Drug preparation used for cartilage diseases

FIELD: medicine.

SUBSTANCE: invention refers to medicine and pharmacology, and represents the use of a pharmaceutically acceptable salt of alpha-ketoglutaric acid for preparing a pharmaceutical composition for treatment or prevention of cartilage diseases in response to weight loss and/or nutritional disorder, poor nutrition, gastrectomy, partial gastrectomy or gastric ligation.

EFFECT: invention provides effective relief and management of articular pain and arthritis.

20 cl, 8 tbl, 3 dwg

 

The SCOPE of the INVENTION

The present invention relates to pharmaceutical compositions and the use of such compositions for the treatment, relief and prevention of conditions associated with loss of cartilage and associated pain, or prevention of osteoarthritis and rheumatoid arthritis and associated pain.

BACKGROUND of the INVENTION

One in three adults in the industrialized world suffer from chronic pain associated with arthritis symptoms. The most common symptom - persistent pain - can appear as hip pain, knee pain, pain in the hand and pain in the wrist, as well as joint pain in other parts of the body. The symptoms cause pain and lead to economic losses, when the subject is forced to stop work or reduce the number of working hours. Large sums are spent on medical treatment. Obviously, there is a need for cost-effective solutions for the relief or treatment of joint pain and arthritis.

The INVENTION

The implementation of the invention includes the use of a substance, comprising at least one member from the group consisting of alpha-Ketoglutarate acid, glutamine, glutamic acid, and pharmaceutically acceptable salts of these acids, amides of alpha-Ketoglutarate acids and amino acids or d is - or tripeptides of glutamine and other amino acids, tripeptides of glutamine and other amino acids, dipeptides of glutamic acid and other amino acids, tripeptides of glutamic acid and other amino acids and pharmaceutically acceptable salts of these dipeptides and tripeptides, pharmaceutically acceptable physical solutions alpha Ketoglutarate acid or its pharmaceutically acceptable salts and at least one amino acid to obtain a pharmaceutical composition for the treatment or prevention of conditions inflammatory or noninflammatory lesions of the cartilage and pain associated with it.

Further implementation of the invention includes the use of the above substances for the treatment or prevention of osteoarthritis and rheumatoid arthritis and associated pain.

Further implementation of the invention includes the use of the above substances for the treatment or prevention of lesions of the articular cartilage in the States, including weight loss and/or malnutrition, or gastrectomy, partial gastrectomy or ligation of the stomach.

Further implementation of the invention includes the use of the above substances for the treatment or prevention of lesions of the articular cartilage in the States, including malnutrition.

Further implementation of the invention includes the use of the above substances for pain relief associated with what rajaniemi articular cartilage at the above conditions.

Further implementation of the invention includes the use of the above substances for the treatment or prevention of osteoporosis associated with gastrectomy.

BRIEF DESCRIPTION of DRAWINGS

The present invention will be explained in the following description with the aid of preferred implementations, case studies and accompanying drawings, in which

Figure 1 - chart describing the influence of diet alpha-Ketoglutarate and gastrectomy on the body weight of rats

Figure 2 - four transilluminated pictures of the upper section of the skull (cranial vault) of the experimental rats,

Figure 3 - chart showing the area of the grooves found on transillumination photographs of the skull vault experimental rats.

DETAILED description of the INVENTION

Thus, according to one aspect of the present invention provides a new use of at least one member of the group consisting of alpha-Ketoglutarate acid, glutamine, glutamic acid, and pharmaceutically acceptable salts of these acids, amides of alpha-Ketoglutarate acids and amino acids or di - or tripeptides of glutamine and other amino acids, tripeptides of glutamine and other amino acids, dipeptides of glutamic acid and other amino acids, tripeptides of glutamic acid and other amino acids, pharmace is almost acceptable salts of these dipeptides and tripeptides, pharmaceutically acceptable physical solutions alpha Ketoglutarate acid or its pharmaceutically acceptable salts and at least one amino acid to obtain a pharmaceutical composition for the treatment or prevention of conditions of osteoarthritis, rheumatoid arthritis and the destruction of cartilage and pain associated with the aforementioned disorders.

According to a preferred implementation of the invention applied alpha-Ketoglutarate acid, or its salts of alkaline or alkaline earth metals, or combinations of the above compounds.

According to another aspect of the present invention provides a method of treatment or prophylaxis of a condition of growing pains for at least one member of the group consisting of osteoarthritis in a mammal, including a human, where the method includes introduction to the subject for such treatment or prevention a valid pain doses of at least one member of the group consisting of alpha-Ketoglutarate acid, glutamine, glutamic acid, and pharmaceutically acceptable salts of these acids, amides of alpha-Ketoglutarate acids and amino acids or di - or tripeptides of glutamine and other amino acids, tripeptides of glutamine and other amino acids, dipeptides of glutamic acid and other amino acids, tripeptides of glutamic acid and other amino acid is t and pharmaceutically acceptable salts of these dipeptides and tripeptides, pharmaceutically acceptable physical solutions alpha Ketoglutarate acid or its pharmaceutically acceptable salts and at least one amino acid.

According to a preferred implementation of these aspects are introduced alpha Ketoglutarate acid, or its salts of alkaline or alkaline earth metals, or combinations of the above compounds. Most preferably the introduction of alpha-Ketoglutarate sodium.

The pharmaceutical preparations of the active component or components used in accordance with the present invention, may be introduced vertebrate, including mammals, such as rodents, such as mice, rats, Guinea pigs or rabbits; birds, such as turkeys, chickens or chickens and broilers and other animals free content; cows, horses, pigs, and pigs and other farm animals, dogs, cats and other domestic animals, and humans.

The introduction can be done in different ways depending on the form of the spine, which is the treatment, the condition of the vertebrate in need of named methods and specific indications for treatment.

In one implementation, the introduction is in the form of food or food additives, such as dietary supplements and/or ingredients in the form of solid food or drinks. Following the implementation of the population may be suspensions or solutions, as a drink, is described below. Also the release form can be in tablet or capsule, chewable or soluble, such as effervescent tablets and powders and other dry forms known to the skilled person skilled in the art, such as tablets, micropellet and granules.

The administration can be parenteral, rectal or oral food or food additive, as indicated above. Parenteral environment includes the solution of sodium chloride, dextrose and ringer, dextrose and sodium chloride, lactate ringer's or fixed oils.

Food and food additives can also be emulsified. The active therapeutic ingredient or ingredients may then be mixed with the shaping of the foundations, which are pharmaceutically acceptable and compatible with the active ingredient. Suitable ingredients, such as water, saline, dextrose, glycerol, ethanol or the like, and combinations thereof. Moreover, if necessary, the composition may contain minor amounts of auxiliary equipment such as moisturizing or emulsifying agents, pH buffering agents that enhance the effectiveness of the active ingredient.

You can apply various forms of parenteral food, such as solid food, liquid, lyophilized or dehydrated form. They can include solvents which do various buffers (for example, Tris-HCl, acetate, phosphate), pH and ionic enhancers, additives such as albumin or gelatin to prevent surface suction, detergents (such as tween 20, tween 80, Pluronic F68, bile salts), dissolving agents (for example, glycerin, polyethylenglycol), antioxidants (e.g. ascorbic acid, sodium metabisulfite), preservatives (e.g., thimerosal, benzyl alcohol, paraaminobenzoic acid), loosening the substance or tone modifiers (e.g., lactose, mannitol), covalent compounds with the composition of the polymers, such as polyethylene glycol the formation of complexes with metal ions, the inclusion of material in or to drugs from polymeric compositions, such as polylactic acid, polyglycolic acid, hydrogels, etc. or into liposomes, microemulsions, micella, unilamellar or multilamellar vesicles, hemolyzed erythrocytes or spheroplasts.

In one implementation of the invention, the food or food additive is introduced in the form of a drink or dry compositions of the above compounds in each of the methods according to the invention.

The drink includes an effective amount of the active ingredient or ingredients of the above compounds, together with food water-soluble carrier, such as minerals, vitamins, carbohydrates, fats and proteins. All these components are used is carried out in a dry form, if the drink is produced in a dry form. The drink is made ready for use, contains water. The finished solution can also have a certain tone and acidity, as, for example, buffer solution, according to the General instructions in the previous paragraph.

The preferred pH in the range from 2 to 5, especially from 2 to 4, in order to avoid the growth of bacteria and fungi. Sterilized drink can also be applied at a pH of about 6-8.

The drink can be used alone or in combination with one or more therapeutically effective compounds.

According to a further implementation of the invention the pharmaceutical preparation in the form of medicines for oral and rectal may be in the form of tablets, lozenges, capsules, powders, aqueous or oily suspensions, syrups, elixirs, aqueous solutions and the like, comprising the active ingredient in admixture with pharmaceutically acceptable carriers and/or additives, such as solvents, preservatives, soljubilizatory, emulsifiers, adjuvants and/or carriers used in the method and openly used in the present invention.

Used hereinafter, the phrase "pharmaceutically acceptable carriers" known qualified specialists in this field and may include, but is not limited to, 0.01 to 0.05 M phosphate buffer or 0.8% saline R is the target. In addition, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions and emulsions. Examples of nonaqueous solutions are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters, such as etiloleat. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered environment. Parenteral bases include sodium chloride, dextrose and ringer, dextrose and sodium chloride, lactate ringer's or fixed oils. Can also contain preservatives and other additives, such as, for example, antimicrobial substances, antioxidants, hepatoblastoma agents, inert gases and the like.

Amino acids that form part of amides of alpha-Ketoglutarate acid or dipeptide with glutamine or glutamic acid or tripeptides with glutamine and/or glutamic acid may be any amino acid naturally occurring in the composition of the peptides. The same refers to pharmaceutically acceptable physical solutions of alpha-Ketoglutarate acid or salts of the above compounds with at least one amino acid. Preferred amino acid/amino acid selected from the group consisting of arginine, ornithine, leucine, isoleucine and lysine.

These amino acids are preferably used in the L-configuration.

Examples of amides of alpha-Ketoglutarate acid with amino acids and di - or Tripeptide include, but are not limited to, amidon alpha Ketoglutarate acid with an amino acid selected from the group consisting of glutamine, glutamic acid, arginine, ornithine, lysine, Proline, isoleucine and leucine and amides of alpha-Ketoglutarate acid with glutamine dipeptide and any of the following substances: glutamic acid, arginine, ornithine, lysine, Proline, isoleucine and leucine and glutamic acid dipeptide and any of the following substances: arginine, ornithine, lysine, Proline, isoleucine and leucine.

Examples of di - peptides glutamine and glutamic acid with other amino acids include the above in combination with inorganic salts of alpha-Ketoglutarate acid with di - and tripeptides.

Examples of the physical solutions of α-Ketoglutarate acid and its salts with at least one amino acid include, but are not limited to, the physical solution of at least one of the components of the group consisting of alpha-Ketoglutarate acid and its salts of sodium, potassium, calcium, magnesium with any of the following substances: glutamine, glutamic acid, arginine, ornithine, leucine, isoleucine, lysine and Proline, and any combination called the s amino acids.

The molar ratio of the alpha-Ketoglutarate acid or its salts and amino acid or amino acids of the above-mentioned physical solution should in General be in the range from 1:0.01 to 1:2, preferably from 1:0.1 to 1:1.5 and most preferably from 1:0.2 to 1:1,0.

Enter the dose varies depending on the applied active component or components of the disease, age, sex, weight, etc. of the patient, but should in General be in the range from 1 to 1000 mg/kg of body weight per day or from 10 to 400 mg/kg of body weight per day, preferably from 10 to 100 mg/kg of body weight per day.

The invention will be further illustrated by several examples, which should not be construed as limiting the scope of invention.

EXAMPLE

History:Surgical removal of the stomach (gastrectomy, Gx) leads to osteoporosis in humans and animals. Gastrectomy mainly damages the structure of spongy bone. It is unclear, does she also negative effects on epiphyseal cartilage. Dietary alpha-Ketoglutarate (AKG) is the source of hydroxyproline, the richest in amino acid in the bones and cartilage of Pro-collagen. The purpose of this study was to elucidate the effect of AKG on the bone/cartilage destruction associated with gastrectomy.

Ways:they used 40 female rats of sprage-Deuli. Twenty rats were subjected to gastrectomy and sec the Lena river into two groups: Gx+AKG and Gx+Placebo. Other twenty rats were subjected to simulated operations and is divided into two other groups: Imitation+AKG and Imitation+Placebo. After 8 weeks the animals were euthanized and were selected skull, femur and tibia. Were established bone density (BMD) and mineral saturation bone (BMC) in the right femoral bones and the bones of the leg and held histomorphometry left bones. Were also performed measurements of skulls on transilluminated images.

Results: dietary alpha-Ketoglutarate has a strong protective effect on the damaged bone of the skull of rats subjected to gastrectomy. AKG shows strong antirestriction action on the cells of the epiphyseal cartilage, the volume of cancellous bone and the shape of the trabeculae rats underwent gastrectomy.

Conclusion:AKG minimize damage to bone and cartilage associated with gastrectomy in rats.

Surgical removal of the stomach leads to osteopenia and arthritis in humans, rats and other experimental animals. People gastrectomy is associated with osteopenia. Dysfunction of the stomach may also contribute to the development of osteoporosis with age. Therefore, in most studies related to bone disease, involved patients after gastrectomy.

Gastrectomy affects mainly of spongy bone, and sometimes also of tubular bones, including what I mentioned effect on the destruction of the bones of the skull. There is information on the reduction of the mass of the tube and spongy bones in people of both sexes. The volume of the cancellous bone of the femur and tibia is reduced by 60% after 16 weeks after gastrectomy. Bone loss increases the risk of fractures of the hip, spine and other body parts of patients who underwent gastrectomy, which is a serious problem at present.

It is assumed that the loss of bone mass in patients undergoing gastrectomy, is not the result of malnutrition or lack of gastric juice or vitamin D. the Mechanism gastrectomy-induced osteopenia is still unknown. However, it is assumed that the main cause of osteoporosis is ineffective the resistance of the collagen of the bone after the mass destruction of his osteoclasts. The main component of bone Pro-collagen is a Proline - amino acid synthesized in the gastrointestinal tract of AKG and through Proline, which in turn is transformed from bone Pro-collagen in hydrocephaly in the presence of AKG, vitamin C and Fe2+. Recently it was shown that AKG effective to prevent bone loss in rats subjected oophorectomy, and in denervated bones of turkeys. Considering the above, the main purpose of this research was to establish whether food AKG to prevent loss of bone and cartilage mass in rats subjected to gastrectomy.

Animals and surgery

Forty female rats of sprage-Deuli at the age of 10 weeks (220-230 g) were kept in Macrolon cages® (2 rats in each cage) and received power from a standard dry food for rats (Lactamin, Vadstena, Sweden) and placebo or AKGad libitum,dissolved in water (table 1). The study lasted 8 weeks. Rats every week weighed.

Rats drank from 25 to 50 milliliters each day. It can be generally assumed that the rats drank from 10 to 20% of body mass.

Rats from group AKG drank approximately 25 ml AKG drink per day. In 25 ml of AKG drink contained 0.36 g AKG, which gives about 1 to 1.4 g AKG on weight kg rat per day. Rats in the placebo group (control group) drank approximately 50 ml of the drink-placebo per day.

Surgery

Twenty rats were subjected to gastrectomy and divided into 2 groups: Gx+AKG and Gx+placebo (10 rats in each group). The glandular part of stomach (i.e. the part that produce acid, fundus and pyloric cavity) were cut, after which the non-ferrous division (cardiac part of stomach) was connected to the duodenum, the end-to-end. 20 rats were subjected to a simulated operation and divided into two groups: Imitation+AKG and Imitation+Placebo. Imitation operations included abdominal incision in the midline, manipulation of the stomach and closed the ie section. Anesthesia was applied by subcutaneous injection of Ketalar® (50 mg/kg; Parke-Davis, Morris Plains, NJ, U.S.A.) and Stresnil® (40 mg/kg; Janssen-Cilag Pharma, Vienna, Austria). For analgesia was used subcutaneous injection of Temgesic® (0.18 mg/kg, Schering-Plough, Kenilworht, NJ, U.S.A.). Treatment groups Imitation+Placebo and Gx+Placebo was conducted by applying a placebo, while the group Gx+AKG and Imitation+AKG received treatment AKG.

Rats Gx every second week (starting from the first week after surgery) received intramuscular injection of 0.4 mg/kg vitamin B12 (Betolvex® 1 mg/ml, Dumex, Copenhagen, Denmark), in order to compensate the internal factors that affect the absorption of vitamin B12, and 20 mg Fe3+/kg of polymaltose complex hydroxide of iron (Ferrum 50mg Fe3+ mg/ml, Vifor (International) Inc., St. Gallen/Switzerland) as an additive, a warning poor absorption of iron due to loss of gastric juice.

These additives did not influence the development of body weight of the rats did not undergo any surgical procedures.

During the experiment 8 animals died. The number of animals (n) was 7 in the group Imitation+Placebo, 10 in group Imitation+AKG, 8 in group Gx+Placebo and 7 in group Gx+AKG.

All animals were euthanized by opening the abdominal aorta under anesthesia mentioned above.

Studies were approved by the local Committee for the protection of animal rights, Lund, Sweden.

Selection and analysis of tissues

The skull of each rat was dissected and PTS is converted from soft tissues by careful removal of the periosteum. Drying was achieved by coating each skull gauze soaked in saline solution, and placing them in an airtight container at 4°C until examination. Each skull was placed in a glass Petri dish on top of the light source (industrial lighting pipes), radiating the light of constant intensity. The resulting transilluminated images were taken using a camera attached to a surgical microscope, magnification ×16. Images were analyzed on histomorphometrical the computer made ImageJ v. 1.33a. Was estimated loss of bone mass (when the observation area of the gaps).

And femoral and tibial bone were selected and stored in 70% ethanol until further analysis.

The right femoral and tibial bones were subjected to analysis PIXIMUS, which shows BMD in g/cm and BMC in g/see

Fixed with ethanol femoral and tibial bone were calcinatory 7% nitric acid for 48 hours. Samples of the distal femur and proximal tibia (consisting of epiphysis with vosjmikilometrovom part of the metaphysis) were used in further histological process. The samples were immersed in paraffin. Longitudinally slice samples of the femoral and tibial (6 μm thick) was made automatic micro is ω Microm HM 360. It was made 20 slices each bone (with a gap of 20 μm after each 5) one individual. Sections were stained haematoxylin/eosin under standard conditions. Were removed microscopic image of each colored slice. Images for assessment of the trabecular bone were taken using a Nikon Eclipse E800 light microscope, magnification ×40 and Nikon D70 digital camera. Microscopic images of sections of the femoral and tibial bones were subjected histomorphometrical computer analysis. Analysis of trabeculae was done using ImageJ v. 1.33a. Images to assess the state of epiphyseal cartilage were made by contrasting technique Norman and selected feature AXIOVERT 200 M with LSM 5 Pascal laser scanning head, Zeiss, magnification ×100, with argon laser with a wavelength of 514 nm. Analysis of the epiphyseal cartilage was done using Analysis v 3.0. Images of the articular cartilage were obtained using fluorescence mode AXIOVERT 200 M with LSM 5 Pascal laser scanning head, Zeiss, magnification ×100, with argon laser with a wavelength of 514 nm. Images of the articular cartilage were evaluated Zeiss LSM Image Examiner v. 3.1.0.99. Parameters estimation of trabeculae under the epiphyseal cartilage were: the volume of the trabecular bone (BV/TV %), measured to characterize the trabecular bone, and the fractal dimension of trabeculae (Box Counting Method). The estimation parameters epiphyseal cartilage would the and: the number of cartilage cells in the ROI (area of interest), consisting of a zone of peace, proliferative zone and hypertrophic zone of the cartilage. Assessment of the relative content of collagen in the articular cartilage was done by measuring the fluorescent intensity of eosin-stained collagen in randomly selected ROI (with the same area of each slice with 6 plates 83 μm in diameter each throughout the articular cartilage), with the detector LSM 5 Pascal laser scanning head with a 12-bit grey scale as a scale of measurement. The dimension of each slice were conducted in exactly the same standard conditions.

Statistics

Data were verified using univariate analysis of variance (ANOVA), t-test, t-test and p<0,05 was considered statistically significant.

RESULTS

At the end of the experiment the body weight of animals subjected to surgery was 8% less than in animals subjected to simulated operations. A statistically significant difference between the groups was not (Figure 1).

Transilluminated images of the skull

Transilluminated images of the skull show a significant increase in percentage of bone lacunae in the groups of rats Gx+placebo and Gx+AKG compared with groups Imitation+Placebo and Imitation+AKG (Figure 2). In the group of rats Gx+AKG was also revealed significantly lower percentage of gaps compared with the group Gx+placebo. (*p = 0,031) (Fig.) The difference between groups Imitation+Placebo and Imitation+ AKG was not statistically significant.

Bone density (BMD) and mineral saturation bone (BMC) in the femoral and tibial bones

In the groups Gx+placebo and Gx+AKG BMD and BMC was lower than in groups Imitation+Placebo and Imitation+AKG (data not shown). However, BMD in the group Gx+AKG tended to be higher than in the group Gx+placebo (p=0,19).

Histomorphometry

Analysis of articular cartilage

The amount of cartilage collagen in the group Gx+AKG was the same as in the control (subjected to simulated operations groups, and was significantly higher compared with group Gx+placebo (table 2).

Analysis of the epiphyseal cartilage

Quantitative assessment of cell growth epiphyseal cartilage showed a decrease in the number of cells in rats Gx+AKG compared with groups of rats Imitation+Placebo and Imitation+AKG. In addition, the number of cartilage cells in the group Gx+AKG was significantly greater than in both groups of simulation (table 3, 4).

The amount of spongy bone

The volume of the trabecular bone was reduced in the groups Gx+placebo and Gx+AKG compared with groups Imitation+Placebo and Imitation+AKG. However, the reduction in the area of trabeculae in Gx+AKG was less than in Gx+placebo group (table 5, 6).

The fractal dimension of the bone trabeculae

The fractal dimension of the bone trabeculae in Gx+AKG was similar in the control groups and was higher than in Gx+placebo (table 7, 8).

EXPLANATION

The aim of the EC is periment was to evaluate the effect of dietary alpha-Ketoglutarate on bone loss, caused by gastrectomy. The obtained data confirmed this hypothesis. Really food AKG prevents the loss of bone and cartilage mass in rats subjected to gastrectomy. Our results are consistent with data from recent experiments showing that AKG prevents osteoporosis in rats subjected to oophorectomy in women after menopause.

Gastrectomy caused the loss of cartilage collagen and cartilage cells in the groups Gx+placebo, but not in the group Gx+AKG. In the group Gx+AKG was confirmed by the presence of 22% more cartilage cells than in Gx+placebo group. This suggests that AKG effective to prevent the loss of cartilage mass in rats subjected to gastrectomy. The analysis revealed a protective effect of AKG on bone and cartilage collagen. The amount of collagen in the group Gx+AKG was within range of the control group of the experiment was approximately 18% higher than the group of rats Gx+Placebo.

Observed protective effect of AKG on the bones of the skull rats underwent gastrectomy. In turtles rats group Gx+AKG showed 20% less damage than turtles in rats of group Gx+Placebo. Values of BMD and BMC showed that gastrectomy caused osteopenia in Gx+placebo and Gx+AKG groups, which is consistent with other experiments. However, using more sensitive histomorphometrical technology, we showed that KG, possibly effective for the prevention of osteopenia in Gx rats.

Further investigation of the amount of bone trabeculae showed 38% less reduction in rats Gx+AKG compared to the group of animals Gx+Placebo. Moreover, the fractal dimension of trabeculae in the group Gx+AKG showed almost the same level as in groups of simulated operations. Thus, alpha-Ketoglutarate really has a strong influence on the renewal of bone trabeculae.

Gastrectomy has a strong destructive effect on the skeleton, causing osteopenia and arthropathy. AKG cannot completely stop the damage, but it certainly limited the development of destructive changes of the bone and cartilage associated with gastrectomy, and probably contributed to the update of the skeletal system. The results of these studies may be important for clinical observations of people, for example, when a partial gastrectomy is recommended for weight reduction in patients suffering from obesity. All of these patients will develop osteoporosis and arthropathy. Thus, the introduction of these patients food AKG can stop or limit the destructive changes of the bones.

Table 1
Composition drinks AKG and placebo
IngredientsAKG
Placebo
(g/dm3)
AKG (alpha-Ketoglutarate)14,60
HCl hydrochloric acid03,32
With6H12O6glucose30,030,0
C12H22O11sucrose15,015,0
NaOH sodium hydroxide3,63,6
KOH potassium hydroxide0,750,75
Ca(OH)2calcium hydroxide0,460,46
Mg(OH)2magnesium hydroxide0,180,18
pH4,64,6

To achieve the same pH level in each solution drink placebo was titrated with 0,1M HCl to pH 4.6 ( pH AKG-drink).

Table 2
The effect of AKG and gastrectomy on the relative content of collagen in the articular cartilage
The treatmentFluorescenceSD
Gx+AKG2363a623
Gx+Placebo1928b647
Imitation+AKG2475a457
Imitation+Placebo2171a374

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

Table 3
The effect of AKG and gastrectomy on the number of chondrocytes in the epiphyseal cartilage of the hip
The treatmentThe number of cells/mm2SD
Gx+AKG2220a490
Gx+Placebo1760b360
Imitation+AKG1890b330
Imitation+Placebo1850b220

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

Table 4
The effect of AKG and gastrectomy on the number of chondrocytes in the epiphyseal cartilage of the tibia
The treatmentThe number of cells/mm2SD
Gx+AKG2470a470
Gx+Placebo1950b330
Imitation+AKG1840b410
Imitation+Placebo2110b340

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+is lacebo

Table 5
Influence of alpha-Ketoglutarate and gastrectomy on the volume of the trabecular bone of the thigh
The treatmentThe area of the trabeculae (%)SD
Gx+AKG18,8a3,7
Gx+Placebo11,2b2,1
Imitation+AKG25,5c7,8
Imitation+Placebo24,5c5,9

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

Table 6
Influence of alpha-Ketoglutarate and gastrectomy on the amount of spongy bone in the tibia
The treatmentThe area of the trabeculae (%)SD
Gx+AKG16,7a3,4
Gx+Placebo10,5b2,5
Imitation+AKG24,9c5,3
Imitation+Placebo21,1cthe 5.7

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

Table 7
Influence of alpha-Ketoglutarate and gastrectomy on the fractal dimension of the femur
The treatmentThe area of the trabeculae (%)SD
Gx+AKG1,22b0,02
Gx+Placebo1,19a0,03
Imitation+AKG1,24b0,04
Imitation+Placebo1,25b0,03

Different letters shown in the result column, anywayt significantly different at p< 0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

Table 8
Influence of alpha-Ketoglutarate and gastrectomy on the fractal dimension of the tibia
The treatmentThe area of the trabeculae (%)SD
Gx+AKG1,22b0,02
Gx+Placebo1,17a0,02
Imitation+AKG1,22b0,03
Imitation+Placebo1,21b0,04

Different letters shown in the result column, describe significantly different at p<0,05.

n=7 in Gx+AKG, n=8 in Gx+Placebo, n=10 in Imitation+AKG, n=7 in Imitation+Placebo

The explanation of the drawings:

Figure 1 Effect of dietary alpha-Ketoglutarate and gastrectomy on the body weight of rats. Control group: IMITATION+PLATZ, GX+PLATZ. Experimental group: IMITATION+AKG, GX+AKG (IMITATION rats subjected to simulated operations, GX - rats subjected to gastrectomy).

Figure 2 Selected images of skulls judge the pilot animals. Control group: IMITATION+PLATZ, GX+PLATZ. Experimental group: IMITATION+AKG, GX+AKG (IMITATION rats subjected to simulated operations, GX - rats that underwent gastrectomy).

Figure 3 Effect of dietary alpha-Ketoglutarate and gastrectomy for transilluminated images of the skull. Control group: IMITATION+PLATZ, GX+PLATZ. Experimental group: IMITATION+AKG, GX+AKG (IMITATION rats subjected to simulated operations, GX - rats that underwent gastrectomy).

1. The use of pharmaceutically acceptable salts of alpha-Ketoglutarate acid to obtain a pharmaceutical composition for the treatment or prevention of cartilage lesions in conditions including weight loss and/or impaired nutrition, malnutrition, gastrectomy, partial gastrectomy or ligation of the stomach.

2. The use according to claim 1, in which the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is a salt of an alkaline or alkaline earth metals or combinations thereof, selected from the salts of sodium, potassium, calcium, magnesium.

3. The use of claim 1, wherein the pharmaceutical composition further comprises at least one amino acid.

4. The use according to claim 3, in which the amino acid is selected from glutamine, glutamic acid, arginine, ornithine, leucine, isoleucine, Proline and combinations thereof.

5. The use according to claim 2, where the pharmaceutical is acceptable salt of alpha-Ketoglutarate acid is a salt of alkali metals.

6. The use according to claim 2, where the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is a salt of alkaline earth metal.

7. The use according to claim 1 where the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is alpha-Ketoglutarate sodium.

8. The use according to any one of claims 1 to 7, in which the input to the patient a dose of the substance is in the range from 1 to 1000 mg/kg body weight/day.

9. The use according to any one of claims 1 to 7, in which the input to the patient a dose of the substance is in the range from 10 to 400 mg/kg body weight/day.

10. The use according to any one of claims 1 to 7, in which the input to the patient a dose of the substance is in the range from 10 to 100 mg/kg body weight/day.

11. Method for the treatment or prevention of cartilage lesions in conditions including weight loss and/or impaired nutrition, malnutrition, gastrectomy, partial gastrectomy or ligation of the stomach, where the method includes the introduction of a pharmaceutical composition containing pharmaceutically acceptable salt of alpha-Ketoglutarate acid.

12. The method according to claim 11, in which the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is a salt of an alkaline or alkaline earth metals or combinations thereof, selected from the salts of sodium, potassium, calcium, magnesium.

13. The method according to claim 11, in which the pharmaceutical composition additionally contains at least the bottom of the amino acid.

14. The method according to item 13, in which the amino acid is selected from glutamine, glutamic acid, arginine, ornithine, leucine, isoleucine, Proline and combinations thereof.

15. The method according to item 12, where the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is a salt of alkali metals.

16. The method according to item 12, where the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is a salt of alkaline earth metal.

17. The method according to claim 11, where the pharmaceutically acceptable salt of alpha-Ketoglutarate acid is alpha-Ketoglutarate sodium.

18. The method according to any of § § 11-17, in which the input to the patient a dose of the substance is in the range from 1 to 1000 mg/kg body weight/day.

19. The method according to any of § § 11-17, in which the input to the patient a dose of the substance is in the range from 10 to 400 mg/kg body weight/day.

20. The method according to any of § § 11-17, in which the input to the patient a dose of the substance is in the range from 10 to 100 mg/kg body weight/day.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to pharmaceutics and medicine, and concerns application of sophoricoside.

EFFECT: preparing the drug applicable for preventing and/or treating articular cartilage degeneration or osteoarthritis in postmenopausal females, exhibiting side-effect-free high efficacy.

6 cl, 11 dwg, 5 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology, and represents a sterile injectable water-based composition in the form of gel for iintra-articular injection containing hyaluronic acid of molecular weight 0.1 to 10×106 Da in the amount of 1-100 mg/ml of water or one of its salts and optionally one or more other natural polysaccharides specified in a group consisting of chondroitin sulphate, keratan, keratan sulphate, heparin, heparan sulphate, cellulose and its derivatives, chitosan, xanthan, alginate and their salts, and also one or more polyalcohols in the amount of 0.0001-100 mg/ml of water, prepared by making a water-based formulation containing hyaluronic acid or one of its salts, optionally one or more other natural polysaccharides, and also one or more polyalcohols, sterilising it with wet steam; the prepared gel shows a coincidence frequency of the elasticity modulus G' and the viscosity modulus G" within the range 0 to 10 Hz preferentially 0.41 Hz ±0.41 Hz with G" exceeding G' if the coincidence frequency is found to be high.

EFFECT: invention provides higher degradation resistance of the gel, and also makes the gel visco-elastic.

11 cl, 8 ex, 11 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine and pharmaceutical industry, particularly to a method for making a drug for osteoarthrosis and to a method of treating osteoarthrosis. The method for making the herbal preparation for osteoarthrosis applied for the purpose of electrophoresis wherein grinded dry raw herbs, including wild rosemary shoots, elecampane inula rhizomes and roots, common St. John's wort herb, wild camomile blossom, cowberry rhizomes and roots, black poplar buds, hop cones taken in certain proportions, extracted in 70% ethanol under certain conditions; the extracts are combined, purified by double separation, boiled out in vacuum, dried up in a spray dryer; the dry powder is used to prepared 20% herbal preparation in 25% aqueous dimethylsulphoxide. The method of treating osteoarthrosis aiding electrophoresis wherein the herbal preparation described above is applied on a pad to perform an electrophoresis procedure with modulation diadynamic currents.

EFFECT: preparation made by the method described above, and a method for diadynamic-current electrophoresis are effective for osteoarthrosis.

2 cl, 1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine. Substance of the method for rehabilitation of patients suffering osteoarthrosis involves pelotherapy and herbal medicine. The method applied a mixture of dry extracts of cowberry (5 portions), lucerne (5-10 portions), dimethylsulphoxide (10-15 portions) added to peat peloid (up to 100 portions). The composition and peat peloid is applied in layers in osteoarthrosis not associated with synovitis: by applying the composition on the injured joint in a layer 0.5-1 cm coated with peat peloid to the thickness of 2-3 cm, at temperature of the composition and peat peloid - 38-42°C, duration of procedure 20-30 minutes, every second day or two days running with a pause for one day, therapeutic course - 10-15 procedures. In osteoarthrosis associated with moderate synovitis: by applying the composition on the injured joint in a layer 0.5-1 cm coated with peat peloid to the thickness of 2-3 cm, at temperature of the composition and peat peloid - 20-24°C, duration of procedure 15-20 minutes, every day or two days running with a pause for one day, therapeutic course - 8-10 procedures.

EFFECT: presented method of the combined use of herbal and pelotherapy provides higher effectiveness of rehabilitation of the patients with osteoarthrosis and simplifies the method.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, and may be applied for the purpose of stimulating cartilage hyaline neogenesis accompanying early destructive diseases. That is ensured by recovering and culturing self-specific multipotent mesenchymal stem cells (MMSCs) from fatty tissue. Then said cells are mixed with the gel carrier Lintex-Mesogel. The prepared cell suspension in gel is introduced into the joint.

EFFECT: method provides effective cartilage hyaline neogenesis without fibrocartilage formation due to optimal selection of the cell material and the specified gel carrier promoting maintained intraarticular viability, proliferative activity and MMSC chondrogenic potentialities.

2 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to azaadamantane derivatives of formula (I), to their pharmaceutically acceptable salts possessing the properties of nAChR ligands, their application, a method of treating and based pharmaceutical compositions, and also to intermediate compounds of formula (VI) and (VII) and to application of the compound of formula (V) for preparing the compound (I). In general formulas

L1 represents -O- or -NRa-; A represents -Ar1 or -Ar2-L2- Ar3; Ar1 represents 5-9-member heteroaryl wherein said heteroaryl is optionally substituted by alkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, carboxy, carboxyalkyl, cyano, halogenalkoxy, halogenalkyl, halogen, hydroxy, nitro, -NH2, (NH2)carbonyl and oxido; Ar2 represents 5-6-member heteroaryl wherein said heteroaryl is optionally substituted by alkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, carboxy, carboxy alkyl, cyano, halogenalkoxy, halogenalkyl, halogen, hydroxy, nitro, -NH2 and (NH2) carbonyl; Ar3 represents aryl, optionally substituted alkoxy, alkoxyhalogenalkyl, alkyl, aryl, halogenalkoxy, halogen, hydroxy and -NH2; or Ar3 5-9-member heteroaryl wherein said heteroaryl is optionally substituted by alkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, aryl, carboxy, carboxyalkyl, halogenalkyl, heterocyclyl and tritylaryl; L2 represents a bond, -O- or -C(O)NRa-; and Ra represents hydrogen.

EFFECT: preparing the pharmaceutically acceptable salts possessing the properties of nAChR ligands.

41 cl, 11 dwg, 162 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular orthopaedics. A whirlbone is centred; four to ten wires of the diameter 2.0 mm from a subtrochanteric region into a femoral neck. It is followed by draining irrigation of the joint with furacilin and isotonic solution. A perforator is used to form four-five canals ended in an epiphysis which are used to introduce 2 to 3 ml of the content from a medullary cavity of shin bone. A width of the joint space is adjusted by distraction.

EFFECT: local stimulation of repair processes and transosseous fixation provide restoration and reduced length of inpatient treatment.

1 ex, 8 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, particularly oncology, and may be applied in treating cancer. A method involves producing stem CD8+T-cells from a subject, introducing cladribine into the subject; contacting the CD8+ T-cells with xenogenic antigen-representing cells loaded with one or more peptide antigens with generating thereby the activated cytotoxic T-lymphocytes (CTL) which are targeted on the cells expressing said one or more peptide antigens. Then the activated CTLs and at least two cytokines which have an effect on CTL persistence are introduced into the subject.

EFFECT: use of the invention provides higher clinical effectiveness ensured by prolonged persistence of the activated CTLs and reduced number of complications, namely opportunistic infections.

10 cl, 1 tbl, 2 dwg, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new compounds of formula I or their pharmaceutically acceptable salts showing an ability to inhibit sphingosine kinase, to a based pharmaceutical composition, to a method of inhibiting sphingosine kinase and a method of treating diseases specified in breast cancer, diabetic retinopathy, arthritis and colitis. , wherein X represents -C(R3,R4)N(R5)-, -C(O)N(R4)-; R1 represents phenyl unsubstituted or substituted by 1 or 2 halogens. The values of R2, R3, R4, R5 substitutes are such as specified in the patent claim.

EFFECT: preparation of new compounds.

17 cl, 24 dwg, 9 tbl, 26 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to the use of a metal of group 4 or 5 of the periodic system specified in titanium, zirconium, hafnium, niobium and tantalum, or its oxide for preparing a drug used for treating or preventing a disease characterised by undesired expression and/or release of interferon-γ inducible protein of weight 10 kDa, IP-10 in a subject. The invention also refers to a method of treating or preventing the disease characterised by undesired expression and/or release of interferon-γ inducible protein of weight 10 kDa, IP-10, which implies the extraction of a biological fluid in a subject suffering said disease, the ex vivo contact of the biological fluid and the metal specified in titanium, zirconium, hafnium, niobium and tantalum, or its oxide, the fluid return after the contact with the metal to the subject stated above. The invention also provides the use of the method stated above or its oxide for the in vitro removal of interferon-γ inducible protein of weight 10 kDa, and a method for removing said protein. The invention also refers to a device for biological fluid purification which comprises a purification chamber with a fluid inlet and outlet and containing metal particles specified in titanium, zirconium, hafnium, niobium and tantalum, or its oxide, used for biological fluid purification, and a filter coupled with the fluid inlet and outlet to prevent said particles from escaping from the purification chamber.

EFFECT: invention provides the selective decrease of expression and release of chemokine, namely interferon-γ inducible protein of weight 10 kDa, IP-10.

25 cl, 6 tbl, 16 dwg

FIELD: medicine.

SUBSTANCE: invention refers to experimental medicine and veterinary science. The invention disclosed unexpected administration of Pro-Gly-Pro peptide as an intranasal agent in prevention and treatment of gastric ulcer in rats.

EFFECT: invention provides higher effectiveness and simplified administration of the agent.

5 ex

FIELD: medicine, veterinary science.

SUBSTANCE: invention concerns veterinary science. The invention discloses administration of Arg-Pro-Gly-Pro tetrapeptide as an agent for prevention and treatment of viral diseases. A pharmaceutical composition for prevention and treatment of viral diseases contains an aqueous solution of Arg-Pro-Gly-Pro tetrapeptide in concentration 0.1-1 % and Nipagin in a certain relation of ingredients. A method for prevention of viral diseases involves intranasal, oral or intraperitoneal introduction of said pharmaceutical composition in a dose 0.2-5 mg a day per a mouse for 2 days. A method for treatment of viral diseases involves intranasal, oral or intraperitoneal introduction of said pharmaceutical composition in a dose 0.2-5 mg a day per a mouse for 3 days.

EFFECT: invention enables extending the range of antiviral drugs, providing higher clinical and preventive effectiveness in viral infections, reducing drug cost.

4 cl, 4 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a peptide copolymer containing lysine, alanine, tyrosine and glutaminic acid residues in ratio of 0.4-0.45:0.3-0.36:0.09-0.11:0.14-0.18, including its acid addition salt - acetate, and having average molecular weight in the interval 8.7-14.5 kDa, which can be used in a pharmaceutical composition for preparing a medicinal agent against multiple sclerosis, particularly medicinal agent Glatiramer acetate. The peptide copolymer is obtained through copolymerisation of N-carboxyanhydride of Nε-trifluoroacetyl-L-lysine and/or N-carboxyanhydride of Nε-benzyloxycarbonyl-L-lysine with N-carboxyanhydrides of L-alanine, gamma-O-benzyl-L-glutaminic acid and L-tyrosine with an initiator diethylamine in a medium of aprotic solvents dioxane or tetrahydrofuran or mixtures thereof. Benzyl type protective groups are removed from lysine and glutaminic acid residues using a solution of hydrogen bromide in acetic acid with a catalytic amount of water, and volatile amines diethylamine or methylamine in form of their aqueous solution are used to remove protective trifluoroacetyl groups from lysine residues.

EFFECT: method enables control of such properties of the product as average molecular weight and amino acid composition.

3 cl, 7 ex

FIELD: medicine; veterinary science.

SUBSTANCE: administer an immunopotentiator to females of mammals into a lactemia initial stage. The immunopotentiator immunifan is administered to lactating females in a dose of mass of a body of 0.0006-0.0008 mg/kg. The immunopotentiator amyxine is administered to lactating females in a dose of mass of a body of 0.0017-0.0018 g/kg.

EFFECT: increase of content of immunoglobulins, leucocytes and mononuclear cells in milk of lactating animals.

FIELD: medicine, oncology.

SUBSTANCE: the present innovation deals with inhibiting the growth of host's cerebral tumors. It deals with introducing therapeutically efficient quantity of integrin αv antagonists being cyclo(Arg-Gly-Asp-D-Phe-[N-Me]-Val). The innovation enables to considerably inhibit cerebral oncogenesis in vivo, moreover, despite antiangiogenesis, due to induction of direct lethality of tumor cells.

EFFECT: higher efficiency of inhibition.

9 dwg, 2 tbl

The invention relates to medicine, in particular to a gastroenterologist, and can be used for the prevention and treatment of gastric ulcers in animals

The invention relates to the field of cell biology, in particular to new peptides with the ability to interact with intracellular transduction signal, thereby inhibiting the process of signal transmission, leading to proliferation and cell motility

The invention relates to compositions and methods for the reduction or regulation of inflammation and for the treatment of inflammatory diseases and other pathological disorders involving intercellular adhesion

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical and food industry, particularly producing compositions of biologically active substances applicable as biologically active additives. The pharmaceutical composition contains glycine enriched by crystal gamma-modification of glycin in amount 90-98 %, and an additive of non-toxic organic acids in amount 2-10 %. Gamma-modification of glycin makes up to 8-95 % of total amount of glycine. As additives of the organic acids, the non-toxic acids are specified as follows: malic, malonic, citric acid, citric acid hydrate or a mixture thereof. A method for preparing the pharmaceutical composition of glycine is implemented by the combined mechanical treatment of the ingredients in a vibrating mill for 6-60 minutes.

EFFECT: pharmaceutical composition of glycine under the invention is solid-stable.

2 cl, 6 dwg, 2 tbl, 9 ex

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