JP2591875B2 - Pharmacologically active substance BPC, method for its preparation and method of using it for therapy - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel substance, BPC, and a method for producing it from human or animal gastric juice.

2. Prior Art Gastric juice, which is considered to be the secretion fluid of parietal cells and other cells, contains many electrolyte components, hydrochloric acid, many enzymes such as pepsin, other proteins, rennin, lipase, urease and lysozyme. . There are many peptides and peptide fragments, such as gastrin, a peptide hormone, highly active gastric secretion stimulant, etc.
Edkins: First discovered by "Proc. Roy. Soc. L.", 76B , 376 (1905).

In normal gastric juice, F. Hanrowitz: “Chem. And Biolo
Gly of Proteins ", (1950), p. 199, contains glycoproteins (mucins) and a heat-labile mucoprotein, an intrinsic factor (IF) having a molecular weight of about 60,000. It is to promote the absorption of vitamin B ₁₂ [Castle
Etc .: "Am. J. Med. Sci. 178 , 748 (1929)].

To date, no substances with very strong anti-stress action and other personal protective activities have been found in gastric juice.

Description of the Invention Heretofore, the gastrointestinal tract, especially the stomach, was only considered as a target organ for stress. The possibility that the gastrointestinal tract, preferably the stomach, may be the organ that can initiate a whole body defense response has not been discussed. This response, ie the response that is actually directed to stress, should be realized in the formation and release of new endogenous substances. We have attempted to separate this substance from gastric juice.

Eventually, the present inventors separated this expected substance from the gastric juice of 542 patients. This substance, named BPC, is an abbreviation that stands for Body Protection Compound.

The structure of this material is very complex and can be characterized by our studies to date as a folded protein with the following partial amino acid sequence from the N-terminus: H ₂ N -Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val- ‥‥‥ -COOH This is about 40,000 ± 5,000 as measured by gel filtration. Having a molecular weight of

In the present patent application, this substance BPC is made from human or animal gastric juice. The gastric fluid is first homogenized, centrifuged, the supernatant purified by dialysis, ion exchange chromatography, redialysis and lyophilization, and finally using gel chromatography, dialysis and lyophilization. This substance BPC was obtained.

So far this compound has been described everywhere,
Or it was never announced. Furthermore, it has never been used as a medicine. Studies on the biological effects of this substance BPC by various methods in vivo and in vitro have shown a surprisingly broad range of activity.

A surprisingly wide range of activity However, it is known that many diseases and various pathological conditions can be induced by stress, or alternatively that damage or disease can itself induce stress. These results also suggest that BPC is produced by stress, regardless of its origin.

The substance BPC according to the invention has been studied in the laboratory and in vivo (rats, mice and guinea pigs), whereby the following pharmacological properties were found: 1. Acute toxicity Acute toxicity was observed in male and female mice BPC was measured intraperitoneally (ip), intravascularly (iv) and intragastric (i.g., 18-25 g) (lethal after 30 days of experimental period).
g.). No lethal dose could be obtained with doses as high as 100 mg / kg body weight, at least 10 ³ to 10 ⁵ times higher than the lowest effective dose. BPC once a day for 30 days in long-term processing
Administered continuously by ip, iv and ig (30 mg / k
g). No pathological signs (morphological or physiological) were recorded. In addition, depending on the administration by ip, the average effective dose is between 0.01 and 50 μg / kg and the TI is at least 1,000.

2. Ulcer induced by water immersion or restraint stress Male rats of the Wister strain (180-240 g) were used. Place each animal in cold water (26 ° C) for 3 hours or 48
Time restrained. After the end of the experimental period, the stomach was examined for the presence of a disorder (stress ulcer). Intraperitoneal injection and intragastric administration (0.1-10 μg / kg) 1 hour or 24 hours before BPC damage induction. The strong protective effect of BPC has been shown for gastric damage.

3. Ulceration induced by ligation of the bile duct and hepatic arteries or administration of cysteamine Ligation experiments were performed on male rats of the Wister strain (160-250 g). Ligation of the biliary and hepatic arteries was performed.
This treatment, in addition to severe ischemic necrotic damage of liver tissue,
Within 6-24 hours, gastric and duodenal ulcers resulted.

Similarly, duodenal ulcers were induced during the 24 hours when cysteamine (400 mg / kg sc) was applied to female rats.

BPC was administered ip at a dose of 0.1-10 μg / kg.
The large gastrointestinal ulcer created by the above ligation
It was strongly prevented in animals treated with BPC. Similarly, cystamine-induced ulcers were also eliminated by pretreatment with BPC.

4. Effect on infarcts created on the myocardium Wister male rats (180-230 g) were fixed and immobilized for 24 hours. The animals were then treated with indomethacin (25 mg / kg, s.c.) in the same way as for immobilization and 12 hours later. One hour before this process,
BPC was applied 6 hours and 18 hours after detention (50
μg / kg).

In the second group, animals were restrained for 48 hours. After this treatment, each animal was sacrificed and various enzyme levels were measured.

The protective effect of the obtained BPC was recorded. The protective effect was very significant. In contrast, isoprenaline-induced heart damage had no protective effect. Isoprenaline was applied (30 mg / kg, ip) at 0 and 24 hours after the experiment, and BPC was administered 1 hour before isoprenaline application. The animals were sacrificed 4 hours after the second isoprenaline application and examined.

5. Effect on arterial pressure Male rats (180-220 g) anesthetized with urethane were used. The carotid artery was punctured and blood pressure was recorded on a dynograph. BPC was administered into the carotid artery by intravascular administration in an amount of 10 μg to 5 mg / kg body weight. The results showed that there was no change in animal arterial pressure in the group of rats treated as described above. Noroadrenaline, adrenaline, 5-
Intravascular administration of HT, acetylcholine or isoprenaline resulted in short lasting changes in arterial pressure. Animals pre-treated by administering BPC in an amount of 10 μg to 5 mg / kg body weight by intravascular administration did not moderate the changes in arterial pressure induced by the vasoactive substances used.

6. Effect on injured liver Experiments were performed on male Wister rats (160-250 g). Ligation of the biliary system only and the hepatic artery was performed. This treatment caused significant necrosis in liver tissue within a time period of 6 to 24 hours. 0.1 to 10 BPC
It was administered intraperitoneally in an amount of μg / kg. This strongly prevented the injury (eg, given one hour before ligation) or completely eliminated the injury (eg, injected immediately after ligation). In addition, 1 hour prior to poisoning with carbon tetrachloride administered ig or ip in an amount of 0.1-10 ml / kg body weight
Strong hepatoprotective action was shown by administering BPC ip at 0 μg. BPC (0.01-10 μg / kg, ip or i.
g.) also prevented fatty liver essence induced by stress in 24-hour restraint.

7. Effect on injured pancreas Male rats of the Wister strain (170-240 g) were used. Pancreatic damage was induced by ligating the bile duct system at the entry to the entrance to the duodenum. This treatment resulted in significant pancreatic necrosis after 24 hours and approximately 50
% Died within this experimental period. 0.1 by intraperitoneal administration
Administration of BPC at a dose of な い し 10 μg / kg resulted in a strong dose-dependent protective effect. A significant beneficial effect on survival (about 82%) was shown.

8. Effect on damaged kidney a) Unilateral nephrectomy Male and female Wistar marginal rats (weighing 190-250 g) were tested. One kidney was removed. For the control group and the BPC-administered group (10.0 μg / kg, by ip 1 hour before the treatment), the remaining kidney weight gain was evaluated 24 hours after the operation. BPC has been shown to reduce the increase in residual kidney organ weight.

Biochemical parameters were comparable in both groups. Thus, BPC appears to improve the function of the remaining kidney tissue.

b) Gentamicin administration Experiments were performed on male Wistar rats. Gentamicin in a dose of 40 mg / kg ip for 30 days (1 day
Times) created capillary damage. Last action 5
After day all animals were sacrificed. Tubule damage was found in a control group of animals treated with gentamicin and saline. In the group treated with BPC, the history expression was significantly smaller.

9. Effect on experimental diabetes mellitus Diabetes was induced with streptososin or alloxan in male rats of the Wister strain (175-230 g). i.
Administration of 10-100 μg / kg BPC at p. tended to delay streptozacin-induced diabetes. In addition, BPC significantly increased the animal's survival time.

10. Effect on fever The antipyretic activity of BPC was examined in male Wister rats (180-220 g) using brewer's yeast injection. 1 hour before yeast injection. One administration (5-20 μg / k
g, ip) pretreatment with BPC significantly reduced the temperature rise measured at 30 minute intervals for at least 3 hours.

11. Effects on Carrageenan-Induced Edema Experimental male and female mice were induced to develop experimental edema by injecting carrageenan into their hind limbs. It was effective when injected at a dose of 10 μg / kg ip one hour before the experimental treatment and reduced the volume of edema induced by the drug. In addition, the same amount of BPC differs from the opioid agonist in the hot plate test (53.5
° C).

12. Effects on experimental rhinitis Male and female Wistar rats are treated for 1 hour daily for 15 days
Exposure to 10% formaldehyde vapor. In the test group, BPC was administered intraperitoneally daily at 1 μg / k 1 hour before each exposure.
g dose. Animals were sacrificed 13-15 days after treatment. The edema of the nasal mucosa was significantly smaller in BPC-treated animals and the epithelium was retained compared to the control group. Secretion was catarrhal and not purulent.

13. Effects on experimental fractures The effects of BPC on experimental fractures (10 μg / kg, ip) were investigated in the course of treatment of tibial fractures in rats. For the animals in the control group, the historical examination showed a markedly increased rate of healing in all rats treated immediately with BPC. In addition, significantly lower local hematomas were recorded in these animals than in the control group.

14. Effect on Experimental Burns Mice (20-25 g) were treated experimentally for skin or nasal mucosal burns. No purulent patches were observed compared to the control group (treated with saline) and all
BPC treated animals (10 μg / kg, 1 hour before experimental treatment,
A consistent increase in healing rate was observed in ip).

In addition, there was very little post-lesion swelling of the nasal tip in animals with nasal mucosal injury. Thereafter, normal nasal respiration was only slightly inhibited, and survival was not inhibited. These results are evident in contrast to the data obtained for the control group.

15. Effect on skin wounds Experimental treatment was performed on rats of the Wister strain (170-210 g). A skin wound was created by making a large incision in the skin. Prior to incision, 10μ per dose
BPC was administered ip in an amount of g / kg. Slight granulation and enhanced healing rates were consistently found in the treated group due to the absence of suppuration and edema, unlike the control group.

16. Neuropharmacological activity a) From those (20.0 and 40.0 μg / kg, intraperitoneal or oral) that have been shown to be therapeutically effective in many different models by neuropharmacological studies according to Irwing BPC also showed no significant change in behavior when applied at doses several thousand times higher than in the control group of mice.

b) BPC applied in an amount of 30μ / ml Ringer's lock in isolated hearts of rats and rabbits did not cause any changes in contraction intensity and cycle (Langen
dorf specimen).

c) BPC applied in the same amount in a model of isolated ileum (rat, guinea pig or rabbit) (specimens according to Magnus) has no effect on intestinal tone and on contractions induced by acetylcholine, cystamine and serotonin. Also did not show.

17. Effect of BPC on delayed-type hypersensitivity to DNFB DT against DNFB in ears or limbs of mice of NMRY strain
A model of the H reaction was used. Animals pre-treated with the two sensitizing doses of DNFB have significantly thicker ears / limbs than animals in the control group. One hour before application of the test dose of DNFB, mice pre-treated with BPC by ip in an amount of 0.1 μg / body weight showed only a slight difference between treated and untreated ears / limbs. Did not have.

18. Effect on pupil size In experiments using male and female mice (18-22g), B
PC was instilled into the left eye. Pupil dimensions were measured with a single ocular. In addition, one group of animals was treated by intraperitoneal administration of BPC. As a result, the locally infused BP
It is shown that C causes miosis. BPC injected intraperitoneally also resulted in approximately 20% pupillary constriction. This effect lasted for at least 30 minutes.

19. Effect of BPC on intestinal disorders Contact hypersensitivity in mice was induced by the application of DNFB (2,4-dinitrofluorobenzene). To each mouse, 0.5% concentration of DNFB (in acetone: olive oil = 4: 1) was applied on the pre-shaved skin of the abdominal wall in two sensitized doses of 25 μl for two consecutive days. Mice are anaesthetized and given 0.2% concentration D rectally 72 hours after the last application
20 μl of NFB was administered. Twenty-four hours later, each animal was sacrificed and its gut was examined. Intestinal pathological changes were characterized by severe bleeding and edema formation. These injuries occupy the entire area of the intestinal wall in some animals, and can also result in perforations.

Application of BPC performed 1 hour after the test administration of DNFB (0.1μ
g / 10g) significantly reduces the appearance of intestinal damage. The most common injury was purpura.

20. Effects on tumor cells a) Two cell lines, L-924 and melanoma B-16, were cultured in vitro under standard conditions. In vitro tests were performed on the activity of BPC on the growth of the above cell cultures. Two days after the start of cell culture, BPC was added at a concentration of 0.3% and 3% (0.1 ml). After 3 days, the number of cells per culture was determined with a cell counter. The results showed that BPC had an inhibitory effect on both cell lines, L-924 and melanoma B-16.

b) Ehrlich ascites tumor (EAT) is a tumor that can grow in all strains of mice. It could grow in ascites or in solid form. This depends on how the tumor cells are administered. We attempted to determine whether culturing ETA cells in a BPC solution alters the survival time of mice after subcutaneous or intraperitoneal injection of EAT cells so treated.

We observed each animal for 45 days. A control group of 15 NMIR male mice was injected with 0.4 × 10 ⁶ EAT cells. Prior to injection, tumor cells were incubated in saline for 4 hours.
Incubated for 1 hour at ° C. 0.2ml injection volume
Met. The average survival time of the animals in this group was 36 days, and only 3 out of 15 animals survived 45 days.

An experimental group of 15 NMRI male mice received the same amount of the same tumor cells in the same application, but these tumor cells were pre-incubated in a BPC solution (2 μg / 1 ml). During the observation period (45 days) only two of the 15 mice died. The difference between the control and experimental groups was significant (p <0.01).

No difference was observed between the two groups when EAT cells were injected intraperitoneally instead of subcutaneously with the same treatment. Thus, BPC prolongs the survival of tumor-bearing animals, and it can be concluded that it has an antitumor effect.

21. Effect on radiation-induced damage Experiments were performed on male and female mice (22-28 g) of the NMR-Y strain. The first control group (16) was untreated and unirradiated. The second control group (16 animals) was a group pre-treated (0.2 ml) with saline, which was irradiated with 9 Gy (Co-60) at a lethal dose.

Both experimental groups (16/16) were irradiated at the same 9 Gy hyperlethal dose. One hour after irradiating the first group, BPC 20
It was administered in an amount of μg / kg. However, the second group was treated with the same BPC one hour before irradiation.

Administration of BPC after irradiation has no effect on the survival of mice. Administration of BPC before radiation increases survival by 68.75% after 12 days compared to other test groups.

22. dosages in cell equilibrium histologically of LD ₅₀ by intraperitoneal administration for effects on the hematopoietic system Rats (Endoxan, vincristine, Adria neublastin, cytosine - arabinoside) was applied. One hour before cell equilibrium administration in the test group, BPC
Was applied at a dose of 10 μg / kg, and saline alone was injected in the control group. Each group of animals was tested on the third, fifth,
The animals were sacrificed on days 7 and 11. Blood (E, Hb, Htc, L, T
b, absolute number of neutrophils), bone marrow, and liver and spleen anatomy and medical history were examined. On the third day of the experiment, there is no difference in L, E, Tb and neutrophil values compared to the control group. However, although hematopoietic cells were still present in the bone marrow of animals treated with BPC, aplasia was still present in animals of the control group. On the fifth day of the experiment, neutrophils and L
It rises significantly until standardized on day. No standardization occurs in the control group before day 11.

23. BPC and fertility-effects on azoospermia Ten men aged 30 to 40 years diagnosed with azoospermia were tested. Ejaculates were collected from these patients after 3-4 days of abstinence. After thawing (30 minutes), add 0.5 ml medium layer to 0.5 ml ejaculate. The control medium was HAM containing 10% inactivated cordal serum.
-F10. The experimental group medium additionally contained 2 μg / ml or 4 μg / ml BPC. Horwell Fer
9 in an atmosphere containing 5% CO ₂ in the tility Chamber
After 0 minutes of migration time, continuously mobile, in-situ mobile and non-mobile mobiles were counted in 1 ml of ejaculate at 37 ° C. Preliminary results indicate that low concentrations of BPC have no effect on sperm motility. A significantly higher percentage of motility was found at a concentration of 4 μg / ml BPC compared to the control group.

24. Effect of BPC on Reproductive Process The effect of BPC on reproduction was examined in mice with three previous reproductive histories. The animals were mated 20 days after the last lactation was completed. BPC was applied by intraperitoneal injection once daily over the entire period of pregnancy (19-21 days) and lactation (next 3 weeks) in an amount of 10 μg / kg. At the same time, the control group was given the same volume of saline. The number of offspring per female and the offspring daily were recorded. Careful statistical analysis consistently showed a significantly increased number of progeny per female and the surprisingly small differences between their body weights during each study period.

All females were allowed to recover for the next 25 days following the end of lactation. They were then mated again to determine their effect on the fifth pregnancy.

For the control group, an increase in fertility was recorded with an increase in lactation capacity in all BPC-treated animals. It should be pointed out that pathological changes, as well as changes in fertility, were not observed in BPC-treated offspring (regardless of maternal pregnancy frequency). Thus, BPC appears to be able to improve reproductive rates even in sufficiently aged mice. In conclusion, it is clear that the results obtained are consistent with the overall protective effect of BPC as well as the in vitro data obtained with human serum.

25. Protective effect of BPC against periodontal tissue damage The purpose of this study was to prevent periodontal soft tissue root resorption caused by first molar mechanical damage in white rats
Investigate the effect of BPC.

Thirty rats, 4 weeks of age, were divided into a control group treated with saline and an experimental group treated with BPC. We performed the following procedure on all animals: We used a needle-like device (width 0.
6 mm, 1.9 mm in length) and inserted for 1 second at a depth of 1 mm from around the median gingiva in a direction almost parallel to the median tooth surface of the crown of the upper right first molar.

The experimental group was treated with BPC solution (0.02 ml / 200 g) after mechanical injury. Each rat divided into 12 groups was sacrificed on day 1, 3, 5, 6, 14, or 21 days after the final treatment.

The upper right jaw of all control and experimental rats was removed, fixed in 10% neutral buffered formalin, demineralized with formic acid, embedded in paraffin and 6 μm thick in the inner plane. In addition, it cut one by one in order.

Each cut was stained with hematoxylin and eosin and examined by light microscopy. The historical findings in the first molars of rats from the control group were as follows:
The presence of numerous inflammatory cells was recorded. At the level of the alveolar crest, small resorption pits were observed in the portion of the root surface over a period of one to three days. On day 5, the resorption fossa extended in a coronal direction. This resorption pit contained a number of tooth crusts. On the seventh day, the resorption fossa containing this many tooth crust extended further coronally. On days 14 and 21, the resorption fossa extended further coronally to a portion close to the enamel boundary of the tooth.

All rats in the experimental group showed similar changes as animals in the control group for 1-5 days, but no progression of root resorption between 14-21 days. To conclude, BP
C can be said to have a protective effect on the mechanical damage of the periodontal tissue of rat molars.

26. Effects of BPC on basal ganglia and spinal cord injury Alzheimer's type dementia (DAT) is characterized by a global inhibition of higher cortical functions affecting memory and cognition. Cadaver studies in patients with DAT have shown that basal ganglia (NB) neurons have undergone significant selective alteration. These nerve cells are the main source of exogenous cholinergic innervation to the cerebral cortex. The purpose of this study was to investigate the effect of BPC on passive avoidance behavior in rats with bilateral electrolytic damage of NB. That is, rats with NB damage are DAT
Would be representative of the experimental model.

This study was performed on white rats. Ashf for all groups of rats (with and without NB damage)
A passive avoidance test was performed by the method of ord and Jones. NB-damaged saline solution was administered (control group), or 10 μg / kg of BPC solution was administered ip (test group). The test substances were injected a) only once immediately after the formation of the NB lesion or b) once a day for 4 days after the formation of the NB lesion. BPC was also administered once daily during the four days of training, one hour before the start of this passive avoidance experiment. The behavioral tests described below were performed on each animal after a 20-day recovery period following surgery. Statistical polymerizability values were calculated according to a modified method of analysis combined with Duncan's test for multiple comparisons (p <0.05). The results of the test were as follows: bilateral electrolytic damage impairs passive avoidance in rats.

-BPC at the dose used has no effect on passive avoidance behavior in undamaged intact animals.

-BPC can significantly improve passive avoidance behavior in rats with NB injury.

27. B for motor activity of rabbits with contusion in the spinal cord
Effect of PC The purpose of this study was to determine whether BPC could alter the consequences of spinal cord contusion. Male and female grown rabbits were used in this study. Dorsal L ₁ -L ₆ laminectomy was performed under pentobarbital anesthesia. A measurable compact lesion of the spinal cord was made by using the technique of Albin et al. Motor activity in the hind limbs was controlled once daily for 9 days according to the Tarlov system. Basically, classification criteria are 1) complete versus anesthesia, 2)
Minimal voluntary movements, 3) animals could get up but could not run, 4) animals could run with slight convulsions and exhibit impaired coordination, and 5) normal motor activity. .

Each animal received saline and 10
BPC at μg / kg iv was administered once daily for 9 days after surgery.

The formed spinal cord injury resulted in almost complete anti-anaesthesia. The average tested total locomotor activity (in 9 days after lesion formation) in animals receiving saline was 1.3 Tarl
ov unit (N = 4). In animals receiving BPC this was 2.5 Tarlov units (N = 7).

It is also clear that BPC improved the motor activity of rabbits with spinal cord injury.

From the results shown above, BPC may be a potential drug in the treatment of damaged nerve function (brain or spinal cord injury), and BPC should be administered as soon as possible after nerve injury It can be estimated.

28. Effects of BPC on intestinal motility, basal body temperature and diuresis The application of BPC has no effect on diuresis, reduction of basal body temperature and the ability of guinea pigs to absorb intestinal smooth muscle in vitro.

29. Activity against Trichinella spiralis 10μg / kg BPC concentrate in experimental mice (D1) which had been infected orally with Trichinella spiralis larvae in advance
Application in an amount of (ip) has a significant effect on animal survival.

30. Commercial Breeding Immediately after birth to 1419 healthy pigs (out of a total of 4306),
And one day before the castration on the 13th day
BPC (10 μg / kg, im) was administered respectively, while the remaining 1
477 animals received only normal Fe-treatment. Injuries, culling, mortality, body weight and food conversion were assessed after 4 weeks. BPC applied only once is the culling rate (BPC immediately after birth
) And in the bruise rate (in both groups treated with BPC). The same weight was obtained in animals that were BPC treated and kept at significantly lower food consumption.

Effect on infection: 40 weaned pigs, 28 days old, infected with E. coil enterocolitis were examined. BPC was applied to 20 animals (10 μg / kg, im). One month later, significantly reduced mortality was noted in BPC-treated animals.

In another experiment, mortality in the first week after birth was determined for a total of 1200 healthy pigs. 400 soon after birth
Head animals were dosed with BPC in addition to the usual Fe-therapeutic agents. B
A significantly reduced mortality was observed in animals treated with PC (10 μg / kg, im).

31. Antiviral activity Antiviral activity was examined in vitro and in vivo (mouse). By ip or Tev at a dose of 10 μg / kg
BPC was used. entero-virus (Echo type: 6, 9, 11 and 16, Coxsackie type: A9, B3 and B4), CNS virus (Ixo
Significant activity against de encephalities, LCM Choriomeningitis and ARBO viruses (Tahyna, Bhanja and Calovo) was found. Prolonged survival (72 hours to complete survival) was observed in infected mice.

In the following, some aspects of the invention are further described.

Technical issues A technical issue is whether it is possible to be affected simultaneously by drug treatment.

I 1. Acute hepatitis (irrespective of etiology, viral, drug-induced, toxic, fulminant) 2. Chronic hepatitis 3. Cirrhosis 4. Cholestasis 5. Cholelithiasis 6. Cholangitis 7. Liver transplantation-before and after transplantation Treatment 8. Pre- and post-operative treatment, including other hepatic surgical procedures (eg, congenital malformations, benign and malignant bile duct stenosis) II 9. Prevent pancreatitis 10. Improve the course of pancreatitis 11. Postoperative pancreatitis 12. Prevention of complications and death from acute pancreatitis 13. Decrease in frequency of chronic pancreatitis III 14. Acute bleeding in the upper gastrointestinal tract 15. Hot gastritis 16. Acute ulcers in both stomach and duodenum ( 17. Chronic peptic ulcer 18. Non-peptic ulcer of the gastrointestinal tract IV 19. Shock (regardless of etiology) V 20. Protection of renal parenchyma from the development of acute and chronic ischemic and hemorrhagic necrosis 21 Acute and chronic kidney damage and all the factors that cause them VI 22. Bone damage (fractures) 23. Bone treatment Placement (osteotomy) 24. Bone connection (calems) 25. False joint 26. Joint fixation 27. Wound (increased treatment speed) 28. Burn (skin and mucous membrane, increased treatment speed) VII 29. Antitumor activity VIII 30. Anti-inflammatory, anti-edema, anti-rheumatic activity 31. Antipyretic activity 32. Rheumatoid arthritis 33. Osteoarthritis 34. Ankylosing spondylolysis 35. Uric acid arthritis 36. Extra-articular rheumatism (synovitis, tendinitis, Synovitis, humeral scapular periarthritis) 37. Inflammation and edema after surgical and non-surgical treatments, fractures and dislocations IX 38. Acute myocardial infarction 39. Myocarditis 40. Delayed ischemic changes in tissue X 41. Low food consumption XI 42. Hyperprolactinemia XII 43. Parkinson's intractable disease, Parkinsonism XIII 44. Glaucoma (reduced intraocular pressure-dopamine-like and hirocarpine-like activities) XIV 45. Diabetes XV 46. Barbiturate poisoning XVI 47. All adverse effects of stress and the consequences of stress Currently there is no such general and specific pharmacotherapy yet, and the treatment of complications is out of pharmacotherapy.

Description of the solution to the technical problem The gastrointestinal tract and especially the stomach are currently only regarded as target organs of stress. The possible indication that the gastrointestinal tract, and especially gastric juice, may also be an organ in which a host defense response is initiated has not been considered. The substances that initiate, mediate, and control the corresponding biological response are termed BPCs by the inventors, which are from the gastric juice of 542 patients by the inventors, as well as from various animal sources, dialysis, Separated by ion exchange filtration and gel chromatography. Therefore this substance
BPC, although its structure has not been fully elucidated, is a variety of different models of human disease. Based on the results obtained, a very wide range of applications can be expected in drug therapy successfully.

This substance, isolated by the present inventors and termed BPC, has heretofore not been known and thereby has not previously been used in drug therapy.

We have studied the resulting substance BPC in vivo on a number of different experimental animals (eg rats, mice, guinea pigs) and in vitro on human cells.

The substance BPC is an abbreviation for "Body Protecting Compound".

Induction of liver, pancreas, stomach and duodenal damage Central laparotomy was performed under light ether anesthesia. The hepatic artery of the biliary duct (ligated simultaneously with the biliary duct by the same surgical procedure) was then ligated just below the portal vein at a distance from the duodenum where it reached from the left side (portal The pulse remains undamaged above the ligature). The wounds were then conjugated and the animals were released free until death or death.

A central laparotomy was performed under light Teether anesthesia, and the bile duct system was ligated shortly before its entry into the duodenum. The wounds were then conjugated and the animals were released free until death or death.

The animals were immersed in water at 26 ° C. for 3 hours to a depth of 20 cm. Shortly thereafter they were killed. Water immersion
Twenty-four hours before, each animal was allowed free access to water without food.

A central laparotomy was performed under light Teether anesthesia and the pylorus was ligated. The wound was sutured and each animal was released freely until sacrificed only 15 minutes after pyloric ligation.

Animals were immobilized for 24 hours. Indomethacin was applied simultaneously with the immobilization and 12 hours later. After 24 hours, the animals were sacrificed.

Induction of bony, wound, skin and mucosal burns Tibial fractures were created by finger pressure using a hypomechlion under light ether anesthesia. The animals were sacrificed 5, 8, 9, 12, and 30 days after the formation of the fracture.

Large skin wounds were formed under light ether anesthesia. Each animal is dosed daily for the first 5 days, and then
The animals were sacrificed on a 15-day cycle from day 15 and at the end of the 5-day cycle one month after wounding.

Under light ether anesthesia, a scald was formed on the animal's back (15 × 30 mm) for 15 seconds with a cautery cautery.
Each animal was sacrificed 5, 9, and 30 days after lesion formation.

Both nasal mucosa were burned for 5 seconds with a cauterizer under light ether anesthesia. Each animal was sacrificed 5 and 30 days after lesion formation.

Formation of Kidney Injury Ligation of the kidney stem was performed under light ether anesthesia.
Each animal was sacrificed after 24 hours. The wound was sutured and the animals were released freely until death or lethality was reached.

Induction of anti-inflammatory, anti-edema and anti-pyrogenic effects Carrageenan (1% solution) was applied to the forelimbs as a proinflammatory agent (0.5 ml, sc). The limb volume was measured after 3 hours by standard operation.

Siccoferment was used to raise body temperature (2.5g
/ kg, ip). Thereafter, body temperature was measured in the rectum for 3 hours.

Induction of heart failure Each animal was immobilized for 24 hours. Indomethacin was applied at the start of immobilization (0 hours) and 12 hours later. After 24 hours, the animals were sacrificed.

Apply isoprenaline at time 0 and 24 hours (30.0
mg / kg, ip) and the animals were sacrificed after 48 hours.

Induces CNS effects-Examination of effects on spontaneous behavior of animals (mouse) To establish the presence of dopamine neurons in the activity of the substance BPC in the CNS, the usual dopamine agonists apomorphine (20 mg / kg, sc) and cetronin (Five
mg / kg, ip) (comparative control) or saline (0.9% NaCl)
(Control) was administered. The behavior of the animals after the administration was examined. The paradynamic behavior was evaluated according to the method of Malick (1983).

-Examination of effect on iris muscle The test solution was applied by local or intraperitoneal administration. The pupil diameter is determined at appropriate time intervals (5, 10, 15, 30,
60, 90 and 120 minutes).

-Examination of the effect on barbiturate depression activity Anesthesia was induced by application of Na-pentabarbital (50 mg / kg, iv). The duration of barbiturate sleep was assessed by the presence or absence of a so-called "righting reflex".

Induction of effects on diabetes Diabetes was induced by the application of alloxan (100 mg / kg, ip) and the administration of streptozotocin (100 mg / kg, ip). Glucose content in both urine and blood was measured according to standard procedures. Each animal was left free until death.

Estimation of Efficacy Distribution Studies using BPC labeled with I-125 have shown dose- and time-dependent BPC accumulation as in most stomachs, as well as distribution in whole organisms after oral or parenteral application. Show.

Clinical observations The general condition of BPC-treated animals in studies treating liver, pancreas, stomach and duodenum, kidney, heart damage, fractures, wounds, burns (skin, mucous membranes), and diabetes is more significant than in the control group Was good. Unlike the control group, there were no deaths in the treated group,
Alternatively, low mortality was consistently observed.

Microscopic and pathological observations Large yellow-green hepatic necrosis develops in animals of the control group with ligation of the bile duct and hepatic arteries, and in some cases whole lobule and pancreatic necrosis and bleeding damage in the stomach and duodenum Was included. The application of BPC successfully prevented the above damage in a dose-dependent manner. In the control group animals these lesions appeared 6 hours after ligation,
In the treated animals, on the other hand, they were not visible 48 hours after ligation. History studies also supported the macroscopically observed changes and differences.

Significant necrosis in the pancreas and hemorrhagic damage in the stomach and duodenum occurred in animals in the control group that had the bile duct ligated at the entrance to the duodenum. The administration of BPC prevented the development of the damage described above in a dose-dependent manner. History studies also supported the macroscopically observed changes and differences.

Significant gastric damage was shown in animals from the control group that received water immersion stress, a combination of restraint stress and indomethacin application, and a short (15 minute) pyloric ligation. Administration of BPC significantly reduced (or completely eliminated) the development of gastric damage due to the adverse treatment described above.

Serious kidney damage was found in the animals of the control group that had undergone renal cartilage ligation. The lesions were significantly reduced in BPC-treated rats, and history studies strongly supported the macroscopically observed changes and differences.

BP compared to control animals with tibial fracture
In C-treated animals, the development of hematomas after injury was greatly reduced with early functional use of the injured limb. Quantitative medical history studies using conventional methods showed a rapid rate of recovery callus formation in rats treated during all observation periods.

In the wounded control group animals, a significant development of plasma purulent granulation was observed on the first day after the wound was formed. Unlike those in the control group, BPC-treated rats were non-leaching, dry, and had no visible granulation. History studies supported the macroscopically observed changes and differences during all experimental periods.

Significant myocardial damage was observed in animals of the control group treated with the application of indomethacin to restraint stress, together with high levels of CPK in the adult. Administration of BPC significantly reduced myocardial damage and reduced CPK levels. History studies confirmed macroscopically observed changes and differences.

In animals with experimentally induced diabetes, BP
C abolished transient diabetes in the group treated with alloxin. BP in animals treated with streptozotocin
Administration of C increased survival by almost three-fold. Polyphagia and polydipsia were not significant in all animals treated with BPC.

Anti-inflammatory, anti-edema and anti-pyretic effects In animals treated with BPC, there was a marked reduction in carrageenan-induced edema over the control period compared to the control group.

In animals treated with siccoferment, administration of the BPC substance resulted in a large decrease in body temperature increase compared to the results seen in the control group.

In vitro data The substance BPC was added two days after the start of the culture of human fibroblasts L-929 and melanoma cells B-16. Three days later, the number of cells was evaluated using a cell counter. The substance BPC showed a dose-dependent toxic effect on melanoma cells. No effect was shown on fibroblasts.

There was a marked increase in crust development in treated animals compared to control group animals that had received skin burns. In the control group animals that had formed a mucosal (ie nasal mucosal) burn, significant edema of the damaged tissue was observed during the first two days, followed by plasma purulent secretions and portions of nasal respiration. Functional failure has occurred. No such changes were observed in animals treated with BPC. History studies strongly confirmed macroscopically observed changes and differences.

Effect on CNS The effect of this substance on CNS was examined by observing the behavior of animals (mouse). An increase in locomotion was observed 5 minutes after application. The non-disruptive motility of the animals increased every minute and reached its peak after 30 minutes. During this time, the animals stood upright on their hind limbs, like a candle, and sniffed. Strub reduction was evident, each animal ran around the cage, and the pylorus upright. After applying apomorphine, the changes appeared after 45 minutes and were accompanied by a partial upright in the hind limbs. Only a brief period of hyperactivity was observed after serotonin application (increasing locomotion, weaker sniffing and cleaning).

Application locally (or intraperitoneally) to the eyes resulted in miosis (about 20%).

Application of BPC shortened the time of barbiturate-induced sleep, which was established by a reduction in time without righting reflex (at least 60%).

Effects on blood pressure, intestinal motility, basal body temperature, diuresis epinephrine, noradrenaline, izoprenaline,
The application of BPC has no effect on acetylcholine and serotonin-induced changes in blood pressure, and has no effect on basal blood pressure values.

The application of BPC has no effect on intestinal smooth muscle in vitro (guinea pig).

 Application of BPC did not reduce basal body temperature.

 Application of BPC did not result in increased diuresis.

Mechanism of action The effect of BPC, like many other exogenous proteins, is the result of several mechanisms: direct interaction of the protein or its fragments with cellular states, biological activity (stimulation). Stimulation or inhibition of release by a protein or non-protein receptor or non-receptor linked mechanism, interaction with enzymes that degrade endogenous biologically active peptides A number of protective effects against irrelevant pathological situations may be explained when they are described in common terms-stress (noxis) caused in vivo. That is, the substance BPC exhibits a first adaptation (starting from the stomach) to stressful phenomena that endanger all its organisms. Therefore, the mechanism of action of BPC is
Represents the primary defense mechanism against different stress attacks (Noxis).

From the above, it is concluded that the application of BPC can have a favorable medical history on the course following the disease.

I 1. Acute hepatitis (irrespective of etiology, viral, drug-induced, toxic, fulminant) 2. Chronic hepatitis 3. Cirrhosis 4. Cholestasis 5. Cholelithiasis 6. Cholangitis 7. Liver transplantation-before and after transplantation Treatment 8. Pre- and post-operative treatment, including other hepatic surgical procedures (eg, congenital malformations, benign and malignant bile duct stenosis) II 9. Prevent pancreatitis 10. Improve the course of pancreatitis 11. Postoperative pancreatitis 12. Prevention of complications and death from acute pancreatitis 13. Decrease in frequency of chronic pancreatitis III 14. Acute bleeding in the upper gastrointestinal tract 15. Hot gastritis 16. Acute ulcers in both stomach and duodenum ( 17. Chronic peptic ulcer 18. Non-peptic ulcer of the gastrointestinal tract IV 19. Shock (regardless of etiology) V 20. Protection of renal parenchyma from the development of acute and chronic ischemic and hemorrhagic necrosis 21 Acute and chronic kidney damage and all the factors that cause them VI 22. Bone damage (fractures) 23. Bone treatment Placement (osteotomy) 24. Bone connection (calems) 25. False joint 26. Joint fixation 27. Wound (increased treatment speed) 28. Burn (skin and mucous membrane, increased treatment speed) VII 29. Antitumor activity VIII 30. Anti-inflammatory, anti-edema, anti-rheumatic activity 31. Antipyretic activity 32. Rheumatoid arthritis 33. Osteoarthritis 34. Ankylosing spondylolysis 35. Uric acid arthritis 36. Extra-articular rheumatism (synovitis, tendinitis, Synovitis, humeral scapular periarthritis) 37. Inflammation and edema after surgical and non-surgical treatments, fractures and dislocations IX 38. Acute myocardial infarction 39. Myocarditis 40. Delayed ischemic changes in tissue X 41. Low food consumption XI 42. Hyperprolactinemia XII 43. Parkinson's intractable disease, Parkinsonism XIII 44. Glaucoma (reduced intraocular pressure-dopamine-like and hirocarpine-like activity) XIV 45. Diabetes XV 46. Barbiturate poisoning XVI 47. All adverse effects of stress and the consequences of stress Can be applied orally or parenterally to pills, capsules, dragees, lingeriets, suppositories, vaginal inserts, injections, nasal inhalants, ointments, creams, gels, sprays, eye drops, ear drops, nasal drops In the form of

Examples of techniques Example 1: Method of separating BPC material The material from which this material is to be separated was obtained repeatedly and reproducibly from the gastric juices of 542 patients and from various animal sources.

This material was stored at 2-8 ° C. This material was homogenized. This is then centrifuged (12000 ^Gg / 30 min). Thereafter, the supernatant is collected. 200 ml of supernatant is added to water (+
Dialysate 1: 125 (4 ° C.), ie, reach a conductivity of 100 μS. Thereafter, the dialysate is freeze-dried. The dried residue is dissolved in 10 ml of double distilled water and applied to a chromatography column packed with a denatured DEAE ion exchange gel (40-80 μm). This column is 25 × 25
0 mm. For dilution, use a buffer solution (sodium acetate 0.01 M, pH
Use 5.2). Detection of the fractions is performed using a UV detector in the 2800 nm range. 40-120 ml fractions were collected. The eluate was first concentrated with AQUACID-3 (Calbiochem, USA) and then lyophilized. The dried material is dissolved in 10 ml of 0.2 MB phosphate buffer (pH 7.4) and then SEPHA
The solution was supplied to a second chromatography column (2.5 × 450 mm) packed with CRYL-S-200 at a flow rate of 1.2 / min. 80-2
00 ml fractions were collected.

The solution is dialyzed against distilled water, concentrated with AQUACID-3 and lyophilized. About 200mg of dry white substance
Is obtained.

Identification The BPC material is characterized by the infrared and ultraviolet spectra shown in FIGS.

Based on the pharmacological properties described above, this substance BPC
Can be used for various diseases and disorders, especially when the normal formation of this substance in vivo is insufficient or even absent.

It can also be used to treat the following diseases:

a) all diseases and diseases induced by stress b) cardiovascular diseases: ulcers (of different origin), shock (of different etiology), ischemia and infarction (of different organs) c) inflammation and Edema: Inflammation and edema-forming etiology, hepatitis (acute and chronic), cirrhosis, cholangitis, pancreatitis (different etiologies), gastroduodenitis, nephritis and nephropathy, myocarditis, pericarditis, endocarditis, arthritis , Arthritis (different etiologies), rhinitis, rheumatic diseases (disorders of different etiological combinations), fever, disorders induced by free radicals. D) Injuries and surgery: trauma (preventive and post-traumatic treatment), transplantation, wounding. E) Marignancy (certain tumors) f) Diabetes mellitus g) Diseases of dopamine-induced or similar pathogenesis: obesity, Parkinson's disease, hyperprolactinemia, glaucoma, and barbi Rate-induced sleep (poisoning with barbiturates) h) Nervous system diseases and disorders: Alzheimer's disease, ischemic and traumatic damage of the nervous system, and delayed onset of senile degeneration i) Stomatology: Diseases and disorders of the oral mucosa J) Hematopoietic system: neutropenia, aplastic anemia, bone marrow transplantation of spinal cord displastic syndrome k) Radiation damage: Protection against acute exposure syndrome and radiation damage l) Immunology: hypersensitivity reactions, allergies and transplants m) Fertility: enhancement of fertility, treatment of azoosperm etc. and promotion of milk distribution n) Veterinary use: reduction of mortality in commercial breeding, stunting O virology: antiviral treatment p) other: liver fat, cholestatic, cholelithiasis, acute and chronic renal dysfunction, kidney Toxic damage to the liver and the substance BPC is used as a drug for enteral, topical, rectal and parenteral applications. These drug compositions are formulated by adding various directional or organic auxiliaries. For tablets and sugar-coated tablets, for example, talc, lactose, starch, stearate and the like are added, and for solutions and suspensions, for example, water, alcohols, glycerin and the like are added. For suppositories, for example, natural fats and oils, hardened fats and oils, waxes and polyethylene glycol are added. These preparations may generally contain suitable preservatives, buffers, stabilizers, surfactants, dissolution mediators, sweeteners and dyes.

A suitable daily dose of the substance BPC is 5 to 50 μg / kg of body weight and can be applied in a single dose or in divided doses.

Example 1 Production of BPC from Human Gastric Fluid The raw material used for the production of this substance BPC was obtained in a reproducible manner from the gastric fluid of 542 patients. The collected material must be stored between + 2 ° C and 8 ° C. It was homogenized before use.

An aliquot (200 ml) of this material was first centrifuged (12000 G, 30 minutes) to separate the crude portion into the supernatant. The supernatant was dialyzed against demineralized water at + 4 ° C. until a conductivity of 100 μS was obtained (1: 125) and the solution was lyophilized. The dried residue is distilled water 10ml
And loaded onto a chromatography column (25 × 250 mm) packed with a DEAE-modified ion exchange gel [eg Trisacyrl M-DEAE® IBF].
Sodium acetate buffer (0.01M, pH = 5.2) for elution
Was used. Each fraction was controlled with a 280 nm UV detector.
The eluate fractions from 40 ml to 120 ml were collected and
Concentrate using ID-3 (registered trademark, Colbinochem),
It was then dialyzed and lyophilized. The dried material is dissolved in 10 ml of 0.2 M phosphate buffer (pH = 7.4) and again
hacryl-S-200 (registered trademark, Pharmacia Fine Chemical
(Company s). The flow rate was 1.2 ml / min. The fractions from the 60th to the 200ml were collected, dialyzed against distilled water, and Aquacid-
3 and then lyophilized. 3 mg of a white substance was obtained. The resulting BPC is very soluble in water. It decomposes between 180 ° C and 200 ° C. The molecular weight determined by chromatography is 40,000 ± 5,000
It is. The ultraviolet spectrum shows several ranges of absorption shown in FIG.

The infrared spectrum (in KBr) shows absorption bands in the range shown in FIG. This substance BPC has a partial amino acid sequence from the N-terminus H ₂ N-Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val- ‥‥‥- It has a fold protein structure containing COOH.

Example 2 Production of BPC from Human Gastric Fluid Homogenized human gastric fluid of the same origin as in Example 1
, Dialyzed and ligated and dried.

The dried residue (80.4 mg) was added to a 0.1 M acetate buffer (3 ml, p
H = 5.22), which was previously equilibrated with acetate buffer (pH = 5.2).
It was added to a chromatography column (10 × 50 mm) packed with ogel-TSK (registered trademark, Marck). Flow rate is 0.7ml / mi
n. Each fraction was collected at 280 nm with control by a UV detector. The first fraction with strong UV absorption was collected.

This fraction was dialyzed against distilled water and then lyophilized (47 mg).

The dried residue was redissolved in acetate buffer (1 ml) and the gel was pre-equilibrated with acetate buffer.
Sephadex-G-100 (registered trademark, Pharmacia Fine Chemic)
als) packed chromatography column (16 × 560
mm). The flow rate was 0.3 ml / min. From 66ml eyes 1
Fractions up to 08 ml were collected, dialyzed against distilled water and lyophilized. A white powder was obtained which was consistent with the material described in the previous example.

Example 3 Preparation of the substance BPC from pig gastric juice Homogenized pig gastric juice (85 ml) was centrifuged, dialyzed according to the procedure described in the previous example and lyophilized to give 440 mg of dry residue. Then 110 mg of the dried residue was dissolved in 0.1 M acetate buffer (6 ml, pH = 5.22) and added to a chromatography column (10 × 50 mm) filled with ion exchange resin Fractogel-TSK. , And processed as described in the preceding example. 44mg yellowish concentrate
was gotten.

One part (24 mg) of this concentrate was dissolved in 1 ml of the same acetate buffer and this was filled with gel Sephadex®-50 / 80, which had been equilibrated with acetate buffer. Added to a chromatography column (16 × 565 mm).
The yield was 1.35 ml / min. The eluent from the 65th to 108th ml was collected, dialyzed against distilled water and lyophilized. The resulting white powder (10 mg) has the same molecular weight and other properties as the product described in the preceding columns, within the precision and reproducibility of the analytical method used.

Example 4 Tablet composition mg substance BPC per tablet 0.3 Lactose 37.7 Starch 6.0 tragacanth 2.5 Magnesium stearate 0.5 50 mg Example 5 Capsule composition mg substance BPC per tablet 0.3 Lactose 99.0 Magnesium stearate 0.7 100 mg Example 6 Ringeret Composition mg substance per tablet BPC 0.3 mannitol 100.0 Starch 5.0 polyvinylpyrrolidone 1.5 talc 3.2 110 mg Example 7 Suppository composition mg substance BPC per tablet 0.5 polyethylene glycol 300 478.0 polyethylene glycol 1500 529.0 polyethylene glycol 6000 38.0 emulsifier 14.5 1060 mg Example 8 Cream composition mg substance per tablet BPC 0.05 Emulsifier Emulsifier 25.00 Hydrogenated oleum arachid 50.00 tween 60 120.00 Propylene glycol 1500 30.00 Methylhydroxybenzoate 0.16 Propylhydroxybenzoate 0.17 250.00 mg Example 9 Injection composition Per tablet M g substance BPC 0.3 sodium carboxymethyl cellulose 1.5 polyvinyl pyrrolidone 5.8 lecithin 2.7 benzyl alcohol 0.01 buffer qs double distilled water added 1 ml Example 10 Solution composition mg substance BPC per tablet 0.05 glycerol 15.00 benzyl alcohol 0.01 buffer qs redistillation 25 ml of water added Example 11 Dry Injection Composition mg substance BPC per tablet 0.3 Manirole 10.0 10.3 mg Example 12 Nasal inhalant composition mg substance BPC per tablet 0.5 Benzyl alcohol 0.15 Uliglycol®-812 14.00 Freon 12/114 60.00 74.65mg Example 13 Drop composition for eyes, ears and nose mg substance per tablet BPC 45 Stabilizer 5 Sodium hydrogen phosphate 116 Sodium dihydrogen phosphate 33 Benzalkonium chloride 0.5 Double distilled water Additional 5 ml

 ──────────────────────────────────────────────────続 き Continued on the front page (73) Patent holder 999999999 Mise Stepan Hugoslavia Waiyu 58000 Splittorzwaldva 37 (73) Patent holder 999999999 Krisanak Simun Yugoslavia Waiyu 41000 Zagreb Platana 8 (73) Patent holder 999999999 Udvi チ k Ivan Switzerland Zeha 6370 Stans Nvé Ennet Moserstraße 16 (73) Patent holder 999999999 Schanek Ernest Hugoslavia Waiyu 41000 Zagreb Aleya Ve Popovika 125 (73) Patent holder 999999999 Duvnaq Marco Yugoluvia 4 Luxembourg 4 (73) Patent holder 999999999 Yukiku Jeruka You -Slavia Waiyu 41000 Zagreb Pali Omuradinaka 33 (72) Inventor Shikirik Predrag Yugoslavia Waiyu 41000 Zagreb Yulisicheva 5 (72) Inventor Petek Mariyan Yugoslavia Waiyu 41000 Zagreb Visnikka 29 (72) Waiyu 41000 Zagreb Chuetno Nasaries 1/21 (72) Inventor Mise Stepan Hugoslavia Waiyu 58000 Splits Waldva 37 (72) Inventor Krisanak Simun Yugoslavia Waiyu 41000 Zagreb Platana 8 (72) Inventor Switzerland Udvich Zeha 6370 Stans Nvé Ent Moserstrasse 16 (72) Inventor Schanek Ernest Hugoslavia Waiyu 41000 Zagreb Aleja E over Popovi mosquito 125 (72) inventor Duvunyaku Marco Yugoslavia Waiyuu-41000 Zagreb el Luxembourg 4 (72) inventor Yukiku Ieruka Yugoslavia Waiyuu-41000 Zagreb Parry Omuradinaka 33

Claims (27)

(57) [Claims] In producing a substance BPC from human or animal gastric juice, the following production steps are performed: a) The collected and homogenized animal or human gastric juice is collected and centrifuged to obtain a crude part and a supernatant. B) dialyzing the supernatant and freeze-drying; c) purifying the dried residue on a chromatography column packed with a weakly basic modified ion-exchange gel or resin. D) The concentrate is dialyzed again and freeze-dried, e) The dried residue is purified by gel chromatography, and f) The concentrate is measured by gel chromatography using dialysis and freeze-drying. Molecular weight 40,000 ± 5,000
0 Dalton, infrared spectrum and ultraviolet spectrum shown in FIG. 1 and FIG. 2 and partial amino acid sequence of N-terminal H ₂ N-Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp A method characterized by finally obtaining a substance BPC having -Ala-Gly-Leu-Val- ‥‥‥ -COOH. 2. The process according to claim 1, wherein in step c) a DEAE, DMAE, QAE or other basic group modified anion exchange gel or resin is used. 3. In step e), use is made of an inorganic or organic gel having a dextran, polystyrene, polyacrylamide, agarol, polyvinyl or other matrix having a molecular weight ranging from 20,000 to 100,000 daltons. 3. The method of claim 1 or claim 2. 4. A molecular weight of 40,000 ± 5,000 daltons as determined by gel chromatography, an infrared spectrum and an ultraviolet spectrum shown in FIGS. 1 and 2, and an N-terminal partial amino acid sequence H ₂ N-Gly-Glu-Pro- A substance BPC having Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val- ‥‥‥ -COOH. 5. The molecular weight of 40,000 ± 5,000 daltons as determined by gel chromatography, the infrared and ultraviolet spectra shown in FIGS. 1 and 2 and the partial amino acid sequence at the N-terminus H ₂ N-Gly-Glu-Pro- A substance BPC having Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val- ‥‥‥ -COOH and obtained by any one of the above claims 1 to 3. And the substance BP used to treat ulcers
C. 6. The substance BPC according to claim 5, which is used for treating myocardial infarction instead of treating ulcer. 7. The substance BPC according to claim 5, which is used for treating a damaged liver instead of treating an ulcer. 8. The substance BPC according to claim 5, which is used for treating a damaged pancreas instead of treating an ulcer. 9. The substance BPC according to claim 5, which is used for treating a damaged kidney instead of treating an ulcer. 10. The substance BPC according to claim 5, which is used for treating diabetes mellitus instead of treating ulcer. 11. The substance BPC according to claim 5, which is used for treating fever instead of treating ulcer. 12. The substance BPC according to claim 5, which is used for treating edema instead of treating ulcer. 13. The substance BPC according to claim 5, which is used for treating rhinitis instead of treating ulcers. 14. The substance BPC according to claim 5, which is used for treating a fracture instead of treating an ulcer. 15. The substance BPC according to claim 5, which is used for treating burns instead of treating ulcers. 16. The substance BPC according to claim 5, which is used for treating skin wounds instead of treating ulcers. 17. The substance BPC according to claim 5, which is used for treating bowel disorders instead of treating ulcers. 18. The substance BPC according to claim 5, which is used for treating a tumor instead of treating an ulcer. 19. The substance BPC according to claim 5, which is used for treating radiation damage instead of treating ulcers. 20. The substance BPC according to claim 5, which is used for treating hematopoietic disorders instead of treating ulcers. 21. The substance BPC according to claim 5, which is used for treating azoospermia instead of treating ulcer. 22. The substance BPC according to claim 5, which is used for improving reproductive function instead of treating ulcers. 23. The substance BPC according to claim 5, which is used for treating periodontal tissue damage instead of treating ulcers. 24. The substance BPC according to claim 5, which is used for treating damage to basal ganglia neurons instead of treating ulcers. 25. The substance BPC according to claim 5, which is used for treating spinal cord contusion instead of treating ulcer. 26. The substance BPC according to claim 5, which is used for improving commercial breeding instead of treating ulcers. 27. The substance BPC according to claim 5, which is used for improving antiviral activity instead of treating ulcers.