HK1236041A1 - Peptides - Google Patents

Description

Peptides

The present invention relates to peptides having dual BACE1 inhibitory properties, their manufacture, pharmaceutical compositions containing them and their use as therapeutically active substances.

Technical Field

The compounds of the present invention have Asp2(β -secretase, BACE1 or Memapsin-2) inhibitory activity and may therefore be used in the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and further deposits, especially alzheimer's disease.

Background

Alzheimer's Disease (AD) is a neurodegenerative disease of the central nervous system and leads to the development of progressive dementia in the middle-aged and older population. The clinical symptoms are impairment of memory, cognition, temporal and local orientation, judgment and reasoning, and severe emotional disturbances. There is currently no effective treatment that can prevent the disease or its progression or stably reverse its clinical symptoms. AD is a major health problem in all societies with a high life expectancy and also becomes a significant economic burden for these societal health systems.

The BACE1 enzyme is responsible for one of the proteolytic cleavages of the APP protein that promotes the production of Alzheimer's disease-associated A β -peptide. Preventing or preventing the production of a β -peptide by inhibiting the BACE1 enzyme is a promising therapeutic concept.

Active site-directed BACE1 inhibitors are described in, for example, WO2006/002907, and exosite-directed (catalytic domain) BACE1 inhibitors are described in, for example, Kornacker et al, Biochemistry 2005, 44, 11567-73.

Bodor et al describe modified peptides suitable for penetration of the blood brain barrier (Bodor et al Science Vol.257, 1992).

Detailed Description

Objects of the present invention are dual site BACE1 inhibitors that bind both the enzymatic active site and the catalytic domain of BACE enzymes, the preparation of the above compounds, medicaments containing them and their manufacture, as well as the use of the above compounds in the therapeutic and/or prophylactic treatment of diseases and disorders associated with the inhibition of BACE1, such as alzheimer's disease. In addition, the compounds of the invention inhibit the formation or formation and deposition of β -amyloid plaques in, on or around neural tissue (e.g., the brain) by inhibiting the production of a β from APP or an APP fragment.

The following definitions of general terms used in the present specification apply irrespective of whether the terms in question appear alone or in combination with other groups.

Table 1: amino acid abbreviations as used herein

The term "Sta" stands for statine, (3S, 4S) -4-amino-3-hydroxy-6-methylheptanoic acid (CAS 49642-07-1).

The term "MetSta" stands for (3S, 4S) -4-amino-3-hydroxy-6-methylthiohexanoic acid (CAS n/a), (Fmoc-protected CAS: 268542-18-3).

The term "27-OH-Chol" stands for 27-hydroxycholesterol (CAS 20380-11-4). See page 18, compound 4 for structure and preparation.

The term "Chol-27-TFA-ester" stands for succinamic acid (3S,8S,9S,10R,13R,14S,17R) -17- [ (1R, 5R) -1, 5-dimethyl-6- (2, 2, 2-trifluoro-acetoxy) -hexyl ] -10, 13-dimethyl-2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-1H-cyclopenta [ a ] phenanthren-3-yl ester, which is produced as a by-product during TFA cleavage of the "27-OH-Chol" -peptide.

The term "Chol 'ester'" denotes cholesterol hemisuccinate (CAS: 1510-21-0).

The term "PEG (3) stands for 12-amino-4, 7, 10-trioxadecanoic acid (CAS: 784105-33-5).

The term "PEG (4) stands for 15-amino-4, 7,10, 13, tetraoxapentadecanoic acid (CAS: n/a), (Fmoc-protected CAS: 557756-85-1).

The term "pharmaceutically acceptable salt" refers to salts suitable for use in contact with the tissues of humans and animals. Examples of suitable salts with inorganic and organic acids are, but not limited to: acetic acid, citric acid, formic acid, fumaric acid, hydrochloric acid, lactic acid, maleic acid, malic acid, methanesulfonic acid, nitric acid, phosphoric acid, p-toluenesulfonic acid, succinic acid, sulfuric acid, tartaric acid, trifluoroacetic acid (TFA), and the like. A specific salt is the trifluoroacetate salt.

The terms "pharmaceutically acceptable carrier" and "pharmaceutically acceptable auxiliary substance" refer to carriers and auxiliary substances such as diluents or excipients that are compatible with the other ingredients of the formulation.

The term "pharmaceutical composition" includes a product comprising the specified ingredients in predetermined amounts or proportions, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts. In particular, it includes products comprising one or more active ingredients, and optionally carriers comprising inert ingredients, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from decomposition of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients.

The term "half maximal inhibitory concentration" (IC)₅₀) Refers to the concentration of a particular compound required to obtain 50% inhibition of a biological process in vitro. Can be combined with IC₅₀Value-to-number conversion to pIC₅₀Value (-log IC)₅₀) Where larger values represent exponentially increasing potential. IC (integrated circuit)₅₀The values are not absolute values but depend on the test conditions, such as the concentrations employed. Can be combined with IC₅₀Values were converted to absolute inhibition constants (Ki) using the Cheng-Prusoff equation (biochem. Pharmacol. (1973) 22: 3099). The term "inhibition constant" (Ki) refers to the absolute binding affinity of a particular inhibitor for a receptor. It is measured using a competitive binding assay and is equal to the concentration at which a particular inhibitor would occupy 50% of the receptor if no competing ligand (e.g., radioligand) were present. The Ki value can be converted numerically to a pKi value (-logKi), with larger values indicating exponentially increasing potential.

By "therapeutically effective amount" is meant an amount of a compound that, when administered to a subject for the treatment of a disease state, is sufficient to effect such treatment for the disease state. The "therapeutically effective amount" will vary depending on the compound, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

The terms "as defined herein" and "as described herein" when referring to a variable incorporate by reference the broad definition of the variable as well as preferred, more preferred and most preferred definitions, if any.

The terms "treating", "contacting" and "reacting" when referring to a chemical reaction mean adding or mixing two or more reagents under suitable conditions to produce the indicated and/or desired product. It is to be understood that the reaction that produces the indicated and/or desired product may not necessarily result directly from the combination of the two reagents that were initially charged, i.e., one or more intermediates that ultimately lead to the formation of the indicated and/or desired product may be produced in the mixture.

The invention also provides pharmaceutical compositions, methods of using the compounds, and methods of making the compounds.

All individual embodiments may be combined.

The present invention relates to dual site BACE1 inhibitors that bind both the enzymatic active site and the catalytic domain of the BACE1 enzyme.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor according to claim 1, wherein the exosite (exosite) inhibitory moiety (A ') of said BACE1 inhibitor is linked to the active site inhibitory moiety (B ') through a linker (L '), or a pharmaceutically acceptable salt thereof.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor as described above wherein L' is selected from the group consisting of:

i.-(Gly)_xwhere x is 3, 4, 5 or 6,

ii-X-Gly-Gly-wherein X is selected from the group consisting of: ala, DAla, Nle, Ser, Pro, D-Pro, Lys and DLys,

-Gly-X-Gly-, wherein X is selected from the group consisting of: ala, DAla, Nle, Ser, Pro, D-Pro, Orn, Lys and DLys,

-Gly-X-, wherein X is selected from the group consisting of: ala, DAla, Nle, Ser, Pro, D-Pro, Lys and DLys,

V.-Gly-DAla-DLys-，

PEG (3), and

vii.PEG(4)。

a certain embodiment of the invention relates to a compound as described herein, wherein a' is selected from the group consisting of:

i.Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-, and

iii.Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-

a certain embodiment of the invention relates to a compound as described herein, wherein B' is selected from the group consisting of:

Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH2,

ii.Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys(Y)-NH₂wherein Y is selected from the group consisting of: palmitoyl (Palmityl), palmitoleoyl (Palmitoleic), oleoyl (Oleic), Elaidic, Erucic, isooleoyl (Vaccenic) and myristoleoyl (Myristoleic), and

Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂。

A certain embodiment of the invention relates to a compound as described herein, selected from the group consisting of:

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (oleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (elaidoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (Isooleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (erucyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (myristoleoyl) -NH2,

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (oleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (elaidoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (Isooleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (erucyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (myristoleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu- (PEG)3-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu- (PEG)4-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Ala-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DAla-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DLys-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DPro-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-A1a-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DAla-DLys-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DAla-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DPro-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Ala-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DAla-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl))-NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DLys-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DPro-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Gly-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Gly-Gly-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Lys-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Nle-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Pro-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Lys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Nle-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Om-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Pro-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Ser-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Lys-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Nle-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Pro-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂And are and

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Ser-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Or a pharmaceutically acceptable salt thereof.

A certain embodiment of the invention relates to a compound as described herein, wherein the pharmaceutically acceptable salt is trifluoroacetate salt.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor as described herein for use as therapeutically active substance.

A certain embodiment of the present invention relates to dual site BACE1 inhibitors as described herein for use as inhibitors of BACE1 activity.

A certain embodiment of the present invention relates to dual site BACE1 inhibitors as described herein for use as therapeutically active substance for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, in particular alzheimer's disease.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor as described herein for use as therapeutically active substance for the therapeutic and/or prophylactic treatment of alzheimer's disease.

A certain embodiment of the present invention relates to a pharmaceutical composition comprising a dual site BACE1 inhibitor as described herein and a pharmaceutically acceptable carrier and/or pharmaceutically acceptable auxiliary substance.

A certain embodiment of the present invention relates to the use of a dual site BACE1 inhibitor as described herein for the manufacture of a medicament for inhibiting BACE1 activity.

A certain embodiment of the present invention relates to the use of a dual site BACE1 inhibitor as described herein for the preparation of a medicament for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, in particular alzheimer's disease.

A certain embodiment of the present invention relates to the use of a dual site BACE1 inhibitor as described herein for the manufacture of a medicament for the therapeutic and/or prophylactic treatment of alzheimer's disease.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor as described herein for use in inhibiting BACE1 activity.

A certain embodiment of the present invention relates to dual site BACE1 inhibitors as described herein for use in the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, in particular alzheimer's disease.

A certain embodiment of the present invention relates to a dual site BACE1 inhibitor as described herein for use in the therapeutic and/or prophylactic treatment of alzheimer's disease.

A certain embodiment of the present invention relates to a method for use in inhibiting BACE1 activity, in particular for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, alzheimer's disease, comprising administering to a human or animal an inhibitor of dual site BACE1 as described herein.

Furthermore, the present invention includes all optical isomers, i.e. diastereoisomers, diastereoisomeric mixtures, racemic mixtures, all their corresponding enantiomers and/or tautomers as well as solvates thereof.

Dual site BACE1 inhibitors may be prepared as described herein. The starting materials are commercially available or can be prepared according to known methods.

The corresponding pharmaceutically acceptable salts with acids may be obtained by standard methods known to those skilled in the art, e.g. by dissolving a dual site BACE1 inhibitor in a suitable solvent such as diAlkane or THF and adding the appropriate amount of the corresponding acid. The product can generally be isolated by filtration or by chromatography. Conversion of a dual site BACE1 inhibitor to a pharmaceutically acceptable salt with a base can be carried out by treating such a compound with such a base. One possible method of forming such salts is, for example, by adding to a solution of the compound in a suitable solvent (e.g., ethanol-water mixtures, tetrahydrofuran-water mixtures) a 1/n equivalent of a base salt such as M (OH)_nWhere M is the metal or ammonium cation and n is the number of hydroxide anions, and removing the solvent by evaporation or freeze drying.

In the case where their preparation is not described in the examples, dual site BACE1 inhibitors as well as all intermediate products may be prepared according to analogous methods or according to the methods set forth herein. The starting materials are commercially available, known in the art or can be prepared by or analogously to methods known in the art.

It will be appreciated that dual site BACE1 inhibitors of the invention may be derivatized at functional groups to provide derivatives capable of conversion back to the parent compound in vivo.

Pharmacological testing

Dual site BACE1 inhibitors and their pharmaceutically acceptable salts possess valuable pharmacological properties. The compounds of the present invention have been found to be associated with the inhibition of BACE1 activity. The compounds were investigated according to the tests given below.

Cellular a β -reduction assay:

human HEK293 cells stably transfected with a vector expressing cDNA of the human APP wild-type gene (APP695) were used to assess the efficacy of compounds in cellular assays. Cells were seeded in cell culture medium (Iscove, plus 10% (v/v) fetal calf serum, glutamine, ampicillin/streptomycin) in 96-well microtiter plates to about 80% confluence and compounds were added at a 10x concentration in 1/10 volumes of FCS-free, 8% DMSO (final DMSO concentration maintained at 0.8% v/v) containing medium. At 37 ℃ and 5% CO₂After incubation for 18-20 hours in a wet incubator, culture supernatants are harvested for determination of A β 40 concentration.A 96-well ELISA plate (e.g., Nunc Maxisorb) is coated with a monoclonal antibody that specifically recognizes the C-terminus of A β 40 (Brockhaus et al, NeuroReport 9, 1481-1486; 1998). after blocking the non-specific binding sites with, for example, 1% BSA and washing, culture supernatants are added at appropriate dilutions with horseradish peroxidase-conjugated A β detection antibody (e.g., antibody 4G8, Senetek, Maryland Heights, MO) and incubated for 5-7 hours.subsequently the wells of the microtiter plate are washed extensively with Tris-buffered saline containing 0.05% Tween 20 and the assay is performed with tetramethylbenzidine/H₂O₂Color development was performed in citrate buffer. With a volume 1N H₂SO₄After termination of the reaction, the reaction was measured at a wavelength of 450nm in an ELISA reader the concentration of A β in the culture supernatant was calculated from a standard curve obtained with a known amount of pure A β peptide.

Table 2: IC of selected embodiments₅₀Value of

Pharmaceutical composition

The dual site BACE1 inhibitor and pharmaceutically acceptable salts may be used as therapeutically active substances, e.g. in the form of pharmaceutical formulations. Examples of pharmaceutical preparations are enteric preparations, for example in the form of tablets, coated tablets, dragees, hard and soft gelatine capsules, solutions, emulsions or suspensions and the like. However, administration may also be effected rectally, e.g. in the form of suppositories, parenterally, e.g. in the form of injection solutions, or as intranasal delivery, e.g. as intranasal sprays.

The dual site BACE1 inhibitors and pharmaceutically acceptable salts thereof may be processed with pharmaceutically inert, inorganic or organic carriers for the preparation of pharmaceutical formulations. For example, lactose, corn starch or derivatives thereof, talc, stearic acid or its salts and the like can be used as carriers for tablets, coated tablets, dragees and hard gelatine capsules. Suitable carriers for soft gelatine capsules are, for example, vegetable oils, waxes, fats, and semi-solid and liquid polyols and the like. However, depending on the nature of the active ingredient, no carriers are often required in the case of soft gelatin capsules. Suitable carriers for the preparation of solutions and syrups are, for example, water, polyols, glycerol, vegetable oils and the like. Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats and semi-liquid or liquid polyols and the like. The dual site BACE1 inhibitors and pharmaceutically acceptable salts thereof may also be encapsulated in suitable polymers or prepared using nanotechnology.

In addition, the pharmaceutical preparations can contain pharmaceutically acceptable auxiliary substances such as preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They may also contain other therapeutically valuable substances.

Medicaments containing a dual site BACE1 inhibitor or a pharmaceutically acceptable salt thereof together with a therapeutically inert carrier are also an object of the present invention, as are methods for their preparation, which comprise bringing one or more dual site BACE1 inhibitors and/or pharmaceutically acceptable salts thereof and, if desired, one or more other therapeutically valuable substances into a galenical administration form together with one or more therapeutically inert carriers.

The dosage can vary within wide limits and must, of course, be adjusted to the individual requirements in each particular case. In the case of oral administration, the dosage for adults may vary from about 0.01 mg/day to about 1000 mg/day of a dual site BACE1 inhibitor or a corresponding amount of a pharmaceutically acceptable salt thereof. The daily dose may be administered in a single dose or in divided doses, and furthermore, when necessary, the upper limit may also be exceeded.

The following examples illustrate the invention, but are not intended to be limiting and are intended to be exemplary only. The pharmaceutical formulations conveniently contain from about 1 to 500mg, preferably 1 to 100mg, of a dual site BACE1 inhibitor. Examples of compositions according to the invention are:

example A

Tablets having the following composition were prepared in the usual manner:

table 3: possible tablet compositions

The preparation process comprises the following steps:

1. ingredients 1, 2,3 and 4 were mixed and granulated with pure water.

2. The granules were dried at 50 ℃.

3. The particles are passed through a suitable milling apparatus.

4. Add ingredient 5 and mix for three minutes; pressing on a suitable press.

Example B-1

Capsules having the following composition were prepared:

table 4: possible capsule ingredient composition

The preparation process comprises the following steps:

1. ingredients 1, 2 and 3 were mixed in a suitable mixer for 30 minutes.

2. Ingredients 4 and 5 were added and mixed for 3 minutes.

3. Filling into suitable capsules.

The dual site BACE1 inhibitor, lactose and corn starch were first mixed in a mixer and then mixed in a pulverizer. Returning the mixture to the mixer; talc was added thereto and mixed well. The mixture is filled by machine into suitable capsules, for example hard gelatin capsules.

Example B-2

Soft gelatin capsules were prepared having the following composition:

composition (I)	mg/capsule
		Bi-site BACE1 inhibitors	5
Yellow wax	8
		Hydrogenated soybean oil	8
Partially hydrogenated vegetable oils	34
		Soybean oil	110
Total of	165

Table 5: possible soft gelatin capsule ingredient compositions

Table 6: possible soft gelatin capsule compositions

Preparation process

The dual site BACE1 inhibitor was dissolved in a warm melt of the other ingredients and the mixture was filled into soft gelatin capsules of appropriate size. The filled soft gelatin capsules were processed according to the general procedure.

Example C

Suppositories of the following composition were prepared:

composition (I)	mg/suppository
		Bi-site BACE1 inhibitors	15
Suppository block	1285
		Total of	1300

Table 7: possible suppository composition

Preparation process

The suppository blocks were melted in a glass or steel vessel, mixed well and cooled to 45 ℃. Thereafter, a finely powdered dual site BACE1 inhibitor was added to it and stirred until it was completely dispersed. Pouring the mixture into suppository molds with proper size, standing and cooling; the suppositories are then removed from the moulds and individually packaged in waxed paper or metal foil.

Example D

Injection solutions of the following composition were prepared:

composition (I)	mg/injection solution
		Bi-site BACE1 inhibitors	3
Polyethylene glycol 400	150
		Acetic acid	Proper amount to pH 5.0
Water for injection solution	To 1.0ml

Table 8: possible injection solution compositions

Preparation process

A dual site BACE1 inhibitor was dissolved in a mixture of polyethylene glycol 400 and water for injection (part). The pH was adjusted to 5.0 by acetic acid. The volume was adjusted to 1.0ml by adding the balance of water. The solution was filtered, filled into vials with the appropriate excess and sterilized.

Example E

Sachets (sachets) of the following composition were prepared:

composition (I)	mg/sachet
		Bi-site BACE1 inhibitors	50
Lactose, fine powder	1015
		Microcrystalline cellulose (AVICEL PH 102)	1400
Sodium carboxymethylcellulose	14
		Polyvinylpyrrolidone K30	10
Magnesium stearate	10
		Fragrance additive	1
Total of	2500

Table 9: possible sachet composition

Preparation process

The dual site BACE1 inhibitor was mixed with lactose, microcrystalline cellulose and sodium carboxymethylcellulose and granulated with a mixture of polyvinylpyrrolidone in water. The granules were mixed with magnesium stearate and flavor additives and filled into sachets.

Experimental part

The following examples are provided to illustrate the present invention. They should not be considered as limitations on the scope of the invention, but merely as representatives thereof.

For CEM free microwave peptide synthesizer (CEM Liberty) Microwave Peptide Synthesizer) of The general steps are as follows:

0.1mMol scale:

deprotection of Fmoc:

the washed and pre-swollen resin (435mg, 0.1mMol, TentaGel S RAM (Load: 0.23mMol/g), (Rapp Polymere, Cat: S30023) was treated with a solution of piperidine 20% in Dimethylformamide (DMF) (7.0mL) under microwave conditions at 50 ℃ for 3 minutes for preliminary deprotection, the resin was washed with DMF and treated with a solution of piperidine 20% in DMF (7.0mL) under microwave conditions at 75 ℃ for 5 minutes for deprotection.

Coupling of amino acids:

to the washed and pre-swollen resin was added a 0.2M solution of the amino acid in DMF (2.5mL, 5.0eq.) followed by the addition0.5M solution in DMF (1.0mL, 5.0eq.) (CAS: 1075198-30-9, Iris Biotech, Cat: RL-1175.1000) followed by the addition of Diisopropylethylamine (DIPEA)2M solution in N-methyl-2 pyrrolidone (NMP) (0.5mL, 10.0 eq.). The reaction mixture was treated at 75 ℃ for 5 minutes under microwave conditions for coupling.

025mMol scale:

deprotection of Fmoc:

the washed and pre-swollen resin (1.09g, 0.25mMol, TentaGel S RAM (Load: 0.23mMol/g), (Rapp Polymere, Cat: S30023) was treated with a solution of piperidine 20% in DMF (10.0mL) at 50 ℃ for 3 minutes under microwave conditions for initial deprotection, the resin was washed with DMF and treated with a solution of piperidine 20% in DMF (10.0mL) at 75 ℃ for 5 minutes under microwave conditions for deprotection.

Coupling of amino acids:

to the washed and pre-swollen resin was added an amino acid, 0.2M solution in DMF (5.0mL, 4.0eq.) followed by additionA solution of 0.5M in DMF (2.0mL, 4.0eq.) (CAS: 1075198-30-9, Iris Biotech, Cat: RL-1175.1000) was added followed by a solution of DIPEA 2M in NMP (1.0mL, 8.0 eq.). The reaction mixture was treated at 75 ℃ for 5 minutes under microwave conditions for coupling.

General procedure for coupling "n-C8":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with octanoic acid (79.2. mu.l, 5.0eq.), (CAS: 124-07-2, Fluka) and(214mg, 5.0eq.), (CAS: 1075198-30-9, Iris Biotech, Cat: RL-1175.1000) and a solution of DIPEA (204. mu.L, 12.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "n-C12":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with dodecanoic acid (100mg, 5.0eq.), (CAS: 143-07-7, Aldrich) and(214mg, 5.0eq.), (CAS: 1075198-30-9, Iris Biotech, Cat: RL-1175.1000) and a solution of DIPEA (204. mu.L, 12.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "n-C14":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with myristic acid (114mg, 5.0eq.), (CAS: 544-63-8, Fluka) and(214mg, 5.0eq.), (CAS: 1075198-30-9, Iris Biotech, Cat: RL-1175.1000) and a solution of DIPEA (204. mu.L, 12.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "palmitoyl":

0.1mMol scale:

will get rid ofProtected, DMF washed and pre-swollen resin with palmitoleic acid (palmitoleic acid) (102mg, 4.0eq.), (CAS: 373-49-9) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "oleoyl":

0.1mMol scale:

the deprotected, DMF-washed and pre-swollen resin was washed with oleic acid (113mg, 4.0eq.), (CAS: 112-80-1) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "elaidic acyl group":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with elaidic acid (113mg, 4.0eq.), (CAS: 112-79-8) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "Isooleoyl":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with vaccenic acid (113mg, 4.0eq.), (CAS: 693-72-1) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "erucyl":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with erucic acid (113mg, 4.0eq.), (CAS: 112-86-7) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for coupling "myristolacyl":

0.1mMol scale:

the deprotected, DMF washed and pre-swollen resin was washed with myristoleic acid (90.5mg, 4.0eq.), (CAS: 544-64-9) and(176mg, 4.0eq.), (CAS: 1075198-30-9, IrisBiotech, Cat: RL-1175.1000) and a solution of DIPEA (136. mu.L, 8.0eq.) in 5.0mL DMF were treated on a shaker at room temperature for 1 hour.

General procedure for final cleavage:

0.1mMol scale:

the resin was washed with dichloromethane (CH)₂Cl₂) Washed and then treated with a solution of trifluoroacetic acid (TFA): TIS: water 95: 2.5(5mL) on a shaker at room temperature for 30 minutes. The resin was filtered. The crude peptide was extracted with diethyl ether (Et)₂O) (35mL) was precipitated. Mixing the suspensionCentrifuge and decant the solvent. The solid was dissolved in acetonitrile and water and lyophilized to give the crude peptide.

General procedure for purification:

the crude product was dissolved in acetonitrile and water (containing 0.1% TFA) and subsequently purified by preparative HPLC. Column YMC-Actus Pro C8, 5 μm, 75X30mm, using a gradient of water (containing 0.1% TFA) to acetonitrile 70: 30 to 2: 98 at a flow rate of 30 mL/min.

All examples described in table 1 can be prepared analogously to the general procedure described herein.

Sequence listing

<110> Haofmai Roche Ltd

<120> peptide

<130>32197

<150>14195325.7

<151>2014-11-28

<160>53

<170>PatentIn version 3.5

<210>1

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>1

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>2

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>2

Tyr Pro Tyr Phe Ile Pro Leu Ala Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>3

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>3

Tyr Pro Tyr Phe Ile Pro Leu Gly Ala Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>4

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>4

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Ala Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>5

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(8)..(8)

<223> norleucine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>5

Tyr Pro Tyr Phe Ile Pro Leu Leu Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>6

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(8)..(8)

<223> norleucine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>6

Tyr Pro Tyr Phe Ile Pro Leu Gly Leu Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>7

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(10)..(10)

<223> norleucine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>7

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Leu Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>8

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>8

Tyr Pro Tyr Phe Ile Pro Leu Ser Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>9

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>9

Tyr Pro Tyr Phe Ile Pro Leu Gly Ser Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>10

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>10

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Ser Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>11

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(8)..(8)

<223> D-alanine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>11

Tyr Pro Tyr Phe Ile Pro Leu Ala Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>12

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-alanine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>12

Tyr Pro Tyr Phe Ile Pro Leu Gly Ala Gly Glu Val Asn Xaa Val Ala

1 5 1015

Glu Phe Lys

<210>13

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(10)..(10)

<223> D-alanine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>13

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Ala Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>14

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>14

Tyr Pro Tyr Phe Ile Pro Leu Pro Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>15

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>15

Tyr Pro Tyr Phe Ile Pro Leu Gly Pro Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>16

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>16

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Pro Glu Val Asn Xaa Val Ala

1 510 15

Glu Phe Lys

<210>17

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>17

Tyr Pro Tyr Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>18

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(8)..(8)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>18

Tyr Pro Tyr Phe Ile Pro Leu Pro Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>19

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>19

Tyr Pro Tyr Phe Ile Pro Leu Gly Pro Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>20

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(10)..(10)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>20

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Pro Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>21

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-alanine

<220>

<221>VARIANT

<222>(10)..(10)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>21

Tyr Pro Tyr Phe Ile Pro Leu Gly Ala Lys Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Phe Lys

<210>22

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>22

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>23

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> Lys (palmitoyl)

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>23

Tyr Pro Tyr Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>24

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(9)..(9)

<223> Z = ornithine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>24

Tyr Pro Tyr Phe Ile Pro Leu Gly Glx Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>25

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>25

Tyr Pro Tyr Phe Ile Pro Leu Lys Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>26

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>26

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Lys Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>27

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(8)..(8)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>27

Tyr Pro Tyr Phe Ile Pro Leu Lys Gly Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>28

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>28

Tyr Pro Tyr Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>29

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(10)..(10)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>29

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Lys Glu Val Asn Xaa Val Ala

1 5 1015

Glu Pro Lys

<210>30

<211>20

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(15)..(15)

<223>X=Metstatine

<220>

<221>VARIANT

<222>(19)..(19)

<223> D-phenylalanine

<220>

<221>MISC_FEATURE

<222>(20)..(20)

<223> Lys (palmitoyl)

<400>30

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Gly Gly Glu Val Asn Xaa Val

1 5 10 15

Ala Glu Phe Lys

20

<210>31

<211>21

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(16)..(16)

<223>X=Metstatin

<220>

<221>VARIANT

<222>(20)..(20)

<223> D-phenylalanine

<220>

<221>MISC_FEATURE

<222>(21)..(21)

<223> Lys (palmitoyl)

<400>31

Tyr Pro Tyr Phe Ile Pro Leu Gly Gly Gly Gly Gly Glu Val Asn Xaa

1 5 10 15

Val Ala Glu Phe Lys

20

<210>32

<211>17

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(8)..(8)

<223>Z=(PEG)4

<220>

<221>MISC_FEATURE

<222>(12)..(12)

<223>X=Metstatin

<220>

<221>VARIANT

<222>(16)..(16)

<223> D-phenylalanine

<220>

<221>MISC_FEATURE

<222>(17)..(17)

<223> Lys (palmitoyl)

<400>32

Tyr Pro Tyr Phe Ile Pro Leu Glx Glu Val Asn Xaa Val Ala Glu Phe

1 5 10 15

Lys

<210>33

<211>17

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(8)..(8)

<223>Z=(PEG)3

<220>

<221>MISC_FEATURE

<222>(12)..(12)

<223> X = Met pepstatin

<220>

<221>VARIANT

<222>(16)..(16)

<223> D-phenylalanine

<220>

<221>MISC_FEATURE

<222>(17)..(17)

<223> Lys (palmitoyl)

<400>33

Tyr Pro Tyr Phe Ile Pro Leu Glx Glu Val Asn Xaa Val Ala Glu Phe

1 5 10 15

Lys

<210>34

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>34

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>35

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoyl)

<400>35

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>36

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoleoyl)

<400>36

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>37

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (palmitoleoyl)

<400>37

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>38

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (oleoyl)

<400>38

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>39

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (oleoyl)

<400>39

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>40

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (elaidic acyl)

<400>40

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>41

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (elaidic acyl)

<400>41

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>42

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (Isooleoyl)

<400>42

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>43

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (Isooleoyl)

<400>43

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>44

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (erucyl)

<400>44

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>45

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (erucyl)

<400>45

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>46

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (myristoleoyl)

<400>46

Tyr Pro Lys Phe Ile Pro Leu Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>47

<211>19

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<220>

<221>VARIANT

<222>(9)..(9)

<223> D-lysine

<220>

<221>MISC_FEATURE

<222>(14)..(14)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(18)..(18)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(19)..(19)

<223> Lys (myristoleoyl)

<400>47

Tyr Pro Tyr Phe Ile Pro Lys Gly Lys Gly Glu Val Asn Xaa Val Ala

1 5 10 15

Glu Pro Lys

<210>48

<211>7

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<400>48

Tyr Pro Tyr Phe Ile Pro Leu

1 5

<210>49

<211>7

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<400>49

Tyr Pro Lys Phe Ile Pro Leu

1 5

<210>50

<211>7

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>VARIANT

<222>(7)..(7)

<223> D-lysine

<400>50

Tyr Pro Tyr Phe Ile Pro Lys

1 5

<210>51

<211>9

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(4)..(4)

<223> X = pepstatin

<220>

<221>MISC_FEATURE

<222>(9)..(9)

<223> Lys (palmitoyl)

<400>51

Glu Val Asn Xaa Val Ala Glu Phe Lys

1 5

<210>52

<211>10

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(4)..(4)

<223> X = pepstatin

<220>

<221>VARIANT

<222>(8)..(8)

<223> D-proline

<220>

<221>MISC_FEATURE

<222>(9)..(9)

<223> Lys (Y), Y = palmitoyl, oleoyl, elaidoyl, erucyl,

isooleoyl or myristoleoyl

<400>52

Glu Val Asn Xaa Val Ala Glu Pro Lys Tyr

1 5 10

<210>53

<211>9

<212>PRT

<213> Artificial sequence

<220>

<223> modified peptide

<220>

<221>MISC_FEATURE

<222>(4)..(4)

<223>X=MetSta

<220>

<221>VARIANT

<222>(8)..(8)

<223> D-phenylalanine

<220>

<221>MISC_FEATURE

<222>(9)..(9)

<223> Lys (palmitoyl)

<400>53

Glu Val Asn Xaa Val Ala Glu Phe Lys

1 5

Claims (13)

1. A dual site BACE1 inhibitor that binds both the enzymatic active site and the catalytic domain of the BACE enzyme.

2. The dual site BACE1 inhibitor according to claim 1, wherein the exosite inhibitory portion (A ') of the BACE1 inhibitor is linked to the active site inhibitory portion (B ') by a linker (L '), or a pharmaceutically acceptable salt thereof.

3. The dual site BACE1 inhibitor according to any one of claims 1-2, wherein L' is selected from the group consisting of:

i.-(Gly)_xwhere x is 3, 4, 5 or 6,

ii-X-Gly-Gly-wherein X is selected from the group consisting of: ala, DAla, Nle, Ser, Pro, D-Pro, Lys and DLys,

-Gly-X-Gly-, wherein X is selected from the group consisting of: ala, DAla, Nle,

ser, Pro, D-Pro, Orn, Lys and DLys,

-Gly-X-, wherein X is selected from the group consisting of: ala, DAla, Nle,

ser, Pro, D-Pro, Lys and DLys,

v.-Gly-DAla-DLys-，

PEG (3), and

vii.PEG(4)。

4. a dual site BACE1 inhibitor according to any of claims 1-3, wherein A' is selected from the group consisting of:

i.Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-, and

iii.Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-。

5. a dual site BACE1 inhibitor according to any one of claims 1-4, wherein B' is selected from the group consisting of:

Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

ii.Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys(Y)-NH₂Wherein Y is selected from the group consisting of: palmitoyl, oleoyl, elaeoyl, erucyl, isooleoyl and myristoleoyl, and

Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂。

6. The compound according to any one of claims 1 to 5, selected from the group consisting of:

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (oleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (elaidoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (Isooleoyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (erucyl) -NH₂，

Tyr-Pro-Lys-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (myristoleoyl) -NH2,

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (oleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (elaidoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (Isooleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (erucyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-DLys-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (myristoleoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu- (PEG)3-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu- (PEG)4-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Ala-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DAla-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DLys-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-DPro-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Ala-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DAla-DLys-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DAla-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DLys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-DPro-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Ala-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DAla-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DLys-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl))-NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-DPro-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Gly-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Gly-Gly-Gly-Glu-Val-Asn-MetSta-Val-Ala-Glu-DPhe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Lys-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Nle-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Pro-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Gly-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Lys-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Nle-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Orn-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Pro-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Gly-Ser-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Lys-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-DPro-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Nle-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Pro-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂And are and

Tyr-Pro-Tyr-Phe-Ile-Pro-Leu-Ser-Gly-Gly-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe-Lys (palmitoyl) -NH₂，

Or a pharmaceutically acceptable salt thereof.

7. The compound according to any one of claims 1 to 6, wherein the pharmaceutically acceptable salt is trifluoroacetate salt.

8. A dual site BACE1 inhibitor according to any one of claims 1-7 for use as therapeutically active substance.

9. A dual site BACE1 inhibitor according to any one of claims 1-7 for use as therapeutically active substance for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, in particular alzheimer's disease.

10. A pharmaceutical composition comprising a dual site BACE1 inhibitor according to any one of claims 1-7 and a pharmaceutically acceptable carrier and/or pharmaceutically acceptable auxiliary substance.

11. Use of a dual site BACE1 inhibitor according to any one of claims 1-7 for the manufacture of a medicament for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, in particular alzheimer's disease.

12. A method for inhibiting BACE1 activity, in particular for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β -amyloid levels and/or β -amyloid oligomers and/or β -amyloid plaques and other deposits, alzheimer's disease, diabetes or type 2 diabetes, comprising administering a dual site BACE1 inhibitor according to any one of claims 1-7 to a human being or animal.

13. The invention as hereinbefore described.