CN104721145A

CN104721145A - Brinzolamide nanoparticle preparation used for eyes and preparation method thereof

Info

Publication number: CN104721145A
Application number: CN201510113961.8A
Authority: CN
Inventors: 宋相容; 魏于全
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2013-02-21
Filing date: 2013-02-21
Publication date: 2015-06-24

Abstract

The invention relates to the field of medicines and in particular relates to a brinzolamide nanoparticle preparation used for eyes as well as a preparation method and application thereof, aiming to lengthen the residence time of the preparation on the eyes or improve the bioavailability of the preparation. The preparation can be used for treating the diseases, such as intraocular hypertension and glaucoma. Gels can conduce to lengthening the retention time of the medicine on the eyes and reducing the number of times of administration and the dosage of administration, so that not only is the patient compliance improved but also the side effects of the medicine can be reduced. Meanwhile, no preservative is added and single-dosage package is adopted, thus reducing the irritation of eye drops used for a long term to the eyes and greatly improving the medication safety.

Description

A kind of Bu Zuolin amine nanoparticle ophthalmic preparation and preparation method thereof

Technical field

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

Background technology

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

EP2348026 and EP1985618 discloses the synthesis of brinzolamide, US5461081, US5378703, US5240923 and US005948801A disclose the synthesis of brinzolamide and the use as eye drop, wherein there is the prescription of suspensoid, 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 can only suspensoid be made with during other drug conjunctive use, there is very large limitation to its application; JP2010037327 discloses the patent of brinzolamide water formulation, but after medicine salify, its eye holdup time is short, and Corneal trauma is not good, and slow releasing function is poor; WO2009073216A1 proposes the prescription of brinzolamide nano suspension, but needs to make lyophilized formulations, and concerning the glaucoma patient needing medication every day, it is very inconvenient to use; CN2005100434124, its gel-type vehicle is selected from the multivalence hydrochlorate of methylcellulose, is 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 does not propose the concrete prescription of brinzolamide.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 eye retention time in antiseptic life-time service simultaneously, and this area is starved of makes safety, efficient, stable, the controlled liquid preparation facilitating eye to drip by brinzolamide.

Gel for eye use is often carrier with hydrophilic high molecular material, having good biocompatibility, is semi-solid preparation, can action time of prolong drug, reduce administration number of times, can overcome that eye drop bioavailability is low, the shortcoming such as the greasy not easy to apply and suspensoid ophthalmic uncomfortable of Eye ointments.And under the specific physico chemical factor of situ-gel system in conjunctival sac (such as: pH, temperature, ion etc.), self will produce the phase transformation of sol-gel, and become thicker gel by rarer liquid rotating during instillation, because causing the difference of the thixotropic factor of phase in version, be divided into again pH responsive type, responsive to temperature type and ion-sensitive type three kinds.Inventor for developing the in-situ gel of brinzolamide to reach the long-acting control intraocular pressure of slow releasing function, and for providing probability with the use in conjunction of other intraocular pressure lowering medicines, improves the compliance of patient.

Inventor intends using the nano material such as PLGA to prepare brinzolamide nanoparticle, and medicine parcel is become nanometer particle, existence that in aqueous can be stable, and adds its dissolubility in water, makes it have good drug effect.

As mentioned before, inventor has respective feature for the various brinzolamide eye drop formulation implemented, ophthalmic preparation can be made by some specific methods, if the brinzolamide preparation of preparation and gel rubber material can be combined, can solve its stability problem, namely become stable solution and do not need to make lyophilized formulations, or the consumption of stabilizing agent in each preparation can be reduced, or there is the phase in version characteristic of situ-gel.

Summary of the invention

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

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

Ocular in-situ gel agent comprises responsive to temperature ocular in-situ gel agent (being called for short 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)

Disclosed in temperature-sensitive situ-gel, prescription is more, mostly comparatively similar, directly loads brinzolamide be difficult to molding or reach prescription with disclosed prescription, is also difficult to the intraocular pressure lowering effect reaching 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 adopts is certain density methylcellulose and multivalence hydrochlorate composition.One or more combination in any of poloxamer, chitosan mainly selected by thermosensitive hydrogel material of the present invention, and material has had commercial product to obtain, and is convenient to industrialization; Use the brinzolamide thermosensitive hydrogel of the formula preparation of CN2005100434124, it is highly sensitive that its gel phase transition does not have the present invention to announce simultaneously.

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

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 uses glucose phosphate ester.With the brinzolamide thermosensitive hydrogel of this formula preparation, it is highly sensitive that its gel phase transition does not have the present invention to announce.

The embodiment 1 in Penciclovir ophthalmic temperature sensitivity in situ gel preparation and preparation method thereof disclosed in CN200710000552 is adopted to prepare brinzolamide temperature sensing in situ gel rubber, phase transition temperature is 25 DEG C, just gel is formed in storage process, cannot use as eye drop, situ-gel truly can not be formed.Prepare the temperature sensing in situ gel rubber of brinzolamide by eye methazolamide temperature-sensitive situ-gel preparation disclosed in CN200910029314 and preparation method thereof embodiment 3, be also gel in room temperature with regard to phase in version, cannot situ-gel be formed.

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

In 100ml gel, the component containing, for example lower weight:

Brinzolamide or brinzolamide salt are generally 0.1 ~ 10.0g; Preferably 0.1 ~ 5.0g; Most preferably 0.5 ~ 2.0g;

Gel-type vehicle is generally 0.1 ~ 40g; Preferably 5.0 ~ 30g; Most preferably 10 ~ 30g;

Stabilizing agent is generally 0.1 ~ 20g; Preferably 0.1 ~ 15g; Most preferably 0.1 ~ 12g;

Osmotic pressure regulator is generally 0.01 ~ 8.0g; Preferably 0.1 ~ 6.0g; Most preferably 0.1 ~ 4.0g;

PH adjusting agent is generally 0.01 ~ 2.0g; Preferably 0.1 ~ 2.0g; Most preferably 0.1 ~ 1.0g;

And, supply 100ml with water.

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 " usual consumption ", " preferable amount ", " most preferred quantities ".

Thermosensitive hydrogel substrate described in 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 medicine preparation standard, the water in prescription of the present invention all adopts distilled water or water for injection.

In order to prolong drug is in the holdup time of anterior corneal surface, the rate of release of regulating drug, improve the rheological property of gel rubber system, the thickening agent polymer matrix being no more than 5% is added in gel, within the scope of this, osmotic pressure and the viscosity of preparation meet the requirements, the adjuvant concentration increased not only can not improve the character of preparation further, cause the discomfort of patient on the contrary, thickening agent described in 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 thickening agent polymer matrix, be dissolved in water;

B, take thermosensitive hydrogel substrate in proportion again, be dissolved in water, cold preservation is placed to whole dissolving; (illustrate: step B cold preservation used chemistry owned by France, formulation art dissolve the common method of 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 (the degerming mode that available 0.22 μm of membrane filtration or radiation sterilization etc. are conventional, this step is used for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention again to stimulate eye), single dose subpackage and get final product.

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

As one of preferred embodiment of the invention, responsive to temperature ocular in-situ gel of the present invention agent is, in 100ml gel, and the component containing, for example lower 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.Preferred poloxamer molecules weight range is 5000 ~ 20000, and its oxygen ethyl and oxygen propyl group proportion are 4:1 ~ 1:4, most preferably adopts poloxamer188 (F127) and PLURONICS F87 (F68).Preferred chitosan, molecular weight is between 4 ten thousand to 25 ten thousand, 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 the sensitive brinzolamide ocular in-situ gel preparation of phase in version., said preparation in liquid condition, facilitates dropleting medicine-feeding as a solution, is easy to accurate control dose at ambient temperature; Temperature is elevated to and changes gel close to during body temperature in anterior corneal surface, thus increases medicine in the holdup time of eye, delays to eliminate, improves eye bioavailability; Reduce administration number of times, facilitate patient medication.Meanwhile, 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)

Disclosed in pH sensitive in-situ gel, prescription is more, mostly comparatively similar, directly loads brinzolamide be difficult to molding or reach prescription with disclosed prescription.As CN200810116084 discloses Aciclovir eye pH-sensitive in-situ gel and preparation method, what in 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 then carries out a large amount of optimal screening to carbopol and cellulosic consumption, and when using carbopol, its consumption is strict controlled in less than 0.3%, just can guarantee that product final pH can control 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 good pH, and moderate viscosity, be convenient to accurate control dose, facilitate patient to use.Adopt embodiment 1 and embodiment 5 are announced in CN200810116084 prescription to prepare the pH sensitive in-situ gel of brinzolamide respectively, obtained situ-gel viscosity is all too large, is difficult to drip during drop eye drip, cannot accurate control dose.

In 100ml gel, the component containing, for example lower weight:

Gel-type vehicle is generally 0.1 ~ 10.0g; Preferably 0.1 ~ 8.0g; Most preferably 0.1 ~ 6.0g;

And, supply 100ml with water.

The responsive gel-type vehicle of pH described in 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.

Thickening agent described in 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;

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

Cellulose derivative comprises one in methylcellulose, hydroxy methocel, Carboxymethyl cellulose sodium, hydroxypropyl methylcellulose, carboxymethyl cellulose or its combination in any, preferred hypromellose (is called for short HPMC, lower same), preferred employing viscosity is the hypromellose of 1000 ~ 30000cP, if Ka Lekang company standard is the product such as HPMC E4M or HPMC EIOM.Carbopol comprises the one in polyacrylic acid, polyacrylate or CP, preferred CP, commodity are called carbopol, more preferably viscosity is adopted to be the carbopol of 2000-30000cP, if Lubrizol company standard is the products such as carbopol 971, carbopol 980 or carbopol 981.Cellulose derivative and carbopol mixing, as gel-type vehicle, can form polymer interpenetration network gel in aqueous, produce significant stream and become synergism, realize the slow releasing function of brinzolamide.The stream of hypromellose and carbopol becomes synergistic result in table 1.Be used alone hypromellose not there is phase in version characteristic and be used alone the viscosity that carbopol can not reach suitable, therefore the two is mixed 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 the formulation, and be difficult to drip, can not accurately control each dose.

The stream of table 1 hypromellose and carbopol (CP980) becomes synergism

Sample	0.5％HPMC E4M	0.5％HPMC ElOM	1.0％HPMC E4M	0.1％CP 980
					Viscosity (cp, 25 DEG C)	45	102	300	2440
+0.1％CP 980	3975	5355	＞9999

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

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

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 " usual consumption ", " 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.

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

In above-mentioned prescription, in order to improve the stability of preparation, inventive gel agent 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 preferred 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 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, adding excess water mixing, degerming (can adopt the degerming mode that 0.22 μm of filtering with microporous membrane or radiation sterilization etc. are conventional, this step is used for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention again to stimulate eye), obtaining final product.

If interpolation chelating agent, then add in stepb, add in the lump with osmotic pressure regulator and dissolve.

The responsive ocular in-situ gel agent of pH is selected to produce stream and is become synergistic hypromellose and carbopol as mixed gel substrate, two kinds of macromolecular materials are utilized to form polymer interpenetration network gel, prepare 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 makes its pH value increase, and its folding side chain opens, and viscosity increases.Synergism is become because both have significant stream, lower consumption can produce full-bodied gel, thus prolong drug is in the holdup time of eye, on the other hand, positively charged medicine brinzolamide can form complex with electronegative carbopol by electrostatic interaction, thus delay the release of brinzolamide from gel further, produce better slow releasing function, and reduce side effects of pharmaceutical drugs.Meanwhile, gel for eye of the present invention not containing antiseptic, thus to the zest of eye when reducing life-time service, improves drug safety.

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

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

In 100ml gel, the component containing, for example lower weight:

Gel-type vehicle is generally 0.1 ~ 10.0g; Preferably 0.1 ~ 6.0g; Most preferably 0.1 ~ 2.0g;

And, supply 100ml with water.

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

Ion-sensitive gel-type vehicle described in 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 above-mentioned medical hydrophilic high molecular material and thickening agent can be applied with addition of as substrate.

Wherein, gellan gum should be adopted in gel-type vehicle.The complexing of metal ion such as calcium ion, sodium ion, potassium ion in itself and tear forms stable hydrogen bond, and in gel state.Also Yin Wendu and pH value can not change and gelation, facilitate fill.

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

Thickening agent in gel-type vehicle described in 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 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:

By every 100g gel containing 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, pH adjusting agent 0.1 ~ 1.0g, surplus is water preparation.

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

The pH value of final acquisition gel is 4.5 to 9.0.

Use the ion-sensitive type situ-gel prepared by gellan gum isogel substrate, have good ion-sensitive, and supplementary product consumption is few, non-physiological condition current downflow is good, is easy to instill eye, in vitro can sustained release; Alginate is also usually used in preparing 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 ion-sensitive gel, and the phase in version of preparation is responsive, and adjuvant toxic and side effects is little.

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

Multiple gel containing temperature sensitive properties of the present invention, comprises temperature-pH and mixes the responsive multiple in-situ gel of eye, the multiple in-situ gel of eye of temperature-Ar ion mixing sensitivity and the multiple in-situ gel three types of eye of temperature-pH-Ar ion mixing sensitivity.Feature is: this situ-gel morphologically can instill eyes as eye drop, two or three factor be subject in conjunctival sac in temperature, pH of tears or tear intermediate ion is worked in coordination with and is brought out and produce the phase transformation of sol-gel, and when making instillation, rarer liquid rotating becomes thicker gel.Owing to being that multiple factors brings out, phase in version sensitivity can be improved.

It is characterized in that in 100ml gel, the component containing, for example lower weight:

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

Gel-type vehicle:

Responsive to temperature type is generally 0.1 ~ 40g; Preferably 1.0 ~ 30g; Most preferably 1.0 ~ 28g;

Or pH responsive type is generally 0.01 ~ 10.0g; Preferably 0.1 ~ 8.0g; Most preferably 0.1 ~ 6.0g;

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

Or other pharmaceutically acceptable eye compositions are as antiseptic, complexing of metal ion agent etc.;

And, supply 100ml with water.

Present invention also offers one and prepare above-mentioned multiple gel method, combined by two or more phase in version gel formula, be worth the gel preparation with dual or multiple sensitivity, there is better phase in version sensitivity, and increase medicine stability in the formulation, it is characterized in that: A, gel-type vehicle are dissolved in the water of gross weight 25 ~ 35% and obtain 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 nanoparticle and pluralgel agent thereof

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

Although nano particle preparations has lot of documents report, there is no commercialized product as ophthalmic preparation.The present inventor, by adopting specific nano carrier material, while raising brinzolamide dissolubility, substantially increases intraocular pressure lowering therapeutic effect, obtains gratifying technique effect.

CN200610130645 discloses a kind of eye nano eyedrop, use polylactic acid as material, cholesterol modifying chitosan 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, needs to make freeze-dried powder.Release of the present invention is very fast, can enter eye efficiently and reach good antihypertensive effect, and do not need to make lyophilized formulations also can long storage periods.

The object of this invention is to provide a kind of eye nano particle preparations of brinzolamide, this nanoparticle efficient packet can carry brinzolamide, can accumulate and sustained release after eye drip in conjunctival sac; The contact angle of drop and cornea can be reduced, increase spreading coefficient, make preparation have good wettability, thus extend the time of contact of drug-carrying nanometer particle and corneal epithelium; This nanoparticle is also pleasing to the eye by endocytosis, or extends in the holdup time of eye by bio-adhesive effect or electrostatic adsorption, effectively can promote drug absorption, improve drug bioavailability, strengthen drug effect.

The eye nano particle preparations of brinzolamide provided by the invention, stability is better than the nanometer formulation of literature research report, does not therefore need to introduce additional caffolding agent and makes lyophilized formulations.This nanoparticle only pharmaceutical solutions gets final product long storage periods, and eye is with without the need to special device, cost-saving, easy to use.This nanoparticle also can add lyophilizing caffolding agent and make lyophilized formulations.

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

The object of the invention is to provide a kind of eye nano particle preparations, it is characterized in that in 100ml nano particle preparations, the component containing, for example lower weight:

Nanoparticulate materials is generally 0.1 ~ 30.0g; Preferably 0.5 ~ 30.0g; Most preferably 0.5 ~ 20.0g;

Stabilizing agent is generally 0.1 ~ 30.0g; Preferably 0.1 ~ 20.0g; Most preferably 0.1 ~ 15g;

Other pharmaceutically acceptable eye compositions can also be added as antiseptic, complexing of metal ion agent etc.;

And, supply 100ml with water.

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

Nanoparticulate materials described in preparation 4 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, PLGA and derivant thereof, PLA and derivant, PCL and derivant thereof.

Stabilizing agent described in preparation 4 is selected from one or more following combination in any: 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 its derivates, polyoxyethylene castor oil, tyloxapol, sodium alginate, sodium tripolyphosphate, etc..Be preferably polyvinyl alcohol, TPGS, poloxamer and Tween 80; Reason is: these stabilizing agent dosage are relatively little, and obtained nanoparticle particle diameter evenly, stability is better.

Organic solvent used in preparation 4 is selected from following conventional organic reagent, 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 nanoparticle, comprise mon-galacta method, nanoprecipitation method or other can obtain any method of brinzolamide nanoparticle.

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

(1) nanoparticulate materials of recipe quantity and brinzolamide or brinzolamide salt are dissolved in obtained organic facies A in organic solvent;

(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, become uniform Emulsion, or directly form nanoprecipitation granule;

(4) adopt decompression to rotate steaming method or additive method and eliminate organic solvent in Emulsion (if not with an organic solvent, then can save this step), degermingly (the degerming mode that 0.22 μm of filtering with microporous membrane or radiation sterilization etc. are conventional can be adopted, this step is used for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention again to stimulate eye), obtain brinzolamide nanoparticle solution;

(5) or centrifugal or collecting by filtration nanoparticle, carry out lyophilization after again disperseing with the solution containing freeze drying protectant, obtain brinzolamide nanoparticle lyophilized formulations.

The preparation method of preparation method also optional conventional nanoparticle, 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 100ml gel, and the component containing, for example lower weight:

Brinzolamide or brinzolamide salt	0.5～2.0g
		Nanoparticulate materials	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 nanoparticulate carriers material is preferably from: PLGA and derivant, PLA and derivant thereof, Chitosan-phospholipid complex 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, brinzolamide parcel is become nanoparticle, make that its dissolubility in water is the highest improves about 140 times, and the eye liposome gone on the market at present at most only can make dissolubility in brinzolamide water improve about 20 times (extremely unstable of liposome now).Brinzolamide nanoparticle envelop rate is up to more than 85%, and particle diameter is at below 400nm, and colloid solution can steady in a long-term be stored, and stability is better than other nano particle preparations of Liposomal formulation and bibliographical information.Brinzolamide nanoparticle also effectively can promote drug absorption, improves drug bioavailability, strengthens drug effect.

The present invention comprises the art-recognized conventional adjuvant adding other after preparation becomes nanoparticle further and makes other preparation further.5, brinzolamide nanoparticle pluralgel agent (being called for short preparation 5)

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, longer (6 ~ 8h) consuming time, and this not easily means that industry is amplified of ultrafiltration need be used to concentrate nanoparticle; And the position gel pH value made is 4.0 ~ 4.5, meta-acid, has certain zest to eye.Nanoparticle preparation technology of the present invention is simple, short (1 hour) consuming time, and technique is controlled also can amplify production, and pH is moderate, is not with antiseptic, does not have zest to eye.

In 100ml nanoparticle plural gel preparation, the component containing, for example lower weight:

Gel-type vehicle is generally 0.1 ~ 40g; Preferably 0.1 ~ 30g; Most preferably 0.1 ~ 28g;

And, supply 100ml with water.

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

Nanoparticulate materials described in preparation 5 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, PLGA and derivant thereof, PLA and derivant, PCL and derivant thereof.

Gel-type vehicle described in preparation 5 comprises gel rubber material described in preparation 1 to 3 and thickening agent, and the phase in version characteristic of situ-gel can be realized by the prescription proportioning of preparation 1 to 3, 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 gel-type vehicle described in 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.

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

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

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

(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, become uniform Emulsion, or directly form nanoprecipitation granule 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) decompression rotary evaporation or additive method is adopted to eliminate organic solvent in C (if not with an organic solvent, then can save this step), again by C and D mix homogeneously, after adjust ph and osmotic pressure, degermingly (the degerming mode that 0.22 μm of filtering with microporous membrane or radiation sterilization etc. are conventional can be adopted, this step is used for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention again to stimulate eye), obtain brinzolamide nanoparticle plural gel preparation.

Prepare the preparation method of nanoparticle method also optional conventional nanoparticle, as natural polymer polymerization, intra-liquid desiccation method, automatic emulsified method, polymeric bladder Shu Fa etc., after said method obtains brinzolamide nanoparticle solution, the water for injection of surplus is used to dissolve gel-type vehicle, and by after the two mix homogeneously adjust ph and osmotic pressure, degermingly (the degerming mode that 0.22 μm of filtering with microporous membrane or radiation sterilization etc. are conventional can be adopted, this step is used for except degerming, in order to avoid add antiseptic in ophthalmic preparation of the present invention again to stimulate eye), obtain brinzolamide nanoparticle plural gel preparation.

Beneficial effect of the present invention

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

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

Further illustrate:

Brinzolamide salt described in preparation 1 to preparation 5 comprises the salt of the feasible one-tenth of following brinzolamide salt or the arbitrary proportion combination of any kind salt: citrate, maleate, malate, hydrochlorate, acetate, carbonate, bicarbonate, hydrophosphate, phosphoric acid hydrogen disalt, borate etc.

Stabilizing agent described in preparation 1 to preparation 5 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 its derivates, polyoxyethylene castor oil, tyloxapol.

Osmotic pressure regulator described in preparation 1 to preparation 5 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.

The freeze drying protectant of preparation 1 to preparation 5 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 5 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.

PH adjusting agent described in preparation 1 to preparation 5 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.

In preparation 1 to preparation 5, organic solvent used is selected from following conventional organic reagent, comprises one or more combination in any wherein: 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, may be used for disease such as treatment Bulbi hypertonia and glaucoma etc., dosage is generally 1 times/day, and each giving suffers from eye 1, concrete, according to the state of an illness of patient, age etc., can be determined by doctor.Because antiseptic is irritant to eyes, especially serious zest may be caused 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 Bulbi hypertonia patient, and said preparation has long-time slowly release and the feature of foreign sense, can significantly improve the compliance of patient; And comparatively commercial preparation intraocular pressure lowering better effects if, action time more lasting; Meanwhile, preparation used does not add any antiseptic, adopts unit dose package, reduces the zest of eye drop life-time service to eye, substantially increase the safety of medication.This long-acting slow-release preparation technology also can be used for the medicine of other local, realizes slow release long-acting.

Accompanying drawing explanation

Fig. 1 brinzolamide situ-gel 24% poloxamer188 and the viscosity change of 0.5% PLURONICS F87 compatibility tear dilution front and back.

Fig. 2 brinzolamide pH sensitive type gel A, B viscosity B coefficent when different pH value

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

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

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

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

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

Intraocular pressure decreasing value and the commercial preparation of Fig. 6-1 temperature-sensitive situ-gel (embodiment A 6) contrast (" * " represents p<0.05).

Fig. 6-2 temperature-sensitive situ-gel (embodiment A 6) contrasts (" * " represents p<0.05) relative to the intraocular pressure decreasing value of the responsive ocular in-situ gel agent (embodiment A 16) of pH and ion-sensitive ocular in-situ gel agent (embodiment A 25) and commercial preparation.

Intraocular pressure decreasing value and the commercial preparation of the multiple in-situ gel of eye (embodiment A 38) of eye multiple in-situ gel (embodiment A 35), the multiple in-situ gel of eye (embodiment A 36) of temperature-Ar ion mixing sensitivity, the multiple in-situ gel of eye (embodiment A 37) of pH-Ar ion mixing sensitivity and temperature-pH-Ar ion mixing sensitivity that Fig. 6-3 temperature-pH mixing is responsive contrast (" * " represents p<0.05).

Intraocular pressure decreasing value percentage ratio and the commercial preparation of Fig. 7 brinzolamide nanoparticle (Embodiment B 12) contrast (" * " represents p<0.05).

Intraocular pressure decreasing value percentage ratio and the commercial preparation of Fig. 8 nano suspension (Embodiment C 24) contrast (" * " represents p<0.05).

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

Represent p<0.05).

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

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

Detailed description of the invention

Illustrate below by way of specific description of embodiments of the present invention but do not limit the present invention.

Embodiment 1 ~ 3

Note: "/" represents does not add.

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

Embodiment 4 ~ 6

Embodiment 4 to 6 is brinzolamide or brinzolamide salt pH sensitive type gel prescription, hypromellose and carbopol are scattered in soaked overnight in 30ml water for injection by method for making as follows, stir, obtain solution, for subsequent use, by brinzolamide or brinzolamide salt, osmotic pressure regulator, inject and use water 30ml, stir lower dissolving, 0.22 μm of microporous filter membrane filters, in hypromellose filtrate being joined above-mentioned preparation and carbopol solution, benefit injects water to 100ml, adding the pH that qs pH adjuster solution is adjusted to system is 5.5, and stir, obtain brinzolamide gel for eye.

Embodiment 7 ~ 9

Note: "/" represents does not add.

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

Applicant researchs and analyses further above-mentioned three kinds of situ-gels within the priority phase.

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

Note: "/" represents does not add.

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

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

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

Note: "/" represents does not add.

Embodiment A 13 to A20 is brinzolamide or brinzolamide salt pH sensitive type gel prescription, hypromellose and carbopol are scattered in soaked overnight in 30ml water for injection by method for making as follows, stir, obtain solution, for subsequent use, by brinzolamide or brinzolamide salt, osmotic pressure regulator, inject and use water 30ml, stir lower dissolving, 0.22 μm of microporous filter membrane filters, in hypromellose filtrate being joined above-mentioned preparation and carbopol solution, adding water for injection ad pond om is 100ml, adding the pH that qs pH adjuster solution is adjusted to system is 5.5, and stir, obtain brinzolamide gel for eye.

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

Note: "/" represents does not add.

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

The multiple gel preparation of embodiment A 31 ~ A40

Note: "/" represents does not add; "-" expression does 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: added by the macromolecular material of recipe quantity in 20 to 30ml water for injection, magnetic agitation, after being uniformly dispersed, by medicine, osmotic pressure regulator, injects and uses water 30ml, stir lower dissolving, 0.22 μm of microporous filter membrane filters, adjust ph and osmotic pressure, and to add water for injection ad pond om be 100ml, stir, obtain brinzolamide gel for eye.

Experimental example A1 pharmacodynamic evaluation

Get normal rabbits random packet, often organize 6, male and female half and half.Bilateral eye gives 50 μ l Oxybuprocaines (times promise happiness, Japan) eye anesthesia, uses Tono-Pen tonometer measures 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), responsive ocular in-situ gel agent (embodiment A 16) of pH, ion-sensitive ocular in-situ gel agent (embodiment A 25), temperature-pH mixes 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 Pai Liming 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. ) in experimental group rabbit eyes conjunctival sac.After administration 30,60,120,180,240 and measure bilateral varieties of intraocular pressure in 300 minutes minutes, and calculate intraocular pressure decreasing value (mmHg): intraocular pressure decreasing value=certain moment intraocular pressure-Baseline IOP.The inspection of the t in SPSS13.0 software is adopted to carry out statistical procedures, p<0.05 " * " mark.

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

Embodiment 10 ~ 12

Embodiment 10 to 12 is 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 Rotary Evaporators 25 DEG C and boil off organic solvent formation thin film, stabilizing agent is dissolved, add dissolving films, and to add water for injection ad pond om be 100 grams, stir, use pH value regulator and osmotic pressure regulator, adjust ph and osmotic pressure, 0.22 μm of microporous filter membrane filters, filtrate is added freeze drying protectant, stir, with-80 DEG C of pre-freezes 24 hours, lyophilization, obtain brinzolamide clathrate preparation.

Embodiment 13 ~ 15

Note: "/" represents does not add.

Embodiment 13 to 15 is prescriptions of brinzolamide liposome, and method for making is as follows: take the phospholipid of recipe quantity, cholesterol and medicine, dissolve with ethanol, and mix homogeneously mixed liquor is on Rotary Evaporators, and pressure reducing and steaming organic solvent, prepares immobilized artificial membrane; In immobilized artificial membrane, add the water for injection being dissolved with stabilizing agent of 50%, aquation is complete, high pressure homogenize or ultrasonic Treatment adjustment liposomal particle size; Benefit adds to the full amount of water for injection, adjust ph and osmotic pressure, and filtering with microporous membrane, to obtain final product, aseptic subpackaged or add freeze drying protectant and prepare brinzolamide lipidosome freeze-dried preparation.

Embodiment 16 ~ 18

Note: "/" represents does not add.

Embodiment 16 to 18 is prescriptions of preparation 4 brinzolamide nanoparticle, preparation method is as follows: the material of recipe quantity and medicine are dissolved in the organic facies made in dichloromethane, the solution containing stabilizing agent of preparation is aqueous phase, organic facies is poured in aqueous phase, 10000rpm stirs after 3 ~ 5 minutes, in Probe Ultrasonic Searching cell disruptor, ultrasonic 5 minutes of 100W, obtain Emulsion, decompression rotates steams removing organic solvent, 0.22 μm of filtering with microporous membrane is degerming, carries out lyophilization after the sucrose solution with 2% ~ 10% disperses again, obtains brinzolamide nanometer formulation.Applicant studies as follows further to nano particle preparations within the priority phase:

Embodiment B 1 ~ B6 brinzolamide consumption is preferred

Note :=represent suitable with commercially available suspensoid drug effect ,+represent 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, 300W Probe Ultrasonic Searching minutes 5 minutes under condition of ice bath, obtain O/W Emulsion, 37 DEG C of water-bath distilling under reduced pressure eliminate organic solution, adjust ph and osmotic pressure, or add antiseptic, add water and be settled to 100ml, to obtain final product.Intraocular pressure lowering effectiveness comparison is carried out with commercially available suspensoid, result shows, when brinzolamide is 0.05mg/100ml, and drug effect and quite commercially available, improve consumption, drug effect is significantly better than commercially available, and along with concentration increase, drug effect increases gradually, when concentration reaches 2mg/100ml, drug action reaches peak value, increases drug level further, and drug effect no longer includes significant change.

Embodiment B 7 ~ B16 nano carrier material is preferred

Note: "/" represents does not add.

Brinzolamide and nanoparticulate carriers material are dissolved in organic solvent, add in the aqueous solution containing 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 DEG C of water-bath distilling under reduced pressure eliminate organic solution, adjust ph and osmotic pressure, or add antiseptic, add water and be settled to 100ml, to obtain final product.

Embodiment B 17 ~ B22 nano carrier material is preferred

Note: "/" represents does not add.

Mixed with adjuvant by brinzolamide, add in the aqueous solution containing stabilizing agent, high pressure homogenize or dispersant, directly obtain nanoparticle, adjust ph and osmotic pressure, or add antiseptic, adds water and be settled to 100ml, to obtain final product.

Experimental example B1 pharmacodynamic evaluation

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

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

Embodiment 19 ~ 21

Note: "/" represents does not add.

Embodiment 19 to 21 is prescriptions of brinzolamide nano suspension, the preparation method of embodiment 19 and 20: be dissolved in organic facies by medicine and adjuvant, uses high-shear emulsion machine 10000rpm, emulsifying shears 5 minutes, obtain just suspension, high pressure homogenize 300bar, circulates 6 times, 500bar circulates 6 times, 800bar circulates 6 times, and 1000bar circulates 20 times, adjust ph and osmotic pressure and osmotic pressure, cross 0.22 μm of filter membrane, to obtain final product.

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

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 circulates 5 times, and 1000bar circulates 5 times, regulates pH and osmotic pressure, add complexing of metal ion agent and antiseptic, add water and be settled to 100ml, to obtain final product.Intraocular pressure lowering effectiveness comparison is carried out with commercially available suspensoid, result shows, when brinzolamide is 0.05mg/100ml, and drug effect and quite commercially available, improve consumption, drug effect is significantly better than commercially available, and along with concentration increase, drug effect increases gradually, when concentration reaches 2mg/100ml, drug action reaches peak value, increases drug level further, and drug effect no longer includes significant change.

Embodiment C 7 ~ C15 stabilizing agent is preferred

Note: "/" represents does not add.

Brinzolamide is mixed with adjuvant, and be added in the quartzy cup of ball mill with a certain amount of water, use zirconium oxide is abrasive media, 35Hz grinds 6 hours, takes out preparation, crosses and filters abrasive media, adjust ph and osmotic pressure, add complexing of metal ion agent or antiseptic, add water and be settled to 100ml, to obtain final product.Particle size determination result shows, and when making stabilizing agent with glycerol, particle diameter is too large and distributed pole is uneven, and therefore, glycerol is not selected as stabilizing agent.

Note: "/" represents does not add.

Brinzolamide is mixed with adjuvant, and be added in the quartzy cup of ball mill with a certain amount of water, use zirconium oxide is abrasive media, 35Hz grinds 6 hours, takes out preparation, crosses and filters abrasive media, adjust ph and osmotic pressure, add complexing of metal ion agent or antiseptic, add water and be settled to 100ml, to obtain final product; Or add freeze drying protectant, make lyophilized formulations, to obtain final product.Particle size determination result shows, and during with single stable agent, the nano suspension particle diameter that glycerol obtains is too large and distributed pole is uneven, and only can stablize 1 day, 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≤2 month stabilization time of nano suspension, as preferred stabilizing agent.

Experimental example C1 pharmacodynamic evaluation

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

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

Embodiment 22 ~ 25

Note: "/" represents does not add.

Embodiment 22 to 25 is the agent of brinzolamide pluralgel, embodiment 22 prescription is selected from the agent of brinzolamide clathrate pluralgel, 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, organic facies disperse medium A is dripped slowly in B, stir, mixed liquor removing organic solvent, obtain formulation C, use the water dissolution gel-type vehicle of surplus, dissolve completely, by C and gel-type vehicle aqueous solution even, adjust ph and osmotic pressure, 0.22 μm of filtering with microporous membrane is degerming, obtain.

The preparation method of embodiment 23 and 24: be dissolved in organic solvent by the adjuvant such as medicine and suspending agent, obtains organic facies disperse medium A; Stabilizing agent is scattered in the water for injection of recipe quantity, obtains aqueous phase B; In aqueous phase B, drip organic facies disperse medium A slowly, use the physical methods such as mulser to prepare suspensoid colostrum simultaneously, then use the physical method such as Probe Ultrasonic Searching or high pressure homogenize to increase the stability of suspensoid colostrum, obtain suspension emulsion C; Organic solvent in removing C, obtains whole suspensoid D; Use the water for injection of surplus to dissolve gel-type vehicle, stir, obtain gel-type vehicle aqueous solution E, mixed by D and E, adjust ph and osmotic pressure, 0.22 μm of filtering with microporous membrane is degerming, to obtain final product;

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

Note: "/" represents does not add.

The preparation method of brinzolamide compound formulation, comprise the following steps: use the following legal system of water of recipe quantity 50% ~ 80% for nanoparticle: to be dissolved in organic solvent by brinzolamide and nanoparticulate carriers material, add in the aqueous solution containing 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 DEG C of water-bath distilling under reduced pressure eliminate organic solution, then the water of recipe quantity 20% ~ 50% is used to prepare in-situ gel preparation, nanometer formulation is scattered in situ-gel, adjust ph and osmotic pressure, or add antiseptic, obtain.

Embodiment D11 ~ D20 nano suspension situ-gel

Note: "/" represents does not add.

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

Experimental example D1 pharmacodynamic evaluation

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

Obtain result to contrast (" * " represents p<0.05) see Fig. 9-1 nanoparticle solutions suspensions (Embodiment C 24) and nanoparticle ion suspensoid-thermosensitive in situ gel (embodiment D19) intraocular pressure decreasing value, conclusion is after nano suspension and thermosensitive in situ gel compound, 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) contrast (" * " represents p<0.05) with clathrate-pH responsive gel intraocular pressure decreasing value, conclusion is: 3 kinds of compound formulations are by certain intraocular pressure lowering effect, and continuous action time length order is: the responsive gel of nanoparticle gel compound formulation > nanometer suspension gel compound formulation > clathrate-pH; Fig. 9-3 nanoparticle gel compound formulation (embodiment D6), nanometer suspension gel compound formulation (embodiment D19) and commercial preparation contrast (" * " represents p<0.05), and conclusion is: the intraocular pressure lowering effect of nanoparticle gel compound formulation and nanometer suspension gel compound formulation is all better than commercial preparation.

The phase in version of experimental example 1 brinzolamide temperature, pH and ion-sensitive gel for eye use detects

1.1 temperature sensitive hydrogel phase transition temperatures measure:

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

Brinzolamide gel preparation gel phase transition temperature measurement result (n=3) of the different prescription of table 1-1

Table 1-1 result shows: add the gelation temperature that hypromellose can reduce Poloxamer 127 to a certain extent.Compared with the Poloxamer 127 of alone same concentrations, 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 increased the ratio of Poloxamer 127, gel phase transition temperature reduces, and higher to the conditional request of producing, transporting and storing, cost increases, and use inconvenience, and reducing the ratio of Poloxamer 127, its phase transition temperature raises, and the temperature of eyeball surface is 34 DEG C to 37 DEG C, the requirement of its phase in version can not be met, cannot gel be formed.

The mensuration of 1.2pH sensitive type gel phase in version pH value:

At 34 ± 0.2 DEG C, the viscosity that pH value is 4.0-9.0 gel when measuring this pH value is set respectively, becomes suddenly large point as phase in version pH point using its viscosity, as shown in Figure 2.

Different prescription brinzolamide gel preparation gel phase transition pH measurement result (n=3) of table 1-2

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

1.3 ion-sensitive gel phase transition measure:

Add gel viscosity change before and after tear mainly through comparing, concrete grammar is under 34 DEG C of conditions, and mensuration 3 brinzolamide ion-sensitive gels and the mixed rheology of different proportion artificial tears change repeatedly.Result shows, can significantly 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 brinzolamide gel and artificial tears with 40: 7 ratio mix time, η and G' improves about 2Pas and 5Pa respectively than preparation before gelling; When mixed proportion reach 40: 14 and more at high proportion (40: 32) time, above-mentioned rheological parameters continues to improve, and visible brinzolamide ion-sensitive type gel has good ion-sensitive type gelling ability.

Experimental example 2 galenic pharmacy properties evaluations

The sign of 2.1 brinzolamide nanoparticles

Table 2-1 brinzolamide nanoparticle

The stability of 2.2 pluralgel agent

Observed by the preparation prepared and be divided into two parts after mensuration particle diameter, portion adds gel-type vehicle and stirs, and portion does not add gel-type vehicle and stirs, and is placed in room temperature and observes after 45 days and measure particle diameter.

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

Note: the "/" in table represents not to be needed to measure, and namely embodiment 22 and comparative example 3 are clathrate, does not need to measure particle diameter; In table, "-" expression cannot measure, because it forms flocculent deposit.

In table 2-2, comparative example is the crystallize time and the change of size that do not add gel, from in table, after preparation becomes solution, clathrate and nano suspension are all unstable, and after 45 days, crystallize or generation precipitate, and jog cannot be scattered in solution again, and after adding gel-type vehicle, the stability of preparation increases, and after not producing precipitation or jog, Eddy diffusion is in solution, and change of size is little.

Experimental example 3 brinzolamide ophthalmic preparation exempts from eye irritation evaluation to family

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

Table 3-1 Ocular irritation experimental result

Note: 1% brinzolamide eye drop is the aqueous solution that brinzolamide adds 1% citric acid and makes, uses osmotic pressure regulator to adjust osmotic pressure to isotonic.

These standards of grading are that the preparation being greater than 3 points thinks larger to Ocular irritation, the preparation being less than or equal to 3 points is non-stimulated or zest is very light to eye, table 3-1 display, brinzolamide citrate zest is very large, and preparation prepared by this patent does not all have zest, demonstrate good Ocular Tolerability, also have no the damage of corneal, iris and conjunctiva and the clinical symptoms of exception.

Experimental example 4 brinzolamide gel for eye use tablets in vitro is tested

Adopt bag filter to carry out the extracorporeal releasing test of brinzolamide, draw 1ml formulation samples by pipette precision, join in the bag filter of anticipating, after sealing, immersed by bag filter in 40ml release medium, temperature controls at 37 DEG C.Release medium for simulation tear (STF, composed as follows: NaCl 0.67g, NaHCO ₃, 0.20g, CaCl ₂.2H ₂o 0.008g. distilled water adds to 100ml), with 100rpm rotating speed magnetic agitation.Get 1ml at every turn and discharge sample liquid, and supplement same volume simulation tear, release sample liquid, after 0.45 μm of syringe frit, carries out HPLC analysis, to measure wherein brinzolamide concentration.Result of the test is shown in Fig. 3 tri-kinds of gel tablets in vitros and solution, the comparing of suspensoid.

Fig. 3 result shows, solution release is 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 controls primarily of diffusion, belongs to non-fick and spreads.

Experimental example 5 brinzolamide eye sustained-release gel exempts from the external permeability of cornea to family

Normal rabbits is put to death, is separated complete cornea.Cornea is fixed on diffusion cell, inside cornea, (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) Example 1, embodiment 4, brinzolamide eye sustained-release gel and the suspensoid 0.5g of embodiment 7 are placed in supply pool, carry out through experiment at 37 DEG C.Compare with the brinzolamide normal saline solution (TMS) of same concentrations.From acceptance pool, get acceptable solution 0.2ml respectively at different time, supplement the GBR test solution of same volume.HPLC method measures the concentration of brinzolamide in acceptable solution, and calculates accumulative transit dose, and experimental result is shown in that Fig. 4 tri-kinds of gels and solution isolated cornea are through accumulation medication amount result.

As seen from Figure 4, in vitro in cornea permeability test, in same time, the amount of the agent permeates therethrough cornea in gel is few compared with suspensoid, mainly because of the diffusion velocity of medicine in gel, comparatively aqueous solution is slow for this, and there is between medicine and macromolecular material stronger electrostatic interaction, thus delayed further medicine diffusion and through.

Be placed in supply pool with the brinzolamide clathrate preparation of method Example 10, embodiment 11, embodiment 12 and suspensoid, carry out through experiment at 37 DEG C.Compare with the brinzolamide normal saline solution (TMS) of same concentrations.From acceptance pool, get acceptable solution 0.2ml respectively at different time, supplement the GBR test solution of same volume.HPLC method measures the concentration of brinzolamide in acceptable solution, and calculates accumulative transit dose, and experimental result is shown in Fig. 5.Conclusion is: within the specific limits, and the amount of enclose material increases, and contributes to agent permeates therethrough cornea, and when material exceedes certain amount, then plays retardation to saturating cornea.

Experimental example 6 is exempted from eye and to be given after brinzolamide sustained-release gel brinzolamide concentration in tear

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

The brinzolamide concentration that eye gives after TMS and TMG in different time tear exempted from by table 4

From table 4, family exempt from eye give 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 rapidly cleared in humans, 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 brinzolamide nanoparticle, nanoparticle plural gel preparation.

2. brinzolamide ophthalmic preparation according to claim 1, is characterized in that described brinzolamide nanoparticle, in 100g nano particle preparations, and the component containing, for example lower weight:

Brinzolamide or brinzolamide salt are generally 0.10 ~ 10.0g; Preferably 0.1 ~ 5.0g; Most preferably 0.5 ~ 2.0g;

Stabilizing agent is generally 0.10 ~ 30.0g; Preferably 0.1 ~ 20.0g; Most preferably 0.1 ~ 15g;

And other pharmaceutically acceptable eye compositions, as:

Osmotic pressure regulator is appropriate, makes osmotic pressure be 280 ~ 320 Osmol/kg;

PH adjusting agent is appropriate, makes pH value be 5.0 to 9.0;

Other pharmaceutically acceptable eye compositions can also be added as antiseptic etc.;

Finally, 100g is supplied with water.

3. brinzolamide ophthalmic preparation according to claim 1, is characterized in that described nanoparticle plural gel preparation, in 100ml nanoparticle plural gel preparation, and the component containing, for example lower weight:

And, supply 100ml with water.

4. the brinzolamide ophthalmic preparation according to Claims 2 or 3, it is characterized in that nanoparticulate materials is selected from following material, comprise one or more the combination in any in following material: albumin, gelatin, chitosan, PACA and derivant thereof, PMMA and derivant, PLGA and derivant thereof, PLA and derivant, PCL and derivant thereof.

5. a preparation method for brinzolamide nano particle preparations, is characterized in that comprising the following steps:

6. a preparation method for brinzolamide nanoparticle compound formulation, is characterized in that comprising the following steps:

7. the application of brinzolamide ophthalmic preparation in the glaucomatous medicine of preparation treatment of claim 1-3.