RU2469706C2 - Pharmaceutical composition for transdermal application for increase of drug activity and decrease of side effects - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: phospholipid emulsion contains lecithin with the phosphatidylcholine content min. 60%, ethanol, water and diclofenac sodium in the amounts specified in the patent claim. The phospholipid emulsion represents liposomes (nanosomes) of a mean diameter of 20 to 500 nm. The composition is used for the liposomal introduction of biologically active substances and/or drugs into a human body. The emulsion nanosomes may additionally contain one or more substances specified in a group: carotinoids, flavonoids, vitamins, resveratrol, vitamin-like substances, cartilage protectors, amino acids, preserving agents.

EFFECT: emulsion provides selectivity of the effect of the active compound on body tissues, enables adjusting skin penetration rate of the active substance, dosing the active ingredients, increasing bioavailability of the active substances, reducing their negative effect of the body.

13 cl, 7 dwg, 1 tbl, 11 ex

Description

The invention relates to the chemical and pharmaceutical industry, in particular, to a composition that allows for the inclusion in the composition of liposomes (nanosomes) of drug substances and / or biologically active substances intended for transdermal use. A composition is proposed, which is liposomes (nanosomes) with a double layer of membranes, the average diameter of liposomes (nanosomes) is from 20 to 500 nm in the following optimal ratio of liposomes (nanosomes). The composition is intended for liposomal administration of biologically active substances and / or drugs into the human body.

Having appeared at the dawn of civilization, dosage forms have passed a long and complex path of evolution, during which some disappeared or changed, others appeared. An optimally selected dosage form for a particular drug substance allows you to maximize the effect of drugs with minimal side effects, change the nature of the action of the substance - accelerate or prolong it, reduce the allergenic effect, and if necessary, improve organoleptic characteristics.

One of the modern directions in the modification of traditional dosage forms is the use of directional transport for the delivery of medicinal and biologically active substances. Carriers used for the targeted transport of biologically active compounds are divided into 3 groups:

1st generation - microcapsules, microspheres;

2nd generation - passive colloidal carriers: liposomes, nanocapsules, etc .;

3rd generation - colloidal carriers with monoclonal antibodies as a vector, with a molecular substrate.

Dosage forms with carriers of the 1st generation are introduced into the vascular bed near the target. Biodegradable microspheres or microcapsules are used for prolonged release of proteins during injection of peptide hormones, small doses of steroids used as contraceptives, for prolonging the action of drug antagonists, and are used in oncology.

Carriers of the 2nd generation (sizes less than 1 micron) - liposomes - differ in numerous advantages, they can include various medicinal substances. Liposomal dosage forms can be controlled by acting externally locally in the area of the pathological process site, providing targeted delivery. Liposomes are administered in various ways: they are used to treat intracellular infections, liver, ophthalmology and externally.

Today, the transdermal route of administration of the drug is important for the treatment of skin diseases, soft tissue diseases, varicose veins, the musculoskeletal system of a person, etc. By improving this route of delivery of active substances into the human body, a transdermal therapeutic delivery system (TTC) has been developed.

For example, a transdermal delivery system is known for topical application of one or more active agents contained in one and / or several layers of a polymer and / or adhesive carrier, directly adjacent to a drug-free polymer and / or adhesive coating, which is applied either on a carrier base , or on the detachable laying of the transdermal system. This device for transdermal delivery is made by optimizing the loading of the drug while ensuring the desired adhesion to the skin or mucous membrane, as well as providing the ability to modulate the delivery of the drug and its profile (Patent RU 2356580).

The use of liposomes as a delivery route for biologically active substances has gained practical application in the cosmetic field.

Known antimicrobial liposome composition and shower gel, including liposome composition - a detergent for skin care. The antimicrobial liposome composition as a biologically active component contains dioxidine, as a component forming liposomes-phospholipids and / or cyclodextrins, a preservative selected from a number of parabens and water (Patent RU 2371166).

As a delivery method, a liposome transport route for drugs is known for oral and aerosol routes of administration. The leading positions in research and development of liposomal forms of drug administration are held by three American companies: The Liposome Company (TLC), Liposome Technology Inc. (LTI), Vestar. Thanks to their studies, liposomal amphotericin B has already been introduced to the market for the treatment of systemic mycoses, antitumor liposome preparations daunorubicin, doxorubicin, cisplatin. The final stages of clinical trials are preparations such as liposomal vaccines against influenza and melanoma, the antidiabetic complex of insulin-liposomes, antiviral nucleosides for the treatment of AIDS, a series of liposomal bronchodilator drugs, etc.

A high-dose liposome aerosol composition is described, containing 12-30 mg / ml cyclosporin A or budesonide and 130-375 mg / ml phospholipid in ultrapure water, located in a nebulizer tank, where the particle size of these liposomes is in the range from about 1 μm to about 3, 0 μm (Patent RU 2174390).

Known pharmaceutical composition for the prevention and / or treatment of cancer, to increase the activity of anti-cancer drugs and reduce the side effects of anti-cancer drugs, containing nanoparticles with high affinity for degenerate cells. This composition is proposed for oral use (patent application RU 2008110324/15 from 08.21.2006).

Known dental transdermal elixir containing liposomes, which does not have a general toxic, irritating and allergenic effect, has a uniform consistency, is evenly distributed when rinsing on the oral mucosa, has anti-inflammatory and keratoplastic effects (Patent RU 2327445).

Liposomes, as an effective mode of transport of biologically active substances of plant and natural origin, have been recognized and used especially in the cosmetic industry.

A gel is known for the treatment, prevention of the formation of keloid and hypertrophic skin scars, containing a biologically active additive in the form of a suspension of liposomes from phospholipids and lipids of animal and vegetable origin with a chloroform extract of medicinal herbs included in the gel-forming structure (Patent RU 2008110804).

A cosmetic product is described for aging skin, as well as an anti-burn and anti-herpetic drug, which uses liposomes for targeted transport of drugs (DNA molecules) (Patent RU 95102712).

Known cosmetic product for the care of problem skin, which as a liposome contains a suspension of liposomes of plant and animal origin (Patent RU 2241440).

A phospholipid emulsion comprising dihydroquercetin and a method for its preparation, in particular a liposomal biologically active form of dihydroquercetin and a method for its preparation are described. The phospholipid emulsion contains dihydroquercetin, one of the membrane-forming substances: phospholipid, glycine and an aqueous solution of ethyl alcohol with an alcohol content of 0.05 to 20% (Patent RU 2369383).

This composition by technical nature and subject is the closest to the proposed invention.

The disadvantages of this tool include the following:

- the composition is not intended to be included in the composition of medicinal substances (is a carrier of only plant biologically active substances);

- the area of practical use of the composition is limited to dietary supplements for food and cosmetics;

- insufficient bioavailability through the skin (the structure of liposomes is single-layered, in contrast to skin membranes having a double layer);

- the composition does not provide for the selectivity of the effects of the active compounds on body tissue;

- the composition does not allow you to adjust the rate of penetration of the active substance through the skin;

- the composition does not provide for the dosage of the active components;

The aim of the invention is to create a composition, the development of which takes into account all the disadvantages of the previously described products and which is intended for use in pharmaceutical practice for transdermal use. A composition is proposed, which is liposomes (nanosomes) with a double layer of membranes, the average diameter of liposomes (nanosomes) is from 20 to 500 nm, with the following optimally selected ratio of liposome components, in wt.% (Per 100 wt.%):

Ethanol 1.0-30.0 Phospholipids (Lecithin) 1.0-30.0 (with a phosphatidylcholine content of at least 60%) Water up to 100.00

This composition includes a pharmacologically active agent in an amount of from 0.001 to 30%, depending on the recommended dosage and concentration planned to be obtained in the final dosage form. The composition is designed for direct human consumption and / or for the preparation of preparations for external or internal use.

As the active component may be a biologically active component of plant origin and / or drug (s) substance (s).

The composition is designed for direct human consumption and / or for the preparation of preparations for external or internal use, for example:

The composition may be contained in the composition of the emulsion (based on 100 wt.%):

Ethanol 1.0-30.0 Phospholipids (Lecithin) 1.0-30.0 Fatty Acid Triglycerides 1.0-24.0 Coenzyme Q10 (ubiquinone) 0.5-15.0 Water 1.0-96.5

As the active component may be a biologically active component of plant origin and / or drug (s) substance (s).

The composition with the medicinal substance may be contained in the composition of the ointment base (based on 100 wt.%):

Liposomes containing diclofenac sodium 1,0-75,0 (including: ethanol 1.0-30.0%, phospholipids (lecithin) 1.0-30.0%, diclofenac sodium 0.001-30.0%, water up to 100.0%) Polyethylene Oxide-400 5.0-50.0 Polyethylene Oxide-4000 5.0-50.0 1,2-propylene glycol 5.0-50.0 Nipagin 0.01-1.0 Nipazole 0.01-1.0

A composition with a biologically active component may be contained in the gel (based on 100 wt.%):

Resveratrol Liposomes 1,0-90,0 (including: ethanol 1.0-30.0%, phospholipids (lecithin) 1.0-30.0%, resveratrol 0.001-30.0%, water up to 100.0%) Nipagin 0.1 Nipazole 0.1 Gel crosslinked polyethylene oxide-1500 up to 100.0

Nanosomes of the composition may contain one or more pharmaceutically and pharmacologically compatible active substances selected from the groups: carotenoids, flavonoids, vitamins, polyphenols, phytoalexins (preferably resveratrol), vitamin-like substances, essential substances, amino acids, proteins, minerals, angiotensin-converting enzyme inhibitors; α-adrenergic agonists; β-adrenergic agonists; α-adrenergic blockers; β-adrenergic blockers (beta blockers); alcohol inhibitors; aldose reductase inhibitors; aldosterone antagonists; amino acids; anabolics; analgesics (both narcotic and non-narcotic); anesthetics; anorexics; antacids; anthelmintic drugs; anti-acne agents; antiallergic drugs; antiandrogens; antianginal agents; sedatives; antiarrhythmics; anti-asthma drugs; antibacterial agents and antibiotics; drugs against alopecia and baldness, anti-amoebic drugs; antibodies; anticholinergics; anticoagulants and blood thinners; anticolytic drugs; anticonvulsants; anti-cystic drugs; antidepressants; antidiabetic agents; antidiarrheal agents; antidiuretics; antidotes; antiemetics; antiestrogens; antiflating agents; antifungal agents; antigens; anti-glaucoma agents; antihistamines; antihyperactive drugs; antihyperlipoproteinemic agents; antihypertensive agents; anti-hyperthyroid agents; antihypertensive drugs; antihypothyroid drugs; anti-infectious agents; anti-inflammatory (both steroidal and non-steroidal) drugs; antimalarial agents; anti-migraine agents; antineoplastics; anti-obesity drugs; antiparkinsonian agents and antidiscinetics; antipneumonic agents; antiprotozoal agents; antipruritic drugs; antipsoriatics; antipsychotics; antipyretics; anti-rheumatic drugs; antisecretory agents; anti-shock medicines; antispasmodics; antithrombotic agents; anti-cancer agents; antitussive drugs; antiulcer drugs; antiviral agents; anxiolytics; bactericidal agents; bone compactors; bronchodilators; calcium channel blockers; carbonic anhydrase inhibitors; cardiotonics and pacemakers; chemotherapeutic agents; choleretic drugs; cholinergic agents; agents for treating chronic fatigue syndrome; CNS stimulants; coagulants; contraceptives; agents for the treatment of cystic fibrosis; decongestants; diuretics; dopamine receptor agonists; dopamine receptor antagonists; enzymes; estrogen; expectorant; agents for treating gastric hyperactivity; glucocorticoids; hemostatic agents; coenzyme A reductase inhibitors of human menopausal gonadotropin; hormones; sleeping pills; immunomodulators; immunosuppressants; laxatives; agents for treating diseases of the oral cavity and periodontal diseases; myotics; monoamine oxidase inhibitors; mucolytic agents; agents for treating multiple sclerosis; muscle relaxants; mydriatics; narcotic antagonists; NMDA receptor antagonists; oligonucleotides; ophthalmic preparations; accelerating agents; peptides, polypeptides and proteins; polysaccharides; progestogens; prostaglandins; protease inhibitors; respiratory stimulants; sedatives; serotonin uptake inhibitors; sex hormones, including androgens; drugs used to combat smoking; smooth muscle relaxants; smooth muscle stimulants; thrombolytics; tranquilizers; urine acidifiers; medicines for the treatment of urinary incontinence; vasodilators; vasoprotectants; chondroprotectors (chondroitin, glucosamine and their salts, hyaluronic acid and its salts), and their combinations.

The composition may further comprise at least one vitamin selected from the group consisting of vitamin E, vitamin C, vitamin D, folic acid, vitamin B group, vitamin A, biotin, vitamin PP, pantothenic acid or its salts, vitamin K, and / or vitamin-like substances.

Highly purified lecithin with a phosphatidylcholine content of at least 60% is used as the starting material for nanosomes, since this compound, being a component of the skin cell membrane, gives the resulting nanosomes increased flexibility, mobility, penetration, and stability, participating in the formation of a stable liposome system.

The inventive preparation differs from the prototype in qualitative and quantitative composition, and also has an original production scheme and a wider field of practical use.

This goal is achieved by obtaining liposomes (nanosomes) from highly purified lecithin containing nanoparticles of pharmacologically active substances (a). To obtain a phospholipid emulsion, in terms of phosphatidylcholine, a solution of lecithin in ethanol and a dispersion medium, including deionized water, are prepared, the dispersion medium is untwisted until a stable rotation funnel is formed in it. Pharmacologically active components are added according to the formulation, while continuing mixing. The technical result is liposomes (nanosomes) with or without an active component of liposome size from 20 to 500 nm, having a double (multilayer) layer in the structure around the liquid core, which allows them to encapsulate both lipophilic active substances and hydrophilic active substances.

The advantages of the present invention include the fact that:

The composition may be a carrier of medicinal substances, which, upon direct contact with the skin, cause an irritating effect.

The shell and structure of liposomes (nanosomes) allows you to encapsulate biologically active substances and medicinal substances of various chemical structures and physical properties, both lipophilic and hydrophilic, taking into account pharmaceutical compatibility at the same time.

High bioavailability of the composition and preparations based on it, taking into account the approximate structure of nanosome membranes and cell membranes.

The membrane of liposomes (nanosomes) increases the resistance of the active substances encapsulated inside the liposome to adverse environmental factors and ensures the preservation of their activity, increasing the shelf life of the drug based on the composition.

Liposomes (nanosomes) themselves, containing or not containing the active component, do not cause irritating effects on the skin, which allows them to be used to obtain transdermal preparations.

Liposomes themselves, without the encapsulated active ingredient, additionally have a moisturizing and soothing effect on the skin.

The composition provides for selectivity of the effect of the active compound on body tissues.

The composition allows you to adjust the rate of penetration of the active substance through the skin by affecting the flexibility of the nanosome membrane. More flexible liposomes are characterized by faster penetrating speed. The flexibility of liposomes is controlled by placing liposomes in an appropriate medium: in a medium with a high water content (spray, lotion, tonic, gel, etc.), nanosomes penetrate faster; liposomes in an environment with an increased content of lipid fraction (ointment or gel base, including the additional introduction of oils) slows down the rate of penetration of liposomes through the cell membranes of the body and this property can be used for drugs designed for a long time of introduction into the body, adjusting the time effects on the tissue.

The composition allows you to dose the active components and limit their upper limit, which is important for potent substances. For active substances with hydrophilic properties, the maximum concentration of substances included in liposomes (nanosomes) is up to 30%, for active substances with lipophilic properties, up to 25%.

The composition is designed for direct human use and / or for the preparation of a medicine, cosmetic, specialized food product or biologically active food supplement. The most optimal use of the composition is for the preparation of external dosage forms (gels, creams, ointments, emulsions, sprays, etc.). To this end, the composition can act as a pre-prepared semi-finished product, which simplifies the scheme for obtaining the final product. So, in the composition of ointments, the composition is introduced at the final stage of production with continuous stirring at a temperature of not more than 30 ° C.

For emulsion type preparations, when an liposome (nanosome) is introduced into the formula, an emulsifier may be used. The most optimal use of non-ionic emulsifiers type M / B, anionic emulsifiers type M / B, non-ionic emulsifiers type B / M, anionic emulsifiers type B / M.

An essential distinguishing feature of the claimed composition is the presence in the composition of liposomes (nanosomes) with a double layer of membranes (multilayer), as well as the possibility of incorporating biologically active substances and medicinal substances into liposomes (nanosomes) regardless of their properties and solubility, while the average diameter of liposomes ( nanosome) is from 20 to 500 nm.

The invention is illustrated by the following examples of industrial implementation in a production environment.

Examples of specific performance.

Example 1

Ready-to-use form - composition of liposomes (nanosom) without filler (based on 100 wt.%):

Ethanol 1,0 Phospholipids (Lecithin) 1,0 (with a phosphatidylcholine content of at least 60%) Water 98.0

A solution of lecithin in ethanol is prepared, the solution and deionized water are placed in a reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it. The resulting preparation is analyzed for compliance with the main quality criteria (particle size, description, odor, pH value and microbiological purity). The mass of liposomes (nanosomes) is stored in appropriate tightly closed containers, protecting from exposure to light at a temperature of + 4 ° C, avoiding microbiological coamination. The resulting composition is used as a semi-finished product for the production of medicines, cosmetics and food products.

Example 2

Ready-to-use form - composition of liposomes (nanosom) without filler (based on 100 wt.%):

Ethanol 30,0 Phospholipids (Lecithin) 30,0 (with a phosphatidylcholine content of at least 60%) Water 40,0

A solution of lecithin in ethanol is prepared, the solution and deionized water are placed in a reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it. The resulting preparation is analyzed for compliance with the main quality criteria (particle size, description, smell, pH of the medium and microbiological purity). The mass of liposomes (nanosomes) is stored in appropriate tightly closed containers, protecting from exposure to light at a temperature of + 4 ° C, avoiding microbiological coamination. The resulting composition is used as a semi-finished product for the production of medicines, cosmetics and food products.

Example 3

Ready-to-use emulsion based on a composition of liposomes with coenzyme Q10 (based on 100 wt.%):

Ethanol 10.0 Phospholipids (Lecithin) 5,0 (with a phosphatidylcholine content of at least 60%) Fatty Acid Triglycerides 20,0 Coenzyme Q10 (ubiquinone) 5,0 Water 60.0

A solution of lecithin in ethanol is prepared, coenzyme Q10 is separately dissolved in the lipid fraction (triglycerides of fatty acids), the medium is prepared separately - deionized water, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel forms in it, emulsify for 30-40 minutes. The resulting preparation is analyzed for compliance with the main quality criteria (particle size, description, odor, pH, microbiological purity, quantitative content of the active component). The resulting product does not have a general toxic, irritating and allergenic effect, has a uniform consistency, is evenly applied to the skin, and has an antioxidant effect. The technical result is to increase stability and increase the shelf life of the emulsion for external use, as well as improving the bioavailability and selectivity of the active compounds.

Example 4

A ready-to-use form is a gel containing a composition of liposomes (nanosomes) with the active substance in a concentration of 0.001% (based on 100 wt.%):

Resveratrol Liposomes 10.0 including: ethanol 1,5 phospholipids (lecithin) 2.5 (with a phosphatidylcholine content of at least 60%) resveratrol 0.001 water 6.0 Nipagin 0.1 Nipazole 0.1 Gel crosslinked polyethylene oxide-1500 up to 100.0

A solution of lecithin in ethanol is prepared, resveratrol in water is separately dissolved, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes. The resulting mass is introduced into the gel base at a temperature not exceeding 30 ° C, preservatives (nipagin and nipazole) are added and mixed for 10-20 minutes.

The resulting product does not have a general toxic, irritating effect, has a uniform consistency, is evenly applied to the skin, and has antioxidant and moisturizing effects.

Example 5

A ready-to-use form is a gel containing a composition of liposomes (nanosomes) with the active substance in a concentration of 30% (based on 100 wt.%):

Resveratrol Liposomes 90.0 including: ethanol 9.0 phospholipids (lecithin) 9.0 (with a phosphatidylcholine content of at least 60%) resveratrol 30,0 water 42.0 Nipagin 0.1 Nipazole 0.1 Gel crosslinked polyethylene oxide-1500 up to 100.0

A solution of lecithin in ethanol is prepared, resveratrol in water is separately dissolved, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes. The resulting mass is introduced into the gel base at a temperature not exceeding 30 ° C, preservatives (nipagin and nipazole) are added and mixed for 10-20 minutes.

The resulting product does not have a general toxic, irritating effect, has a uniform consistency, is evenly applied to the skin, and has an antioxidant effect.

Example 6

Ready-to-use ointment based on a composition of liposomes with diclofenac sodium (based on 100 wt.%):

Liposomes containing diclofenac sodium 10.0 In terms of diclofenac sodium 1,0 including: ethanol 1,0 phospholipids (lecithin) 2.0 (with a phosphatidylcholine content of at least 60%) diclofenac sodium 1,0 water 7.0 Polyethylene Oxide-400 20,0 Polyethylene Oxide-4000 20,0 1,2-propylene glycol 49.8 Nipagin 0.1 Nipazole 0.1

A solution of lecithin in ethanol is prepared, sodium diclofenac is separately dissolved in water, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes. The resulting mass is introduced into the ointment base at a temperature not exceeding 30 ° C, preservatives (nipagin and nipazole) are added and mixed for 10-20 minutes.

The resulting product does not have a general toxic, irritating effect, has a uniform consistency, is evenly applied to the skin, and has an anti-inflammatory effect. The technical result is an ointment for external use with improved bioavailability and selectivity of diclofenac, increasing the safety of the drug.

In practice, you can make changes and modify the described preferred embodiments of the present invention, without departing from the subject and objectives of the present invention, within the scope of the claimed claims.

Studies have been conducted to study the specific activity of the drug according to example 6.

The anti-inflammatory activity of an ointment based on sodium diclofenac liposomes was studied on a model of rat kaolin edema. The experiments were carried out on white outbred rats kept in a vivarium under stationary conditions on a standard diet. Each group of animals in the experiment consisted of 6 rats weighing 170 ± 20.0. Inflammation was caused by the administration of 0.1 ml of a 10% suspension of kaolin under the hind paw aponeurosis. The effect of ointment with diclofenac sodium liposomes on the second stage of inflammation, exudation, was determined. Edema was evaluated oncometrically, its value was determined by the change in the volume of the legs, which was measured before the introduction of flagogen, as well as 5, 24 and 48 hours after the introduction of a suspension of kaolin. The effectiveness of the studied composition was compared with the effectiveness of the classic ointment of diclofenac sodium 1% (Mashkovsky M.D. Medicines. M., 2005. T. 1, 2; composition: 1 g of the ointment contains 10 mg of diclofenac sodium, excipients - polyethylene oxide-400, polyethylene oxide-4000, 1,2-propylene glycol, nipagin, nipazole).

The first group of animals did not receive treatment, the second group was applied diclofenac ointment 1%, the third - gel base, the fourth - ointment composition with diclofenac sodium liposomes 1%. The results obtained are presented in table 1. As follows from the results obtained, the ointment with diclofenac sodium liposomes by pharmacological activity was more effective than the prototype (classical diclofenac ointment 1%).

Based on the foregoing, it can be concluded that the proposed composition has several advantages in comparison with the prototype and will find wide application in obtaining effective funds.

Example 7

Liposomes containing fucoidan, escin and caffeine: Ethanol 10.0 Phospholipids (Lecithin) 5,0 (with a phosphatidylcholine content of at least 60%) Fatty Acid Triglycerides 20,0 Fucoidan 5,0 Escin 5,0 Caffeine 10.0 Water up to 100.0

A solution of lecithin in ethanol is prepared, fucoidan, escin and caffeine are separately dissolved in water, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes. The resulting mass is collected in a container and further used to obtain finished dosage forms by adding the calculated amounts to the compositions. This phospholipid composition with accurate amounts of active substances allows you to dose the pharmacologically active component in effective amounts (example 8).

Example 8

Liposomes containing fucoidan, escin and caffeine 5,0 including: ethanol 0.5 phospholipids (lecithin) 0.25 (with a phosphatidylcholine content of at least 60%) fatty acid triglycerides 1,0 fucoidan 0.25 escin 0.25 caffeine 0.5 water 2.25 Nipagin 0.1 Nipazole 0.1 Gel crosslinked polyethylene oxide-1500 up to 100.0

So, the resulting mass in Example 7 in an amount of 5% is introduced into the gel base at a temperature of no higher than 30 ° C, preservatives (nipagin and nipazole) are added and mixed for 10-20 minutes.

This composition was subjected to in vivo testing. The study involved 5 volunteers aged 30 to 50. The composition was applied around the eye area daily 1 time per day. Course duration - 14 days. Studied target parameter: skin color (chromameter). At the end of the study, the skin condition was checked. On day 14, all 5 participants showed a significantly lighter area around the eyes compared to the start of the study. Figure 1 shows a photograph of the eye area of one of the participants in the study.

The average data after statistical processing are presented graphically in Fig. 2.

Example 9

Liposomes containing linoleic acid, vitamin E, D-panthenol and caffeine 15.0 including: ethanol one phospholipids (lecithin) 0.5 (with a phosphatidylcholine content of at least 60%) fatty acid triglycerides 1,0 linoleic acid 0.5 vitamin e 0.5 D-panthenol 0.5 water up to 15.0 Excipients for hair emulsions up to 100.0

A solution of lecithin in ethanol is prepared, linoleic acid, vitamin E and D-panthenol are separately dissolved in water, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes. The resulting mass is introduced into the emulsion base at a temperature not exceeding 30 ° C and stirred for 10-20 minutes.

This example was used to assess the ability to control the flexibility, mobility, penetration and stability of the composition, which was confirmed by studies in the treatment of hair loss.

Due to the structure of liposomes, the highest degree of penetration of active substances and their delivery to the follicles was achieved, which provided a stop to hair loss and restoration of problem areas. The study involved 11 volunteers (6 men and 5 women). For 24 weeks, 4 people were daily applied in the form of a hair mask with a product containing vitamin E, linoleic acid and D-panthenol in concentrations of 1%, 2.5% and 3%, respectively.

The remaining participants used a topical emulsion for hair, which was applied to the scalp for 20 minutes, after which the emulsion was washed off with a sufficient amount of water.

At the beginning and end of the study, the average amount of hair in the problem areas (number of hair / cm ² ) and the condition of the hair were calculated.

At the end of the study, it was shown that in patients using a well-known product that does not contain active substances in liposomal form, there were no visible changes to improve the condition of the hair. Participants using liposome composition showed a significant improvement and restoration of problem areas (Fig. 3.)

Table 1 The effect of ointment with diclofenac sodium liposomes on kaolin edema A drug The average increase in edema,% after 5 hours after 24 hours after 48 hours The control 107.8 ± 8.4 85.9 ± 6.9 64.6 ± 8.6 Diclofenac Sodium Ointment 1% 83.7 ± 11.5 59.2 ± 9.2 * 46.7 ± 8.5 * The basis of model ointments 98.1 ± 6.3 80.0 ± 6.2 61.9 ± 5.9 Diclofenac Sodium Liposome Ointment 74.4 ± 9.2 * 43.6 ± 17.3 * 24.1 ± 10.2 * Note: * - the value is statistically significantly different from the control at P <0.05.

The resulting composition according to example No. 6, containing diclofenac, was used to study the stability of the structure.

Diclofenac, enclosed in liposomes, was placed in the form of a gel. To increase the rate of ESR determination, vitamin E was additionally placed on liposomes. The penetrating power of the composition (blue curve) was compared with a commercially available ointment (blue curve).

The natural antioxidant potential of the skin was taken as 100%. A rising curve shows increased penetration. An interesting fact discovered is that a commercially available ointment not only does not show penetration (to be described by a straight line at 100%), but also acts as an oxidizing agent (damages the skin).

The composition according to example No. 6 demonstrates excellent penetration after 30 minutes after application.

The carrier system (liposomes) was supplemented with a 14C-labeled diclofenac and was used for skin biopsy. After 8 hours, the upper layer was removed from the skin 16 times, and the count of radioactive parameters was measured in the layers of the skin, as well as in adipose tissue and in the liquid. Using commercially available ointments, studies were compared.

The carrier system shows very good penetration, while commercial ointments are mainly found in the top 4 strips.

Based on the proposed phospholipid composition, most of the diclofenac used was directed to the skin.

Moreover, diclofenac also penetrated into adipose tissue, which shows the trans-dermal penetrating ability of the carrier system. This in turn can facilitate the accumulation of diclofenac in the joints.

In animal tests, liposomally encapsulated diclofenac (a test sample codenamed Cutisome No Pain) was compared with commercially available diclofenac ointments and tablets. It has been shown that the proposed carrier system guarantees an excellent supply of muscle and kneecap with diclofenac, which is superior to larger doses of diclofenac for oral use.

Moreover, it was shown that the penetrating ability of the carrier system also exceeds commercially available ointments and tablets, despite the fact that the concentration of the drug in the carrier system is 10 times lower (Fig. 4).

Example 10

Liposomes containing lutein extract:

Ethanol 10.0 Phospholipids (Lecithin) 5,0 (with a phosphatidylcholine content of at least 60%) Lutein Extract 25.0 Water up to 100.0

A solution of lecithin and lutein in ethanol is prepared, water is added separately, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes.

We obtained 4 prototypes of nanosystems with lutein extract according to this example using lutein with different variants of phosphatidylcholine content. The biopharmaceutical properties of nanosystems were evaluated by the speed and completeness of dissolution in an in vitro test.

At the first stage, the rate of release of water into the dissolution medium was evaluated: ethyl alcohol (7: 3 by volume), recommended in the pharmacopeia article for substances poorly soluble in aqueous media.

The release rate was estimated using the “Dissolution” test on a “rotating basket” type instrument (Type DT 600, Erweka, Germany) at a temperature of (37 ± 1) ° С. The rotation speed is 100 rpm. A sample of the product, equivalent to 20 mg of lutein extract, was placed in a basket and after specified periods of time (5, 15, 30, 45, 60, 90 minutes and 2, 3, 4 hours) samples were taken.

HPLC was used for the quantitative analysis of lutein in the release medium. The analysis was performed on a Shimadzu high-pressure chromatograph (Japan) with a Luna C18 column (4.6 × 150 mm, 5 μm) and a pre-column (2.0 mm) filled with the same sorbent (Phenomenex, USA) in isocratic elution with a methanol: acetonitrile 5 mixture : 95 (by volume), eluent flow rate of 1 ml / min, dosed sample volume of 20 μl, detection wavelength of 450 nm.

Chromatogram registration and processing was performed using LCSolution software (Shimadzu, Japan).

Dosing of all samples was carried out at least twice. The calculation was carried out according to a calibration curve constructed using methanol solutions of lutein extract. The amount of lutein transferred to the solution was calculated, in% of the maximum possible.

Statistical processing of chromatographic data was performed using Origin 4.1 software.

The experimental results are shown in Fig. 5.

During the experiment, it was found that the introduction of lutein extract in the composition of L3 practically does not affect the rate of release. From the disperse system L4, the amount of released lutein reached 38% by the 30th minute of the experiment, however, no further release was observed. The dispersed system L2 allows to release about 50% of lutein by 2 hours of the experiment. The best lutein release is observed from the L1 system: by the 60th minute of the experiment, about 65% is released.

The dissolution rate constants calculated by the least squares method were:

L1 composition - 0.5012 min ^-1

L2 composition - 0.3435 min ^-1

L3 composition - 0.0449 min ^-1

L4 composition - 0.2234 min ^-1

lutein extract - 0.02798 min ^-1

Thus, the greatest completeness and rate of lutein release is observed from formulations L1 and L2.

For these samples, the rate of release into a two-phase dissolution medium was studied, which models the conditions of the gastrointestinal tract for lipophilic, water-insoluble substances and correlates well with in vivo pharmacokinetics.

The in vitro release rate was estimated using the Dissolution test on a rotating mixer type instrument (Type DT 600, Erweka, Germany) at a temperature of (37 ± 1) ° С. The rotation speed is 100 rpm, the dissolution medium is a two-phase water: octanol system 3: 1, the total volume of the dissolution medium is 400 ml.

For the quantitative analysis of lutein in the release medium, direct UV spectrophotometry was used. The optical density of the obtained solutions was measured on a Shimadzu spectrophotometer at a maximum wavelength (450 nm) in a cuvette with a layer thickness of 1 cm.

The amount of lutein transferred to the solution (in% of the maximum possible) was calculated using the extinction coefficient E1% 1 cm = 2560.

The results are shown in Fig. 6.

The release proceeds most quickly and completely from the dispersed system L1: by the 30th minute of the experiment, the maximum release is reached, which is about 43%, which is 14 times more compared to the pure lutein extract (about 3%). The maximum concentration upon release from L2 is reached by 2 hours and does not exceed 26%. The dissolution rate constants calculated by the least squares method were:

L1 composition - 0.4450 min ^-1

L2 composition - 0.3499 min ^-1

lutein extract - 0.0424 min ^-1

The data obtained allow us to conclude that the introduction of lutein extract into nanosystems can significantly increase its bioavailability in vitro. Of the obtained nanosystems, the dispersed system L1 is characterized by the highest bioavailability.

The average particle size determined by photon correlation spectroscopy on a Malvern Zetasizer 3000HSA instrument (Malvern instruments, Worcestershire, UK) was 152 ± 2 nm.

Example 11

Liposomes containing blueberry extract:

Ethanol 10.0 Phospholipids (Lecithin) 5,0 (with a phosphatidylcholine content of at least 60%) Blueberry extract 20,0 Water up to 100.0

A solution of lecithin in ethanol is prepared, water and blueberry extract are added separately, everything is placed in a single reactor with a stirrer, the mass is untwisted until a stable rotation funnel is formed in it, emulsified for 30-40 minutes.

We obtained 2 prototypes of nanosystems with blueberry extract using lecithin with different phosphatidylcholine content. The content of blueberry extract in nanosystems was 20 wt.%.

For the obtained samples, the release rate in the test was studied in vitro on a rotating basket instrument (Type DT 600, Erweka, Germany) at a temperature of (37 ± 1) ° С. The rotation speed is 100 rpm, the dissolution medium is 0.1 n. hydrochloric acid, the volume of the dissolution medium is 250 ml. A portion of the test product (about 20 mg in terms of blueberry extract) was placed in a basket. After the set time intervals (3, 5, 10, 15, 20, 30, 40, 50, 60 min), 1 ml samples were taken, the volume was replenished with the same solvent.

For quantitative analysis of anthocyanins, the direct spectrophotometry method was used. The optical density of the obtained solutions was measured on a Shimadzu spectrophotometer at a wavelength of maximum (510 nm) in a cuvette with a layer thickness of 1 cm. The number of anthocyanins transferred to the solution was calculated, in% of the maximum possible.

The experimental results are shown in Fig. 7.

The data obtained allow us to conclude that the release from the extract itself and composition B1 have practically no differences: by the 5th minute of the experiment, the maximum concentration in the release medium is reached, the amount of released anthocyanins is about 90%. System B2 is characterized by a slower release: maximum release is achieved by the 30th minute of the experiment and is about 95%.

The dissolution rate constants calculated by the least squares method were:

B1 composition - 7.6900 min ^-1

B2 squad - 2.9746 min ^-1

blueberry extract - 8.3207 min ^-1

The data obtained allow us to conclude that the introduction of blueberry extract into the dispersed B1 system does not affect its bioavailability in vitro, the B2 system provides a more prolonged release while maintaining a high level of bioavailability. The considered models of in vitro experiments do not allow us to show the advantages of nanosystems with blueberry extract due to the high bioavailability of the extract itself. More adequate data on increasing the bioavailability of products based on blueberry extract can be obtained only after studying the pharmacokinetics in vivo, for example, in rabbits.

The average particle size determined by photon correlation spectroscopy was 268 ± 15 nm.

During crystallization of a solid from a solution or melt, the least stable phase is formed first, which is closest to the solution in terms of free energy (V. Ostwald rule).

That is why metastable modifications have less internal adhesion of molecules, which is expressed in their increased solubility. The solubility of a drug substance in water seems to be its most important physical property.

Dissolution plays the role of a limiting stage and absorption of the drug from the gastrointestinal tract. That is why it is precisely metastable forms that are of particular interest when creating a pharmaceutical substance.

Comparing the lutein dissolution rate constants, we can conclude that nanosystems are characterized by a faster release of water-insoluble lutein compared to a substance. So the rate of release of lutein from the nanosystem increases by 10.5 times. Thus, the introduction of lutein extract in the composition of the nanosystem indicates a significant improvement in its release in the aquatic environment. This effect can be explained by a decrease in particle size and stabilization of particles in the composition of the nanosystem.

Thus, the introduction of plant extracts into the composition of the metastable form based on nanosystems can significantly increase the rate of their dissolution and ensures uniform release of the active substance under the conditions of oral administration.

Claims (13)

1. Phospholipid emulsion comprising liposomes (nanosomes) with an average diameter of liposomes (nanosomes) from 20 to 500 nm, containing ethanol and water, characterized in that it additionally contains lecithin with a phosphatidylcholine content of at least 60% and diclofenac sodium in the following ratio of liposome components and active substance, wt.% (per 100 wt.%):
ethanol 1.0-30.0% phospholipids (lecithin) 1.0-30.0% diclofenac sodium 0.001-30.0% water up to 100.0% 2. The phospholipid emulsion according to claim 1, containing at least one acceptable auxiliary component as a diluent. 3. The phospholipid emulsion according to claim 1, characterized in that the emulsion allows you to dose the pharmacologically active component in effective quantities and limit their upper limit. 4. The phospholipid emulsion according to claim 1, characterized in that it contains at least one of the excipients selected from the groups of preservatives, aromatic composition. 5. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one vitamin and / or vitamin-like substances. 6. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one extract. 7. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one micro or macro element. 8. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one compound from the group of carotenoids. 9. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one compound from the group of organic acids. 10. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one compound from the group of enzymes. 11. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one compound from the group of flavonoids. 12. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises at least one compound from the group of chondroprotectors. 13. The phospholipid emulsion according to any one of claims 1 to 4, which further comprises resveratrol.

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