Phospholipid nanoparticle indometacin applicable in ophthalmology

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to ophthalmology. A pharmaceutical composition is applicable in ophthalmology and contains the non-steroid anti-inflammatory drug indometacin and a phospholipid agent of the nanoparticle size 10-30 nm herbal phosphatidylcholine, maltose in the following proportions, wt %: phosphatidylcholine 20-43, maltose 55-78, indometacin 2-8. The indometacin and soya phospholipid composition represents a lyophilised powder.

EFFECT: invention provides prolonged storage stability of the composition.

2 dwg, 1 ex


The invention relates to medicine and pharmacology, and relates to stable storage and effective for its specific action of drug composition in the form of nanoparticles on the basis of vegetable phospholipids, including anti-inflammatory drug indomethacin.

Indomethacin is in a group of non-steroidal anti-inflammatory drugs and also has analgesic and antipyretic action.

The mechanism of action of indomethacin is associated mainly with the inhibition of the biosynthesis of a number of proinflammatory factors, including some of the prostaglandins that cause the inflammation to the development of pain, fever and increased tissue permeability.

Indometacin is widely used in acute and chronic arthritis of various origins, with spondyloarthropathies, arthritis, extra-articular rheumatic diseases of soft tissues (bursitis, myositis), as well as swelling and inflammation after injury or surgery (General surgical, gynecological, dental, and other). Produced indomethacin in the form of tablets, solutions for intravenous or intramuscular, rectal suppositories and forms for topical use (cream, gel), used for local anti-inflammatory effect.

In addition, indomethacin is available in powder form d is I the preparation of eye drops (0.1%)and also providing anti-inflammatory action.

The criteria for assessing the effectiveness of the new created form in vivo is or intraocular accumulation of drugs, or, in some cases, measurement of intraocular pressure.

To improve the bioavailability of ophthalmic drugs are used in various pharmaceutical approaches, such as using microemulsions, nanosuspension, nanoparticles, liposomes, dendrimers, cyclodextrins.

It was shown that the ability of the nanoparticles from polycaprolactone to interact with the mucous membrane of the eye, which stimulated a series of studies in this area, particularly in relation to anti-inflammatory drugs. Thus, it was shown that the ability of nanoparticles from chitosan containing14-indometacin or cyclosporin a, to transport the drug in the cornea and aqueous moisture, which explained the improved interaction and penetration of particles in the epithelium of the corneal layer.

For nanoparticles used natural polymers (albumin, chitosan, gelatin, starch derivatives or gelatin) or biodegradable polymers (polyalkyl-cyanoacrylate, polyacrylates). For example, the inclusion of timolol in particles based on the derived gelatin increased its bioavailability in the aqueous moisture in 5.6 times.

The inclusion of timolol-maleate in gel-obrisuyu the system, based on pluronic F-127 (polyoxyethylene+polyoxypropylene) hiked its ocular bioavailability when administered to rabbits - up to 25% compared with 2.5% for the free drugs.

There are reports on the inclusion of the drug in lipid nanoform and other ophthalmic drugs. Investigates the possibility of applying for these purposes, liposomes, particularly those with a positive charge that facilitates interaction with the cornea. For example, incorporation into liposomes of penicillin G was increased in 4 times its transport through the stratum corneum on the isolated rabbit cornea.

For delivery of glucocorticoids are used nanosuspension obtained by homogenization under high pressure. The biggest effect of this form when administered to rabbits was achieved by reducing the particle size.

Developed delivery of dexamethasone through polyoxometallates gel, using it in combination with contact lenses.

In recent years, data on the use of eye drops and gels lipid forms - solid lipid nanoparticles (SLN) and the so-called nanostructured lipid carriers (NLC), obtained by mixing the solid and liquid lipids, with specially developed relationships and technological conditions. Use combinations of such particles with non-ionic surfactants, called the a number of authors as "Nozomi" or "desmosome". Lipid nanoform also help solubilize lipophilic drugs. For example, solid lipid nanoparticles used to download tobramycin. For eye drops drugs such particles include medication is prepared in the form of nanoemulsions. When you enable indometacin in nano-emulsion was observed a greater penetration into the cornea as compared to its commercial form, especially for positively charged particles.

Phospholipid nanoparticles - lyotropic liquid crystals, chemical structure which allows to use them as carriers for both soluble and insoluble in biological fluids drugs. Injection drug compounds in phospholipid nanoparticles allows to obtain new forms of drugs with high efficacy, bioavailability and reduced side effects.

The effectiveness of the compositions of medicines on the basis of phospholipid nanoparticles is mainly determined by their size is less than 100 nm, which provides more "working surface" particles and increases their capacity for injection of drugs. The increase in the effective surface area in combination with nano creates optimal conditions for interaction of such particles to etkey. Compared to liposomes with a diameter of 250-400 nm, with an average cell size of 1-5 microns can be a single number of points of contact with the cage. For nanoparticles with a diameter of 10-30 nm this value is significantly higher, which increases the probability of their endocytosis. In addition, for such particles, there is a unique possibility of implementation in the area of intercellular contacts, the width of which in some areas can reach up to 50 nm. Thus, there is a possibility of delivery of biologically active substances inaccessible to other medicines in the areas affected tissue.

Known phospholipid transport nanosystem with a particle size of 30 nm and the possibility of inclusion of medicinal compounds with different chemical structures (RF Patent 2373924). We developed several new pharmaceutical compositions for oral and injectable forms and in vivo proved their high bioavailability and specific therapeutic activity compared to the original substances.

Thus, it seems reasonable to use a known drug substances that are embedded in the phospholipid nanoparticles smaller than 30 nm in addressing the controlled delivery of drugs for use in ophthalmology. Thanks to the development on which technology modern approaches allow to obtain phospholipid nanoparticles such a small size. One of the factors that determine the growing interest in this direction in the pharmaceutical industry - the ability to obtain stable nanoparticles with incorporated drug substances, due to which the drug appears a number of additional positive properties: the effect of passive targeting, improved bioavailability, resistance to premature degradation; the ability to penetrate the cell membrane, etc.

The purpose of the invention to provide pharmaceutical compositions on the basis of indometacin included in phospholipid nanoparticles for use in ophthalmology.

Composition one vial:

Phospholipid500 mg
Indometacin25 mg
Maltose1975 mg
Water for injection10 ml

A method of obtaining a pharmaceutical composition characterized by the following


Example 1.

A. 25 mg of indomethacin dissolved in 50 ml of rubbing alcohol when heated. To the obtained solution under vigorous stirring was added 250 mg of phospholipid and cook until complete dissolution.

Obtained races the thief is transferred into a round bottom flask with a volume of 0.5 l and evaporated on a rotary evaporator while maintaining the temperature of the water bath and vacuum level, do not allow intense boiling. After evaporation of the obtained film are harvested from the flask and weighed.

B. a portion of the maltose 500 mg dissolved in 150 ml of deionized water to the resulting solution was added to the film obtained in step a and 250 mg of phospholipid. Conduct initial homogenization of the mixture and bring it amount to 7.4 ml.

The mixture obtained at the stage of primary homogenization, is subjected to a homogenization pressure to achieve a narrow distribution of particle sizes (IP of 0.2-0.3).

C. a portion of the maltose 1475 mg dissolved in 1.7 ml of water.

, The Solutions from steps B and filtered through 0.22 μm filters and then mixed by gradual infusion of a solution of maltose in solution of phospholipid nanoparticles. The resulting solution was poured into bottles with a volume of 10 ml and dried in lyophilized.

To study the pharmacokinetics of phospholipid pharmaceutical composition in vivo as experimental animals used rats of Wistar breed contained in the standard conditions and on a standard diet, weighing 300-400 g

Study of the drug composition in experiments in vitro and in vivo

The study of the bioavailability of the pharmaceutical composition on the basis of phospholipid nanoparticles was performed in comparison with the free drug substance indomethacin. Introduction of medicines made the conduct by instillation of 25 µl sample of the drug.

Scheme of the experiment.

1. In the right eye was buried pharmaceutical composition obtained by the method described in example 1 (NF-Indomethacin).

2. In the left eye control.

Preparation of control solution.

Sample indometacin 0.025 g suspended in 10 ml of water in ultrasonic bath for 40 minutes to form a thin slurry. To 10 ml of the resulting suspension was added 1 g of maltose, mix thoroughly on a magnetic stirrer.

3. After 20 minutes taking my eyes. To do this, make a circular incision of the conjunctiva, conjunctival sephirot from the sclera, place a clamp on the optic nerve, the Central artery and vein of the retina, eye lift, scissor cross the optic nerve and blood vessels.

4. After decapitate his eyes crushed. The hanging fabric add methanol per 100 mg tissue 900 ál of solvent and homogenized until smooth. Further to 180 μl of homogenate added 20 μl of a solution of diclofenac sodium in methanol with a concentration of 5000 ng/ml and intensively stirred. The homogenate was centrifuged for 7 minutes at 13000 rpm, the Supernatant is taken and used for analysis. The samples analyzed by HPLC/MS for the determination of the drug substance.

Figure 1 shows that after 20 minutes after injection of drugs (indomethacin inpart of phospholipid nanoparticles - NF indometacin free indometacin) contents NF indometacin in tissue homogenate eyes was more than 7 times higher.

To study the specific activity of the developed medicinal composition was chosen to model non-immunogenic uveitis. As the experimental animals were taken healthy albino rabbits males, weighing 1.5-2 kg

Study the specific activity of medicinal compositions on the basis of phospholipid nanoparticles (provisional title NF-Indomethacin) was performed with the comparator drug - drug substance indometacin (Indomethacin).

For the production of this model rabbits were injected subcutaneously with 5 ml of bovine serum albumin, and then, on the 9th day, was injected 10 μl of bovine serum albumin (25 mg/ml) intravitreal.

Instillation of study drug and the comparator drug was made in the amount of 100 µl four times with intervals of 1, 5 hours, within 12 days.

Clinical evaluation of the inflammatory process in the eyes of rabbits was performed visually. Clinical manifestations of inflammation were recorded on a 1-10 day (after injection of bovine serum albumin) in conventional points for the following signs: conjunctival hyperemia, swelling of the eyelids, the amount of hypopyon (Fig.2).

Figure 2 shows that in the treatment of drug compositions NF Indometacin reduction of signs is asplenia took place on 5-6 day after the introduction and complete recovery of the animals were observed for 8 days, while during treatment with Indomethacin reduced signs of inflammation were observed at 9 days and full recovery for 12 hours.

Pharmaceutical composition for use in ophthalmology on the basis of indometacin and phospholipid nanoparticles with a size of 10-30 nm, including 78-95%phosphatidylcholine vegetable origin, maltose and indomethacin in the following ratio, wt.%:



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