EA004739B1 - PEPTIDE ANALOGS AND MIMETICS SUITABLE FOR IN VIVO USE IN THE TREATMENT OF DISEASES AS ASSOCIATED WITH ABNORMAL PROTEIN FOLDING INTO AMYLOID-LIKE DEPOSITS OR beta-SHEET RICH PATHOLOGICAL PRECURSOR THEREOF - Google Patents

Abstract

1. An inhibitory peptide capable of inhibiting beta pleated sheet formation in amyloid beta-peptide said inhibitory peptide wherein a peptide analog is a 5 residue Alzheimer inhibitor peptide iAbeta5 (SEQ ID NO:1 Leu-Pro-Phe-Phe-Asp) designed by chemical modification and achieved by a process selected from the group consisting of: alteration of the N-and C-terminal ends of said Alzheimer inhibitor peptide iAbeta5; replacing at least one residue of said Alzheimer inhibitor peptide iAbeta with alpha-aminoisobuiric acid (Aib); methylation of the alpha carbon of at least one residue of said Alzheimer inhibitor peptide iAbeta5; replacing at least one L-enantiomeric residue of said Alzheimer inhibitor peptide iAbeta5 with a D-enantiomeric residue, forming head to tail cyclization of said Alzheimer inhibitor peptide iAbeta5, replacing amide bonds in said Alzheimer inhibitor peptide iAbeta5 with an amide bond surrogate; and combinations thereof. 2. The inhibitory peptide of claim 1 wherein said alteration of the N-and C-terminal ends of said Alzheimer inhibitor peptide iAbeta5 is achieved by a process selected from acetylation, amidation, desamination, alcoholization and combinations thereof. 3. The inhibitory peptide of claim 2 wherein said inhibitory peptide is selected from the group consisting of: 4. The inhibitory peptide of claim 2 wherein said inhibitory peptide is ac-Leu Pro Phe Phe Asp-am. 5. The inhibitory peptide of claim 1 wherein said inhibitory peptide is selected from the group consisting of: 6. The inhibitory peptide of claim 3 wherein said inhibitory peptide is characterized by the following structure: 7. An inhibitory peptide capable of inhibiting conformational changes in prion PrP protein associated with amyloidosis which is an analog of 13 residue prion inhibitor peptide iPrP13 (SEQ ID NO: 2 Asp Ala Pro Ala Ala Pro Ala Gly Pro Ala Val Pro Val) designed by chemical modification SEQ ID NO :2, said chemical modification is achieved by a process selected from the group consisting of: alteration of the N-and C-terminal ends of said prion inhibitor peptide iPrP13; replacing at least one residue of said prion inhibitor peptide iPrP13 with alpha-aminoisobuiric acid (Aib); methylation of the alpha carbon of at least one residue of said prion inhibitor peptide iPrP13; replacing at least one L-enantiomeric residue of said prion inhibitor peptide iPrP13 with a D-enantiomeric residue, forming head to tail cyclization of said prion inhibitor peptide iPrP13, replacing amide bonds in said prion inhibitor peptide iPrP13 with an amide bond surrogate; and combinations thereof. 8. The inhibitory peptide of claim 7 wherein said alteration of the N-and C-terminal ends of said prion inhibitor peptide iPrP13 is achieved by a process selected from acetylation, amidation, desamination, alcoholization and combinations thereof. 9. The inhibitory peptide of claim 2 wherein said inhibitory peptide is selected from the group consisting of: 10. The inhibitory peptide of claim 2 wherein said inhibitory peptide is selected from the group consisting of: 11. The inhibitory peptide of claim 3 wherein said inhibitory peptide is characterized by the following structure: 12. A method for reducing the formation of amyloid or amyloid like deposits involving abnormal folding into beta sheet structure of amyloid beta peptide or for reducing the amount of said amyloid beta peptide which has already formed into a beta sheet structure comprising bringing into the presence of said amyloid beta peptide either prior to or after the abnormal folding thereof into a beta sheet structure, an effective amount of the peptide of claim 1. 13. A method for reducing the formation of amyloid or amyloid like deposits involving conformational changes in prion Pr protein or reducing the amount of said prion Pr protein which has already formed into amyloid or amyloid-like deposits comprising bringing into the presence of said prion Pr protein either prior to or after said conformational changes thereof into amyloid deposits an effective amount of the peptide of claim 7. 14. A method for reducing the formation of amyloid or amyloid like deposits by administration of a peptide selected from one of the group consisting of: 15. A pharmaceutical composition, comprising an inhibitory peptide capable of inhibiting beta pleated sheet formation in amyloid beta-peptide and a pharmaceutically acceptable carrier or excipient, said inhibitory peptide analog is a 5 residue Alzheimer inhibitor peptide iAbeta5 (SEQ ID NO: 1 Leu-Pro-Phe-Phe-Asp) designed by chemical modification and achieved by a process selected from the group consisting of: alteration of the N-and C-terminal ends of said Alzheimer inhibitor peptide iAbeta5; replacing at least one residue of said Alzheimer inhibitor peptide iAbeta with alpha-aminoisobuiric acid (Aib); methylation of the alpha carbon of at least one residue of said Alzheimer inhibitor peptide iAbeta5; replacing at least one L-enantiomeric residue of said Alzheimer inhibitor peptide iAbeta5 with a D-enantiomeric residue, forming head to tail cyclization of said Alzheimer inhibitor peptide iAbeta5, replacing amide bonds in said Alzheimer inhibitor peptide iAbeta5 with an amide bond surrogate; and combinations thereof. 16. The pharmaceutical composition of claim 15 wherein said inhibitory peptide is selected from the group consisting of: 17. The pharmaceutical composition of claim 15 wherein said inhibitory peptide is ac-Leu Pro Phe Phe Asp-am. 18. The pharmaceutical composition of claim 15 19. The pharmaceutical composition of claim 15 wherein said inhibitory peptide peptide is characterized by the following structure: 20. A pharmaceutical composition, comprising an inhibitory peptide inhibiting conformational changes in prion PrP protein associated with amyloidosis which is an analog of 13 residue prion inhibitor peptide iPrP13 (SEQ ID NO: 2 Asp Ala Pro Ala Ala Pro Ala Gly Pro Ala Val Pro Val) designed by chemical modification SEQ ID NO: 2, said chemical modification is achieved by a process selected from the group consisting of: alteration of the N-and C-terminal ends of said prion inhibitor peptide iPrP13; replacing at least one residue of said prion inhibitor peptide iPrP13 with alpha-aminoisobuiric acid (Aib); methylation of the alpha carbon of at least one residue of said prion inhibitor peptide iPrP13; replacing at least one L-enantiomeric residue of said prion inhibitor peptide iPrP13 with a D-enantiomeric residue, forming head to tail cyclization of said prion inhibitor peptide iPrP13, replacing amide bonds in said prion inhibitor peptide iPrP13 with an amide bond surrogate; and combinations thereof. 21. The pharmaceutical composition of claim 20 wherein said inhibitory peptide is selected from the group consisting of: 22. The pharmaceutical composition of claim 20 wherein said inhibitory peptide is selected from the group consisting of: 23. The pharmaceutical composition of claim 20 wherein said inhibitory peptide is characterized by the following structure:

Description

This application claims priority to provisional application US No. 60/163911, filed November 5, 1999.

BACKGROUND OF THE INVENTION

Field of Invention

The present invention relates to peptide analogues and peptidomimetics of peptides that violate the β-folding structure, suitable for the use of ίη νίνο in the treatment of mammals with diseases associated with a change in protein conformation, such as Alzheimer's disease and prion associated disease. More specifically, the present invention is directed to new peptide analogues and mimetics, pharmaceutical compositions containing one of such peptide analogues or mimetics, or a mixture thereof, as well as to methods for the prevention, treatment or diagnosis of disorders or diseases associated with abnormal folding of proteins in amyloid or amyloid-like deposits, or their precursors having a pathological β-folded structure.

Description of the Related Art

A large amount of data has been accumulated indicating that some of the diseases that are not similar in nature have the same molecular basis, i.e. a change in protein conformation (Tyoshak e! a1., Ttepbk Βίοс1ет. δα. 20: 456-459, 1995; δοΐο, 1. Μο1. Meb. 77: 412-418, 1999). Diseases associated with changes in protein conformation include Alzheimer's disease, prion-associated disorders, systemic amyloidosis, serpin deficiency disorders, Huntington's disease and amyotrophic lateral sclerosis (δο! Ο, 1999, above). A distinctive event in diseases associated with a change in protein conformation is a change in the secondary and tertiary structure of a normal protein without a change in the primary structure. A protein with a modified conformation can participate in the development of the disease due to direct toxic activity, loss of the biological function of a protein with normal folding or defective transportation (T1yusha8 e! A1., 1995, above). In cases where the protein is toxic, it usually self-associates, and it is deposited in the form of amyloid fibrils in different organs, inducing tissue damage (T1usha8 e! A1., 1995, above; Ke11u, Sigg. Ορίη. 81tis1. Βίο1. 6. : 11-17, 1996; δοΐο, 1999, above).

Alzheimer's disease (AD) is a debilitating neurodegenerative condition characterized by loss of short-term memory, disorientation and impaired ability to reason and think. AD is the most common form of dementia in the elderly. It is estimated that more than twenty-five million people all over the world are affected to varying degrees by AD (Terit, Atuyb 5: 121-142, 1998). A distinctive event in AD is the deposition of insoluble protein aggregates, known as amyloid, in the brain parenchyma and the walls of the blood vessels of the brain. The main component of the amyloid is a 4.3 kDa hydrophobic peptide called the amyloid beta peptide (Av), which is encoded on chromosome 21 as part of a much longer precursor protein (APP) (Ze ^ e, δс ^ еηсе 275: 630-631 , 1997). The genetic, biochemical and neuropathological data accumulated over the past 10 years allow us to confidently assume that amyloid plays an important role in the early pathogenesis of AD and, possibly, triggers the disease mechanism (δοΐο е! А1., 1. No. 63. 1191- 1198, 1994; δе1кое, 1997, mentioned above; Terit, 1998, mentioned above; δί8ο6ίη apb Rpsсе, ΡΑδΕΒ 1. 9: 366-370, 1995; δοΐο, Μο1. Furniture Shbau 5: 343-350, 1999).

Amyloid is a general term describing fibrillar aggregates that have a single structural motive, i.e. (β folding conformation ^ etre e! a1., Ce11 Μο1. b1 £ e δ ^. 53: 887, 1997; δ ^ ρе, App. Vet. Βίοсет. 61: 947-975, 1992). These aggregates have characteristic color properties, including the ability to emit birefringent green light after dyeing with Congox red, and the ability to bind fluorochrome, thioflavin δ (Ep, 1992, above; Οΐίκο е! А1., Μο1. No. igoyu1. 8: 49-64 , 1994). There are more than 12 human diseases of various etiologies, characterized by extracellular deposition of amyloid in different tissues, which leads to cell destruction, organ dysfunction and death. Among diseases including amyloidosis, one can distinguish Alzheimer's disease, prion-related disorders (also known as infectious spongiform encephalopathy) and systemic amyloidosis (Table 1). Amyloid fragments usually consist of proteolytic fragments of normal or mutant gene products. There are over 16 different proteins (Table 1) involved in amyloid deposits in various tissues (Ο1ίκο! A1, 1994, indicated above).

Amyloid formation is essentially a protein folding problem, as a result of which soluble peptides having mostly random folding become aggregated, taking (β-folded conformation (Ke11u, 1996, supra; δο! Ο, 1999, supra). The formation of amyloid proceeds with the participation of hydrophobic interactions between conformationally altered amyloidogenic intermediates, which are organized through peptide interactions into a β-folded structure. Hydrophobicity is an important factor for interactions of monomers leading to aggregation, while the β-folded conformation can determine the location of aggregates in amyloid fibrils. When trying to inhibit the formation of amyloid fibrils, these two properties were separated by constructing short synthetic peptides with sequence homology and the same degree of hydrophobicity as the peptide domain involved in conformational change, but having a very low tendency to form β-folded conformation (called peptides s destroying β-folds) (δοίο c1 a1, 1996, supra.; δοίο e1 a1., 1998, above). The goal is to construct a peptide that has the ability to specifically bind to amyloidogenic peptide, forming a complex that stabilizes the physiological conformation and destabilizes the abnormal conformation of the peptide (δοίο e1 a1., 1999, above).

Table 1. Diseases associated with amyloidosis and the protein component of amyloid fibrils

DISEASE

Alzheimer's disease

Primary Systemic Amyloidosis

Secondary systemic amyloidosis, familial Mediterranean fever

COMPONENT FIBRILL

Amyloid-β protein

Immunoglobulin light chain or fragments thereof

Fragments of serum amyloid-A

Spongiform encephalopathy

Senile systemic amyloidosis, familial amyloid polyneuropathy

Amyloidosis associated with hemodialysis

Icelandic Hereditary Cerebral Amyloid Angiopathy

Prion Protein Fragments

Transthyretin and its fragments

P2 microglobulin

Cystain C

Finnish family amyloidosis

Fragments of gelzolin

Type II diabetes

Fragments of an islet amyloid polypeptide

Familial amyloid polyneuropathy Fragments of apolipoprotein A-1 Thyroid medullary carcinoma Calcitonin Fragments Atrial Amyloidosis Atrial natriuretic factor Hereditary non-neuropathic systemic amyloidosis Lysozyme or wow fragments Hereditary renal amyloidosis Fibrinogen Fragments Islet amyloid Insulin Amyloidosis in aging Apolipoprotein Α-ΙΙ

To date, β-fold-destroying peptides have so far been designed to block conformational changes occurring in both Αβ and prion protein (Prg), which are involved in the pathogenesis of Alzheimer's disease and prion associated disease, respectively. In the prior art it was previously shown that peptides that destroy β-folds of 11 and 5 residues (namely, ίΑβ1 and ίΑβ5, respectively), homologous to the central hydrophobic region of Aβ, inhibit conformational changes in the peptides, leading to the formation of amyloid, and dissolve previously formed fibrils ίη νίίτο (δοΐο e1 a1., Vusyet. Vyuruk. Century. Sottii. 226: 672-680, 1996; δοίο e1 a1., No. 1 and Meb. 4: 822-826, 1998). In addition, a peptide of 5 residues is able to prevent the death of neurons induced by the formation of oligomeric β structures enriched with β folds in experiments with cell cultures (δοίο e1 a1., 1998, above). In addition, in the prior art, a rat model of amyloidosis induced by intracerebral injection of Aβ1-42 showed that joint injections of a peptide of 5 residues destroying β folds reduce the accumulation of Aβ in the brain and completely block the deposition of fibrillar amyloid-like plaques in rat brain (δοίο eί a1., 1998, above). Finally, a β-fold-destroying peptide introduced eight days after Aβ injection is capable of destroying ίη νίνο fibrils previously formed in the rat brain, which leads to a decrease in the size of amyloid deposits (δίβΐιπίκκοη. Egapscope. Δοίο, submitted manuscript). Interestingly, the removal of amyloid under the action of a peptide destroying β-folds returns to its previous state the cerebral histological damage associated with it, including contraction of neurons and activation of microglia.

The β-fold destroying peptides have also been designed to prevent and restore conformational changes caused by prions (Prg). Based on the same principles and when using the PrP sequence (114122) as a matrix, it was shown in the prior art that if a set of peptides destroying β-folds were synthesized, the peptide of 13 residues (1PrP13) had the highest activity (δοίο, 1999 indicated above). To test the inhibitory activity used several tests ίη νίνο and ίη νίίτο in cell cultures, and the results clearly demonstrated that it is possible not only to prevent the conversion of RGR ^with RGR ^CS, but also, more interestingly, return infectious conformational variant structure RGR ^ks in biochemical and structural state similar ^to WGR (δοίο eί a1., manuscript submitted for review).

Short peptides have been widely used in medicine as medicines (Vao eί a1., S. Wakata apb S.M. Αηaiίyya ^ ata ^ ay, ebk. Βοκίοη: Vykyaikeg, pp. 181-198, 1994). However, the development of peptide drugs is largely limited due to their insufficient oral bioavailability and the short duration of action resulting from the enzymatic degradation of ίη νίνο (Eaisige apb Tišteai, Α6ν. Aegis Century 23: 127159, 1992). The progress made in recent years in the production of peptide analogues (such as pseudopeptides and peptidomimetics) with a lower susceptibility to proteolysis increases the likelihood of obtaining useful drugs structurally related to the original peptides (Raieiege apb Tyipei, 1992, supra). Improving the resistance of the peptide to proteases not only increases the half-life of the compound in the bloodstream, but also increases its ability to transport or absorb at various levels, including absorption in the intestine and penetration through the blood-brain barrier, since transport and absorption in a high degree depend on the membrane contact time or barriers with bioactive samples (Raiyege apb Tyipei, 1992, above).

Summary of invention

The present invention provides an inhibitory peptide capable of inhibiting the formation of a β-folding structure in an amyloid β-peptide, wherein the inhibitory peptide is a β-folding disruption peptide analog designed by chemical modification of a β-folding disruption peptide capable of inhibiting the formation of a β-fold structure in amyloid β-peptide.

The peptide is chemically altered by: (1) modifications of the Ν- and C-terminus of the peptide; (2) side chain substitutions, which may include amino acid substitutions; (3) modifications on the α-carbon atom, including methylation, alkylation and dehydrogenation; (4) changing chirality by replacing the Ό-residue with an--residue; (5) head-to-tail cyclizations; and (6) introducing substitutions in the amide bond, i.e., replacing the atoms involved in the formation of the peptide (or amide) bond.

The present invention also includes an inhibitory peptide capable of inhibiting conformational changes in the Prg Prion protein associated with amyloidosis, wherein the inhibitory peptide is a β-fold destroying peptide analog designed by chemical modification of a β-folding destroying peptide capable of inhibiting conformational changes in a protein prions Prg associated with amyloidosis.

In addition, the present invention includes a peptidomimetic of the following structure:

has the following structure:

In another embodiment, the peptidomimetic has the following structure:

The present invention also includes a method for the prevention, treatment or diagnosis of disorders or diseases associated with abnormal folding of proteins in amyloid or amyloid-like deposits or their precursors having a pathological beta folding structure.

Brief Description of the Drawings

In FIG. 1 is a schematic representation of a peptide bond and potential target sites for peptide modifications.

In FIG. Figure 2 is a graph depicting the pharmacokinetics of the peptide inhibitor of Alzheimer's amyloidosis of 11 residues (eaten YAORRURURGO), which destroys (3 folds, in its natural L-configuration and in non-natural L-form.

In FIG. 3A and 3B show three-dimensional structures of peptides that destroy β-folds in Alzheimer's disease and the disease associated with prions, ίΑβ5 and 1PrP13, respectively.

In FIG. 4A and 4B are graphs showing the bioavailability and stability of ίΑβ5 and Αc- ^ Αβ5-Αt, respectively, over time.

In FIG. 5A is a graphical comparison of Aβ1-40 incubated with various other peptides.

On figv presents the dependence of the formation of amyloid on the concentration of Αο-ίΑβ5At.

In FIG. 5C shows the dependence of amyloid formation on концентрацииβ5 concentration.

In FIG. Figure 6 shows a model with 83% decay of deposits in the ventricle and 30% decay of amyloid plaque in the tonsil.

DETAILED DESCRIPTION OF THE INVENTION

In the present invention, the bioavailability and stability of the inhibitory peptide is improved by chemically modifying the parent peptide to give a more stable derivative when using ίη νίνο, which is preferably administered orally. An inhibitory peptide is capable of inhibiting the formation of a β-folded structure in an amyloid β peptide. Moreover, the inhibitory peptide is a peptide analog,

Ί suturing β-folds constructed by chemical modification of a β-folding destroying peptide capable of inhibiting the formation of a β-fold structure in an amyloid β-peptide.

The present invention also includes an inhibitory peptide capable of inhibiting conformational changes in the prion protein PrP associated with amyloidosis, wherein the inhibitory peptide is a β-fold destroying peptide analog designed by chemical modification of a β-folding destroying peptide and capable of inhibiting conformational changes in Prion Protein Protein Associated with Amyloidosis.

In FIG. Figure 1 shows a common peptide skeleton that shows possible targets for chemical modifications. Possible targets include the following: (1) modifications of the K- and C-terminal regions of the peptide (targets a and b); (2) side chain changes (target c), which usually include amino acid substitutions; (3) a modification at the α-carbon atom (target b), including methylation, alkylation and dehydrogenation; (4) changes in chirality by replacing the β-residue with a β residue; (5) head-to-tail cyclization; and (6) introduction of substitutions in the amide bond (target e), i.e. replacement of atoms involved in the formation of a peptide (or amide) bond. The latter derivatives are known as pseudopeptides or amide bond substitutes.

Natural peptides are usually destroyed as a result of the coordinated action of specialized endopeptidases and non-specific exopeptidases. Endopeptidases are often present in tissues and cell compartments and convert the peptide into two or more inactive fragments. Exopeptidases, as a rule, are present in the blood and peripheral organs and decompose intact peptides or their fragments into their amino acids, thereby contributing to the disappearance of peptides from the bloodstream. Exopeptidases recognize free amino or carboxyl groups in peptides. Therefore, modification of these groups often reduces or eliminates exopeptidase degradation. Cyclization of the head-to-tail peptide leads to the absence of free end groups and, therefore, also minimizes exopeptidase cleavage. Endopeptidases, on the other hand, recognize atoms involved in the formation of an amide bond. Thus, substitutions in the amide bond significantly reduce endopeptidase degradation. The same thing usually happens when modified at the α-carbon atom. Since most (if not all) exo- and endoproteases are stereospecific, substitutions of natural L-amino acids with стере-stereoisomers leads to an obvious increase in peptide stability. Finally, peptidomimetics are usually completely resistant to proteolytic degradation and can often be administered orally. Analogs that destroy β-folds obtained by chemical modifications of leader peptides

Modifications, starting with a peptide of 5 residues that inhibits Alzheimer's disease (ίΑβ5, last ΤΡΡΕΌ - also referred to as Ley Rgo Rye Pye Acre), and a peptide of 13 residues that inhibits prion associated disease (1PgP13, last EARAARASRAURU also denoted as Acre A1a Rgo A1a A1a Rgo A1a C1u Rgo A1a Ya1 Rgo Ya1), is carried out as described below. The peptides used in the present invention are synthesized using standard techniques disclosed by Vegdtapp e! a1. and incorporated by reference into this description (Vegtapp & 2guac, Riding Run EGGGGGGGGGGGGGGGQQGG (1932) 65: 1192-1201).

a) Modifications at the Ν- and C-terminus.

Ν-terminal acetylation or deamination provides protection against cleavage by a series of aminopeptidases, while the presence of a C-terminal carboxyl group substituted with amides or alcohols prevents cleavage by some carboxypeptidases, including carboxypeptidases A and B. Modified peptide sequences including these modifications are presented below where ac is acetylation, am is amidation, beck is deamination, and a1c is alcoholization.

Alzheimer's Disease Inhibitors:

as-bie rgo rne rne azr-ash des-bie rgo rie rie azr-at as-bie rgo rie rie azr-a1s without bie rogo rie rie azr-a1s

Prion associated disease inhibitors:

as-Azr A1a Rgo A1a A1a Rgo A1a C1u Rgo A1a Ua1 Rgo Ua1-at without Azr A1a Rgo A1a A1a Rgo A1a 31u Rgo A1a Ua1 Rgo Ua1-at as-Azr A1a Rgo A1a A1a Rgo A1a1a1a1a1a1a1a1a1a1a1a1a1a1a1a1a1a1a1a A1s aez-Azr A1a Rgo A1a A1a Rgo A1a C1u Rgo A1a Wa1 Rgo Wa1-a1s

b) Side chain changes.

The presence of unnatural amino acids usually increases the stability of the peptide. In addition to Gogo, at least one of these amino acids (α-aminoisobutyric acid or A1b) determines a significant strain on the designed peptides, reducing their conformational mobility. In particular, the inclusion of A1b in the β-folded model of peptides causes complete destruction of this structure. The activity of A1b, which blocks the formation of β folds, is comparable to or even exceeds the activity of the natural proline residue used in the peptide as a blocker for the formation of β folds. Therefore, the introduction

ΑΐΒ presumably increases both the stability of the peptide and its inhibitory activity.

Alzheimer's Disease Inhibitors:

Lye ΑΪΡ Rye Rye Azr

Prion associated disease inhibitors:

Azr A1a A1b A1a A1a A1b A1a A1a A1b A1a C1u A ± b A1a Ya1 A1b Ya1

c) Modifications to the α-carbon atom.

The most commonly used modification on the α-carbon atom, which is used to improve the stability of the peptide, is α-methylation. In addition, it was shown that the substitution of the hydrogen atom bonded to the α-carbon atom Pb, Va1, or Lb favors the adoption of a β-curved conformation and does not significantly contribute to the formation of β-folded structures. In accordance with the present invention, the methylation of these residues in inhibitory peptides is believed to increase stability and activity.

Alzheimer's Disease Inhibitors:

(Me) Te Rgo Rye Rye Azr

Lye Rgo (Me) Rye Rye Azr

Lye Rgo Rye (Me) Rye Azr (Me) Lye Rgo (Me) Rye Azr (Me)

Prion associated disease inhibitors:

Aar A1a Rgo A1a A1a Rgo A1a C1u Rgo A1a (Me) Wa1 Rgo Wa1

Azr A1a Rgo A1a A1a Rgo A1a 61u Rgo A1a Ua1 Rgo (Me) Ua1

Azr A1a Rgo A1a A1a Rgo A1a C1u Rgo A1a (Me) Ua1 Rgo (Me) Ua1

b) Changes in chirality.

Replacing the natural L-residue with Ό-enantiomers greatly increases the resistance to proteolytic degradation. An increase in stability as a result of the introduction of the Ό-residue has already been shown for a peptide of 11 residues that destroys β-folds (ΐΑβ1). Ίη νίνο studies have shown that a peptide carrying a natural sequence is rapidly degraded in rat plasma. Indeed, approximately 90% of ΐΑβ1 was degraded within a few minutes after intravenous administration. On the contrary, the ΐΑβ1 derivative, containing all the residues in the Ό-form, was practically not destroyed in the plasma within 15 minutes after the injection. To carry out detection, the peptide was subjected to radioactive iodination using standard methods. The stability of the peptides was evaluated after iv administration of boluses to rats by precipitation with trichloroacetic acid. Quantification of intact peptides was also carried out using paper chromatography. Thus, the ΐΑβ5 and 1PrP13 peptides (Figs. 3A and 3B, respectively) containing only Ό residues, as well as peptides containing Ό residues only at the и and C-terminal sites to prevent exopeptidase degradation, are included in the compounds of this invention. In addition to the above, the остаток residue was used after each proline amino acid, since it was reported that often endopeptidase cleavage occurs at the site located after the indicated residue under the action of an enzyme known as prolyl endopeptidase.

Alzheimer's Disease Inhibitors:

1st rgo pb pb azr 1st Rgo Pb Pb azr 1st Rgo pb Pb azr

Prion associated disease inhibitors:

azr a1a rgo a1a a1a rgo a1a d1u rgo a1a ua1 rgo ν31 azero A1a rgo A1a A1a rgo a1a s1u rgo a1a ua1 rgo νβΐ azr a1a rgo a1a a1a rgo a1a s1u rgo a1a aa

The amino acid designations starting with a lowercase letter refer to Ό-residues.

e) Cyclic peptides.

Conformationally strained cyclic peptides are better experimental drug samples than linear peptides due to their reduced conformational mobility and improved resistance to exopeptidase cleavage. Two alternative strategies were used to turn a linear sequence into a cyclic structure. One is the introduction of a cysteine residue to achieve cyclization through the formation of a disulfide bridge, and the other is a side chain attachment strategy involving head-to-tail cyclization on the resin. To avoid modifications to the peptide sequence, the latter approach is used. Peptides that destroy the β-folding structure contain ideal sequences to facilitate macrocyclization, since proline, thanks to its ability to initiate turns and loops, is an integral part of many naturally occurring or synthesized artificially cyclic peptides.

Alzheimer's Disease Inhibitors:

Prion associated disease inhibitors:

ί) Pseudopeptides.

Pseudopeptides or amide bond substitutes refer to peptides containing chemical modifications of some (or all) of the peptide bonds. Amide bond substitutions are usually denoted by keeping the amino acid designation in accordance with the side chain and describing the changes that occur among α-carbon atoms, using the nomenclature known as psi-bracket (pznbaske!).

For example, the term A1aΨ [SΗ2SΗ2] S1u refers to a fragment NΗ2SΗ _(CH3) CH2CH2CH2

CO ₂ N. Some substitutes for the amide bond are described below in table. 2.

Table 2. Some substitutes for the amide bond and their properties

Substitute The properties CH ₂ Short, mobile CH ₂ CH ₂ Mobile, hydrophobic sn = sn Tough, hydrophobic os Very tough CH ₂ CN Mobile, hydrophilic pine ₂ Mobile, hydrophilic sn ₂ s Mobile, hydrophobic CH ₂ ZO ₂ More rigid, hydrophilic cnso Hard, hydrophilic

Some of them are found in natural analogues of peptides (such as T [CHON], Ψ [Ο8ΝΗ], Ψ | ΟΌΟ |). while others were synthesized artificially. The introduction of amide bond substitutes not only reduces the degradation of peptides, but can also significantly change some of the biochemical properties of peptides, especially conformational mobility and hydrophobicity. It is possible that an increase in conformational mobility has a favorable effect on the binding of the inhibitor to the binding sites of Άβ and PrgP. On the other hand, since the interaction between amyloidogenic proteins and inhibitors seems to be largely dependent on hydrophobic interactions, it is possible that an increase in hydrophobicity by replacing the amide bond may increase the affinity and, therefore, the effectiveness of the inhibitors. In addition, increased hydrophobicity can also increase peptide transport across membranes and, thus, improve barrier permeability (blood-brain barrier and intestinal barrier). Therefore, for the synthesis of pseudopeptides, an amide bond substitution is used, such as Ψ [] ₂ ΟΗ ₂ ] and Ψ [ΟΗ ₂ δ], whereby mobility and hydrophobicity are increased. To prevent exoprotease degradation, the amide bonds located at the end of the peptide and after each of the proline residues are replaced, since it has been described that usually the endopeptidase cleavage sites under the action of an enzyme known as prolyl endopeptidase are located after this residue. Additional amide bonds that must be protected are determined by experimental studies, including analysis of the degradation of β-folding peptides in plasma and tissue.

Alzheimer's Disease Inhibitors:

Eif [CH ₂ CH _2] ΡΓθψ [0Η ₂ 0Η _2] RNU Ράθψ [CH ₂ CH _2] ei4 Asp _([2 CH 3] Rgof _[2 CH 3] Phe №ef _[2 CH 3] Asp

Prion associated disease inhibitors:

Azrf [CH ₂ CH ₂ ] A1a Rgof [CH ₂ CH ₂ ] A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u

Rgof [CH ₂ CH ₂ ] A1a Wa1 Rgof [CH ₂ CH ₂ ] Wa1

Azrf [CH ₂ 3] A1a Rgof [CH ₂ 3] A1a A1a Rgof [CH ₂ 3] A1a C1u Rgof [CH ₂ 3] A1a Va1 Rgof [CH ₂ 3] Va1

d) The combination of several modifications.

Considering the properties of peptide drugs that are commercially available or currently under development, it can be seen that most of the peptides that are resistant to proteolysis include a combination of several types of the above modifications. This conclusion makes sense in the light of data indicating the participation of many different enzymes in the degradation of peptides.

The following structures contain combinations of different types of chemical modifications:

Alzheimer's Disease Inhibitors:

As-Rhea Rhof [CH ₂ CH ₂ ] Rye Rye Azr-At

As-Rhei Rgof [CH ₂ 3] Rye Rye Azr-At (Me) Lye Rgof [CH ₂ CH ₂ ] Rye Rye Azr-At

1ey Rgof [CH ₂ CH ₂ ] Pbb Pbp azr

1ey Rgof [CH ₂ 3] Pbb Pbp azr

As-Ley A1 Rye Rye Azr-At (Me) Lye AFE Rye Rye Azr-At

Lye Rgos [CH ₂ CH ₂ ] Rye Rye Razr g - Lye Alb Rye Rye Azr - p - Lye Rgof [CH ₂ CH ₂ ] Rye Rye Azr · ^

As-ie rgo rie rie azr-at

As-Rhea Rhof [CH ₂ CH ₂ ] Pb-Pb-Azr-At

As-Rhee Rhof [CH ₂ 3] Pie Pie Azr-At

As-Rye Rhof [CH ₂ CH ₂ ] Pb (Me) Pb Azr-At

As-Lei Rgof [CH ₂ CH ₂ ] Pb (Me) Pb azr

As-ie Rgo Rie Rie Azr-At

As-Lei Rgo (Me) Rie Ree Azr-At

Ie Rhof [CH ₂ CH ₂ ] Pbpbp azr

1ee Rgo (Me)

As-Lei Al-Rie Rye Azr-At

Prion associated disease inhibitors:

As-Azr A1a Rgof [CH ₂ CH ₂ ] A1a A1a Rgof [CH ₂ CH ₂ ] A1 a C1u

Rgof [CH ₂ CH ₂ ] A1a Va1 Rgo Va1-At azr A.a Rgof [CH ₂ CH ₂ ] A1a A1a Rgof [CH ₂ CH ₂ ] A1 a C1u Rgof [CH ₂ CH ₂ ] A1a

Wa1 Rgo cha1

As-Azr A1a Rgof [CH ₂ 3] A1a A1a Rgof [CH ₂ 3] A1a C1u Rgof [CH ₂ Z] A1a

Ya1 Rgo Ya1-At azr A1a Rgof [CH ₂ 3] A1a A1a Rgof [CH ₂ 5] A1a C1u Rgof [CH ₂ 3] A1a Ya1

Rgo cha1

As-Azr A1a AFL A1a A1a AFL A1a C1u A1b A1a Ya1 Rgo Ya1-At

As-Azr A1a Rgof [CH ₂ CH ₂ ] A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u

Rgof [CH ₂ CH ₂ ] A1a Wa1 Rgo (Me) Wa1

As-Azr A1a rgo A1a A1a rgof [CH ₂ CH ₂ ] A1a C1u rgo A1a Va1 Rgo Va1Agg.

Azr A1a Rgof [CH ₂ CH ₂ ] A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u Rgof [CH ₂ CH ₂ ] A1a

Ua1 Pro (Me) Ua1 ASP A1a ΑίΡ A1a A1a Rgof [CH ₂ CH _2] A1a S1u pro L1a Ua1 Pro (Me) Ua1 ASP A1a ^ b A1a A1a Rgof [CH ₂ 3] A1a S1u pro A1a Ua1 His (Me) Ua1 ASP A1a Rgof [CH ₂ 3] A1a A1a Rgof [CH ₂ 3] A1a C1u Rgof [CH ₂ 8] A1a Уа1

Rgo (Me) Wa1

As-Azr A1a A1b A1.a A1a Rgof [CH ₂ CH ₂ ] A1a C1u Αίό A1a Va1 Rgo (Me) Va1 ι — Azr A1a rgo A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u rgo A1a Va1 Rgo Va1 —ι | —Azr A 1a ΑίΡ A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u ΑίΡ A1a (Me) Va1 Rgo Va1— |

As-Azr A1a Rgof [CH ₂ 3] A1a a1a Rgof [CH ₂ 3] A1a d1u Rgof [CH ₂ 5] A1a (Me) Wa! Rgo Ua1-At

As-Azr A1a ΑίΡ a1a A1a Rgof [СН ₂ СН ₂ ] А1а С1у рго А1а Уа1? Go (me) Уа1 azr AFa ΑίΡ А1.a A1a Ргоф [СН ₂ СН ₂ ] A1a С1у ΑίΡ а1а Уа1 Рго Уа1-Ат

Ac-Asp A1a pro A1a A1a Rgof [CH ₂ CH _2] A1a d! Y pro A1a (Me) Ua1 Pro Ua1-Am ASP A1a Rgof [CH ₂ CH _2] A1a A1a Rgof (CH ₂ CH ₂₎ A1a d! Y Rgof [СН ₂ СН ₂ ] А1а νβί Рго νθΐ

As-Azr A1a rgo A1a a! A ΑίΡ A1a d! U rgo A1a (Me) UAF Rgo Ua1-At

Azr A1a rgo A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u rgo A1a Wa1 Rgo Wa1

Azr L1a ΑίΡ A1a A1a Rgof [CH ₂ CH ₂ ] A1a C1u ΑίΡ A1a (Me) Wa! Rgo Wa1

There are numerous approaches to the construction and synthesis of peptidomimetics, as described in recent reviews of Wojcit Saye and Ggao Ofta e1 a1., Which are incorporated herein by reference.

The peptidomimetics below represent another aspect of the present invention.

Another approach to improving stability, which can also lead to the formation of active compounds upon oral administration, is to obtain a peptidomimetic. A peptidomimetic is a molecule that mimics the biological activity of peptides, but is not a peptide by chemical nature. The term peptidomimetic is sometimes used to describe molecules that are partially peptides in nature, such as pseudopeptides, semi-peptides or peptoids, but the strict definition and definition used in this application refers to an organic molecule that does not contain any peptide bonds at all. Peptidomimetics are not derivatives of the parent peptide, but are preferably chemically synthesized without trying to mimic the structural and functional properties of the peptide. For the rational construction of peptidomimetics, a sufficient level of knowledge of the pharmacophore groups responsible for activity and detailed structural information about the peptide are necessary. The goal is to recreate the spatial position of pharmacologically active groups using the organic matrix that supports them. Matrix selection is important, and size and mobility should be considered based on the conformational model of the peptide.

Peptidomimetics designed to mimic the properties of a peptide that destroys β folds

For the rational construction of peptidomimetics, a sufficient level of knowledge of the chemical groups responsible for the activity and detailed structural information about the peptide are necessary. The goal is to recreate the position of pharmacologically active groups using the organic matrix that supports them. Matrix selection is important, and size and mobility should be considered based on the conformational model of the peptide. As a result of the study of the activity of various sequences that destroy β folds carrying single amino acid substitutions, residues that play a key role in inhibition were determined. In addition, either a three-dimensional structure of peptides that destroy β-folds in Alzheimer's disease or a disease associated with prions was simulated or experimentally established (Figs. 3A and 3B). An inhibitor of Av fibrillogenesis containing 5 residues was modeled on the basis of the principle of minimum energy and the Moyer Sag1o method using the 1CM computer program. The structure of the 13-residual conformational change inhibitor in prion proteins was experimentally calculated using 2I-NMR.

Alzheimer's Disease Inhibitors

The latter inhibitor (PM1PgP5) is a shorter and easier to synthesize version that contains chemically active groups and is an analogue of a peptide of 5 residues that destroys β folds in prion disease.

As a method of preventing or treating a disorder or disease associated with amyloid or amyloid-like deposits, or their pathological precursors enriched with a β-folded structure, the compound of the present invention is administered in an effective amount to a subject in need thereof, where the subject may be a human or animal. Similarly, a method for diagnosing such disorders or diseases also includes administering the engineered compound in an amount sufficient to visualize its binding to fibril deposits or their precursors using well-known imaging methods.

As used herein, the term prophylaxis of a condition such as Alzheimer's disease or other amyloidosis disorders in a subject includes administration of a compound of the present invention prior to the clinical onset of the disease. Treatment includes administering a protective compound after the clinical onset of the disease. For example, the successful administration of a compound of the present invention after the development of a disorder or disease includes treatment of the disease. The present invention can be used to treat humans as well as veterinary animals.

The compound of the present invention may be administered by any method that achieves the intended purpose, preferably orally. For example, administration can be carried out by a number of different parenteral routes, including, without limitation, the subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, intracerebral, intranasal, oral, percutaneous or buccal routes. Parenteral administration may be by injection of a bolus or by gradual infusion over a period of time.

A typical regimen for the prevention, suppression, or treatment of a condition associated with amyloid or amyloid-like deposits includes: or (1) administering an effective amount of the compound in one or two doses containing a high concentration in the range of 0.5 to 10 mg, more preferably , from 0.5 to 5 mg, or (2) the introduction of an effective amount of the compound in repeated doses containing lower concentrations in the range of 1010000 μg, more preferably 50-500 μg, over a period of time from several months up to several years.

It is clear that the dose administered depends on the age, gender, state of health and weight of the recipient, the type of concomitant treatment, if any, the frequency of treatment and the nature of the desired effect. The total dose required for each treatment can be administered in repeated doses or in a single dose. An effective amount is understood to mean a concentration of a compound that is capable of slowing down or inhibiting the formation of amyloid or amyloid-like deposits, or their pathological β-folded precursors, or dissolving previously formed fibril deposits. Such concentrations may be determined by those skilled in the art in a routine manner. Those skilled in the art will also appreciate that the dosage may depend on the stability of the administered compound. For a less stable compound, multiple doses may be required.

Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions, which may contain auxiliary agents or excipients known in the art. Pharmaceutical compositions such as tablets and capsules may also be prepared according to conventional methods.

Pharmaceutical compositions comprising a compound of the present invention include all compositions in which the compound is contained in an amount effective to achieve the intended purpose. In addition, the pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers, including excipients and excipients, which facilitate the processing of the active compounds in preparations that can be used pharmaceutically. Suitable pharmaceutically acceptable media are well known in the art and are described, for example, in Oeppago, Ebn δο, Eb., Kesh1pd1op'5 Ryagtaseibs1 8c1epse5, 18 ^4b Ebyup 1990, Mask Ryyppd Co., Ea§1op, RA, standard guide to this field. Pharmaceutically acceptable environments can be selected in the working order in accordance with the method of administration, solubility and stability of the compounds. For example, compositions for intravenous administration may include sterile aqueous solutions, which may also contain buffers, diluents, and other suitable additives.

Suitable compositions for parenteral administration include aqueous solutions of the active compounds in a water-soluble form, for example, in the form of water-soluble salts. In addition, a suspension of the active compound in the form of appropriate oily injection suspensions may be administered. Suitable lipophilic solvents or media include fatty oils, for example sesame oil, or synthetic fatty acid esters, for example ethyl oleate or triglycerides. Aqueous injection suspensions, which may contain substances that increase the viscosity of the suspension, include, for example, sodium carboxymethyl cellulose, sorbitol and / or dextran. Optionally, the suspension may also contain stabilizers.

Disorders or diseases associated with abnormal folding of proteins in amyloid or amyloid-like deposits, or in pathological precursors of such deposits enriched with β-folds, to be treated or prevented by administering a pharmaceutical composition of the invention include, but are not limited to, Alzheimer's disease, RAB, syndrome Down, other amyloidosis-related disorders associated with prion diseases, such as Kuru, Creutzfeldt-Jakob disease (STO), Gerstman-Strausslet-Sheinker syndrome (Seg81tapp- 1gai551e18sye1pkeg 8upbgote (688)), human neurodegenerative diseases associated with prions and prion-related animal diseases such as scrapie, spongiform encephalopathy, transmissible mink encephalopathy and chronic wasting disease of mule deer and elk.

Examples

One of the main disadvantages of the use of peptides as medicines is their rapid proteolytic degradation in biological fluids and tissues. In the experiments of shuigo ίΑβ5 (the last BRBBO, also referred to in this description as

Rgoryeuryeacr) is destroyed very quickly after incubation of ίη νίΐτο with fresh human plasma. As shown in FIG. 4A, fifty percent of the β5 peptide in the presence of plasma disappears within approximately 5 minutes. Since it is impossible to identify any metabolic fragments resulting from proteolytic cleavage, it is likely that degradation mainly occurs under the influence of nonspecific exopeptidases. This conclusion is confirmed by the fact that the protection of the amino and carboxy-terminal groups of the peptide by acetylation and amidation, respectively (with the formation of Αο-ίΑβ5-Αιη. Also referred to in this description as Ac-Liérgo Riere Akr-At) significantly increases the stability of the ίη νίΐτο peptide. As shown in FIG. 4B, the modified protected-end peptide of the present invention (Ac-1A β5At) remains stable in human plasma for a period exceeding 24 hours. (The modified peptide is also slowly metabolized by ίη νίΐτο in human and rat liver microsomes, in which after one hour of incubation with 37 ° C 81.5% and 76.3% of the peptide remains intact in the human and rat tissue homogenate, respectively.)

Additional studies of ίη νίΐτο showed that Ac-1Av5-At has the same activity in inhibiting the formation of amyloid as ί A [35 (see Fig. 5A), and the effect has the same dose dependence as the activity of the unmodified peptide, as shown in figv and 5C. Returning to FIG. 5A, it can be seen that the modification of the конца-terminus with Boc also allows one to retain the ίη νίΐτο activity exhibited by 1Ab5, while some unrelated peptides (CP1: UNU8EEOOTER, CP2: BUTAUAUEB, CP10: 18EUKMOEE) or short fragments such as Λβ 18-21, Av 1-16) at the same concentration do not affect fibrillogenesis or slightly increase the formation of amyloid by inclusion in fibrils.

In order to evaluate the effect of Ac-ί A [35At ίη νίνο, a rat model is used in which amyloidosis is induced by intracerebral injection of non-aggregated Av-42. After some time, the peptide aggregates in the rat brain, leading to the formation of a single amyloid-like deposit at the injection site. These lesions have the same staining (birefringence with Congo red and thioflavin 8 binding) and light transmitting (fibrillar structure by electron microscopy) properties as amyloid plaques in Alzheimer's disease and cause some brain damage similar to that observed in AD, including extensive compression of neurons, astrocytosis and activation of microglia. Using this model, the inventors have shown that a joint injection of unprotected ί A [35 and Av 1-42 causes a 50% inhibition of the formation of amyloid plaques, and i.c. (intracerebral) injection of ί A [35 to animals already containing amyloid plaques causes the dissolution of 67% of previously formed deposits (8φιΐΓά55οη, E.M., Retinsche, V., 8οΐο, C, XVί5shekhhkkg T. & Επιηφοί'Κ, B . (2000) Ιη νίνο άί5355et1u οί atu ΙοίΠ-β ώ ^ ροδίΐδ ίη ha1 Part 1. 1.ιιτοραΐΐι. Exp. ΝοιιγοΙ. 59: 11-17). In previous experiments, the unprotected peptide was introduced directly into the area of the brain in which the amyloid was located. In the present experiment, the amyloid 5 peptide is introduced into the amygdala of rats. After 7 days, the time required for complete completion of the formation of amyloid deposits, 100 μl of a solution containing 13 mg / ml Αc- ^ Αβ5-Αt was poured over three weeks using an ABKHET infusion pump connected to the lateral ventricle. Animals were sacrificed, and the brain was analyzed for the presence of amyloid deposits by the immunohistochemical method. In this model, a compact amyloid plaque was obtained in the area (amygdala), in which there was a solution containing Aβ1-42, as well as several smaller amyloid deposits were observed along the cannula path in areas closer to the ventricle (Fig. 6, left panel ) The results show that peptide infusion causes dissolution of 30% of the previously formed amyloid plaque and 83% of deposits located near the ventricle (Fig. 6).

Experimental Methods Peptide Stability Assays ίη νίΐτο

Peptides are obtained as a solution in water with a concentration of 1 μg / ml. 20 μl of the peptide solution is diluted in 80 μl of fresh human plasma. The solution was incubated at 37 ° C for different periods of time, and the reaction was stopped by adding a complete set of protease inhibitors. Most plasma proteins (but not peptides) are precipitated with cold methanol (mixture / MeOH 4/5, v / v) for one hour at -20 ° C. Precipitated proteins are precipitated by centrifugation (10,000 d 10 min, 4 ° C). The peptide-containing supernatant was concentrated 5-fold under vacuum and separated by reverse phase HPLC. The peak region corresponding to the intact peptide is measured and compared to the peak area corresponding to the equivalent sample incubated without plasma.

Activity Analyzes ίη νίΐτο

Amyloid formation is quantified by fluorescence emission of thioflavin T (T11T) associated with amyloid fibrils. Aliquots of a solution of β with a concentration of 0.5 mg / ml, prepared in 0.1 M TP5, pH

7.4, incubated for 7 days at 37 ° C in the absence or presence of various concentrations of β5 and its derivatives. At the end of the incubation period, 50 mM glycine, pH 9.2, and 2 μM Tc are added in a final volume of 2 ml. Fluorescence is measured at an excitation wavelength of 435 nm and an emission wavelength of 485 nm on a Retksh Е1teg fluorescence spectrometer, model L850B.

Ιη νίνο studies using an animal model of cerebral AR deposition

At the time of receipt, the age of male Nkkeg-344 rats is 3-4 months, and the weight is 250-300 g. Animals are placed in cages, 2 each, kept in a 12-hour cycle of alternating light and dark with access to food and water, and Nyyit and accustom to new medium for 2–3 weeks before surgery. Surgery is performed under anesthesia with sodium pentobarbital (50 mg / kg, ip). Atropine sulfate (0.4 mg / kg) and ampicillin sodium salt (50 mg / kg) are subcutaneously administered to anesthetized animals. Av 1-42 is dissolved in dimethyl sulfoxide (ΌΜ8Ο) and then diluted with water to obtain 16.7% ΌΜ8Ο. The animal receives a bilateral injection of 5.0 nmol Av 1-42 into each amygdala, which is carried out using the stereotactic instrument Kor £ with a cutter blade set 3.3 mm below the center line. Injection coordinates, measured from the crown of the head and the surface of the skull (AP-3.0, M ± 4.6 BU -8.8), were empirically determined based on the atlas of Paxinos (Rahshok) and Watson (\ Va1kop). A volume of 3.0 μl was injected over 6 minutes (flow rate 0.5 μl / min) using a CMA / 100 micro-syringe pump. The cannula was left ίη κίΐιι for 2 minutes after the injection, then it was removed 0.2 mm and left for 3 minutes, and after 5 minutes the cannula was slowly removed. After the surgery, the animals are placed on a heated pillow until their installation reflex is restored. To evaluate the effect of Λο-β5-Λιη, the animals undergo a second surgery one week after the first, in which the infusion pump ΑΕΖΕΤ is connected to the ventricle of the brain, following the manufacturer's instructions. A total amount of 1.3 mg of the peptide in 100 μl of PB8 / 10% ΌΜ8Ο is delivered to the lateral ventricle within 3 weeks. After this period of time, the animals were euthanized with an excess dose of sodium pentobarbital (150 mg / kg, ip) infused through the aorta. For histological analysis, sequential coronary segments (40 μm) of the brain are excised, placed in an ethylene glycol cryoprotectant and stored at -20 ° C until staining. Tissue segments are stained with antibodies against Av 1-42, as described in 8 °! O, C, 81 digkop, E., Motesh, L., Kitag, Β.Α., SakRto. EAT. apy Etapdupe, V. (1998) β-kkey! Геgeakeg rerrejek ίηΙιίΝι DtShodepeak ίη and that! This is a toyo о ат у 1 1 о й ок 1 1 1::: kirksa Roth, ог ог Л х к ек ек ек п '' к к

Shetar. Eeyore! 4: 822-826. A visual analysis system is used to determine the size of amyloid deposits. The data is analyzed using a two-way ΑΝΟνΑ followed by testing with IE ^ that ^^ and AI with many ranges for rock! braid comparisons. The total amount of deposits in the brain is analyzed using an unpaired! -Test, with double sampling.

Having now a complete description of the invention, those skilled in the art can appreciate that such experiments can be carried out over a wide range of equivalent parameters, concentrations and conditions without deviating from the essence and scope of the invention and without proper experiments.

Since the present invention has been described by way of specific embodiments thereof, it should be understood that it may be subjected to further modifications. This application is intended to cover any variations, applications or adaptations of the inventions, subject to the general principles of the present invention and includes deviations from this description, such as arising within the framework of a known or habitual practice in the field to which the invention relates, and such as which can be applied to the essential features described above, as follows from the scope of the attached formula.

All references cited in this description, including journal articles or abstracts, published or relevant patent applications of the United States or other countries, or any other references, are fully incorporated into this description, including all data, tables, numbers and text provided in the cited references. Additionally, the full content of the cited references in the scope of the references cited in this description is also fully incorporated by reference.

References to known stages of the method, conventional stages of methods, known or conventional methods in no way allow the assumption that any aspect, description or embodiment of the present invention is disclosed, implied or implied in the related field.

The present description of specific embodiments thus reflects the general nature of the invention that other persons, having applied knowledge of skills in this field (including the contents of the references cited here), can easily modify and / or adapt these specific embodiments for various applications without undue experimentation and departure from general concept of the present invention. Therefore, it is understood that such adaptations and modifications are within the meaning and range of equivalents of the disclosed embodiments, based on the guidance and guidance provided herein. It should be understood that the phraseology or terminology in this document serves the purpose of description, and not limitation, so that the terminology or phraseology of the present description should be interpreted by a qualified specialist in this field in light of the guidance and guidance presented in this document, combined with the knowledge of an ordinary specialist in this area.

List of sequences <110> Ahopuh, 1ps.

<120> Peptide analogues and mimetics suitable for the use of ίη νίνο in the treatment of diseases associated with abnormal folding of ebc proteins.

<130> Ahopuh-YUi-4 <140> PCT / C300 / 30416 <141> 2000-11-04 <160> 4 <170> RakepMp Ueg. 2.0 <210> 1 <211> 5 <212> PKT <213> Artificial sequence <220>

<223> Description of the artificial sequence: peptide obtained by chemical synthesis <400> 1

Bei Rgo Rye Rye Azr <210> 2 <211> 13 <212> RKT <213> Artificial sequence <220>

<223> Description of the artificial sequence: peptide obtained by chemical synthesis <400> 2

Azr A1a Rgo A1a A1a Rgo A1a O1u Rgo A1a Wa1 Rgo Wa1

10 <210> 3 <211> 5 <212> PKT <213> Artificial sequence <220>

<221> MOE_KEZ <222> (2) <223> Haa represents A1b <220>

<223> Description of the artificial sequence: peptide obtained by chemical synthesis <400> 3

Ley Haa Rye Rye Azr

5 <210> 4 <211> 13 <212> PKT <213> Artificial sequence <220>

<221> MOE_KEZ <222> (3) <223> Haa represents ΑίΡ <220>

<221> MOO__KEZ <222> (6) <223> Haa represents ΑίΡ <220>

<221> MOR_KEZ <222> (9) <223> Haa represents ΑίΡ <220>

<221> MOO_KEZ <222> (12) <223> Haa represents ΑίΡ <220>

<223> Description of the artificial sequence: peptide obtained by chemical synthesis <400> 4

Azr A1a Haa A1a A1a Haa A1a <31u Haa A1a Wa1 Haa Wa1

10

Claims (23)

CLAIM 1. An inhibitory peptide capable of inhibiting the formation of a β-folded structure in an amyloid β-peptide, which is an analogue of the inhibitor peptide of Alzheimer's disease пептиβ5, consisting of 5 residues (8EO GO N0: 1 Bsi-Pro-PHC-RHC-A ^ p). constructed by chemical modification 8E0 GO N0: 1, wherein said chemical modification is achieved using a method selected from the group consisting of altering the Ν- and C-termini of said inhibitory Alzheimer's disease peptide ίΑβ5; replacing at least one residue of the indicated Alzheimer's inhibitory peptide βίΑβ5 with α-aminoisobutyric acid (A1B); methylation of the α-carbon atom of at least one residue of the indicated Alzheimer's disease inhibitor peptide ίΑβ5; replacing at least one L-enantiomeric residue of the indicated Alzheimer's disease inhibitor peptide ίΑβ5 with the Ό-enantiomeric residue; performing head-to-tail cyclization of the indicated Alzheimer's disease inhibitory peptide ίΑβ5; their combinations. 2. The inhibitory peptide according to claim 1, characterized in that said change in Ν- and C-ends of said inhibitor peptide of Alzheimer's disease ίΑβ5 is achieved using a method selected from acetylation, amidation, deamination, alcoholization and their combinations. 3. The inhibitory peptide according to claim 2, characterized in that said inhibitory peptide is selected from the group consisting of as-Lei Pro-Pere PeArAr-at, des-Lei Pro-Peu-Pee-Bee-Acru, as-Bei Proe-Bee a1c and des-lei Rgo Rye Pye Azr-a1c. 4. The inhibitory peptide according to claim 3, characterized in that said inhibitory peptide is ac-lei Pro-Pyrbe Pee ΑκρAT. 5. The inhibitor peptide according to claim 1, characterized in that said inhibitory peptide is selected from the group consisting of Bei a1b Pee Pee (Me) Bee Rgo Rye Azr; Pye Azr; Bei Rgo (Me) Pye Pye Azr, Bei Rgo Pye (Me) Pye (Me) Lei Rgo (Me) Pye Azr; (Me) Pye Azr, 1e rgo pyr pye azr, Bei Pro Pye Pye Azr, Pbe Leif {CH ₂ CH ₂ ] Proth [CH ₂ CH ₂ ] Pbe Péf [CH ₂ CH ₂ ] Azr; Beif [CH ₂ 5] Pgοψ [CH ₂ 5] Pye Prief [CH ₂ 3] Azr; Al-Lei Rgoff [CH ₂ CH ₂ ] Pée Pée Azr-At; Al-Lei Rougf [CH ₂ 3] Pbe Peh Al Az-At; (Me) Lei Rgoff [CH ₂ CH ₂ ] Pye Pye Azr-At; Bei Pro Pye ar, Lei Rgoff [CH ₂ CH ₂ ] Pbe Pie Azr; Béy Ρτοψ [CH ₂ 3] Pée Péé al; Al-Bey ABE Pere Pere Azr-At; (Me) Lei ABe Pere Pere Azr-At; Lei Rgoff [CH ₂ CH ₂ ] Pbe Pie Azr; | —Lee ABL Pye Pye Azr-η residue, performing head-tail tail cyclization of the indicated prion inhibitory peptide 1Pr13, replacing amide bonds in the indicated prion inhibitory peptide 1RgP13 with an amide bond substitute and their combinations. 6. The inhibitor peptide according to claim 1, characterized in that said inhibitory peptide is characterized by the following structure: 7. An inhibitory peptide capable of inhibiting conformational changes in the prion protein of RgP associated with amyloidosis, which is an analogue of the prion inhibitory peptide 1PrP13, consisting of 13 residues (8EO GO N0: 2 ρκρ Α1; · ιPgo Α1; · ι 1; · ι Rgo Α1; · ι О1у Rgo Α1; · ι Уа1 Rgo Уа1), constructed by the chemical modification 8E0 GO N0: 2, while the indicated chemical modification is achieved using a method selected from the group consisting of changing Ν- and C-ends of the specified prion inhibitor peptide 1PrP13; replacing at least one residue of the indicated prion inhibitory peptide 1Pr13 with α-aminoisobutyric acid (ΑίΒ); methylation of the α-carbon atom of at least one residue of the indicated prion inhibitory peptide gRgP13; substitution of at least one L-enantiomeric residue of the indicated prion inhibitory peptide gRgP13 with Ό-enantiomeric Azrf [СН ₂ СН ₂ ] АБа Ргоф [СН ₂ СН ₂ ] АБа АБа Ргоф [СН ₂ СН ₂ ] АБа СЬ Роф [СН ₂ СН ₂ ) АБа УаЬ Ргоф [СН ₂ СН ₂ ] Va; Azrf (СН ₂ 3] АБа Ргоф [СН ₂ 3] А1а АБа Ргоф [СН ₂ 3] АБа СЬу Ρτοψ [СН ₂ 3] АБа Wah Ruhf [CH ₂ 3] Wah; Al-Azr ABa Roug [CH ₂ CH ₂ ] ABA AbA Roug [CH ₂ CH ₂ ] ABA Cy F Rouf (CH ₂ CH ₂ ] ABA Wa Pogo Wo-At; azr aba rugof (СН ₂ СН ₂ ) aba aba rug [СН ₂ СН ₂ ] aba cb rugоg [СН ₂ СН ₂ ] aba Wah Rgo We1; Al-Azr Aba Ruhoff [CH ₂ 5] Aba Aba Rukhgof (CH ₂ 3) Aba Chu Ruhoff [CH ₂ 3] Aba Wa Rgo Wa-At; azr aba rugof [CH ₂ 3] aba aba rugof [CH ₂ 3] aba Chu rugof [CH ₂ 3] aba Wa Pogo wa; As-Azr ABA ABB ABA ABA ABB ABA Cyb ABE ABA WA Pro Rgo Wo-At; Al-Azr ABa PiFo [CH ₂ CH ₂ ] A1A Aba PiFo [CH ₂ CH ₂ ] ABA CyU Pyo [CH ₂ CH ₂ ] ABa Wa Pro (Me) UA; As-Azr ABa pro GAO ABA ABA Pragof [CH ₂ CH ₂ ] ABA Cb o pro GODAA Va Pro Pro; azr aba rugof (СН ₂ СН ₂ ) aba aba rug [СН ₂ СН ₂ ) А1а СЬу rug [СН ₂ СН ₂ ] aba Wah Rgo (Me) Wah; azr aba abh aba aba rugof [СН ₂ СН ₂ ] aba cba rgo aba wa pgo (me) wa; azr aba abh aba aba rugof [CH ₂ 3] aba syb pro abo aba ba pro (me) wo; Asr Aba Roug (CH ₂ 3) Aba Aba Roug [CH ₂ 3] Aba Chu Roug [CH ₂ 3] Aba Wa Pro (Me) Wa; Al-Azr Aba Abh Aba Aba Ruhoff (CH ₂ CH ₂ ] Aba Su Fu Abu Aba Wa Proe (Me) Wa; Al-Azr Aba Roug (CH ₂ 3] Aba Aba Prog [CH ₂ 3] Aba Dob Prog [CH ₂ 3] Aba (Me) Wa Pro Roh-At; Al-Azr Aba Abh Aba Aba Rufo [CH ₂ CH ₂ ) Aba Chu Pgo Aba Wa Pro (Me) Wa; azr aba abh aba aba rugof [CH ₂ CH ₂ ] A1a cb ab aba aba proggo wa-At; As-Azr ABa rgo ABA ABa PiFo [СН2СН2] ABA DoBu ProVO ABa (Me) Dia1 ProPgo Wa-At; AZA Aba Profof [CH ₂ CH ₂ ] Aba Aba Prog (CH ₂ CH ₂ ) Aba dy Proh (CH ₂ CH ₂ ) ABa Wa Prog; Al-Azr Aba pro, Aba Aba Abu Aba Du, Pro Abo (Me) Wa Proe Wa-At; Azr Aba pro Got Aba Aba Roug [CH ₂ CH ₂ ] A1a Chu pro Got Aba Wa Gly; Azr Aba Ab Aba Aba Aba Roug [СН ₂ СН ₂ ] Aba Sy Be Ab Aba (Me) Wa Proe; and g - Azr Aba Pro Rgo Aba Aba Pro Rgo Aba Chu Pro Rgo Aba WA Pro Rgo Wa-v 8. The inhibitor peptide according to claim 7, characterized in that said change in Ν- and C-ends of said 1PgP13 prion inhibitory peptide is achieved using a method selected from acetylation, amidation, deamination, alcoholization, and combinations thereof. | —Lee Rgoff [СН2СН2] Pye Pye Azr Al-bei Al-bei Al-bei As-Lei rgo Pye Pere Azr-At; Ψτοψ [СН2СН2] Pye pye Azr-At; Ruhf (CH ₂ 3] Pyrbere Be Azr-At; Rgoff [CH ₂ CH ₂ ] Pbe (Me) Pbe Azr-At; 9. The inhibitor peptide of claim 8, characterized in that said inhibitory peptide is selected from the group consisting of ac-Azr ABa Pro-A1A A1a Pro-ABA C1O Pro-A1A Ua Pro Pgo Ua-at, c-ez-Azr Aba PAO ABA Aba Pro ABA Chu Rgo ABA WA Rgo WA-AT, Al-Azr ABA Rgo ABA ABA Rgo ABA Chu Rgo ABA WA Rogo UA-Ac and des-Azr ABA Rgo ABA ABA Rgo ABA Cy Rogo ABA Circued 10. The inhibitor peptide according to claim 7, characterized in that said inhibitory peptide is selected from the group consisting of Al-bei Rgoff [CH ₂ CH ₂ ] Pbe (Me) Pbe azr; Proe roe roe Al-bei Rgo (Me) Pye Azr-At; Peh Azr-At; Pere Azr; Bei Rgo (Me) Pere Pere Azr; Azr Aba Ab Aba Aba Ab Aba Aba A1b Aba Si Ab Aba Wah ab wah Azr Aba Pro Aba Aba Pro Aba Si Pro Aba (Me) ¹ wah Pro Wah; Azr Aba Pro Aba Aba Pro Aba Si Pro Aba Wah Pro (Me) Wah; Azr Aba Pro Aba Aba Pro Aba Si Pro Aba (Me) ι wah Pro (Me) Wa; azr aba pro aba aba pro a1a doo pro aba wah! pro Wah; azr Aba Pro Aba Aba Pro Aba Si Pro Aba Wah Pro wah azr Aba Pro aba Aba Pro aba Si Pro aba Wah Pro wah; Lei Rgoff [CH ₂ CH ₂ ] R'e 11. The inhibitor peptide according to claim 7, characterized in that said inhibitory peptide is characterized by the following structure: PM1RGR13 12. A method of reducing the formation of amyloid or amyloid-like deposits, including the abnormal laying of amyloid β-peptide in the β-folded structure, or reducing the amount of the specified amyloid β-peptide that has already been formed into a beta-folded structure, including contacting the specified amyloid βpeptide, either before or after its abnormal stacking in the β-folded structure with an effective amount of the peptide according to claim 1. 13. A method of reducing the formation of amyloid or amyloid-like deposits, including conformational changes in the prion protein Pg, or reducing the amount of the specified prion protein Pg, which has already formed into amyloid or amyloid-like deposits, including contacting the specified prion protein Pg, either before or after the specified conformational changes , leading to the formation of amyloid deposits with an effective amount of the peptide according to claim 7. 14. A method for reducing the formation of amyloid or amyloid-like deposits by administering an inhibitory peptide selected 15. Pharmaceutical composition comprising inhibitory peptide inhibiting the formation of β-folded structure in the amyloid β-peptide, and a pharmaceutically acceptable carrier or excipient, with the specified inhibitory peptide is an analogue of the inhibitory peptide of the disease Alzheimer ίΑβ5, consisting of 5 residues (8E0 GO N0: 1 Euei-Pro-Ryye-Ryye-Acre), designed by chemical modification 8E0 GO N0: 1, while the specified chemical modification is achieved using a method selected from the group consisting of changes in the Ν and C-termini of said inhibitor peptide of Alzheimer's disease ίΑβ5; replacing at least one residue of the indicated Alzheimer's inhibitory peptide βίΑβ5 with α-aminoisobutyric acid (A1B); methylation of the α-carbon atom of at least one residue of the indicated Alzheimer's disease inhibitory peptide ίΑβ5; replacing at least one L-enantiomeric residue of the indicated Alzheimer's disease inhibitor peptide ίΑβ5 with the Ό-enantiomeric residue; performing head-to-tail cyclization of the indicated Alzheimer's disease inhibitory peptide ίΑβ5; their combinations. 16. The pharmaceutical composition according to claim 15, characterized in that said inhibitory peptide is selected from the group consisting of as-Loe Pro-Pere Pere Azr-at, Joez-Loe Pro-Pere Pere Bere-Ache, as-Bey-Lo Pgo Pehroe Asra- a1c and c! ez-bei rgo pye pye azr-a1c. 17. The pharmaceutical composition according to p. 15, characterized in that the inhibitory peptide is an ac-lei Pro-Pyrbe Pee Ακρ-at. 18. The pharmaceutical composition according to p. 15, characterized in that the inhibitory peptide is selected from the group consisting of Bei A1b Pye Pye Azr; (Me) bei Pro Pye Pye Azr; Bei Pro (Me) Pye Pye Azr, Bei Pro Pye (Me) Pye Azr; (Me) bei Pro (Me) Pye (Me) Pye Azr, 1s pro rbe rbe azr, 1st Rgo Pye Pie Azr, 1s Rgo Ryu Pye Azr, Leite] / [CH ₂ CH ₂ ] PGO1INSH ₂ CH ₂ ] Pbé Peref [CH ₂ CH ₂ ] Azr; Lesh | r (CH ₂ 3] ΡΓΟψ [CH ₂ 3] Pbe ΡΗ β ψ [CH ₂ 3] Azr; Al-Lei Pro1] g (CH ₂ CH ₂ ] Pye Pere Azr-At; Al-Lei Ρτοψ [CH ₂ 5] Pye-Pye-Azr-At; (Me) Lei ψτοψ [CH ₂ CH ₂ ] Pée Pye Azr-At; 1s Rgoff [CH ₂ CH ₂ ] Pbe Pie Azr; 1 Ρτοψ [0Η ₂ 5] Pye Le Pie azr; Al-Bey Aly Pe'e Pe'e Azr-At; (Me) Bei Ah Pee Beh-Al-At; Béy Ρτοψ [CH ₂ CH ₂ ] Pée Péé al; I — lei Ach Pee-Pye Azr - “| —Bei ψτοψ [CH ₂ CH ₂ ] Pee-Pye Azr— | As-Lei rgo Pye Pere Azr-At; Al-Bey Ρτοψ [CH ₂ CH ₂ ] Pere Pere Azr-At; Al-Lei Ρτοψ [CH ₂ 3] Pierre Peh AzAr-At; Al-Lei Ρτοψ [CH ₂ CH ₂ ] Pye (Me) Pye Az-At; Ac-lei Pro1 | g [CH ₂ CH ₂ ] Pbe (Me) Pbe al; As-Lei Proere Beyr Azr-At; Al-Lei Rgo (Me) Pyré Pere Azr-At; 1e Ρτοψ [CH ₂ CH ₂ ] Pere PeEr azr; 1s Pro (Me) Pee Pere Azr; As-Lei Aib Pere Peh Azr-At; and | - Bei Rgo Rye Pye Azr - | 19. The pharmaceutical composition according to claim 15, characterized in that said inhibitor 27 peptide is characterized by the following structure: ⁿ GC ^ CH ₃ 9 soon ην ^ - '' Hun CH _E 0 ΡΜ1Λβ5 20. A pharmaceutical composition comprising an inhibitory peptide that inhibits conformational changes in the prion protein of PgP associated with amyloidosis, and a pharmaceutically acceptable carrier or excipient, with the indicated inhibitory peptide being an analog of the prion inhibitory peptide 1PgP13 consisting of 13 residues (δΕΟ ΙΌ N0: 2 Acre A1a Pro Ago A1a Pro Ago C1o Pro Ago Aa1 Pro Ugo1), constructed by chemical modification δΕΟ ΙΌ N0: 2, with the specified chemical modification achieved using a method selected from uppy consisting of Ν- changes and C-termini of said prion inhibitory peptide 1RgR13; replacing at least one residue of the indicated prion inhibitory peptide 1Pr13 with β-aminoisobutyric acid (A1B); methylation of an aaa-carbon atom of at least one residue of said prion inhibitory peptide 1Pr13; replacing at least one L-enantiomeric residue of the indicated prion inhibitory peptide 1РгР13 with the анти-enantiomeric residue; cyclizing head-to-tail of the indicated prion inhibitory peptide 1РРР13; 21. The pharmaceutical composition according to claim 20, characterized in that said inhibitory peptide is selected from the group consisting of ac-Azr A1a Pro Ago A1a Pro Ago A1a Pi Pro A1a \ / a1 Pro Aco A1A A1A A1a Pro А1а С1у Рго А1а Уа1 Рго Уа1-ата, as-Azr А1а Рго А1а А1а Рго А1а 01у Рго А1а Уа1 Рго Уа1-а1с and <un-Azr А1а Рго А1а Ааа Рго А1а <31у Рго А1а А1а Ро Ара Ро Ара Аго 22. The pharmaceutical composition according to claim 20, characterized in that said inhibitory peptide is selected from the group consisting of Azr A1a ΑίΡ A1a A1a ΑίΡ A1a A1a ΑίΡ A1a S1u A1P A1A UA1 ΑίΡ UA1 Azr A1a Pro A1a A1a Pro A1a S1u Pro A1a (Me: ΐ UA1 ProWa1; Azr A1a Pro A1a A1a Pro A1a S1u Pro A1a UA1 Pro (Me) UA1; Azr A1a Pro A1a A1a Pro A1a S1u Pro A1a (Me] > Va1 Pro (Me) Wa1; azr a1a pro and! a1a pro a1a d1u pro a1a νβΐ pro cha1; azr A1a Pro A1a A1a Pro A1a S1u Pro A1a UA1 Pro UA1; azr A1a Pro and! A1a Pro and! S1u Pro and! UA1 Pro UA1; Azrf (CH ₂ CH ₂ ) A1a Pink [SNgSNg] A1a A1a Pink [CH ₂ CH ₂ ] A1 A Cl Ргоф [СН ₂ СН ₂ ] А1а Уа1 Ргоф [СН ₂ СН ₂ ] Уа1; Azrf (СН ₂ 3] А1а Ргоф [СН ₂ 3] А1а А1а Ргоф [СН ₂ 3] А1а С1у Ргоф [СН ₂ 3] А1а Уа1 Pro1 | g (СН ₂ 3] Уа1; Ac-Azr A1a Profof (CH ₂ CH ₂ ] A1a A1a Prof [CH ₂ CH ₂ ] A1a Clou Prof [CH ₂ CH ₂ ] A1a UA1 Pro x / a1-At; Gap A1a PiFo [CH ₂ CH ₂ ) A1a A1A Piof (CH ₂ CH ₂ ) A1a Cyi Piogo [CH ₂ CH ₂ ] A1a Va1 Pro νβΐ; Ac-Azr A1a Proff [CH ₂ 3] A1a A1a Proff [SNGZ] A1a C1u Proff [CH ₂ 3] A1a \ 7-1 Rgo Wa-At; al papa A1a Prof [CH ₂ 3] A1a A1a Proff [CH ₂ 3] A1a C1u Prof [CH ₂ 3] A1a Ua1 Pro νβ; As-Azr A1a A1b A1a A1a A1b A1a C1u ΑίΡ A1a UA1 Rgo Wa1-At; Ac-Azr A1a Prof [CH ₂ CH ₂ ] A1a A1a Prof [CH ₂ CH ₂ ] A1a Clou Prof [CH ₂ CH ₂ ] A1a Ua1 Pro (Me) Ua1; Ac-Azr A1a rgo A1a A1a Profo [CH ₂ CH ₂ ] A1a C1u rgo A1a Ua1 Progo A1At; alp A1a PiFo [CH ₂ CH ₂ ] A1A A1A Pihof [CH ₂ CH ₂ ] A1A C1u Pigo [CH ₂ CH ₂ ] A1A UA1 Pro (Me) UA1; alp A1a A1b A1a A1a Prof [CH ₂ CH ₂ ] A1a C1u rgo A1a Vi1 Pi (Me) Dia1; alp A1a ΑίΡ A1a A1a Roug (СН ₂ 3] A1a С1у рго А1а Уа1 Рго (Ме) Уа1; ASP A1a Rgof [CH ₂ 3) A1a A1a Rgof _[CH2 8] A1a S1u Rgof [CH ₂ 3] νβΐ A1a Pro (Me) Ua1; Ac-Azr A1a ΑίΡ A1a A1a Roug [CH ₂ CH ₂ ] A1a C1u ΑίΡ A1a Vi1 Pro (Me) Dia1; 23. The pharmaceutical composition according to claim 20, characterized in that the inhibitory peptide is characterized by the following structure: