WO2007104760A2

WO2007104760A2 - Plasma lyophilisate

Info

Publication number: WO2007104760A2
Application number: PCT/EP2007/052349
Authority: WO
Inventors: Rolf Zander
Original assignee: Johannes-Gutenberg-Universität Mainz
Priority date: 2006-03-14
Filing date: 2007-03-13
Publication date: 2007-09-20
Also published as: DE102006011534A1; WO2007104760A3

Abstract

The invention relates to the use of the autologous blood plasma (autoplasma) of a patient, in an essentially anhydrous form in a container which is closed in a sterile manner, is essentially impermeable to water vapour, and has an inner volume that can contain at least one volume corresponding to the plasma end volume, for the long-term storage and subsequent production of liquid autologous blood plasma (autoplasma) for volume replacement or volume exchange therapy.

Description

Plasma Lyophilisate

The present invention relates to the use of autologous blood plasma (self-plasma) of a patient in volume replacement or volume exchange therapy.

With massive blood loss, under emergency conditions or perioperatively in the hospital, the maintenance of the volume of the blood (volume replacement or volume exchange therapy) has acknowledged the highest priority. In volume replacement or volume exchange therapy, large volumes of fluid are used, typically in the order of 2 to 3 liters, to compensate for the loss of blood volume. Only then does priority order ensure that the oxygen transport function of the blood is sufficient and then that haemostasis be ensured (Hiippala: Replacement of massive blood loss, Vox Sang 1998; 74 (Suppl. 2): 399-407). Differences of opinion exist only with regard to the question of when acute blood loss can only be treated with a volume replacement and from when blood loss with the administration of oxygen carriers (erythrocytes). For hemostasis, i. the delivery of coagulation factors to the patient, as well as for the delivery of other lost by the blood loss proteins is administered either whole blood or Plutplasma. The amount of blood plasma administered is on the order of about 200 ml, which would be considerably too low for a volume replacement.

Opinion differences exist in the discussion of which volume replacement agent is optimal to compensate for a loss of blood volume in the patient. In addition to crystalloid solutions, eg. As isotonic saline, Ringer's solution, etc., which provide only electrolytes, numerous colloidal volume replacement solutions are available, which are characterized by their content of macromolecules, wherein the colloidal volume replacement solutions both natural (human albumin of different concentration) and synthetic colloids ( Dextran, gelatin, hydroxyethyl starch).

The question of whether plasma preparations, e.g. Frozen plasma (GFP), suitable for volume replacement or not, was not clear for a long time.

The National Institutes of Health (Consensus Conference: JAMA 1985; 253: 551-553) maintain the volume expander used to date, such as the aforementioned crystalloid or colloidal solutions, for the most therapeutically safe agents. Heim et al. (Use of FFP in the operative field with special consideration of the intrinsic FFP Infusionsther 1990; 17 (Supp 2): 34-36) consider the use of preoperative autologous frozen plasma (AGFP), which obtained during a blood donation before surgery was, for a suitable supplement to conventional volume substitutes.

The German Medical Association states in its "Guidelines for Therapy with Blood Components and Plasma Derivatives" (3rd ed., Deutscher Ärzteverlag Köln 2003) that with acute blood loss, crystalloids and artificial colloids would be preferred for maintaining or securing the target blood volume, and that human albumin solutions would also be useful in the Volume Replacement can be used, but only if the possibilities of a protein-free volume replacement were previously exhausted.For this purpose, it is further noted in the guidelines that the administration of frozen plasma as a volume replacement therapy is basically not indicated.

According to the opinion of the German Medical Association, in the case of long-term plannable operations, which are associated with foreseeable large blood loss, the preoperative recovery of several units of autologous frozen plasma (AGFP) by means of plasmapheresis is a possibility perioperatively a "physiological" blood substitute transfusion, eg in case of intoxication, Frozen plasma is therefore the drug of choice, at least in exchange transfusion.

Frozen fresh plasma (GFP), also called fresh frozen plasma (FFP), comes from at least one whole blood donation, which was obtained either from a single donor or from several hundred blood donors (pool plasma). The recovered whole blood is usually centrifuged and the resulting plasma is squeezed into a transfer bag. The recovered plasma is flash-frozen at the latest after 24 h and then stored at -30 ° C.

A disadvantage of the plasma preparations produced in this way is that the deep-frozen plastic bags are easily fragile. In addition, the plasma must be brought to body temperature after thawing only with specially approved equipment, because overheating to more than + 42 ° C must be avoided. After thawing, it must be ensured that all protein precipitates are dissolved. Further disadvantages of the conventional freeze plasma are its low storage capacity, which is a maximum of 12 months, and the high storage costs, since conventional frozen plasma bags need a lot of storage space and the storage temperature must be at least -30 ° C. Due to the limited storage time, according to the Paul Ehrlich Institute only 1.5% of the 1.5 million frozen plasma preparations produced in Germany are transfused, while the remainder expire.

The object of the present invention was to provide a means for volume replacement and volume exchange therapy, which overcomes the disadvantages of known volume replacement means, is inexpensive to produce, at the same time provides the patient important blood components and thereby safe, long-term stability and easy to store and transport.

This object is achieved by the use of autologous blood plasma (self-plasma) of a patient in substantially anhydrous form in a sterile sealed, substantially wasserdampfundurchlässigen container whose inner volume may include at least one of the plasma end volume corresponding volume, for long-term storage and subsequent production of liquid autologous blood plasma (Own plasma) for volume replacement or volume exchange therapy.

The use of autologous blood plasma (self-plasma) in the substantially anhydrous form in a container according to the invention for volume replacement or volume exchange therapy has a number of advantages over previously used volume replacement agents and also against the used for other purposes, especially the supply of blood coagulation factors, foreign plasma in frozen form.

A significant advantage of the use according to the invention of autologous blood plasma in the form according to the invention for volume replacement or volume exchange therapy over known critical and colloidal volume replacement agents is that the autologous blood plasma simultaneously also contains a number of endogenous important proteins and factors such as blood coagulation factors. Thus, even in emergency treatment with massive blood loss in the priority order after the volume replacement and ensuring adequate oxygen transport function of the blood following assurance of hemostasis can be omitted by additional administration of blood coagulation factors.

Another advantage of the autologous blood plasma of the present invention in substantially anhydrous form is its long term stability to freezing plasma. The autologous blood plasma in essentially anhydrous form can persist for many years, usually more than 10 years, - -

stored and then used as a volume replacement agent. In contrast, frozen plasma can be stored for a maximum of 12 months, which is why in Germany about one third of all frozen plasma preparations have to be expired and discarded.

Another advantage of the use of autologous blood plasma in accordance with the invention in substantially anhydrous form is the possibility of storage at ambient temperature, whereas frozen plasma must be stored at at least -30 ⁰ C.

Another advantage of using autologous blood plasma (self-plasma) in the substantially anhydrous form of the present invention is the safety of the patient from whom the blood plasma originated and for whose volume replacement or volume exchange therapy it is intended. Since the blood plasma used in conventional plasma therapy usually comes from blood donations of a variety of donors, it must be before storage or

Use of extensive serology on blood group affiliation and pathogens (eg, HIV, hepatitis, etc.). This is extremely expensive and costly. In the

Use of autologous blood plasma for the invention Tw & ςk can this

Investigations are omitted and the product thus offered significantly cheaper. In addition, the use of autologous blood plasma is much safer, as well as serological

Tests are flawed and may not detect new or rare pathogens.

The aforementioned advantages of the use according to the invention, long-term stability, shelf life at ambient temperature and safety open up advantages which were not previously available.

The use according to the invention makes it possible, for example, for a person to store a supply of autologous blood plasma which can be stored for many years and, because of its stability at ambient temperature, can always be carried by the person himself. Due to the small volume, this entrainment of the plasma bag is also very comfortable. In an emergency, the plasma stock can be used immediately as volume replacement or volume exchange without serological tests are necessary or there is a risk of infection for the person.

Areas of use which are opened up by the use according to the invention are, for example, stays in areas with poor medical care with respect to blood and plasma bags and volume replacement agents. Such stays may be, for example, travel to developing areas of the world or military deployments of soldiers. If the autologous blood plasma bags are provided with the name and / or photograph of the person from whom the blood plasma originates, then in an emergency a doctor only needs to compare this information on the bag with the identity card or the face of the person and can immediately and use without loss of time as volume replacement or volume exchange means. - -

The present invention also provides the opportunity to make blood plasma now easier and less expensive for volume replacement therapy or volume exchange therapy by storing the blood plasma in substantially anhydrous form in a sterile, substantially water vapor impermeable container.

By the removal of water from the blood plasma, the volume of the blood plasma bag, which is usually stored and provided in a bag, significantly reduced, so that, for example, a plasma volume of 1 liter is reduced to the volume that a bag in DIN A5 to DIN A4 format, which contains only about 75 g of plasma solids. Accordingly, the weight of the plasma bag is reduced from just over 1 kg to the aforementioned 75 g plus the bag weight. As a result, the present invention has the advantage over conventional blood plasma bags for use in volume replacement and volume exchange therapy in that the blood plasma bags of the present invention are significantly more space efficient and less expensive to transport.

In addition, the blood plasma bags according to the present invention can be stored well over 12 months, so that by their use the conventional annual committee of 34%, which was determined by the Paul Ehrlich Institute (see above), fails, resulting in significant Cost savings result because, overall, much less blood plasma must be produced to store a certain amount of stored blood plasma over a period of more than a year.

Furthermore, the storage costs can be significantly reduced by the use of blood plasma in substantially anhydrous form according to the present invention, because the storage of substantially anhydrous blood plasma at room temperature, ie substantially uncooled, can take place, whereas the storage temperature for conventional freezer plasma bags -30 ° C must be. However, the storage of blood plasma bags according to the invention in moderately air-conditioned rooms (temperatures between 15 and 25 ° C) may be preferred.

The advantages of the present invention are particularly pronounced when the blood plasma is stored in almost completely anhydrous state, with complete absence of water here at least the degree of drying is understood, the blood plasma can reach by conventional freeze-drying or vacuum centrifugation. The use according to the invention of autologous blood plasma, ie intrinsic plasma in an essentially anhydrous form of a certain person to whom the plasma or blood has been taken to reuse this plasma for the treatment of this person, now opens up the possibility of storing one's own plasma to a large one Patient circle, without the costly cooling facilities are required.

The storage in this form can now be carried out under the simplest conditions, so that the donor / patient can keep his own blood plasma bag itself readily, for example, in a suitable place in his household. This in turn has the advantage that in principle any donor / patient is able to create his own, suitable for him blood plasma depot, which can be used if necessary, without relying on foreign plasma, which in turn a very expensive and expensive Diagnostics should be subjected to hedge the compatibility of the foreign plasma for each patient. In addition, this system also excludes the risk of infection from foreign plasma.

Moreover, due to the low weight and low requirements of the substantially anhydrous plasma, it is now also possible to carry at least some "emergency ration", for example when staying in areas with relatively poor medical care.

Due to the virtually unlimited shelf life of the blood plasma bags according to the invention for the volume replacement and volume exchange therapy, the construction of an adequate self-plasma depot, even in the extreme case, can take place cumulatively over a period of several years or even decades. According to current regulations, a maximum of 600 ml can be withdrawn via the plasma separation per single donation. The total volume of the withdrawn blood plasma per year and person can be up to 25 l (Hiller et al .: Indications for the therapy with fresh frozen plasma Dtsch med Wschr 1989; 1 14: 1371-1374). This means that over a period of several years, a young person can create a virtually unlimited volume replacement volume, which he can later refer to as a potential patient. This may be of particular interest to members of those occupational groups who are at increased risk of injury, and to those who have a blood type for whom plasma reserves are frequently not available.

The storage time for the blood plasma used in accordance with the invention in substantially anhydrous form is preferably longer than 1 year. Particularly preferred are storage times of at least 5 years or even at least 10 years. Preferably, the water is removed from the withdrawn blood plasma by lyophilization. In an alternative embodiment, the water is removed by vacuum extraction from the withdrawn blood plasma. In other alternatives of the present invention, the water is at least partially removed by lyophilization and / or vacuum extraction, whereby the two aforementioned methods can be combined either separately or in combination with other drying methods known in the art.

The blood plasma is preferably admixed after removal and before further processing with a stabilizer solution. This stabilizer solution is preferably a citrate-containing stabilizer solution. Suitable solutions and their concentrations as well as the volume of citrate solution to be added per blood plasma volume are known in the prior art. In an alternative embodiment, heparin can be added to the blood plasma instead of a citrate-containing stabilizer solution or additionally for coagulation inhibition.

In a single donation, usually 450 ml of blood are mixed with 63 ml of a citrate-phosphate-dextrose solution (CPD solution), with the blood being fed being weighed into a bag in which the CPD solution has already been introduced. Thus, with a hematocrit of 45% (typical mean), 247.5 ml of plasma (55%) is diluted with 63 ml of CPD solution, i. to a final concentration of 20.2% by volume.

If only plasma is to be obtained, the erythrocytes can be returned to the patient. In this case, 495 ml of plasma are recovered and added with 126 ml of CPD solution (20.2 vol .-%). The total volume is then 621 ml.

The use of heparin or its derivatives may be advantageous over the use of citrate solutions, since citrate solutions may have a slight alkalinizing effect on the plasma preparation. In alternative embodiments, however, it is precisely this alkalizing effect that is desired, so that again the use of citrate solution is preferred in these embodiments.

In the plasma collection today CPD solution (citrate-phosphate-dextrose solution) or ACD solution (aqua-citrate-dextrose solution) are used as stabilizer solutions for the prevention of blood clotting. According to the invention, the use of these commercially available solutions is not absolutely necessary since phosphate and dextrose (glucose) are advantageous only for the storage of the erythrocytes separated from the donation and stored separately - -

is. For the stabilization of the plasma as such, the use of a pure citrate solution is sufficient according to the invention. As a result, the plasma dilution by the stabilizer solution can also be advantageously reduced if a correspondingly highly concentrated citrate solution is used.

The stabilizer solutions (CPD solution, ACD solution, citrate solution) have the following compositions (solvent = water):

Preferably, a container is used as a container for holding the liquid, frozen or dried blood plasma, whose inner volume is variable, wherein a corresponding container preferably at least partially made of flexible material, with the proviso that the flexible material as well as the other components of the container, which are in direct contact with the interior of the container, are substantially water vapor impermeable.

"Substantially water vapor impermeable" means that substantially gaseous water can not enter the interior of the container from the outside through the material of the container, but in preferred embodiments, gaseous water may pass through the container material from inside to outside, that is, the water from However, in alternative embodiments, essentially no gaseous water, either from outside to inside, or from inside to outside, can pass through the container material.

Depending on how dense the vapor barrier of the material is, some essentially anhydrous blood plasma samples must be stored in particularly dry rooms. In order to save this again somewhat increased storage costs, in preferred embodiments, the material is selected so that the passage of gaseous water is completely excluded by this material at least from the outside inwards. Advantageously, plastic bags are used as containers for the blood plasma samples. Appropriate Plastic materials with barrier properties or with barrier coatings on the inside or outside are known. When using plastic materials that do not or not sufficiently have such water vapor barrier properties, the use of secondary packaging, e.g. As aluminum, be advantageous.

For the purposes of this application, plasma end volume is the volume of the plasma resolubilized for administration to a patient. In preferred embodiments, the maximum internal volume of the container is at least 5% greater than the final plasma volume. As a result, in particular during filling and removal, improved handling of the bag is achieved when used in accordance with the present invention.

It is understood that all materials used and measures taken in the use must be chosen or performed so that the sterility of the blood plasma, regardless of its consistency, always maintained.

To prepare a blood plasma bag for use according to the invention in volume replacement or volume exchange therapy, the blood plasma taken from a donor / patient under sterile conditions is introduced into a sterile, substantially water vapor impermeable container, then the blood is substantially completely withdrawn from the blood plasma, after which it is sealed the container waterproof and sterile and then preserves the substantially anhydrous blood plasma in the container and then opens the container only at a desired or required time again, brings sterile water into the container to the substantially anhydrous blood plasma to obtain liquid Dissolve blood plasma in physiological concentration. This reconstituted liquid autologous blood plasma is then ready for use in volume replacement or volume exchange therapy after ensuring that all components of the dry plasma are completely dissolved.

The whole blood withdrawn to obtain the blood plasma is preferably centrifuged and the plasma obtained is squeezed into a transfer bag. Alternatively, the plasma is recovered by a machine apheresis procedure.

Ideally, the freshly recovered plasma is directly dried or lyophilized within 24 hours. Alternatively, the plasma may also be flash frozen within 24 hours and then stored at at least -30 ° C before being added to it

Thawing time and further processed according to the invention. In pool plasma or other In any case, plasma donations that are not self-plasma will be released for use at the earliest after four months of quarantine storage, and if transfusion-relevant infection markers are still undetectable in a later blood sample from the donor / donor. In the case of self-plasma, on the other hand, compliance with this quarantine phase and the subsequent examination of the donor for transfusion-relevant infection markers can advantageously be dispensed with. After expiry of the quarantine storage, the plasma can additionally be subjected to virus inactivation and depletion, as is known in the prior art (eg SD plasma, solvent / detergent method), which of course is omitted in the case of the intrinsic plasma. The transfusion takes place exclusively blood group specific, so ABO equal, which automatically results in the self-plasma.

In an alternative embodiment, the blood plasma is not introduced immediately after removal into the substantially water-vapor-impermeable container, but first frozen and then weighed into this container in a frozen state. In another alternative embodiment, the blood plasma is dried before being weighed into the aforementioned container in this substantially anhydrous state.

The removal of the water from the blood plasma introduced into the substantially water vapor-impermeable container takes place at least in part by lyophilization and / or by vacuum extraction. Preferably, the removal of the water is complete by lyophilization and / or vacuum extraction.

An advantage of the plasma preparations prepared according to the invention is that the dry plasma can be brought to body temperature faster and with less technical effort than conventional frozen plasma that can be thawed and warmed only with specially approved equipment to ensure that no overheating of the plasma to more than + 42 ° C.

As already mentioned above, all process steps must be carried out with the proviso that the sterility of the blood plasma is not impaired.

Further features and characteristics of the invention as well as advantages that result from individual features or combinations of features or combinations of feature groups will become apparent from the specific embodiment of the present invention described below by way of example. In considering these exemplary embodiments, it should be noted that not all embodiments of the present invention are all implemented in the exemplary embodiment Features must be realized in order to implement the present invention and to take advantage of its benefits.

Example 1 Preparation of a plasma preparation (525 ml final plasma volume including stabilizer)

The following procedure is based both on the current method of blood and plasma collection as well as the goal of preparing in practice, round volumes of distilled water. use.

For the inventive use of blood plasma for long-term storage and the subsequent use in the volume replacement or volume exchange therapy, in this example a donor is taken in a conventional manner two whole blood single donations (2 times 450 ml), wherein the blood 106 ml CPD solution (stabilizer solution) be added for coagulation inhibition, an amount corresponding to 20 vol .-% based on the final volume of the pure plasma.

Subsequently, the staggered with stabilizer solution is centrifuged, whereby all cellular components are separated from the plasma. Thereafter, the cell-free plasma is squeezed into a transfer bag and subjected within 24 h of the further processing described below by lyophilization. If no further processing is required, the plasma is frozen within 24 hours, stored at -30 ° C until further processing and further processed within 12 months at the latest.

For further processing, 525 ml of plasma are placed in a sterile closable and substantially water vapor impermeable storage bag having a capacity of about 550 ml, i. H. about 5% more than the plasma volume. Subsequently, the substantially complete removal of water (500 ml) by lyophilization on or in a freeze dryer.

The completeness of the water removal can be checked at given times on the basis of the weight determination of the dried sample. In the present case, the blood plasma preparation is substantially free of water when only about 38 g of dry matter (plasma lyophilisate) are present in the bag. If this state is reached, the sterile closing of the bag and the subsequent storage takes place at room temperature. Depending on For intended use, the bag is marked with the manufacturer, the date of manufacture and the name, date of birth and, if applicable, the passport photograph of the donor.

If necessary, recourse is had to the blood plasma stock prepared in this way by connecting the storage bag to a sterile container containing approximately 500 ml of distilled water. This volume is then introduced into the dry plasma container for dissolving the lyophilisate under visual control and is then ready for transfusion to the donor very quickly for the purpose of volume replacement or volume exchange therapy.

When the plasma is removed as a machine apheresis plasma, an ACD stabilizer solution (aqua citrate-dextrose solution) in an amount of 18.5% by volume based on the final volume of the pure plasma can also be used. In this case, directly 525 ml of plasma including 18.5 vol .-% ACD stabilizer are obtained. Further processing takes place as described for the individual donation.

The ratio of the weight of the dry substance (plasma lyophilisate) to the removed water is determined as follows:

Plasma has a dry matter content of about 80 g / l at a density of 1.0269 kg / l and a water content of 945 g / l. The stabilizer (CPD solution or ACD solution) has a

Dry matter content of about 56 g / l and a water content of about 990 g / l.

Accordingly, the addition of 20 vol .-% stabilizer solution to the plasma of

Total water content increased to about 954 g / l and reduced the dry matter content to about 75 g / l. 525 ml of plasma including 20% by volume of stabilizer solution give about 500 g or ml of water during lyophilization and about 37.5 g of dry substance remain.

If this preparation is then filled up with 500 ml of distilled water, the result is again

Plasma volume of 525 ml including stabilizer solution.

Claims

1. Use of autologous blood plasma (self-plasma) of a patient in substantially anhydrous form in a sterile sealed, substantially wasserdampfundurchlässigen container whose inner volume may include at least one of the plasma end volume corresponding volume, for long-term storage and subsequent production of liquid autologous blood plasma (self-plasma) for the volume replacement or volume exchange therapy.

2. Use according to claim 1, characterized in that the maximum internal volume of the container is at least 5% greater than the plasma end volume.

3. Use according to one of claims 1 and 2, characterized in that a container is used whose internal volume is variable, wherein the container consists at least partially of flexible material.

4. Use according to one of claims 1 to 3, characterized in that the blood plasma is lyophilized and / or vacuum-extracted.

5. Use according to one of claims 1 to 4, characterized in that the storage time is longer than 1 year, wherein the storage time is preferably at least 5 years, more preferably at least 10 years.

6. Use according to any one of claims 1 to 5, characterized in that the blood plasma is mixed with a stabilizer solution, preferably admixed with a citrate-containing stabilizer solution.

7. Use according to one of claims 1 to 6, characterized in that the container is substantially impermeable to water vapor only in the direction from outside to inside and is permeable to water vapor from the inside to the outside.

8. Use according to any one of claims 1 to 7, wherein: a) sterile autologous blood plasma (own plasma) of a patient is introduced into a sterile, substantially water vapor impermeable container, b) the autologous blood plasma substantially completely withdraws the water, c) the container waterproof sterile closes, - -

d) storing the substantially anhydrous autologous blood plasma in the receptacle; e) reopening the receptacle at the desired time; and f) introducing sterile water into the receptacle for dissolving the substantially anhydrous autologous blood plasma to give liquid autologous blood plasma in physiological concentration.