Method for producing preparation for photodynamic therapy

FIELD: medicine, pharmaceutical, cosmetic and food industry.

SUBSTANCE: method involves destruction and hydrolysis of alga taken among the group including: laminaria, zooster and focus followed by separating the cytoplasmatic fraction and addition of food acid polyvalent metal salt. Then correction of acidity is carried out to obtain pH = 5-7 followed by drying and conversion of fraction to form useful for storage by sterilization or freezing, or drying, or preserving. The food acid polyvalent metal an aqueous solution is added preferably in the amount 0.75% of preparation mass. Invention provides expanding functional capacity of the preparation with respect to stimulation of processes in healing wounds and burns, regeneration of post-traumatic damages, oncoprotecting and photo-sensitizing activity with respect to cancer cells at maximal absorption of light radiation in the region 630-770 nm.

EFFECT: improved producing method, valuable medicinal properties of preparation.

11 cl, 3 ex

 

The invention relates to pharmaceutical, cosmetic and food industry, in particular to methods for production of biologically active drug for photodynamic therapy of marine plant materials in the form of a gel, a fluid, dry or plastic multicomponent mixture.

There is a method of production of the drug, partially suitable for photodynamic therapy, in which consistently perform preliminary degradation and hydrolysis of at least one algae with subsequent conversion to a form suitable for storage (1).

The disadvantages of this method are receiving funds from a narrow range of application and low efficiency.

There is also known a method of production of a drug for photodynamic therapy, in which consistently perform preliminary degradation and hydrolysis of algae and subsequent mechanical separation of the cytoplasmic fraction (2).

The disadvantages of this method are receiving funds from a narrow range of application, effective, mainly with thermal light burns.

An object of the invention is to create an efficient production method of a new effective drug for photodynamic therapy and the expansion of the Arsenal of methods of production of medicinal and PR is a preventive means of this assignment.

The technical result, provides solution to this problem is to expand the functionality of the final product in terms of stimulating the healing process of wounds and burns, regeneration of traumatic injuries, onkoprotektornye and fotosensibiliziruyuschimi activity against cancer cells at the maximum absorption of light radiation in the region of 630÷770 nm.

The essence of the invention is that the method of production of a drug for photodynamic therapy, in which successively perform the decomposition and hydrolysis of at least one algae from the group including kelp, toaster and fucus, with subsequent mechanical separation of the cytoplasmic fraction and the addition of an aqueous solution of salt of polyvalent metal food acids, and then correction of acidity to obtain a pH of 5-7, drying (water distillation) and transform into a form suitable for storage.

Preferably add water 5-25% solution of salt of polyvalent metal food acid in the amount of 0.75% by weight of the drug, which is converted into a form suitable for storage by sterilization or freezing or adding preservatives or drying, and the destruction of algae to produce a homogeneous condition by homogenization by grinding, or by createstr cciii at temperatures below minus 4° With, or by cavitation processing at the speed of the expiration of not more than 120 m/s, and the hydrolysis is carried out in an acidic environment with a pH<5, or in an alkaline environment with a pH>7.

Kelp use sugar or Japanese and cytoplasmic fractions produced within 6-8 hours after completion of the hydrolysis filtration method at a temperature of 20-50°, discarding the precipitate, or by centrifuging at 4000-14000 rpm for 10 min at a temperature of 20-50°, discarding the precipitate.

As a solution of salt of polyvalent metal using a solution of a metal salt from the group of trace elements or groups of macronutrients, are part of the human body, for example as a solution of salt of polyvalent metal use a solution of salt from the group of: calcium chloride, Asparaginate magnesium, calcium malate, zinc tartrate, citrate of iron, calcium citrate, magnesium carbonate, aluminum pyrophosphate, pyrophosphate chromium, and the solution of salt of polyvalent metal is added in amounts selected on the basis of metal content in the finished product not more than 7 mg/kg

Use subject to degradation, hydrolysis and separation of extracellular and intracellular fluid seaweed and salt, polyvalent metal food acids allows to obtain new and effective drug with a maximum absorption of radiation by the region 630-770 nm.

The principle of action of the drug following.

Drug testing terrie is administered intravenously or topically. Active ingredients of the drug photodynamic therapy - chlorines (cyclic tetrapyrrole chlorin nature), mainly chlorin e6accumulate in damaged tissue, namely selectively accumulate in tumors by binding structures, excessive in tumors compared with healthy tissues. Accumulation occurs within from 3 hours to 3 days, during which the patient must comply with the “dark mode”. The presence of cations of polyvalent metal reduces the time required compliance with “dark mode” almost 20%. Low-intensity laser radiation or radiation other incoherent light source is focused on the affected area. The depth of penetration into the tumor and the effectiveness of treatment depends on the wavelength. The closer the wavelength of the light used to activate the drug photodynamic therapy to the wavelength of its absorption maximum, in this case 630-770 nm, the deeper it can penetrate light into the tissue and cause maximum damage to the deep-lying layers of the tumor. Drug molecules photodynamic therapy in the irradiation process, start to intensively absorb the light energy. Excited by light product photodynamic is what therapy is very active and reacts with oxygen, producing a “singlet” atomic oxygen, which has cytotoxic properties and calling the active oxidation processes (exceeding a threshold) in diseased cells. I.e. the drug photodynamic therapy comes into an active state, releasing toxic compounds that damage blood vessels that feed tumor cells, leading to necrosis, destruction and death of diseased cells. Developing a strong swelling of damaged tissues and their subsequent death and removal from the body of dead cells. For each drug photodynamic therapy uses a light source with a specific wavelength corresponding to the peak (maximum) absorption. To internal organs radiation supplied by the fiber-optic catheters.

Flowing the process is not accompanied by thermal tissue damage, and, in comparison with the methods of coagulation and laser surgery, do not damage healthy tissue due to its selectivity, and can be used in outpatient, and even in those cases where surgical treatment is contraindicated.

Dose constituting 10% of therapeutic drug can be used as a marker for early diagnosis of diseases.

The functionality of the final product allow you to use it also to stimulate the process C is gillenia wounds and burns, regeneration of posttraumatic injuries as intercellular and intracellular fluid, modified by cations of polyvalent metal, produces the stitching deformed and destroyed in the damage zone of organic molecules in the skin and other organs, stimulates the process of tissue respiration and circulation in this area. These properties of the drug are important in combination with onkoprotektornye and fotosensibiliziruyuschimi activity, since both the destruction and removal of malignant cells stimulated intensive recovery and healing of surrounding tissue, and the effective absorption of the additional input of valuable substances, such as vitamins, micro - and macro-elements.

The method is illustrated by the following examples.

Example 1.

Consistently perform a decomposition of algae kelp and bladder wrack, taken in a mass ratio of 1:1 by homogenization by grinding and processing in the mixer until a uniform homogeneous gelatinous consistency that does not contain coarse particles, and the hydrolysis in an alkaline environment with a pH>7. After 6 hours after completion of the hydrolysis mechanically separated by filtration method at a temperature of 32±3°zitoplazmaticescuu faction, discarding the precipitate, and add a 16% aqueous solution of aluminum pyrophosphate, which is the salt of prevalentemente food acids. Moreover, the solution of aluminum pyrophosphate added in amounts selected on the basis of metal content in the finished product not more than 7 mg/kg, in this case in the amount of 0.85% by weight of the total amount referred to input components, which is determined by calculation from the results of laboratory analyses for metals in each of the components of the mixture. Then correction of acidity to obtain a pH of 5-7, drying (water distillation) and transform into a form suitable for storage by sterilization.

To verify the validity of this preparation courses PDT 77 ill. Of these, 54 patients received one course of PDT, 2 course 12, more than 2 courses - other. The average dose was 0.7 mg/kg Used solid-state lasers “pole-2”, the energy density at the outer irradiation was 170-220 j/cm2when the irradiation of the internal organs 70-270 j/cm2with a maximum light emission in the field 670-770 nm. Complete resorption achieved in 62 patients in the other partial.

Example 2.

Initially, carry out the destruction of the algae laminaria and toaster, taken in a mass ratio of 2:3 by cavitation processing, which is performed when the flow around a cone cavitator flow blockage at the speed of the expiration of 75 m/s In plant mass generated nestec the lines of cavitation cavities. Moving in the flow and breaking up (fractions), cavities form in the zone of their collapse pulsating field of cavitation bubbles, the impact of which provides a uniform consistency, not containing coarse particles. It is followed by hydrolysis in an acidic environment with a pH<5. 8 hours after completion of the hydrolysis mechanically separated by centrifuging at 4500±250 rpm/min for 10 min and a temperature of 35±3°cytoplasmic fraction, discarding the precipitate, and then add a 15% aqueous solution of magnesium carbonate, a salt of polyvalent metal food carboxylic acid, sterilized and capsulebuy. Moreover, the solution of magnesium carbonate added in amounts selected on the basis of metal content in the finished product not more than 7 mg/kg, in this case in the amount of 1.0% by weight of the total amount referred to input components, which is determined by calculation from the results of laboratory analyses for metals in each of the components of the mixture.

Then correction of acidity to obtain a pH of 5-7, drying (water distillation) and transform into a form suitable for storage by freezing.

To verify the validity of this preparation courses 51 PDT patients. Of these 39 patients received one course of PDT, 2 course 9, more than 2 courses - other. With ednea dose was 0.75 mg/kg Used solid-state lasers “pole-2”, the energy density at the outer irradiation was 170-220 j/cm2when the irradiation of the internal organs 70-270 j/cm2with a maximum light emission in the field 670-770 nm. Complete resorption achieved in 44 patients in the other partial.

Example 3.

Consistently perform a decomposition of the algae toaster by cryodestruction, for example, due to repeated freezing-thawing at a temperature of minus 16°characterized by complex morphological changes in the cells of plant material, and hydrolysis in an alkaline environment with a pH>7. 7.5 hours after completion of the hydrolysis mechanically separated by centrifuging at 5500±250 rpm/min for 10 min and a temperature of 37±2°cytoplasmic fraction, discarding the precipitate, then add 18% aqueous solution of zinc tartrate, which is a salt of polyvalent metal food tartaric acid. Moreover, the solution of tartrate of zinc added in amounts selected on the basis of metal content in the finished product not more than 7 mg/kg, in this case in the amount of 0.9% wholesale mass of the total amount referred to input components, which is determined by calculation from the results of laboratory analyses for metals in each of the components of the mixture. Then spend a correction is acidity to obtain pH=5-7, drying (water distillation) and transform into a form suitable for storage by the addition of a preservative (formalin).

To verify the validity of this preparation courses 43 PDT patients. Of these, 32 patients received one course of PDT, 2 course 7, more than 2 courses - other. The average dose was 0.7 mg/kg of the Used diode lasers “Milon”, the energy density at the outer irradiation was 180-240 j/cm2when the irradiation of the internal organs 70-275 j/cm2with a maximum light emission in the field of 650-730 nm. Complete resorption was achieved in 30 patients in the other partial.

A specific number of components for examples 1-3 depending on the size and technical support of production, however, the technical result to be obtained by manual trial (small party) and mechanized mass production, is the same.

The drug is weakly toxic and free balance quickly excreted from the body.

Such a synergistic action of the components can improve as photodynamic effect and the healing of healthy tissue and the absorption of valuable substances, such as vitamins, micro and macro.

In the invention in an effective way preparation for photodynamic therapy and expanded the Arsenal of methods for the production of therapeutic and treatment is but prophylactic drugs for this purpose.

This expanded functionality of the final product in terms of stimulating the healing process of wounds and burns, regeneration of traumatic injuries, onkoprotektornye and fotosensibiliziruyuschimi activity against cancer cells at the maximum absorption of light radiation in the region of 630-770 nm.

Sources of information:

1. RU # 2152737, 2000

2. RU # 2174404, 2001 (prototype)

1. Method of production of a drug for photodynamic therapy, in which successively perform the decomposition and hydrolysis of at least one algae from the group comprising: kelp, toaster and fucus, with subsequent mechanical separation of the cytoplasmic fraction and the addition of an aqueous solution of salt of polyvalent metal food acids, and then correction of acidity to obtain pH 5÷7, drying (water distillation) and transform into a form suitable for storage by sterilization or freezing, or drying, or add preservative.

2. The method according to claim 1, characterized in that type of water 5÷25%solution of salts of polyvalent metal food grade acid in an amount of not less than 0.75% by weight of the drug.

3. The method according to any one of claims 1, 2, characterized in that the degradation of algae to produce a homogeneous condition by homogenization by grinding, or by cryosurgery at t is mperature below minus 4° With, or by cavitation processing at the speed of the expiration of not more than 120 m/S.

4. The method according to any one of claims 1, 2, characterized in that the hydrolysis is carried out in an acidic environment with a pH<5.

5. The method according to any one of claims 1, 2, characterized in that the hydrolysis is carried out in an alkaline medium with a pH>7.

6. The method according to any one of claims 1, 2, characterized in that the separation of the cytoplasmic fraction produced after 6-8 h after completion of the hydrolysis filtration method at a temperature of 20÷50°, discarding the precipitate.

7. The method according to any one of claims 1, 2, characterized in that the separation of the cytoplasmic fraction produced after 6÷8 h after hydrolysis by centrifuging at 4000÷14000 rpm for 10 min at a temperature of 20÷50°, discarding the precipitate.

8. The method according to any one of claims 1, 2, characterized in that use kelp sugary or Japanese.

9. The method according to any one of claims 1, 2, characterized in that as a solution of salt of polyvalent metal using a solution of a metal salt from the group of trace elements or groups of macronutrients, are part of the human body.

10. The method according to any one of claims 1, 2, characterized in that as a solution of salt of polyvalent metal use a solution of salt from the group of: calcium chloride, Asparaginate magnesium, calcium malate, zinc tartrate, citrate of iron, calcium citrate, to rbonate magnesium, the pyrophosphate aluminum pyrophosphate chromium.

11. The method according to any one of claims 1, 2, characterized in that the solution of salt of polyvalent metal is added in amounts selected on the basis of metal content in the finished product not more than 7 mg/kg



 

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