JPH05500963A - Novel oxidized fat protein and its preparation method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Novel oxidized fat protein and its preparation method Background of the invention Cross-reference of related applications This application is filed in U.S. Patent Application No. 07/418.382, filed October 6, 1989. This is a partial continuation application of No.

Declaration of federal funding Funding for the research described herein was provided by the federal government under authorization from the Health and Human Services Administration. offered. Accordingly, the federal government has certain rights in this invention.

Conventional technology The use of oxidized fatty proteins to treat diseases is described in detail in the above-mentioned patent applications. It is listed here as a reference. Also listed as a reference However, another teaching is ``Using Nuclear Magnetic Resonance'' by Eric I. Couchy and Fossel. "Means for testing cancer" (U.S. patent filed on March 27, 1990) No. 4.912.050) and 'Cancer Detection Means Using Nuclear Magnetic Resonance' (No. 19 United States Patent Army No. 4,918.021 filed on April 17, 1990) There is.

field of invention The present invention relates to novel oxidized fat protein compositions, and in particular to novel oxidized fat protein compositions. The present invention relates to a method for preparing a product and a device for preparing the same.

Most organic compounds, including most cellular components, are thermodynamically It is in an unstable state. The direct reaction of the normal triplet state of oxygen with a methylene group Because there are both energy barriers and electron spin barriers for the reaction, , the reaction is generally very gradual. However, in cells Some of the existing substances react relatively quickly with oxygen.

Among these substances are unsaturated fatty acids.

One of the known properties of this unsaturated fatty acid, especially polyenoyl monolytic acid, is that oxidation , especially susceptibility to peroxidation and autoxidation. What is autoxidation? One of the processes is a radical chain reaction involving molecular oxygen as a reactant. Ru. The term ``peroxide'' refers to products from peroxides and their decomposition products. be. Generally, the term oxide contains one oxygen molecule, whereas peroxide Contains two oxygen molecules. According to the present invention, another form of oxidation, photocatalytic oxidation, provided. This form of oxidation is caused by photosensitizers and ultraviolet light.

Research by Edelson et al. (1911? N, Eng, J, of Mad6. Vol. 318, pp. 297-303) describes certain types of cutaneous lymphoma and leukemia. An effective treatment has been described in which a photosensitizer is added to the blood and The blood is exposed to ultraviolet light.

In this study, the aromatic compound, 8-methoxypsoralen orilen) was administered to the patient. After a few hours, the blood is removed and exposed to ultraviolet light. Light (type A) was irradiated. This blood was then reinfused into the patient. but However, the mechanism of this treatment method was not explained in Edelun's study, and it is simply It has only been shown to be effective against certain types of cancer.

The present invention provides several methods to oxidize fatty proteins, and photooxidation is a method for oxidizing them. It's one of them. Adding a photosensitizer such as 8-meteocypunlaren to the blood , then irradiating the blood with ultraviolet light produces oxidized fat proteins. considered to be a thing. Furthermore, oxidized fat proteins are also considered to be effective as therapeutic agents for diseases. available.

The chemical formulas for the three types of oxidation are as follows:

Autooxidation: 0. ---1>02-111>Product peroxidation: H-0- 0-H--->H-0-0'+H'---->Product +HC Photooxidation: Photosensitizer + High voltage (IIV) -----> [Photosensitizer]'+ Oyu −−−−＞　o4　＋　Hゆー　−−−−＞Woo’　−−−−)HC− ---> From product experiments, the rate of autoxidation is determined by the concentration of the substance and the amount of oxygen above It was found that it is directly proportional to the partial pressure. Furthermore, this ratio depends on the degree of oxidation. This is indicative of a chain reaction and autocatalytic nature of the process. first The main products are hydroperoxides. Figure 1 shows these relationships, where where 0 and λ are experimental constants, RH is a non-oxidizing substance, and po is the partial pressure of oxygen. , and the rate of oxidation is expressed as the time derivative d/dt.

To date, most of the research dealing with the oxidation of fatty acids has focused on lyleate (linols). ite) is concentrated on autoxidation. Lithium chloride, a fatty acid that occurs naturally in the animal body. Ruate is a good model for the mechanism of oxidation.

As shown in Figure 2, highly purified polyunsaturated fatty acids have a long lifespan in the presence of oxygen. The first reaction is particularly important as it has been shown to be stable for a long time (Bribet To and Blank, 1962). as a catalyst (purple with traces of peroxides or transition metals) It is known that outside radiation or ionizing radiation, as well as some other elements, are necessary for initiation. It is. Once started, the reaction begins with resonance-stable free radicals (which rapidly react with oxygen). This is continued by a chain mechanism involving the formation of peroxide groups.

This chain mechanism causes a change in the substance when the peroxide group is converted to hydroperoxide. It starts a new chain by slowly withdrawing a hydrogen atom from a molecule.

Hydroperoxide decomposes under the tube formed by homolyse and starts a new chain. gives rise to a group that can be Additionally, hydroperoxides are moderately strong oxidants. agents, including thiol-containing proteins, glutathione, and thiol-containing proteins, and especially Glutathione-dependent cytotoxicity apparently acts as a cellular anti-oxidant prophylactic. converted by the element. If not converted to alcohol, hydroperoxy The metal is usually subjected to homoliths catalyzed by transition metal ions, as shown in Figure 3. As shown in Figure 4, depending on the oxidation state of the metal, hydroperoxide or alcohol Forms a group. As shown in Figure 3, the seven alkoxy groups formed are usually hydrogen atoms. Either of the sensitive molecules in the vicinity is removed by subtraction and generation of another group. It is easy to react with other substances and initiate further reactions.

In fact, the bad odor of fats due to oxidation is due to this reaction, and this odor is due to unsaturated fats. This is the odor of unsaturated aldehydes produced by this method. For example, oleic acid The hydroperoxides formed by autoxidation are thus decomposed, as shown in Figure 5. produce aldehydes as shown.

Further reactions that the alkoxy 7 group undergoes are shown in Figures 6A-6D. Figure 6A is new An overview of the hydrogen atoms that initiate new chains is shown. Figure 6B shows the imbalance with another group. ing. FIG. 6C shows a bond, and FIG. 6D shows that there is an additional double bond, Because of this strong arrangement of double bonds of the type normally found in fat peroxidation products. becomes a polymer product.

A membrane-like structure in which fat is arranged in an orderly manner to facilitate growth responses. The reason why such a reaction occurs most rapidly is due to the nature of autoxidation. It is thought that However, the complexity of such membranes is due to their peroxide deafness. A number of factors that make testing difficult and ensure that the underlying reaction can be seen. It became necessary to use such a model organization.

One such tissue is composed of a monolayer of unsaturated fatty acids absorbed in 7-lica gel. has been completed. Thus, autoxidation of sulfuric acid can be induced by small amounts of peroxide. linoleic acid disappears by an apparent first-order motion. Figure 1 The equation shown in shows that autoxidation of polyunsaturated fatty acids is a more complex movement. are doing. Monolayers thus exhibit different peroxidation mechanisms, and this This is confirmed because the main product is fatty acid Epogyu/. Some of these reactions Of interest because epoxides are the most powerful chemical mutagens and carcinogens. This is the end.

In another model tissue, single-lamellar liposomes are incubated with unsaturated phosphates in a buffered aqueous solution. Formed by sonication of phatidylcholine , the products of kinetic oxidation are the expected 9- and 13-hydroxyoctadecane esters. hydroperogoctadecadienoie acid ), however, with the inclusion of saturated phosphatidylcholine in the ribosome. Together, the product mixture further contains epoxide, hydroperoxide and ditrihydro. It has an even more complicated structure, including the xyacyl part and the trihydro beef 7-anru part. (see also Friedwig and Porter, 1981). Acole Plasma Raidlavi1 +) is a membranous structure containing a 5050 mixture of 18:2 and 16:O The fatty acid composition of phosphatides produces very similar fatty acid products; This means that the results from the model tissue can serve as a substitute for those that actually occur in biological membranes. It shows. Lipids, Mead, Proceedings Press, 2 New York, 83 pages, (1986).

Although the prior art is replete with techniques for oxidized fatty acids including fatty proteins, the present invention In addition to a new oxidized fat protein composition, we also developed a new method for oxidizing fat proteins. It also includes.

Summary of the invention According to the present invention, horseradish peroxidase Peroxides such as hydrogen peroxide that combine with the enzymes generated generate free radicals, which are Fat proteins, preferably low-density fat proteins, can then be peroxidized, so be.

As described above, according to the present invention, the novel oxidized fat protein composition itself as well as its Methods and devices for preparing these compositions can also be provided.

Thus, an object of the present invention is to provide a novel oxidized fat protein composition. In a first preferred embodiment, the composition comprises low density fatty protein, hydrogen peroxide. , horseradish peroxidase, and a salt solution. oxidizing agent and flavin, riboflavin, ox/dase, lipoquinodase, organic peroxide or Alternatively, it is also an object of the present invention to provide another novel composition containing sec-tert-butyl peroxide. It is a purpose. Modified fatty proteins are added to the composition to enhance the effect of organic peroxides. You can also Additionally, chemotherapy such as adriamycin and mitomine-D or photosensitizers such as 8-methoxybus psorare 7 and hematoporphyrin. A composition can also be provided. In addition, elemental oxygen and fluorocarbons A novel fatty protein containing (perfluorocarbon fluosal) We can also provide quality.

Another object of the present invention is to provide a method for preparing oxidized fat proteins.

A preferred method is to add hydrogen peroxide and horseradish peroxide to a solution of low-density fatty proteins. Enzyme is added and the sample is exposed to a carbon-13 nuclear magnetic resonance spectrometer to determine the degree of peroxidation. It is to decide. Fatty proteins can also be oxidized using other oxidizing agents.

Another method involves oxidizing fatty proteins by irradiating the specimen, which A photosensitizer is added to the specimens, and the fatty protein solution is exposed to ultraviolet light. A chemotherapeutic agent is added.

It is a further object of the invention to provide an apparatus for preparing oxidized fat proteins. Ru. One such device may be a peroxidation device containing immobilized enzymes. , in which a pump injects hydrogen peroxide. For other devices, it may be fixed. a container holding a standardized enzyme to which hydrogen peroxide is added . Additional devices may include an atrioventricular anastomosis or arterial bypass attached to the patient. A peroxidation device is attached to the anastomosis.

This patent application is based on another patent application by Elie Couty and Fossell filed on the same day as the present invention. Oxidized fat proteins are responsible for cancer, malaria, and AIDS (immunodeficiency syndrome W). It has been taught that it is useful in treating diseases such as bacterial infections. These disease-causing cells have an increased number of fat protein receptors or have absorbed fat proteins. It is characterized by an enhanced ability to Disease-causing cells are less acidic than healthy cells. found to be more sensitive to the cytotoxic effects of modified fatty proteins. It is. In this way, treatment using oxidized fat proteins has the effect of killing disease-causing cells. It is thought that there is.

Brief description of the drawing FIG. 1 is an equation representing the motion of oxidized linoleic acid.

Figure 2 shows the initiation reaction in the autoxidation of linoleic acid.

Figure 3 shows that metal ions transform hydroperoxide into alkoxy groups and hydroxyl ions. The decomposition mechanism that transforms is shown prospectively.

Figure 4 shows the formation of hydroperoxide or alkoxy groups according to the oxidation state of the metal. hydroperoxides that undergo homolysis catalyzed by transition metal ions for shows.

Figure 5 shows how hydroperoxides formed by autoxidation of oleic acid This shows whether it is decomposed into aldehyde.

Figure 6A shows the abstraction of a hydrogen atom to start a new chain.

Figure 6B shows the disparity with another group.

Figure 60 shows the bond.

Figure 6D shows that a double bond has been added, which allows for the production of fat peroxidation products. can result in a strong sequence of polymeric products of the type commonly found in

Figure 7 shows the reflex region of the C-13 spectrum of a normal human plasma specimen. vinegar.

WJ8A isolates 125.8 MHz protons from normal human plasma. The olefin region of the spectrum is shown.

Figure 8B shows peroxidase (2 g/l) and then 3 aliquots of 3% peroxide. 12 of the same plasma as in Figure 8A, in which Pl/ml (100 Pl/ml) was added hourly. 5.8 shows the isolated spectrum of the 8 MHz proton.

FIG. 9 shows a device according to the invention.

Figure 1: OA when transported by the human body in response to the malignant layer; A schematic diagram of the mechanism for producing low-density fat proteins is shown.

FIG. 108 shows a schematic diagram of a general method of treating cancer according to the present invention.

FIG. 11 shows another example of the device according to the invention for oxidizing fatty proteins in the blood supply. An example is shown.

Figure 12 shows further details of the device according to the invention for oxidizing fatty proteins in the blood supply. Another example is shown.

Description of the preferred embodiment The present invention will first be explained from a broad perspective and will be explained in detail below. broad concept Therefore, the present invention provides a novel method for lateralizing oxidized fat protein simultaneously with a new fat protein composition. and its equipment.

Fatty proteins are chylomicrons (ehylo 1erons), chylomicron residues. distillate, very low density fat protein, medium density fat protein, low density fat protein, and high density fat protein. Contains dense fatty proteins and exists in various forms in the blood. The fat of the seeds is fat It is accompanied by a specific protein. That is, the special characteristics of these two classes of biomolecules It is a protein structure with mixed material properties. So there are two main types These are transport proteins and membrane ruts. These tissues contain fat and protein. The quality is not covalently bonded, but is largely due to the polarity between the fat and protein compositions. They are held together by the mutual hydrophobic action between the sexual parts.

Plasma proteins are complex, in which fat and protein are kept in relatively constant proportions. ing. They are relatively small and have a certain spherical shape and weight, and they can be carried through blood. transports water-insoluble fats between various organs. human plasma protein occurs in four main shades, which differ in terms of sphere size and density, as shown in the table below. ing.

Main classification of proteins in human plasma Kiromi Ultra-low density Low-density lipo High-density Ribochron Fat protein Fat protein Fat protein (VLDL) (LDL) (HDL) density, gl-' < 0. 94 0.94-1.008 1.006-1.063 1.063-1.21 Buoyancy rate, Sr < 400 20-400 0-20 (sedimentation) sphere size, n m 5-1000 30-50 20-22 7.5-1.0 protein, dry weight Amount % 1-2 10 25 45-55 Triangle Glisse, 11G-95 55-65103 dry weight% Phosphatide 3-6 15-20 22 30 dry heavy spirit% Cholesterol, free, 1-3 10 8 3% dry weight Fresherol, 2-4 5 37 15 esterified, dry weight% Passages are provided within the body to allow for interconversion between these four major categories.

As shown in the table above, plasma fat proteins are proteins with varying proportions. Contains different types of fat. Very low-density fatty proteins have a pronounced amino acid configuration. It has four different types of polypeptide chains with columns. -Blade high-density fat Fatty proteins come in two different types: molecular weight 17,500 and min+1cor, ooo. It has a polypeptide chain. The polypeptide chains of blood plasma fatty proteins are the surface of the molecule. They are thought to be arranged in a plane, and therefore their hydrophilic properties are being compared. But long In extremely low-density fatty proteins and chylomicrons, there is insufficient protein to cover the surface. The type of phosphatide composition is internal non-polar triadylglycerol. It is thought that the molecule also acts as a hydrophilic group on the surface. biochemistry , Lehninger, Worth Publishers, Inc. 1, New York, 1975. See page 301.

According to a most preferred embodiment of the invention, the oxidized fat protein composition comprises low density fat protein. Contains white matter solution, hydrogen peroxide, horseradish peroxidase, and salt water . In all of these examples, the term ``fatty protein solution'' refers to saline or buffered solutions. Must be understood as either fat or protein. In addition, fat and protein The fluid consists of low-density adipose protein, high-density adipose protein, or very low-density adipose protein However, a fatty protein solution containing low density fatty protein is a preferred embodiment of the invention.

In another embodiment, the composition of the low density fat protein solution includes flavin, riboflavin, Oxidase, peroxidase, horseradish peroxidase, lipoxidase, ver Any acid selected from 2#-nido, organic peroxide, or tertiary butyl peroxide Contains a curing agent.

Preferred methods for preparing oxidized fat proteins 1. In a specific example, the solubilization of low-density fatty proteins It involves adding hydrogen peroxide and horseradish peroxidase to the solution.

In this method, oxidized fat protein is then injected into the patient. degree of peroxidation To determine and monitor the degree of .

In another embodiment of the method according to the invention, fatty proteins naturally occurring in the patient's body oxidizes quality. Hydrogen peroxide itself or e.g. hydrogen peroxide or pokindase Hydrogen peroxide combined with enzymes such as It is administered into the patient's body by injection.

Administering non-oxidized modified adipose protein is important when formulating oxidized adipose protein. Makes the monster's effect stronger. Modified adipose protein is administered secretly before oxidation of adipose protein. or added to the solution. Modified fatty proteins are, for example, t’)’J/Ii FulltlJgu’J Se! J I’ (tri 1lnolea triglyceride, silyl alphospha Cholesterol, such as tinorcholine and cholestecool ester of linoleic acid It is formulated by concentrating the natural fatty protein content with esters. these substances that oxidize more readily, resulting in more cytotoxic peroxidation products. Conceivable. Modified fatty proteins are included in each embodiment of the invention.

The body's fat peroxidation process further inhales increased levels of elemental oxygen during breathing. It can increase the oxygen level in the blood and promote the effect. bellefluo Rocarbon Fluosal is also a drug that increases oxygen levels in the blood and can be injected intravenously. more injected into the body. Oxygen levels in the blood depend on whether oxidized fat proteins are administered or Or it increases before oxidation in the body. These two increase oxygen levels in the blood Both methods are included in each of the embodiments of the present invention.

In all of the examples described here, the blood has a degree of oxidized fat protein. monitored with a carbon-13tIF magnetic resonance (NMR) spectrometer to determine There is.

This measure was filed on July 24, 1990 by Erickty, Fossell. U.S. Patent Application No. 557,529, which includes Cancer by measuring fat peroxidation using nuclear magnetic resonance (NMR) spectroscopy A method of detection is described. In cancer survivors, fat peroxidation occurs and polyvalent It was found that the ratio of saturated fatty acids to monounsaturated fatty acids was different in healthy people. ing.

Nuclear magnetic resonance (NMR) spectroscopy values for methyl and methylene groups in fat are: Determined by comparison with corresponding values for healthy people.

In the examples described herein, fatty proteins are used as a standard for whole blood, plasma, and serum. Peroxidized by books.

In the apparatus according to the present invention shown in FIG. is directly peroxidized. The peroxidation device or peroxidation chamber SO may contain, for example, peroxidase or has an immobilized enzyme 52 such as lipoxidase and an inlet 54 from the pump SS. This pump 55 provides a very gradual and precise flow of hydrogen peroxide into the device. It is injected. The patient's artery through which the blood flows is connected to the peroxidizer via the inlet 53. It is injected into. Blood containing peroxidized fat proteins is discharged from the patient through the spout 51. returned to the vein.

FIG. 11 shows another embodiment of the invention, in this case blood drawn from patient 57. 56 is stored in a container 58. Beroxide 59 is added to another container 60 and this content is It is stored in the container 60. Beroxide 59 is then added to container 58, thereby Fatty proteins in the blood are oxidized. Blood containing oxidized fatty proteins61 is the It is then injected into the patient 50 again.

FIG. 12 shows yet another embodiment of the invention. Heparinized blood 66 is The container 68 is coated with horseradish peroxidase, for example. It holds an immobilized enzyme, such as a membraned sphere 70. Hydrogen peroxide 2 is at the bottom of the container. As a result, it becomes oxidized fat protein 74 in the blood and reaches the top of the container. It is poured out from the department. Blood 74 containing oxidized fat proteins is stored in a container 75 and its It is administered to patients when treatment for later illnesses is indicated.

Another method of oxidizing fat protein according to the present invention is to add a photosensitizer to the fat protein solution, Thereafter, the method includes the step of exposing the solution to ultraviolet light. 8-Metoo beef/Busora By irradiating the molecule, an intermediate group is generated, resulting in the generation of a hydroxyl group. next These groups cause peroxidation of fatty proteins, which is effective in treating phosphorus cells. It gives. Blood is thus removed from the patient and the blood contains 8-methoxychloride. Sibuspsoralen is added and the specimen is then exposed to ultraviolet light, thereby This results in peroxidation of the fatty proteins inside.

Hematoporphyry used in the same manner as 8-methoxybusoralen described above It is also known that cancer cells can be killed in irradiated blood. . According to the invention, 8-methoxybusoralen, like hematoporphyrin, It is thought to be effective in peroxidizing fat proteins.

According to the present invention there is a further method effective for the peroxidation of fatty proteins.

This method involves adding a chemotherapeutic agent to a solution containing fatty proteins.

Atriamine (AD) and other molecules in its class are effective against several types of cancer. It has long been used as a chemotherapeutic agent. They also generate free radicals It has been thought that The mechanism of such a reaction is unknown, but it is possible Two mechanisms are shown below.

1) AdH2+Ou −−−−−−> AdH+ O, −+ H”AdH+o 2−−−−−>　Ad　　　　o；　+　H“Enzyme 2) AdH Yu　　　　－－－－－＞　A　dH+　O:,″+　H+3+ Other metal ions can also be used in place of Fe. Furthermore, as a chemotherapeutic agent Might Mine D is in the same class as Atria Mynon (mito yein- D) can be used interchangeably. In this way, according to the present invention, the atrium Amainon and mitomacin-D generate free radicals, which then oxidize fat proteins. give rise to

In another patent application filed on the same day as the present application, Erick, Tee, Fossell , oxidized fatty proteins are involved in diseases such as cancer, malaria, and bacterial infections such as AIDS. It teaches that it is effective in treating certain diseases. Dead cells of these diseases have an increased number of fat protein receptors or an increased ability to absorb fat proteins. It has the characteristic of being strengthened. When low-density fat protein is oxidized, this protein is a cytotoxin that selectively kills phosphorus-producing cells that have an enhanced ability to take up fatty proteins. It has the effect of As part of the response to diseases such as cancer, The body oxidizes fat proteins and circulates them in the blood. phosphorus source cell are more susceptible to the cytotoxic effects of oxidized fat proteins than healthy cells. It has been found that In this way, treatment using oxidized fat protein It is thought to have the effect of killing phosphorus source cells.

Figure 1OA shows how peroxidized low-density adipose protein cytotoxicity causes cancer in the human body. It shows that it is useful in combating. Generally, cancer cells 30 contain tumor necrosis elements (tumor necrosis elements). Macrophages that secrete necrosis factor (TNF)34 32.

Tumor necrosis factor (TNF) 34 is a polymorphonuclear neutrophil leukocyte (polysorphon uclair nauttrophlls) (PklN) 31 and respiratory failure. It cracks and releases superoxide Of°. Superoxide has a composition of hydroxyl free radical “OH” (This hydroxyl free radical in turn oxidizes low-density fat protein 33) Hydroxyl ion Peroxidized low-density fat protein (P-LDL) 35 while being converted to ``0■ That's what I do. Peroxidized low-density fatty proteins then have a cytotoxic effect on malignant cells36. This causes the cells to die. Low-density fatty protein (P-LD) L) The malignant cells 36 killed by 35 are the tumor cells detected from the beginning. There are times when it is the same as 30, and times when it is different.

FIG. 108 illustrates a method according to the present invention. Low-density fatty protein [33 is peroxidized Irradiation of agent 40 directly converts it into peroxidized low-density fatty protein (P-LDL). will be replaced. In one embodiment, peroxidant 40 is peroxidant with It is a chemical enzyme. Malignant cells 36 are converted to peroxidized low-density fat protein (P-LDL). It can be killed by irradiation.

The invention is further described by the following examples, which are not intended to limit the invention. isn't it.

Example l Oxidized low-density fat protein was prepared from Sigma Chemical Co., catalog number L2139. prepared from low-density fatty proteins available from Prepare it from plasma in the usual way. Lindren FT, Nrubazuei, Jutangerar, Ray/1. fat by y toel, bluff dolly diddy (Lipids) 1977; 12: 278-282, and Lindren. By F.T., Adamson G.L., Jen Lin L./-, Woodbeedy. Lipids 1975; 10: 750-756 #Steru.

Oxidation of low-density adipose proteins is accomplished using soluble horseradish peroxidase (Enzyme Committee Classification table, No. 1.11.1.7) or immobilized carbon dioxide as a catalyst. This was done using one of the wasabi peroxidases.

The immobilized enzyme can be separated from the oxidized low-density fat protein solution before use. It has characteristics. each -1 (most preferably 5B per -1) of low-density fat protein solution protein) is the same amount of Dalbe 2. Dulbeeco's phosphatide Peroxidase in either form is diluted with salt and added to each part of the solution. It was added at a level of approximately aoo-too units. Next, hydrogen peroxide, i, o, most preferably 0.1 ml of 3% hydrogen peroxide is added to each added to white matter solution.

This solution was kept at room temperature and 0.1 ml of -1 low density fat protein solution was Berokind solution was added hourly for 2 hours. The degree of peroxidation depends on the carbs of the solution. Intensity ratio of resonances of 128pp■ and 130pp■ in N-13 nuclear magnetic resonance spectrum was measured.

A lower intensity ratio may result from a decrease in the amount of polyunsaturated fatty acid side chains in fat protein fats. and The intensity factor of 12B/130 pp- is usually greater than 0.9 before peroxidation. After peroxidation, it is between 0.7 and 0.85.

Example 2 Horseradish is a peroxidized low-density fatty protein. peroxidase (perox 1dase) type n (most preferably 2 sg/ human low-density fat protein (most preferably as described in Example 1) 5 g of protein/l). This peroxidation The low-density fatty protein obtained is mixed with about 70 to 200 P liters of hydrogen peroxide (most preferably or 3% hydrogen peroxide in equal aliquots of 1 or 2> and then first The mixture is prepared by adding a second dose several hours after the first addition. The degree of peroxidation is in Example 1. As described, the Carbo-13 nuclear magnetic resonance spectrum of the solution shows 128 pp■ It was measured by the resonance intensity factor of 130 ppm.

Although this technique can be employed with any adipose protein, low-density adipose proteins are peroxidized. It is particularly desirable that

Figure 7 shows the olefin region of the spectrum from a normal plasma specimen. Ru. Intensity of the peak at +28-129 relative to the peak at 1.30-131 rates are similar.

Figures 8A and 8B show two 125.8 MHz protons from normal human blood plasma. The olefinic region of the spectrum of heavily bonded carbon-13 and then peroxidase ( Most preferably, 2g/sq lid and 3 aliquots of 3% hydrogen peroxide (1 (lo%l) /Il+ was added hourly to reflect the same plasma. Ru. The intensity rate of 12B/130 basically decreases during the second blood clot treatment. However, this indicates that overoxidation is occurring.

Example 3 Oxidized fat protein is converted to 8-methquinbusoralen using low-density fat protein (LDL). It was prepared by reacting. Humans with fresh low-density fat protein (LDL) was isolated from plasma and then dialyzed overnight to remove EDT^.

Fat concentration was monitored using carbon-13 nuclear magnetic resonance spectroscopy and low density fat protein The quality (LDL) preparation was then stored at 4'C and used within two weeks. Mitsumasa The perceived response was 540 Pl (4-5-g protein/so 1) and 6041 8 - carried out using stock solutions of methoxypsoralen. 8-methoxypsora The final concentration of Ren was Zoo ng #+l. This reaction is 27-30' This was done by stirring for equilibration for 30 minutes before irradiating with C. six n's Irradiate the specimen with the Lubania FTI5TIZ PLII’A lamp for up to 6 hours. Volume reduction during injection was completed every 30 minutes. Carbon-13 nuclear magnetism of all specimens Resonance spectra were obtained with 8-methoxy7psoralen both before and after irradiation.

These spectra are due to the fat photosensitized response of low-density fat protein (LDL). used to monitor and characterize the chemical fecalization induced by these reactions. It's on.

The control plasma was irradiated with ultraviolet A for the next 30 minutes after the t2g/13Opp session. There was no change in the intensity rate of carbon-13, so the intensity rate was 0.98 before irradiation. The latter was 0.97). However, 8-methoxy7psora containing plasma The rate decreased from the value 0.98 before irradiation to the value 077 in the next irradiation treatment. and showed. Thus, the result is that the free radicals induce the oxidation phenomenon that should occur, and that the edelso This shows that the therapeutic effects observed by Ng et al.

Example 4 Oxidized fat 1 white matter was added to 6 aliquots of normal blood and adriamycin, 9 5% of 0. : Prepared by intermittent bubbling with 5% CO□. same The same amount of adriamycin was also added to another 6 aliquots of blood; Similarly, foaming was carried out using 95% N2:5% 001. 12B/130pp- The intensity ratio of the resonance changed as shown in the table below. Lower intensity rate compared to control specimens indicates that peroxidation has occurred. Ariamycin is a fatty acid when oxygen is present. Although it helps with fat peroxidation, it does not occur when oxygen is not present.

Intensity 128/130 ppm control rol pls gsa) o, as +/-o, os (n m s > control blood Sho + Adriamycin + 02 0.72 + / Q, 09 (n-16) system Blood plasma +7 F'/7 T-7in + Nx Q, 94 +/-0,0 5 (n 118) Example 5 Sec-tert-butyl peroxide was administered to humans by intravenous injection (1,v,). oxidation The degree of adipose protein was monitored by nuclear magnetic resonance and then tertiary butyl peroxide supply has been adjusted. Oxygenation of the patient's blood is achieved by administering ditributyl peroxide. Before inhalation of elemental oxygen or intravenous injection of perfluorocarbon Fluosal ( 1,v,) was also increased.

Triglycerides, phosphatides, or Intravenous injection (i, v) of fatty proteins concentrated with cholesterol esters ,) increased the supply of fatty proteins to the patient.

In this method, tertiary-butyl peroxide contains riboflavin, peroxidase, and lipophilic acid. of kindase, or other flavins, peroxides, organic peroxides, or oxidases. You can replace it with an appropriate amount.

Example 6 A ventricular-atrial anastomosis (shunt), or arterial bypass, was installed in the human body. external affairs A peroxidizer device was attached to the ventricular anastomosis (AV) or arterial bypass. external affairs The peroxidizer has an injection connection from the pot, and this pump pumps the hydrogen peroxide into peroxides. It is flowed into a device containing enzyme or lipokindase. Peroxidase or Ri Poxidase peroxidizes blood plasma fatty proteins in the presence of hydrogen peroxide. Ru.

A blood supply port is attached and the blood supply port is connected to the donor, blood bank, or Any other supply port may be used.

Fatty proteins are then oxidized by adding an oxidizing agent to the blood; Fatty proteins are produced.

Adding elemental oxygen or perfluorocarbon fluorosal to the blood makes it available in the blood. The available oxygen is further increased.

The fatty protein content of the blood may also be affected by triglycerides, phosphatides or cholesterol. can be increased by adding fat protein concentrated by esters. Ru.

Although the present invention has been described in conjunction with preferred embodiments thereof, various substitutions and modifications will occur to those skilled in the art. It may be possible to change it. However, all such substitutions and modifications shall be covered by the appended claims. without departing from the scope of the invention as described herein.

FIG, 4 ROOH+ M(n+1)”-+ROOa+M”+H” International Survey Report Notice

Claims (45)

[Claims] 1. A method for preparing oxidized fatty proteins having the following steps: (a) providing a specimen of a human body fluid; (b) having the specimen immobilized with an enzyme; (c) injecting peroxide into the specimen in said container; The process of producing oxidized fat protein by 2. 2. The method of preparing oxidized fat proteins of claim 1, wherein said specimen comprises whole blood. 3. 2. The method of preparing oxidized fat protein of claim 1, wherein said specimen comprises plasma. 4. 2. The method of preparing oxidized fat proteins of claim 1, wherein said specimen comprises serum. 5. Preparing the oxidized fat protein of claim 1, wherein said specimen comprises high density fat protein. Method. 6. The oxidized fat protein of claim 1, wherein said specimen comprises purified low density fat protein. How to prepare. 7. 2. Preparing the oxidized fat protein of claim 1, wherein said specimen has very low density fat protein. How to do it. 8. The oxidized fat protein according to claim 1, wherein the enzyme is horseradish peroxidase. How to mix. 9. 2. The method of preparing oxidized fat protein according to claim 1, wherein said peroxide is hydrogen peroxide. 10. The method for preparing oxidized fat proteins according to claim 1, wherein said specimen comprises heparinized blood. How to match. 11. The immobilized enzyme is a coated sphere of horseradish peroxide. A method for preparing the oxidized fat protein of claim 1. 12. Carbon-13 nuclear magnetic resonance spectrum of oxidized low-density fat protein aqueous solution Claims that the limit of fat peroxidation can be determined when the rate is measured at 128/130 ppm Method for preparing oxidized fat protein in Item 1. 13. Furthermore, certain triglycerides, phosphatides, or cholesterol esters Claim 1 comprising the step of administering the non-oxidized improved fatty protein concentrated by the method of claim 1. How to prepare oxidized fat protein. 14. Furthermore, by adding oxygen element or perfluorocarbon Fluosal, The oxidized fatty protein according to claim 1, further comprising the step of increasing available oxygen in the blood. How to prepare white matter. 15. Prepare a solution of low-density fatty proteins, including flavins, riboflavins, and oxidants. peroxidase, horseradish peroxidase, lipoxidase, peroxide, Adding an oxidizing agent selected from the group consisting of organic peroxide or sec-tertiary butyl peroxide. A method for preparing an oxidized fat protein comprising the step of oxidizing the fat protein by oxidizing the fat protein. 16. 16. The oxidation method of claim 15, wherein said fatty protein solution comprises low density fatty protein in saline water. How to prepare fat protein. 17. 16. The oxidation method of claim 15, wherein said fat protein solution comprises low density fat protein in a buffer. How to prepare fat protein. 18. A method of preparing oxidized fat protein comprising the following steps: (a) by intravenous injection; administering an oxidizing agent to a human body in an amount sufficient to produce an oxidizing effect; (b) A method for monitoring the degree of oxidation of fat proteins by carbon-13 nuclear magnetic resonance. Cheng; Also, (c) A step of adjusting the amount of oxidizing agent supplied according to step (b). 19. Oxidizing agents include riboflavin, peroxidase, lipoxidase, flavin, peroxidation 19. The oxidase is selected from the group consisting of oxidase, organic peroxide, and oxidase. How to prepare oxidized fat protein. 20. Furthermore, by inhaling elemental oxygen or injecting the perfluorocarbon Fluosal. 19. The oxidized fat protein of claim 18, comprising the step of increasing available oxygen in the blood. How to prepare. 21. Additionally, triglycerides, phosphatides, or cholesterol to ester 19. The oxidized fat protein of claim 18, further comprising the step of injecting the concentrated improved fat protein into the oxidized fat protein. How to formulate quality. 22. A method for preparing oxidized fat protein having the following steps: (a) of human body fluids; (b) adding a photosensitizer; and (c) mixing them. A process in which a compound is irradiated with ultraviolet light, thereby producing oxidized fat proteins. 23. 23. The method of preparing oxidized fat protein of claim 22, wherein said specimen comprises whole blood. . 24. 23. The method of preparing oxidized fat protein of claim 22, wherein said specimen comprises plasma. . 25. 23. The method of preparing oxidized fat proteins of claim 22, wherein said specimen comprises serum. 26. 23. Preparing the oxidized fat protein of claim 22, wherein said specimen comprises high density fat protein. how to. 27. 23. The oxidized fatty protein of claim 22, wherein said specimen comprises purified low density fatty protein. How to prepare white matter. 28. 23. The preparation of the acid fatty protein of claim 22, wherein said specimen has very low density fatty protein. how to. 29. The oxidized fatty protein according to claim 22, wherein the photosensitizer is hematoporphyrin. How to mix. 30. 23. The oxidized fatty protein of claim 22, wherein the photosensitizer is 8-methoxybusoralen. How to prepare white matter. 31. Carbon-13 nuclear magnetic resonance spectrum of oxidized low-density fat protein aqueous solution Claim in which the limit of lipid peroxidation is determined when measured at a rate of 128/130 ppm Method of preparing 22 oxidized fat proteins. 32. A method for preparing oxidized fat protein having the following steps: (a) fresh human providing a specimen of plasma; (b) separating low-density fatty proteins from the plasma; (c) adding 8-methoxybusoralen; (d) mixing them; (e) irradiating the mixture with ultraviolet light for up to 6 hours; producing oxidized fat proteins; (f) Carbon-13-nuclear magnetic resonance spectrum of oxidized low-density fat protein aqueous solution A process in which the limit of fat peroxidation is determined when the rate of 128/130 ppm is determined. 33. A method for preparing oxidized fat protein having the following steps: (a) plasma labeling; (b) administering a chemotherapeutic agent to the specimen; and (c) ) the mixture is intermittently aerated with a mixture of O2 and CO2, thereby A process that produces oxidized fat proteins. 34. 34. The oxidized fat protein of claim 33, wherein the chemotherapeutic agent is adriamycin. How to mix. 35. 34. The oxidized fatty protein of claim 33, wherein said chemotherapeutic agent is mitomycin-D. How to prepare. 36. Carbon-13 nuclear magnetic resonance spectrum of oxidized low-density fat protein aqueous solution Claim in which the limit of lipid peroxidation is determined when measured at a rate of 128/130 ppm Method for preparing 33 oxidized fat proteins. 37. A method of preparing oxidized low density fat protein having the following steps: (a) low density providing a solution of adipose protein; (b) catalyzing the oxidation of low-density adipose protein; The introduction of low-density fatty proteins into peroxides in the presence of enzyme catalysts that can (c) Carbon-13 nuclear magnetic resonance scanning of oxidized low-density fat protein aqueous solution The limit of fat peroxidation is determined when the spectrum is measured at 128/130 ppm rate. step (d) step (b) until the desired amount of low-density fat protein is obtained; A process of repeating (c). 38. A method of preparing oxidized low density fat protein having the following steps: (a) fresh Low-density fatty protein from human or animal plasma or commercially prepared plasma Preparing a white matter solution; (b) equal amounts of each of the low-density fat proteins; diluting with phosphatide buffered saline; (c) Adding horseradish peroxidase; (d) Keeping the solution at room temperature. (e) adding hydrogen peroxide every hour for 2 hours; (f) oxidizing low density Carbon-13 nuclear magnetic resonance spectrum of fat protein aqueous solution is 128/130p a step in which the limit of lipid peroxidation is determined when the pm rate is measured; (g) using the formulation; Storing at 4°C until use. 39. An oxidized low density fat protein composition having: (a) a low density fat protein solution ; (b) immobilized enzyme; (c) salt aqueous solution; (d) Peroxide. 40. An oxidized solid fat protein composition having: (a) horseradish peracid; 1 or 2 aliquots of human low-density fatty protein (b) containing enzyme type II 3% hydrogen peroxide. 41. An oxidized low density fat protein composition having: (a) fresh human or animal Plasma or a commercially prepared low-density fatty protein solution from plasma; (b) phosphatide buffer salt; (c) horseradish oxygen peroxide; and (d) hydrogen peroxide. 42. A device for oxidizing fatty proteins with: (a) peracid containing immobilized enzymes; conversion device; (b) an inlet from a pump; and (c) A pump capable of accurately loading hydrogen peroxide into the device. 43. A device for oxidizing fatty proteins having: (a) attached to a human being; (b) said ventricular-atrial anastomosis comprising an immobilized enzyme; or external peroxidation device attached to the arterial bypass; (c) an inhabitant connection from the pump to said device; (d) administering hydrogen peroxide to the device; pump. 44. 44. The fatty protein of claim 43, wherein the enzyme is lipoxidase or peroxidase. A device that oxidizes 45. A device for oxidizing fatty proteins having: (a) holding immobilized oxygen; (b) hydrogen peroxide in the bottom of the container; (c) means for draining the oxidized fat protein from the top of the container. 45. 45. The immobilized enzyme comprises horseradish peroxidase coated spheres. A device that oxidizes fat proteins.