DIKMEN NURTEN (TR)
GUZEL BILGEHAN (TR)
ULUSAL FATMA (TR)
YILGOR HURI PINAR (TR)
| US20140295410A1 | 2014-10-02 | |||
| CN105712938A | 2016-06-29 |
| CLAIMS 1) A method of producing nanotechnological biomaterials alternative to blood with the ability to bind haemoglobin, enzymes and drugs from magnetic nanoparticles, characterized by the steps below; - carboxylic acid (8) is added to (7) magnetic nanoparticles, which are suspended by an ultrasonic bath in pure water, - mixing with an ultrasonic stirrer (9), - reduction addition to the obtained carboxylic acid-coated magnetic nanoparticles (11), - Suspension of the obtained surface alcohol-coated nanoparticles by mixing with ultrasonic bath in acetone medium (13), - the alcohol surfaces are characterized by the outlet stages of the aldehyde-surfaced nanoparticles (15), which are the second basic process step, by the addition of an oxidant (14). 2) A method of producing nanotechnological biomaterials alternative to blood with the ability to bind haemoglobin, enzymes and drugs from magnetic nanoparticles, characterized by the steps below; - Adding alcohol derivative coating agent (21 ), - mixing with an ultrasonic stirrer (9), - Performing magnetic decantation (3), - Performing washing with pure water (4), - Performing washing with acetone (22), - obtaining magnetic nanoparticle (24) coated with alcohol derived agent by drying in the oven (23), - Mixing synthesized particles with ultrasonic bath in acetone (13) to form aldehyde-coated magnetic nanoparticle (15), - Adding an oxidant (14), - mixing with an ultrasonic stirrer (9), - Washing with pure water (4), - drying in the vacuum desiccator (5). 3) A method of producing nanotechnological biomaterials alternative to blood with the ability to bind haemoglobin, enzymes and drugs from magnetic nanoparticles, characterized by the steps below; - Suspension by an ultrasonic bath in pure water (7), - addition of aldehyde (25), - mixing with an ultrasonic stirrer (9), - Performing magnetic decantation (3), - Washing with pure water (22), - Drying in the vacuum desiccator (5). 4) A method of producing nanotechnological biomaterials alternative to blood with the ability to bind haemoglobin, enzymes and drugs from magnetic nanoparticles, characterized by the steps below; - Mixing in an ultrasonic bath in acetone (13), - Adding silane derivative coating agent (26), - Suspension by an ultrasonic bath in pH 7.4 (16), - addition of aldehyde (25), - mixing with an ultrasonic stirrer (9), - Performing magnetic decantation (3), - Washing with pure water (4), - Drying in the oven (23). 5) The process step of adding the reducer of Claim 1 , characterized in that NaBhU (Sodium Boron Hydride), UAIH4 (lithium aluminium hydride), CH4N2O2S (Thiourea), Na2S204 (hydrosulphide) solutions are used as reducers. 6) The addition of the oxidants of claims 1 and 2 characterized in that the solutions of pyridinium chlorochromate, potassium dichromate, sodium dichromate, ammonium dichromate, sodium permanganate, potassium permanganate and ammonium permanganate are used as oxidants. 7) The phase of adding carboxylic acid of Claim 1 characterized in that; comprising tartaric acid, azelaic acid, malic acid, succinic acid, butyric acid, adipic acid, oxaloacetic acid, sebacic acid, aspartic acid, glyoxal, glutaraldehyde, succinynaldehyde, malondialdehyde as dialdehyde, alcohol (Pentan-1,5-diol, triethanolamine, propane-1, 2, 3-triol, ethane-1 ,1 ,2-triol, 3- (hydroxymethyl)pentane-l ,2,5-triol) and silane ((3-Aminopropyl) triethoxysilane, (3-Aminopropyl) trimethoxysilane, N-(2-Aminoethyl)-3- aminopropropyltriethoxysilane, N, N-Bis(2-Hydroxypropyltriethyl)-3- aminopropyltriethylethoxysilane as carboxylic acid. |
AND DRUG
DESCRIPTION
TECHNICAL FIELD
The present invention is related to a method of producing nano-technological magnetic bio-materials, which are in vitro produced in the laboratory, capable of being used in place of blood, carries oxygen, and has an active enzymatic function in drug targeting.
PRIOR ART
Regardless of the blood group, the search for a suitable substance that can be safely transported anywhere and at any time and can be stored on the shelf for a long period of time has taken years. By rule, the transfusion of any substance other than the autologous blood into circulation is the transfusion of a substance that replaces blood. Therefore, the history of blood transplant can be viewed as a history of blood substitute material. Milk, casein derivatives, starch, saline and ringer solution are tried before the first successful blood transfer from person to person. Since the Second World War, the search for an alternative substance to replace blood has increased to cope with war-like situations and major civil disasters. The function of each blood component and its replacement are well known, except for its ability to carry oxygen. Over time, there has been a significant improvement in the development of these blood-substitute substances.
Eliminating unwanted side effects, especially transfusion-inflammatory diseases (HIV and Hepatitis) and leukocyte-broker alio sensitivity, is a key objective of modern transfusion medicine. The high cost of blood collection and conservation, the global lack of reserves and other problems are driving forces leading to the development of blood substitute substances. The research areas in this area are headed by haemoglobin-based oxygen carriers.
Although they do not exactly replace perfectly-designed red blood cells, these “oxygen carrier solutions” have many potential areas of use, both clinically and not clinically. These substances can reach the tissues more easily than normal red blood cells and directly transmit oxygen to the tissue. These substances may also have side effects. Extensive clinical studies are conducted on their reliability and benefits. A better understanding of the impact patterns of these products will help to identify their practices and usefulness. However, once clinical studies have been approved, they will be applicable to patients.
PURPOSE OF THE INVENTION
The purpose of the present invention is to develop of a magnetic artificial blood biomaterial, an alternative to natural blood, to be used as a preoperative and operative. It is also the process of making modifications to the surface of synthesized iron oxide solid support, in order to develop medical and medical care equipment and to target drugs.
Development of a solution-based material that can transport oxygen into a nanotechnology-based tissue with life-saving potential, regardless of blood groups, is ensured in emergencies where natural blood donor is not available. This allows an effective solution to be developed based on the frequency of the presence of natural blood used today, so that it can be immediately applied to every person with bleeding, regardless of their blood antigen. In vivo applications, reducing cellular damage, reducing carbon dioxide accumulated in the body, the stability of all types of enzymes, the presence of magnetic drug targeting floors in the target area also indicates that this solution can exceed many problems.
LIST OF THE FIGURES
Figure 1 : General process flow diagram of a natural blood alternative solution capable of carrying oxygen, targeting drugs, and removing carbon dioxide produced by iron oxide magnetite (magnetic nanoparticles) and pure haemoglobin molecules
Description of the references in the figures
1. Dissolution of FeCh and FeS04 salts
2. Sodium hydroxide precipitation in the nitrogen atmosphere
3. Magnetic decantation
4. Washing with distilled water
5. Drying in a vacuum desiccator
6. Magnetic nanoparticle 7. Suspension by an ultrasonic bath in pure water
8. Carboxylic acid
9. Mixing with ultrasonic bath
10. Carboxylic acid-coated magnetic nanoparticle 11. Adding a reducing agent
12. Alcohol coated magnetic nanoparticle (on surface)
13. Mixing with ultrasonic bath in acetone
14. Adding an oxidant
15. Aldehyde coated magnetic nanoparticle (on surface) 16. Suspension with an ultrasonic bath in pH 7.4 buffer solution
17. Addition of ultra-pure haemoglobin or drug or enzyme
18. Blending with a rotator
19. Washing with pH 7,4 buffer solution
20. Preparation of haemoglobin or enzyme complex or drug-immobilized magnetic nanoparticle
21. Adding of alcohol derivative coating agent
22. Washing with acetone
23. Drying in the oven
24. Magnetic nanoparticle coated with alcohol derivative 25. Addition of aldehyde
26. Adding silane derivative coating agent
27. Magnetic nanoparticle coated with silane derivative
A. An entire process of producing a blood-alternative, nano-technological biomaterial capable of bonding haemoglobin, enzyme and drugs
DETAILED DESCRIPTION OF THE INVENTION
The process of processing in the present invention is basically three steps:
- Preparation of magnetic nanoparticle (6)
- Preparation of aldehyde coated magnetic nanoparticle (on surface) (15), - Preparation of haemoglobin or enzyme complex or drug-immobilized magnetic nanoparticle (20)
These basic steps are accompanied by several intermediate steps. All steps are shown in the flow diagram in Figure 1. When all process steps have been examined (A), magnetic nanoparticles have been synthesized by precipitating (2) with NaOH (sodium hydroxide), KOH (potassium hydroxide), Nhh (ammonia) in the nitrogen atmosphere with the sequential steps of magnetic decantation (3), washing with pure water (4) and washing in vacuum desiccator (5) with reference to the dissolution (1) of the salts in the form of iron II and III (6). Four different methods have been used to transform the synthesized material into aldehyde-coated nanoparticles (15), the second basic step. In the first method, the magnetic nanoparticles (7), which are suspended by an ultrasonic bath in pure water, are mixed with carboxylic acid (8) and by an ultrasonic bath (9). Tartaric acid, azelaic acid, malic acid, succinic acid, butyric acid, adipic acid, oxaloacetic acid, sebacicacid, aspartic acid, glyoxal, glutaraldehyde, succinynaldehyde, malondialdehyde as dialdehyde, alcohol (Pentan-1 ,5-diol, triethanolamine, propane-1, 2, 3-triol, ethane- 1, 1 ,2-triol, 3-(hydroxymethyl)pentane-1 ,2,5-triol) and silane ((3-Aminopropyl) triethoxysilane, (3-Aminopropyl) trimethoxysilane, N-(2-Aminoethyl)-3- aminopropropyltriethoxysilane, N, N-Bis(2-Hydroxypropyltriethyl)-3- aminopropyltriethylethoxysilane have been used as carboxylic acid. Carboxylic acid is preferred because it has the ability to bind biological molecules with direct intermolecular interactions and is suitable for modification. The reason for selecting the silanes is that it has the ability to form an imine bond with the groups with aldehyde group due to the presence of amine group at the ends and it is easy to modify with a simple molecule such as glutaraldehyde. Alcohols are also suitable for weak intermolecular interactions and easy to convert to a functional group such as aldehyde by a simple method. The reason for the use of dialdehydes is preferred due to the fact that they fall down to a single step for binding of magnetic nanoparticles and biological molecules. In this way, the carboxylic acid-coated magnetic nanoparticle has been obtained (10) and the surface alcohol coated nanoparticles have been obtained by reducing with the addition of reducer (11 ) (12). NaBFU (Sodium Boron Hydride), UAIH4 (lithium aluminium hydride), Thiourea (CH4N2O2S), hydrosulphite (Na2S204) solutions have been used as reducers. The reason for the preference of these reducers is that they are strong and can reduce carboxylic acids up to the alcohol step. In this way, since the functional group will be alcohol, it can be used directly as a support material and it will be easy to convert to the aldehyde group. In addition, the preferred reducers are available in terms of toxic effects. By mixing these obtained particles with ultrasonic bath (13) in acetone medium and providing oxidation with the addition of oxidants (14), alcohol surfaces provided the outlet of aldehyde surface nanoparticles (15), the second basic process step. Pyridinium chlorochromate, potassium dichromate, sodium dichromate, ammonium dichromate, sodium permanganate, potassium permanganate and ammonium permanganate solutions have been used as oxidants. The preferred reason for the oxidants used is to ensure that the oxidation of alcohols takes place in a single step and stays on the aldehyde step. In this way, the aldehyde functional group will be obtained.
The second method of making aldehyde coated magnetic nanoparticle (15) has been the preparation of magnetic nanoparticle (6) followed by the addition of alcohol derivative coating agent (21), and the magnetic nanoparticle (24) coated with alcohol derivative agent has been obtained by mixing with ultrasonic bath (9), magnetic decantation (3) and washing with pure water (4) intermediate steps followed respectively by acetone washing (22) and oven drying (23). For the transformation of these synthesized particles into aldehyde-coated magnetic nanoparticles (15), mixing with ultrasonic bath (13), adding oxidants (14), mixing with ultrasonic stirrer (9), washing with pure water (4) and drying steps in vacuum desiccator (5) have been applied in acetone as in the previous method. Pyridinium chlorochromate, potassium dichromate, sodium dichromate, ammonium dichromate, sodium permanganate, potassium permanganate and ammonium permanganate solutions have been used as oxidants. The third method for making aldehyde coated magnetic nanoparticle (on surface) (15) has been the preparation of magnetic nanoparticles (6) and their suspension with ultrasonic bath (7) in pure water, followed by intermediate steps (9), magnetic decantation (3), washing with acetone (22) and drying in vacuum desiccator (5) with mixing with the ultrasonic stirrer applied respectively, the surface of these particles could be converted into aldehyde-coated magnetic nanoparticle (on surface) (15).
The fourth and final method of making aldehyde coated magnetic nanoparticle (15) has been to prepare magnetic nanoparticle (6) and then mix it in acetone in an ultrasonic bath (13) and add silane derivative coating agent (26). Following the steps of suspension with ultrasonic bath (16), addition of aldehyde (25), mixing with ultrasonic stirrer (9), magnetic decantation (3), washing with pure water (4), and drying in the oven (23), respectively, the aldehyde coated magnetic nanoparticle (15) and the silane derivative agent could be transformed into coated magnetic nanoparticle (27). The aldehyde coated magnetic nanoparticles (15) synthesized over four different methods have been washed (19) with the steps of suspension (16) with ultrasonic bath at pH 7.4, addition of ultrapure haemoglobin, appropriate enzyme and drug (17), mixing with rotator stirrer (18) and magnetic decantation (3) with pH 7.4 buffer. In this way, solutions could be obtained with the preparation of haemoglobin, enzyme complex and drug immobilized magnetic nanoparticles (20) on the surface of our study, which has surface modifications with oxygen, enzyme and drug transport properties (A). Manner of application of the present invention to industry
Surface modifications serving the above-mentioned purposes, haemoglobin, enzyme and drug complex solutions bound to magnetic nanoparticles can be produced, used and applied in industry.
With the determination of the toxic effect of natural haemoglobin on human use, modified haemoglobin development studies have been focused and these studies have been increasing day by day. In artificial blood transfusion studies in rat animals, haemoglobin solutions prepared by haemolysis of red blood cells have been tried; it has been determined that this solution could carry oxygen to tissues but showed toxic effects in the liver and caused high blood pressure. The absence of antigen properties of this material that we synthesize does not cause immune system problems. Therefore, it can be used effectively in case of industrialization.
In addition, phase 1 blood transfusion studies with haemoglobin solution have shown toxic effects on the kidneys and vasoactive structure. Co-administration with antioxidant enzyme systems added to the material we synthesize has provided a potential to reduce cytotoxic damage, which will be one of the reasons why it is preferred in industrial applications.
Von Stark (1898) first used the haemoglobin solution to treat an anaemia patient. Subsequent research has made changes to haemoglobin to produce purified haemoglobin. Recently, haemoglobin-based oxygen carrier solutions represent the class of substances that are alternatives to natural blood. The therapeutic purpose of these compounds is to prevent or reduce blood transfusions in cases of medical acute haemoglobin insufficiency and in different surgical operations. Their main advantages include large quantities, long storage, rapid application (without typing and cross matching) and sterilization by pasteurization. In this type of haemoglobin-based artificial blood studies, modifications have been mostly based on intra-molecular, inter-molecular or recombinant haemoglobin molecules. It has also been directed to respirocytes, peril uorocarbon, stem cell studies as well as haemoglobin-based solutions in artificial blood studies. Furthermore, the ability to bind drugs to magnetic nanoparticles plays an active role in targeting drugs to cancerous tissue. Thanks to its enzyme binding feature, maintaining its effectiveness even at high temperatures without denaturing will certainly create a basis for many medical and therapeutic equipment such as eliza, imaging, targeting in medical and medical fields. In the present invention, surface modifications of iron oxide nanoparticles synthesized using various chemical materials have been performed and ultrapure haemoglobin molecules, drugs and enzymes have been immobilized to the surface of nanoparticles undergoing chemical surface modification. Immobilized haemoglobin molecules, drug and enzyme systems thus remain stable and maintain their functional properties. The shelf life of the nanoparticle-based haemoglobin solution thus stabilized is also long.