BG61191B1 - Pharmacologically active substance bpc, the process for its preparation and its use in the therapy - Google Patents

Abstract

The method for obtaining a human or animal BPC bodyguard gastric juice is characterized by the following stages of production: collected and homogenized animal or human gastric juice is divided into solids and liquid supernatant by centrifugation; the liquid supernatant is dialysed and lyophilized; the dry residue is purified via a chromatographic column filled with a slightly alkaline, modified ion exchange resin or gel; the concentrate is again dialysed and lyophilized; the dry residue is purified by gel- chromatography; the substance BPC is finally formed from the concentrate using dialysis and lyophilization.

Description

The invention relates to a new substance BPC, to a method for its preparation from human or animal gastric juice and to its therapeutic application.

BACKGROUND OF THE INVENTION

Gastric juice, considered as secretion of the parietal and other cells, contains a number of electrolytic components, hydrochloric acid, a number of enzymes, namely pepsin, other proteins, renin, lipase, urease and lysozyme. It contains many peptides and peptide fragments, for example the peptide hormone gastrin, and many active gastric secretion stimulants, first discovered by Adkins (I).

The natural gastric juice contains glycoproteins, which have been described (2) and intrinsic factor (IF), which is a thermally unstable mucoprotein with a molecular weight of about 60,000. This factor accelerates the absorption of vitamin B ₍₂ ).

So far no substance has been found in gastric juice with very strong anti-stress effects and other bodily protective effects.

Until now, the gastrointestinal tract, in particular the stomach, has been considered a stressed organ. The possibility of the gastrointestinal tract, preferably the stomach, being the organ that can initiate the protective response of the whole organism has never been discussed. This stress-directed response should take place in the formation and release of a new endogenous agent. Our attempts were made to isolate this agent from gastric juice. Finally, the expected substance was isolated from the gastric juice of 42 patients, which was designated the abbreviation HRV, which means a bodyguard.

SUMMARY OF THE INVENTION

The constitution of the substance BPC is very complex and, after long studies, can thus far be characterized as a folded protein with a partial sequence at the Nth end

H ₂ N-Gly-Glu-Pro-Pro-Pro-Gly-Lys-ProAla-Asp-Asp-Ala-Gly-Leu-Val-COOH

It has a molecular weight of about 40,000 ± 5,000 Daltons, determined by gel chromatography.

According to the invention, the BPC agent is prepared from human or animal gastric juice, which is initially homogenized and centrifuged, after which the supernatant is purified by dialysis, ion exchange chromatography, again by dialysis and lyophilization, and finally the BPC agent is obtained by chromatography using gel, dialysis and lyophilization.

This substance has not been described and published anywhere and has never been used as a medicine.

The study of the biological effects of BPC using many in vivo and in vitro assay techniques shows a surprisingly wide range of activities, but it is known that many diseases and pathological conditions can occur in stress or, in turn, in trauma itself or illness can cause stress. These results also suggest that BPC is produced by stress, regardless of its origin.

The BPC agent of the invention was tested in vitro u in vivo (rats, mice, rabbits and guinea pigs) and the following pharmacological properties were established:

1. Acute toxicity

Acute toxicity was determined in mice (18-25 g body weight) / sacrificed after the 30th day of the test period / of both sexes, to which HRV was administered intraperitoneally, intravenously and intragastrically. The lethal dose has not been established even with the use of such high doses as 100 mg / kg body weight, which are at least 10M ⁵ times higher than the minimum effective dose. For a long period of time, BPC was injected .2-Strengthened intraperitoneally, intravenously and intragastrically (30 mg / kg) once daily for 30 days. No pathological changes (morphological or physiological) were detected. In addition, the average effective doses are in the range of 0.01 to 50 gg / kg at parenteral administration and T1 is at least 1000.

2. Ulcers caused by immersion in water or moderate stress

Male rats (180-240 g) of the Wistar breed were used. Animals were immersed in cold water (26 ° C) for 3 hours or restricted for 48 hours. After the end of the experimental period, the stomach is examined for damage ("stress ulcers"). BPC is injected intraperitoneally and intragastrically (0.1 to 10 gg / kg) one hour or 24 hours before causing damage. On gastric lesions, HRV shows a strong protective effect.

3. Ulcers caused by the binding of the bile duct and the hepatic artery or by the introduction of cysteamine.

Ligation experiments were performed on male Wistar rats (160-250 g). The bile duct and hepatic artery are ligated. This procedure causes severe ischemic necrotic lesions of the liver tissues and ulcers of the stomach and duodenum within 6-24 hours.

Similarly, cysteamine (400 mg / kg) was administered to female rats over 24 hours to induce duodenal ulcers.

BPC is administered intraperitoneally at doses of 0.1 to 10 gg / kg. Large ulcers caused by the described literature of the stomach and duodenum are largely prevented in animals treated with HRV. Similarly, cysteamine-induced ulcers are also prevented by pre-administration of animal BPC to animals.

4. Effect of myocardial infarction induced

Male Wistar rats (180-240 g) were immobilized for 24 hours. The animals were then treated with indomethacin (25 mg / kg subcutaneously) at the start of immobilization and at 12 hours thereafter.

BPC (50 gg / kg) was administered one hour before the procedure and at the 6th and 18th hours of restriction.

In the second group, the animals were restricted for 48 hours. After these procedures, they are sacrificed and the enzyme values are determined.

The resulting protective effect of HRV is noted.

In these attempts, the protective effect is very pronounced. In contrast, no protective effects were observed for isoprenaline-induced cardiac damage. Isoprenaline (30 mg / kg, i.p.) was administered at the beginning of the trial and 24 hours later.

BPC (50 gg / kg, i.p.) was administered one hour before isoprenaline administration. Animals were sacrificed 24 hours after re-treatment with isoprenaline and 20 tested.

5. Effect on blood pressure

Male rats (180-220 g) anesthetized with urethane were used. The carotid artery is zeroed and blood pressure is recorded with a dynograph. BPC is given intravenously into the jugular vein at doses of 1 Ogg to 5 mg / kg. The results indicate no changes in the blood pressure of animals in the rat group prepared as described. Short-term changes in blood pressure are induced by the intravenous administration of noradrenaline, adrenaline, 5-HT, acetylcholine or isoprenaline. Animals pretreated with 35 BPC at doses of 10 gg to 5 mg / kg did not alter the blood pressure change caused by the vasoactive agents used.

6. Impact on damaged liver The experiment was performed on male Wistar rats (180-250 g). The lower part of the bile duct and the hepatic artery are connected. This intervention causes severe ischemic necrotic damage to the liver tissue at intervals of 6 to 24 hours. BPC is administered intraperitoneally at doses of 0.1 to 10 gg / kg. It prevents to a great extent (eg, one hour before binding) or completely eliminates damage (eg, injected at least 50 hours after ligation). In addition, the protective effect on the liver of BPC administered intraperitoneally at doses of 10 gg / kg body weight one hour before intoxication of rats with tetrachloromethane given at doses of 0.1 to 10 ml / kg intragastrally or intraperitoneally is demonstrated. BPC (0.01 to 10gg / kg, intraperitoneally or intragastrically) also prevents fatty liver changes caused by restriction stress for 48 hours.

7. Action on damaged pancreas

Male rats (170-240 g) of the Wistar breed were used. Damage to the pancreas is caused by ligation of the bile duct at the point of its infusion into the duodenum. This procedure causes clear pancreatic necrosis 24 hours later and about 50% of the rats die during this experimental period. BPC, administered at doses of 0.1 to 10gg / kg intraperitoneally, exhibits a strong dose-dependent protective effect. A significant beneficial effect on survival (82%) was demonstrated.

8. Effect on damaged kidney a / unilateral nephrectomy

The experiments were performed with albino rats (190-250 g) of Wistar breed of both sexes. Unilateral nephrectomy is performed. Weight gain of the remaining kidney was evaluated in the control and treated with BPC (10.0 µg / kg live weight intraperitoneally one hour before the procedure) groups at 24 hours after surgery. BPC has been shown to greatly reduce the increase in organ weight of the remaining kidney.

The biochemical parameters are comparable in both groups. Therefore, it seems that BPC really improves the function of the remaining kidney tissue.

b / gentamicin administration

The experiments were performed with male Wistar rats. The lesion of the canals is accomplished by the administration of gentamycin at a dose of 40 mg / kg intraperitoneally (once daily) for 30 days. All animals were sacrificed five days after the last treatment. In the control group, animals treated with gentamicin and brine showed canal damage. In the group of BPC-treated pathohistological manifestations are significantly less.

9. Effect on experimental diabetes

Male rats (175-230 g) of the Wistar breed were caused by diabetic melitus with streptozocin or oloxan. BPC at doses of 10 to 100 µg / kg, administered intraperitoneally, decreases streptozocin-induced diabetes. Moreover, HRV significantly increases the survival time of the animals.

10. Effect on temperature

The antipyretic activity of BPC was demonstrated in male Wistar rats (180-220 g) using brewery yeast injections. Pretreatment with BPC administered in single doses (5-20 gg / kg, intraperitoneally) one hour before the yeast injection significantly reduced the increase in temperature measured at 30-minute intervals for at least three hours.

11. Effect on caragenin-induced edema

Experimental swelling of mice (18-24 g) from both sexes was caused by injection of carrageenin into the hind paws. BPC is effective at doses of 10 gg / kg intraperitoneally when injected one hour before the experimental procedure, reducing the volume of edema caused by the indicated agent. In addition, BPC at the same doses was inactive in the hot plate experiment (53.5 ° C) on mice.

12. Effects on experimental rhinitis

Wistar rats are exposed to vapor of 10% formaldehyde for one hour daily for 15 days. In the experimental group, BPC was administered at doses of 10 gg / kg, intraperitoneally, every day one hour before exposure to vapor. Animals are sacrificed after 13-15 days of treatment. In BPC-treated animals, edema of the nasal mucosa was significantly smaller and the epithelium was preserved compared to the control group. The secretion is catarrhal and not purulent.

13. Effect on experimental fractures

The effect of BPC (10 gg / kg, intraperitoneally) on experimental fracture was investigated in the healing process of shin bone fractures in rats. With regard to controls, histological examination indicates a significant increase in the speed of the healing process in all rats4.

Μ sc, «..- τ- · j * W * e.» ** r · ** · · - * · ·. · '·> 1Α6ll9l hoves treated with BPC. In addition, they have significantly smaller local hematomas than controls.

14. Effect on experimental burns

The experimental method of burning skin or nasal mucosa was performed in mice (20-25 g). No purulent secretion was observed in comparison with the control group (treated with saline) and an increase in the rate of healing was observed in all BPC pre-treated (IO pg / kg, intraperitoneally, one hour before the experimental procedure) animals.

In addition, in animals with nasal injuries there are also only very moderate post-traumatic nasal bumps. Therefore, nasal respiration is only slightly impeded and survival is not impaired. These results are in clear contrast to the data obtained from the control groups.

15. Effect on skin wounds

The experimental procedure was performed with Wistar rats (170-210 g). Skin wounds are caused by wide cuts of the skin. BPC was administered intraperitoneally at single doses of IO pg / kg prior to injury. In contrast to the control groups, the treated animals showed a lack of fertilization and swelling, poor granule formation, and an increased speed of the healing process.

16. Neuropharmacological activity a / Neuropharmacological examination in accordance with the Irving methodology shows that even when HRV is administered at doses thousands of times higher than those that have proven effective (therapeutic) in many different models (20,0 and 40 (0 pg / kg body weight, intraperitoneally or orally) did not cause a significant change in posture compared to that of control mice.

b) isolated rat heart and rabbit HRV administered at 30 pg / ml doses of Ringer-Locke solution did not cause any change in the intensity and frequency of contractions (Langendorf preparation).

c) in an isolated ileum model (rat, guinea pig or rabbit) (Magnus preparation), BPC administered at the same dose had no effect on intestinal tone and on contractions caused by acetylcholine, histamine and serotonin.

17. Action of BPC on a delayed type 5 hypersensitivity reaction (DTH) to DNFB

The DTH response model for DNFB on the ear or paw of NMRI mice was used. Animals that were pre-treated with two doses of DNFB for sensitization had significantly thicker ears and paws than control animals. Mice pre-treated with BPC intraperitoneally, at a dose of 15 0.1 pg / 10 g body weight one hour before the administration of the experimental DNFB dose, showed significantly smaller differences between treated and untreated ears / paws.

18. Effect on pupil size

In the mouse experiment (18-22 g) of both sexes, BPC dripped into the left eye. The pupil size is measured using monocular lenses. In addition, a group of animals were treated with HRV introduced intraperitoneally. The results indicate that BPC locally attached does not cause myosis, and BPC injected intraperitoneally causes pupil contraction by about 20%. This effect lasts for at least 30 minutes.

19. Impact of BPC on column lesions

Mouse contact hypersensitivity is induced by administration of DNFB (2,4-dinitrofluorobenzene). Mice were given two doses to increase the sensitivity of 25 l of 0.5% DNFB (4 to 1 acetone / olive oil ratio) on the pre-shaved skin of the abdominal wall for two consecutive days. 72 hours after the last dose was administered to increase the sensitivity of the mouse, a 20 μ 20 sample of 0.2% DNFB transrectally in anesthetized ether was given. Animals were sacrificed 24 hours later and a colon (colon) was examined. Pathological changes in the colon are expressed in severe bleeding and ulceration. These lesions occupy all parts of the wall of a column in some animals, so that perforations can be noticed.

Administration of BPC (0.1 pg / 10 g) one hour after the DNFB sample significantly reduces the extent of lesions on the column. The most common lesion is petechial hemorrhage.

20. Impact on tumor cells a / Two cell lines 1-924 and melanoma B-16 were cultured in vitro under standard conditions. In PC, in vitro growth assay for said cell cultures was tested. Two days after cell culture initiation, BPC was added at 3% and 0.3% concentration (0.1 ml). Three days after this procedure, the cell count in each culture was determined using a cell counter. The results indicate that BPC has an inhibitory effect on both cell lines, eg 1.924 and B-16 melanoma.

b. The Ehrlich ascites tumor (EAT) is a tumor that develops in all breeds of mice and can grow in ascites or solid form depending on the pathway of tumor cell introduction. Studies have been conducted to determine whether incubation of EAT cells in BPC solution would alter the survival time of mice after subcutaneous or intraperitoneal injection of the EAT cells thus treated.

Animals were monitored for a 45-day period. A control group of 15 NMRI male mice was also injected with 0.4 χ 10 ⁶ EAT cells. Prior to injection, the tumor cells were incubated for one hour at 4 ° C in brine. The injection volume is 0.2 ml. The average survival of this group of animals is 36 days and only 3/15 of them survive the end of the experimental 45-day period.

An experimental group of 15 NMRI male mice received the same dose of tumor cells in the same volume, but the cells were pre-incubated in BPC solution (2 pg / 1 ml). Only 2/15 of the mice die within the observed 45-day period and the others survive more than 45 days. The difference between the control and experimental groups was significant (p <0.01).

If the same procedure was repeated and EAT cells were injected intraperitoneally (instead of subcutaneously) no difference was observed between the groups.

It can be concluded that BPC prolongs the survival of animals with tumors and that it has antitumor activity.

21. Impact on irradiation-induced damage

The experiments were performed with mice (64) of both sexes of the NMR-Y strain (22-28 g). The first control group (16 mice) was not treated and irradiated.

The second control group (16 mice) was pre-treated with saline solution (0.2 ml) and irradiated with a 9 Gy (Co-60) over-lethal dose.

The two test groups (16 + 16 mice) were irradiated with the same 9 Gy over-lethal dose. One hour after irradiation of the first group, BPC was administered at a dose of 20 pg / kg body weight. However, the second trial group was treated with the same dose of HRV but one hour before irradiation.

Post-irradiation HRV had no effect on the survival of mice, but administration after irradiation increased the survival rate after 12 days by 68.75% compared to the other experimental group.

22. Influence on the hematopoietic system

On albino mice were applied cytostatics (endoxan, Vincristine, Adriablastin, cytosine-arabinoside) in LD _J0 doses intraperitoneally. In the experimental group, BPC was administered one hour before cytostatics at a dose of 10 pg / kg, and only saline was injected into the control group. The group of animals were sacrificed on the 3rd, 5th, 7th and 11th days of the experiment. Blood tests are performed / E, Hb, Htc, L, Tb, ab. number of neutrophils /, bone marrow and cytological and histological examinations of the liver and spleen. On day 3 of the experiment, there were no differences between the test and control groups with respect to L, E, Tb and neutrophil values. But while hematopoietic cells were still intact in the bone marrow of BPC-treated animals, aplasia was already observed in the control group. The number of neutrophils and L significantly increased on the fifth day of the experiment to normalize on the seventh day of the experiment. In the control group, normalization was observed near day 11.

23. HRV and fertility - influence on oligoastenospermia

Observations were made on 10 men, aged 30 to 40 years, diagnosed with oligoastenospermia. These patients collect semen after 3 to 4 days of abstinence. After liquefaction (30 min), a layer of 0.5 ml of medium was added to 0.5 ml of semen. The control medium consisted of NAP F10 with 10% deactivated chordal serum. The test medium contains an additional 2 gg / ml or 4 gg / ml BPC. After a period of 90 min at 37 ° C in an atmosphere containing 5% carbon dioxide, the Horwell Fertility chamber determined the number of progressively motile, in-situ motile and immobile sperm in 1 ml of semen. Preliminary results indicate that at lower concentrations, BPC has no effect on sperm motility. At a concentration of 4 gg BPC / ml, a significantly higher percentage of motility was found compared to the control group.

24. Action of BPC on reproduction processes

The effects of BPC on reproduction have been studied in mice with a prior history of three pregnancies. 20 days after interruption of lactation, the animals are caged. BPC is given at a dose of 10 gg / kg, intraperitoneally, once daily for the entire duration of pregnancy (19-21 days) and lactation (for the next three weeks). The control group received an equal volume of brine at the same time. The number of offspring of each female and the weight of newborns is recorded daily.

Statistical analysis confirms a consistent significant increase in the number of newborns from each female and a surprising lack of differences in their weights throughout the observation period.

All female animals were allowed to recover for 25 days after stopping lactation. Then they are again pooled to study the effect on the fifth pregnancy.

In all animals treated with BPC compared to the control group, there was an increase in fertility and lactation capacity.

No pathologic changes in fertility rate were observed in the offspring (regardless of the number of maternal pregnancies).

Therefore, it is likely that BPC can have better fertility rates, even in fairly adult mice. Obviously, the results obtained are consistent with the overall protective action of BPC and with the in vitro assay data obtained using human sperm.

25. Protective effect of HRV in periodontal tissue injury.

The purpose of this study is to investigate the effect of BPC on the root resorption of periodontal soft tissue caused by mechanical injury of the first molar in albino rats.

Thirty rats at 4 weeks of age were allocated to a control group receiving saline and to an experimental group treated with BPC. The following procedures are applied to all animals - a needle (0.6 mm thick and 1.9 mm long) is inserted as a tool to inflict mechanical injury, which is inserted at a depth of 1 mm for 1 s from the mesial corolla margin in almost parallel to the mesial surface of the first maxillary right molar crown.

After mechanical injury, the test group was treated with BPC (0.02 ml / 200 g). Rats were divided into 12 groups and sacrificed at intervals of 1, 3, 5, 6, 14 and 21 days after the last treatment.

The right maxilla of all control and experimental animals was removed and fixed in 10% neutral buffered formalin, demineralized with formic acid, then paraffin-coated and separated into 6 gm mesiodistal sections. The sections were stained with hematoxylin and eosin and examined with a light microscope.

The histological findings in the first molar of rats in the control group are as follows - a large number of inflamed cells is detected. A small resorption cavity is observed between 1-3 days in the part of the root surface at the level of the alveolar edge. On the fifth day, the resorption cavity expands in the direction of the crown and contains numerous odontoclasts. On the seventh day, the resorption cavity with many odontoclasts expands further to the crown. On the 14th and 21st days, the cavity is further expanded to the crown, bringing the part of the cavity closer to the cement-enamel junction.

The test rats had the same changes between days 1 and 5 as controls, but they did not have progression of root resorption between days 14 and 21. In conclusion, BPC has a protective effect in the mechanical injury of the rat molar periodontal tissue.

26. Impact of HRV on major cell and spinal cord lesions

Acquired dementia of the Alzheimer's type (DAT) is characterized by an acquired complete disorder of higher functions of the cerebral cortex, which affects memory and cognition. The post-mortem study of patients with DAT shows that major cellular / NB / neurons have undergone deep selective degeneration. These neurons are the main source of external cholinergic stimulation of the cerebral cortex. The purpose of the study was to investigate the effect of BPC on the behavior of rats with bilateral electrolyte damage to NB. In particular, rats with NB lesions may serve as an experimental model for DAT.

The study was performed with albino rats. All groups (with or without NB lesions) were tested for passive abstinence using the Ashford and Jones method. Animals with affected NB received saline (control group) or BPC 10 pg / kg solution, intraperitoneally (test group).

The test substances are injected: a) only once, immediately after damage to the NB or b) once daily for four consecutive days after the damage to the NB has been done. HRV is also administered once daily for four test days, one hour before the start of the test. After the 20-day postoperative recovery period has passed, each animal is tested for its behavior. Statistical significance was calculated in agreement with the analysis of variants combined with the Duncan test for multiple comparisons (p <0.05). The survey results show the following:

- bilateral electrolyte damage disrupts passive abstinence in rats;

-BRS at the doses used does not affect passive abstinence in uninjured, unaffected animals;

-BRS can improve behavior in NB-damaged rats, significantly.

27. Influence of BPC on the motor activity of rabbits with spinal cord injuries

The purpose of this study is to investigate whether HRV can alter the effects of a spinal cord injury. In this experiment, adult rabbits of both sexes were used. Under the influence of phenobarbital anesthesia, an intermediate spinal C-L laminectomy is performed. A measurable compact spinal cord injury is applied using the technique of Albin and the team. The motor activity of the hind limbs is monitored according to the Tarlov system once daily for nine days. The main observations are as follows: 1 / complete paraplegia; 2 / minimal movements; 3 / the animal is able to stand but cannot run; 4 / the animal can run with some elasticity and uncoordination and 5 / normal movement activity.

Animals received saline in control and BPC at doses of 10 Hg / kg intravenously, once daily for nine days after surgery.

Spinal cord injury causes almost complete paraplegia. The mean values of complete (nine days post-trauma) motor activity studied in animals treated with saline alone were 1.3 units of Tarlov (N = 4) and in animals treated with BPC 2.5 units in Tarlov / N = 7 /.

It is also apparent that BPC improves the motor activity of rabbits with spinal cord injury.

From the results obtained from this experience, it can be concluded that BPC can be a promising drug in the treatment of impaired nerve function (for brain or spinal cord injury) and that BPC should be given to patients as early as possible after neuronal damage.

28. Influence of BPC on bowel motility, baseline temperature and diuresis

Administration of BPC has no effect on diuresis, decreases baseline temperature in vivo, and ability to contract smooth muscle of the intestine at. * * «** '» »-4t rV. - Mr.

guinea pig in vitro experiments.

29. Activity against Trichinella spiralis

Administration of BPC concentrate to experimental mice (DI) at doses of 10 pg / kg (body weight, intraperitoneally) pre-infected with an oral dose of Trichinella spiralis larvae has a significant impact on animal survival.

30. Breeding animals for commercial purposes

1419 healthy pigs (out of 4306 pigs) received a BPC of 10 pg / kg body weight, intramuscularly, immediately after birth, another 1440 on the 13th day after birth, one day before castration, and the remaining 1477 received only the usual iron-containing therapy. Injuries, scrapes, mortality, body weight, food intake are assessed after 4 weeks. HRV applied only once significantly reduced the rate of rejection (in the HRV treatment group immediately after birth) and the rate of injury (in both HRV treated groups). Groups of animals treated with BPC achieved the same body weight as controls but with lower food consumption. Infection effects - 40 weaned pigs infected with E. coli-induced enterocolitis at 28 days of age were examined. BPC was administered to 20 animals (10 pg / kg body weight, intramuscularly). After one month, there was a significant decrease in mortality in the HRV-treated animals.

In another experiment, mortality rates after the first week of life in a total of 1,200 healthy pigs were examined. 400 animals received HRV immediately after birth in addition to conventional Fe-therapy. A significant decrease in mortality was observed in BPC (10 pg / kg body weight, intramuscular) treated animals.

31. Antivirus activity

Antiviral activity was tested in vitro and in vivo (mice). BPC is used at a dose of 10pg / kg body weight, intraperitoneally or T.e.V. Significant activity was observed against enteroviruses (Eco type-6,9,11 and 16; Coxsackie type-A6, B3 and B4), CNS viruses (Ixod encephalitis, LCM choriomeningitis) and ARBO viruses (Tahyna, Bhanja and Calovo). Survival prolongation (72 hours for absolute survivors) was noted in inf10 mice.

Some aspects of the invention

Technical problem. The technical problem is whether or not it is possible to affect pharmacotherapy at the same time.

I. 1. Acute hepatitis (viral, drug-induced, toxic, acute, regardless of etiology); 2. Chronic hepatitis; 3. Liver cirrhosis; 4. Cholestasis; 5. Cholelellithiasis; 6. Cholangitis; 7. Liver transplants - treatment before and after transplantation; 8. Treatment before and after surgery, in other liver operations (eg congenital malformations, benign or malignant bile duct stenosis);

II. 9. Prevention of inflammation of the pancreas; 10. Improvement in the treatment of the inflammatory process of the pancreas; 11. Decrease in the incidence of postoperative pancreatitis; 12. Prevention of complications and mortality from acute pancreatitis; 13. Reduction of the frequency of chronic pancreatitis;

III. 14. Acute haemorrhage in the upper gastrointestinal tract; 15. Erosive gastritis; 16. Acute ulcer of the stomach and duodenum / '' stress ulcer ''; 17. Chronic peptic ulcers; 18. Non-peptic ulcers of the gastrointestinal tract;

IV. 19. Shock (regardless of its etiology);

V. 20. Prevention of renal parenchyma from the development of acute and chronic necrosis due to ischemia and hemorrhage; 21. Acute and chronic renal failure and any cases that may cause them;

VI. 22. Skeletal injuries / fractures /;

23. Skeletal healing operations (osteotomy);

24. Skeletal grafts; 25. Pseudoarthrosis; 26. Arthrodesis; 27. Wounds / increases healing speed /; 28. Burns (of the skin and mucous membranes, accelerates the healing process);

VII. 29. Anti-tumor activity;

VIII. 30. Anti-inflammatory, anti-edema, anti-rheumatic activity; 31. Antipyretic activity; 32. Rheumatic arthritis; 33. Osteoarthrosis; 34. Ankylosing spondylitis; 35. Urical arthritis; 36. Extra-articular rheumatism (bursitis, tendonitis, synovitis, humeroscapular periarthritis); 37. Inflammation and edema after surgical and non-surgical treatment, fractures and excisions;

IX. 38. Acute myocardial infarction; 39. Myocarditis; 40. Elimination of ischemic changes in tissues;

X. 41. Reduced food consumption;

XI. 42. Hyperprolactinemia;

XII. 43. Parkinson's disease, parkinsonism;

XIII. 44. Glaucoma (reduction of intraocular pressure-activity similar to that of dopamine and pilocarpine);

XIV. 45. Diabetes mellitus;

XV. 46. Barbiturate intoxication;

XVI. 47. Stress and all adverse events that develop as a consequence of stress.

State of the art

There is still no such universal and specific pharmacotherapy and the treatment of complications is beyond it.

Solution to a technical problem

The digestive tract, in particular the stomach, has so far been considered only as an organ for the effects of stress. The possibility of suggesting that the gastrointestinal tract, in particular the stomach, may be an organ from which a response from protective bodies may be elicited has not been discussed. A substance that triggers, propagates and controls the body's deterrent response, later called BPC, is isolated from the gastric juice of 542 patients and from animal sources by dialysis, ion exchange filtration and gel chromatography. Therefore, BPC (although its structure has not yet been fully elucidated) has been tested in various models of human disease, with in vivo and in vitro experiments. On the basis of the results obtained, it can be expected to be successfully administered over an extremely wide range in pharmacotherapy.

The substance, isolated and called BPC, is unknown and has therefore not been used in pharmacotherapy.

The resulting BPC was studied in various experimental species (eg rats, mice, guinea pigs) in vivo, as well as in human cells in vitro.

BPC is called so based on the abbreviation for the term "Body Protecting Compound".

Causing damage to the liver, pancreas, stomach and duodenum.

Medium laparotomy is performed under the influence of mild ether anesthesia. The bile duct then connects peripherally to the duodenum at the point where the hepatic artery (with the same surgery simultaneously connects to the bile duct) reaches it on the left side, just below the portal vein, which remains intact in said ligature. The wound is then closed and the animal remains without being disturbed until its natural death or until its death.

Under the influence of mild ether anesthesia, an average laparotomy is performed and the bile duct connects just before it enters the duodenum. The wound is then closed and the animals are left undisturbed until their natural death or death.

Animals were immersed in water at a temperature of 26 ° C with a depth of 20 cm for 3 hours. Immediately afterwards, they are killed. 24 hours before being immersed in water, the animals are left without food with free access to water.

Under the influence of mild ether anesthesia, an average laparotomy is performed and the pylorus is connected. The wound was closed and the animals were left untouched until their death, only fifteen minutes after the ligation of the pill.

Animals are immobilized for 24 hours. Indomethacin was administered concomitantly with immobilization and 12 hours later. After 24 hours, the animals were killed.

Causing fractures, wounds, skin burns and mucous membranes

Under the influence of mild ether anesthesia, a fracture of the tibia is caused by digital pressure using a hypomochion. Animals were sacrificed on the 5th, 8th, 9th, 12th and 30th days after fracture.

Widespread anesthesia causes a wide wound to the skin. Animals are sacrificed daily during the first five-day period, then three days in the period up to the fifteenth day, and then five days after the monthly period of injury.

Under the influence of mild anesthesia with ether, a burn wound is made on the back of the live tube (15 χ 30 mm) with a glowing heat-generator for 15 seconds. Animals are sacrificed on the fifth, ninth and thirtieth day after the lesion is caused.

Under the influence of a mild ether anesthesia, the nasal mucosa is burned bilaterally with a glowing heat sink for five seconds. Animals are sacrificed on the fifth, ninth and thirtieth day after the injury is caused.

Causing kidney damage

Under the influence of mild anesthetic anesthesia, tying of the kidney is carried out. 24 hours later, the animals are killed. After the wound is closed, some of the animals are left undisturbed until their natural death.

Causing anti-inflammatory, anti-inflammatory and antipyretic effects

A solution (1%) of carrageenin (0.05 ml subcutaneously) was applied to the paw as an inflammatory agent; paw volume was measured three hours later according to the usual procedure.

A synoenzyme (2.5 g / kg, i.p) was used to assess body temperature. The rectal temperature is then measured for three hours.

Causing heart damage

Animals are immobilized for 24 hours. Indomethacin was administered at the onset of immobilization (0 hours) and 12 hours later. After 24 hours, the animals are killed.

Isoprenaline (30.0 mg / kg, i.p) was administered at the zero hour of the experiment and at the 24th hour. At the end of the 48-hour period, the animals are killed.

Causing CNS effects

-the influence on the spontaneous behavior of animals (mice) is studied.

To determine the presence of dopamine neurons in the activity of BPC and CNS, the usual dopamine agonist apomorphine (20 mg / kg, subcutaneously) as well as sirotonin (5 mg / kg, intraperitoneally) (comparative controls) or saline (0, 9% sodium chloride) (control). After the administration of these substances, the behavior of the animals is studied. Stereotypic behavior is assessed by the method of Malik (1983).

Studying the effects on the iris muscle

The test solution is administered either locally or intraperitoneally. Measure the pupil diameter at appropriate intervals (5, 10, 15, 30, 60, 90 and 120 minutes) using the usual procedures.

Studying the effect on the depressive activity of barbiturates

Anesthesia was caused by administration of sodium pentobarbital (50 mg / kg intravenously). The duration of sleep caused by the barbiturator is estimated by the presence or absence of the so-called "equilibrium reflex".

Causing effect on diabetes mellitus

Diabetes mellitus was caused by the administration of alloxan (100 mg / kg intraperitoneally) as well as streptozotocin (100 mg / kg, intraperitoneally). The glucose content of urine and blood is measured by conventional methods. Animals are left untouched until their natural death.

Impact assessment

Distribution

Studies with HRV, labeled 1-125, show an accumulation of HRV, which is also dose- and time-dependent, exactly in the stomach in most cases, spreading throughout the body after oral or parenteral use.

Clinical observations

In studies dealing with damage to the liver, pancreas, stomach and duodenum, kidney, heart, fractures, wounds, burns (skin and mucous membranes), diabetes mellitus, the overall condition of animals treated with HRV is significantly better with that of control animals. In contrast to the control groups, there was either no mortality in the BPC-treated groups or a decreased mortality rate was firmly observed.

Macroscopic and pathological observations

In control animals with bile duct ligature and hepatic artery, extensive yellow-green necrosis of the liver, sometimes including all parts of the liver, as well as pancreatic necrosis and lesions of the haemorrhage of the stomach and duodenum develops. The application of

BPC successfully and depending on the dose prevents the described lesions. In control animals, these lesions occur approximately 6 hours after the ligation, whereas in BPC treated animals, they are not visible even after 48 hours after the ligation. The pathohistological examination maintains the observed changes and differences in the macroscopic examination.

In control animals with bile duct ligature, severe pancreatic necrosis and lesions of the stomach and duodenal hemorrhages occur at the point of entry into the duodenum. The administration of dose-dependent HRV prevents the development of the lesions described. The pathohistological examination maintains the macroscopic observed changes and differences.

In control animals exposed to the stress of immersion in water, the stress of immobilization and administration of indomethacin, as well as short-term pyloric ligation (15 minutes) showed significant gastric lesions. The introduction of HRV significantly reduces (or completely inhibits) the development of gastric lesions following the adverse procedures described above.

In control animals with a renal ligature (canal), serious renal lesions were detected. A significant reduction in lesions was observed in HRV-treated rats. Pathohistological studies firmly confirm macroscopically observed changes and differences.

Compared to control animals with scrotal fracture in BPC-treated animals, the development of post-traumatic hemorrhage was significantly less pronounced; animals used a functionally injured leg earlier. Qualitative pathohistological studies using conventional procedures show an increased and accelerated rate of bone healing in BPC-treated rats throughout the observation period.

In control animals with wounds, extensive development of purulent granulation was observed during the first days after injury. In BPC-treated rats, unlike controls, the wound is dry, with no secretion and visible granulation. Pathohistological analyzes maintain macroscopically observed changes and differences throughout the observation period.

In control animals subjected to restrictive stress and indomethacin, myocardial damage with elevated serum CPK was observed. The introduction of HRV significantly reduces myocardial damage and decreases IBS values. Pathohistological analyzes confirm macroscopic observations of differences and changes.

In animals with experimentally induced diabetic melitus, BPC abolishes transient glycosuria in alloxan-treated groups. In animals treated with streptozotocin, administration of HRV almost threefold increased survival. In all animals treated with HRV, polydipsis and polyphagia were less pronounced.

Anti-inflammatory, anti-edema and antipyretic effect

There was a marked decrease in carrageenin-induced edema in animals treated with HRV compared to controls during all observation periods.

In animals treated with sickoenzyme, the introduction of HRV significantly reduced the increase in body temperature compared to the results observed in the control group.

Effects

Results of in vitro analysis

Two days after the start of cultivation of human fibroblast L-929 and melanoma B16 cells, BPC was added. Three days after that, the cell count was calculated with a cell counter. BPC shows a dose-dependent toxic effect on melanoma cells. No effect on fibroblasts.

When comparing the control animals with skin burns with the BPC treated animals, a significantly faster crust development was observed in the treated animals. In control animals with burns of the mucosa (eg, the mucous membrane of the nose), severe swelling of the damaged tissues was observed during the first two days, followed by purulent secretion and partial obstruction of breathing through the nose. Development of the described changes was not observed in the animals treated with HRV. Pathohistological analyzes confirm12 await macroscopic observed differences and changes.

Effects on CNS

The effect of BPC on the CNS is examined by observing animal behavior (mice)

Five minutes after administration, an increase in animal motility was observed. Their anxiety increases from minute to minute and peaks in 30 minutes. At that moment, the animals stand on their hind legs, erect like a candle, and choke. The strobesphenomenon is obvious, the animals are running around the cell, and the piloorection is obvious. After apomorphine administration, the change occurs after 45 minutes and is accompanied by partial standing of the hind legs. Following serotonin administration, only sterotypic behavior (increased movement, shortness of breath, less intense washing) is noted for a short period.

Topical or intraperitoneal administration results in myosis (20% approximately).

The administration of HRV shortens the barbiturate sleep, which is established by shortening the time without a “rebalance reflex” (by at least 60%).

Influence on blood pressure, bowel motility, baseline temperature and diuresis

Administration of BPC has no effect on blood pressure changes caused by adrenaline, noradrenaline, isoprenaline, acetylcholine and serotonin, and on normal blood pressure values.

The administration of HRV does not affect the smooth muscle of the intestine (guinea pig).

The use of BPC does not reduce the normal temperature.

The use of HRV does not increase diuresis.

Mechanism of action

It may be suggested that the action of BPC, like other exogenous proteins, is due to one of the mechanisms — the direct interaction of the protein or its fragments with the cellular receptors, stimulating or inhibiting the release of biologically active (stimulating or inhibiting) proteins or non-proteins by re10 receptor or non-receptor related mechanisms, interaction with enzymes that break down endogenous biologically active peptides. The numerous protective effects on otherwise unrelated nosological objects could be explained if they refer to the general term stress (unfavorable) that causes them in the body. It seems likely that BPC is the first deterrent (starting from the stomach) against any stressful event that threatens the body. Therefore, the mechanism of action of BPC is the primary defense mechanism against various adverse attacks against the body.

From the foregoing, it can be concluded that it is possible to exert a pharmacologically beneficial effect on the development of the following disorders by administering BPC: I. 1. Acute hepatitis (caused by viruses, medicated, toxic, acute, regardless of its etiology); 2. Chronic hepatitis; 3. Liver cirrhosis; 4. Cholestasis; 5. Choledocholithiasis; 6. Cholangitis; 7. Liver transplantation - treatment before and after transplantation; 8. Preoperative and postoperative therapy for other liver operations (eg congenital malformations, benign and malignant cholecoccus stenosis)

II. 9. Prevention of pancreatitis; 10. Improving the development of pancreatitis; 11. Decrease in the degree of postoperative pancreatitis; 12. Prevention of complications and mortality in acute pancreatitis; 13. Reduction of cases of chronic pancreatitis;

III. 14. Acute bleeding at the top of the digestive tract; 15. Erosive gastritis; 16. Acute ulcers, stomach and duodenum (stress ulcers); 17. Chronic peptic ulcers; 18. Non-peptic ulcers of the gastrointestinal tract;

IV. 19. Shocks (with different etiology);

V. 20. Protection of renal parenchyma against necrosis caused by ischemia or hemorrhage (acute or chronic); 21. Acute and chronic renal failure, and all causes thereof;

VI. 22. Skeletal injuries (fractures); 23. Treatment procedures on the skeletal system (osteotomy); 24. Skeletal grafts; 25. Pseudoarthrosis; 26. Arthrodesis; 27. Wounds (faster healing); 28. Burns (on skin, mucous membranes, faster healing);

VII. 29. Antitumor effect

VIII. 30. Anti-inflammatory, anti-edema and anti-rheumatic effect; 31. Antipyretic effect; 32. Rheumatic arthritis;

33. Arthrosis; 34. Ankylosing spondylitis; 35. Gout; 36. Extra-articular rheumatic disorders (bursitis, tendonitis, synovitis, humeroscapular periarthritis); 37. Inflammation and edema after surgical and neurosurgical procedures accompanying fractures and dislocations;

IX. 38. Acute myocardial infarction; 39. Myocarditis; 40. Delay of ischemic tissue changes;

X. 41. Reduction of food consumption (anorexitive effect);

XI. 42. Hyperprolactinemia; 43. Parkinson's disease;

XII. 44. Glaucoma (reduction of intraocular pressure-effect of dopamine and pilocarpine);

XIII. 45. Diabetes mellitus;

XIV. 46. Barbiturate intoxication;

XV. 47. Stress and all stress-induced damage to the body.

The administration of BPC can be either oral or parenteral, in the form of pills, capsules, dragees, tongue tablets, suppositories, vaginal preparations; injection solutions, nasal sprays; ointments, creams, gels, sprays, eye drops, ear drops and nose drops.

Embodiments of the invention Preparation examples

Example 1. Method for the isolation of BPC

The material from which BPC was isolated was obtained from the gastric juice of 542 patients repeatedly and reproductively and from animal sources.

It is stored at 2 to 8 ° C and homogenized. It was then centrifuged (12000 G / 30 min) and the supernatant collected. 200 ml of it was dialyzed against water (at + 4 ° C) at a ratio of 1 to 125 until a conductivity of 100 pS was reached. The resulting dialysate was lyophilized. The dry residue was dissolved in 10 ml of double-distilled water and chromatographed on a column filled with modified DEAE ion exchange gel (40 to 80 µm). The column measures 25 x 250 mm. A buffer solution (sodium acetate 0.01 M-ene, pH 5.2) was used for dilution. The fraction was detected using a UV (UV) detector within 2800 nm. 40120 ml fractions were collected. The resulting eluate was first concentrated using AQUACIDE-3 (Calbiochem, USA) and then lyophilized.

The dry matter was dissolved in 10 ml of 0.2 M pH 7.4 phosphate buffer solution and then fed to a SPHACRYL-S-200 chromatographic column, 2.5 x 450 mm in size, at a speed of at a flow rate of 1.2 ml / min. 40 to 120 ml fractions were collected.

The solution was dialyzed against distilled water again, concentrated with AQUACIDE3 and lyophilized. Approximately 200 mg of dry white solid is obtained.

Characterization

BPC is characterized by ultraviolet (UV) and infrared (IR) spectra (Figures 1 and 2).

Based on the pharmacological properties listed, it can be used in the treatment of certain diseases, especially when the natural formation of this agent in the body is insufficient or even lacking.

It can be used to treat a) all stress-related illnesses and disorders;

b / circulatory disorders - ulcers (of different origin), shocks (of different etiology), ischemia and heart attacks (of different organs);

c / inflammation and edema of any etiology, hepatitis (acute and chronic), cirrhosis of the liver, cholangitis, pancreatitis (with different etiology), gastroduodenitis, nephritis and nephropathy, myocarditis, pericarditis, endocarditis, arthrosis and arthrosis (arthrosis) , rhinitis, rheumatic disorders - disorders of connection with different etiology, temperature, lesions caused by free radicals;

d / traumas and surgical interventions traumatism (pre- and mid-traumatic treatment), grafts, wounds, burns, fractures, post-surgical complications;

e / malignancies - some types of tumors;

e / diabetic melitus;

g / disorders with dopaminergic or similar etiology-fullness, Parkinson's disease, hyperprolactinemia, glaucoma, sleep caused by barbiturates (barbiturate poisoning);

h / Diseases and disorders of the nervous system - Alzheimer's disease, ischemic and traumatic damage to the nervous system and delay in mental degenerative changes;

and / in dentistry - diseases and disorders of the oral mucosa, surgical treatment of periodontal pockets, inhibition of root resorption;

j / hematopoietic system - neutropenia, aplastic anemia, myelodysplastic syndrome in bone marrow transplantation;

k / radiation damage caused by acute radiation syndrome, protection against radiation damage;

l / immunology - hypersensitivity reactions, allergies and transplants;

m / fertility - increase in fertility, treatment of oligoastenospermia and induction of lactation;

n / in veterinary medicine for the industrial rearing of animals for the reduction of mortality, for crippling and injury, for improving food absorption and weight gain;

o / in Virology - antiviral therapy;

n / miscellaneous - fatty changes in the liver, cholechtases, cholelithiasis, acute and chronic renal failure, toxic damage to the kidney and liver.

BPC can be used as a medicine for enteral, topical, rectal or parenteral use. BPC-based pharmaceutical preparations are prepared by the addition of inorganic or organic excipients. For the preparation of tablets and dragees, for example talc, lactose, starch, stearates and the like are added, and for solutions and suspensions, for example, water, alcohols, glycerines and the like are added. For suppositories, for example, natural and hardened oils, waxes or polyethylene glycols are added. Pharmaceutical formulations may contain suitable preservatives, buffers, stabilizers, surfactants, solubilizers, sweeteners and various coloring agents.

Suitable daily doses for BPC are from 5 to 50 pg / kg body weight and can be administered in a single dose or in divided doses.

Example 1. Preparation of HRC from human gastric juice

The crude material was obtained reproducibly from the gastric juice of 542 patients. The cooled material should be stored at 2 to 8 ° C and homogenized before use.

Adequate portion of this material (200 ml) was first separated by centrifugation (12000 G for 30 min) into solids and a liquid supernatant. This fluid was dialyzed against demineralized water (1 to 125) at 4 ° C until a conductivity of 100 pS was reached. The solution was then lyophilized. The dry residue was dissolved in 10 ml of distilled water and fed to a chromatographic column (25 x 250 mm) filled with DEAE-modified ion exchange gel (eg Trisacryl M-DEAE, ^R IBF). Sodium acetate buffer (0.01 M, pH 5.2) was used for elution. The fractions were monitored with a 280 nm ultraviolet detector. Fractions of 40 to 120 ml of the eluate were collected, concentrated using AQUACID3 ^R (Calbiochem) and then dialyzed and lyophilized.

The dry substance was dissolved in 10 ml of 0.2 M-phosphate buffer (pH 7.4) and purified again by feeding into a SephacrylS-200 ^R gel (Pharmacia Fine Chemicals) in a chromatographic column (2.5 x 450 mm). ). The flow rate was (1.2 ml / min). The collected fractions from 80 to 200 ml were re-dialyzed against distilled water, concentrated using AQUACID-3 and then lyophilized. This gives 2 mg of a white substance. The resulting BPC is very soluble in water. It decomposes at temperatures in the range of 180 to 200 ° C. Its molecular weight, determined by gel chromatography, is 40,000 ± 5,000 D. The ultraviolet spectrum of BPC (in water) shows absorption in the regions shown in Fig. 1.

The infrared spectrum of BPC (in potassium bromide) shows absorption bands as shown in Figure 2.

BPC has a folded protein structure with a partial amino acid sequence from the N-terminus:

HjN-Gly-GIu-Pro-Pro-Pro-Gly-Lys-ProAla-Asp-Asp-Ala-Gly-Leu-Val ..... COOH

Example 2. Preparation of HRC from human gastric juice

Homogenized human gastric juice from the same source as in Example 1 was centrifuged, dialyzed and lyophilized according to the procedures described in Example 1.

The dry substance (80.4 mg) was dissolved in 0.1 M acetate buffer (3 ml, pH 5.22) and fed to a chromatographic column (10 x 50 ml) filled with FractogelTSK ^R ion exchange resin (Merck) equilibrated with acetate buffer (pH 5.2). The flow rate is 0.7 ml / min. The fractions were monitored using a UV spectrometer at a wavelength of 280 nm. The first fraction with strong UV absorption was collected, dialyzed against distilled water and then lyophilized (47 mg).

The resulting dry matter was redissolved in acetate buffer (1 ml) and fed to a chromatographic column (16 x 560 mm) filled with Sephadex-G-100 ^R gel (Pharmacia Fine Chemicals), pre-equilibrated with acetate buffer. The flow rate was 0.3 ml / min. The collected fractions from 66 to 108 ml were re-dialyzed against water (distilled) and lyophilized. A white powder is obtained which corresponds to the product obtained and described in the preceding example.

Example 3. Preparation of the substance BPC from porcine gastric juice

Homogenized porcine gastric juice (85 ml) was centrifuged, dialyzed and lyophilized according to the procedures described in the previous example to give 440 mg of dry residue. Then, 110 mg of this dry residue was dissolved in 0.1 M acetate buffer (6 ml, pH 5.22), fed to a chromatographic column (10 x 50 mm) filled with Fractogel-TSK ^R ion exchange resin, and treated as described in Example 2. 44 mg of a yellow concentrate are obtained.

Part of this concentrate (24 mg) was dissolved in 1 ml of the same acetate buffer and fed to a chromatographic column (16 x 565 mm) filled with Sephadex ^R -50/80 gel equilibrated with acetate buffer. The flow rate is 1.35 ml / min. The eluate between 65 and 108 ml was collected, re-dialyzed against distilled water and lyophilized. The resulting white powder (10 mg) within the accuracy and reproducibility of the analytical methods used has the same molecular weight and other properties as the product described in the foregoing examples.

Example 4. Tablets

Composition mg / tablet the substance BPC 0.3 lactose 37,7 starch 6.0 talc 3.0 tragacanth 2.5 magnesium stearate 0.5 50,0 mg

Example 5. Capsules

Composition mg / capsule the substance BPC 0.3 lactose 99.0 magnesium stearate 0.7 100,0 mg

Example 6. Lingvalets (Tablets)

Composition mg / lingual the substance BPC 0.3 mannitol 100,0 starch 5.0 polyvinylpyrrolidone 1.5 talc 3.2 110,0 mg

Example 7. Suppositories

Composition mg / suppository the substance BPC 0.5 polyethylene glycol 300 478,0 polyethylene glycol 1500 529,0 polyethylene glycol 6000 38,0 emulsifier 14.5 1060,0

Example 8. Cream

Composition g / 250 g crack the substance BPC 0.05 emulsifier 25,0 hydrogenated peanut butter 50.0 Twin 60 120,0 propylene glycol 30.0 methylhydroxybenzoate 0.16 propylhydroxybenzoate 0.17 250,0 g

Example 9. Solution for injection

Composition mg / ml the substance BPC 0.3 sodium carboxymethyl- cellulose 1.5 polyvinyl pyrrolidone 5.8 lecithin 2.7 benzyl alcohol 0.01 buffer q.s. double distilled water to 1 ml

Example 10. Solution

Composition g / 25 ml substance BPC glycerin benzyl alcohol buffer double distilled water to 0.05 15.0 0.01 q.s. 25,0 ml

Example 11. Dry injection

Composition weight / mg the substance BPC 0.3 mannitol 10,0 10.3 mg

Example 12. Nose Spray

Composition mg / each dose the substance BPC 0.5 benzyl alcohol 0.15 Uliglyol ^R -812 14,0 freon 12/114 60.0 74,65 mg

Example 13. Eye drops, ears, nose

Composition mg / 5 ml the substance BPC 45 stabilizer 5 sodium hydrophosphate 116 sodium dihydrophosphate 33 benzalkonium chloride 0.5 double distilled water to 5 ml

Example 14. Forms for vaginal administration (vaginals)

Composition mg / vaginal the substance BPC 0.3 polyethylene glycol 300 150,0 polyethylene glycol 1500 1290,0 polyethylene glycol 6000 700,0 propylene glycol 200,0 Twin-40 59,7 lactose 100,0 2500 mg

Example 15. Ophthalmic ointment

Composition mg / 3.5 g the substance BPC 45 paraffin oil 1000 emulsifier 100 white petrolatum 2355 3500 mg

Claims (6)

/ Received by the International Bureau on 12 August 1991 / 12.08.91/; the original claims 1, 4 and 5 have been corrected; other claims are unchanged / CLAIMS 1. A method for the preparation of a bodyguard substance (BPC) from human or animal gastric juice, characterized by the following steps of production: a) the collected and homogenized animal or human gastric juice is separated into solids and liquid supernatant by centrifugation; b) the liquid supernatant is dialyzed and lyophilized; c) the dry residue is purified through a chromatographic column filled with a slightly alkaline and modified ion exchange resin or gel; d) the concentrate is re-dialyzed and lyophilized; e) the dry residue is purified by gel chromatography and e) the substance BPC is finally obtained from the concentrate using dialysis and lyophilization. A method according to claim 1, characterized in that in step c / a modified ion exchange gel or resin with DEAE, DMAE, QAE or with another basic group is used. A method according to claims 1 or 2, characterized in that in step e) an inorganic or organic gel with dextran, polystyrene, polyacrylamide, agarose, polyvinyl or other matrix having a molecular weight resolution of 20000 is used up to 100000 D. 4. The BPC substance obtained according to the method according to claims 1, 2 or 3, with a molecular weight of 40000 ± 5000 D, determined by gel chromatography, ultraviolet and infrared spectra and a partial N-terminating amino acid sequence of H ₂ NGly-Glu-Pro -Pro-Pro-Gly-Lys-Pro-Ala-Asp-AspAla-Gly-Leu-Val .....- COOH. Use of the BPC substance, characterized by the process for its preparation according to claims 1, 2 or 3, with a molecular weight of 40000 ± 5000 D, determined by gel chromatography, with ultraviolet and infrared spectra according to FIGS. 1 and FIG. 2 and a partial N-terminating acid sequence H ₂ N-Gly-Glu-Pro-Pro-Pro-Gly-Lys-ProAla-Asp-Asp-Ala-Gly-Leu-Val ....-COOH obtain according to claims 1, 2 and 3 for the preparation of medicaments for: a) all caused by stress, diseases and disorders; b / circulatory disorders (ulcers) of different origin, shock (with different etiology), ischemia and heart attacks (of different organs); c / inflammation and edema (inflammation and edema of any etiology), hepatitis (acute and chronic), cirrhosis of the liver, cholangitis, pancreatitis (with different etiology), gastroduodenitis, nephritis and nephropathy, myocarditis, pericarditis, endocarditis, endocarditis, endocarditis arthritis (with different etiology), rhinitis, rheumatic disorders-disorders with different etiology, temperature, lesions caused by free radicals; d / traumas and operations - traumatism (pre- and post-traumatic treatment), grafts, wounds, burns, fractures, postoperative complications; e / formations - some types of tumors; e / diabetic melitus; g / disorders with dopaminergic or similar etiology - fullness, Parkinson's disease, hyperprolactinemia, glaucoma and sleep-induced sleep (barbiturate poisoning); h / Diseases and disorders of the nervous system - Alzheimer's disease, ischemic or traumatic damage to the nervous system and delay in mental degenerative changes; and / in dentistry - in diseases and disorders of the oral mucosa, surgical treatment of periodontal pockets, inhibition of root resorption; j / hematopoietic system - neutropenia, aplastic anemia, myelodysplastic syndrome in bone marrow transplantation; k / radiation damage caused by acute radiation syndrome and prophylaxis against radiation damage; l / immunology - hypersensitivity reactions, allergies and transplants; m / fertility - increase of fertility, treatment of oligoastenospermia and increase of lactation; n / in veterinary medicine - for the breeding of animals for commercial purposes to reduce mortality, crippling and contusions, to improve food absorption and weight gain; o / antiviral treatment; n / miscellaneous - fatty liver, cholestasis, cholelithiasis, acute and chronic renal failure, toxic damage to the kidney and liver. Medicament for the treatment of diseases and disorders according to claim 5, characterized in that it comprises an active amount of BPC in pharmaceutical form for enteral, oral, parenteral, rectal and topical use. Attachment: 2 figures Literature 1. Edkins, I.S., Proc.Roy. Soc. L. 76B, 376 (1905). 2. Hanrowitz, F., Chem. and Biology of Proteins, 1950, p.199. 3. Castle et al. Am. I. Med. Sci. 178, 748 (1929).