RU2381792C2 - Mesotherapy preparation - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to cosmetology and dermatology and represents a mesotherapy preparation characterised by that it contains an aqueous emulsion of perfluororganic compounds concentrated therein 5-95 vol %.

EFFECT: invention provides development of a new mesotherapeutic preparation with high efficiency when used in small dosage, and prolonged action.

2 cl, 4 tbl, 5 dwg

Description

The invention relates to the field of cosmetology and dermatology and relates to the injection method of intradermal and subcutaneous administration of various allopathic and homeopathic preparations - mesotherapy.

The term mesotherapy was proposed in 1958 by the French physician Michel Pistor. Mesotherapy is used in classical medicine in the complex treatment of various diseases: musculoskeletal system, ENT pathology, gynecology, urology, neurology, rheumatology, orthopedics, dentistry; in the complex treatment of dermatological diseases, as well as in aesthetic medicine for solving various face and body problems: age-related skin changes (wrinkles, decreased skin tone and elasticity), hyperpigmentation, pasty and swollen skin, atrophic, hypertrophic and keloid scars, erythrosis, rosacea, varicose veins, cellulitis, excess local fatty deposits on the face ("second chin") and the body, alopecia. Mesotherapy is one of the most effective skin rejuvenation methods.

Mesotherapy got its name from the injection site - the mesofliss layer of the skin, where the blood vessels are concentrated, as well as tissues that determine the external condition of the skin. Mesotherapy is not an alternative treatment to traditional medicine, as it might seem at first glance. This method involves the introduction of well-known and well-established in the medical practice of medicines and nutrients. At the same time, either the entire painful area or a separate anatomical site is chipped, or the drug is administered at certain points.

Today in mesotherapy there are several areas that do not exclude each other. With the allopathic method, traditional medicines are introduced - vitamins, antibiotics, local anesthetics, etc., made from plant material. With the homeopathic method of mesotherapy, respectively, homeopathic preparations are introduced - these are introductory solutions that do not contain oily substances. To work with women over forty use biologically active substances of animal origin. They contain extracts of animal fetal tissue. They are used to create a rejuvenating effect. The drug may also contain proteins, acids, vitamin and mineral complex.

Means for mesotherapy can be either ready-made, that is, made in the factory, or assembled from individual components immediately before administration. The last way of obtaining involves the doctor’s ability not only to administer the drug, but also knowledge of the formulations and the interaction of various active principles. Most often, mesotherapeutic cocktails include B vitamins, vasodilators, blood flow improvers in small vessels, hyaluronic acid and elastin, as well as many others.

There are different types of mesotherapy and tools for its implementation.

The mesotherapy procedure solves a large number of problems in various areas of the face and body, in particular with face mesotherapy, the signs of skin aging are prevented and eliminated, the complexion and skin tone are improved, wrinkles can be removed for a certain time, pigmentation is eliminated, and skin oilyness is reduced. During body mesotherapy, skin is strengthened, cellulite can be treated and local fat deposits removed, blood circulation improves, blood vessels strengthen.

A large number of funds for mesotherapy are known (O.S. Ozerskaya. Technological foundations of mesotherapy (intradermal administration of drugs), S / P, 2007).

Known, for example, a number of drugs company "Revitacare sarl" (France). Among them are:

- “Cellulare” containing hyaluronic acid, caffeine-benzoate sodium, manganese, zinc, cobalt gluconates, which has a lipolytic, antioxidant and regenerative effect;

- "Haircare" containing vitamins B3, B5, B6, B8, B9, B12, arginine, cysteine, glutamine, glycine, ornithine, zinc gluconate and improves microcirculation;

- "Stretchcare" containing DMAE, hyaluronic acid, vitamin B5, copper and zinc gluconates. The complex has a regenerating and anti-inflammatory effect.

Known drug "DMAE-MONO" (Alianza, Brazil), containing 6% dimethylaminoethanol, regenerating and stimulating connective tissue.

The same company produces the drug "Mezovasco" containing sodium heparin and buflomedil. The drug has a vasodilating and disaggregating effect.

The company "Alianza" (Brazil) also produces a number of drugs based on natural raw materials. See table 2.

A well-known complex created in France by the Filorga Laboratory, which is based on the use of biologically active agents (embryoblasts) to slow down the aging process of the skin. The Embryoblast complex rejuvenates the skin, restores and renews its structure, stimulates the formation of collagen, does not cause the effect of addiction and withdrawal syndrome. Contains vitamins, minerals, amino acids, nucleic acids (DNA, RNA), biostimulins, cell growth factor. Restores the natural regeneration of the skin. Indicated for the prevention and correction of age-related skin changes (wrinkles, decreased tone, turgor) Embrioblast is a leading drug in anti-age therapy.

The technical task of the present invention is the creation of a new mesotherapeutic agent with high effectiveness when using lower doses of the drug and increasing the prolongation of its action.

This technical problem is solved by creating a new tool for mesotherapy, containing an aqueous emulsion of perfluororganic compounds (PPS).

The optimal concentration of perfluoroorganic compounds in the emulsion is 5-95 vol.%. In this case, the emulsion of perfluoroorganic compounds can be direct, reverse or multiple, and as the second phase contain not only water, but also / or glycerin and / or aqueous solutions of carbohydrates.

The mesotherapy agent may additionally contain biologically active substances, such as: water-soluble compounds, for example, vitamins, and / or antioxidants, and / or vasoactive substances, and / or lipolytic substances, and / or biostimulating substances, and / or non-steroidal anabolic substances, and / or corticosteroid substances and / or enzymes and / or antibacterial and antiviral substances and / or immunomodulators and / or gas exchange modulators.

As a gas exchange modulator, alkali metal hydrogen carbonates and / or organic phosphates can be used.

As organic phosphates, it is preferable to use 2, 3-diphosphoglycerate and / or hexainositol phosphate.

The tool may additionally contain substrates of biosynthesis of nitric oxide, and, in addition, biologically active substances, which are fat-soluble compounds in the form of dispersed systems, for example, emulsions and / or micellar solutions.

The use of PFC emulsions in medicine and cosmetics has long been well known (JGRiess, Oxygen Carriers, Chem. Rev., 101 (9), 2797-2920, 2001; patent RU 2033163 C1 (JSC NIZAR), 1995.04.20, A61K 7/00, 33/16).

As a result of numerous studies conducted in various countries, it was shown that the use of PPS emulsions in medicine for internal and external use provides:

- dissolution and transfer of vital gases (O ₂ , CO ₂ , NO)

- accelerated delivery of oxygen to tissues

- improvement of metabolism and gas exchange at the tissue level

- restoration of central hemodynamics

- improvement of peripheral blood flow and microcirculation

- stabilization of cell membranes

- cardioprotective effect

- activation of detoxification function of the liver

- sorption and diuretic effect

- immunomodulatory effect

- wound healing and anti-burn effect.

The pioneers of incorporating PPS emulsions into various cosmetics were NIZAR (Russia) and Lancaster (USA), for which corresponding patents were obtained in the nineties and beyond. It was shown that the drug "Nizaftem" (brand), containing an emulsion of perfluororganic compounds (company NIZAR, RF), performs the following:

- transfers vital oxygen to skin tissue

- stimulates the processes of tissue respiration

- activates tissue regeneration processes

- stimulates the metabolism in skin tissues

- increases the content of collagen and elastin in skin tissues

- reduces keratin content

- improves the penetration of water and biologically active substances into the skin

- stops irritations

- enhances the action of sunscreens and increases SPF.

The authors suggested that all these properties of PPS emulsions are preserved during mesotherapy. However, the authors did not find sources of information containing information about mesotherapy preparations containing PPS emulsions.

Further experiments confirmed the possibility of using and the high efficiency of the preparations proposed by the authors.

In the course of experiments to study the effectiveness of mesotherapy drugs, the means of PPS emulsions of various compositions were investigated. Emulsions were prepared using high pressure homogenization techniques. All studied preparations were isoosmotic and had a pH of 6.0-7.4 with a particle size of 70-300 nm (according to laser photon correlation spectroscopy). The composition of the drugs and their properties are described in Table 3.

To confirm the high efficiency of the use of products containing perfluorocarbon emulsions, studies have been conducted in mesotherapy practice that demonstrate the high biological activity of such emulsions upon intradermal administration.

In particular, by the example of the NIZAFTEM ^® -N emulsion, the effect of intradermal administration of PPS preparations on skin oxygenation was studied. The experiment was conducted on 6 Balb / C mice using the ESR method of oximetry. Lithium phthalocyanine, a suspension of which was injected subcutaneously, was used as an oxygen sensitive paramagnetic probe. 4 days before the start of the experiment, the hairline was removed at the injection site. The NIZAFTEM ^® -N emulsion was administered once intradermally in a total dose of 10 μl. 2.5 μl to the centers of 4 squares 0.5 cm × 0.5 cm, covering a skin area of 1 cm × 1 cm. The measurements were carried out in a thermostatic box at 36.5 ° C at an operating frequency of the ESR spectrometer of 1.1 GHz.

Figure 1 presents comparative data on the study of changes in pO ₂ in the skin with the introduction of various preparations of standard physical. a solution and a 4% dispersion of liposomes from soya lecithin containing 0.05% nicotinic acid with the addition of 2.6% sorbitol and NIZAFTEM ^® -L.

As can be seen from the graph (figure 1), the intradermal administration of the PPS emulsion can significantly increase the partial pressure of oxygen in the skin and ensure its long-term effect.

To demonstrate the higher intensity PFCs emulsions exposure at mesotherapeutic injections compared to epicutaneous application was studied pharmacokinetics of perfluorodecalin (PFD) in the skin of Wistar rats at the intradermal injection of emulsions "PERFTORAN ^®» (10% PPS by volume) and epicutaneous application emulsion "NIZAFTEM ^® -N ”(40% SFC by volume). The proportion of PFD of the total number of PPS is in the first case 70%, and in the second 85%. The emulsion "-H NIZAFTEM ^®" was applied to the shaved back of the animal surface of a single dose of 0.1 g per cm ² on the portion 2 cm × 2 cm and rubbed fingers until absorbed. The PERFTORAN ^® emulsion was injected in an amount of 10 μl intradermally once with a microsyringe into the center of the same skin area of other animals. Rats after the application of NIZAFTEM ^® were sacrificed after 0.25, 0.5, 2, 4, 6, and 8 hours, and rats after injection of "PERFTORAN ^® " - after 0.25 hours and on days 1, 7, 14, 21, 30. A 500 mg piece of skin (approximately 1.5 cm × 1.5 cm × 0.2 cm) was thoroughly wiped with filter paper and subjected to destruction in a solution of proteolytic enzymes. PPS contained in tissues was extracted from the obtained lysate using 0.5 ml of an emulsion containing 20% fluorocarbon C ₆ F ₁₃ CH = HCF ₁₃ C ₆ , characterized by high purity and long retention time on a chromatographic column. After 2 hours of extraction, the emulsion was destroyed by the addition of an equal volume of ethyl alcohol. A drop of separated fluorocarbons was precipitated by centrifugation and washed several times with water, after which it was used for chromatographic analysis. Quantitative determination of PFD in the samples was carried out on a Chrom-5 chromatograph with a glass column (2 m × 2.5 m) filled with 25% OV-1 on a Gas Chrom Q, 80-100 mesh. The detector is flame ionization, the carrier gas is nitrogen, the pressure at the column outlet is 0.6 atm, the temperature of the dispenser, thermostat and detector are 160, 60, 120 ° С, respectively.

According to the results shown in the graph (figure 2), with a significantly smaller amount of PPS emulsion in the amount of emulsion used for processing, with intradermal administration of the PERFTORAN ^® emulsion, a significantly higher content of PFD in the skin is observed. Even a month later, it exceeds the maximum PFD content observed 0.5 hours after the application of the NIZAFTEM ^® -N emulsion (Fig. 2A). Thus, with single mesotherapy injections of PPS emulsions, there is a prolonged effect of these biologically active substances on the skin, which contributes to the long-term positive effect of the corresponding cosmetic procedures. This conclusion is confirmed by the results of studies of changes in the biosynthesis of major skin proteins, given below.

To demonstrate the rejuvenating effect of PPS emulsions and the high efficiency of mesotherapeutic procedures, they were used to study changes in the biosynthesis of keratin, collagen and elastin in the skin of BALB / c mice of different age groups and for different applications of PPS emulsions. As young animals, mice were used at the age of 2 months, middle-aged animals at the age of 10 months, old animals at the age of 20 months. For studies of the effect of PFC emulsions, middle-aged animals were used. Each experimental group included 6-10 mice. Before the experiment, hair was removed from the skin exposed to experimental exposure. The NIZAFTEM ^® -N emulsion was applied, as described above, daily for 30 days to a 1 cm × 1 cm area. The PERFTORAN ^® emulsion was administered once intradermally in a total dose of 10 μl, 2.5 μl, to the centers of four squares of 0.5 cm × 0.5 cm covering the skin area 1 cm × 1 cm equivalent to the application area “NIZAFTEM ^® ”. To control the effect of manipulations, we used a group of animals whose skin was treated with 0.8% CARBOPOL ^® gel as described for the NIZAFTEM ^® -N emulsion. In control animals of different age groups, an equivalent area of skin was shaved, which was not exposed to any further effect. Control animals and animals exposed to the CARBOPOL ^® gel and the NIZAFTEM ^® -N emulsion were killed on the 30th day from the beginning of the experiment, and animals after the injection of the PERFTORAN ^® emulsion on the 30th and 60th days. Treated skin areas were excised from slaughtered animals and the epidermis and dermis were mechanically separated. The total keratin content was determined in the obtained epidermal samples, and the content of water-soluble collagen and elastin in the dermis samples.

In accordance with the results shown in the graphs (figure 3-5) when exposed to PFC emulsions, there is a shift in the biosynthesis of marker proteins to indicators of young animals. So, in the cells of the epidermis when exposed to these emulsions, keratin synthesis increasing with age is significantly reduced (Fig. 3). Moreover, a single mesotherapy injection compared with multiple cutaneous applications during the month gives a stronger and longer lasting effect, which lasts at least two months. The synthesis of structural proteins of the dermis — collagen (FIG. 4) and elastin (FIG. 5), decreasing with age, when exposed to PPS emulsions, on the contrary, increases markedly. At the same time, as in the case of keratin, the anti-aging shift is more pronounced and stable during mesotherapeutic exposure.

A comparative analysis of the effectiveness of perfluorocarbon emulsions and known mesotherapy agents was made in experiments on rats using PERFTORAN ^® emulsion and the widely used Embryoblaste mesotherapy drug (Laboratoires Filorga, France). The effect of mesotherapeutic procedures on the elasticity of the skin of the heel region of the hind limbs of rats was studied against the background of the intensification of aging by ultraviolet radiation. Skin elasticity was measured with a SEM 575 cutometer (Courage + Khazaka electronic GmbH, Germany) using a 2 mm diameter sensor. The measurement suction / recovery cycle included suction for 3 s at 500 mbar, followed by removal of negative pressure for 3 s. Using the obtained dependences of the length of the stretched fragment on time, the elasticity parameters were measured: the value of fast pulling (U _e ), measured by 0.1 s from the beginning of the cycle; the value of slow tightening (U _v ), equal to the increase over time from 0.1 to 3 s; the value of rapid relaxation (U _r ), equal to the decrease in the retracted fragment after 0.1 s after the termination of absorption; the value of maximum deformation (U _f ) equal to the length of the retracted fragment before depressurization. The deformation parameters were measured in the fifth suction / recovery cycle. Irradiation with ultraviolet light was carried out 3 times a week for 6 weeks using a Toshiba SE UV lamp located at a distance of 40 cm from the irradiated skin of animals for 3 minutes. For irradiation, as well as intradermal injection, animals were fixed in special machines. Mesotherapy preparations were administered weekly for 6 weeks intradermally by four injections of 2.5 μl uniformly in the calcaneal region. To control the effect of manipulations, we used a group of animals that were injected with saline. The introduction of drugs was carried out per day (Tue.) Without the UV irradiation procedure (Mon, Wed, Fri). The elastic properties of the skin were measured three days after the last UV irradiation procedure.

The effect of mesotherapeutic procedures using PERFTORAN ^® emulsion and Embryoblaste preparation on accelerated aging of rat skin under ultraviolet radiation are shown in Table 4.

According to the data obtained, the effectiveness of mesotherapy using the PERFTORAN ^® perfluorocarbon emulsion is not only not inferior to the effectiveness of Embryoblaste widely used in cosmetology practice, but even surpasses it. So the use of both drugs reliably shifted the skin parameters of the irradiated skin towards the values of intact skin. However, in the case of Embryoblaste, the values of the individual parameters of elasticity (U _f and U _r ) still significantly differed from those in intact animals.

In general, the above examples demonstrate the effectiveness of the use of perfluorocarbons as an effective tool in cosmetic mesotherapy procedures. Injections of the proposed drugs are carried out at minimum intervals of 7-14 days, with maintenance therapy - 1 time in 1-6 months. In general, the basic principle is as follows: the next procedure begins only when the previous action has ended.

Table 1. Types of Mesotherapy Tools Classical Mesotherapy Special needles
Syringes
Mesoinjectors
Pathogenetic drugs Carboxytherapy (Carbon Dioxide) Special needles Oxytherapy (administration of oxygen under pressure) Special Dispensers
Devices that administer drugs under oxygen pressure (oxygen mesotherapy) Needleless Mesotherapy Electrophoresis
Phonophoresis
Laser photomesotherapy

Table 2. Name The basis Act Cofinet Caffeine Lipolitic Siliconoro Organic silicon Connective tissue regenerant Phosphol Phosphatidylcholine Membrane Protector Structum Chondroitin sulfate Connective tissue regenerant Gotu cola Centella Asiatica Extract Collagen synthesis stimulator Chrysantehellum Chrysanthemum extract Lipolitic

Table 3. No. p / p Name The content of the component in 100 ml of emulsion Component Component function one NIZAFTEM ^® -N Perfluorodecalin 36 ml Gas transportation Perfluoropolymethyl 4 ml Gas transportation isopropyl ether
Oxide block copolymer 5.0 g Emulsifier, stabilizer ethylene and propylene (Poloxamerl88) 2.6 g Osmotic agent Sorbitol Water up to 100 ml 2 NIZAFTEM ^® -L Perfluorodecalin 40 ml Gas transportation Soy phosphatidylcholine 4.0 g Emulsifier, stabilizer (Lipoid S-40) 2.6 g Osmotic agent Sorbitol Water up to 100 ml 3 PERFTORAN ^® Perfluorodecalin 6.7 ml Gas transportation Perfluoromethylcyclohexyl 3.3 ml Gas transportation pyridine 4.0 g Emulsifier, stabilizer Block copolymer of ethylene and propylene oxides 0.32 g Osmotic agent (Poloxamer 188) up to 100 ml Salt supplement Water

Table 4. Elasticity parameter intact animals The value of the parameter of elasticity (mm)
animals after irradiation and intradermal injection saline PERFTORAN ^® "Embryoblaste" U _e 0.041 ± 0.010 0.018 ± 0.010 * 0.039 ± 0.010 ^# 0.030 ± 0.010 ^# U _f 0.058 ± 0.010 0.033 ± 0.010 * 0.054 ± 0.010 ^# 0,048 ± 0,010 * ^{, #} U _r 0.040 ± 0.010 0.017 ± 0.010 * 0.035 ± 0.010 ^th 0.028 ± 0.010 * ^{, #} U _v 0.024 ± 0.010 0.038 ± 0.010 * 0.028 ± 0.010 * 0.026 ± 0.010 ^# Note. * - significantly different from intact animals, ^# - significantly different from irradiated animals that were injected with saline. Data are presented as mean ± standard deviation, n = 10 for each group.

Claims (2)

1. Means for mesotherapy, characterized in that it contains an aqueous emulsion of perfluororganic compounds at a concentration in the emulsion of 5-95 vol.%. 2. The tool according to claim 1, characterized in that the emulsion of perfluororganic compounds is direct, reverse or multiple.

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