AU608545B2

AU608545B2 - An improved method

Info

Publication number: AU608545B2
Application number: AU80548/87A
Authority: AU
Inventors: Horst Dr. Kief
Original assignee: HORST KIEF DR
Current assignee: HORST KIEF DR
Priority date: 1986-10-30
Filing date: 1987-10-29
Publication date: 1991-04-11
Anticipated expiration: 2007-10-29
Also published as: EP0265548B1; EP0265548A1; AU8054887A; ZA878117B; ES2017618B3; ATE56363T1; GR3001178T3; JPS63188391A; DE3674213D1

Abstract

1. Process for the preparation of substances (suspensions) which have been sterilized or whose virulence has been attenuated, by extra corporeal means, where the original substances which have been removed from a living, diseased individual are subjected to an oxidation by means of ozone, oxygen and/or ionising radiation, characterized in that the original substances are fractionated.

Description

Iillaili i :i lillil 1 iII jI l lji 111~ I.T11; 11111 IA 'V 5263-P1 JGS:GS 3961T/9

AUSTRALIA

PATENTS ACT 1952 COMPLETE SPECIFICATION 6081 4

(ORIGINAL)

FOR OFFICE USE Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Tli- ll' *t co--t. ills te amendwcnts made under1 ctin 49 and is correct for haieg 11 Priority: Related Art: c r TO BE COMPLETED BY APPLICANT S- ''Name of Applicant: Address of Applicant: Actual Inventor: Address for Service: DR. HORST KIEF Londoner Ring 105, D-6700 Ludwigshafen, Germany Dr. Horst Kief ARTHUR S. CAVE CO.

Patent Trade Mark Attorneys Goldfields House 1 Alfred Street SYDNEY H.S.W. 2000

AUSTRALIA

Complete Specification"for the invention entitled AN IMPROVED

METHOD.

The following statement is a full description of this invention including the best method of performing it known to me:- 1 ASC 49 3074F/NNG I- 7 i Wffi esSM^.

1' r~ 0* 0 004 0 9 0* 5 *0 S 0 5 The invention relates to a process of the kind defined in claim 1 and to the use of the substance produced by this process, referred to for brevity as a "suspension", in particular to influence the immune system in the human and anima'l organism.

In the present state of medical science and technology there are various ways of influencing the immune system of the body.

An example is by passive or active vaccination, i.e. by the stimulation of antibodies or by the direct introduction of antibodies, whereby either suppressive or stimulatory effects can be produced on the immunomodulatory processes (definitions of medical terms in particular according to "Pschyrembet" clinical dictionary, Walter de Gruyter, Berlin-New York, 1972).

Particular importance has been achieved by the socalled desensitisation, in which the triggering antigen is first introduced to the organism'at a very high dilution and then in increasing doses, so as to neutralise the excessive antibody reactions.

Disadvantages of this process are the low success rate, the very prolonged treatment time, and the relatively limited area of indication of allergic illnesses.

A simple yet successful process is the injection of autologous blood. One such process is known in which dilutions of autologous blood are treated with suspensions of aluminium hydroxide, analogously to the binding of vaccines on aluminium hydroxide in long known processes, whereby the binding of the protein on the oxide certainly produces not only a certain deposit effect for the erythrocyte material and the plasma 1 2 proteins, but also a partial alienation of immunerelevant proteins.

This process has not achieved any great importance either, since it does not work reliably in the wider field.

Another method that should also be mentioned is an external method in which the patient's blood is very strongly oxidised by an ozone-oxygen mixture and then (i iCreturned to the organism. Individual observations r 10 indicate that here too immunomodulatory processes are t triggered off.

The use of suspensions for oxidation therapy is also described in particular in German patent specifi:cation 31 09 691, which refers to the autohemotherapy of Wolff and more particularly to the hyperbaric ozone therapy using venous blood of Kief, for which a device for extracorporeal bactericidal S" treatment is disclosed there. The oxidation process of Wehrlie is also described.

0*90 In the proposals of Wolff and Kief blood from the patient's own body, and this alone, is used as the starting substance and subjected to an oxidation treatment and then reinfused as such. This is thus .4 done in an exclusive manner, and there is only one oxidation step.

The invention is based on the observation that oxidised autologous blood can, in some cases, have a suppressive effect while in other similar cases it can have a stimulatory effect. Elucidation of this contradictory observation led to the discovery that oxidised or ozonised plasma often has an immunosuppressive effect while a similarly treated erythrocyte concentrate just as often has an immunostimulatory effect. But opposing processes can also be triggered off. Further unravelling of what was taking place led to the discovery that even 3 deproteinised serum can still trigger off immunomodulatory processes.

Even urine, which can be regarded in this connection, in a somewhat simplified manner, as an electrolyte solution that has been filtered off from the blood through the kidneys and selectively concentrated can, when administered by injection, trigger off immune processes.

o While the use of urine as a "desensitising agent" 10 was formerly often practised in naturopathic medicine, it has since been forgotten, as the disinfecting additions usually employed in the past, namely thymol and phenol, are themselves somewhat toxic and are unsuitable for use as additions. Urine cannot however be used unfiltered and unsterilised because of the risk of needle abscesses.

Based on these discoveries it is proposed, 'according to the invention, to fractionate the starting substance, which besides human or animal blood can also t l 20 be a tissue or a starting substance multiplied by oculturing, or even urine, in the process referred to above.

t The process according to the invention will now be 'i described, and developments of the invention will be mentioned.

The starting substance, for example the blood of the sick person, is in known manner aspirated into a sterile plasma bottle and heparinised to prevent coagulation. The amount of heparin added depends of course on the amount of blood aspirated. Residual air is then sucked out of the bottle and an ozone-oxygen' mixture is blown in under pressure, again in known manner, for example as described in the abovementioned patent specification. As mentioned above, tissue and/or urine can be used as the starting substance instead of blood. The latter likewise meets i:l all the requirements for a non-toxic, well-tolerated and above all effective immunomodulant.

The cause of the surprising effect of the invention may be found in the following ideas.

1. The cellular constituents of the human and animal blood, namely white and red blood corpuscles and thrombocytes, are embedded in the protein of the plasma, which in turn is dissolved in serum, the whole <ic thus forming an equilibrated and buffered system that 10 protects and maintains itself according to the S principles of regulation and counter-regulation. If S~the blood is now fractionated, the mutually regulating r" protective mechanisms of the individual systems break down and the selected fraction, for example washed erythrocytes, is directly exposed, without its protective coating, to a noxin, so that a controlled partial alienation of the proteins of cellular .4 constituents of the blood can exert a considerably stronger immunomodulatory effect.

20 2. The addition of ozone or other (strong) oxidants for immunomodulatory alienation of blood or 1 blood fractions may be regarded as a process that is very close to what occurs in nature, since the body i itself often falls tbck on oxidative processes as part of its natural defence mechanisms, for example the white blood corpuscles in defence against infection in the context of the "respiratory burst" by means of oxygen radicals, by which is to be understood the part of the body's own defence that takes place by means of oxidation.

3. Most stimulants, bacteria or viruses, produce an organotropic effect, i.e. in relation to our organ "blood" they only occupy particular constituents thereof, for example the Aids virus a particular lymphocyte sub-population, so that only after fractionation of the blood does the virucidal, C r C a Ote s a a a Iota a I (l

C

It C fungicidal and bactericidal effect of the oxidant Lead to an optimal and specific destruction of antigens and thus to specific immunostimulation.

4. The cells can defend themselves from the strong oxidising attack through particular enzyme systems in the cell membrane. Only after breaking up and/or destruction of the structure are very many important immunomodulatory substances accessible to oxidation. This break-up and destruction of protective structures can occur through mechanical influences (homogenisation), non-physiological temperatures (freezing), osmosis or proteolytic enzymes, e.g.

pepsin, papain or bromelain. The resulting "fracture sites" of particular protein fractions, with new potential points of attack for ozone or some other oxidant, emphasise the importance of repeated oxidation of the blood or of selected blood fractions. What has been said here in connection with "blood" also applies, within the scope of the invention and expressed in a 20 simplified manner, to the other starting substances mentioned.

In the oxidation it is of only subsidiary importance whether the oxidant is supplied to the medium concerned as a finished product from a Siemens discharge tube or whether for example it is generated in the material being treated itself by saturation with oxygen and possibly repeated irradiation with UV light.

If we consider the organotrophy of different antigens and of the associated antibodies in the different systems, the next step of a specific immunostimulation, with a possibly likewise specific selective immunosuppression in another organ system, becomes clear.

For this purpose it is proposed, after separate treatment of the fractions according to the invention, then to recombine either part or all of the 1

II

6 fractionated starting substance into a uniform suspension. In doing this it may be appropriate first to multiply the individual fractions by culturing them, for example a lymphocyte population as the carrier of particular antibodies. By the selective oxidation of a "blood fraction" multiplied in this way a concentrated antibody suspension is obtained that alienates in a "physiological manner" through the oxidant, and on renewed contact with the organism triggers a specific anti-autoantibody process that compensates the original pathological immunisation process. The renewed contact tF with the donor organism can be brought about parenterally, e.g. by injection, or orally, i.e. by swallowing, or even by inhalation or simple skin contact, e.g. by rubbing in.

Thus taking blood as an example, by subsequent r recombination of selected and separately oxidised (especially by ozonisation) fractions or hemolysates of the white and red systems of blood cells, of plasma, of defibrinated serum and/or of urine to a uniform dilution a medicament is obtained that has not only on .i the one hand the specific immunostimulatory properties of an active vaccine but also, at the same time and on the other hand, so far as is therapeutically appropriate, an immunosuppressive, organ-related effect, similar to that of a passive vaccine. By hemolysate is to be understood, in the sense of the invention, a suspension of plasma and intracellular fluid of the erythrocytes of variable composition.

A practical procedure is to centrifuge the erythrocytes out of autologous blood treated with an oxidant as described above, remove the plasma and treat it again with an oxidant, treat the erythrocytes again (after repeated washing) with an oxidant and partially suspend them in distilled water so as to cause them to burst through osmosis, and 'once again to treat them i i 7 with an oxidant. In particular cases a urine filtrate treated with an oxidant is added either to the serum or to the erythrocyte suspension. Ideally the desired hemolysate can be controlled by varying the oxygenozone concentration. It has been found that in the ideal case the concentration of the ozone-oxygen mixture is between 40 and 80 ng 03 per ml 02.

After recombination of the said fractions, possibly i c also with the urine filtrate, an extremely potent r 10 immunomodulant is obtained that has, depending on its t quantitative composition, both immunostimulatory and immunosuppressive properties. Proceeding in the manner 0€ indicated, for example with the blood serum of a particular group of patients, e.g. polyarthritics, a pooled serum can be obtained that can be used with O. success against rheumatism.

SBy the addition of carbonyl group carriers of aromatic or aliphatic structure when adding the oxidant, their sharply oxidising potential can be linked into the physiological paths, similarly to the 1 tquinones in the respiratory process. It is therefore proposed, as a further development of the invention, depending on the desired intensity of the oxidation Sprocess to add, at a particular point in the oxidant addition, e.g. ascorbic acid (vitamin this may be regarded as an ideal carbonyl group carrier of aromatic structure because of its conversion into dehydroascorbic acid during the chemical process. The conversion of vitamin C into dehydroascorbic acid can be stopped at any desired point in the process by freezing, particularly by shock freezing. Carbonyl group carriers of aliphatic structure can also be used.

In particular cases the immunostimulatory effect of the recombined suspension can be so strong that it should only be used diluted, e.g. 1:10, particularly in the initial phase of the treatment. It is particularly S t 1 1 1 1 advantageous to perform this dilution with ozonised water for injection: this is not only absolutely sterile but is also beneficial for a longer shelf life of the diluted suspension.

In a further development of the invention the addition of halogens, for example chlorine or iodine, to the suspensions described is of particular importance to the body's natural defences, e.g. by stimulation of a Haber-Weiss reaction and the resulting 10 prolongation of in vitro oxidative processes in vivo.

r In the known Haber-Weiss reaction a substance. e.g.

iodine, is oxidised to an unstable compound that supplies oxygen as a radical: the substance is again oxidised, the process begins from the beginning. This chain reaction continues until the oxygen present is used up. The added halogens can be in the form of acids and/or salts, also in complex form.

r t t 4 t L i 4 4

Claims

1. A process for the production of germicidaly treated and/or immunomodulatory active substances (suspensions) by an extracorporeal route, wherein the starting substances taken from a living sick person are subjected to an oxidation by means of ozone, oxygen and/or ionising radiation, characterised in that the starting substances are fractionated.

2. A process according to claim 1, characterised Sin that the starting substance is tissue.

3. A process according to claim 1, characterised ,in that the starting substance is multiplied through culturing.

4. A process according to claim 1, characterised r Ct f, in that the starting substance is urine.

5. A process according to one or more of the preceding claims, characterised in that the starting S' substance is comminuted to sub-cell size (cell lysis). Sr,'

6. A process according to claim 5, characterised in that the cell lysis is performed mechanically, 20 enzymatically and/or osmotically. .I

7. A process according to one or more of the preceding claims, characterised in that each of the fractions is separately filtered and again or only then subjected to the oxidation.

8. A process according to one or more of the preceding claims, characterised by the addition of carbonyl group carriers.

9. A process according to claim 8, characterised in that the addition takes place before the last oxidation.

A process according to claim 8 or claim 9, characterised in that the carbonyl group carrier is of aromatic structure.

11. A process according to claim 8 or claim 9, characterised in that the carbonyl group carrier is of aliphatic structure. 1 i

12. A process according to one or more of the preceding claims, characterised by repeated oxidation.

13. A process according to one or more of the preceding claims, characterised in the ozone content of the 02-03 mixture is controlled.

14. A process according to one or more of the preceding claims, characterised in that the ozone concentration of the 02-03 mixture is from 40 to 03 per ml 02.

15. A process according to one or more of the preceding claims, characterised in that ascorbic acid c is also added.

16. A process according to claim 15, characterised SI in that the ascorbic acid is added before the last oxidation step.

17. A process according to one or more of the preceding claims, characterised in that the starting substance or its fractions is or are diluted after the oxidation. t 20

18. A process according to claim 17, characterised rin that the dilution is performed with ozonised water.

19. A process according to one or more of the S preceding claims, characterised in that the treated it r Sfractions are recombined to a uniform dilution.

20. A process according to one or more of the preceding claims, characterised in that the finished suspension is shock-frozen.

21. A process according to one or more of the preceding claims, characterised in that the finished suspension is freeze-dried.

22. A process according to one or more of the preceding claims, characterised in that halogens, preferably chlorine or iodine, are added to the suspension.

23. A method for influencing the immune system in the human or animal organism which comprises administering to said body an effective amount of the suspensions produced according to any one of the preceding claims as pooled sera (for other persons).

24.. A method for influencing the immune system in the human or animal organism which comprises administering to said body suspensions produced according to any one of the claims l to 22 as a portable dilution. A method.for influencing the immune system in the human :e or animal organism which comprises administering to said body suspensions produced according to any one of claims 1 to 22 as S an inhalable solution. o oe 26. A mehtod for influencing the immune system in human or animal organisms which comprises administering to said body e 0 suspensions produced according to any one of claims 1 to 22.for rubbing in. 0 0. DATED this 10th day of December, 1990. c t| S DR HORST KIEF By Its Patent Attorneys ARTHUR S. CAVE CO. S 304 G -i; NI 11 3074F/NNG i