ZA200105929B - α, β, integrin inhibitors. - Google Patents

Description

) [4 PCT/EP99/09842

Inhibitors of the integrin o,f :

The invention describes novel peptides which, as ligands of the integrin a.fes, are biologically active.

These peptides all have a common structural motif, namely —- Asp Leu Xaa Xaa Leu -, Or in a preferred form - Arg Thr Asp Leu Xaa Xaa Leu Arg. The peptides according to the invention can be employed as effective inhibitors of the a,Be integrin receptor and thus for the treatment of various diseases and pathological findings.

Integrins belong to the Class I family of heterodimers - transmembrane receptors which play an important part in numerous cell-matrix and cell-cell adhesion processes (Tuckwell et al., 1996, Symp. Soc. EXP. Biol. 47). They can be roughly divided into three classes: the Pp; integrins, which are receptors for the extracellular matrix, the B, integrins, which are activatable on leucocytes and are “triggered” during inflammatory processes, and the oy .integrins, which . affect the cell response during wound healing and other ~ pathological processes (Marshall and Hart, 1996, Semin.

Cancer Biol. 7, 191).

The integrins asP;, aimBa, si, whi, avfs and abs all bind to the Arg-Gly-Asp (RGD) peptide sequence, e.g. in the natural ligand fibronectin. Soluble RGD-containing peptides are able to inhibit the interaction of each of these integrins with fibronectin. ouBs is a relatively rare integrin (Busk et al., 1982 J. Biol. Chem. 267(9), 5790), which is formed in increased amounts during repair processes in epithelial tissue and preferably binds the natural matrix molecules fibronectin and tenascin (Wang et al., 1996, Am. J. Respir. Cell Mol.

Biol. 15(5), 664). The physiological and pathelogical functions of oa.fs are still not precisely known; however, it is suspected that this integrin plays an important part in physiological processes and disorders

AMENDED SHEET

\ Sz - » of (e.g. inflammations, wound healing, tumours) in which epithelial cells are involved. Thus Bs is expressed on keratinocytes in wounds (Haapasalmi et al., 1996, J.

Invest. Dermatol. 106(1l), 42), from which it is to be assumed that in addition to wound healing processes and inflammations other pathological skin events, such as, for example, psoriasis, can also be influenced by agonists or antagonists of the said integrin. ops furthermore plays a part in respiratory tract epithelium (Weinacker et al., 1995, Am. J. Respir. Cell

Mol. Biol. 12(5), 547), =e} that appropriate agonists/antagonists of this integrin could be successfully employed in respiratory tract disorders, such as bronchitis, asthma, pulmonary fibroses and respiratory tract tumours. Finally, it is known that ofs also plays a part in the intestinal epithelium, so that appropriate integrin agonists/antagonists could be used in the treatment of inflammations, tumours and wounds of the stomach/intestinal tract.

Up to now, no low-molecular-weight inhibitor has vet been found which binds selectively to the of: integrin.

It was thus the object to find, in addition to the natural high-molecular-weight ligands and antibodies known hitherto, which are difficult to handle therapeutically and diagnostically, potent, specific or selective low-molecular- weight ligands for o,f, preferably peptides, which can be used for the therapeutic areas mentioned but also as a diagnostic or 3C reagent.

It has been found that the peptide compounds of the formulae presented below and their salts as soluble molecules exert an action on cells which carry the receptor mentioned, or when they are bound to surfaces are artificial ligands for ofPe-mediated cell adhesion.

They especially act as of¢ integrin inhibitors, particularly inhibiting the interactions of the receptor with other ligands, such as, for example, the

; ) PCT/EP99/09842 binding of fibronectin. This action can be demonstrated by the method which is described by J.W. Smith et al. in J. Biol. Chem. 265, 12267-12271 (1990).

The dependence of the origin of angiogenesis on the interaction between vascular integrins and extracellular matrix proteins is described by P.C.

Brooks, R.A. Clark and D.A. Cheresh in Science 264, 569-71 (1994).

It was furthermore found that the new substances have very valuable pharmacological properties together with good tolerability and can be employed as medicaments.

This is described in greater detail further below.

The peptide compounds according to the invention can furthermore be used in vivo as diagnostics for the detection and localization of pathological conditions in the epithelial system if they are equipped with the appropriate markers (e.g. the Dbiotinyl radical) according to the prior art. The invention also encompasses conjugates with other active compounds, such as. cytotoxic active compounds, as well as conjugates with radiolabels for radiotherapy or PET diagnosis but also fusion proteins with marker proteins © 25 such as GFP or antibodies, or therapeutic proteins such as IL-2.

The invention thus relates to peptide compounds of the formula I

W'-X' Arg Thr Asp Leu X’X'Leu X KE, -W* T in which: ) x', x3, x', x° X°* each independently of one another are an amino acid residue, the amino acids independently of one another being selected from a group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu,

Gly, Phe, His, Ile, Leu, Lys. Met, Nle, homo-Phe, Phg,

Pro, Ser, Thr, Trp, Tyr or val, and the amino acids mentioned possibly also being derivatized,

AMENDED SHEET

: v PCT/EP99/09842 . Ca

Ww? is selected from a group OH, OR, NHR, NR;, NH,

Wl is H or an acyl radical

R 1s alkyl having 1-6 C atoms, n is a number from 0-5 and m is a number from 0-10.

In the cases in which m or n assumes a value of greater than 1, the radicals X! and X°® can each independently of one another be identical or different.

According to the invention, those amino acids or amino acid residues are also encompassed which, starting from the natural amino acids, are derivatized, or are homologues or isomers thereof. The amino acid residues are customarily linked to one another via their a-amino and a-carboxyl groups (peptide bonding).

Those peptide compounds in which x* is an amino acid residue which was selected from the group consisting of

Asp, Glu, Arg, Lys, His and Tyr, and finally those peptide compounds in which x* is an amino acid residue which was selected from the group consisting of Ser, Tyr, : Thr, Gly and Val. _

The preferred compounds (for meanings or abbreviations see above and below) thus include those of the general formula II

W!'-x!Arg Thr Asp Leu X’X'Leu Arg XE? | II

AMENDED SHEET

) : PCT/EP99/09842 v Ce

Particularly preferred peptide compounds according to the invention are those of the formula V

Ww!-X',Arg Thr Asp Leu Asp Ser Leu Arg XE—W?2 v : and in this context in particular those of the formula

VI wl-x!,Arg Thr Asp Leu Asp Ser Leu Arg Thr X%.1-W? VI. :

Finally, the following individual compounds are particularly preferred, those also being included which are modified at the N and C termini: (a) H-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-Thr-Tyr-Thr-Leu-

OH

(b) H-Arg-Thr-Asp-Leu-Asp-Ser—-Leu~Arg-OH (c) Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-Thr-OH . (d) Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-Thr-NH; (e) H-Arg-Thr-Asp-Leu-Asp-Ser—Leu~-Arg-Thr-OH (f) H-Arg-Thr-Asp-Leu-Asp-Ser-Leu~Arg-Thr-NH; ] (g) H-Arg-Thr-Asp-Leu-Tyr-Tyr-Leu-Arg-Thr-Tyr-OH (h) Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-NH;

The abbreviations mentioned above and below stand for the radicals of the following amino acids:

AMENDED SHEET

; ®

Ala A alanine

Asn N asparagine

Asp D aspartic acid

Arg R arginine

Cys Cc cysteine

Gln Q glutamine

Glu E glutamic acid

Gly G glycine

His H histidine

Ile I isoleucine

Leu L leucine

Lys K lysine

Met M methionine

Nle norleucine

Orn ornithine

Phe F phenylalanine

Phg phenylglycine

Pro P proline

Ser S serine

Thr T threonine

Trp W tryptophan

Tyr Y tyrosine

Val \Y valine

If the abovementioned amino acids can occur in a number of enantiomeric forms, all these forms and also their mixtures are included above and below, e.g. as a constituent of the compounds of the formulae I-VI.

Furthermore, the amino acids for example, as a constituent of compounds of the formulae I-VI, can be provided with appropriate protective groups known per se.

The compounds of the formulae I-VI can have one or more chiral centres and therefore occur in various stereoisomeric forms. The formulae indicated include all these forms, in particular the D and L forms, especially in enantiomeric and racemic mixtures.

Finally, the formulae I and II mentioned above and below according to the invention also include the

. ® - 7 = corresponding salts, in particular the corresponding physiologically acceptable salts.

So-called prodrug derivatives are also included in the compounds according to the invention, i.e. compounds of the formula I modified with, for example, alkyl or acyl groups, sugars or oligopeptides, which are rapidly cleaved in the body to give the active compounds according to the invention. Furthermore, derivatives are also included in the compounds according to the invention which consist of the actual peptides according to the invention and known marker compounds which make it possible to detect the peptides easily.

Examples of such derivatives are biotinylated or fluorescence-labelled peptides.

In general, the peptides according to the invention are linear, but they can also be cyclized. The invention comprises not only the peptides of the formulae I to VI mentioned but also mixtures and preparations which in addition to these compounds according to the invention also contain other pharmacological active compounds or adjuvants which can influence the primary pharmacological action of the peptides according to the invention in a desired manner.

The compounds according to the invention and also the starting substances for their preparation are otherwise prepared by methods which are known per se and frequently employed, such as are described in the literature (e.g. in the standard works such as Houben-

Weyl, Methoden der organischen Chemie [Methods of

Organic Chemistry], Georg-Thieme-Verlag, Stuttgart), namely under reaction conditions which are known and suitable for the reactions mentioned. Use can also be made here of variants which are known per se.

Preferably, the peptides according to the invention can be prepared by means of solid-phase synthesis and subsequent removal and purification, as has been 3

! ® described, for example, by Jonczyk and Meienhofer (Peptides, Proc. 8% Am. Pept. Symp., Eds. V. Hruby and

D.H. Rich, Pierce Comp. III, p. 73-77, 1983, or Angew.

Chem. 104, 1992, 375), or according to Merrifield (J.

Am. Chem. Soc. 94, 1972, 3102). Otherwise, they can be prepared by customary methods of amino acid and peptide synthesis, such as are known, for example, from

Novabiochem - 1999 Catalog & Peptide Synthesis Handbook of Calbiochem-Novabiochem GmbH, D-65796 Bad Soden, from numerous standard works and published patent applications. Biotinylated or fluorescence-labelled peptides/proteins can likewise be prepared by standard methods (e.g. E.A. Bayer and M. Wilchek in Methods of

Biochemical Analysis, Vol. 26, The Use of the Avidin-

Biotin Complex as a Tool in Molecular Biology; and

Handbook of Fluorescent Probes and Research Chemicals, 6" Edition, 1996, by R.P. Haugland, Molecular Probes,

Inc.; or alternatively WO 97/14716).

Of course, the peptides of the formulae I-VI can also be liberated by solvolysis, in particular hydrolysis, or by hydrogenclysis of their functional derivatives.

Preferred starting substances for the solvolysis or hydrogenolysis are those which, instead of one or more free amino and/or hydroxyl groups, contain corresponding protected amino and/or hydroxyl groups, preferably those which, instead of an H atom which is connected to an N atom, carry an amino protective group or which, instead of the H atom of a hydroxyl group, carry a hydroxyl protective group.

The same applies to carboxylic acids which can be protected by substitution of their -CO-OH hydroxyl function by means of a protective group, e.g. as an ester.

The expression “amino protective group” is generally known and relates to groups which are suitable for protecting (for blocking) an amino group from chemical reactions, but which are easily removable after the ]

y ® - 9 = desired chemical reaction has been carried out at other positions in the molecule. The expression “hydroxyl protective group” 1s likewise generally known and relates to groups which are suitable for protecting a hydroxyl group from chemical reactions, but which are easily removable after the desired chemical reaction has been carried out at other positions in the molecule. The liberation of the compounds from their functional derivatives is carried out - depending on the protective group used - e.g. using strong acids, expediently using TFA or perchloric acid, but also using other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids such as trichloroacetic acid or sulfonic acids such as benzene- or p-toluenesulfonic acid. Hydrogenolytically removable protective groups (e.g. CBZ or benzyl) can be removed by treating with hydrogen in the presence of a catalyst (e.g. of a noble metal catalyst such as palladium, expediently on a support such as carbon).

The procedures are generally known and are not to be described in greater detail here.

As already mentioned, the peptides according to the invention include their physiologically acceptable salts, which can likewise be prepared by standard methods. Thus, a base of the formula I can be converted into the associated acid addition salt using an acid, for example by reaction of equivalent amounts of the pase and of the acid in an inert solvent such as ethanol and subsequent evaporation. For this reaction, suitable acids are in particular those which yield physiologically acceptable salts. Thus inorganic acids can be used, e.g. sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid, furthermore organic acids, in particular aliphatic, alicyclic, or araliphatic, aromatic or heterocyclic mono- or polybasic carboxylic, sulfonic or sulfuric acids, e.g.

. ® formic acid, acetic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acids, nicotinic acid, isonicotinic acid, methane- or ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, naphthalenemono- and disulfonic acids, laurylsulfuric acid. Salts with physiologically unacceptable acids, e.g picrates, can be used for the isolation and/or purification of the compounds according to the invention. On the other hand, an acid of the formula I can be converted into one of its physiologically acceptable metal or ammonium salts by reaction with a base. Possible salts in this case are in particular the sodium, potassium, magnesium, calcium and ammonium salts, furthermore substituted ammonium salts, e.g. the dimethyl-, diethyl- or diisopropylammonium salts, monoethanol-, diethanol- or diisopropylammonium salts, cyclohexyl- or dicyclohexylammonium salts, dibenzylethylenediammonium salts, furthermore, for example, salts with arginine or lysine.

The peptide compounds according to the invention can be employed, as already mentioned, as pharmaceutical active compounds in human and veterinary medicine, in particular for the prophylaxis and/or therapy of disorders in which epithelial «cells are involved.

Particularly to be emphasized in this context are disorders or inflammations or wound healing processes of the skin, the respiratory organs and the stomach and intestinal area, thus, for example, apoplexy, angina pectoris, oncoses, osteolytic illnesses such as osteoporosis, pathologically angiogenic illnesses such as, for example, inflammations, pulmonary fibrosis, ophthalmological illnesses, diabetic retinopathy, macular degeneration, myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, rubeoctic

. ® glaucoma, ulcerative colitis, Crohn's disease, atherosclerosis, psoriasis, restenosis after angioplasty, in acute kidney failure or nephritis.

The invention accordingly relates to peptide compounds of the formulae defined above and below and in the claims including their physiologically acceptable salts as medicaments, diagnostics or reagents.

The invention relates in particular to appropriate medicaments as inhibitors for the control of disorders which are based indirectly or directly on expression of the o.Bs integrin receptor, thus in particular on pathologically angiogenic disorders, thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, inflammations, infections and for influencing wound healing processes.

The invention also relates to appropriate pharmaceutical preparations which comprise at least one medicament of the formulae I to VI and, if appropriate, vehicles and/or excipients.

The invention furthermore relates to the use of the peptide compounds and/or their physiologically acceptable salts according to the claims and the description for the production of a medicament for controlling disorders which are based indirectly or directly on expression of the o,fs integrin receptor, thus in particular in pathologically angiogenic disorders, thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, inflammations, infections and for influencing wound healing processes. The medicaments according to the invention or pharmaceutical preparations comprising them can be used in human or veterinary medicine.

Possible excipients are organic or inorganic substances which are suitable for enteral (e.g. oral) or parenteral administration or topical application or for B

. ® administration in the form of an inhalation spray and do not react with the new compounds, for example water, vegetable oils, benzyl alcchols, alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum jelly. Tablets, pills, coated tablets, capsules, powders, granules, syrups, juices or drops, in particular, are used for oral administration, suppositories are used for rectal administration, solutions, preferably oily or aqueous solutions, furthermore suspensions, emulsions or implants, are used for parenteral administration, and ointments, creams or powders are used for topical application. The new compounds can also be lyophilized and the lyophilizates obtained used, for example, for the production of injection preparations. The preparations indicated can be sterilized and/or can contain vehicles such as lubricants, preservatives, stabilizers and/or wetting agents, emulsifiers, salts for affecting the osmotic pressure, buffer substances, colourants, flavourings and/or one or more further active compounds, e.g. one or more vitamins.

For administration as an inhalation spray, sprays can be used which contain the active compound either dissolved or suspended in a propellant or propellant mixture (e.g. CO; or chlorofluorohydrocarbons).

Expediently, the active compound is used here in micronized form, it being possible for one or more additional physiologically tolerable solvents to be present, e.g. ethanol. Inhalation solutions can be administered with the aid of customary inhalers.

The substances according to the invention can as a rule be administered in analogy to other known, commercially available peptides (e.g. described in US-A-4 472 305), preferably in doses between approximately 0.05 and 500 mg, in particular between 0.5 and 100 mg, per dose unit. The daily dose is preferably between approximately 0.01 and 20 mg/kg of body weight. The

J ® specific dose for each patient depends, however, on all sorts of factors, for example on the efficacy of the specific compound employed, on the age, body weight general state of health and sex, on the diet, on the time and route of administration, on the excretion rate, pharmaceutical combination and severity of the particular disorder to which the therapy applies.

Parenteral administration is preferred.

The invention finally also comprises recombinant DNA sequences which contain sections which code for peptide regions which contain the peptide structural motifs of the formulae I to VI according to the invention.

Such DNA can be transferred to cells by particles, as is described in Ch. Andree et al. Proc. Natl. Acad.

Sci. 91, 12188-12192 (1994), or the transfer to cells can be increased by other vehicles, such as liposomes (A.I. Aronsohn and J.A. Hughes J. Drug Targeting, 3, 163-169 (1997)).

The transfer of such a DNA could accordingly be used in yeasts, by means of bacculoviruses or in mammalian cells, for the production of the peptide substances of this invention.

If an animal or human body is infected with such a recombinant DNA, the peptides according to the invention finally themselves formed by the infected cells can bind directly to the ayfs integrin receptor, for example of tumour cells, and block it.

Appropriate recombinant DNA, which can be prepared by known and customary techniques, can, for example, however also be present in the form of virus DNA which contains sections which code for the virus coat protein. By infection of a host organism with recombinant, preferably non-pathogenic viruses of this

. ® type, host cells which express the integrin o,f can preferably be attacked (targeting).

Suitable viruses are, for example, adenovirus species which have been used a number of times already as vectors for foreign genes in mammalian cells. A number of properties make them good candidates for gene therapy, as can be inferred from S.J. Watkins et al.

Gene Therapy 4, 1004-1012 (1997) (see also J.

Engelhardt et al. Hum. Gene Ther. 4, 759-769 (1993)).

As can be found in A. Fasbender et al. J. Clin. Invest. 102, 184-193 (1998), the limited efficiency of the gene transfer is a common problem in gene therapy by viral and non-viral vectors. Using the above-described additional ligand sequence for a,fs integrin in the coat protein of the adenoviruses, an improvement in the transfer, for example, of cystic fibrosis transmembrane conductance regulator (CFTR) cDNA can be achieved.

Similarly to the work of T. Tanaka et al. Cancer

Research 58, 3362-3369 (1998), instead of the DNA for angiostatin the DNA for the sequences of this invention can also be used for cell transfections by means of retroviral or adenoviral vectors.

The peptides according to the invention can also be employed for use in gene therapy in man within a liposome complex of lipid/peptide/DNA prepared for transfection of cell cultures together with a liposome complex consisting of lipid/DNA (without peptide). The preparation of a liposome complex of lipid/DNA/peptide is described, for example, in Hart S.L., et al 1998:

Lipid-Mediated Enhancement of Transfection by a Non- viral Integrin-Targeting Vector, Human Gene Therapy 9, 575-585.

A liposome complex of lipid/peptide/DNA can be prepared, for example, from the following stock solutions: 1 upg/ul of lipofectin (equimolar mixture of i

Claims (25)

- w PCT/EP99/09842 CLAIMS

1. Peptide compounds of formula I W!-x! Arg -Thr Asp Leu X’X'Leu X’X5,-W? I in which: x!, x3, x*, X%X°, X5 each independently of one another are an amino acid residue, the amino acids independently of one another being selected from a group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Gly, Phe, His, Ile, Leu, Lys, Met, Nle, homo-Phe, Phg, Pro, Ser, Thr, Trp, Tyr or Val, and the amino acids mentioned possibly also being derivatized, w! is H or Ac, Ww? is OH, OR, NHR, NR,, NH,, R is alkyl having 1-6 C atoms, n is a number from 0-5 and m is a number from 0-10.

2. Peptide compounds according to Claim 1, in which x? is an amino acid residue selected from the group consisting of Asp, Glu, Arg, Lys, His or Tyr.

3. Peptide compounds according to Claim 1, in which X* is an amino acid residue selected from the group consisting of Ser, Tyr, Thr, Gly or Val.

4. Peptide compound according to Claim 1 as in the formula V W!-X' Arg Thr Asp Leu Asp Ser Leu Arg X°p-W V AMENDED SHEET -

; 4 PCT/EP99/09842 having the meanings indicated in Claim 1.

5. Peptide compound according to Claim 4 as in formula

Iv. W!-X! Arg Thr Asp Leu Asp Ser Leu Arg Thr X°,-w* VI

6. Peptide compounds of the formula I or II according to Claims 1 to 5 and their physiologically acceptable salts as medicaments.

7. Medicament according to Claim 6 as an inhibitor for the control of disorders which are based on an expression and pathological function of of; integrin receptors.

8. Medicament according to Claim 7 for use in a method for the control of thromboses, cardiac infarct, coronary heart disorder, arteriosclerosis, tumours, osteoporosis, fibrosis, inflammations, infections, psoriasis and for influencing wound healing processes.

9. Pharmaceutical preparation comprising at least one medicament according to one of Claims 6 to 8 and, if appropriate, vehicles and/or excipients and, if appropriate, other active compounds.

10. Use of peptide compounds according to Claims 1 to 5 and/or their physiologically acceptable salts for the production of a medicament for controlling disorders which are based on an expression and pathological function of «8; integrin receptors.

11. Use according to Claim 10 for the production of a medicament for controlling thromboses, cardiac AMENDED SHEET i

. 4 PCT/EP99/09842 infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, fibrosis, inflammations, infections, psoriasis and for influencing wound healing processes.

12. Recombinant DNA comprising a sequence which codes for a peptide section which corresponds to a peptide compound of Claims 1 to 5.

13. Recombinant virus DNA according to Claim 12.

14. Virus, characterized in that it possesses a coat protein which has a sequence which corresponds to a peptide compound of Claims 1 to 5.

15. Use of a virus according to Claim 14 for the production of a medicament for controlling disorders which are based on an expression and pathological function of o,8; integrin receptors.

16. A substance or composition for use in a method for controlling disorders which are based on an expression and pathological function of a,8s, said substance or composition comprising a virus according to Claim 14, and said method comprising administering said substance or composition.

17. A compound according to Claim 1, or Claim 6, substantially as herein described and illustrated.

18. A medicament according to Claim 7, substantially as herein described and illustrated.

19. A medicament for use in a method of treatment according to Claim 8, substantially as herein described and illustrated. AMENDED SHEET _

- | 4 PCT/EP99/09842

20. A preparation according to Claim 9, substantially as herein described and illustrated.

21. Use according to Claim 10, substantially as herein described and illustrated.

22. DNA according to Claim 12, or Claim 13, substantially as herein described and illustrated.

23. A virus according to Claim 14, substantially as herein described and illustrated.

24. Use according to Claim 15, substantially as herein described and illustrated.

25. A new compound, a new medicament, a new preparation, a new use of a compound according to any one of Claims 1 to 5 and/or of a physiologically acceptable salt thereof, new DNA, a new virus, a new use of a virus according to Claim 14, or a substance or composition for a new use in a method of treatment, substantially as herein described. AMENDED SHEET -