INTERLEUKIN- 1 RECEPTOR ANTAGONIST (IL-lRa) PREPARATION KIT

IN CLOSED SYSTEM TECHNICAL FIELD OF THE INVENTION

The present invention relates to equipments and tools to separate blood elements and especially to separate existed factors of the clotted blood as well as factors created through existence of elements derived from the proximity of blood to foreign particles included silica compounds and chromium sulfate compounds and also relates to the form and structure of blood elements separation in polymeric bags and also relates to the structure of filtration and separation of blood elements respective to the blood factors.

PRIOR ARTS Mechanism of pain sensation in human body created through receipt of some factors and elements derived from demolition of tissue, foreign particles and liquids entry, definite hormones secretion in body or transformation of tissues. In all of these cases, nociceptors which exist in specific tissue of the body waiting to receive a specific element like an empty hand to create necessary connection to the receiver and send a neural message to the brain. By making lesion to the body, the process of releasing from damaged tissues begins and interleukin-1 enter to the mechanism of pain as a result, after interleukin 1 being placed in interleukin 1 receptor, the pain transmission cycle is complete and the pain process occurs. In the past, many attempts have been made to relieve pain, including the use of various chemical formulas as painkillers. Such drugs in different ways affect the reduction or discontinuation of the next chain. In some cases, the molecules of these substances enter to the nerve synapses and interrupt the transmission of neural messages and consequently interrupt the feeling of pain or reduce the feeling of pain in the discussed neural synapse environment. In other types of pain medication, the direct effect of the drug on the brain system is generally evident. One of the common methods in reducing the amount of chronic pain is to neutralize the elements that cause pain at the site of pain; that is, by injecting certain amounts of drug into the inflamed tissue, it will be possible to neutralize the effects of the substances or to neutralize the recipients of the pain sensation. In most cases, the body's tissue reacts negatively to the structure of chemicals or elements, and this makes it possible to feel more pain or new inflammation due to further reactions after the effects of these chemicals are reduced.

Among the previous articles and inventions, the following can be mentioned:

Interleukin- 1 (IL-1) plays a key role in the pathology of osteoarthritis or intervertebral disc degeneration/prolapsed. The biological antagonist, interleukin-1 receptor antagonist (IL-IRa), intervenes in the physiological mechanism of these diseases. IL-IRa is a naturally occurring, structural derivative of IL-1 that competitively binds to the receptor and inhibits the biological effects of IL-1. The endogenous effects of IL-IRa are anti-inflammatory in nature, opposite to that of IL-1. A minimum IL-IRa /IL- 1 b ratio of 10:1 is required to inhibit IL-1 activity.

It has been known for some years that iL-1 Ra can be synthesized by recombinant methods. However, autologous IL-IRa, like all autologous proteins that are intrinsic to the body, is advantageous because the natural post-translational modifications such as glycosylations are already present. This is not the case with recombinant proteins because they are produced in prokaryotic hosts. Stimulation of monocytes by adherent immunoglobulin G to form the interleukin- 1 receptor antagonist is described by Arend and Leung in Immunological Reviews (1994) 139, 71-78 and Moore et al. in Am. J. Respir. Cell Mol. Biol. (1992) 6, 569- 575. Andersen et al. in Autoimmunity (1995) 22, 127-133 explains that the therapeutic effect of immunoglobulin G to be observed in vivo cannot be attributed to an enhanced formation of interleukin- 1 receptor antagonist, and that the in vitro formation of the interleukin- 1 receptor antagonist (IL-IRa) by monocytes depends on serum and plasma constituents adsorbed on polypropylene. The therapeutic use of adsorbed serum and plasma constituents to stimulate the formation of therapeutically interesting proteins in therapies is not only very costly but also involves the risk of contamination with infectious particles with which the serum and plasma constituents may be contaminated.

U.S. Patent Nos. 6,713,246 and 6,759,188 to Reinecke disclose a method for producing IL-IRa which can be employed directly in the therapy without using adsorbed serum and plasma constituents. Specifically, Reinecke provides a method for producing IL-IRa in a special syringe made of glass, quartz or a plastic, the syringe being filled with blood, and incubated, to form the IL-IRa being formed. The internal structure of the Reinecke syringe consists of a special material, in particular a glass, plastic, quartz and/or corundum, the surface of which is modified with the aid of a corrosive agent (chromosulphonic acid). The syringe is filled with a patient's blood and incubated to form IL-IRa. The blood enriched with the protein is then centrifuged (to remove solid constituents such as blood platelets) and the serum containing IL-IRa is reinjected into the patient (for example, into a diseased joint of the patient). The syringe used by Reinecke has its inner surface textured by the acid and further includes glass beads to increase the internal surface area of the syringe and, thus, to provide a larger inducing surface. The glass beads, with a diameter of from 1 to 5 mm, occupy no more than 50% of the internal volume of the syringe. In Reinecke's method, the syringe is used to remove the patient's blood, to process the blood (to produce IL-IRa), and then reinject the autologous IL-IRa back into the patient. The present invention expands upon this method, to produce autologous IL-1 Ra in a more efficient manner.

An invention with US20140371048A1 publication number titled “Integrated kit for separating blood and concentrating PRP and method for extracting PRP using the same” filed in USPTO actually is an integrated kit for separating blood and concentrating platelet rich plasma through PRP, having a main body contains 2 conical storage placed on upper and lower portion and a cylindrical part which connects these two parts to each other to transfer the blood and an adjustment screw to close the blood passage from upper storage to lower storage. In this kit the centrifugation is used to separate the red blood cell, plasma and platelets which in this method, most of platelets disappears and just a few amount of them remains to reuse.

DESCRIPTION OF INVENTION

The present invention is a kind of IL-IRn production set in blood solution and separation and extraction of the mentioned cases from the separated elements of blood with the help of centrifugation method and creating different phases in the clotted blood. This kit includes a connection with cap to connect to the needle to extract the blood. After connection, the presence of a tee piece with the ability to determine the direction of blood movement, which is located in the blood transfusion tube pass way to the bag, allows the connection of a syringe to the tee piece so that after entering the needle inside the vein, the tee piece placed to collect the blood by syringe. As the syringe is pulled back, blood is transferred from the vein into the syringe. By completing the syringe capacity, the direction of tee piece can be changed and the transfer pass way from the syringe to the blood collection bag can be opened. The collected blood is then transferred into the bag by squeezing the syringe. With the help of a polymeric tube, the said connection transports the extracted blood from patient’s vein to a polymeric bag. In transmission passage, the presence of an external lock causes the flow as well as the path of likely blood leakage from the bag to be cut off. When the defined capacity of the bag is filled, the external lock is closed and blood sampling process is completed. Now by removing the needle from the bag connection connector, the cap related to the connection is also closed. In this method, the possibility of any bacteria entering the kit is eliminated and you can be sure that the set is sterile. As the blood enters the bag area, the blood clotting process begins. Inside the bag, there are some silica volumes which can be spherical or any regular geometric shape or non-geometric with non-sharp edges in a way that does not damage the structure of the blood elements. Inside the bag, there are numbers of silica balls that have a completely spherical surface. Depending on the type of required operation, simple glass balls or coated glass balls can be used. Coating of used balls is by chromium sulfate or other effective chemicals. Blood cells begin to produce IL-IRa in proximity of glass balls. When the blood clotting process is completed, the cap closed and the coagulation begins. At this time, the blood stored in 20 to 37 C during 6 to 72 hours. After the IL-IRa production process, the blood can be centrifuged in the existing bag to separate its elements. Due to the presence of IL-IRa in the plasma element, can cause the plasma to escape from skew tube that is next to the bag by removing plasma and blood cells remains and by squeezing the bag by hand or a tool to compress it and the plasma which contains cytokines escapes through this skew tube from the bag and passing from polymeric tube, the contents of the tube reach the filter in the tube passage and after necessary filtration, enters the second bag by a threaded connection. On the exit way of centrifuged elements, the presence of a breakable lock can prevent the contents of the bag from coming out during centrifugation.

When the elements exist, first the breakable lock is broken and the external lock is opened, and at the end of the transfer stage, the contents of the main bag can be blocked by closing the external lock. When the capacity of the second bag is completed, can occlude the vessel connected to the bag which contains plasma with the help of a cap when the connection between two caps is opened. Also if it is necessary, three bags can be connected to the connection simultaneously instead of one bag and also can connect three bags to the connection system consecutively when each bag is filled. Also one to three syringes equipped with a threaded connection to extract the centrifuged factors can be used in the way that by connecting each of the syringes and discharging the centrifuged phases in the main bag, the process of filling the syringe is completed and after completing the syringe, it is separated and the next syringe is connected to the system. By completely discharging the elements separated from the first bag and filling the syringes or connected bags, the connection between the first bag and defined containers capes and the set of containers is isolated to the entry and exit of pathogens. At the end of each container bag there is an outlet port connection that can be pulled by inserting the needle into the plasma discharge port and used for injection. BRIEF DESCRIPTION OF FIGURES

Figure 1 which shows the entire shape of interleukin- 1 production pack with silica balls including:

I . Collection bag for extracted blood 2. Silica Balls

3. Connection of blood inlet tube to the bag

4. Blood transfusion tube into the bag

5. Connection of the extraction pipe to the needle or cap

6. Cap 7. Connection of the outlet pipe to the blood bag

8. The breakable external lock

9. External lock

10. Liquid purification filter

II. Connection of the main blood bag to the bag collecting the extracted derivatives

12. Extracted derivatives collection bag

13. Breakable connection to connect to the syringe

Figure 2 which shows the input cap.

Figure 3 which shows the entire shape of the blood extraction bag and the manner of connection of piping to each other.