Lysis/resealing process and device for incorporating an active ingredient in erythrocytes

A technology of red blood cells and active ingredients, which is applied in the field of cracking/resealing and devices for incorporating active ingredients into red blood cells, and can solve problems such as lack of industrialization methods

Active Publication Date: 2007-09-05
ERYTECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as mentioned by C.G. Millan et al. (supra) with reference to phytate for general oxygen transport, the use of erythrocytes for incorporation of molecules such as phytate currently suffers from a lack of validated industrial methods

Method used

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  • Lysis/resealing process and device for incorporating an active ingredient in erythrocytes
  • Lysis/resealing process and device for incorporating an active ingredient in erythrocytes
  • Lysis/resealing process and device for incorporating an active ingredient in erythrocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Example 1: Device

[0120] Refer first to Figure 1. The first dashed box indicates the first module 1 , generally parallelepiped-shaped, comprising a glass front (which is not shown), which can be opened and closed. Peristaltic pumps P1 , P2 and P3 are located at the bottom of the module as well as receivers (not labeled) for removable components which will now be described. Pumps P1 and P3 have constant, preset flow rates. Pump 2 is controlled to vary the flow rate.

[0121] The removable component comprises a flexible pouch 2 containing a suspension of red blood cells to be lysed. The bag 2 is accompanied by a flexible tube 3, ring-shaped, which cooperates with the pump 1 to circulate the pouch to keep the red blood cells in suspension. The bottom of the bag is further connected to a flexible tube 4 which is connected to the inlet of the "blood" compartment of the dialysis cartridge 5 . Tube 4 cooperates with pump 2 to circulate the suspension from the bag to the...

Embodiment 2

[0133] Example 2: Encapsulating Asparaginase

[0134] In this example, osmotic fragility is defined as the concentration of NaCl expressed in g / L that causes about 50% hemolysis.

[0135] 1) Effect of asparaginase on osmotic fragility:

[0136] a. Prepare asparaginase solution:

[0137] Inject 2.5 ml of 0.9% NaCl through the septum into the vial containing 10000 IU powdered asparaginase via syringe. The mixture was stirred until dissolved, thus obtaining a stock solution with a concentration of 4000 IU / ml. The contents were withdrawn with a syringe and placed in 5 ml of lysate. Prepare three solutions and store at +4°C: 0 IU / ml solution (constitutes 0.9% NaCl control), 3200 IU / ml solution (add 625 μl 0.9% NaCl to stock solution) and 1600 IU / ml solution (take 1 ml of 3200 IU / ml solution, Add 1 ml of 0.9% NaCl solution).

[0138] b. Wash red blood cells:

[0139] - starting from whole blood taken on dextrose citrate phosphate and centrifuged at 1000g for 20 minutes at +4°C...

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Abstract

Lysis / resealing process for incorporating an active ingredient, the process comprising the following steps: (1) - placing a globular concentrate in suspension in an isotonic solution having a haematocrit level which is equal to or greater than 65 %, with refrigeration at from + 1 to + 8 DEG C, (2) - measuring the osmotic fragility based on a sample of erythrocytes from that same globular concentrate, preferably on a sample of the suspension obtained in step (1), (3) - lysis and internalisation procedure of the active ingredient, inside the same chamber, at a temperature which is constantly maintained at from + 1 to + 8 DEG C, comprising allowing the erythrocyte suspension having a haematocrit level which is equal to or greater than 65 % and a hypotonic lysis solution which is refrigerated at from + 1 to + 8 DEG C, to circulate in a dialysis cartridge; the lysis parameters being adjusted in accordance with the osmotic fragility previously measured; and (4) - resealing procedure carried out in a second chamber at a temperature of from + 30 to + 40 DEG C by means of a hypertonic solution. Figure 1.

Description

technical field [0001] The present invention relates to a method that allows the implementation of the so-called lysis / resealing technique, which allows the incorporation of active ingredients into erythrocytes. The invention also relates to a device by which the method can be carried out. [0002] The present invention further relates to an erythrocyte composition containing asparaginase, etc., or phytate, and said composition can be obtained by carrying out the method according to the present invention. Background technique [0003] Lysis / resealing techniques are described in patents EP-A-101 341 and EP-A-679 101. The latter describes a relatively complex device that first comprises a cooling structure for the lysing part, in which are placed single-use components, including cartridges, tubes, chicanetype or curved ( serpentine) pouches and removable metal elements, such as curved cooling devices; followed by structures for reseals that provide reheating devices that hou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P35/02A61P35/00A61K38/50A61M1/36A61K31/6615A61K9/50A61K35/18
CPCC12M47/06A61K9/5068A61K31/6615A61K35/18A61P35/00A61P35/02A61P43/00A61P7/00A61K9/50A61K38/50
Inventor 扬·戈德弗兰
Owner ERYTECH PHARMA
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