EP1567128A1 - Process for obtaining bacterium-size particles - Google Patents
Process for obtaining bacterium-size particlesInfo
- Publication number
- EP1567128A1 EP1567128A1 EP03701627A EP03701627A EP1567128A1 EP 1567128 A1 EP1567128 A1 EP 1567128A1 EP 03701627 A EP03701627 A EP 03701627A EP 03701627 A EP03701627 A EP 03701627A EP 1567128 A1 EP1567128 A1 EP 1567128A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- cholesterol
- particles
- changing
- solution
- size
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- 239000002245 particle Substances 0.000 title claims abstract description 69
- 238000000034 method Methods 0.000 title claims abstract description 43
- 230000008569 process Effects 0.000 title claims abstract description 38
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 173
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 75
- 230000003627 anti-cholesterol Effects 0.000 claims abstract description 25
- 239000012528 membrane Substances 0.000 claims abstract description 25
- 239000000243 solution Substances 0.000 claims abstract description 25
- 238000004519 manufacturing process Methods 0.000 claims abstract description 18
- 239000013078 crystal Substances 0.000 claims abstract description 13
- 238000002649 immunization Methods 0.000 claims abstract description 10
- 239000012047 saturated solution Substances 0.000 claims abstract description 10
- 241000192041 Micrococcus Species 0.000 claims abstract description 9
- 230000003053 immunization Effects 0.000 claims abstract description 9
- 241000894006 Bacteria Species 0.000 claims abstract description 7
- 150000001841 cholesterols Chemical class 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000012454 non-polar solvent Substances 0.000 claims abstract description 4
- 239000002798 polar solvent Substances 0.000 claims abstract description 4
- 210000004369 blood Anatomy 0.000 claims description 16
- 239000008280 blood Substances 0.000 claims description 16
- 239000003960 organic solvent Substances 0.000 claims description 10
- 230000001965 increasing effect Effects 0.000 claims description 9
- 241000282414 Homo sapiens Species 0.000 claims description 7
- 241001465754 Metazoa Species 0.000 claims description 5
- 230000007812 deficiency Effects 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 230000001939 inductive effect Effects 0.000 claims description 5
- 238000010253 intravenous injection Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 230000007423 decrease Effects 0.000 claims description 4
- 239000011368 organic material Substances 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 239000008151 electrolyte solution Substances 0.000 claims description 3
- 229910010272 inorganic material Inorganic materials 0.000 claims description 2
- 239000011147 inorganic material Substances 0.000 claims description 2
- 230000000717 retained effect Effects 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- VMXUWOKSQNHOCA-UKTHLTGXSA-N ranitidine Chemical compound [O-][N+](=O)\C=C(/NC)NCCSCC1=CC=C(CN(C)C)O1 VMXUWOKSQNHOCA-UKTHLTGXSA-N 0.000 claims 1
- 230000000638 stimulation Effects 0.000 abstract description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 7
- 230000004060 metabolic process Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 230000000890 antigenic effect Effects 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 239000002502 liposome Substances 0.000 description 5
- 230000002052 anaphylactic effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 208000035150 Hypercholesterolemia Diseases 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000004962 physiological condition Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000002009 allergenic effect Effects 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 235000018823 dietary intake Nutrition 0.000 description 2
- 230000005670 electromagnetic radiation Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 208000006379 syphilis Diseases 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 206010002199 Anaphylactic shock Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 238000008214 LDL Cholesterol Methods 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000002546 agglutinic effect Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000024203 complement activation Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 208000035474 group of disease Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- YWXYYJSYQOXTPL-SLPGGIOYSA-N isosorbide mononitrate Chemical compound [O-][N+](=O)O[C@@H]1CO[C@@H]2[C@@H](O)CO[C@@H]21 YWXYYJSYQOXTPL-SLPGGIOYSA-N 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1277—Processes for preparing; Proliposomes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1274—Non-vesicle bilayer structures, e.g. liquid crystals, tubules, cubic phases, cochleates; Sponge phases
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
- A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids
Definitions
- the invention is a process for producing particles of bacterium size from cholesterol, as well as the particles made by the process suitable for stimulation of production of anticholesterol antibody and the process for producing micrococcus size (100-500 nm) particles suitable for immunization with the use of cholesterol of high purity degree.
- the invention relates to a process, which is suitable for producing micrococcus size particles from cholesterol of high purity degree without the use of other lipoids and on the surface of the said particles a membrane containing only high purity cholesterol can be found.
- Atherosclerosis is thought to develop due to high cholesterol level in blood .
- Both methods can only ameliorate symptoms, but can not solve the problem of deficiency of cholesterol metabolism. Removal of cholesterol from the blood stream still presents a problem.
- Cholesterol is not antigenic, or it is only antigenic to a small extent. A substance should be produced from cholesterol, which can stimulate production of anticholesterol antibodies.
- Drug therapy is used for preventing development of excess cholesterol. Absorption of cholesterol from dietary intake is prevented by transforming cholesterol into a complex substance in the intestines that can not be absorbed. Production of cholesterol (endogenous) arising within the body can be controlled by hindering functioning of enzymes taking part in cholesterol synthesis. Both solutions intervenes in cholesterol metabolism by preventing the body to get cholesterol. However cholesterol is absolutely necessary for the organism under physiological conditions (bile acids, hormones, cell membranes, receptor stabilizers) therefore it is not the best solution to prevent cholesterol from the body, because cholesterol deficiency can also cause various diseases.
- the invention relates to a process for producing particles of bacterium size from cholesterol during said process a supersaturated solution is heated to the level of saturation, and a nearly saturated solution is made, which is characterized by that, conditions for producing liquid membranes arranged as crystals from cholesterol molecules are created by changing the cholesterol in the unsaturated cholesterol solution with continuous, equal changing of the combination ratio of polar and non- polar solvents, then by transmitting physical energy of determined amplitude and energy said membranes are transformed to particles having a characteristic structure that can be observed by a microscope.
- ordinary, common cholesterol in trade is specially purified for the process, during said process a saturated solution is produced in an organic solvent at 56-90 °C, -then said solution is crystallized by changing the temperature, then crystals are collected on a sterile filter, dried and said crystalls are solved in another solvent and a saturated solution is produced at 56-90 °C then it is crystallized by changing the temperature,
- the invention relates further to particles produced preferably by the process according to the invention, which is characterized by that, the surface membrane of the particle of micrococcus size constitutes of cholesterol of high purity degree.
- the structure of the membrane is retained stable in an electrolytic suspension or in a lyophilized form .
- the particle encapsules part of different size, preferably organic materials of great molecules and inorganic materials jointly or separately as well.
- the particle in itself, or filled with electrolytic solutions or with organic materials of great molecules is applied for active and immune-stimulation and active immunization.
- the particles according to the invention are suitable for intravenous injection into any animal (human being) whose blood can contain cholesterol among natural circumstances.
- 'vacant' particles are applied for inducing production of anticholesterol antibody.
- antibodies formed under influence of particles are suitable for prevention and treatment of diseases contracted due to deficiency of anticholesterol antibody or due to decline of antibody titre.
- the monomolecular parts of membrane arranged by the oscillator and torn to films in the ultra micro grooves are entwined to globular formulas of 100-500 nm diameter by electromagnetic radiation.
- the bacterium-like particles produced by the above process are collected in a physiological suspension under sterile conditions and are stored in the form of an 1 mg/ml suspension calculated on the dry-material content of the cholesterol at 5°C in one ml doses or in bigger volume, which can be dozed before use as required.
- Crystals are collected on a sterile filter, dried, then another saturated solution is prepared in another organic solvent at 56-90 °C.
- Crystals are collected under sterile conditions, dissolved in an organic solvent at 56-90 °C, and are stored as a supersaturated solution at 5 °C.
- Particles produced according to the process according to the invention can be characterized by that,
- Particles can be dried by lyophilization, and can be resuspended.
- the advantage of the process according to the invention is, that it makes possible the production of particles with immunizing characteristics for the organism in a way, that pure cholesterol can function as a particle of bacterium size, which stimulates production of antibody involved in cholesterol metabolism in the organism.
- the novelty of our process is, that it makes possible the production of a monomolecular membrane from pure cholesterol.
- the advantage of the particles produced from pure cholesterol compared to liposomes produced from the mixture of several lipoids is, that they induce increase of titre of anticholesterol antibody in the organism without creating allergenic affects creating anaphylactic effects.
- the particles produced from pure cholesterol are suitable for intravenous injection into any organism in a concentration of 1 mg/ml in a dose of 9 x 1 ml.
- particles of micrococcus size from cholesterol of high purity degree, which are suitable for inducing production of anticholesterol antibody in the form in intravenous injection by active immunization and are suitable for treatment of diseases contracted due to deficiency of anticholesterol antibody or due to decline of anticholesterol antibody titre.
- the stabilizing role is ensured by well-known additives and additives ensuring isotonic conditions.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Dispersion Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention is a process for producing particles of bacterium size from cholesterol, as well as the particles made by the process suitable for stimulation of production of anticholesterol antibody and the process for producing micrococcus size (100-500 nm) particles suitable for immunization with the use of cholesterol of high purity degree. The invention relates to a process for producing particles of bacterium size from cholesterol during said process a supersaturated solution is heated to the level of saturation, and a nearly saturated solution is made, which is characterized by that, conditions for producing liquid membranes arranged as crystals from cholesterol molecules are created by changing the cholesterol in the unsaturated cholesterol solution with continuous, equal changing of the combination ratio of polar and nonpolar solvents, then by transmitting physical energy of determined amplitude and energy said membranes are transformed to particles having a characteristic structure that can be observed by a microscope. The invention relates further to particles produced preferably by the process according to the invention, which is characterized by that, the surface membrane of the particle of micrococcus size constitutes of cholesterol of high purity degree.
Description
Process for obtaining bacterium-size particles
The invention is a process for producing particles of bacterium size from cholesterol, as well as the particles made by the process suitable for stimulation of production of anticholesterol antibody and the process for producing micrococcus size (100-500 nm) particles suitable for immunization with the use of cholesterol of high purity degree. The invention relates to a process, which is suitable for producing micrococcus size particles from cholesterol of high purity degree without the use of other lipoids and on the surface of the said particles a membrane containing only high purity cholesterol can be found.
In the state of the art there is no effective process for preventing atherosclerosis and there is no really effective medication for curing this disease either. Consequently atherosclerosis seems to belong to the group of diseases incurable or curable to a limited extent only.
Atherosclerosis is thought to develop due to high cholesterol level in blood .
Therefore treatment was focused on decreasing cholesterol level of blood.
So far two methods have been applied for treating high cholesterol level of blood:
- prevention of absorption of cholesterol from the intestines
- prevention of production of endogenous cholesterol
Both methods can only ameliorate symptoms, but can not solve the problem of deficiency of cholesterol metabolism. Removal of cholesterol from the blood stream still presents a problem.
Cholesterol is not antigenic, or it is only antigenic to a small extent. A substance should be produced from cholesterol, which can stimulate production of anticholesterol antibodies.
During earlier research an antibody was observed in the human body, that reacted with cholesterol. (Horvath, I. Puskas, E. Horvath, Anna Balazs, E. Horvath, A: Autoantibody or immunological defense against cholesterol Orvosi Hetilap Medical Weekly 1994. 135.18.965-968). Observation of antibodies resulted from the experiences gained during our examinations for the verification of lues when we could detect an antibody, an anticholesterol, reacting with cholesterol in seroreactive serums of non-lues origin.
Searching the biological function of antibody we studied the characteristics of the antibody with various cholesterol substances. (Horvath, Anna Varga, L. Horvath, I. Kόkai, M. Karadi, I. Romics, L. Fust, G.: Complement activation in human sera by liposomes made from cholesterol, Mol.Immunology, 1998, 35(6-7): 102 (IF: 1,824) (Horvath, I. Szende, B. Horvath, Anna Kocsis, I. Horvath, A. : Influence of cholesterol liposome immunisation and immunstimulation on rabbits atherosclerosis induced by a high cholesterol diet Med. Sci.Monit. 1998. 4(3):403-407).
Later we succeeded in producing a composition by a method elaborated and stabilized by us inducing production of antibody on experimental animals with great certainty. During the process we produced cholesterol particles suitable for immunization which proved to be of antigenic character and were suitable for inducing production of anticholesterol antibodies. Anticholesterol antibody produced this way considerably dicreased the cholesterol level of blood previously increased by a cholesterol diet and protected the experimental animals from atherosclerosis.
During human tests a higher titre of anticholesterol antibody was observed in a sound constitution compared with organism of patients of atherosclerosis of the same age and gender. (Horvath, Anna Prohaszka, Z. Duba, J. Harczos, P. Horvath, I. Vallus, G. Romics, L. Fust, Gy. Karadi, I: Comperative examination of anticholesterol antibody titre in patiens of atherosclerosis Magyar Belorvosi Archivum Archives of Hungarian Internists 1998/3:239) The titre values of anticholesterol antibody and change of cholesterol level of blood in animal tests showed similarity with values measured in human beings. We may conclude from this the fact, that anticholesterol antibody plays an important role in cholesterol metabolism of the human organism as well.
The solutions in the state of the art suffer limitations due to the following facts: The principal causes of high cholesterol level in blood are as follows:
- dietary intake rich in animal fats
- cholesterol developed by the organism
Drug therapy is used for preventing development of excess cholesterol. Absorption of cholesterol from dietary intake is prevented by transforming cholesterol into a complex substance in the intestines that can not be absorbed. Production of cholesterol (endogenous) arising within the body can be controlled by hindering functioning of enzymes taking part in cholesterol synthesis. Both solutions intervenes in cholesterol metabolism by preventing the body to get cholesterol. However cholesterol is absolutely necessary for the organism under physiological conditions (bile acids, hormones, cell membranes, receptor stabilizers) therefore it is not the best solution to prevent cholesterol from the body, because cholesterol deficiency can also cause various diseases.
According to data of textbooks it is not possible to produce liposome from pure cholesterol, because pure cholesterol becomes chrystalline in watery substance and chrystals prevent the formation of a lipoid membrane. So far it was not possible to create a membrane from cholesterol with the methods applied, which membrane would have been suitable in itself for creating a monomolecular membrane in a watery substance.
Recently a few research groups realized the important role of anticholesterol antibody in the cholesterol metabolism. In vitro tests proved, that anticholesterol antibody is linked mainly to the LDL cholesterol. During in vivo tests the cholesterol level of blood could be considerably decreased by increasing the titre of the anticholesterol antibody. This solution achieved the decrease of the cholesterol
level of blood in a way, that cholesterol necessary for the body was not removed. Antigenes increasing anticholesterol production created by certain researches have caused anaphylactic shock in different animals.
When elaborating the solution according to the invention we aimed to produce a process as well as a particle produced by the said process, which induce increase of titre of anticholesterol antibody in the organism without creating allergenic affect of the antigen.
When working out the solution according to the invention we realized, that in case we produce particles of antigen character from pure cholesterol, then no anaphylactic affect will show, beside the favourable affect of immunisation. We produced various cholesterol antigenes and examined their effects. We found, that particles of bacterium size can be produced from the membrane made from pure cholesterol. Despite the fact, that such particles contain cholesterol only, they have increased immunological features.
The invention relates to a process for producing particles of bacterium size from cholesterol during said process a supersaturated solution is heated to the level of saturation, and a nearly saturated solution is made, which is characterized by that, conditions for producing liquid membranes arranged as crystals from cholesterol molecules are created by changing the cholesterol in the unsaturated cholesterol solution with continuous, equal changing of the combination ratio of polar and non- polar solvents, then by transmitting physical energy of determined amplitude and energy said membranes are transformed to particles having a characteristic structure that can be observed by a microscope.
One of the preferred applications of the process according to the invention, in order to establish stabile particles said particles are treated for 1 to 10 minutes changing the ratio of different solutions equally and continuously, increasing frequency of physical impact many times, then they are dosed to the required volumes after control by a dark field microscope.
In another of the preferred applications of the process according to the invention, ordinary, common cholesterol in trade is specially purified for the process, during said process a saturated solution is produced in an organic solvent at 56-90 °C, -then said solution is crystallized by changing the temperature, then crystals are collected on a sterile filter, dried and said crystalls are solved in another solvent and a saturated solution is produced at 56-90 °C then it is crystallized by changing the temperature,
- then collected on another sterile filter the crystals are dissolved in a third organic solvent at 56-90 °C, and are crystallized again by changing the temperature,
- crystals are collected under sterile conditions, dissolved in an organic solvent at 56-90 °C, and are stored as a supersaturated solution at 5 °C.
The invention relates further to particles produced preferably by the process according to the invention, which is characterized by that, the surface membrane of the particle of micrococcus size constitutes of cholesterol of high purity degree.
One of the preferred realization of the particles according to the invention, the structure of the membrane is retained stable in an electrolytic suspension or in a lyophilized form .
In another preferred realization of the particles according to the invention, the particle encapsules part of different size, preferably organic materials of great molecules and inorganic materials jointly or separately as well.
In a further preferred realization of the particles according to the invention, the particle in itself, or filled with electrolytic solutions or with organic materials of great molecules is applied for active and immune-stimulation and active immunization.
In a further preferred realization of the particles according to the invention, they are suitable for intravenous injection into any animal (human being) whose blood can contain cholesterol among natural circumstances.
In a further preferred realization of the particles according to the invention, 'vacant' particles are applied for inducing production of anticholesterol antibody.
In a further preferred realization of the particles according to the invention, antibodies formed under influence of particles are suitable for prevention and treatment of diseases contracted due to deficiency of anticholesterol antibody or due to decline of antibody titre.
The solution according to the invention is set forth in the following examples:
Example 1
Production of cholesterol particles
20 grams of cholesterol is solved in 1000 ml ethanol and under pressure it is spread with a speed of lOOOul sec on the 15 ° angle surface dented trough-like of metal sheet of determined silicium content or an acid-proof metal sheet or on the surface of a sheet formed as a pipe, which has equal ultramicro grooves parallel with the axis of the pipe. The three-dimensional sizes of the grooves of the surface parallel with the axis of the trough are controlled by a MULTISCOPE. The depth and width of grooves can be 100-1000 nm. The sheet receiving the solution is exposed to physical impact with the help of a tuned oscillator and the surface is treated with an electromagnetic radiation of 270-1080 nm wavelength from a tuned monochromatic power-source during flowing of the fluid.
The monomolecular parts of membrane arranged by the oscillator and torn to films in the ultra micro grooves are entwined to globular formulas of 100-500 nm diameter by electromagnetic radiation. The bacterium-like particles produced by the above process are collected in a physiological suspension under sterile conditions and are stored in the form of an 1 mg/ml suspension calculated on the dry-material content of the cholesterol at 5°C in one ml doses or in bigger volume, which can be dozed before use as required.
Example 2
Purification of cholesterol:
Ordinary, common cholesterol in trade is specially purified for the process as follows:
- l.We produce a saturated solution in an organic solvent at 56-90 °C, then it is crystallized by changing the temperature.
- 2. Crystals are collected on a sterile filter, dried, then another saturated solution is prepared in another organic solvent at 56-90 °C.
- 3. It is crystallized by changing the temperature, then collected on an other sterile filter the crystals are dissolved in a third organic solvent at 56-90 °C, and are crystallized again by changing the temperature.
- 4. Crystals are collected under sterile conditions, dissolved in an organic solvent at 56-90 °C, and are stored as a supersaturated solution at 5 °C.
Example 3
Production of the particles
Supersaturated solution is heated to the level of saturation, then a nearly saturated solution is made.
1. We create conditions for producing liquid membranes arranged as crystals from cholesterol molecules by changing the cholesterol in the unsaturated cholesterol solution with continuous, equal changing of the combination ratio of polar and non- polar solvents. Transmitting physical energy of determined amplitude and energy they are transformed to particles having a characteristic structure that can be observed by a microscope.
2. In order to establish stabile particles they are treated for 1 to 10 minutes changing the ratio of different solutions equally and continuously, increasing frequency of physical impact many times, then they are dosed to the required volumes after control by a dark field microscope .
Particles produced according to the process according to the invention can be characterized by that,
- 1. Their outer appearance on an ultramicroscope is similar to Micrococcus.
- 2. They settle relatively quickly when suspended in electrolytic solutions, and they retain their characteristics for years even at room temperature.
- 3. Particles can be dried by lyophilization, and can be resuspended.
- 4. They tend to agglutination.
- 5. Agglutinative features are decreased or eliminated in solutions of albumin and globulin content.
The advantage of the process according to the invention is, that it makes possible the production of particles with immunizing characteristics for the organism in a way, that pure cholesterol can function as a particle of bacterium size, which stimulates production of antibody involved in cholesterol metabolism in the organism. The novelty of our process is, that it makes possible the production of a monomolecular membrane from pure cholesterol.
The advantages of the particles produced by process according to the invention, preferable applications:
The advantage of the particles produced from pure cholesterol compared to liposomes produced from the mixture of several lipoids is, that they induce increase of titre of anticholesterol antibody in the organism without creating allergenic affects creating anaphylactic effects. The particles produced from pure cholesterol are suitable for intravenous injection into any organism in a concentration of 1 mg/ml in a dose of 9 x 1 ml.
The advantage of this procedure is, that particles getting to the blood can reach lymphocytes in the blood-stream responsible for immunity directly therefore a favourable effect can be achieved by a relatively small dose. Membranes produced from lipoids and various particles (liposomes) produced from the membranes generally have lipoid ratios under physiological conditions. The advantage of these membranes is, that their production is simple, their disadvantage is, that they may create anaphylactic effects during immunization.
Production of membranes from pure cholesterol can be regarded a novelty compared to the processes in the state of the art, because no pure cholesterol membrane is produced in the organism under physiological conditions. In general cholesterol becomes chrystals in watery substance, or occurs as a complex with proteins. Particles produced from pure cholesterol can be well suspended in watery substance, they are stabile, have increased antigenic features and do not create anaphylactic effects.
We worked out a process according to the invention, resulting in a normalization of the cholesterol level of blood in a way, that cholesterol of 'bad' quality in the blood gets involved in the cholesterol metabolism. The gist of it is, that the level of anticholesterol antibody is increased in the blood by immunization of the cholesterol made antigenic on the surface of the particles of micrococcus size. Due to the higher titre of anticholesterol antibody the surface of LDL is coated with protein, antibody and becomes linkable for the receptors of the liver. This way excess of cholesterol can be removed from the blood and the excess of cholesterol can be even used. It results in recovering of the broken chain of cholesterol metabolism.
It is possible to produce particles of micrococcus size from cholesterol of high purity degree, which are suitable for inducing production of anticholesterol antibody in the form in intravenous injection by active immunization and are suitable for treatment of diseases contracted due to deficiency of anticholesterol antibody or due to decline of anticholesterol antibody titre. In case of intravenous injection of the particle suspension the stabilizing role is ensured by well-known additives and additives ensuring isotonic conditions.
Claims
1. Process for producing particles of bacterium size from cholesterol during said process a supersaturated solution is heated to the level of saturation, and a nearly saturated solution is made, characterized by that, conditions for producing liquid membranes arranged as crystals from cholesterol molecules are created by changing the cholesterol in the unsaturated cholesterol solution with continuous, equal changing of the combination ratio of polar and non- polar solvents, then by transmitting physical energy of determined amplitude and energy said membranes are transformed to particles having a characteristic structure that can be observed by a microscope.
2. Process according to claim 1, characterized by that, in order to establish stabile particles said particles are treated for 1 to 10 minutes changing the ratio of different solutions equally and continuously, increasing frequency of physical impact many times, then they are dosed to the required volumes after control by a dark field microscope.
3. Process according to claims 1 or 2 characterized by that, ordinary, common cholesterol in trade is specially purified for the process, during said process a saturated solution is produced in an organic solvent at 56-90 °C,
-then said solution is crystallized by changing the temperature, then crystals are collected on a sterile filter, dried and said crystalls are solved in another solvent and a saturated solution is produced at 56-90 °C then it is crystallized by changing the temperature,
- then collected on another sterile filter the crystals are dissolved in a third organic solvent at 56-90 °C, and are crystallized again by changing the temperature,
- crystals are collected under sterile conditions, dissolved in an organic solvent at 56-90 °C, and are stored as a supersaturated solution at 5 °C.
4. Particles produced preferably by the process according to claims any 1 to 3 characterized by that, the surface membrane of the particle of micrococcus size constitutes of cholesterol of high purity degree.
5. Particles according to claim 4 characterized by that, the structure of the membrane is retained stable in an electrolytic suspension or in a lyophilized form .
6. Particles according to any of claims 4 or 5 characterized by that, the particle encapsules part of different size, preferably organic materials of great molecules and inorganic materials jointly or separately as well.
7. Particles according to any of claims 4 to 6 characterized by that, the particle in itself, or filled with electrolytic solutions or with organic materials of great molecules is applied for active and immune-stimulation and active immunization.
8. Particles according to any of claim 4 to 7 characterized by that, they are suitable for intravenous injection into any animal (human being) whose blood can contain cholesterol among n ■ atural circumstances.
9. Particles according to any of claims 4 to 8 characterized by that, 'vacant' particles are applied for inducing production of anticholesterol antibody.
10. Particles according to any of claims 4 to 9 characterized by that, antibodies formed under influence of particles are suitable for prevention and treatment of diseases contracted due to deficiency of anticholesterol antibody or due to decline of antibody titre.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| HU0204064 | 2002-11-26 | ||
| HU0204064A HUP0204064A2 (en) | 2002-11-26 | 2002-11-26 | Process for producing bacterium sized particles derivable from cholesterol and particles produced by the process |
| PCT/HU2003/000007 WO2004047806A1 (en) | 2002-11-26 | 2003-01-31 | Process for obtaining bacterium-size particles |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP1567128A1 true EP1567128A1 (en) | 2005-08-31 |
Family
ID=89980957
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP03701627A Ceased EP1567128A1 (en) | 2002-11-26 | 2003-01-31 | Process for obtaining bacterium-size particles |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP1567128A1 (en) |
| AU (1) | AU2003202716A1 (en) |
| HU (1) | HUP0204064A2 (en) |
| WO (1) | WO2004047806A1 (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HUP0101480A2 (en) * | 2001-04-11 | 2004-01-28 | István Horváth | Process for producing particles with bacterium size from cholesterol |
-
2002
- 2002-11-26 HU HU0204064A patent/HUP0204064A2/en unknown
-
2003
- 2003-01-31 WO PCT/HU2003/000007 patent/WO2004047806A1/en not_active Application Discontinuation
- 2003-01-31 EP EP03701627A patent/EP1567128A1/en not_active Ceased
- 2003-01-31 AU AU2003202716A patent/AU2003202716A1/en not_active Abandoned
Non-Patent Citations (1)
| Title |
|---|
| See references of WO2004047806A1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| HUP0204064A2 (en) | 2005-07-28 |
| AU2003202716A1 (en) | 2004-06-18 |
| WO2004047806A1 (en) | 2004-06-10 |
| HU0204064D0 (en) | 2003-01-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0310716B1 (en) | A product, preparation and method for treating allergies | |
| NL192088C (en) | Cytotoxic product. | |
| DE69738140T2 (en) | HAPTEN-CARRIER CONJUGATES FOR USE IN DRUG THERAPY AND METHOD FOR THE PRODUCTION THEREOF | |
| AU737980B2 (en) | Novel polymerizable fatty acids, phospholipids and polymerized liposomes therefrom | |
| CN109843336A (en) | Method and composition for targeting T-cells cancer | |
| JPH04502910A (en) | Adhesive drug delivery composition | |
| EP0840796A2 (en) | Mucosal delivery of polynucleotides | |
| ES2425315A2 (en) | Combination pharmaceutical composition and methods of treating diseases or conditions associated with respiratory disease or condition | |
| LT5987B (en) | Pharmaceutical compositions | |
| JP2000510844A (en) | Method of treating type I hypersensitivity using monophosphoryl lipid A | |
| CA2067210A1 (en) | Interfacial condensation of bioactive compounds and the site-specific compounds and conjugates thereof | |
| WO1990001949A1 (en) | Gamma inulin compositions | |
| EP1567128A1 (en) | Process for obtaining bacterium-size particles | |
| CN106267198B (en) | It targets photo-thermal therapy combined immunization and treats antitumor compound formulation and the preparation method and application thereof | |
| WO2002083100A1 (en) | Liposomes consisting mainly of cholesterol | |
| Marusić-Galesić et al. | Cellular immune response to the antigen administered as an immune complex in vivo. | |
| Thakur et al. | Recent trends in targeted drug delivery | |
| JPH05501258A (en) | Methods and devices for treating medical conditions using oxidized lipoproteins | |
| JPS6221335B2 (en) | ||
| US20230381183A1 (en) | USE OF FOLIC ACID AND FOLATE MODIFICATION IN INDUCING B-CELL IMMUNE TOLERANCE AND TARGETING mIgM-POSITIVELY-EXPRESSED B-CELL LYMPHOMA | |
| RU2712212C1 (en) | Method of treating oncological diseases by drug injections | |
| WO2022071496A1 (en) | Efficacy enhancing agent for inflammatory disease treatment drug, and accumulation-promoting agent for promoting accumulation of contrast medium in inflammation site | |
| CN118027062A (en) | Rapamycin prodrug, preparation and application of nano preparation thereof | |
| Kruglov et al. | DEVELOPMENT OF THE TECHNOLOGY OF A LIPOSOMAL FORM OF EYE DROPS BASED ON A PEPTIDE COMPLEX. | |
| Nanda et al. | A novel strategy to elicit enduring anti-morphine immunity and relief from addiction by targeting Acr1 protein nano vaccine through TLR-2 to dendritic cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
| 17P | Request for examination filed |
Effective date: 20050621 |
|
| AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LI LU MC NL PT SE SI SK TR |
|
| AX | Request for extension of the european patent |
Extension state: AL LT LV MK RO |
|
| PUAJ | Public notification under rule 129 epc |
Free format text: ORIGINAL CODE: 0009425 |
|
| 32PN | Public notification |
Free format text: DECISION TO REFUSE A EUROPEAN PATENT APPLICATION (EPO FORM 2007)/27.11.2006 |
|
| STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED |
|
| 18R | Application refused |
Effective date: 20070622 |