TW449480B - Pharmaceutical composition of fibrinogen particles suitable for intravenous injection - Google Patents

Abstract

The present invention discloses a pharmaceutical composition of fibrinogen particles suitable for intravenous injection. The composition comprises a combination of human albumin and fibrinogen. The human albumin is made into a human albumin suspension according to a specific method, which is then cross-linked with fibrinogen. The composition particle will not coagulate in a suspension and has a diameter smaller than 1 micron-meter. The protein particle composition, through intravenous injection, will not cause a blockage in the blood vessel. However, it will promote the blood coagulation of a wound, and can effectively reduce the loss of blood of a patient.

Description

V. Description of the invention ⑴ '--- _ This invention relates to a medical person who can reduce the surgical and medical treatment and accelerate the medical treatment. Protein 2 hemorrhage refers to a kind of fibrinogen particles suitable for intravenous injection. It is well known that sufficient human blood is the foundation of health. Many medical treatments, such as cancer surgery, can cause blood loss. Then the blood transfusion process is very complicated, which may cause AIDS and hepatitis, especially for patients with platelet deficiency. Excessive blood loss is even more severe. Therefore, there is an urgent need for a clinical description of these problems to help with coagulation. Small liver formation As these substances have the same efficacy as platelets, they can reduce blood loss in patients undergoing surgery and surgery and patients with platelet deficiency, and their properties must be met after injection into the iliac crest. First: the diameter of human microvessels is 7 microns, then the diameter of such substance particles must be less than 7 microns; second: the substance particles themselves should not agglomerate in the bloodstream, but they can agglomerate at the wound, this substance particles will not cause blood cells Coagulates in blood vessels, but participates in promoting blood cell aggregation in the wound. Such particles of matter help blood coagulate, so these particles must have surface molecules, such as fibrinogen, that allow them to participate in blood coagulation. In current technology, such as K, I _ W dder and others (in the development of pharmacology and chemotherapy) 'V 0 1. 1 6. 2 1 3-2 71 " Anti-cancer agent biophysical illustration Magnetically Active Microspheres and Other Carriers: In this article, protein microspheres are synthesized by heating or polymerizing an aqueous solution of protein with an aqueous solution of oil and oil. However, the microparticles have a very short half-life due to the protein's extreme deterioration during this process. But in actual treatment, it needs to be able to be stored in the blood vessel for at least one day.

Page 4 4 49 48 Ο

The above will not deteriorate particles.

Oppenhein et al. In US 4,101,288 " Ultrafine particles effective in injection and its production method 提出 proposed another compound in which the use of an opposite solubilizing agent to adjust the desolvating agent to make protein The solution reached a desolvation limit. This method has two conditions, but the conditions are extremely unstable at the beginning of the reaction, and a large number of polymerizations are generated in the middle of the reaction with the three problems. The diameter of the condensed sphere is greater than 7 micrometers, which has passed the hemodiameter diameter, which can cause vascular occlusion and cause danger. Because of the actual treatment, it is required that it will not agglomerate and its diameter is much smaller than 7 microns. An object of the present invention is to provide a protein globule in a suspension, which is non-agglomerated, has a diameter smaller than 1 micron, and contains a fibrin protein composition on a surface thereof, and a production method and use thereof. In the present invention, the average diameter of the spheres formed in the presence of the crosslinking agent is less than 1 micrometer and the coefficient of variation is small. In the present invention, 'fibrinogen can be used as a longitudinal molecule, which can carry protein spheres to the wound to reduce the amount of bleeding. In the present invention, since the function of this composition is coagulation, and its efficacy is the same as that of platelets, it is also referred to as " coagulation plate " or A platelet substitute " In the present invention, the composition of this protein composition is (in parts by weight, human albumin: 90-100 parts, fibrinogen: 0.02-6 parts, and the production method of the composition of the present invention is:

449480 V. Description of the invention (3) 1. First, the protein particle suspension is prepared, where: a. The protein powder is dissolved in a buffer with an appropriate osmotic pressure; b. A surfactant is added to the protein solution described in a; c. Add a cross-linking agent to the protein and surfactant mixture described in b. d. Add the alcohol solution to the protein, surfactant and cross-linking agent described in c. Immediately, a cloudy protein sphere suspension appeared, which was suitable for intravenous injection and did not block blood vessels. In the present invention, the protein solution is a human albumin solution. In the present invention, the surfactant is a detergent. In the present invention, the surfactant may be selected from one of Sodium tetradecyl sulfate 'Tween-80' TritonX-151: In the present invention, the cross-linking agent molecule includes glutaric acid Covalent small molecule of aldehyde (glutaraidehyde). In the present invention, the P Η value of the buffer solution ranges from 4 to 8. In the present invention, the reference osmotic pressure range is 14-680m0sm. 2. After the above-mentioned suspension turbidity appears, the 'surface protein is added to the sphere in the suspension, so that the unreacted portion of the' sphere will be covalently bonded to the surface protein, which can be retinoprotein. 3. The above-mentioned sphere suspension of human albumin added with the surface protein is further processed into a powder. The process includes dilution, filtration, dialysis, refiltration, centrifugation, electrophoretic separation, and column separation.

449480

V. Description of the invention (4) Freeze-drying and powder reconstitution after freezing. In this process, τ is slowed down.

Wash the solution or change the buffer solution and add biocompatible solvents such as preservatives. The type and amount of K u W ^ are well known to those skilled in the art. Therefore, 'a pharmaceutical composition of fibrinogen particles suitable for intravenous injection according to the present invention', in which the protein particle suspension composition can cause vascular occlusion (except at the wound site), the protein particles are passed through human albumin Cross-linked fibrinogen formation with an average diameter of less than 7 $ Secondly, the present invention is also a Lengkang protein particle composition suitable for intravenous injection after reconstitution in a biological phase solution, wherein the composition ^ will cause vascular obstruction (Except at the wound site), the protein particles are formed by human albumin cross-linked fibrinogen, and the average diameter is less than $ = the particles are suspended inside the covalent form of the wound. The part of the group is formed outside the granules of the human body. The injection of blood vessels and fibers into the vein is a fibrous invention compound. In addition, the particles, which are also common proteins, are covalently strong in the wound, and the inside is a suitable static compound. The human body is made albumin subsurface average fiber diameter of less than one kind of biological particle composition by the outer portion), human albumin table form particles injected into a vein vascular resistance fibrinogen and fibrin * 7 micro requirements. Phase solution, in which the composition surface, and which are straight protein plugs (except for white molecules to molecules, after reconstitution the composition is the human body the particles are less than

Page 74944B "____ V. Description of the invention (5) Another aspect of the present invention provides a new use of the protein particles after intravenous injection into the human body, that is, the particles can migrate to new wounds and help blood To reduce blood loss in patients. In addition, the particles can be used on patients who have suffered trauma, surgery, or large, internal or external bleeding, which is extremely beneficial for reducing the blood loss and saving patients' lives. The present invention is further described below through examples. Although the following examples use human albumin, other human proteins such as immunoproteins, myoglobin, collagen, and gelatin can also be used and can form spheres. Example 1 Method for forming microparticles without fibrinogen: Put human albumin powder into a 0.9% sodium gas solution, and add J Chen degree at 0.05 mg / m 1-1 5 mg / m 1 surfactant, such as sodium tetradecyl sulfate, Tween-80 or TritonX-151, to form a protein solution with a concentration of 5-20% by weight. Then, 2 m 1 of a series of test tubes containing 2 m 1 of the above-mentioned protein solution containing different concentrations of a surfactant was added with 2 m 1 diluted with a gasified sodium solution, and the concentration range was 0.03 3% glutaraldehyde. After 10 minutes', 10 ml of ethanol [released with water to 8⑽ (volume ratio)] was added, and the solution immediately became cloudy. After another 10 minutes, dilute with 5 times the volume of the sodium carbonate solution to observe whether the suspension is clear. Xiao Guo was negative. Thereafter, the suspension was observed under a microscope at a magnification of 1000, and it was found that the diameter of the particles in the suspension was less than 1 micrometer.

At the same time, it was also found that TritonX-151 and TritonX-151 with AA T · + V 叩 / Zhangma 1-5mg / ml and

449480 V. Description of the invention (6) A mixture made of albumin with a concentration of 100-200mg / ml has the best effect. In addition, it was found that a mixture made of sodium tetradecyl sulfate at a concentration of 12 mg./ml had the smallest coefficient of variation of the particle diameter. Example 2 Method for attaching fibrinogen to protein particles as a tracking particle: The same conditions as in Example 1 were used, and human albumin labeled with 1 to 25 was used. The spheroids were pelleted by centrifugation, and the number of radioactive protein particles contained in the pellet and the supernatant was measured. It was found that 95% of the albumin particles formed spheroids. 0 9% 的 气 If the solute of the solution is doubled from 0.9% of the concentration of sodium vaporized solution, it becomes 1.8% sodium chloride solution or the 0.9% sodium vaporized solution is reduced to 0.9% The sodium sulphate solution was found to have little effect on the spheroid yield. If the pH of the buffer solution is lowered below 4, it is found that the amount of alcohol required to form a spheroid is greater than the amount of alcohol required at pH 8 ', and the yield of the spheroids is slightly increased at a pH of less than 4 By the drop. Example 3 Repeat Example 2 under the most preferred conditions. The non-conductive thiocyanate is used to coat human albumin, and Example i is used. The fiber is first used at different time intervals after turbidity. In the liquid, it was found that fifteen minutes after the formation of the sphere: the suspension of the sphere can stabilize the sphere and obtain the best mixture. Another fibrinogen must be suspended from the sphere

4 49 480 V. Description of the invention (7) The floating liquid is further processed, which includes dilution, filtration, dialysis, refiltration, centrifugation, electrophoretic separation and column separation, vacuum freeze-drying and reconstruction of frozen powder Done, in which process _ need to add or change buffer or preservatives. According to McGill et al.'S article on J.Lab.Clin.Med. 1987, 109: 127- 1 3 3, platelet mold cavity can reduce bleeding time in platelet-deficient rabbits, using platelet-deficient rabbits for pharmacological tests, The bleeding time of ten rabbits treated with the 70mg / kg Busu 1 fan method was seconds, and the difference between the sodium injection solutions was 43, and the firing without the interstitial fibrous eggs caused contraction to the patient to coagulate and shot 1 m. 1 / kg of dissolution frozen seconds). Shorten the fibrinogen in the rabbit's present blood coagulation. In the ear white matter, it is used as a short bleeding time for those who have extraneous and less fibrous egg powder in the present invention. The amount of blood lost to a wounded or susceptible particle. For example, the blood loss of the particles in the suspension can not be equally beneficial. Covered spheroids (with 0.9% gasification time reduced to 115 seconds) (standard polarizer and moved to a fluorescent microscope to observe the immune-free injection (lmg / ml) of platelet deficiency, found when bleeding was found Fluorescent particles, which indicates that the daughter leads to the area of active coagulation, and surgery for platelet deficiency is effective and can feel faster.

Claims (1)

^ 94B〇_ VI. Application Patent Scope 1. A pharmaceutical composition of fibrinogen particles suitable for intravenous injection, which includes the following components: a. Human albumin solution 10-18% by weight; b. Surfactant 0 5-2. 0% by weight; c. Crosslinking agent 10-18% by weight I; d. Alcohol solution 50-80% by weight; The range of the formation pressure of the composition is 1 4-6 8 0 m 0 sm. 2. The pharmaceutical composition of fibrinogen particles suitable for intravenous injection according to item 1 of the scope of the patent application, wherein the surfactant is sodium tetradecylsulfate, TritonX-151 or Tween80. 3. The pharmaceutical composition of fibrinogen particles suitable for intravenous injection according to item 1 of the scope of the patent application, wherein the crosslinking agent is divided into covalent small molecules of glutaraldehyde. 4. The pharmaceutical composition of fibrinogen particles suitable for intravenous injection according to item 1 of the scope of the patent application, wherein the alcohol solution is an ethanol solution. Application date on page 12: Case No .: Mu (.1 decay) Category: Steam (fill in the above columns by the Bureau) Invention Patent Specification 4494S0 Chinese name for the pharmaceutical composition of fibrinogen particles suitable for intravenous injection. Name (Chinese) 1. Xu Jiaxiang. Inventor's Name (English) 1. Nationality 1. Republic of China Residence, Domicile 1. 12th Floor, No. 28, Section 2, Dunhua South Road, Taipei City (Name) (Chinese) 1. Xu Jiaxiang Name (Name) (English) 1. Nationality 1. Residence, domicile (office) of the Republic of China applicant 1. Name of representative (Chinese), 12th floor, No. 28, Section 2, Dunhua South Road, Taipei 1. Name of representative (English) 1.