 3 brinzolamide ion-sensitive gels and the mixed rheology variation of different proportion artificial tears.Result shows, can obviously improve the adding of artificial tears viscosity (η) and the elastic modelling quantity (G') of preparation after gelling.Normal eyes table tear amount is about 7 μ L, when the brinzolamide gel mixes with the ratio of 40: 7 with the artificial tears, η and G' than gelling before preparation improve respectively about 2Pas and 5Pa; When mixed proportion reaches 40: 14 and more at high proportion when (40: 32), above-mentioned rheological parameters continues to improve, visible brinzolamide ion-sensitive type gel has good ion-sensitive type gelling ability.

Experimental example 2 galenic pharmacy character are estimated

2.1 the stability of solution of clathrate

The inclusion complex in solution prepared is placed in to room temperature and observes its crystallize time, in order to conclude its stability, prepare for preparing follow-up lyophilized formulations.

The table 2-1 brinzolamide clathrate crystallize time

By table, 2-1 can find out, the stability of brinzolamide inclusion complex in solution is not good, the direct crystallization after preparation completes that the enclose material concentration is less, can't lyophilizing make stable lyophilized formulations, and the enclose material concentration excessive after, can cause very large stimulation to eye, therefore its material concentration is screened, the concentration ratio that the enclose material obtained is 10% to 50% is more suitable.

2.2 the sign of brinzolamide liposome

The laser light scattering particle size analyzer is measured particle diameter and PDI, the preparation made is crossed the filter membrane of 0.22 μ m, on filter membrane for not wrapping into the undissolved medicine of liposome, filtrate is liposome solutions, destroys standardize solution with methanol and chloroform, measure drug level, then be calculated as follows envelop rate E%=C1/C * 100%, C is total concentration, and C1 is the drug level wrapped in liposome.

Table 2-2 brinzolamide liposome

2.3 the sign of brinzolamide nanoparticle

Use laser particle analyzer to measure particle diameter and the PDI of liposome, take a certain amount of nanoparticle lyophilized formulations, add certain DMSO that it is dissolved, high-efficient liquid phase chromatogram determining content, calculate its envelop rate.

Table 2-3 brinzolamide nanoparticle

2.4 the stability of nano suspension

The particle diameter of the nano suspension that mensuration prepares, then add freeze drying protectant, and after lyophilizing, water for injection redissolves, and measures its particle diameter situation.

Table 2-4 brinzolamide nano suspension stability

From table 2-2, brinzolamide nanometer suspension particle diameter is less, and character is more stable, and the preparation after lyophilizing is also more stable, after 45 days, can redissolve easily in aqueous solution, and the preparation change of size is little.

2.5 the stability of pluralgel agent

The preparation for preparing is observed and measure particle diameter after be divided into two parts, portion adds gel-type vehicle to stir, portion does not add gel-type vehicle and stirs, and is placed in room temperature and observes and measure particle diameter after 45 days.

Table 2-5 brinzolamide nanometer suspension gel stores the change of size after 45 days

Annotate: the "/" in table means not need to measure, and embodiment 22 and Comparative Examples 3 are clathrate, do not need to measure particle diameter; In table, "-" means to measure, because it has formed flocculent deposit.

In table 2-5, Comparative Examples is not for adding crystallize time and the change of size of gel, in table, after preparation becomes solution, clathrate and nano suspension are all unstable, crystallize or produce precipitation after 45 days, and jog can't be scattered in solution again, and after adding gel-type vehicle, the stability of preparation increases, produce precipitation or jog after Eddy diffusion in solution, and change of size is little.

Experimental example 3 brinzolamide ophthalmic preparations are exempted from the eye irritation evaluation to family

The research of Ocular irritation research adopts animal consubstantiality left and right sides Self-control method, adopt heavy 2 to 3kg female New Zealand white rabbits to carry out, left eye is administered twice every day, successive administration two weeks, right eye splashes into normal saline and makes blank, after administration, rabbit eyelid passive closed 10 seconds, regularly (before administration every day, observe once, get high score in twice, and after last administration 12 hours, 24 hours, 48 hours, 72 hours) rubescent, swelling and the water-filling situation of observing lagophthalmos section, estimate and undertaken by per Draize technique.Experimental result is in Table 3-1.

Table 3-1 Ocular irritation experimental result

Annotate: 1% brinzolamide eye drop is the aqueous solution that brinzolamide adds 1% citric acid to make, and uses osmotic pressure regulator to adjust osmotic pressure and oozes to waiting.

These standards of grading are to be greater than the preparation of 3 minutes to think larger to Ocular irritation, be less than or equal to the preparation of 3 minutes non-stimulated or zest is very light to eye, table 3-1 shows, brinzolamide citrate zest is very large, and preparation prepared by this patent does not all have zest, demonstrate good eye toleration, also have no the damage of corneal, iris and conjunctiva and abnormal clinical symptoms.

Experimental example 4 brinzolamide gel for eye use tablets in vitro experiments

Adopt bag filter to carry out the extracorporeal releasing test of brinzolamide, by the accurate 1ml formulation samples of drawing of pipette, join in the bag filter of anticipating, after sealing, bag filter is immersed in the 40ml release medium, temperature is controlled at 37 ℃.Release medium for the simulation tear (STF, composed as follows: NaCl0.67g, NaHCO ₃, 0.20g, CaCl ₂.2H ₂o0.008g. distilled water adds to 100ml), with 100rpm rotating speed magnetic agitation.Get 1ml at every turn and discharge sample liquid, and supplementary equal volume simulation tear, discharge sample liquid after 0.45 μ m syringe filter filters, carry out the HPLC analysis, to measure wherein brinzolamide concentration.Result of the test is shown in the comparison of tri-kinds of gel tablets in vitros of Fig. 3 and solution, suspensoid.

Fig. 3 result shows, solution discharges very fast, and suspensoid has certain slow releasing function, and three kinds of gels all can make the steady release of drug slow.The release in vitro of medicine in these polymer supports, mainly by DIFFUSION CONTROLLED, belongs to non-fick diffusion.

Experimental example 5 brinzolamide eyes are exempted from the external permeability of cornea to family with sustained-release gel

Normal rabbits is put to death, separate complete cornea.Cornea is fixed on diffusion cell, cornea inboard (aqueous humor side) is towards acceptance pool, in acceptance pool, acceptable solution is GBR test solution (glutathion sodium bicarbonate green reagent: A liquid: sodium chloride 12.4g/L, potassium chloride 0.716g/L, sodium dihydrogen phosphate 0.206g/L, sodium bicarbonate 4.908g/L, B liquid: calcium chloride 0.230g/L, magnesium chloride 0.318g/L, glucose 1.8g/L, glutathion (oxidized form) 0.184g/L, low tempertaure storage, use front mixed in equal amounts) get embodiment 1, embodiment 4, the brinzolamide eye of embodiment 7 is placed in supply pool with sustained-release gel and suspensoid 0.5g, see through experiment under 37 ℃.With the brinzolamide normal saline solution (TMS) of same concentrations, compare.Get acceptable solution 0.2ml respectively at different time from acceptance pool, supplement the GBR test solution of equal volume.The HPLC method is measured the concentration of brinzolamide in acceptable solution, and calculates the accumulative total transit dose, and experimental result is shown in that tri-kinds of gels of Fig. 4 and solution isolated cornea see through accumulation medication amount result.

As seen from Figure 4, cornea sees through in test in vitro, in same time, the amount that medicine in gel sees through cornea is few than suspensoid, this is mainly because the diffusion velocity of medicine in gel is slow than aqueous solution, and there is stronger electrostatic interaction between medicine and macromolecular material, thereby further delayed the diffusion of medicine and seen through.

Brinzolamide clathrate preparation and the suspensoid of getting embodiment 10, embodiment 11, embodiment 12 with method are placed in supply pool, see through experiment under 37 ℃.With the brinzolamide normal saline solution (TMS) of same concentrations, compare.Get acceptable solution 0.2ml respectively at different time from acceptance pool, supplement the GBR test solution of equal volume.The HPLC method is measured the concentration of brinzolamide in acceptable solution, and calculates the accumulative total transit dose, and experimental result is shown in Fig. 5.Conclusion is: within the specific limits, the amount of enclose material increases, and contributes to medicine to see through cornea, and work as material, surpasses certain amount, saturating cornea is played to retardation.

6 of experimental examples are exempted from eye and are given after the brinzolamide sustained-release gel brinzolamide concentration in tear

Get 6 of normal rabbits, be divided at random two groups, 3 every group.The eye of getting embodiment 1 is administered in the experimental group rabbit eyes with sustained-release gel 50 μ l, and matched group gives the brinzolamide normal saline solution of same concentrations and dosage.Different time after administration, be placed in house with appropriate absorbent cotton and exempt from 30s in a conjunctival sac and dip tear, take tear weight, and, with after the mobile phase eluting, in HPLC method mensuration tear, the concentration of brinzolamide, the results are shown in Table 4.

Table 4 is exempted from eye and is given the brinzolamide concentration in the different time tear after TMS and TMG

From table 4, family exempt from eye give the brinzolamide sustained-release gel after different time after 1 hour, in tear, brinzolamide concentration is significantly higher than the suspensoid of same time, shows that solution can be removed rapidly, and the retention time significant prolongation of gel in eye.

Therefore, ocular in-situ gel preparation of the present invention can improve the bioavailability of brinzolamide at eye, has feasibility.

Claims

1. a brinzolamide ophthalmic preparation, is characterized in that described preparation is selected from the agent of brinzolamide responsive to temperature ocular in-situ gel, the responsive ocular in-situ gel agent of brinzolamide pH, the agent of brinzolamide ion-sensitive ocular in-situ gel, the multiple in-situ gel of brinzolamide, brinzolamide clathrate, the agent of brinzolamide clathrate pluralgel, brinzolamide liposome, the agent of brinzolamide liposome pluralgel, brinzolamide nanoparticle, the agent of brinzolamide nanoparticle pluralgel, brinzolamide nano suspension, the agent of brinzolamide nano suspension pluralgel.

2. brinzolamide ophthalmic preparation according to claim 1, is characterized in that the agent of described brinzolamide responsive to temperature ocular in-situ gel, in the 100ml gel, and the component that contains following weight:

And water is supplied 100ml.

3. brinzolamide ophthalmic preparation according to claim 1, is characterized in that described brinzolamide nanoparticle, in the 100ml nano particle preparations, and the component that contains following weight:

And water is supplied 100ml.

4. brinzolamide ophthalmic preparation according to claim 1, is characterized in that described brinzolamide nano suspension, in the 100ml nano suspension, and the component that contains following weight:

Other pharmaceutically acceptable eye compositions, as:

And water is supplied 100ml.

5. the multiple ocular in-situ gel agent of brinzolamide is characterized in that in the 100ml gel component that contains following weight:

Gel-type vehicle:

And water is supplied 100ml.

6. brinzolamide ophthalmic preparation according to claim 1 is characterized in that described brinzolamide nanoparticle plural gel preparation is in the 100ml nano particle preparations, the component that contains following weight:

Water is supplied 100ml.

7. brinzolamide ophthalmic preparation according to claim 1 is characterized in that described brinzolamide nano suspension plural gel preparation is in the 100ml nano particle preparations, the component that contains following weight:

Other pharmaceutically acceptable eye compositions, as:

Water is supplied 100ml.

8. the application of the brinzolamide ophthalmic preparation of claim 1 in the glaucomatous medicine of preparation treatment.