Bioregulatory complex possessing tissue-specific regenerative action, method for preparing it and method of treating cataract using it. RU patent 2513994.

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a bioregulatory complex possessing regenerative action. The bioregulatory complex possessing regenerative action prepared of gills of post-settlement Atlantic salmon (Salmo salar L.) activated for a prolonged life cycle with symbiotic pearl shell (Margaritifera margaritifera) alevins, containing peptides and oligosaccharides with certain physicochemical characteristics. A method for preparing a bioregulatory complex consisting the fact that the gills are prepared in the Atlantic salmon fished out after the settlement and placed into water or ethanol; the prepared gill extract is filtered; monoatomic aliphatic alcohol is added to the filtrate that is kept to deposit any foreign matters; the deposition is filtered off; the filtrate is evaporated dry, dissolved in water and cleaned in the specific environment. A method of treating early cataract.

EFFECT: bioregulatory complex has an effective inhibitory action on developing cataract and possesses an evident regenerative effect.

6 cl, 1 dwg, 1 tbl, 6 ex

The invention relates to a method of obtaining biologically active complex, which can find application in medicine, pharmaceutics as active substances in the production of drugs that stimulate physiological and reparative regeneration, in treatment diseases and traumatic lesions of the skin, as well as wound healing products (with wounds of different etiology: ulcers, burns, secondarily infected).

Recently in Arctic ecosystems found a unique phenomenon, when the biochemical program of aging and post-reproduction suicide Atlantic salmon (salmon) can be turned off under the influence of symbiotic organism - periodic fabric of the parasite epithelium gills salmon - larvae of freshwater pearl mussels. Thereby limiting the lifetime of salmon is extended to 13 years, and salmon instead of single spawning became able to multiply many times - up to 5-6 times. Larva pearl neutralizes senile changes in the regulatory system salmon "hypothalamus - pituitary - peripheral endocrine gland, the hypothalamus". Larvae of the mollusc through the blood constantly supply the nerve cells "owners" natural antidepressant and probably neurotransmitters or their predecessors. Preliminary analysis has shown that the parasite-symbiont secretes in the blood of the host of water-soluble substances, including amino acids, peptides and glycoproteins (Zyuganov V.V. (2005). Survivor-parasite life-prolonging master - Oyster Margaritifera margaritifera disables the program of accelerated aging in salmon Salmo salae [Zyuganov V.V. (2005), DAN. t, №5, s-705].

In this regard, the secret gills Atlantic salmon, activated on extended life cycle symbiotic mollusc larvae-oyster can serve as a source of biologically active complex, which can find application in medicine in the creation of new medicines.

Known biologically active complex "Arctic", used to treat cancer [RF patent №2324489, published. 2008]. This complex is a mixture of crude complex extract secret gills Atlantic salmon and the secret of the renal tubules spawning male stickleback alcohol tincture-elixir (high-quality five-year cognac extracts). The method of obtaining biologically active complex is not disclosed.

Cataract is one of the most common causes of vision loss, and the number of patients with age complicated and traumatic cataracts increases with every year. Currently in the treatment of cataracts is dominated by radical surgical method (Kopaev V.G. Eye diseases. The textbook. M: Medicine, 2002, C-268). Therapeutic treatment depart on the second plan. Therefore, the search for drugs that provide acceptable therapeutic effect.

The closest to the proposed method of treatment of the initial cataract is a method of treatment with the drug "Valentin" (RF patent №2367387, published. 2009), namely, that neutral solution low-molecular fractions of peptides derived from the faction acidic protein tissue extracts lenses vertebrates, highlighted by fractionation 100%solution of ammonium sulfate ("Valentin"), instilliruut in conjunctival cavity 9, 12 and 17 h 2 drops in both eyes 2 times with an interval a 5 min a course of 2-6 months with an interval of 10 days and every 30 days.

However, this treatment has a significant disadvantage in a significant seal back capsule of the eye during cataract treatment the drug " Valentin" (FGI IRI GB them. Helmholtz). Data Doppler were not comparable with the clinical results and testified to the fact that the parameters of the indicators after the treatment ($180- $ 185) were higher than before the treatment ($165- $ 172) [Verigo E.N. and other Prospects of application of the preparation "Valentin" in the primary form of cataracts. Collection of conference "Nanotechnologies in the diagnosis and treatment of pathology of organs of vision". Moscow, 2008, p.34-37]

The objective of the invention is to obtain high-purity of physiologically active complex bioregulators of gills posledeistvie Atlantic salmon (Salmo salar L.), activated symbiotic larvae (glochidiata) mollusk pearl (Margaritifera margaritifera), extended life cycle.

The problem is solved bioregulatory complex with reparative tissue-specific action, received from the gills posledeistvie Atlantic salmon (SalmoSalar. L.), activated on extended life cycle symbiotic mollusc larvae pearl (Margaritifera margaritifera), containing peptides and carbohydrate content of carbohydrates in relation to the peptides 5-18% having the size of particles 80-220 nm 0.1%aqueous solution at a temperature of 20 C and UV range with maximum absorption at 255-270 nm, "shoulder" when 200-229 nm and the ratio of the intensity of absorption at 210 nm to the absorption intensity at 255 nm equal to 4,5-5,5, and the following characteristics for 0.1%aqueous solution: isoelectric point of 3.8 4.4 and Zeta potential of 30 to 40 mV at pH 6.5 and 1-3 mV at pH 4,2.

Bioregulatory complex has an inhibitory effect on the development of a cataract on the cultures eyes rats, resulting in the violation of the basic enzyme systems of the lens - lipid peroxidation and Ca 2+ -dependent protease. Bioregulatory complex significantly improves visual acuity patients with age and traumatic cataracts.

The problem is solved by the method of obtaining bioregulating complex of gills posledeistvie Atlantic salmon (Salmo salar L.), activated on extended life cycle symbiotic mollusc larvae pearl (Margaritifera margaritifera), namely, that have caught after spawning Atlantic salmon dissect the gills and put them in water or in aqueous solution 10-40% ethanol, the obtained extract gills filter, add in the filtrate monatomic aliphatic alcohol at a ratio of leachate and alcohol 3/1 up to 1/3 and leave before the deposition of impurity substances that fell sucked off, the filtrate is evaporated to dryness, dissolved in water and purified by column chromatography.

At the same time as saturated aliphatic alcohol preferably use ethanol, or propyl alcohol or isopropanol.

The problem can be solved by method of treatment of the initial cataract, namely, that introduced the aqueous solution chromatographically purified fraction of bioregulatory complex, retrieved from gills posledeistvie Atlantic salmon (Salmo salar L.), activated on extended life cycle symbiotic mollusc larvae pearl (Margaritifera margaritifera) with concentration of 10 -8 -10 -14 mg/ml in both eyes into the conjunctival cavity in the effective number three times a day course duration 2-5 months with an interval of 5-10 days every 30 days.

The solution is injected by putting every once in two steps 1-2 drops in each eye every 5-7 minutes or by spray irrigation two injection doses by 30-70 ml.

The method of obtaining of bioregulatory complex is as follows. We caught after spawning Atlantic salmon dissect the gills, containing globigii, and put them in a vessel with water or a water solution 10-40% (preferably 30%) of ethanol in the tenfold excess in relation to the mass of gills. The extract obtained gills filtered. The filtrate is placed in the chemical glass and add a monatomic alcohol (methanol, ethanol, propyl alcohol or isopropanol) at a ratio 3/1-1/3 and left overnight at +4 C. the Resulting sludge is separated from the supernatant liquid filtration. The filtrate is evaporated on a rotary evaporator to dryness and dissolve in the water. For obtaining biologically active fraction of bioregulatory complex further cleaning is performed by HPLC in facing the phases in the manual mode on the speakers type: C8; C16; C18 or C32 with a particle size of stationary phase 1.5 to 10 microns. Elution of substances with speakers Bond C18 (250x22 mm; 5 microns) are within 20 minutes by water, and then within 20 minutes 40%ethanol at the speed of the mobile phase 5 ml/min and detect peaks at 254 nm. The column applied to 2 ml of the solution.

To determine the biological activity received chromatographic fractions used adhesiolysis method [Yamskova VP, M. Reznikov Journal of General Biology, 1991, T, №2, s-191]. Faction possessing biological activity, accumulate, take in a round-bottom flask and evaporated to dryness in a rotary evaporator. Dry residue is dissolved in water, filtered and define the following physical and chemical properties:

the concentration of peptides (bicinchoninic method; Smithet.al, 1985), oligosaccharides (astronomy method; Koehler, 1952), remove the UV range and determine the particle size and Zeta potential of bioregulatory complex (dynamic light scattering).

It is established that the set of bioregulators has the following physical-chemical features:

1. The complex consists of:

A) peptides

B) oligosaccharides.

The relative content of oligosaccharides a peptide is 5-18%.

2. The average particle size of the complex for 0,1% water solution at a temperature of 20 C 80-220 nm

3. UV spectrum of the complex:

A) the maximum absorption at 255-270 nm,

B) the shoulder when 200-229 nm,

B) the ratio of the absorption intensity at 210 nm to the intensity at 255 nm is 4,5-5,5

4. Isoelectric point for 0,1% water solution of the complex - 3,8-4,4

5. Zeta potential for 0,1% water solution of the complex:

A) at a pH of 6.5 equal to 30-40 MB

B) at pH equal to 4.2 - 1-3 mW.

Examples of obtaining of bioregulatory complex.

Example 1. From caught by industrial way after spawning Atlantic salmon analyzed gills, containing globigii, and put them in a vessel with water solution of 30% of ethanol in the tenfold excess in relation to the gills. Received the drug is transported in such kind of places catching salmon. Alcoholic extract gills was filtered through a funnel off, equipped with a filter paper, in the suction flask under vacuum, water-jet pump. The filtrate was placed in chemical glass and added ethanol at a ratio of 1/3 and left overnight in a refrigerator at +4 C for deposition of impurity substances. The formed sediment was separated from the supernatant fluid filtration through a funnel off, equipped with a filter paper, in the suction flask under vacuum, water-jet pump. The filtered solution is evaporated on a rotary evaporator to dryness and dissolved in water. For obtaining biologically active fraction of bioregulators further purification was performed by column chromatography (HPLC) on the speaker Bond C18 (250x22 mm; 5 mm). Within 20 minutes elution substances carried by water, and then within 20 minutes 40% ethanol at the speed of the mobile phase to 5 ml/min and detect peaks at 254 nm. The column inflicted 2 ml.

Faction possessing biological activity, accumulated, moved in a round-bottom flask, and was evaporated to dryness in a rotary evaporator. Dry residue is dissolved in water, filtered and identified the following physical and chemical properties:

1. The complex consists of:

The relative content of oligosaccharides to the peptides was 10%.

2. The average particle size of the complex for 0,1% water solution at a temperature of 20 C to 150 nm

3. UV spectrum of the complex:

A) the maximum absorption at 270 nm,

B) the shoulder when 200-225 nm,

B) the ratio of the absorption intensity at 210 nm to the intensity at 270 nm is 4,5.

4. Isoelectric point for 0,1% water solution of the complex of 4.1

5. Zeta potential for 0,1% water solution of the complex:

A) at a pH of 6.5 equal to 40 mV

B) at pH equal to 4.2 - 2 mV.

Example 2. From caught by industrial way after spawning Atlantic salmon analyzed gills, containing globigii, and put them in a vessel with water solution of 10% ethanol in a fivefold excess in relation to the gills. The drug was taken in this form from the place of catching salmon. Alcoholic extract gills was filtered through a funnel off, equipped with a filter paper, in the suction flask under vacuum, water-jet pump. The filtrate was placed in chemical glass and added ethanol at a ratio 3/1 and left overnight in a refrigerator at +4 C for deposition of impurity substances. The formed sediment was separated from the supernatant fluid filtration through a funnel off, equipped with a filter paper, in the suction flask under vacuum, water-jet pump. The filtered solution is evaporated on a rotary evaporator to dryness and dissolved in water. For obtaining biologically active fraction of bioregulators further purification was performed by column chromatography (HPLC) on the speaker Ultrasphere C8 (250x22 mm; 10 microns). Within 25 minutes elution substances carried by water, and then within 35 minutes 40%ethanol at the speed of the mobile phase 4 ml/min and detect peaks at 254 nm. The column inflicted 2 ml.

Faction possessing biological activity, accumulated, moved in a round-bottom flask, and was evaporated to dryness in a rotary evaporator. Dry residue is dissolved in water, filtered and identified the following physical and chemical properties:

1. The complex consists of:

The relative content of oligosaccharides a peptide is 15%.

2. The average particle size of the complex for 0,1% water solution at a temperature of 20 C to 220 nm

3. UV spectrum of the complex:

A) the maximum absorption at 260 nm,

B) the shoulder when 200-229 nm,

B) the ratio of the absorption intensity at 210 nm to the intensity at 265 nm equal to 5.3.

4. Isoelectric point for 0,1% water solution of the complex - 5,2

5. Zeta potential for 0,1% water solution of the complex:

A) at a pH of 6.5 equal to 30 mV

B) at pH equal to 4.2 - 3 mV.

The concentration of peptide component of bioregulatory complex in solution was set equal to 0.1 mg/ml by dilution with water (qualification Milli-Q). Then by serial dilution with water received solutions with a concentration of 10 -8 to ...10 -14 mg/ml necessary for biological testing.

Model cataractogenesis on the cultures of the lens Wistar rats

Use the extracted the lens breed rats Wistar.

Lenses allocated microsurgical in sterile conditions from the eyes of the animals under the control of binocular microscope. All animals previously had narcotizirutego.

Aqueous solution of bioregulatory complex (c=10 -10 mg/ml) were added into the medium in the early cultivation of crystals. All vials prepared cultures hermetically closed.

For cultivation of lenses used the medium 199, containing 80 mg/l gentamicin sulfate. The lenses were placed in glass Buxi in 5 ml of environment and cultivated at a temperature of 37 C for 4-5 days. On the third day produced a change of culture medium.

Lenses rats were divided into three groups, 4-5 lenses each:

1-I control group - for the cultures of 4.5 ml of culture medium 199 added 0.5 ml of physiological solution;

2-I control group - for the cultures of 4.5 ml of culture medium 199 added 0.5 ml damaging agent concentration: hydrogen peroxide 0.5 mm or calcium chloride 15 mm, respectively;

3-I experienced group - to 4,0 ml of culture medium 199 added 0.5 ml damaging agent in the same concentration as in the 2-nd group, and 0.5 ml of the solution faction bioregulatory complex.

After 5 days of cultivation lenses were removed and studied spectrophotometrically. The experiences of each experimental series was repeated three times.

Determination of the degree of cataract rat by spectrophotometry

The degree of cataract studied spectrophotometrically at the wavelength 340, 405, 490, 550, 630 nm. Lenses rats were placed in 24-well plate and in the absence of the aqueous phase was determined by the intensity of light scattering at these wavelengths.

In all experiments the lens animals remained transparent during the whole time of cultivation in control vials (in the environment of cultivation, without adding damaging agents).

Model cataractogenesis on the cultures of the lens rats in vitro

In the study of the impact bioregulatory complex (BC) on the development of cataractogenesis in the lens of the eye rats in vitro had the following series of experiments:

№1 - nutritional Wednesday 199;

№2 - nutritional Wednesday 199 containing H 2 O 2 ;

№3 - nutritional Wednesday 199 containing H 2 O 2 +solution Bq;

№4 - nutritional Wednesday 199 containing CaCl 2 ;

№5 - nutritional Wednesday 199 containing CaCl 2 +solution BC.

Figure 1 shows the results of spectrophotometric study of the effect of bioregulatory complex on cataractogenic in the lens rats induced by the influence of hydrogen peroxide and calcium chloride. The abscissa axis shows the number of series of experiments on the axis of ordinates the optical density. Right - data wavelengths (nm). In each series of experiments shows the optical density for wavelengths (from left to right): 630, 550, 490, 405 and 340 nm.

Data series №2, №3 (control-experience) and №4, №5 (control-experience) authentic p<0.05.

Presented in figure 1, the results suggest that the addition in the cultural medium Bq helps to increase transparency, lenses rats approximately in two times in comparison with lenses that are cultured in medium containing cataractogenic agents.

Thus, this study is conducted on the cultures of the lens rats, it was established that the application of bioregulatory complex prevents the development of cataractogenesis animals in vitro.

In result of studies it is established that bioregulatory complex has an inhibitory effect on the development of cataracts, resulting in the violation of the basic enzyme systems of the lens - lipid peroxidation and Ca 2+ -dependent protease.

Study of the activity of bioregulatory complex in a stimulating effect on the regeneration processes in the skin of rats

Burns in rats caused on the back of boiling water for 20 seconds. Was applied standard of burn wound II-III A degree with total area of 6 cm2 of skin and soft tissues of the back with the preliminary removal of hair.

In control (the wound was not processed) on the 14th day after burn watched almost complete epithelization, scab tightly adjoined to the skin. Observed a slight inflammation in the dermis and the muscle layer. Was necrosis hair follicles. In the muscle layer was observed cavity between the fibers, formed as a result of inflammation. Under the epithelium were visible accumulation of neutrophils. Granulation tissue fiber-cell.

To 25 days was formed connective tissue scar burn with insignificant presence of neutrophils is observed necrosis hair follicles. Strup practices completely detached.

On 25 day there was a complete epithelization with the expressed multicellular epithelium, the almost complete detachment of scab. In the dermis inflammation practically not observed. Formed cellular-fibrous granulation tissue, collagen fibres not loose. Hair follicles in the area of the injury was not observed.

Thus, the proposed bioregulatory the complex has a specific pharmacological activity in the form of a stimulating influence on the process of reparative regeneration of the skin, manifested in the healing of burn wounds, relieve inflammation and stimulating the proliferation of the epithelium of the skin in the area of damage and dermal fibroblasts.

In the wounds with full epithelization epithelium over the burn area was significantly greater thickness compared located near the skin.

Conclusion: high efficiency of the offered complex in the healing of burn wounds III a degree, fast removal of inflammation and stimulates the proliferation of the epithelium of the skin in the area of damage and dermal fibroblasts.

The proposed method of cataract treatment carried out as follows.

The concentration of peptide component of bioregulatory complex in solution was set equal to 0.1 mg/ml by dilution with water (qualification Milli-Q). Then by serial dilution with water received solutions with the correct concentration and added CaCl 2 to the concentration of 0.1 mg/ml (drug "Anticataract").

Installed into the conjunctival cavity in neutral solution bioregulatory complex in doses (10 -8 -10 -14 ) mg/ml Instillation was carried out in 9, 13 and 18 hours 2 drops or spray in both eyes 2 times with an interval of 5-7 min rate duration of 2-5 months with an interval of 10 days and every 30 days.

Clinical studies for the treatment of cataract eye drops on the basis of bioregulatory complex

The drops: water sterile solution of bioregulatory complex (c=10 -10 mg/ml)containing 0.1 mg/ml CaCl 2 ("Anticataract").

I group (37 volunteers - 37 eyes) with the initial age-related cataracts, and in the absence of opacities in the lens.

I. the group is divided into subgroups:

1. Beredskapslagrade cataract (9 patients - 18 eye), age 40-59 years. GOATS from 0.4 to 0.8.

2. Cortical cataract (8 patients - 16 eyes), age 48-67 years, GOATS from 0.6 to 0.9.

3. Zadnekamernye cataract (20 patients 40 eyes) age 55-72 year, GOATS from 0.2 to 0.8.

II group (8 patients - 6 eyes) with traumatic cataracts.

Research methods:

1. General clinical research methods.

2. Biomicroscopic the photography.

3. The fundus photography.

4. ULTRASONIC dopplerography.

In the I-th group used eye drops or spray.

Drops installed in co bag eye of 2 drops 2 times with an interval of 5-7 minutes at 9-00, 13-00, 18.00 p.m., with a break for 10 days and every 30 days for 2-5 months.

The spray was watered the cornea of the eye 2-3 injection 2 times with an interval of 5-10 minutes, in the same mode that drops.

Held 4 times a dynamic survey (external inspection, GOATS (korrigirovanija visual acuity), perimetry, samples Schirmer and burrowing.)

The results of the studies on volunteers following.

Dynamics of visual acuity in both groups are given in table. Visual acuity improved. By age cataract "Front Capsule-7": after treatment with eye drops and spray "Anticataract" GOATS (korrigirovanija visual acuity) improved by 3 positions, to ensure visual acuity is not korrelirovalisj.

"Cortical layers-7": GOATS after treatment equal to 7 (vision improved on two lines of the table).

"The posterior capsule-19": Age-related cataract: visual acuity improved by two positions. GOATS before treatment 5, after treatment 7. (Table)

Traumatic cataract: visual acuity improved by one position in the table. GOATS before treatment 5, after treatment 6. (Table)

Dynamics of visual acuity, depending on the location and etiology of cataract

Etiology of cataract

Localization of turbidity

Before treatment

After treatment

Age (33 patients)

Front capsule (7)

Cortical layer (7)

The posterior capsule (19)

Traumatic cataract (7 patients)

Examples of cataract treatment

Example 1.

The patient Century 32 years. The diagnosis of traumatic cataracts with intraocular foreign body (a complicated cataract), the right eye vis=0,1 n/K.

Assigned to treatment with the drug "Anticataract" according to the following scheme: the drug is installed in into the conjunctival cavity 9, 13 and 18 hours 2 drops with an interval of 5-7 minutes A course of 3 to 4 months with a break for 10 days and every 30 days.

Three months revealed a decrease in turbidity in the cortical layer of the crystalline lens, improved visualization parts of the fundus (according to the photographic recording on the fundus-camera).

Visual acuity after treatment:

Right eye vis=0.6 n/a

Thus, at use of the drug "Anticataract" patients showed a significant improvement of subjective and objective indicators of view, indicating partial resorption of cataract and improving the function of the lens.

Example 2.

The patient is 27 years old was injured left eye jumped knot at the sawmill. After 2 months have seen a dramatic reduction of vision. Diagnosis: initial traumatic cataract in his left eye.

Indices before treatment:

Right eye=0,3 from SF. -1,5=0,9-1,0.

Left eye=0,1 with SF. -2,0=0,7

Clinically in the left eye detected traumatic midriaz, opacities in front of cortical layers and front lens capsule. The patient the drug treatment "Anticataract" in the left eye as follows: the drug is installed in into the conjunctival cavity 9, 13 and 18 hours 2 drops with an interval of 5-7 min Rate from 3 to 4 months with a break for 10 days and every 30 days.

Parameters after treatment:

Left eye=0,2 with SF. -2.0=0,9-1,0.

After treatment, subjectively the patient marks the improvement of view, the disappearance of the "fog" in front of the eye. Visual acuity of the left eye correction=0,9-1,0.

Example 3.

The patient Century 67 years. Diagnosis: Primary senile cataract. Complaints about reduction of view in the last 9 years, buried "Taufon", no effect was observed.

Visual acuity before treatment vis=0,2c+3,0=0,5.

The patient the drug treatment "Anticataract" in both eyes as follows: the drug is installed in into the conjunctival cavity 9, 13 and 18 hours 2 drops with an interval of 5-7 min rate from 3 to 4 months with a break for 10 days and every 30 days.

Three months revealed a decrease in turbidity in the cortical layer of the crystalline lens (according to biomicroscopy), and the lens density (according to the data of ULTRASONIC dopplerography from $ 90 to $ 70), improved visualization of the parts of the eye.

Parameters after treatment: vis=0,4c+2,5=0,8

Thus, the proposed bioregulatory complex - drug "Anticataract" - has antikataraktnoe action, which leads to reduction of the area of the haze, the reduction of density of matter in the lens and as a result, the lens astigmatism. This drug prevents progression of cataracts, particularly traumatic Genesis, causes partial or complete resorption of opacities in the lens layers.

The study suggests that changes in the lens layers are associated with subtle biochemical processes going on the background of drug treatment "Anticataract", and require further study, particularly if it is necessary to consider localization haze and etiologic mechanisms underlying the development of cataracts.

Thus, the drug "Anticataract" in the course of treatment is not condenses back capsule of the eye and more effective than the drug Valentin".

1. Bioregulatory complex with regenerative effect derived from the gills posledeistvie Atlantic salmon (Salmo salar L.), activated on extended life cycle symbiotic mollusc larvae pearl (Margaritifera margaritifera), containing peptides and oligosaccharides with the content of carbohydrates in relation to the peptides 5-18% having the size of particles 80-220 nm 0.1%aqueous solution at a temperature of 20 C and UV spectrum with the maximum absorption at 255-270 nm, with shoulder when 200-229 nm and with the ratio of the absorption intensity at 210 nm to the absorption intensity at 265 nm equal to 4,5-5,5, and the following characteristics 0.1%aqueous solution: isoelectric point of 3.8 4.4 and Zeta potential of 30 to 40 mV at pH 6.5 and 1-3 mV at pH 4,2.

2. The method of obtaining bioregulating complex according to claim 1, which consists in the fact that we caught after spawning Atlantic salmon dissect the gills and put them in water or in aqueous solution 10-40% ethanol, the obtained extract gills filter, add in the filtrate monatomic aliphatic alcohol at a ratio of leachate and alcohol 3/1 up to 1/3 and leave before the deposition of impurity substances that fell sucked off, the filtrate is evaporated to dryness, dissolved in water and purified by column chromatography.

3. The method of claim 2, characterized in that as saturated aliphatic alcohol use ethanol, or propyl alcohol or isopropanol.

4. The treatment for the initial cataract, namely, that introduced the aqueous solution chromatographically purified fraction of bioregulatory complex according to claim 1 with concentration of 10 -8 -10 -14 mg/ml in both eyes into the conjunctival cavity in the effective number three times a day course duration 2-5 months with an interval of 5-10 days every 30 days.

5. The method according to claim 4, wherein the solution imposed by putting every once in two steps 1-2 drops in each eye every 5-7 minutes.

6. The method according to claim 4, wherein the solution imposed by spray irrigation two injection doses by 30-70 ml.