WO2004047807A1 - Artificial blood on the basis of encapsulted haemoglobin - Google Patents
Artificial blood on the basis of encapsulted haemoglobin Download PDFInfo
- Publication number
- WO2004047807A1 WO2004047807A1 PCT/HU2003/000008 HU0300008W WO2004047807A1 WO 2004047807 A1 WO2004047807 A1 WO 2004047807A1 HU 0300008 W HU0300008 W HU 0300008W WO 2004047807 A1 WO2004047807 A1 WO 2004047807A1
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- WIPO (PCT)
- Prior art keywords
- haemoglobin
- blood
- artificial blood
- membrane
- particles
- Prior art date
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- 239000002473 artificial blood Substances 0.000 title claims abstract description 60
- 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 80
- 210000004369 blood Anatomy 0.000 claims abstract description 49
- 239000008280 blood Substances 0.000 claims abstract description 49
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 40
- 239000012528 membrane Substances 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 21
- 230000008569 process Effects 0.000 claims abstract description 21
- 241000251539 Vertebrata <Metazoa> Species 0.000 claims abstract description 12
- 239000000126 substance Substances 0.000 claims abstract description 9
- 239000002245 particle Substances 0.000 claims description 19
- 241000894007 species Species 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 11
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 10
- 239000012530 fluid Substances 0.000 claims description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 7
- 230000004087 circulation Effects 0.000 claims description 7
- 210000003743 erythrocyte Anatomy 0.000 claims description 7
- 229910052760 oxygen Inorganic materials 0.000 claims description 7
- 239000001301 oxygen Substances 0.000 claims description 7
- 210000000987 immune system Anatomy 0.000 claims description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 5
- 239000001569 carbon dioxide Substances 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 2
- 239000012466 permeate Substances 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 239000002502 liposome Substances 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 241000282414 Homo sapiens Species 0.000 description 7
- 239000013566 allergen Substances 0.000 description 6
- 230000008901 benefit Effects 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 241000124008 Mammalia Species 0.000 description 3
- 230000000172 allergic effect Effects 0.000 description 3
- 239000000823 artificial membrane Substances 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 230000002349 favourable effect Effects 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000208125 Nicotiana Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 208000010668 atopic eczema Diseases 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 235000019504 cigarettes Nutrition 0.000 description 2
- 239000003792 electrolyte Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 244000045947 parasite Species 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 206010003497 Asphyxia Diseases 0.000 description 1
- 201000011001 Ebola Hemorrhagic Fever Diseases 0.000 description 1
- 101001037191 Homo sapiens Hyaluronan synthase 1 Proteins 0.000 description 1
- 241000607149 Salmonella sp. Species 0.000 description 1
- 241000607626 Vibrio cholerae Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000002730 additional effect Effects 0.000 description 1
- 230000000879 anti-atherosclerotic effect Effects 0.000 description 1
- 230000003627 anti-cholesterol Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 238000000554 physical therapy Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009385 viral infection 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0026—Blood substitute; Oxygen transporting formulations; Plasma extender
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/41—Porphyrin- or corrin-ring-containing peptides
- A61K38/42—Haemoglobins; Myoglobins
-
- 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
Definitions
- the invention relates to a process for producing artificial blood, with haemoglobine encapsuled in a membrane as well as artificial blood produced this way, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane.
- the artificial blood is produced with using haemoglobin encapsuled in a membrane, produced from a cholesterol of high purity degree, which can be integrated in the organism of all vertebrata, but which never decomposes in the organism.
- the artificial blood produced this way is suitable for supplying blood temporarily or for long term for any mammalia - human beings and animals - of any blood group or race drained of blood.
- - Haemoglobin should be encapsuled in a membrane, which is capable of resisting permanently enzymes in the blood.
- Artificial blood offers a possibility of solving the problem of one of the most significant functions of blood, the supply of carbon dioxide and oxygen gases and supply of fluid arising due to loss of blood. Artificial blood helps the patient suffered considerable loss of blood through life danger arising due to loss of blood. The preparation due to its nature and conditions of production contains beyond doubt no infectious organisms mentioned above. With avoiding possibility of infection the patient is definitely not exposed to an additional effect endangering life.
- the invention is a process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is suitable or can be made suitable for supplying blood for all vertebrata, which is characterized by that, during the process haemoglobin is encapsuled in a pure cholesterol shell and particles are created, which can be suspended in a watery substance in a way, that haemoglobin is put between two monomolecular lipoid films of properly arranged structure made from pure cholesterol and particles are formed by splitting the plan-parallel layers in a way that haemoglobin remain suitable for supplying oxygen and carbon dioxide.
- the invention is furthermore a process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is suitable or can be made suitable for supplying blood for all vertebrata, which is characterized by that, the haemoglobin is put on a monomolecular film and the film is split in a way, that the pieces of the film create a closed surface and encapsule the haemoglobin on their surface, so the haemoglobin does not decompose during circulation.
- the invention further is artificial blood produced preferably by processes according to the invention, which is characterized by that, it involves haemoglobin encapsuled in pure cholesterol shell in the form of particles, which can be suspended in watery substance, which is suitable or can be made suitable for supplying blood for all vertebrata.
- the size of the particles remains below the size of the red-blood cells or red-blood corpuscles characteristic of the given species and said particles can permeate through the tubes of the capillaries owing to their size.
- the haemoglobin used can derive directly from a haemoglobin solution produced from blood or can be haemoglobin dried by lyophilization.
- the haemoglobin may be from the same species or deriving from other vertebrata.
- the material of the lipoid membrane encapsuling the haemoglobin is pure cholesterol.
- forming of lipoid film from pure cholesterol takes place at an optimum temperature from an optimum solution.
- forming of cholesterol membrane can take place on solid, fluid surface or in g oa' seous substance.
- a solution of pure cholesterol is produced at a proper temperature, which is suitable for making a thin monomolecular layer on a smooth surface.
- the surface of the body, the temperature, the flowing speed of the solution should be optimum.
- the smooth surface can include dents of maximum 400 nm.
- the temperature can differ from the melting temperature by maximum 10% (obviously to the higher range).
- the flowing speed of cholesterol solution can be 1 mg/sec/50cm2 +-10% on a surface of a proper angle.
- the physical impacts consistent with the viscosity can range between electromagnetic waves (270 nm) of advantageously applicable frequency and energy through sound-waves of proper wavelength and energy to inaudible frequencies.
- Fields of application of the artificial blood according to the invention a./ It ensures oxygen supply of the organism during a severe operation, while genuine blood can be preserved for intake after the operation.
- the ambulance man providing first aid can ensure oxygen supply of the organism drained of blood irrespective of blood group until the injured can get fully efficient medical treatment.
- z.l Under warring conditions it is suitable for supplying the injured soldier drained of blood with blood until he is taken to hospital. Life of the soldier can be saved by applying artificial blood.
- Canned artificial blood in lyophilized form may be the part of personal equipment. Doctors on the battlefield should only take along physiological solvent, mix it with the powdered blood and it can be immediately injectioned.
- Haemoglobin must be encapsuled into a semi-permeable lipoid film to retain its capability for supplying oxygen and carbon dioxide.
- the haemoglobm is put between two monomolecular lipoid films of properly arranged structure made from pure cholesterol and liposomes are formed by splitting the plan-parallel layers
- the other possibility is to put the haemoglobin on a monomolecular film and the film is split in a way, that the pieces of the film roll up the haemoglobin on their surface, similarly as cigarette paper covers tobacco.
- the only condition so far to meet these requirements is the novelty of our invention, using membrane made from pure cholesterol.
- Overriding immunologicai principle for the use of artificial blood is, that haemoglobin should be encapsuled in a liposome in such a way, that artificial blood should not have allergic influence. This requirement can be fulfilled so, that cholesterol existing in the organism is used in a form of high purity for forming the membrane substituting the cell membrane of the 'cells' of the artificial blood.
- the complex containing haemoglobin may assume allergen characteristics in case the artificial 'cell membrane' acting as antigen is capable of stimulating anti-bodies damaging the organism in some way. This possibility must be taken into consideration even if cholesterol does not usually appear as allergen or appears as allergen to a small extent only.
- Anti cholesterol has been known so far as anti atherosclerotic, therefore its effect is accepted as favorable. Pure cholesterol was highlighted due to the fact that there is the least chance of harming the organism.
- a liposome can be produced with applying material with the least immunologicai power which can match the features of those of the natural cells or corpuscles produced in the organism.
- the raw material of the membrane liposome of the artificial blood produced by us is compiled exclusively of pure cholesterol that can be found in every genus of vertebrata. It has no extraneous element. No allergic side effect in animal tests could be noticed.
- our achievements so far it seems realistic to achieve the set aim, i.e. our preparation will prove to be suitable for filling the function of blood as supplier of oxygen and carbon dioxide in all organisms drained of blood irrespective of blood group.
- the advantage of the process according to the invention is, that the cholesterol membrane is not decomposed by enzymes of the organism. As it is regarded as 'self by the immune system of the organism, it is not toxic in the organism of vertebrata.
- the process according to the invention makes possible production of a Liposoma Encapsulated Haemoglobin (LEH) which can live long due to the fact, that there is no enzyme in the organism that could decompose it.
- LEH particles of 200-500 nm size can easily get to every area of circulation.
- Cholesterol of high purity is suitable for encapsulating haemoglobin from any mammal organism. Due to this fact siim-le and inexpensive raw material can be ensured.
- the membrane formed from pure cholesterol solved a task, which was the objective of the future only.
- Haemoglobin does not dissolve from the particles even in case of longer storing under proper conditions.
- the artificial blood produced this way can be stored in fluid form, in liophylized state, as powder and this powder suspended suitably provides artificial blood of full value.
- the pieces of the cholesterol film produced by one of the processes of the invention create a closed surface, encapsule the haemoglobin on their surface and cover it in a similar way to a cigarette paper covering tobacco. By packaging into the film it can be achieved, that the haemoglobin should not be dissolved during circulation, because dissolved haemoglobin is secreted in the kidneys.
- LEH produced by a simple, inexpensive process can help an injured person through the danger of biological suffocation resulting from lack of blood. At the same time it restores level of fluid loss in the organism, recovering osmotic value of cells and enables haematogen organs to restore genuine composition of blood in the organism under conditions of physiological blood pressure and circulation as soon as possible.
- loss of blood is such an extent, that other constituents of blood (proteins, enzymes etc.) should be supplied without delay beside electrolytes, then there is the possibility of supplying genuine blood after artificial blood, which can be ensured under hospital conditions.
- Cholesterol did not show any variability during the phylogenesis. This phenomena has double benefits.
- the artificial blood produced is suitable not only for replacing the blood of only one species, namely that of human beings, but the same artificial blood can provide blood supply for almost all warm-blooded animals.
- the advantage of the theoretical and practical solution is, that any favorable effect recognized on an animal species can be transferred to a different species even without any modification.
- the human being, the Homo sapiens has obviously the greatest benefits from this, as the results of animal tests can be regarded identical value with observations on human beings.
Abstract
The invention relates to a process for producing artificial blood, with haemoglobine encapsuled in a membrane as well as artificial blood produced this way, which is characterized by that, it involves haemoglobin encapsuled in pure cholesterol shell in the form of particules, which can be suspended in watery substance or can be made suitable for supplying blood for all vertebrata.
Description
ARTIFICIAL BLOOD ON THE BASIS OF ENCAPSULATED HAEMOGLOBIN
The invention relates to a process for producing artificial blood, with haemoglobine encapsuled in a membrane as well as artificial blood produced this way, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane. During the process the artificial blood is produced with using haemoglobin encapsuled in a membrane, produced from a cholesterol of high purity degree, which can be integrated in the organism of all vertebrata, but which never decomposes in the organism. The artificial blood produced this way is suitable for supplying blood temporarily or for long term for any mammalia - human beings and animals - of any blood group or race drained of blood.
Demand for supplying blood had long arisen in medical science and in emergency medical attendance. The first results were achieved by restoring level of fluid content of blood in cases when considerable proportion of fluid content of blood was lost due to any physiological or pathological effects. In such cases intake of fluid ensured restoring the patient from severe cases and created favorable conditions for healing. In case of young children or elderly people it is very important to take this aspect into consideration, because loss of fluid could be fatal for the endangered organism. Often giving water can no longer help. In such cases patients should be supplied with water infusion injected directly to circulation. Prevention of drying out could be life saving in case of both young children and adults belonging to any age group suffering from infection caused by Salmonella sp., Vibrio cholera, certain viruses etc.
During loss of blood three essential basic materials leave the organism:
- Water, the fluid component of blood together with electrolytes solved in water.
- Proteins together with enzymes and antibodies.
- Cell elements, among them red blood cells accounting for supplying gases.
Possibility of blood transfusion has been made use of in almost every medical treatment as well as on battlefields since Landsteiner elaborated it in the knowledge of blood groups. Today, when pharmaceutical industry producing, and doctors using blood preparations must pay particular attention to the prevention of today still incurable virus-infections, applying of blood preparations or blood- supplementing materials offering provisional solutions are of particular significance.
In certain publications the following future tasks of producing artificial blood were designated:
- Haemoglobin should be encapsuled in a membrane, which is capable of resisting permanently enzymes in the blood.
- At the same time it should be a material acceptable for every organism, it should be regarded as 'self by the organism.
In the state of art earlier processes are known, during which artificial membranes were produced trying to imitate chemical constituents of membranes in the organism. 19-22 % cholesterol could be found in the artificial membranes involving haemoglobin. Membranes built up mainly by using mixed lipoids, mainly phospholipids could not resist decomposing effect of enzymes. Combinations of lipoids promoted forming of artificial membranes, whereas membranes of pure cholesterol were difficult to form.
Artificial blood offers a possibility of solving the problem of one of the most significant functions of blood, the supply of carbon dioxide and oxygen gases and supply of fluid arising due to loss of blood. Artificial blood helps the patient suffered considerable loss of blood through life danger arising due to loss of blood. The preparation due to its nature and conditions of production contains beyond doubt no infectious organisms mentioned above. With avoiding possibility of infection the patient is definitely not exposed to an additional effect endangering life.
In the state of art the importance and significance of producing artificial blood have been recognized by many and several solutions were found to produce artificial blood with haemoglobin encapsuled in a liposome membrane (Djordjevich et al. PCT/US82/00899), and it can be clearly seen from works summerizing information in connection with artificial blood (Andreas M. Spiring et al: Drug of the future 1993. 18. (3) 249-253) that in forming the membrane of liposome several materials, first of all phospholipid are used.
The solutions available suffer from the limitation, that the membrane of the 'cells' was built of materials, which could be decomposed by enzymes and several of its lipoids might have an allergic effect.
When elaborating the solution according to the invention we aimed to work out a process as well as a blood supplementing suspension, in which haemoglobin is encapsuled in a membrane, which can not be decomposed by enzymes in the blood.
We realized, when we worked out the solution according to the invention, that a membrane made. from pure cholesterol meets these requirements fully, in case we use a single material for producing the membrane contrary to the processes in the state of art, and this material is cholesterol. So far cholesterol has not been applied for producing membranes in itself in pure state. In water solution, e.g. in a haemoglobin solution cholesterol crystallizes and is not able to produce a membrane which prevents it from being used for membrane production.
The invention is a process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is
suitable or can be made suitable for supplying blood for all vertebrata, which is characterized by that, during the process haemoglobin is encapsuled in a pure cholesterol shell and particles are created, which can be suspended in a watery substance in a way, that haemoglobin is put between two monomolecular lipoid films of properly arranged structure made from pure cholesterol and particles are formed by splitting the plan-parallel layers in a way that haemoglobin remain suitable for supplying oxygen and carbon dioxide.
The invention is furthermore a process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is suitable or can be made suitable for supplying blood for all vertebrata, which is characterized by that, the haemoglobin is put on a monomolecular film and the film is split in a way, that the pieces of the film create a closed surface and encapsule the haemoglobin on their surface, so the haemoglobin does not decompose during circulation.
The invention further is artificial blood produced preferably by processes according to the invention, which is characterized by that, it involves haemoglobin encapsuled in pure cholesterol shell in the form of particles, which can be suspended in watery substance, which is suitable or can be made suitable for supplying blood for all vertebrata.
In one of the preferred realizations of the artificial blood according to the invention, the size of the particles remains below the size of the red-blood cells or red-blood corpuscles characteristic of the given species and said particles can permeate through the tubes of the capillaries owing to their size.
In another preferred realization of the artificial blood according to the invention, the haemoglobin used can derive directly from a haemoglobin solution produced from blood or can be haemoglobin dried by lyophilization.
In a further preferred realization of the artificial blood according to the invention, from the point of view of application of artificial blood the haemoglobin may be from the same species or deriving from other vertebrata.
In another preferred realization of the artificial blood according to the invention, during forming of the particles the material of the lipoid membrane encapsuling the haemoglobin is pure cholesterol.
In another preferred realization of the artificial blood according to the invention, forming of lipoid film from pure cholesterol takes place at an optimum temperature from an optimum solution.
In another preferred realization of the artificial blood according to the invention, forming of cholesterol membrane can take place on solid, fluid surface or in g oa' seous substance.
In another preferred realization of the artificial blood according to the invention, during forming of particles splitting of the membrane into particle size takes place under the influence of variable physical energy.
The solution according to the invention is set forth by the following examples:
Example 1
Production of artificial blood
A solution of pure cholesterol is produced at a proper temperature, which is suitable for making a thin monomolecular layer on a smooth surface. The surface of the body, the temperature, the flowing speed of the solution should be optimum. The smooth surface can include dents of maximum 400 nm. The temperature can differ from the melting temperature by maximum 10% (obviously to the higher range). The flowing speed of cholesterol solution can be 1 mg/sec/50cm2 +-10% on a surface of a proper angle. The physical impacts consistent with the viscosity can range between electromagnetic waves (270 nm) of advantageously applicable frequency and energy through sound-waves of proper wavelength and energy to inaudible frequencies.
Determining the suitable frequency and wavelength takes place with choosing the values with adjustable equipment complying best with the temperature and angular offset. We bring haemoglobin molecules in connection with the cholesterol solution under ideal conditions for producing liposome. The physical force arising at the moment of encounter splits the cholesterol layer in the given size and it encapsules the haemoglobin. Liposomes produced this way are put into a liquid of around pH 7 value and are separated from haemoglobin molecules soluble or remained outside the lipoid shell. The haemoglobin solution is regained and the liposomes containing haemoglobin are collected for producing artificial blood.
Example 2
Fields of application of the artificial blood according to the invention: a./ It ensures oxygen supply of the organism during a severe operation, while genuine blood can be preserved for intake after the operation. b./ In case of an accident the ambulance man providing first aid can ensure oxygen supply of the organism drained of blood irrespective of blood group until the injured can get fully efficient medical treatment. z.l Under warring conditions it is suitable for supplying the injured soldier drained of blood with blood until he is taken to hospital. Life of the soldier can be saved by applying artificial blood. Canned artificial blood in lyophilized form may be the
part of personal equipment. Doctors on the battlefield should only take along physiological solvent, mix it with the powdered blood and it can be immediately injectioned. d./ In case of special therapy it is suitable for replacing blood during a relatively short physical therapy, (chemotherapy, radiotherapy) e/ parasites of red blood cells (malaria, Babezia etc.) can not attack artificial blood produced according to the invention with haemoglobin encapsuled in a liposome
(HEL). f / gas supply can be increased in an organism demanding high oxygen consumption.
Example 3
Physical principle of production of artificial blood according to the invention:
Haemoglobin must be encapsuled into a semi-permeable lipoid film to retain its capability for supplying oxygen and carbon dioxide. During the process the haemoglobm is put between two monomolecular lipoid films of properly arranged structure made from pure cholesterol and liposomes are formed by splitting the plan-parallel layers The other possibility is to put the haemoglobin on a monomolecular film and the film is split in a way, that the pieces of the film roll up the haemoglobin on their surface, similarly as cigarette paper covers tobacco. The only condition so far to meet these requirements is the novelty of our invention, using membrane made from pure cholesterol. By packaging into the film it can be achieved, that the haemoglobin should not be dissolved during circulation, because dissolved haemoglobin is secreted in the kidneys.
The main characteristics of the artificial blood made by the process according to the invention are as follows:
- Haemoglobin of any animal species can be used
- Artificial blood can be applied in any animal species
- Pure cholesterol does not decompose in the organism, therefore circulation is ensured for a long term.
Cholesterol shell does not act as allergen
- Particles can be well lyophilized
- Blood parasites endangering red blood cells can not attack the artificial blood.
Immunologicai conditions of producing artificial blood :
Overriding immunologicai principle for the use of artificial blood is, that haemoglobin should be encapsuled in a liposome in such a way, that artificial blood should not have allergic influence. This requirement can be fulfilled so, that cholesterol existing in the organism is used in a form of high purity for forming the membrane substituting the cell membrane of the 'cells' of the artificial blood.
Difficulties entitled with the task:
The complex containing haemoglobin may assume allergen characteristics in case the artificial 'cell membrane' acting as antigen is capable of stimulating anti-bodies damaging the organism in some way. This possibility must be taken into consideration even if cholesterol does not usually appear as allergen or appears as allergen to a small extent only. Nowadays it is not infrequent that materials earlier not considered allergen by allergologists in fact act as allergens. Anti cholesterol has been known so far as anti atherosclerotic, therefore its effect is accepted as favorable. Pure cholesterol was highlighted due to the fact that there is the least chance of harming the organism.
Possibility of overcoming side-effects
Our observations so far support our earlier working hypothesis, that a liposome can be produced with applying material with the least immunologicai power which can match the features of those of the natural cells or corpuscles produced in the organism. The raw material of the membrane liposome of the artificial blood produced by us is compiled exclusively of pure cholesterol that can be found in every genus of vertebrata. It has no extraneous element. No allergic side effect in animal tests could be noticed. On basis of our achievements so far it seems realistic to achieve the set aim, i.e. our preparation will prove to be suitable for filling the function of blood as supplier of oxygen and carbon dioxide in all organisms drained of blood irrespective of blood group.
As mentioned in the introduction, it is first of all suitable for meeting the demands of casualties and operations lasting for a relatively short time, but in a certain circle it ensures solution for medical treatment as well. On basis of the results of the animal tests carried out so far and keeping in mind the theoretical considerations already mentioned we find it suitable for use for human beings as well.
The advantage of the process according to the invention is, that the cholesterol membrane is not decomposed by enzymes of the organism. As it is regarded as 'self by the immune system of the organism, it is not toxic in the organism of vertebrata. The process according to the invention makes possible production of a Liposoma Encapsulated Haemoglobin (LEH) which can live long due to the fact, that there is no enzyme in the organism that could decompose it. The LEH particles of 200-500 nm size can easily get to every area of circulation. Cholesterol of high purity is suitable for encapsulating haemoglobin from any mammal organism. Due to this fact siim-le and inexpensive raw material can be ensured.
The membrane formed from pure cholesterol solved a task, which was the objective of the future only. Haemoglobin does not dissolve from the particles even in case of longer storing under proper conditions. The artificial blood produced this way can be stored in fluid form, in liophylized state, as powder and this powder suspended suitably provides artificial blood of full value.
The pieces of the cholesterol film produced by one of the processes of the invention create a closed surface, encapsule the haemoglobin on their surface and cover it in a similar way to a cigarette paper covering tobacco. By packaging into the film it can be achieved, that the haemoglobin should not be dissolved during circulation, because dissolved haemoglobin is secreted in the kidneys.
LEH produced by a simple, inexpensive process can help an injured person through the danger of biological suffocation resulting from lack of blood. At the same time it restores level of fluid loss in the organism, recovering osmotic value of cells and enables haematogen organs to restore genuine composition of blood in the organism under conditions of physiological blood pressure and circulation as soon as possible. In case loss of blood is such an extent, that other constituents of blood (proteins, enzymes etc.) should be supplied without delay beside electrolytes, then there is the possibility of supplying genuine blood after artificial blood, which can be ensured under hospital conditions.
Solving the problem of artificial blood meets the requirements of an ancient demand, which seemed unreal many times, and which is one of the most significant achievements in sorting out problems arising due to loss of blood since the application of blood transfusion elaborated by Landsteiner armed with the knowledge of blood groups. The mutual possibility of application of haemoglobins of different species can be solved by applying artificial blood.
With the help of the process elaborated by us 'blood corpuscles" are given to the users, which creates possibility of making use of foreign species beside using it within the species. The possibility of using artificial blood mutually is given by the cholesterol, which can be found in every species with identical chemical constituents. As cholesterol acts in every mammal as 'self, it can not be decomposed in any species, haemoglobin encapsuled in cholesterol membrane can be used in every species providing artificial blood for the needy.
Further research can also benefit from this invention as well as users. Making use of corpuscles produced with using haemoglobm strange for species the needy can get access to artificial blood, which can be made in unlimited quantity. This can be done in a way, that presence and transfer of microorganisms endangering human organism (HIN, Ebola etc.) can be excluded with great safety.
Cholesterol did not show any variability during the phylogenesis. This phenomena has double benefits. The artificial blood produced is suitable not only for replacing the blood of only one species, namely that of human beings, but the same artificial blood can provide blood supply for almost all warm-blooded animals. The advantage of the theoretical and practical solution is, that any favorable effect recognized on an animal species can be transferred to a different species even
without any modification. The human being, the Homo sapiens has obviously the greatest benefits from this, as the results of animal tests can be regarded identical value with observations on human beings.
Claims
Process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is suitable or can be made suitable for supplying blood for all vertebrata, characterized by that, during the process haemoglobin is encapsuled in a pure cholesterol shell and particles are created, which can be suspended in a watery substance in a way, that haemoglobin is put between two monomolecular lipoid films of properly arranged structure made from pure cholesterol and particles are formed by splitting the plan-parallel layers in a way that haemoglobin remain suitable for supplying oxygen and carbon dioxide.
2. Process for producing artificial blood, which does not decompose in the organism, - but which is regarded as 'self by the immune system of the organism- from haemoglobine encapsuled in a membrane, which is suitable or can be made suitable for supplying blood for all vertebrata, characterized by that, the haemoglobin is put on a monomolecular film and the film is split in a way, that the pieces of the film create a closed surface and encapsule the haemoglobin on their surface, so the haemoglobin does not decompose during circulation.
3. Artificial blood produced preferably by processes according to claim 1 or 2, characterized by that, it involves haemoglobin encapsuled in pure cholesterol shell in the form of particles, which can be suspended in watery substance, which is suitable or can be made suitable for supplying blood for all vertebrata.
4. Artificial blood according to claim 3 characterized by that, the size of the particles remains below the size of the red-blood cells or red-blood corpuscles characteristic of the given species and said particles can permeate through the tubes of the capillaries owing to their size.
5. Artificial blood according to any of claims 1 to 4 characterized by that, the haemoglobin used can derive directly from a haemoglobin solution produced from blood or can be haemoglobin dried by lyophilization.
6. Artificial blood according to any of claims 1 to 5 characterized by that from the point of view of application of artificial blood the haemoglobin may be from the same species or deriving from other vertebrata.
7. A-rtificial blood according to any of claims 1 to 6 characterized by that during forming of the particles the material of the lipoid membrane encapsuling the haemoglobin is pure cholesterol.
8. Artificial blood according to claim 7 characterized by that, forming of lipoid film from pure cholesterol takes place at an optimum temperature from an optimum solution.
9. Artificial blood according to claim 6 characterized by that, forming of cholesterol membrane can take place on solid, fluid surface or in gaseous substance.
10. Artificial blood according to any of claims 3 to 9 characterized by that, during forming of particles splitting of the membrane into particle size takes place under the influence of variable physical energy.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2003202717A AU2003202717A1 (en) | 2002-11-26 | 2003-01-31 | Artificial blood on the basis of encapsulted haemoglobin |
| EP03701628A EP1567129A1 (en) | 2002-11-26 | 2003-01-31 | Artificial blood on the basis of encapsulated haemoglobin |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| HU0204065A HU226519B1 (en) | 2002-11-26 | 2002-11-26 | Artificial blood made of hemoglobin encapsulated in membrane |
| HUP0204065 | 2002-11-26 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2004047807A1 true WO2004047807A1 (en) | 2004-06-10 |
Family
ID=89980958
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/HU2003/000008 WO2004047807A1 (en) | 2002-11-26 | 2003-01-31 | Artificial blood on the basis of encapsulted haemoglobin |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP1567129A1 (en) |
| AU (1) | AU2003202717A1 (en) |
| HU (1) | HU226519B1 (en) |
| WO (1) | WO2004047807A1 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4133874A (en) * | 1976-06-10 | 1979-01-09 | The University Of Illinois Foundation | Lipid encapsulated hemoglobin cells |
| US4891208A (en) * | 1985-04-10 | 1990-01-02 | The Liposome Company, Inc. | Steroidal liposomes |
| WO2002087540A1 (en) * | 2001-04-27 | 2002-11-07 | Horvath Istvan | Artificial blood made of hemoglobin encapsulated in liposomes |
-
2002
- 2002-11-26 HU HU0204065A patent/HU226519B1/en not_active IP Right Cessation
-
2003
- 2003-01-31 EP EP03701628A patent/EP1567129A1/en not_active Withdrawn
- 2003-01-31 AU AU2003202717A patent/AU2003202717A1/en not_active Abandoned
- 2003-01-31 WO PCT/HU2003/000008 patent/WO2004047807A1/en not_active Application Discontinuation
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4133874A (en) * | 1976-06-10 | 1979-01-09 | The University Of Illinois Foundation | Lipid encapsulated hemoglobin cells |
| US4891208A (en) * | 1985-04-10 | 1990-01-02 | The Liposome Company, Inc. | Steroidal liposomes |
| WO2002087540A1 (en) * | 2001-04-27 | 2002-11-07 | Horvath Istvan | Artificial blood made of hemoglobin encapsulated in liposomes |
Non-Patent Citations (4)
| Title |
|---|
| BIOCHIMICA ET BIOPHYSICA ACTA, vol. 692, no. 2, 1982, pages 227 - 232, ISSN: 0006-3002 * |
| DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1982, BROCKERHOFF H ET AL: "PREPARATION AND STRUCTURAL STUDIES OF CHOLESTEROL BI LAYERS", XP002245508, Database accession no. PREV198376015956 * |
| GOINS B ET AL: "LIPOSOME-ENCAPSULATED HEMOGLOBIN: HISTORICAL DEVELOPMENT OF A BLOOD SUBSTITUTE", BIOTECHNOLOGY, BUTTERWORTHS, LONDON, GB, vol. 19, 1991, pages 117 - 125, XP001097836, ISSN: 0740-7378 * |
| SPIRIG A M ET AL: "LIPOSOME ENCAPSULATED HEMOGLOBIN: A POTENTIAL ARTIFICIAL BLOOD SUBSTITUTE", DRUGS OF THE FUTURE, BARCELONA, ES, vol. 18, no. 3, 1993, pages 249 - 253, XP001097845, ISSN: 0377-8282 * |
Also Published As
| Publication number | Publication date |
|---|---|
| EP1567129A1 (en) | 2005-08-31 |
| AU2003202717A1 (en) | 2004-06-18 |
| HU0204065D0 (en) | 2003-01-28 |
| HUP0204065A2 (en) | 2004-08-30 |
| HU226519B1 (en) | 2009-03-30 |
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