WO2000059551A1

WO2000059551A1 - Photodynamic inactivation of viruses in biological fluids

Info

Publication number: WO2000059551A1
Application number: PCT/EP2000/002845
Authority: WO
Inventors: Gerhard Saalmann; Peter Saalmann; Norbert H. Brockmeyer; Klaus Hoffmann; Peter Altmeyer; Darius Alamouti
Original assignee: Gerhard Saalmann; Peter Saalmann
Priority date: 1999-04-01
Filing date: 2000-03-31
Publication date: 2000-10-12
Also published as: DE19914850A1; AU3816300A

Abstract

A device and a process for the inactivation of viruses in biological fluids, especially for the inactivation of viruses -- especially HIV -- in blood and blood products in which phenothiazine dyes, especially toluidine blue or methylene blue, are added to the biological fluid; a single-part or multipart pumping and transport device is used to collect the biological fluid from a container or circulation, the biological fluid passing through a system of conduits and returning to the container or to the circulation. A separating device serves to separate the biological fluid into a liquid and a corpuscular phase. A light source emits electromagnetic radiation of the visible spectrum onto a light-permeable segment of the conduit system.

Description

PHOTODYNAMIC INACΗVATION OF VIRUSES IN BIOLOGICAL FLUIDS

FIELD OF THE INVENTION

The invention concerns a device and a process for the inactivation of viruses in biological fluids, especially for the inactivation of viruses - especially HIN - in blood and blood products in which photosensitizers, preferably phenothiazine dyes, especially toluidine blue or methylene blue, are added to the biological fluid.

DESCRIPTION OF THE RELATED ART

European patent 0 491 757 Bl shows a process for the inactivation of viruses in blood in which the solutions or suspensions to be treated are mixed with phenothiazine dyes and subsequently bombarded with electromagnetic radiation. The phenothiazine dyes are used in a concentration of 0.1 to 2 M, and the irradiation takes place directly in transparent containers such as blood pouches which are used for collecting and storing blood. Phenothiazine dyes react with the membrane structures of sheathed viruses or with the viral DNA and RNA and damage them under the influence of visible light.

EP 0 471 794 Bl shows the use of xanthene or thiazine dyes for the production of a pharmaceutical agent for the selective inactivation of the HIV in vivo or in vitro without significant toxicity for cells or the patient. The xanthene or thiazine dyes were tested on peripheral blood mononuclear cells. The treated cells were irradiated after treatment with dye. Testing on whole blood or plasma was not reported.

US Patent No. 4,737,140 to Lee et al. shows an irradiation chamber for extracorporeal phototherapy of blood. The apparatus collects and separates blood on a continuous basis as it is withdrawn from a patient and returns unwanted portions to the patient. The remainder is passed through an irradiation chamber where it is subjected to phototherapy. The patent is directed to cancer treatment. The use of dyes and the treatment of viral infections are not shown. Infection by the human immunodeficiency virus (HIV) induces directly or indirectly an immunity weakness which is most clearly reflected in the loss of the CD4-positive T-lymphocytes. Other cells which carry CD4 receptors are the monocytes, macrophages, dendritic cells, microglia and nerve cells. The results of this immunity weakness are HIV disease and AIDS which are characterized especially by the occurrence of opportunistic infectious diseases, dermatoses and tumors. According to the results of a recent study, therapy is now administered not only based on the CD4 cell count/μl or the appearance of HIV-associated diseases but also based on the virus load (HIV-RNA copies/ml). It is known that patients with a high virus load display a progressing course of the disease. In the meanwhile, a large number of preparations have become available for therapy. The goal of the therapy must be to lower the virus load by at least two logio steps or to bring it below the limits of detection.

Possible therapy failures or development of resistance to viral enzymes, e.g. reverse transcriptase or viral protease represent problems that could lead to ineffectiveness of the therapy. Here the use of thiazine dyes as pharmaceutical agents has aroused new hopes. In light of the well known low toxicity of certain thiazine dyes (methylene blue and toluidine blue have long been used among others as antidotes for carbon monoxide poisoning), the use of these dyes as drugs against HIV appears especially advantageous. HIV-1 was found actually to be especially susceptible to inactivation by these substances. At the same time the xanthene or thiazine dyes as drugs or as constituents of an effective virus reduction system for the selective inactivation of HIV have heretofore failed to achieve any noteworthy success. This is due to the possibility of damage to lymphocytes.

Against this background, the invention has as an objective, developing a device and a process for reducing the viral load of biological fluids - especially the blood of a patient ~ which achieves a significant reduction in the viral load of the biological fluid without major functional impairments of the lymphocytes of the blood. By the term "viral load" as used herein, is meant the number of viable active virus copies in the blood in terms of copies per unit volume.

These and other objects are achieved by the present invention.

SUMMARY OF THE INVENTION

The present invention pertains to a device or for the inactivation of viruses in biological fluids, especially for the inactivation of viruses — especially HIV - in blood and blood products, in which photosensitizers, preferably phenothiazine dyes, especially toluidine blue or methylene blue, are added to the biological fluid with: a single-part or multi-part pump and transport device for collecting the biological fluid from a blood circulation, for passing the biological fluid through a conduit system and for returning the biological fluid to the blood circulation, a separating device for separating the biological fluid into different liquid phase and/or liquid and solid phases, especially for separation into blood plasma and corpuscular elements; in which the conduit system and/or a container connected to the conduit system are designed to be transparent at least in some segments; and with a light source for emitting electromagnetic radiation, especially of the visible spectrum, onto the light- permeable conduit system segment or the light-permeable container.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 is a schematic drawing of the device of the present invention.

Fig. 2 is a schematic drawing of an alternative embodiment of the device of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

According to the invention there is provided a single-part or multi-part transport device for collecting the biological fluid from the blood circulation, for passing the biological fluid through a conduit system, and for returning the biological fluid to the blood circulation. The blood is taken from the blood circulation of the patient in order to be directly bombarded with visible light in the transparent conduit system or container or in the separating device. The blood may be any blood that can be infected with a virus, although the present invention will be most useful with mammals. Typical mammals with which the present invention is useful include humans, dogs, cats, cows, pigs, monkeys, apes, sheep, goats, horses, rodents, mice, rats, and rabbits.

In order to avoid the undesired destruction of proteins at this time the invention makes use of a separating device for dividing the blood up into different liquid phases, especially plasma and corpuscular components, where then only the plasma phase is irradiated with visible light. After this all phases of the blood are returned together or separately to the blood circulation. In addition to direct treatment of a patient, the process is also suitable for the treatment, from containers of any kind, of biological fluids that can be divided into phases and can be brought together again after the application of the "phototherapy".

In different forms of therapy, the blood of a patient can be irradiated with UV radiation (see U.S. Patent 4,737,140). However, the present invention is a vast improvement over such older technologies.

The mode of action of photopheresis is largely unknown. It is possible that photopheresis leads to a modification of the defense system (immunomodulation). Malignant cells or cells which are responsible for the production of autoantibodies are possibly recognized as foreign by the immune system after the treatment and attacked. UV irradiation is a therapy procedure introduced long ago in which the skin is irradiated with ultraviolet radiation (UV-A or UV-B). The effect can be improved with the aid of locally or systemically administered photosensitizers in combination with UV-A radiation (PUVA therapy). A further development of this therapeutic process is extracorporeal PUVA therapy (ECP). In this process it is no longer the skin but rather directly the lymphocytes in the blood which are irradiated. In this case venous blood is collected through a cannula in the same manner as for blood donation. The blood is then sent into the extracorporeal system. The white blood corpuscles are separated from the red blood corpuscles and the blood platelets by a centrifugation or other separation step. The red blood corpuscles and blood platelets are preferably immediately reinfused back into the patient.

Despite the fact that general photopheresis process has been used experimentally in some university clinics, an adaptation of this process as set forth in the present invention for a new type of photopheresis therapy for reducing the virus load of the blood utilizing a centrifuge, irradiation with visible light and phenothiazine dyes offers a much improved method of fighting viral infections.

According to a preferred variant, the light source and the separating device (centrifuge or filtration unit) are combined for design simplification into a single structural unit.

Preferably, the centrifuge or the filtration unit has light-permeable walls, at least in segments, and a light source is aligned in such a way that the electromagnetic radiation strikes at least one of the liquid phases through the light-permeable walls of the centrifuge. In this way a separate irradiation unit can be omitted.

The radiation unit can be used to produce desired wavelengths of electromagnetic radiation. This radiation should preferably be in the range of 380 to 780 nm to correspond to the absorption spectrum of different thiazine dyes. The spectrum of radiation of the light source more preferably includes the wavelengths from 550 to 700 nm. The emitted radiation spectrum of the light source should have its highest intensity approximately in the range of the absorption maximum of the dyes used in each case, and this will vary with the particular dyes used.

According to one variant of the invention, a sufficiently long irradiation time is achieved by widening the light-permeable conduit system in the region of the light incidence peφendicular to the light incidence in such a way that, due to the enlargement of the cross section of the conduit system related to this, the flow velocity in the conduit system of the biological fluid in it is reduced. Thus the cross- sectional area of the conduit is increased, to decrease the linear velocity of the fluid. However, this increase in the cross-sectional area must be balanced. If the increase is too great, the radiation may not reach all of the fluid in the conduit. Therefore, the widening of the conduit must be tempered. Alternatively, a second transparent area may be introduced to allow for greater phototherapy. In such a case, the same light source may be used, or an additional light source may be added.

For simpler manipulation in another variant a vein adapter or sensor can be connected preferably to the reservoir interface. A temperature regulating device wired into the conduit system for regulating the temperature of the biological fluid makes the necessary cooling of the blood possible.

The device preferably includes a metering device connected into the conduit system for releasing the dyes into the biological fluid.

The invention also creates a new application for known drugs, that being the application of phenothiazine dyes, especially toluidine blue or methylene blue, for the production of a drug for the inactivation of viruses in a therapy process of the type explained below.

The invention also creates a process for inactivation of viruses in biological fluids, especially for the inactivation of viruses — especially HIV — in blood and blood products, which involves adding photosensitizers to the biological fluid, preferably phenothiazine dyes, especially preferably toluidine blue or methylene blue, with the following steps: the biological fluid is separated by a separating device into different liquid phases and/or liquid and solid phases, especially into blood plasma and corpuscular elements, one of the liquid phases (i.e. plasma) is bombarded with electromagnetic radiation, especially of the visible spectrum, until part of the viruses present in the blood has been essentially destroyed or deactivated, and the liquid and/or solid phases are brought together again after irradiation and/or mixed with each other again. The irradiation is performed on the blood fraction which is free, or substantially free, of red corpuscles to prevent destruction of those cells. Finally the invention is also especially suited for devising a process for the inactivation of viruses in biological fluids, especially for the inactivation of viruses - especially HIV ~ in blood and blood products. Photosensitizers, preferably phenothiazine dyes, especially preferably toluidine blue or methylene blue, added to the biological fluid. The blood is taken from a blood circulation and, before or after being taken from the circulation, the phenothiazine dyes are added to the blood. The venous blood is separated in the separating device into a first phase with the main component plasma and a second phase with the main component coφuscular elements. The first (plasma) phase, with a small - especially scarcely measurable - content of white blood coφuscles, is irradiated with electromagnetic radiation, especially of the visible spectrum, until the viruses present in this blood phase have been largely destroyed or deactivated by the electromagnetic radiation. The two liquid phases and/or liquid and solid phases are returned to the circulation after irradiation.

The invention further teaches a use of photosentisizers, especially phenothiazine dyes for producing a drug for inactivating virusses in according with a method of one of the preceeding claims.

Other advantageous variants of the invention will be apparent to one skilled in the art.

The following the invention is described in more detail with reference to the drawings.

Figure 1 illustrates a device 2 for inactivation of viruses in biological fluids, especially for the inactivation of viruses — especially HIV — in blood and blood products. As used herein, the term "blood" refers to whole blood, and the term "blood products" refers to any portion or portions of whole blood, including, but not limited to, erythrocytes, leukocytes, plasma, or any other portion or fraction of whole blood.

The blood is passed from a container 4 (or directly from the blood circulation of a patient) through a valve 6 in a conduit 8 of a conduit system. A pump 10 is connected to the conduit 6 with which the blood can be pumped in the conduit 8 in both directions. Behind the pump 8 a conduit branch 12 with a valve 6 makes it possible to add an anticoagulant from a feeder device 14 designed for this task. The feeder device 14 or another (not shown here) feeder device may, if necessary, also be utilized for the direct addition of the thiazine dyes to the blood of the patient.

From the pump the pumped-in blood is fed via a third valve 6c and a conduit branch 16 into a centrifuge or a filtration device 18 (e.g., a PRISMA^™ CFR microfiltration unit trademark of HOSPAL Medizintechnik GmbH, Brettergartenstr. 16 90477 Nuremberg, Germany). The centrifuge or the filtration device 18 makes it possible to separate the fed-in blood into two liquid phases and/or liquid and solid phases. One of these phases (with a high content of red blood coφuscles) is sent via the branch line 20 with a valve 6d to a return pouch 22 or directly back to the container 4 or a patient's circulation. The other phase is sent via a branch line 24 with a valve 6d to a plasma container 26. From the plasma container 26 the blood plasma is then sent through an irradiation cycle 28 with inflow line 30, pump 32, irradiation unit 34 with (not shown) cooling and return line 36.

Finally the blood is returned from the plasma container 26 via the centrifuge or filtration device 18 and the return pouch 22 and a return branch line 38 with valve 6f as well as the pump 10 and conduit 8 with the valves in the appropriate position to container 4 or to the blood circulation of a patient.

The example in Figure 2 differs from this example of embodiment essentially in the fact that the blood is passed from the pump 10 directly to a centrifuge/irradiation unit combination 40 where the lamp segment 42 emits light onto the centrifuge part 44 with walls with light-permeable segments.

An exemplary treatment with the above-described device follows:

For a quantity of 500 ml plasma one should use between 0.005 and 0.025 ml, preferably 0.010 to 0.020 ml, most preferably approximately 0.015 ml of 1% sensitizer. The photosensitizer is most conveniently in an aqueous solution of between 0.1 and 10%, preferably 0.5 to 5%, most preferably about 1% sensitizer. Any concentration may be used, however higher concentrations are difficult to work with as small errors in measurement dramatically affect dosage. Similarly, low concentrations add unnecessary volume to the treatment dosage. Of course, the volume used is dependent on the sensitizer concentration. The photosensitizer may be toluidine blue or methylene blue (MB). The radiation source should operate with visible light (380-780 nm). Fluorescent tubes (e.g. Phillips TL-M 115 W/3 RS) or quartz lamps with which the plasma is irradiated for 1 h with 50,000 lux are suitable. The temperature regulating system maintains a temperature of a maximum of 30°C.

The HIV burden is reduced by the treatment. An additional advantage of this therapy is the fact that other viruses can also be inactivated (heφes simplex, heφes zoster, influenza, vesicular stomatitis, Semliki forest, adeno, polio, cytomegalo, etc.). The leukocytes/lymphocytes, conversely, are not or only slightly influenced by it. Coagulation proteins and blood constituents remain intact.

At first venous blood is taken from the patient. The white blood coφuscles are separated from the red blood coφuscles and the blood platelets by a centrifugation step (ca. 4800 φm + 5%). The red blood coφuscles and the blood platelets are immediately reinfused back into the patient. These cycles are repeated until one has collected max. 15% of the total blood volume (therefore max. ca. 600 ml) of plasma including solitary leukocytes. Because of the above-mentioned cycles, a high blood volume throughput and accordingly a high HIV virus concentration in the collected plasma is guaranteed. During the process a conventional coagulation-inhibiting drug such as heparin is added to the collected venous blood so that the blood does not clot and clog the machine when flowing through the system of hoses.

After the collection of the plasma under sterile conditions, the corresponding quantity of a sterile 50 μl solution of MB in water for injection is added. The final concentration of MB is 1 μM. This photoactive substance has a strong affinity for the surface structures of viruses and viral nucleic acids and can therefore bind to them. When exposed to light it absorbs energy and transfers it to the molecules to which it is bound. Ultimately this results in the denaturing of the components of the virus shell or breaks in the strands of the viral HIV RNA. Highly reactive oxygen radicals are also involved in this process. It has also been demonstrated that the dyes interfere with transcription and translation (of DNA and RNA) in darkness as well as in a weak or a strong light. Preferably the plasma is irradiated for 1 hour by the light source (50,000 lux). However the time necessary for destruction or inactivation of viruses will depend on many factors including the particular virus, the dye concentration, the particular dye used, the temperature, the light intensity, and the wavelength used, among others. This process is temperature dependent, i.e. the operating temperature must be between 20 and 30°C so that any proteins still present are not destroyed. After the irradiation time the plasma is reinfused back into the patient. The treatment (collection and irradiation process) could be performed initially several times daily. The therapy cycle is repeated, depending on the virus load of the patient, from initially, e.g., five times weekly to once per month.

The invention is now illustrated by non-limiting, representative examples.

Example 1

This experiment shows the virus inactivation properties of methylene blue and light in plasmas from four different patients.

TABLE 1

Irradiation time/minutes HIV copies/ml without HIV copies/ml with methylene 3lue Methylene blue

0 814 Below detection limits

60 743 Below detection limits

60 10751 Below detection limits

60 9275 Below detection limits

60 3944 Below detection limits With the aid of the AMPLICOR HIV-1 Monitor Test (version 1.5) from Roche the

HIV RNA in the treated plasma could be detected quantitatively. HIV was found to be especially susceptible to the photo-inactivation step. The intensity of the total inactivation for HI viruses in the presence of methylene blue, however, depends on the duration of the exposure. In order to investigate what exposure times are sufficient for photoinactivation of HIV, 10 test tubes containing plasma were exposed at 27°C for different times.

TABLE 2

Number Irradiation time (min) Virus Copies/ml

1 negative control test -3

2 Control 790,899

3 0 868,189

4 5 583,374

5 10 154,375

6 15 22,593

7 20 19,943

8 30 2,495

9 40 414

10 50 89

11 60 below limit of detection

Example 2

The second example illustrates the effect of the photodynamic therapy on the lymphocyte population. First at lymphocyte differential count of a healthy subject is analyzed. From the reference range, it can be seen that this lymphocyte differential count is well within the normal range. A conventional photodynamic therapy was performed with this patient's plasma (including lymphocytes):

TABLE 3

Sample Annexin Propidiurr l iodide + Annexin

24 hr. culture without MB without 7.9% 1.32% light

24 hr. culture without MB with light 10% 1.82%

24 hr. culture with MB with only 13.6% 7%

50% of usual light intensity

24 hr. culture with MB with light 16.4% 27.5%

MB = methylene blue

Example 3

With the Annexin V-FITC Apoptosis Detection Kit from Pharmingen and the DNA-binding dye propidium iodide we show the increase of the programmed cell death (known as apoptosis) and of the cells shortly before the necrosis. It can be seen that the measurements of the apoptosis and necrosis increase from normal (here 1.32%) up to 27.5%o. This is proof that methylene blue not only harms the viruses but also the lymphocytes, a phenomenon that the present invention avoids. Example 4 - Patient A; age 48

All values were checked 3 times and averaged.

On Day 1, the patient received orally 10 mg/kg body weight methylene blue, i.e.

845 mg of methylene blue, for the first time. After 3-4 hours the blood concentration of 0.5 μmol/1 methylene blue had been exceeded. After 6 h the half-life was already reached.

At 845 mg methylene blue taken orally no abnormalities appeared in the differential lymphocyte count (after 24 h), see Table 4 Day 1 to Day 2. At this time a reduction in the virus load from 655 copies/ml to below the detection limit was noted.

From Day 8 until Day 9 the patient received an initial dose of 10 mg/kg body weight at noon and then after 6 hours 5 mg/kg body weight, and on the next morning again 5 mg/kg body weight. After the first dose of MB on Day 9, the patient received an additional extracoφoreal irradiation of ca. 1000 ml plasma with visible light. At that time a reduction in the virus load by MB on Day 8 from 190 copies/ml to below the detection limit was noted and the next morning with only a single dose of MB with extracoφoreal irradiation the virus load dropped from 333 to 100 copies/ml. If the therapy was terminated, the virus load increased again. The same patient was hospitalized again on Day 24 but this time for a venous thrombosis of the lower leg. He was put on Marcumar. We took advantage of this opportunity and continued the study. From Day 25 to Day 30, he received 10 mg/kg body weight every day in the morning and then 5 mg/kg body weight every 5 hours. The virus load dropped from 167 to 106/ml. One day after termination the virus load had risen again. During the entire treatment time no abnormalities were seen in the differential lymphocyte count, clinical chemistry, electrolytes, glucose, rheumatoid serology, thyroid gland hormones, coagulation and hematology. EKG and chest x-ray of Day 28 were normal.

TABLE 4 - Differential leukocyte count - Patient A

Example 5 - Patient B, Age 40

On Day 1 the patient received 10 mg/kg body weight methylene blue orally for the first time. After 1 h the blood concentration of 0.5 μmol/1 methylene blue had been exceeded. After 7 h the half-life was already reached.

At 780 mg methylene blue taken orally no abnormalities appeared in the differential lymphocyte count (after 24 h), see Table 5, Day 1 to Day 2. At this time a reduction in the virus load from 666,000 to 612,000 copies/ml was noted. From Day 8 until Day 9, the patient received an initial dose of 10 mg/kg body weight at noon and then after 6 hours 5 mg/kg body weight and the next morning again 5 mg/kg body weight. After the second dose of MB on Day 8, the patient received an additional extracoφoreal irradiation of ca. 1000 ml plasma with visible light. At that time a reduction in the virus load by 2 x administration of MB on Day 8, from 582,000 to 320,000 copies/ml was noted and after additional extracoφoreal irradiation of 1000 ml plasma the virus load dropped further from 320,000 to 178,000 copies/ml. If the therapy is terminated, the virus load increases again. The plasma of the same patient was irradiated again on the next day after taking MB. The virus load drops from 761,000 to 253,000 copies/ml. Six hours after completion the virus load had increased again. During the entire treatment time no abnormalities were seen in the differential lymphocyte count, clinical chemistry, electrolytes, glucose, rheumatoid serology, thyroid hormones, coagulation and hematology.

TABLE 5 - Differential leukocyte count - Patient B

1*1 ks)

.O

Example 6 - Patient C, Age 46

From Day 1 until Day 8, the patient received an initial dose of 10 mg/kg body weight, then every 5-6 hours 5 mg/kg body weight. Also no abnormalities were seen in the differential lymphocyte count, clinical chemistry, electrolytes, glucose, rheumatoid serology, thyroid hormones, coagulation and hematology. Abdominal ultrasonography (on Day 8) revealed a long known minor hepatomegaly and distinct splenomegaly as well as steatosis hepatitis. Chest x-ray and EKG (on Day 8) were normal.

TABLE 6 - Differential leukocyte count - Patient C

The inversely proportional relationship between MB concentration in the blood and virus load/ml in the blood is clear, and the efficacy is obvious. This therapy approach shows that the results achieved in vitro correlate with the in vivo results. If the range of 0.5 μmol/1 methylene blue is reached the virus copies are reduced without presently demonstrable objective or subjective side effects for the patients including the extracorporeal irradiation. Although the necessary virostatic plasma levels were reached for only four days, the results were clear. The purpose of the extracorporeal irradiation in the intensive initial treatment is to reduce the virus load very quickly and then to implement further oral antiretroviral therapy. This also applies to patients who cannot undergo oral therapy because of kidney and liver disease or who would rather not be in a condition of permanently blue- stained urine and feces.

Example 8 - Monkey

Other mammals may be treated in accordance with the method of the invention.

Venous blood is taken from a monkey infected with a simian analog of HIN.

The blood is separated using a centrifuge, and the platelets and red blood cells

are returned immediately to the monkey. This is repeated until about 10% of

the total blood volume is gathered as plasma. To the plasma is added

methylene blue, at a rate of about l μl/10-15 ml plasma. The plasma is

subjected to phototherapy for one hour while circulating through a temperature

controlling device, to maintain a temperature between 20 and 30°C. The

plasma is then infused back into the monkey. The plama may be tested to

confirm a significant reduction in virus load. The treatment can be performed

several times per week, or even several times per day when necessary.

Claims

Claims:

1. Device for the inactivation of viruses in biological fluids which contains photosensitizers, comprising: a pump, collecting means for collecting the biological fluid from a source, and a conduit system operatively connected to said collecting means and pump for returning the biological fluid to the source, the conduit system having at least one transparent segment, a separating device for separating the biological fluid into a first liquid phase and a second phase, and a light source for emitting electromagnetic radiation onto the transparent conduit system segment.

2. The device of claim 1 wherein the photosensitizers comprise phenothiazine dyes.

3. The device of claim 2 wherein the photosentizers are selected from the group consisting of toluidine blue and methylene blue.

4. The device of claim 1 wherein the source is a human circulatory system.

5. The device of claim 1 wherein the conduit system includes a container and the at least one transparent segment forms a part of the container.

6. The device of claim 1 wherein the light source emits electromagnetic radiation in the visible spectrum.

7. The device of claim 3 wherein the source is a human circulatory system, the conduit system includes a container, the at least one transparent segment forms a part of the container, and the light source emits electromagnetic radiation in the visible spectrum.

8. The device of claim 3 wherein the conduit system includes a container and the at least one transparent segments forms a part of the container.

9. The device of claim 8, wherein the separating device is one of a centrifuge and a filtration unit, the biological fluid is blood, the first liquid phase thereof contains plasma and the second phase thereof contains corpuscular elements.

10. The device of claim 8 wherein the light source and the separating device are combined into a single structural unit.

11. The device of claim 8 wherein the transparent conduit segment forms a part of the separating device and the light source is aligned such that its electromagnetic radiation passes through the transparent segment onto at least one liquid phase.

12. The device of claim 8 wherein the emission spectrum of the light source encompasses the wavelengths from about 550 to 700 nm.

13. The device of claim 8 wherein the emission spectrum of the light source has its highest intensity approximately in the range of the absorption maximum of the photosensitizer.

14. The device of claim 11 wherein the transparent segment has a conduit thickness selected such that the electromagnetic radiation essentially totally penetrates the biological fluid.

15. The device of claim 3 wherein the cross-sectional area of the transparent segment perpendicular to the path of incidence of the light is greater in the region of the incidence of the light or radiation than in other areas, such that the flow velocity of the biological fluid is reduced in the transparent segment.

16. The device of claim 8 further including a vein adapter.

17. The device of claim 8 further including a temperature regulating device connected to the conduit system.

18. The device of claim 8 further including a metering device connected to the conduit system for releasing the photosensitizer into the biological fluid.

19. The device of claim 4, wherein the source is the human circulatory system which is adapted to be connected to a vein of the human.

20. A process for the inactivation of viruses in biological fluids in which photosensitizers are added to the biological fluid, comprising the following steps:

separating the biological fluid into a first, virus containing liquid phase and a second phase, irradiating the first liquid phase with electromagnetic radiation, sufficient to inactivate or destroy at least a portion of the viruses present in the blood, and recombining the two phases.

21. The process of claim 20 wherein the photosensitizers are selected from toluidine blue and methylene blue.

22. The process of claim 20 wherein the biological fluid is blood, and the first liquid phase includes plasma and the second phase includes corpuscular elements.

23. The process of claims 22 wherein the first phase has an elevated content of white blood corpuscles and is irradiated with electromagnetic radiation of the visible spectrum.

24. A process for inactivation of viruses in blood comprising adding a photosensitizers selected from toluidine blue and methylene blue to the blood, withdrawing blood from a living organism, and separating the blood into a first plasma phase and a second corpuscular phase, irradiating the first plasma phase with electromagnetic radiation of the visible spectrum, for a time and at an intensity sufficient to destroy or inactivate substantially all of the viruses present in the first plasma phase, and returning the two phases to the organism.

25. A process for treating a viral infection in a mammal comprising withdrawing blood from the mammal, adding a photosensitizer to the blood, separating the blood into a first liquid phase and a second phase, irradiating the first phases, and returning the irradiated phase into the mammal.

26. The process of claim 25 wherein the blood is separated into a first liquid phase and a second solid phase comprising corpuscular elements.

27. The process of claim 26 wherein the irradiated phase is the first liquid phase.

28. The process of claim 27 including the step of returning the second, sqlid phase into the mammal.

29. The process of claim 28 wherein the mammal is a human.

30. The process of claim 29 wherein the viral infection is caused by a human immunodeficiency virus (HIV).

31. The process of claim 30 wherein the photosensitizer is a phenothiazine selected from the group of toluidine blue and methylene blue.

32. The process of claim 31 wherein the first liquid phase is irradiated with visible light.

33. A process for the inactivation of viruses in human plasma which contains a photosensitizer selected from the group of toluidine blue and methylene blue, which comprise irradiating to the plasma, and thereby producing a treated plasma;

34. The treated plasma produced by the process of claim 33.

35. The process of claim 33, wherein the treated plasma is fed into a vein of the human. WO 00/59551 . PCT/EPOO/02845

36. The process of claim 33 wherein the plasma, which contains a human immunodeficiency virus, is exposed to visible light in the range of about 550 to 700 nanometers.

37. The process of claim 36 wherein the plasma is mixed with a phase comprising corpuscular elements, thereby obtaining reconstituted blood.

38. A constituted treated blood produced by the process of claim 37.

39. The process of claim 37 comprising further feeding a second phase comprising corpuscular elements separately from the treated plasma to the mammal.

40. The process of claim 35 or 39 repeated at least twice in a 24 hour period.

41. The method of claim 37 wherein the reconstituted blood is fed into a container.

42. An extracorporeal reconstituted blood of plasma and corpuscular elements, which plasma contains a photosensitizer, which is radiated with electromagnetic radiation in the visible spectrum, and substantially HIV-aviral, the reconstituted blood being adapted to be venously fed into a human.

43. The reconstituted blood of claim 42, which is under physiological conditions.

44. Use of photosentisizers, especially phenothiazine dyes for producing a drug for inactivating virusses in according with a method of one of the preceeding claims.