Carrier or auxiliary material of ophthalmic preparation and preparation method and application thereof

文档序号：1968062 发布日期：2021-12-17 浏览：5次中文

阅读说明：本技术 一种眼用制剂的载体或辅料及其制备方法和应用 (Carrier or auxiliary material of ophthalmic preparation and preparation method and application thereof ) 是由董庆张舒成旋薛陆兵唐欣于 2021-06-17 设计创作，主要内容包括：本发明提供了一种眼用制剂的载体或辅料及其制备方法和应用,属于眼用药递送领域。本发明眼用制剂的载体或辅料的含有如下成分：表面活性剂和离子型高分子等,还含有溶剂；本发明眼用制剂的载体或辅料中含有所述成分在溶剂中形成的纳米小体,纳米小体为球形,粒径为1～100nm,可自组装形成粒径为10～2000nm的球形的纳米小球。本发明给药载体能够包裹药物穿过眼前段,将药物高效输送到眼后段发挥治疗作用,实现了通过滴眼给药治疗眼底疾病的目标,解决了眼科药品给药领域一直努力解决但未解决的技术问题,具有极为优良的临床使用前景和非常积极的社会意义。(The invention provides a carrier or auxiliary material of an ophthalmic preparation, a preparation method and application thereof, and belongs to the field of ophthalmic drug delivery. The carrier or the auxiliary material of the ophthalmic preparation contains the following components: surfactant, ionic polymer, etc., and further contains solvent; the carrier or the auxiliary material of the ophthalmic preparation contains a nano corpuscle formed by the components in a solvent, wherein the nano corpuscle is spherical, has the particle size of 1-100 nm, and can be self-assembled to form spherical nano globules with the particle size of 10-2000 nm. The drug delivery carrier can wrap the drug to pass through the anterior segment of the eye, and efficiently deliver the drug to the posterior segment of the eye to play a therapeutic role, thereby realizing the aim of treating fundus diseases by eye drop drug delivery, solving the technical problems which are always solved but not solved in the field of ophthalmic drug delivery, and having extremely good clinical application prospect and very positive social significance.)

1. A carrier or adjuvant for an ophthalmic preparation, characterized in that it comprises the following components: surfactant, ionic polymer and solvent.

2. The carrier or adjuvant for ophthalmic preparations according to claim 1, wherein the surfactant and ionic polymer are in a mass ratio of: (1-100) 0.1-50; the proportion of the surfactant to the solvent is as follows: 5-3000 mg of surfactant is contained in each 100mL of solvent;

preferably, the mass ratio of the surfactant to the ionic polymer is (1-31): 1-7.5); the proportion of the surfactant to the solvent is as follows: each 100mL of the solvent contains 50-3000 mg of the surfactant.

3. A carrier or adjuvant for an ophthalmic formulation according to claim 1 or 2, characterized in that: the surfactant is a nonionic surfactant.

4. A carrier or adjuvant for an ophthalmic formulation according to claim 3, characterized in that: the non-ionic surfactant is span, polysorbate, poloxamer, alkyl glucoside, vitamin E polyethylene glycol succinate, sucrose stearate or azone.

5. A carrier or adjuvant for an ophthalmic formulation according to claim 1 or 2, characterized in that: the ionic polymer is selected from carboxymethyl cellulose and its salt, sodium starch glycolate, hyaluronic acid and its salt, xanthan gum, alginic acid and its salt, and polyethylene glycol diacetate PEG- (COOH)₂At least one of (1).

6. A carrier or adjuvant for an ophthalmic preparation, characterized in that it comprises the following components: low-polymerization degree polyvidone and medium-polymerization degree polyvidone, and also contains solvent.

7. The vehicle or excipient for ophthalmic preparations according to claim 6, wherein the mass ratio of the low-polymerization degree povidone to the medium-polymerization degree povidone is: (0.1-10) 1, wherein the ratio of the low-polymerization-degree povidone to the solvent is as follows: 5-3000 mg of low-polymerization-degree povidone is contained in every 100mL of the solvent;

preferably, the mass ratio of the low-polymerization degree povidone to the medium-polymerization degree povidone is as follows: (0.24-1) 1, wherein the ratio of the low-polymerization-degree povidone to the solvent is as follows: every 100mL of the solvent contains 400-840 mg of low-polymerization-degree povidone.

8. The vehicle or excipient for ophthalmic preparations according to claim 6 or 7, wherein the low-polymerization degree povidone is povidone having a weight average molecular weight of 2000 to 5000Dalton, and the medium-polymerization degree povidone is povidone having a weight average molecular weight of 20000 to 60000 Dalton.

9. The vehicle or excipient for ophthalmic preparations according to claim 6 or 7, wherein said low-polymerization degree povidone is povidone PVP K12 having a weight average molecular weight of 3500Dalton, and said medium-polymerization degree povidone is povidone PVP K30 having a weight average molecular weight of 35000 to 50000 Dalton.

10. A carrier or adjuvant for an ophthalmic formulation according to any one of claims 1 to 9, characterised in that: the solvent is a polar solvent.

11. A carrier or adjuvant for an ophthalmic formulation according to claim 10, characterized in that: the polar solvent is water.

12. A carrier or adjuvant for an ophthalmic formulation according to any one of claims 1 to 11, characterized in that: it also contains the following components: tackifiers and/or cosolvents;

preferably, the tackifier is at least one of polyethylene glycol, carbomer, poloxamer, povidone, hydroxypropyl cellulose, hydroxyethyl cellulose, methyl cellulose, polyvinyl alcohol, xanthan gum, polyoxyethylene fatty alcohol, hyaluronic acid and salt thereof or hydroxypropyl methyl cellulose, and the cosolvent is propylene glycol, glycerol, liquid polyethylene glycol, hydrogenated castor oil or castor oil; the mass ratio of the tackifier to the surfactant is 1 (0.1-100), and the mass ratio of the cosolvent to the surfactant is 1 (1-10) to 1; the mass ratio of the tackifier to the low-polymerization-degree povidone is 1 (0.1-100), and the mass ratio of the cosolvent to the low-polymerization-degree povidone is 1 (1-10) to 1;

more preferably, the mass ratio of the tackifier to the surfactant is 1 (0.1-30); the mass ratio of the tackifier to the low-polymerization-degree povidone is 1 (0.1-30).

13. The vehicle or adjuvant for ophthalmic preparations according to any one of claims 1 to 12, characterized in that it comprises nanocsomes having a spherical shape and a particle size of 1 to 100 nm; the nanometer corpuscle is formed by self-assembling components of a carrier or an auxiliary material of the ophthalmic preparation.

14. A carrier or adjuvant for an ophthalmic formulation according to claim 13, characterized in that: the particle size of the nano particles is 5-30 nm.

15. A carrier or adjuvant for an ophthalmic formulation according to claim 13 or 14, characterized in that: the nano-microsphere is spherical, and the particle size of the nano-microsphere is 10-2000 nm; the nanospheres are formed by self-assembly of nanospheres.

16. A carrier or adjuvant for an ophthalmic formulation according to claim 15, characterized in that: the particle size of the nano-spheres is 100-2000 nm.

17. A method for preparing a carrier or adjuvant for an ophthalmic preparation according to any one of claims 1 to 16, characterized in that: the preparation method comprises the steps of uniformly mixing the components and a solvent into a solution, and then grinding or uniformly dispersing the solution.

18. Use of a carrier or adjuvant for an ophthalmic formulation according to any one of claims 1 to 16 in the manufacture of a vehicle for eye drop administration for delivery of a drug to the posterior segment of the eye.

19. An ophthalmic preparation for eye drop administration, characterized in that: the ophthalmic preparation is a preparation consisting of the carrier or the auxiliary material of the ophthalmic preparation as described in any one of claims 1 to 16 and an active ingredient for treating eye diseases.

20. The ophthalmic preparation as claimed in claim 19, wherein the carrier or adjuvant of the ophthalmic preparation comprises a surfactant and an active ingredient for treating eye diseases in a mass ratio of (1-30) to (1-2);

or the mass ratio of the low-polymerization-degree povidone in the carrier or the auxiliary material of the ophthalmic preparation to the active ingredient for treating the eye diseases is (6-40) to 1.

21. The ophthalmic formulation of claim 19, wherein: the carrier or the auxiliary material of the ophthalmic preparation contains nano-particles, and the active component for treating the eye disease is wrapped in the nano-particles.

22. The ophthalmic formulation of claim 21, wherein: the nano corpuscle is spherical, the particle size of the nano corpuscle is 1-100 nm, and preferably, the particle size of the nano corpuscle is 5-30 nm.

23. An ophthalmic formulation according to claims 19 to 22, characterized in that: the active component for treating the eye diseases comprises small molecule compound medicines, or free acids, or free bases, or pharmaceutically acceptable salts thereof;

preferably, the small molecule compound medicines comprise nucleoside antiviral medicines, ocular hypotensive medicines, antibiotic medicines, antioxidant medicines, anti-inflammatory medicines, muscarinic receptor blocker medicines, immunosuppressant medicines and glucocorticoid medicines;

more preferably, the nucleoside antiviral drugs comprise ganciclovir, acyclovir, penciclovir, cidofovir, fosmivir and lobucavir;

the ocular hypotensive drug comprises carbonic anhydrase inhibitor, more preferably brinzolamide, acetazolamide, methazolamide;

the antibiotic medicines comprise amikacin, ceftriaxone, cefazolin, oxacillin, levofloxacin, ciprofloxacin, moxifloxacin and vancomycin;

the anti-inflammatory drugs include: oxytetracycline;

the antioxidant comprises taurine, anthocyanin and lignin;

the muscarinic receptor blocker drugs comprise atropine, scopolamine and anisodamine;

the immunosuppressant drugs comprise: cyclosporin (cyclosporine), tacrolimus (tacrolimus), sirolimus (sirolimus), everolimus (everolimus), mycophenolate mofetil (mycophenolate mofetil), methotrexate, azathioprine, and cyclophosphamide;

the glucocorticoid medicine comprises cortisone, prednisone, prednisolone, methylprednisolone, triamcinolone acetonide.

24. A method of preparing the ophthalmic preparation of any one of claims 19 to 23, characterized in that: the method comprises the following steps:

(1) adding a surfactant and/or a tackifier into a solvent to prepare a solution;

(2) dispersing active ingredients and/or cosolvent for treating eye diseases in the solution obtained in the step (1), adding ionic polymer or the solution thereof, and dispersing and mixing to obtain primary suspension;

(3) stirring and dispersing or homogenizing and dispersing the primary suspension obtained in the step (2) to obtain the suspension;

or comprises the following steps:

(a) adding low-polymerization-degree povidone and/or tackifier into a solvent to prepare a solution;

(b) dispersing active ingredients and/or cosolvent for treating eye diseases in the solution obtained in the step (a), adding povidone with medium polymerization degree or solution thereof, and dispersing and mixing to obtain primary suspension;

25. The method according to claim 24, wherein the dispersion in step (2) or step (b) is at least one selected from the group consisting of mechanical stirring dispersion, magnetic stirring dispersion, vortex shaking dispersion, shear dispersion, homogeneous dispersion, milling dispersion and ultrasonic dispersion.

26. Use of an ophthalmic formulation according to any one of claims 19 to 23 in the manufacture of a medicament for the treatment of an ocular disease.

27. Use according to claim 26, characterized in that: the ophthalmic preparation is an ophthalmic preparation for treating fundus diseases and/or treating viral infectious diseases of the posterior segment of the eye and/or treating chronic inflammation of the posterior segment of the eye and/or reducing intraocular pressure and/or eye pain and/or resisting bacterial or fungal infection in the eye, and/or an ophthalmic preparation for preventing and treating juvenile myopia and pseudomyopia and/or an ophthalmic preparation for treating autoimmune diseases and/or an ophthalmic preparation for treating diseases of the anterior segment of the eye and/or an ophthalmic preparation for inhibiting tumor growth.

28. Use according to claim 27, characterized in that:

the ophthalmic preparation for treating fundus diseases comprises ophthalmic preparations for treating macular edema, inflammatory edema and inflammatory pain caused by fundus vascular diseases; more preferably, the ophthalmic preparation is used for treating central retinal vein obstructive macular edema, branch retinal vein obstructive macular edema, diabetic retinopathy, diabetic macular edema, pathologic myopia macular edema, macular edema caused by wet age-related macular degeneration, dry macular degeneration, geographic atrophy, endophthalmitis, acute retinal necrosis, postoperative inflammatory pain, uveitis;

the ophthalmic preparation for treating the virus infectious diseases of the posterior segment of the eye comprises ophthalmic preparations for treating cytomegalovirus uveitis, viral optic neuritis and viral acute retinal necrosis;

the ocular preparation for reducing intraocular pressure comprises an ocular preparation for treating acute and chronic glaucoma and complications thereof;

the ophthalmic preparation for treating the autoimmune disease is an ophthalmic preparation for treating an ocular disease caused by an ocular immune disease or a systemic autoimmune disease, and preferably comprises an ophthalmic preparation for treating Graves eye disease, Behoet's syndrome, shotgun projectile retinal choroidopathy, sicca syndrome, sympathetic ophthalmia or granulomatous eye disease;

the ophthalmic preparation for treating the disease of the front of the eye comprises an ophthalmic preparation for treating sequelae or complications after high-risk corneal transplantation, vernal catarrhal keratoconjunctivitis, silkworm-erosive corneal ulcer or refractory corneal ulcer, herpes simplex viral keratitis, corneal neovascularization and corneal pterygium.

29. Use according to claim 27, characterized in that the active ingredient of the ophthalmic preparation against intraocular bacterial or fungal infections is an antibiotic;

the active ingredient of the ophthalmic preparation for preventing and treating juvenile myopia and pseudomyopia is a muscarinic receptor blocker;

the active ingredient of the ophthalmic preparation for treating autoimmune diseases is an immunosuppressant.

Technical Field

The invention belongs to the field of ophthalmic drug delivery, and particularly relates to a carrier or auxiliary material of an ophthalmic preparation, and a preparation method and application thereof.

Background

The number of patients with fundus diseases is large, the number of Chinese patients is more than tens of millions, and with the increasing aging of society and the popularization of electronic products, the incidence of diseases will rise year by year; common fundus diseases comprise diabetic macular edema, diabetic retinopathy, age-related macular degeneration, retinal vein occlusion, pathological myopia, geographic atrophy, eye tumor, endophthalmitis and the like, can cause vision loss and even blindness, and seriously affect the life quality of people. For example, about 6.8% of diabetic patients suffer from Diabetic Macular Edema (DME), which is the leading cause of blindness in diabetic patients (Urias et al, Vision Research, V139:221-227, 2017; Manual et al, Ocular Delivery of proteins and peptides: changes and novel recommendation, Advanced Drug Delivery Reviews,126: 67-95,2018).

Drug therapy is the main treatment method and research trend for fundus diseases. However, due to the complex physiological structure and barriers in the eye, it is difficult to get the drug into the eyeball, especially the posterior segment of the eye, and to achieve an effective dose, and thus to achieve effective treatment. The search for an effective and safe medicine or method for treating fundus diseases is an struggle goal of cumin which researchers in the field are always seeking.

Clinical ophthalmic administration is usually by 3 routes: first conjunctival sac drug delivery (eye drop): the medicine can be diffused and distributed to iris and ciliary body after entering into the aqueous humor of the anterior chamber through the cornea, but the medicine is difficult to enter the lens and the vitreous body due to the barrier effect of the lens and the vitreous body membrane; the injection for the eyes is as follows: including subconjunctival injection, anterior chamber injection, vitreous injection, retrobulbar injection and orbital injection, the injection can make the drug reach the treatment site directly, but the injection is traumatic and has potential risks, such as pain caused by anterior chamber injection, photophobia, lacrimation, anterior chamber turbidity, hemorrhage, corneal endothelial cell damage, traumatic cataract and the like; vitreous injections can cause lenticular opacification, vitreous organisation, retinal/optic nerve damage, etc.; systemic administration includes oral administration, intravenous administration: most of the drugs in vivo generally accumulate in the liver, kidney or lung, and are hindered by the blood-retinal barrier (BRB), so that the concentration of the drugs reaching the eyeball tissue is low, and unnecessary toxic and side effects are borne by the whole body, especially major organs.

Currently, in order to make drugs pass through the ocular barrier, technical means such as vitreous injection or vitreous implantation (intraocular insertion) are generally adopted to deliver drugs to the vitreous of a patient to treat diseases of the posterior segment of the eye (the ministry of pediatrics, ophthalmic drugs and Pharmaceutics, light industry press, 2010, P3; Wang et al, Mediators of Inflammation, vol.2013, Article ID 780634; luces-Rodr i guez et al, Pharmaceutics, 2018,10, 66). The operation of vitreous injection or vitreous implantation of drugs is traumatic administration and requires specially trained ophthalmologists to operate in aseptic environments such as operating rooms; complications such as ocular hypertension, cataract, iatrogenic infectious endophthalmitis, vitreous hemorrhage, retinal damage, etc. may occur due to traumatic manipulation; the operating conditions and the operating environment have high requirements and need to be carried out in a conditional hospital; the production cost and the use cost of the biological medicine ophthalmic injection are high; meanwhile, the treatment is delayed due to the limitation of medical conditions on the treatment occasion, and the flexibility of adjusting the administration scheme is poor (M.HATA et al, RETINA, 37: 1320-.

Conjunctival sac administration is the most convenient and safest way of ocular administration. However, the cornea has a multilayer structure, which is roughly divided into: the eye drops are first contacted with the epiphora aqueous layer after being dropped into the eye, and then need to cross the epithelial layer, the basal layer and the endothelial layer to reach the posterior segment of the eye. In this process, eye drops tend to be concentrated in the tissues of the anterior segment of the eye due to tear dilution, ocular surface barrier of the cornea, conjunctiva and anatomical location of the lens, vitreous body, and it is difficult to reach the posterior segment of the eye and achieve effective therapeutic concentrations. Therefore, the mode of conjunctival sac administration is safe, but the drug delivery is poor, and it is difficult to achieve the purpose of effectively treating the ocular fundus disease.

In conclusion, the existing main drug administration methods for treating fundus diseases are difficult to realize safety and effectiveness.

Compared with intravenous injection and vitreous injection, the eye drop administration mode has the obvious advantages of safety and convenience, and the ophthalmic preparation capable of delivering the medicine to the posterior segment of the eye is a technical problem to be solved urgently in clinical practice and has clinical treatment value and social significance. However, due to the particularity of the eyeball structure, the multiple barriers of the water-soluble layer and the lipid-soluble layer must be passed through for multiple times when entering the posterior segment of the eye, and at present, researchers have made various attempts, but all of them do not achieve the ideal effect.

Nanotechnology allows drugs to be dispersed to the nanometer scale with special physicochemical properties and different Drug distribution absorption characteristics, increasing tissue and cell penetration, potentially leading to new therapeutic effects for drugs (T.L. CHANG et al, Nanocrytal technology for Drug Delivery and Delivery, front.chem.Sci.Eng.2015,9(1): 1-14; S.Jiang et al, Nanocrytal in Delivery, int.J. Ophthalmol., 2018,11(6)1038 + 1044; M.Kabiri et al, Delivery-Delivery, in situ-formation, nanoparticule-layer hydrogel for tissue Delivery, Drug Delivery Research,2018, 20199, et al: new tissue and cell Delivery materials Research, 2019, 99, et al). For example, nanotechnology is used in the preparation of ophthalmic drugs to increase bioavailability and controlled release delivery. The eye drops such as hydrocortisone nanosuspensions (particle size: 0.539-4.87 μm) prepared by Kassem et al can increase the intraocular pressure of rabbits (M.A. Kassem et al, Nanosuspension as an ocular delivery system for cardiac opthalmic delivery drugs, International J.pharmaceuticals, 340(2007): 126-. Studies have reported that nanocapsules of cyclosporin a can enter the surface layer of the cornea of experimental animals, but fail to enter the whole layer of the cornea; for another example, studies report that after PEG polycaprolactone nanoparticles are dropped into rabbit eyes, the nanoparticles can penetrate into corneal epithelium; it has also been found that the controlled and delayed release of the drug can be achieved by subconjunctival and intravitreal injection of the nano-drug (jin Yi Guang eds., "application of nanotechnology in drug delivery", P319, chemical industry Press 2015). The non-ionic surfactant/nonionic amphiphilic compound (such as span, poloxamer, tween and the like) is matched with cholesterol to coat the medicine to form vesicles (NSVs, also called niosomes) with the diameter of mostly submicron, which are used as novel medicine delivery carriers. The biological adhesive material can enhance the retention time of the vesicle in the eye, maintain the slow release of the drug, reduce the clearance of the drug in the eye, and the like, and can enhance the penetration to the cornea. Improvement of Drug bioavailability in animal experiments with Vesicles prepared with cyclopentamates (mydriatic), timolol maleate (glaucoma Drug) and acetazolamide (glaucoma Drug), respectively (dawn zhao et al, progress in the Application study of novel Drug carrier Non-Ionic Surfactant Vesicles, chinese hospital pharmaceutical journal, 2008, 28 (1): 833-5; x.ge et al, Advances of n-Ionic Surfactant Vesicles (Niosomes) and therapeutic Application in Drug Delivery, pharmaceuticals, 2019,11, 55). Puras et al injected prepared cationic vesicles into rat retina and subretina as a means of gene delivery (G.Puras et al, A novel cationic liposome formation for gene delivery to the retina, J.Control.Release, 2014,174, 27-36). Although the preparation of ophthalmic drugs using nanotechnology has certain advantages, the above studies have not yet achieved the goal of delivering drugs to the posterior segment of the eye by non-invasive eye drop delivery and ensuring effective drug concentration.

The Nano-Emulsion (Nano-Emulsion) is also called microemulsion and consists of an oil phase, a water phase, a surfactant and a cosurfactant according to a proper proportion, can have both hydrophilic and lipophilic characteristics, and has the potential possibility of preparing a drug delivery system for delivering drugs to the posterior segment of eyes by eye drop administration. The conventional nanoemulsion is difficult to have good hydrophilicity and lipophilicity simultaneously, so that the nanoemulsion can sequentially pass through an ocular surface lacrimal fluid layer, a corneal epithelial cell layer, a solid layer and an endothelial cell layer and then deliver an effective drug into the posterior segment of an eye. Vegetable oil is often adopted in the oil phase of the nanoemulsion, and the oil ester is decomposed, so that the stability of the preparation is possibly poor; moreover, the used surfactant is large in dosage (25-30%), and toxic and side effects and anaphylactic reactions can be caused; cosurfactants such as ethanol, which increase osmotic pressure, are not suitable for eye drops, and are unstable (ed by jinyi light, application of nanotechnology in drug delivery, P322, chemical industry press, 2015; ed by panacea, pharmaceutics, P404, chemical industry press, 2017).

Therefore, the research on an eye drop administration vehicle for effectively delivering a drug to the posterior segment of the eye remains a technical problem to be solved and not solved in the art.

Disclosure of Invention

In view of the above problems, the present invention provides an eye drop delivery vehicle for efficiently delivering a drug to the posterior segment of the eye and an application thereof, and the eye drop delivery vehicle of the present invention can efficiently deliver the drug to the posterior segment of the eye, thereby solving the technical problem that people in the field of ophthalmic drug delivery have been eagerly solved but have not been successful all the time.

The invention provides a carrier or an auxiliary material of an ophthalmic preparation, which comprises the following components: surfactant, ionic polymer and solvent.

Further, the mass ratio of the surfactant to the ionic polymer is as follows: (1-100) 0.1-50; the proportion of the surfactant to the solvent is as follows: 5-3000 mg of surfactant is contained in each 100mL of solvent;

preferably, the mass ratio of the surfactant to the ionic polymer is (2.5-31) to (1-7.5); the proportion of the surfactant to the solvent is as follows: each 100mL of the solvent contains 50-3000 mg of the surfactant.

Further, the surfactant is a nonionic surfactant.

Further, the nonionic surfactant is span, polysorbate, poloxamer, alkyl glucoside, vitamin E polyethylene glycol succinate, sucrose stearate or azone.

Further, the ionic polymer is selected from carboxymethyl cellulose and its salt, sodium starch glycolate, hyaluronic acid and its salt, xanthan gum, alginic acid and its salt, and polyethylene glycol diacetate PEG- (COOH)₂At least one of (1).

The invention also provides a carrier or an auxiliary material of the ophthalmic preparation, which comprises the following components: low-polymerization degree polyvidone and medium-polymerization degree polyvidone, and also contains solvent.

Further, the mass ratio of the low-polymerization degree povidone to the medium-polymerization degree povidone is as follows: (0.1-10) 1, wherein the ratio of the low-polymerization-degree povidone to the solvent is as follows: 5-3000 mg of low-polymerization-degree povidone is contained in every 100mL of the solvent;

Furthermore, the low-polymerization-degree povidone is povidone with the weight-average molecular weight of 2000-5000 Dalton, and preferably povidone PVP K12 with the weight-average molecular weight of 3500 Dalton.

Furthermore, the povidone with the medium polymerization degree is povidone with the weight-average molecular weight of 20000-60000 Dalton, and is preferably povidone PVP K30 with the weight-average molecular weight of 35000-50000 Dalton.

Further, the solvent in the carrier or the adjuvant of the ophthalmic preparation is a polar solvent.

Further, the polar solvent is water.

Furthermore, the carrier or the auxiliary material of the ophthalmic preparation also contains the following components: tackifiers and/or cosolvents.

Preferably, the tackifier is at least one of polyethylene glycol, carbomer, poloxamer, povidone, hydroxypropyl cellulose, methyl cellulose, hydroxyethyl cellulose, polyvinyl alcohol, xanthan gum, polyoxyethylene fatty alcohol, hyaluronic acid and salt thereof or hydroxypropyl methyl cellulose, and the cosolvent is propylene glycol, glycerol, liquid polyethylene glycol, hydrogenated castor oil or castor oil; the mass ratio of the tackifier to the surfactant or the tackifier to the low-polymerization-degree polyvidone is 1 (0.1-100), and the mass ratio of the cosolvent to the surfactant or the cosolvent to the low-polymerization-degree polyvidone is 1 (1-10) to 1;

more preferably, the mass ratio of the tackifier to the surfactant or the tackifier to the low-polymerization-degree povidone is 1 (0.1-30).

Further, the carrier or the auxiliary material of the ophthalmic preparation contains a nano corpuscle, wherein the nano corpuscle is spherical and has a particle size of 1-100 nm; the nanometer corpuscle is formed by self-assembling components of a carrier or an auxiliary material of the ophthalmic preparation.

Further, the particle size of the nano particles is 5 to 30 nm.

Furthermore, the carrier or the auxiliary material of the ophthalmic preparation also contains nano-spheres, wherein the nano-spheres are spherical and have the particle size of 10-2000 nm. The nanospheres are formed by self-assembly of nanospheres.

Furthermore, the particle size of the nano-spheres is 100 to 2000 nm.

The invention also provides a preparation method of the carrier or the auxiliary material of the ophthalmic preparation, which is to uniformly mix the components and the solvent into a solution and then grind or uniformly disperse the solution.

The invention also provides the use of the carrier or the auxiliary material of the ophthalmic preparation in the preparation of an eye drop administration carrier for delivering the medicament to the posterior segment of the eye.

The invention also provides an ophthalmic preparation for eye drop administration, which is a preparation consisting of the carrier or the auxiliary material of the ophthalmic preparation and the active component for treating the eye diseases.

Furthermore, the mass ratio of the surfactant to the active ingredient for treating the eye diseases in the carrier or the auxiliary material of the ophthalmic preparation is (1-30) to (1-2);

Furthermore, the carrier or the auxiliary material of the ophthalmic preparation contains a nano-corpuscle, and the active ingredient for treating the eye disease is wrapped in the nano-corpuscle.

Further, the aforementioned nanospheres are spherical and have a particle size of 1 to 100nm, preferably 5 to 30 nm.

Further, the active ingredients for treating eye diseases comprise small molecule compound drugs, or free acids, or free bases, or pharmaceutically acceptable salts thereof;

more preferably, the nucleoside antiviral drugs comprise ganciclovir, acyclovir, penciclovir, cidofovir, fosmivir and lobucavir;

the ocular hypotensive drug comprises carbonic anhydrase inhibitor, more preferably brinzolamide, acetazolamide, methazolamide;

the antibiotic medicines comprise amikacin, ceftriaxone, cefazolin, oxacillin, levofloxacin, ciprofloxacin, moxifloxacin and vancomycin;

the anti-inflammatory drugs include: oxytetracycline;

the antioxidant comprises taurine, anthocyanin and lignin;

the muscarinic receptor blocker drugs comprise atropine, scopolamine and anisodamine;

the glucocorticoid medicine comprises cortisone, prednisone, prednisolone, methylprednisolone, triamcinolone acetonide.

The invention also provides a method for preparing the ophthalmic preparation, which comprises the following steps:

(1) adding a surfactant and/or a tackifier into a solvent to prepare a solution;

(3) stirring and dispersing or homogenizing and dispersing the primary suspension obtained in the step (2) to obtain the suspension;

or comprises the following steps:

(a) adding low-polymerization-degree povidone and/or tackifier into a solvent to prepare a solution;

Further, the dispersion in the step (2) or the step (b) is at least one selected from the group consisting of mechanical stirring dispersion, magnetic stirring dispersion, vortex shaking dispersion, shear dispersion, homogeneous dispersion, grinding dispersion and ultrasonic dispersion.

The invention also provides application of the ophthalmic preparation in preparing a medicament for treating eye diseases.

Further, the above-mentioned ophthalmic preparation is an ophthalmic preparation for treating fundus diseases, and/or treating posterior segment viral infectious diseases, and/or treating posterior segment chronic inflammation, and/or reducing intraocular pressure, and/or ocular pain, and/or resisting intraocular bacterial or fungal infection, and/or an ophthalmic preparation for the prevention and treatment of juvenile myopia, pseudomyopia, and/or an ophthalmic preparation for the treatment of autoimmune diseases, and/or an ophthalmic preparation for the treatment of anterior segment diseases, and/or an ophthalmic preparation for the inhibition of tumor growth.

Furthermore, the ophthalmic preparation for treating fundus diseases comprises ophthalmic preparations for treating macular edema, inflammatory edema and inflammatory pain caused by fundus vascular diseases; more preferably, the ophthalmic preparation is used for treating central retinal vein obstructive macular edema, branch retinal vein obstructive macular edema, diabetic retinopathy, diabetic macular edema, pathologic myopia macular edema, macular edema caused by wet age-related macular degeneration, dry macular degeneration, geographic atrophy, noninfectious endophthalmitis, acute retinal necrosis, postoperative inflammatory pain and uveitis;

the ocular preparation for reducing intraocular pressure comprises an ocular preparation for treating acute and chronic glaucoma and complications thereof;

Further, the active ingredient of the above ophthalmic preparation against intraocular bacterial or fungal infections is an antibiotic;

the active ingredient of the ophthalmic preparation for preventing and treating juvenile myopia and pseudomyopia is a muscarinic receptor blocker;

the active ingredient of the ophthalmic preparation for treating autoimmune diseases is an immunosuppressant.

The nano corpuscle of the invention is: the components of the ophthalmic preparation carrier or the auxiliary material are self-assembled in a solvent to form a nano-scale spherical aggregate.

The nano-spheres of the invention are: the nanometer corpuscle is self-assembled in the solvent to form a spherical self-assembly structure.

The solvent of the invention is: a liquid capable of dissolving the components of the ophthalmic formulation vehicle or adjuvant.

The surfactant of the invention is: substances capable of significantly reducing the surface tension of liquids; the nonionic surfactant used in the present invention means a surfactant which does not dissociate in water.

The ionic polymer of the invention is: a high molecular polymer having a cation or an anion.

The low-polymerization-degree polyvidone is as follows: the polyvidone with a molecular weight of less than 10000Dalton, wherein the polyvidone with a medium polymerization degree is polyvidone with a molecular weight of more than 10000Dalton and less than 100000 Dalton.

The invention refers to the active components for treating eye diseases, which are as follows: the active substance which can be used for treating eye diseases, namely the active substance which is used as an ophthalmic medicine at present, and the action mechanism and the action target point show that the active substance can treat the eye diseases, but the active substance which is used as the ophthalmic medicine is not available at present.

The eye drop administration of the invention is as follows: a drug administration method for dripping liquid medicine into eyes belongs to the corneal drug administration route.

Because of the particularity of eyeball structure, the administration mode in the prior art can not give consideration to safe administration and effective administration, the mode that can effectively administer has the safety problem, and the mode that can safely administer can not effectively administer. For example, although intravitreal injection can be effectively administered, severe complications such as intraocular hemorrhage and pain exist, the currently most safe administration by eye drops is very safe, but the drug cannot reach the posterior segment of the eye due to the inability to permeate the anterior segment of the eye, and the effective concentration is insufficient, so that the purpose of effective treatment cannot be achieved.

Through years of accumulation, the inventor invents the eye drop administration carrier, and experiments prove that the eye drop administration carrier can carry the medicine, convey the medicine to the vitreous body to play a role, is stable, and solves the technical problem which needs to be solved but is not solved for a long time in the field of ophthalmic medicine administration.

Experiments show that the eye drop administration carrier can be used for conveying various medicaments, can achieve effective (expected) concentration on the eyeground, and does not influence the property and the effect of the active ingredients carried (wrapped) on the eyeground for treating the eye diseases. The eye drop administration carrier can be used for conveying the existing micromolecule medicines which are administered by vitreous injection, vitreous solution implantation, oral administration and systemic injection, further can overcome the problems of the existing vitreous injection, vitreous injection implantation, oral administration and systemic injection administration, solves the serious complication problems of intraocular hemorrhage, pain and the like, greatly reduces the pain of patients with fundus diseases, increases the medical compliance, improves the life quality of the patients and families, or avoids systemic toxic and side effects caused by systemic administration.

The invention can avoid the complication caused by local injection or implantation in eyes.

The preparation developed by the invention has small dosage and small toxic and side effects, and not only can be used as a therapeutic drug, but also can be used for preventing and controlling ophthalmic diseases.

The preparation of the invention can meet the requirement of long-term administration in clinic.

According to the eye drop administration treatment system, the Active Pharmaceutical Ingredient (API) can be small molecular drugs which are used clinically and have definite action mechanisms, the quality is controllable, the product is convenient to use, the compliance of patients is good, and doctors can flexibly adjust the administration scheme according to the illness state of the patients.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1 is a transmission electron micrograph (200 nm on a scale) of a sample (A) obtained in example 1; (B) transmission electron microscopy after staining with stain (1 μm scale).

FIG. 2 is a transmission electron micrograph (0.5 μm on a scale) of a sample obtained in example 3.

FIG. 3 is a transmission electron micrograph (500 nm on a scale) of a dyed sample obtained in example 6.

FIG. 4 is a transmission electron micrograph (200 nm on a scale) of a dyed sample obtained in example 13.

FIG. 5 is a transmission electron micrograph (0.5 μm on a scale) of a sample (A) obtained in example 36; (B) transmission electron microscopy after addition of stain (0.5 μm scale).

Detailed Description

The reagents or apparatus used in the present invention can be obtained commercially, used without specifying the conditions, and used according to the conventional conditions or conditions recommended by the manufacturer.

Some of the equipment is as follows:

ES225SM-DR (E) electronic analytical balance, Precisa corporation (Switzerland);

DF-101S heat collection type constant temperature heating magnetic stirrer, gu city ying yu gao ke jie (river south, china);

WH-2 micro vortex mixer, Shanghai West Analyzer Co., Ltd. (Shanghai, China);

a dispersion machine: t25 easy clean digital, IKA (Germany);

KQ-500 model ultrasonic cleaning apparatus, kunshan ultrasonic instruments ltd (kunshan, china);

JP-010T type ultrasonic cleaner, Shenzhen Jie union cleaning equipment Limited;

AH-NANO Plus high pressure homogenizer, anto nanotechnology (suzhou) limited (china);

PM-DK2 planetary ball mill, zhuo instruments & equipments (shanghai) limited (shanghai, china);

mettler Toledo FE20 pH meter, Mettler-Torlods (Switzerland);

NS-90 nanometer particle size analyzer, zhhai ohmaeke instruments ltd (zhhai, china);

agilent 1100HPLC, agilent technologies ltd (usa);

API4000 triple quadrupole mass spectrometer (Applied Biosystems, usa);

STY-1A osmometer, Tianjin, Tianda technology Co., Ltd (Tianjin, China).

The method for detecting the properties of the preparation of the present invention is as follows

The particle size detection method comprises the following steps:

1mL of the sample prepared in example or comparative example was transferred to a sample cell, the detection temperature was set at 40 ℃, and the sample cell was placed in an NS-90 nanometer particle size analyzer to start detection. The test was repeated 3 times for each sample, and the average of the 3 test results was taken as the particle size (in terms of light intensity distribution and percentage) and Polydispersity Index (PdI, Polydispersity Index) of the sample.

And (3) an osmotic pressure detection method:

the freezing point depression of the solution was measured to determine its osmolality. The operation is as follows: cleaning the probe of the STY-1A osmometer: three 100 mu L of distilled water are taken to be put into 3 sample tubes, after the instrument is preheated, the sample tubes filled with 100 mu L of distilled water are screwed on the probe of the instrument, and the cleaning is selected for 3 times, and the 'cleaning' is clicked and repeated for three times. And (3) detection: after filling sample information in the instrument information table, clicking 'test'; using a pipette, 100. mu.L of sample was pipetted into the sample tube, the instrument was gently screwed on, and the "start" test was clicked. The detection is repeated three times, and the average value of the 3 detection results is taken as the detection result.

The pH value detection method comprises the following steps:

the FE20 type acidimeters are respectively calibrated by pH buffer solutions (pH is respectively 4.00, 6.86 and 9.18), electrodes are washed by pure water, excess water is absorbed by fiber-free paper, the electrodes are immersed in a liquid sample to be detected to start measurement according to a reading key, and the data obtained after the reading is stable is the pH value of the sample.

If the pH of the detected solution is less than 5 or more than 9, the pH needs to be adjusted to 6-8 by acid or alkali, and commonly used pH adjusting agents are NaOH and HCl, phosphoric acid and phosphate (such as sodium dihydrogen phosphate and disodium hydrogen phosphate), citric acid and citrate (such as sodium citrate), boric acid and borax; and if the osmotic pressure of the obtained liquid is detected to be not equal to the isotonic, adding a proper amount of sodium chloride to ensure that the liquid is equal to or close to the isotonic.

Unless otherwise indicated, the efficacy of the drug delivered to the posterior segment of the eye was verified as follows:

test apparatus equipment: high performance liquid chromatograph, model: LC-20AD (Shimadzu, Japan); mass spectrometry: the model is as follows: API4000 triple quadrupole mass spectrometer (Applied Biosystems, usa); a chromatographic column: fortis Pace C185. mu.M, 2.1X30mm (Fortis Co., UK).

Rat: healthy adult Sprague Dawley (SD) rats were selected and divided into a test group and a control group, each group having 6 eyes, the test group was added dropwise with the ophthalmic preparation prepared in the examples of the present invention, and the control group was added dropwise with a suspension of 2mg of the drug in 5mL of physiological saline (vortexed well before use), 20. mu.L per eye. Animals were euthanized at 0.5 or 1 hour post-dose, vitreous samples were collected quickly, homogenized, and stored at-80 ℃. Homogenizing 10 μ L of vitreous body, adding 90 μ L of 95% ethanol, performing ultrasonic treatment for 2 min, and mixing for 1 min by vortex to obtain vitreous body homogenate; mu.L of the homogenate was taken, 175. mu.L of methanol was added, vortex mixed for 3min, centrifuged at 12000rpm at 4 ℃ for 10min, the supernatant was filtered through a 0.45 μm syringe filter and the filtrate was used for LC/MS/MS (positive ion mode, MRM SCAN) analysis.

New Zealand rabbits: healthy 3-4 month-old male New Zealand rabbits with the weight of 2.0-2.5 k g are selected and divided into two groups, and each group has 4 eyes. Grabbing New Zealand rabbits to an operating table, and after animals are calmed, dropping 30 mul of physiological saline to eyes (blank control) of 1 group of animals respectively; another group of animals was added 30 μ l of test substance to each eye, and after 1 hour the animals were euthanized, and the aqueous humor and vitreous humor from both eyes were collected rapidly and stored at-80 ℃.

Taking a New Zealand rabbit aqueous humor sample of 50 mu L, adding 50 mu L of 75% acetonitrile-water and 150 mu L of internal standard (midazolam 20ng/ml acetonitrile solution), carrying out vortex mixing for 10min, centrifuging at 4 ℃ at 10000rpm for 5min, and taking supernatant for LC-MS/MS analysis. After homogenizing a sample of a New Zealand rabbit vitreous body, 100 mul of the sample is taken, 100 mul of 75% acetonitrile-water and 50 mul of internal standard (50ng/ml acetonitrile solution) are added, vortex mixing is carried out for 10min, centrifugation is carried out at 10000rpm at 4 ℃ for 5min, and supernatant is taken for LC-MS/MS (positive ion mode, MRM SCAN) analysis.

Example 1: preparation of the ophthalmic preparation of the invention

Weighing 0.24g of CMC-Na (sodium carboxymethylcellulose, ionic polymer) according to the table 1, adding into a glass triangular flask containing 40mL of purified water, starting magnetic stirring for 2 hours to obtain a solution 1; respectively weighing 1.0g of polysorbate 80 (surfactant) and 0.24g of HPMC (hydroxypropyl methyl cellulose, tackifier) into a glass triangular flask containing 60mL of purified water, starting magnetic stirring, heating in water bath at about 40 ℃ for 1.5 hours to obtain a solution 2; weighing 40mg of dexamethasone and 4.0mL of PEG400 (namely 4.3 times (w/w) of the dosage of the surfactant) and putting into the solution 2, continuing heating and stirring for 30 minutes, adding the solution 1 and stirring for 30 minutes to obtain a mixed solution; dispersing the mixed solution for 5 minutes at 9500 r/m by using a dispersion machine, stopping the machine until the foam disappears, and filtering the mixture by using a Buchner funnel under reduced pressure to obtain a dispersion solution; and transferring the dispersion liquid to a high-pressure homogenizer, controlling the temperature to be 15 +/-5 ℃, homogenizing for 3 minutes under the pressure of 400Bar, then increasing the pressure to be more than 800Bar for homogenizing for 25 minutes, reducing the pressure to be 300Bar for homogenizing for 2 minutes, discharging to obtain a colorless clear solution, further reducing the pressure, filtering and sterilizing, and removing mechanical impurities to obtain the colorless clear solution after impurity removal.

pH and osmotic pressure adjustment: adding 700mg NaH₂PO₄And 400mg of Na₂HPO₄Adjusting the pH to 6.3; sodium chloride was added to adjust the osmotic pressure to 282 mOsmol/kg.

And (4) HPLC detection: a detection instrument: agilent 1100 hplc; and (3) operating software: OpenLab CDS c.01.10(201) Chemstation Edition;

chromatographic conditions are as follows: the chromatographic column is Waters Xbridge C185 μm,4.6X250 mm; the column temperature is 35 ℃, the flow rate is 1.0mL/min, the detection wavelength is 240nm, and the mobile phase: 0.1% aqueous phosphoric acid (72.0%) -acetonitrile (28.0%) isocratic elution. The sample was diluted 5 times with the mobile phase, and 10. mu.L of the diluted sample was injected into a liquid chromatograph. And (3) detection results: 96.2 percent.

Particle size 20.6nm (85.6%), PdI: 0.266. the sample is stored for 1 month at room temperature in dark, and the appearance and the content of the sample are unchanged.

The animal vitreous dexamethasone concentration 0.5 hours after eye drop is: 53.4(ng/g), RSD: 36.6 percent. The control group was dosed with a suspension of 2mg dexamethasone/5 mL saline (vortexed well before use) 20 μ L per eye, and no dexamethasone was detected in the vitreous of the animals 0.5 hours after eye drop (below the limit of detection, <1 ng/g).

Example 2:

the preparation method refers to example 1, and the raw materials and the amounts are shown in table 1. To obtain colorless clear solution after impurity removal.

And (3) pH adjustment: adjusted to pH6.5 with 0.2N NaOH or/and 0.1N HCl.

The HPLC detection method is the same as that of example 1, and the HPLC content detection result is as follows: 95.1%, particle size 12.9nm (92.1%), PdI: 0.509; the stability is good, and the appearance and the content of the product are not obviously changed after the product is placed at room temperature in a dark place for 1 month; a small amount of precipitate appeared after 2 months.

The API concentrations in rat vitreous 1 hour after eye drop were: 13.9ng/g, RSD: 17.2 percent.

Example 3:

preparation method referring to example 1, after the yellowish clear solution after impurity removal was obtained with the raw materials and the amounts shown in table 1, sodium chloride was added to adjust the osmotic pressure to: 273 mOsmol/kg.

And (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase A is 0.1% formic acid water solution, and mobile phase B is ACN. Temp.: 35 ℃, detection wavelength: 296nm, 0.8ml/min of Flowrate; gradient elution procedure: 0-2 ', 95% A-5% B, 15', 55% A-45% B,18-21 ', 35% A-65% B, 23', 95% A-5% B. And (3) detection results: 98.2 percent.

Particle sizes 13.2nm (81.2%) and 57.7nm (13.1%), PdI: 0.431; the stability is good, and the appearance and the content are not obviously changed after the glass is placed at 40 ℃ in the dark for 20 days;

the API concentrations in rat vitreous at 0.5 hours after eye drop were: 315ng/g, RSD: 29.4 percent.

API concentrations in the vitreous of new zealand rabbits 0.5 hours after eye drop were: 142ng/g, RSD: 34.3 percent.

Example 4:

preparation method referring to example 3, a yellowish clear solution after impurity removal was obtained, and the raw materials and the amounts are shown in table 1.

And (3) detection results: 16.6nm (96.2%), PdI: 0.229.

the HPLC detection method is the same as that of example 3, and the detection result is as follows: 97.5 percent; good stability, no obvious change in appearance and content after being placed at 40 ℃ in dark for 20 days.

Example 5:

preparation method referring to example 1, raw materials and amounts are shown in Table 1, and after obtaining a colorless clear solution after impurity removal, pH6.5 was adjusted with 0.1N NaOH.

And (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase A40 mM ammonium acetate aqueous solution (pH5.0), mobile phase B: MeOH, temp.: 35 ℃, detection wavelength: 233nm, 0.8ml/min Flowrate; gradient elution procedure: 0-2 ', 100% of A,20-22 ', 60% of A-40% of B and 23 ', 100% of A. And (3) detection results: 97.4 percent;

particle size 11.8nm (71.6%), PdI: 0.519. the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

Example 6:

preparation method referring to example 5, the raw materials and the amounts are shown in table 1, after obtaining a colorless clear solution after impurity removal, adjusting to ph6.5 with 1N sodium citrate solution, adding sodium chloride to adjust the osmotic pressure to: 297 mOsmol/kg.

And (4) HPLC detection: column ZORBAX 300SB-CN,2.1X150mm, 5 μm; mobile phase 40mM KH₂PO₄(pH4.5): methanol (75:25) isocratic elution, temp.: 35 ℃, detection wavelength: 233nm, 0.8ml/min Flowrate; and (3) detection results: 99.1 percent.

Particle size 21.6nm (94.4%), PdI: 0.206; the product is placed at room temperature in dark for 2 months, and the appearance and the content are not obviously changed.

The animal vitreous API concentrations at 0.5 hours after eye drop were: 39.8. + -. 16.6 ng/g.

Example 7:

preparation method referring to example 5, the raw materials and the amounts are shown in table 1, and colorless clear solution after impurity removal is obtained, and the pH detection result is as follows: 6.9, no adjustment is needed.

HPLC assay method referring to example 5, assay results: 98.6 percent; particle size 16.6nm (98%), PdI: 0.227, and the product is placed at room temperature in a dark place for 2 months without obvious changes in appearance and content.

Example 8:

HPLC assay method referring to example 5, assay results: 97.8 percent. Particle size 17.1nm (55.5%), 513 (36.3%), PdI: 0.795; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

Example 9:

the preparation method comprises the following steps: weighing 60mg of CMC-Na, adding the CMC-Na into a glass triangular flask containing 15mL of pure water, starting magnetic stirring for 2 hours to obtain a solution 1; respectively weighing 0.24g of polysorbate 80 and 0.12g of HPMC (tackifier) and adding into another glass triangular flask containing 15mL of pure water, starting magnetic stirring, heating in water bath at 40 ℃ for about 3 hours to obtain a solution 2; weighing 15mg of lipoic acid and 1mL of glycerol (equivalent to 5.25 times (w/w) of the dosage of the surfactant) and adding the lipoic acid and the glycerol into the solution 2, continuing heating and stirring for 30 minutes, adding the solution 1, and stirring for 30 minutes to obtain a mixed solution; dispersing the mixed solution for 3 minutes at the rotating speed of 11,000 revolutions by using a dispersion machine, stopping the machine until the foam of the solution disappears, transferring the solution into a high-pressure homogenizer for homogenization treatment (the conditions refer to example 1) to obtain a colorless clear solution, and then filtering and sterilizing the solution under reduced pressure and removing mechanical impurities to obtain the colorless clear solution after impurity removal;

pH and osmotic pressure adjustment methods: adjusted to ph6.3 with 0.1N sodium citrate solution, adjusted to osmotic pressure with sodium chloride: 294 mOsmol/kg;

and (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase a 0.1% phosphoric acid (ph3.0) and B methanol-acetonitrile (1: 1). Temp.: 35 ℃, detection wavelength: 215nm, 0.8ml/min Flowrate; gradient elution procedure: 0-5 ', 60% A-40% B, 28-30', 40% A-60% B; and (3) detection results: 97.4 percent. And (3) granularity detection results: 17.8nm (98.6%), PdI: 0.222; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content of the product are unchanged.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 52.6 + -17.9 ng/g.

The control group was dosed with a suspension of 2mg lipoic acid/5 mL of physiological saline (vortexed well before use) at 20 μ L per eye, and no lipoic acid was detected in the animal vitreous 0.5 hours after eye drop (below the limit of detection, <1 ng/g).

Example 10:

preparation method and pH and osmotic pressure adjustment methods referring to example 5, raw materials and amounts are shown in Table 1, and colorless clear solutions after impurity removal are obtained.

The HPLC detection method is the same as that of example 5, and the HPLC detection result is as follows: 98.4 percent

And (3) detection results: particle size 348nm (85%), PdI: 0.422.

the product is placed at room temperature in dark for 1 month, the appearance and the content of the product are unchanged, no obvious change is caused for 1 month, and a small amount of precipitate appears after 2 months.

Example 11:

preparation method and pH, osmotic pressure adjusting method referring to example 9, raw materials and dosage are shown in Table 1, wherein the dosage of cosolvent propylene glycol is 6.2 times (w/w) of surfactant.

The HPLC detection wavelength is 233nm, the other detection methods are the same as the example 9, and the detection result is as follows: 98.1 percent; particle size 25.8nm (87.4%), PdI: and (3) placing the mixture for 1 month at 0.317 and 3-8 ℃ in the dark, wherein the appearance and the content are unchanged.

Example 12:

The HPLC detection method is the same as that of example 9, and the detection result is as follows: 95.2 percent; particle size 31.5nm (82.9%), PdI: 0.347. the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are unchanged.

Example 13:

preparation method referring to example 1, raw materials and amounts are shown in table 1, and a yellowish clear solution after impurity removal is obtained.

Adjusting pH and osmotic pressure: adjusted to ph6.3 with 0.1N NaOH, and adjusted to osmotic pressure with sodium chloride: 297 mOsmol/kg;

the HPLC detection wavelength is 280nm, the other detection methods are the same as the example 1, and the detection result is as follows: 97.3 percent; particle size 16.7nm (98.1%), PdI: 0.225; the product is placed at room temperature in dark for 1 month, and the appearance and the content are unchanged.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 66.5 + -18.1 ng/g.

The control group was dosed with a suspension of 2mg doxycycline/5 mL of physiological saline (vortexed well before use) at 20 μ L per eye, and no doxycycline was detected in the vitreous of the animals 0.5 hours after the control group was dosed with eye drops (below the limit of detection, <1 ng/g).

Example 14:

preparation method and pH and osmotic pressure adjustment method referring to example 13, raw materials and amounts are shown in Table 1, and a yellowish clear solution after impurity removal is obtained.

The HPLC detection method is the same as that of example 13, and the HPLC detection method comprises the following detection results: 98.2 percent; particle size 17.2nm (97.9%), PdI: 0.208; the product is placed at room temperature in dark for 1 month, and the appearance and the content are unchanged.

Example 15:

preparation method and pH, osmotic pressure adjustment method referring to example 13, the addition of co-solvent was: 1.5mL of propylene glycol (corresponding to 10 times (w/w) of the surfactant) and the surfactant were weighed and added into medium water, magnetically stirred, and heated in a water bath to dissolve the mixture to obtain a solution 2, which was a yellowish clear solution after impurity removal, and the raw materials and the amounts thereof were as shown in Table 1.

The HPLC detection method is the same as that of example 13, and the detection result is as follows: 95.2 percent; particle size 29.7nm (89.3%), PdI: 0.382; and placing the mixture for 1 month at 3-8 ℃ in the dark, wherein floccules appear.

Example 16:

preparation method and pH and osmotic pressure adjustment method referring to example 9, raw materials and amounts are shown in Table 1.

HPLC assay method referring to example 9, assay results: 0.486mg/mL (metformin), 0.481mg/mL (lipoic acid); particle size 18.9nm +302.1nm, PdI: 0.529; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are unchanged.

The animal vitreous API concentrations at 0.5 hours after eye drop were: 86.5ng/g lipoic acid, 69.5ng/g metformin.

Example 17:

preparation method and pH and osmotic pressure adjustment method referring to example 9, raw materials and amounts are shown in Table 1.

HPLC assay method referring to example 9, assay results: 0.487mg/mL (doxycycline), 0.478mg/mL (lipoic acid); particle size 20.2nm +251.6nm, PdI: and the product is placed at 0.701 and 3-8 ℃ in the dark for 1 month, and the appearance and the content are unchanged.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 57.3ng/g lipoic acid, 68.4ng/g metformin.

Example 18:

the preparation method refers to example 1, the raw materials and the dosage are shown in table 1, after the colorless clear solution after impurity removal is obtained, sodium chloride is added to adjust the osmotic pressure to: 301 mOsmol/kg; and (3) pH detection result: 6.6 No adjustment is required.

And (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase a 0.1% phosphoric acid, mobile phase B acetonitrile, temp.: 35 ℃, detection wavelength: 260nm, 0.8ml/min Flowrate; gradient elution procedure: 0-2 ', 95% A-5% B,20-25 ', 65% A-35% B,28 ', 95% A-5% B; and (3) detection results: 98.2 percent. Particle size 20.3nm (83.6%), PdI: 0.249; the product is placed at room temperature for 1 month, and the appearance and the content of the product are unchanged.

API concentrations in the vitreous of new zealand rabbits 1 hour after eye drop were: 138ng/g, and the concentration in the aqueous humor is 681 ng/g.

Example 19:

preparation method and pH and osmotic pressure adjustment methods referring to example 1, raw materials and amounts are shown in Table 1, and a yellowish clear solution after impurity removal is obtained.

And (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase a 0.1% phosphoric acid, mobile phase B methanol (80:20) isocratic elution, temp.: 35 ℃, detection wavelength: 280nm, 0.8ml/min Flowrate; and (3) detection results: 98.4 percent. Particle size 39.7nm (95.5%), PdI: 0.318; the product is placed at room temperature in dark for 1 month, and the appearance and the content are unchanged.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 78.3 ng/g.

Example 20:

preparation method and pH and osmotic pressure adjustment referring to example 19, raw materials and amounts are shown in Table 1. A yellowish clear solution after impurity removal is obtained.

The HPLC detection method is the same as that of example 19, and the detection result is as follows: 97.8 percent; particle size: 46.2nm (95.5%), PdI: 0.343; the appearance and the content of the product are obviously unchanged after being placed at room temperature in the dark for 1 month.

Example 21:

the preparation method refers to example 1, the raw materials and the dosage are shown in table 1, a colorless clear solution after impurity removal is obtained, and sodium chloride is added to adjust the osmotic pressure to 271 mOsmol/kg; and (3) pH detection result: 6.6, no adjustment is needed.

And (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase 0.1% H₃PO₄-acetonitrile (85:15) isocratic elution, temp.: 35 ℃, detection wavelength: 217nm, 0.7ml/min Flowrate; and (3) detection results: 99.6 percent. Particle size 15.2nm (93.4%), PdI: 0.227; the product is placed at room temperature in dark for 1 month, and the appearance and the content are obviously unchanged.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 79.4 (ng/g).

Example 22:

the preparation method refers to example 21, the raw materials and the dosage are shown in table 1, the colorless clear solution after impurity removal is obtained, and sodium chloride is added to adjust the osmotic pressure to 265 mOsmol/kg; and (3) pH detection result: 6.5, no adjustment is needed.

The HPLC detection method is the same as that of example 21, and the detection result is as follows: 98.3 percent; particle size 11.9 (91.9%) nm, PdI: 0.206; the appearance and the content of the product are not obviously changed after being placed at room temperature in the dark for 1 month.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 46.2 (ng/g).

Example 23:

the preparation method refers to example 9; the raw materials and the dosage are shown in table 1, wherein the dosage of the cosolvent propylene glycol is 4.5 times (w/w) of the surfactant, the pH is 6.5, and the surfactant is nearly isotonic and does not need to be adjusted.

HPLC detection method: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; the mobile phase A is 0.1 percent of phosphoric acid, and the mobile phase B is acetonitrile (80:20), and isocratic elution is carried out; temp.: 35 ℃, detection wavelength: 306nm, 0.8ml/min Flowrate; and (3) detection results: 95.7 percent;

particle size 27.5nm (77.9%), PdI: 0.328; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 20.3. + -. 9.3 ng/g.

Example 24:

the preparation method refers to example 23, the raw materials and the dosage are shown in Table 1, wherein the dosage of the cosolvent propylene glycol is 4.5 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

The pH is 6.6, and the pH is not required to be adjusted; and (3) osmotic pressure regulation: adding sodium chloride to adjust osmotic pressure: 293 mOsmol/kg;

HPLC assay method referring to example 14, assay results: 96.1 percent;

particle size 24.5nm (85.5%), PdI: 0.253; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are not obviously changed.

Example 25:

the preparation method refers to example 23, the raw materials and the dosage are shown in Table 1, wherein the dosage of the cosolvent glycerol is 4.5 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

The pH is 6.4, and the pH is not required to be adjusted; and (3) osmotic pressure regulation: adding sodium chloride to adjust the osmotic pressure to 305 mOsmol/kg;

HPLC assay method referring to example 14, assay results: 94.7 percent;

particle size 26.2nm (75.2%), PdI: 0.325; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are not obviously changed.

Example 26:

Adjusting pH and osmotic pressure: adjusted to ph6.2 with 0.2N NaOH, and adjusted to osmotic pressure with sodium chloride: 305 mOsmol/kg;

HPLC assay method referring to example 14, assay results: 95.3 percent;

particle size 22.7nm (83.4%), PdI: 0.372; the product is placed at 3-8 ℃ in the dark for 1 month, and the appearance and the content are not obviously changed.

Example 27:

preparation method referring to example 1, the raw materials and the amounts are shown in table 1, and colorless clear solution after impurity removal is obtained, and the pH is adjusted to 6.2 by 1M sodium citrate aqueous solution, and sodium chloride is added to adjust the osmotic pressure to: 307 mOsmol/kg;

and (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase A, water: mobile phase B is methanol, and the Flowrate is 0.8 ml/min; gradient elution procedure: 0-10 ', 100% A-0% B,15 ', 55% A-45% B,18-21 ', 35% A-65% B; temp.: 30 ℃, detection wavelength: 255 nm; and (3) purity detection result: 99.2 percent; particle size 19.6nm (75.9%), PdI: 0.424; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 580 ng/g.

Example 28:

the preparation method refers to example 9, the raw materials and the dosage are shown in Table 1, wherein the dosage of the cosolvent propylene glycol is 5 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

Adjusting pH and osmotic pressure: adjusted to ph6.3 with 0.1N NaOH, and adjusted to osmotic pressure with sodium chloride: 290 mOsmol/kg;

HPLC assay method referring to example 9, assay results: 96.1 percent;

particle size 23.7nm (84.2%), PdI: 0.323; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 62.5 ng/g.

Example 29:

the preparation method refers to example 1, the raw materials and the dosage are shown in table 1, wherein the dosage of the cosolvent PEG400 is 5 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

pH and osmotic pressure adjustment: with 1M Na₂HPO₄The solution was adjusted to ph6.2 and sodium chloride was added to adjust the osmotic pressure to: 295 mOsmol/kg;

the HPLC detection method is the same as that of example 1, and the detection result is as follows: 97.3 percent; particle size 22.4nm (91.4%), PdI: 0.293; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 42.7 ng/g.

Example 30:

the preparation method refers to example 9, the raw materials and the dosage are shown in Table 1, wherein the dosage of the cosolvent propylene glycol is 4.2 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

Adjusting pH and osmotic pressure: adjusted to ph6.3 with 0.1N NaOH, and adjusted to osmotic pressure with sodium chloride: 288 mOsmol/kg;

HPLC assay method referring to example 9, assay results: 95.6 percent; particle size 24.1nm (81.5%), PdI: 0.357; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

Example 31:

the preparation method refers to example 5, the raw materials and the dosage are shown in Table 1, wherein the dosage of the cosolvent propylene glycol is 5.2 times (w/w) of the surfactant, and colorless clear solution after impurity removal is obtained.

Adjusting pH and osmotic pressure: adjusted to ph6.3 with 0.2N NaOH, and adjusted to osmotic pressure with sodium chloride: 310 mOsmol/kg;

HPLC assay method referring to example 5, assay results: 97.2 percent; particle size 27.5nm (79.6%), PdI: 0.364; standing at room temperature in dark for 1 month to obtain visible microparticles.

Example 32:

the preparation method refers to example 1, and the raw materials and the amounts are shown in table 1. Obtaining colorless clear solution after impurity removal, adjusting the pH value to 6.2 by using 1M sodium citrate solution, and adding sodium chloride to adjust the osmotic pressure to 308 mOsmol/kg;

and (4) HPLC detection: column: ZORBAX Eclipse Plus C18,4.6X100mm 3.5.5 μm; mobile phase A is 0.1% phosphoric acid water solution, and mobile phase B is methanol. Temp.: 35 ℃, detection wavelength: 280nm, 0.8ml/min Flowrate; gradient elution procedure: 0-2 ', 85% A-15% B,15-20 ', 35% A-65% B,22-25 ', 15% A-85% B; and (3) detection results: 95.3 percent; particle size 17.6nm (93.9%), PdI: 0.229; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 185.3 ng/g.

Example 33:

the preparation method refers to example 9, and the raw materials and the dosage are shown in table 1, wherein the dosage of cosolvent castor oil is 3 times of that of the surfactant.

The HPLC detection wavelength is 280nm, the other conditions are the same as example 9, and the detection result is as follows: 94.7%, particle size 19.7nm (86.4%), PdI: 0.331; the pH value is 6.8; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 37.6 (ng/g).

Example 34

Preparation method referring to example 1, raw materials and amounts are shown in table 1, and colorless clear solution after impurity removal is obtained. pH6.5 need not be adjusted;

the HPLC detection wavelength is 245nm, the other detection conditions are the same as those of the example 1, and the detection result is as follows: 98.1 percent; particle size 18.6nm (96.9%), PdI: 0.257; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 43.8 ng/g.

Example 35:

the starting materials and amounts are shown in table 1, the preparation process is referred to example 1. To obtain colorless clear solution after impurity removal.

And (3) detection results: particle size 18.6nm (96.5%), PdI: 0.218;

HPLC content detection results: 97.2 percent;

after being placed at room temperature in dark place for 1 month, the appearance and the content have no obvious change.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 5.1 ng/g.

Example 36:

the starting materials and amounts are shown in table 1, the preparation process is referred to example 1. The added mass of the cosolvent PEG400 is equal to that of the surfactant (1:1(w/w)), so that a colorless clear solution after impurity removal is obtained.

And (3) detection results: particle size 19.6nm (97.0%), PdI: 0.289; ph6.33, osmotic pressure: 275 mOsmol/kg; HPLC content detection results: N/A.

After being placed at room temperature in dark place for 1 month, the appearance and the content have no obvious change.

Comparative example 1

Preparation method and detection method referring to example 1, raw materials and amounts are shown in table 1.

And (3) detection results: particle size 10.0nm (98.8%), PdI: 0.363, stability: the precipitate is formed after the mixture is placed in a cool place for 1 month.

Comparative example 2

Preparation method and detection method referring to example 1, raw materials and amounts are shown in table 1.

And (3) detection results: particle size 196nm (61.6%), PdI: 0.828, stability: the mixture is placed in a shade place for 1 month, and flocculent precipitates are generated. Taking supernatant to detect the particle size result: 530nm (53.9%) and 225nm (46.1%), PDI: 1.00,

example 37:

weighing 0.25g of povidone (PVP) K30 according to Table 2, adding to a glass triangular flask containing 15mL of purified water, and magnetically stirring for 2 hours to obtain solution 1; respectively weighing 60mg of HPMC and 60mg of povidone (PVP) K12, adding into another glass triangular flask containing 15mL of purified water, starting magnetic stirring, heating in water bath at 40 ℃ for 2 hours, and obtaining a solution 2; weighing 36mg of moxifloxacin hydrochloride, adding the moxifloxacin hydrochloride into the solution 2, continuing to heat and stir for 30 minutes, adding the solution 1, and stirring for 30 minutes to obtain a mixed solution; the bacterial and mechanical impurities were removed by the same dispersion, high-pressure homogenization and membrane filtration operations as in example 3, obtaining a yellowish clear solution after impurity removal, adjusting the osmotic pressure by adding sodium chloride to: 285 mOsmol/kg;

and (3) detection results: particle size 14.0nm (54.2%), 222nm (31.9%) and 10.1nm (13.1%), PdI: 0.564; HPLC content detection results: 96.4 percent

Stability: the appearance and the content of the product are not obviously changed after being placed at 40 ℃ for 20 days. The particle size is increased, and the detection result is as follows: 242nm (77.9%) and 18.1nm (12.4%).

Example 38:

the preparation method refers to example 1, the raw materials and the dosage are shown in table 2, wherein the cosolvent PEG400 is 5 times (w/w) of the low-polymerization degree povidone. Obtaining colorless clear solution after impurity removal, and the pH value is 6.5 without adjustment;

the HPLC detection method is the same as that of example 1, and the detection result is as follows: 98.1 percent;

particle size 15.1nm (87.1%) and 3.1nm (11.0%), PdI: 0.288; the product is placed at room temperature in dark for 1 month, and the appearance and the content are not obviously changed.

The API concentrations in rat vitreous at 0.5 hours after eye drop were: 5.1 ng/g.

Example 39:

the starting materials and amounts are shown in Table 2, the preparation and HPLC detection methods refer to example 21. And (3) pH detection result: 6.68, no adjustment is required.

And (3) HPLC detection result: 99.3 percent; particle sizes 11.5nm (62.9%) and 77.8nm (23.6%), PdI: 0.362; the appearance and the content of the product are not changed after the product is placed at room temperature in dark for 1 month.

Example 40:

the starting materials and amounts are shown in table 2, the preparation process is referred to example 1. To obtain colorless clear solution after impurity removal.

HPLC assay method referring to example 1, assay results: 98.5 percent; particle size 125.6nm (63.5%), 13.6nm (33.1%), PdI: 0.255; after standing at room temperature in the dark for 1 month, a pale emulsion was formed with a white precipitate.

Comparative example 3

The raw materials and the amounts thereof are shown in table 2, and the preparation method refers to example 1, and replaces povidone with ionic polymer CMC-Na to obtain light white emulsion.

And (3) detection results: particle size 1299nm, PdI: 0.175, standing overnight, a white precipitate formed.

The result of the measurement of the API concentration in the vitreous body of a rat after the eye drop administration shows that the ophthalmic medicine can carry active ingredients for treating eye diseases to pass through the barrier of an eyeball structure, can deliver effective dose of the medicine to the vitreous body in a conjunctival sac administration (eye drop administration) mode, avoids invasive administration modes such as vitreous body injection and the like, greatly reduces the total medicine amount, reduces the absorption of the medicine in the whole body and avoids generating toxic and side effects.

TABLE 1

TABLE 2

The beneficial effects of the ophthalmic preparation carrier or the auxiliary material are proved by the following experimental examples.

Experimental example 1 Transmission Electron microscopy results of the inventive Carrier

Transmission electron microscope (JEM-2100Plus, JEOL Ltd., Japan)

Sucking 1 drop of the liquid sample prepared in the embodiment 3-1 into a copper sample net, standing for 5 minutes, sucking off redundant liquid samples, naturally drying, and placing in an electron microscope sample chamber for detection; dyeing a sample: sucking 1 drop of liquid sample into a copper sample net, adding 1 drop of 2% phosphomolybdic acid after removing redundant sample on the sample net, standing for 5 minutes, sucking redundant liquid, naturally draining, and placing the sample net on an electron microscope for detection. The result is shown in fig. 1, it can be seen that the drug-loaded carrier prepared by the invention forms a spherical structure (nano-particles, fig. 1A) with a particle size of 1-100 nm in a solvent, and the nano-particles can be further self-assembled into spheres (nano-spheres, fig. 1B) with a particle size of 10-2000 nm.

Experimental example 2 particle size, content and stability test

1. Experimental methods

1mL of the samples prepared in examples and comparative examples was transferred to a sample cell, the detection temperature was set to 40 ℃, and the sample cell was placed in an NS-90 nanometer particle size analyzer to start detection. The test was repeated 3 times for each sample, and the average of the 3 test results was taken as the particle size (in terms of light intensity distribution and percentage) and Polydispersity Index (PdI, Polydispersity Index) of the sample. And (5) storing in dark after detection, observing appearance change and detecting the particle size again.

The HPLC content of the ophthalmic preparation sample prepared by the invention is detected by an Agilent 1100 high performance liquid chromatograph.

2. Results of the experiment

See tables 4, 5:

table 4:

table 5:

the results show that the carrier or the auxiliary material prepared by the invention can successfully entrap various types of ophthalmic medicines to prepare the ophthalmic preparation, and the preparation has small particle size, high medicine content detected by HPLC, and stable form and content after long-time storage; the carrier or the auxiliary material prepared by the invention has high drug encapsulation efficiency and good stability for eyes. The preparation prepared by using the comparative example which is different from the auxiliary raw material of the invention has poor stability, and precipitation or deterioration can occur in a short time.

Experimental example 3 antiviral experiment of ophthalmic preparation prepared with vehicle or adjuvant of the present invention

1. Subject: ganciclovir eye drop delivery systems prepared in examples 3-27

2. The experimental method comprises the following steps:

the ganciclovir eye drop delivery system prepared in the examples was tested for in vitro anti-human cytomegalovirus.

Human embryonic lung fibroblasts (MRC-5) were infected with the human cytomegalovirus strain HCMV-AD169 and the antiviral activity of the ganciclovir test article was examined in half the effective amount (EC 50). The test was divided into 4 groups, and the eye drop administration system test group prepared in this example, ganciclovir bulk drug test group, negative control group (MRC-5 cells), and positive control group (MRC-5 cells infected with HCMV-AD 169). Two test group samples were diluted with culture medium to 6 dilution concentrations: 500. 250, 125, 62.5, 31.25, 15.63 μ g/mL, 4 duplicate wells per dilution, with cytopathic effect (CPE) as an indicator: 0 is acellular lesion, 1 is not more than 25% of cytopathic effect, 2 is 25-50% of cytopathic effect, 3 is 50-75% of cytopathic effect, and 4 is 75-100% of cytopathic effect. Half the effective dose (EC50) was calculated using the Reed-Muench method: EC50 ═ 10 n; n ═ logarithm of the [ (effective rate > 50% dilution) + (percentage of effective rate > 50% — 50%)/(percentage of effective rate > 50% — percentage of effective rate < 50%) × (logarithm of dilution factor).

The assay procedure was as follows, MRC-5 cells were adjusted to approximately 1.5X 105/mL, added to a 96-well plate, 100. mu.L of culture medium was added to each well, and the mixture was placed at 37 ℃ with 5% CO₂Culturing in a cell culture box, removing supernatant after cells adhere to a monolayer, adding 100 mu L of supernatant into each hole according to the concentration 100 times of half of the amount of virus-infected cells (TC50) obtained by pre-test except for a control group, culturing for 2 hours, removing supernatant, adding 100 mu L of a series of test samples, continuously culturing, observing CPE in each hole, when the CPE of the positive control group reaches more than 90%,CPE was recorded for each well and half the effective dose was calculated according to the Reed-Muench method (EC 50).

3. The experimental results are as follows: as shown in table 6

TABLE 6 results of HCMV-AD169 virus inhibition test using samples

And (4) conclusion: the ganciclovir eye drop delivery system prepared in example 27 has no difference in the in vitro virus inhibition effect with the original drug ganciclovir.

The above results indicate that the vehicle or adjuvant of the ophthalmic preparation of the present invention, as a pharmaceutical vehicle for eye drop administration, does not affect the efficacy of the active ingredient for treating eye diseases carried (encapsulated) thereon.

Experimental example 4 in vitro bacteriostasis experiment of ophthalmic preparation prepared from the vehicle or adjuvant of the present invention

1. Subject: the moxifloxacin eye drop delivery system prepared in examples 3-3.

1.1 Experimental methods:

an in-vitro bacteriostatic test (MIC) is carried out on the moxifloxacin eye drop administration system prepared in the embodiment, a moxifloxacin raw material drug is taken as a reference substance, the moxifloxacin eye drop administration system prepared in the embodiment 6 is taken as a test product, and a drug sensitive plate is manufactured, wherein the moxifloxacin raw material drug (the reference substance) and a receiving reagent with 7-8 times of dilution concentration are contained on the drug sensitive plate. Inoculating and picking the strains (25 strains) which are incubated for 18-24 h, and preparing the strains into bacterial suspension with the turbidity of 0.5 McLeod in sterile physiological saline; adding into liquid drug sensitive test culture medium at a ratio of 1:200, mixing well, adding 100 μ L diluted bacteria solution into each hole; incubating for 16-20 h at 35 +/-2 ℃; the growth condition of bacteria in each hole is judged through turbidity, and the lowest concentration of the medicine capable of inhibiting the growth of the bacteria is the MIC of the medicine when the observation is carried out from low concentration to high concentration.

1.2 Experimental results: as shown in Table 7

TABLE 7 Moxifloxacin eye drop administration system susceptibility reagent (trace broth dilution method) test results

And (4) conclusion: the prepared moxifloxacin eye drop administration system prepared in the example 3 has no significant difference in bacteriostasis compared with a control.

The above results indicate that the carrier or adjuvant of the ophthalmic preparation of the present invention, as an eye drop administration carrier, does not affect the properties and effects of the active ingredient for treating ocular diseases carried (encapsulated) thereon.

In conclusion, the invention provides a carrier or an auxiliary material of an ophthalmic preparation and application thereof. The carrier or the auxiliary material of the ophthalmic preparation does not influence the property and the effect of an active ingredient carried (wrapped) by the carrier or the auxiliary material for the ophthalmic preparation as an ophthalmic administration carrier, can wrap the medicament to pass through the anterior segment of the eye and efficiently convey the medicament to the posterior segment of the eye to play a therapeutic role, realizes the aim of treating fundus diseases by the ophthalmic administration, solves the technical problems which need to be solved for a long time but are not solved in the field of ophthalmic preparation administration, and has extremely excellent clinical use value and very positive social significance.

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