DE19757932C2 - Oligopeptides - Google Patents

Description

The totality of living organisms, from the lowest to the most organized, is constantly threatened by viral infections. There are currently 7,000 viruses known. 500 of them are human pathogenic and cause direct infectious diseases, some of which are incurable is.

Viral infections, including those that appear to be problem-free from the defense systems overcome, sometimes leave profound changes in the genome and Metabolism of the host.

In particular, viruses can induce tumor formation and cell destruction by auto- Cause immune reactions.

Furthermore, viruses and virus-assimilated particles are slow to develop evolving diseases involved, such as multiple sclerosis, Parkinson's, Creutzfeld Jacob as well as cardiovascular syndromes (cardiomyopathy), which after long latency periods Breakout come.

Diseases such as Alzheimer's as well as genetic diseases and trisomy (Mongolism) seem to involve viral activity.

Unexpectedly, viruses have recently been identified as the cause of other, completely different diseases:

• - Adenovirus 36 causes a particularly severe, untreatable Form of obesity
• - The equine Bornavirus can cause behavioral disorders, depression in humans and trigger manic-depressive insanity
• - For certain forms of juvenile diabetes, a type C retrovirus shows responsible.

Contrary to previous beliefs, the viral origin of such diseases also includes theirs transferable character. The transmission can take place horizontally and / or vertically. Clinical names related to such and other serious Diseases, expressions such as "familial clustering" and "genetic pre-disposition" are from the point of view of an increased viral load, which is in the life cycle of the data subjects has spread to recheck.

With the current state of knowledge in virology it can be assumed that the Most of the serious and with the current state of the art (Antibiotics) short-term or non-curable diseases or viral or para-viral activity for longer periods as a trigger. The morphological, genetic and / or Metabolic dysfunctions are then a late consequence and end result of a virus intrusion consider. The mistake was made repeatedly, the spectacular end results to consider such Dys function series as their cause (see prion protein in BSE, that, in reverse of a series of events that has not yet been understood, is autonomously replicative Pathogen properties can be assigned). This leads to unsuccessful therapy approaches and to neglect prevention. In fact, the virus is found in humans, animals and plants at the center of all kinds of diseases.

Viruses are spread horizontally by infection from individual to individual or vertically genetic takeover of endogenous viruses transmitted from the genomes of the ancestors.

Endogenous viruses transported in the genome over long generations can also Take over useful functions. For example, endogenous retroviruses can develop resistance be involved or take over the function of damaged genes in whole or in part.

Endogenous viruses can be activated, for example by other viruses, by Transposons or by all kinds of stress factors.

Numerous viruses can be transmitted through the air we breathe: flu and rhino viruses, adeno and Papilloma tumor viruses; at high concentrations even Lyssa viruses (rabies). The quick one or slow spread of such viruses can lead to worldwide epidemics. Died in 1919 20 million people with the A H1N1 influenza virus. Approved vaccines have so far existed only against 25 human pathogenic viruses.

Alimentary and iatrogenic accidents can cause viral infections of an unmistakable extent cause. In an enormous vaccine accident in England, the cause of the scrapie Disease of the sheep spread worldwide and then through negligent species passage first transferred to cattle (BSE) and then across the entire mammal area including the kidnapped.

Much of humanity was in the sixties at the worldwide Vaccination campaign against the poliovirus simultaneously accidental in the Simian Cell cultures introduced highly active and non-eliminable SV 40 tumor virus inoculated, which then in multiple variations in the affected individual and in the Successor generations became effective. With the explosive spread of The SV 40 is involved in tumor diseases.

The concentration of humans and animals, misery and absence of hygiene in the third world, the modern means of transport and the unlimited World trade with its sacred priority over all other considerations of caution and precaution  Reason ensure that the viral pathogen becomes more and faster Transmission vectors are available.

But not only viruses are transported, transmitted, mixed and recombined faster.

Also the mixture of terrain through alternating passages of the pathogen over different Breeds can help increase virulence. As is well known, the Eppstein-Barr Virus (EBV) in adolescents of the white race, Pfeiffer's glandular fever (infectious Mononucleosis), Burkitt sarcoma in African children and Asian children Adults have a rhinopharyngeal carcinoma. In mixed-race societies, this occurs for some time also Burkitt sarcoma in whites: The permissivity for the viral Symptom expression has expanded.

On the other hand, they cause environmental degradation, food toxicity and their Depletion of plant secondary metabolites, which are ultimately found in plants and animals Defense against parasites, an increasing weakening of the immune systems of the affected Host organisms, especially humans.

More and more and more virulent viruses are increasingly defensible hosts across from.

The situation evolves towards a collective immune collapse of all higher ones Creature.

Finding substances with selective antiviral activity without cytotoxicity and with high resistance to premature body degradation is a question of survival for humans and animal on the increasingly overpopulated globe.

Such substances could be predominantly chemically-synthetically oriented Research has so far not been delivered despite enormous search work.

The discrepancy between the enormous advances in knowledge of detection, the Morphology and the genetics of viruses on the one hand and the clinical helplessness the virus-infected individual, on the other hand, has never been so large.

The long-standing lack of success in efficient virus control without damage to the unaffected host cells is best characterized by the current situation in clinical AIDS treatment. The currently available tritherapy for HIV 1 and 2, consisting of two different nucleoside analogues such as AZT and 3 TC as inhibitors of reverse virus transcriptase, combined with an inhibitor of the viral protease (indinavir, ritonavir or saquinavir), allows a temporary reduction the viral load in more or less advanced stages of the disease

• - but only affects a part of the patients who have a responsive Virus serotype dominates
• - sooner or later leads to the formation of resistance through the appearance aggressive, untreatable virus strains that occur during medication selective selection run in the host the strains eliminated by the therapy replace and finally control the terrain of the infected for themselves.

The transmission of such virus strains to third parties leads to particularly fulgurant disease courses in the newly infected

• - is also accompanied by serious clinical side effects, the End effects are not yet foreseeable today
• - To some of the sick under the current conditions measured comfortably and with a maximum remaining life until death therapy is an enormous expense (medication against the virus itself and against opportunistic infections) and highly qualified work performance (ongoing PCR dosing of viruses, Sequencing of the newly formed virus variants and their Proteins) required, a "luxury" that for the majority of the approx. 100 million people infected with AIDS should hardly be up for debate.

The problem with post-infection therapy against the others is also unsolved known 500 human pathogenic viruses to be directed.

The situation with epidemic neurodegenerative diseases is even more opaque like BSE, whose pathogens have so far not been lacking adequate detection methods could be found.

One step towards solving the problem: antiviral oligopeptides with a new structure and surprising properties

In the systematic search for antiviral natural substances, which have an effect at the plant / insect interface, particularly in the area of life and in the products of the social insect Apis mellifera, a previously unknown, simple substance class was found that is central to the antiviral defense strategy of this highly populated life system (50,000 individuals per 100 dm ³ ) is correlated.

In general, such substances are involved in epidemic protection throughout the bee state and ubiquitous there, such as B. in the hemolymphs of insects and their metamorphic stages of development (larva, pupa, nymph) as well as in the embryonic Food juices for the 3 classes of workers, drones and queen.

The compounds discovered can be enriched and available in larger quantities in the Queen bee's lifelong feed juice (royal jelly) occur from where the discovery and initial isolation according to the invention also took place.

Condition for the appearance of these compounds in quantities as for the Characterization and assignment of functions from the raw substance (royal jelly) What was required is that the bees preferentially selectively base the bees certain types of plants are produced.

In fact, the Gelées Royales are derived mainly from plant pollen from selectively introduced into the working bees and from the allaitation bees to an emulsion of highly complex composition can be processed. This happens in the different Secretion glands from Apis mellifera, the most powerful glandular system in terms of dimensions represents all known living beings.  

Apis mellifera is a flying enzyme factory, where the plant constituents collected are immediately processed further. The highest mechanical, biochemical and reproductive performance as well as efficiency in the defense against parasites are combined in one organism. Apis mellifera is probably the best, evolutionarily oldest and longest proven system for solving the task according to the invention:

• - to eliminate viral pathogens without side effects,
• - or at least to suppress their symptoms so far that the infected host in the presence of the virus which has been rid of its virulence can realize a normal life and reproductive cycle
• - to ensure that the pathogens are not caused by resistance and reclaim the terrain by reviving virulent tribes.

Royal jelly is the key substance for the functioning of such a system.

To find the antiviral active substances according to the invention, the first step was optimal starting products wanted.

Selection of raw materials

60 quasi-monofloral Royals were made on the basis of 60 different ones bees.

Their effects were in the beehive itself, in which permanent inapparent viral infections without or with only weak symptoms, observed.

Then the antiviral activity was derived from monofloral sources Raw substances tested in vitro, in vivo and in the human field.

Among the monofloral money, 2 of them showed surprisingly high selectivity antiviral activity from: those that Crataegus oxyacantha (hawthorn) and Genista tinctoria (Dye broom) as a pollen and thus as a protein source. A row of other plant species followed at a certain distance: Brassica nappus, Taraxacum officinale, especially numerous Raphanus species, mustard plants and other cruciferous vegetables, almost only wild plants.

The royal jelly varieties thus obtained proved to be so unusual Composition that they are in the usual food standard Examination methods designated as "non-compliant" and not under this name Have been approved for sale. Such raw products are obviously in routine Analysis of food chemistry has not yet occurred.  

Test systems used

Virus / host systems were used both in vitro and in vivo as test systems with the help of which the selectively high antiviral activity of the royal jelly varieties obtained from hawthorn and gorse pollen were found:

• - Coxsackie B3 virus (Picorna virus) / Hela cells in the classic plaque essai: at a concentration of 200 ppm royal jelly Total virus titer reduction (none Infection sites on the test plaque more)
• - Influenza virus A2 (orthomyxovirus, pandemic flu,) / mouse: With peritoneal application of 200 mg jelly / kg body weight, one Survival rate of 100% for those fatally blind in mice ongoing infection.
• - Vesicular stomatitis virus (rhabdo virus) / mouse: The survival rate could be reduced by peritoneal, pre- and / or post-infection Application of the described royal jelly in concentrations of 1-500 mg / kg Body weight can be increased significantly, with Ginster jellies being an intermediate amount of 50 mg / kg had a clear maximum effect.
• - In specific human applications, symptom suppression and spontaneous remission were observed in the following viral infections:
  Influenza B (orthomyxovirus, non-pandemic flu)
  Herpes 2 virus (herpes virus)
  Hepatitis B virus (Hepadna virus)
  to some extent certain retroviruses
  HTLV-2 virus (hairy cell leukemia)

With these experiments, 6 are the main 24 currently known human pathogens Virus families covered.

Finally, the beehive itself was used as a virological test and experimental system:

• - The single bee is in most cases permanently inapparent with the cerebral infected with numerous viruses, especially with the ubiquitously widespread paralysis Virus APV (Picorna virus), without there being any under normal conditions Symptom expressions are coming. If the beehive is pollen-maternity leave, d. that is The starting product for obtaining the antiviral protein molecules is no longer available Or are caused by parasites (the mite Varroa jacobsoni) from the Removing the normally protected bee's hemolymph breaks through this virus activated the insect's cerebral gland barrier and infected via the protein-depleted juices the larvae. The entire beehive is from the Epidemic.
• - Re-feeding pollen and / or eliminating the hemolymph-sucking parasite leads to the immediate termination of the epidemic and a rapid regeneration of the  Bee colony. Certain types of pollen are more efficient than others. The effect of Pollen in the beehive correlates with the effects achieved in vitro and in vivo corresponding royal jellies obtained therefrom.

Obtaining the raw materials

To obtain the particularly antiviral active royal jelly described a total of 50 beehives in the usual way on the in the professional state of the art Beekeepers repurposed well-known queen rearing (separation grid for queen, 20 to 60 larval-infected queen cells per stick). The sticks were in at the appropriate time the vegetation to be placed, taking into account the absence of simultaneous flowering, bee-attractive competitive cultures. 10 to 120 hours, preferably 72 hours, after the queen larvae have been bred, those in the Aspirated jelly contained cells.

In order to optimally preserve the antiviral properties, it is suitable for all jellies Royals proved to be advantageous in any case, the suction from the queen cells and the Storage (5 ° C) under protective gas, e.g. B. Argon (Patent No. DE 42 32 732, EP 0 663 831, PCT 93 025 62, USA 5 580 297) and to the usual freezing or Refrain from lyophilizing.

The authenticity of the vegetable origin was checked and confirmed by pollen-morphological analysis of the royal jelly varieties thus obtained. Since gorse flowers only have a limited attraction for the flying bees, they always collect only a limited but regular proportion of them. This is reflected in the naturally low content of gorse pollen raw material in the rearing secretions (royal jelly). A two-phase process was used to obtain a royal jelly variety based almost exclusively on gorse dress:

• - First, with the help of normal beehives with pollen wipers Several kg of gorse pollen extracted, which mostly took place in remote wild regions
• - this material was obtained in a honey solution suspension of pollen-coated royal jelly- Bee colonies offered, which processed it without residue.

In this way, a royal jelly variety with one made almost entirely of gorse pollen resulting protein fraction obtained as it is naturally not attainable.

Simultaneously, however, there was an enrichment of broom alkaloids, which the therapeutic use of the royal jelly raw material thus obtained has certain limits sat.

Unused primary rock soils

Of particular importance for obtaining the active antiviral royal jelly varieties are the nature and nature of the soils, e.g. T. already through natural Location conditions for the selected plant species are defined.  

It was found that acidic, unused, pesticide-free granite Uraniciferous primary rock and constantly renewing soils are necessary. For gorse bushes and hawthorn hedges, such floors are the preferred ones Locations represent that the plants hardly grow on calcareous cultivated soils resulting gelées royales are of moderate activity.

Preliminary examinations indicated that certain, so far only with biologically negative Aspects contaminated trace elements such as cadmium and the elements of uranium and Thorium row when the described antiviral properties come about or for the plant biosynthesis of the substances causing them play an important role (Such metals have a highly specific effect, for example, in the recently discovered Metallo proteases).

A good supply of boron, which is inherently guaranteed in bedrock soils, plays a role also an important role (patent no .: DE 44 41 483, EP 95 93 82 85, PCT 95 044 94).

Furthermore, the climatic conditions are important: a high luminosity as well the absence of stressful conditions such as excessive drought or pesticide exposure promote the production of all types of secondary metabolites, including those antiviral oligopeptides according to the invention or their precursors are to be counted.

All of these conditions can be found in the ultimately selected test landscape united: a wide hedge landscape without large-scale agricultural use located in western France, between Cholet, Nantes and La Roche sur L'Yon. The area includes the Breton mainland plate and the bedrock of the Vendee. There are regular rainfall throughout the year. Still, the annual Sunshine duration high and comparable to that of the Mediterranean climate. The climate is mild and balanced: there is neither extreme heat nor extreme frost. The proximity to the Atlantic with the prevailing west winds ensure regular ventilation. The atmosphere is that of the Atlantic and not exposed to any local and cumulative environmental impact. The Wild flora is fully intact. All in all, it's probably a uniquely favored, apparently forgotten by the global environmental threat Biotope.

The plant species to be flown by bees are found in large-scale wild cultures Hedgerows and bush landscapes and with their roots reach through the soil the bedrock.

The granite rock base is rich in gneiss (amphibiols, biotite, plagioclase), Ferromagnesium, divalent cations of all kinds and sprinkling of pitchblende and Thorianite minerals.

The area contains several open pit mines for the extraction of uranium and thorium ores. The Natural radioactivity is high, but is largely discriminated against by plants royal jelly, pollen, honey, wax, propolis are not found in the bee products again.

The optimal environmental conditions for achieving the object according to the invention Plant and insect lead to one another in the royal jelly obtained Average conditions markedly changed, characteristic protein patterns.  

From the active raw product to the active molecule

From the raw products optimized according to the invention (= selectively obtained royal jelly), the task now was to identify and define the substances which are mainly responsible for the antiviral effects. For this purpose, the usual analysis methods for the investigation of complex natural product mixtures were used:

• - classic column chromatography
• - HPLC
• - one-dimensional electrophoresis
• - capillary electrophoresis

From this it was only possible to suggest the affiliation of the antiviral activity to one Protein fraction of low molecular weight can be recognized.

A detection peak in MALDI spectroscopy

The unambiguous assignment was finally made by means of low-molecular proteins specialized variant of MALDI mass spectroscopy, with the help of which ionized Molecules in the range from m / z 1000 to 10000 daltons with high selectivity and clearer Reproducibility can be quantified.

For this purpose approx. 1 mg was dissolved in 500 μl of water and with something for better solubility Acidic acid solution. From this clear solution, 2 ul with the appropriate Amount of matrix (DHAP) added. 0.5 µl of this was on the sample carrier crystallized. The sample prepared in this way could then be easily processed in the mass spectrometer be measured.

Since no fragmentation is observed with the MALDI-MS method, everyone corresponds Peak in the spectrum of a molecule.

In contrast to the higher molecular protein of royal jelly, which in large numbers (some 100) exist and are in species-related variations around a main storage protein group of 55000 D, there are only a dozen main Oligopeptides. Their relative amounts are approximately constant with the main peptides. Numerous analog, almost identical molecules, their concentration, exist like a satellite however in function of the vegetable raw material and the age of the royal jelly Emulsion fluctuates. This reasonably constant analysis scheme of the lower proteins was confirmed by 2-D gel electrophoresis.

There is one exception to this fixed rule in the peptide scheme: 5450 D is a good one characterized MALDI peak, the intensity of which varies from one royal jelly to another fluctuates extremely strongly. This selection peak is most pronounced in Spectra before that from native royal jelly with a high proportion of gorse, hawthorn and certain wild plant cruciferous pollen proteins (e.g. Raphanus species, mustard Plants) emerge. These are the same raw substances that the Develop the most intense antiviral activity. So the substance to be assigned to the peak is correlated directly or indirectly with the antiviral effect sought.  

An unusual oligopeptide of unusual purity

The selected raw jelly products could be displayed in large quantities, whereby part of it could be used for direct therapy attempts.

In contrast, the isolation of the target molecules prepared as in the previous analytical Search procedure great difficulty. This is due to the highly complex composition of the raw product: some 1000 proven or detectable compounds; all in all probably a few million z. T. ephemeral, mutually convertible trace components.

Indeed, royal jelly is like the composition of a living cell. This matches with its middle position as a post-cellular product of the plants and pre-cellular mixture for Formation of the insect embryos. The queen bee produces her 2 million offspring Made exclusively from royal jelly, with no condensation whatsoever Metabolic excretion is coming.

Classic methods such as side exclusion chromatography (Sephadex G 50), Ion exchange chromatography and one-dimensional electrophoresis led to Mixtures of substances with changing composition.

What was achieved in the analysis with MALDI differs as a separation method in the preparative scale.

A recently developed 2D electrophoresis technique has proven to be the method of choice. The first dimension is isoelectric focusing (IEF) with immobilized pH Gradients, followed by gel electrophoresis perpendicular to it.

In this way, the peptide was isolated in the highest purity, but this had to be bought with the disadvantages inherent in the method:

• - lowest amounts of substance
• - reduced biological activity

The procedure was as follows:

First, royal jelly is mixed with water (1: 2 ratio of royal jelly to water). Then the insoluble components are centrifuged off. The supernatant is made with Pharmalyte pH 3-10 ("for JEF", Pharmacia Biotech) and DTT (dithioerythritol) added. Here the amounts of these additives are each 1%.

For isoelectric focusing, 30 µl of this solution are placed on IPG strips (Immobilized pH gradient) applied to the anionic side. The IPG strips (4 mm wide, 18 cm long) are made of polyacrylamide, in which immobilization agents (Fa. Pharmacia Biotech) a gradient of pH = 4-9 is established. Care should be taken that the IPG strips have previously swollen for about 16 hours. The following is used as the source solution Mixture prepared: 0.5% CHAPS (= 3 - [(3-cholamidopropyl) dimethylammonium] - propane sulfate), 0.2% Pharmalyte (pH 3-10), 20 mM DTT and 8 M urea.  

Deionized water is used to prepare the anode and cathode solution. Interfering CO ₂ in the gas space of the electrophoresis chamber is bound by paper strips soaked with 3 M NaOH solution.

After isoelectric focusing (IEF), several IPG strips are made for 15 minutes each equilibrated with two buffer solutions. The buffer solutions (50 mM TRIS / HCl; pH = 8.8) contain 6 M urea, 30% glycerin and 2% SDS. The first buffer solution will be DTT (Final concentration 90 mM) and the second iodoacetamide (260 mM) separately. For the anode and cathode solution becomes a buffer solution with 1.4% glycine, 0.3% TRIS and 0.1% SDS used.

A horizontal SDS-PAGE (sodium n-dodecyl Sulphate-polyacrylamide gel electrophoresis) based on a T-15% polyacrylamide Separating gel and a T-6% collecting gel. TRIS / HCl serves as a buffer. Doing so the pH of the bulk gel was set to 6.8 and that of the separating gel to 8.8.

Electrophoresis is stopped before the bromophenol blue front reaches the wick. The Protein staining is done with Coomassie. This way you can see in the IPG strips Spots with highly enriched and highly pure protein can be detected. With the help of Calibration proteins of different molar masses could be determined based on the running distance pronounced spots a low-molecular protein of molecular weight 5540 Dalton was found, localized and isolated by "blotting" or extraction.

The protein isolated in this way was of unusual purity, no by-products could be detected. The determined primary structure is just as unusual as the purity. It was determined by total sequencing: 52 amino acids are combined in a chain in the following way:
Lys (1) -Thr-Ser - Ile - Ser (5) -
-Val-Lys-Gly-Glu-Ser (10) -
-Asn-Val-Asp-Val-Val (15) -
-Ser-Gln-Ile-Asn-Ser (20) -
-Leu-Val-Ser-Ser-Ile (25) -
-Val-Ser-Gly-Ala-Asn (30)
-Val-Ser-Ala-Val-Leu (35)
-Leu-Ala-Gln-Thr-Leu (40)
-Val-Asn-Ile-Leu-Tyr (45)
-Thr-Asp-Ser-Asp-Pro (50)
Met Arg (52)

The composition shows a protein chain without Cys, His, Phe and Trp.

Because with increasing sequencing, the signal levels of those cleaved from the protein chain and derivatized amino acid decrease continuously due to the sequencing technique safe sequencing generally does not exceed 40 amino acids. Nevertheless, the complete sequencing of the oligopeptide according to the invention, the is composed of 52 amino acids. However, the last 3 amino acids could cannot be identified with absolute certainty, so that possible homologs also can be biologically active.

The search for homologies to the specified sequence in the databases yielded no results Agreement with known sequences: the substance consequently has a completely new kind Structure.

The structural analysis reveals that hydrophilic and hydrophobic amino acid residues are approximate are present in the same number, but are distributed unevenly. A highly hydrophobic The central area (9 Val, 5 Leu, 4 Ile, 3 Ala) is flanked by two hydrophilic areas. What is striking is the high number of Ser residues (10), which has a good water-soluble character of the native oligopeptide explained. There is only one aromatic amino acid (Tyr 45). The given sequence leads to a molecular weight of 5414 D, in agreement with the estimated by the MALDI measurements and by the 2D electrophoresis separation values found using calibration proteins.

Despite the absence of stabilizing disulfide bridges, the peptide shows an extreme Resistance to all tested proteases and other enzymes: trypsin, chymotrypsin, Pancreatic elastase, thermolysin, pepsin, collagenase, enzyme Glu C, and even pronase and Proteinase K. The molar ratio enzyme / substrate (normally 1/1000) can be increased by a thousand times (1/1) without the slightest proteolysis occurring: the Despite its good solubility, the peptide according to the invention is in a physiological environment biologically exceptionally stable, which is good when used as a drug Transport properties in serum and intestinal tract guaranteed.

In addition to its biological stability, the good solubility of the invention Oligopeptides in their native form are a good prerequisite for their bioavailability.

The oligopeptide according to the invention proved itself to be non-toxic to humans even in high doses: In Experimental animals or test persons have mg- Get amounts injected or orally, without the slightest Side effect could be observed.

In order to be able to dispose of larger amounts of the peptide, its total synthesis was repeated a modified Merrifield method. The synthetic product was practical insoluble in water and showed no biological activity in this form. To make it the native To approximate the shape excreted from the apis mellifera secretion glands, the synthesis product was taken up in trifluoroethanol and then carefully in transferred aqueous medium (5 mg peptide in 5 ml water), whereby no more precipitation took place. Freeze drying gave a lyophilizate which was found in the presence of  Sodium bicarbonate (PH ≈ 8) excellently dissolved and in its biological and physicochemical properties approximated the native product.

2D gel electrophoresis of the low molecular weight proteins in the examined royal gels showed that the main peptide described is variable in small amounts in most cases is flanked by quasi-identical satellites. There are none in the molecular masses Find differences, but the isoelectric points differ by up to a few tenths of pH units. It appears that in the antiviral effect this Associate a function with a function in such a way that a kind of "oligo Therapy "makes it difficult for the viruses to evade by forming resistant strains.

It is proposed to use this nature-drawn scheme to use according to the invention by specifically modifying the pilot structure of the main peptide and is optimized for the biological effect against one or the other virus.

As a possible point of attack for the oligopeptide according to the invention and possibly its At the current level of knowledge, satellites on the viral cycle are a diminution closest to the effectiveness of the virus proteases.

Claims (4)

1. oligopeptide with antiviral activity, characterized in that it consists of at least 52 amino acid residues with the following sequence. 2. Oligopeptides according to claim 1, characterized in that amino acid residues can be replaced by others from position 48 of the sequence. 3. A process for the preparation of oligopeptides according to claims 1 and 2, characterized characterized in that gelées royales in the manner described among the specified Conditions are selectively obtained that contain these substances to a particular extent. 4. A method for activating a synthetic peptide according to claims 1 to 3, characterized in that it by gradually changing the solvent, followed by lyophylization and subsequent absorption in sodium hydrogen sulfate solution from an insoluble inactive in a soluble active, the biological form related conformation is converted.
Lys (1) -Thr-Ser - Ile - Ser (5) -
-Val-Lys-Gly-Glu-Ser (10) -
-Asn-Val-Asp-Val-Val (15) -
-Ser-Gln-Ile-Asn-Ser (20) -
-Leu-Val-Ser-Ser-Ile (25) -
-Val-Ser-Gly-Ala-Asn (30)
-Val-Ser-Ala-Val-Leu (35)
-Leu-Ala-Gln-Thr-Leu (40)
-Val-Asn-Ile-Leu-Tyr (45)
-Thr-Asp-Ser-Asp-Pro (50)
Met Arg (52)