Fibrillar Peptides

Abstract

The present invention relates to a fibril peptide consisting of a sequence of 8 to 20 amino acids, the fibril peptide comprising the amino acid sequence RTIFIISM or a sequence at least 75% identical thereto. The present invention further relates to a method for delivering a cargo contained in a particle to a host cell, the method comprising: (a) contacting said host cell with said particle and a fibril peptide according to any one of claims 1 to 8, thereby (b) delivering said cargo to said host cell; and to uses, kits and devices related thereto.

Description

[Technical field]

[0001] The present invention relates to a fibril peptide consisting of a sequence of 8 to 20 amino acids, the fibril peptide comprising the amino acid sequence RTIFIISM or a sequence at least 75% identical thereto. The present invention further relates to a method for delivering a cargo contained in a particle to a host cell, the method comprising (a) contacting said host cell with said particle and a fibril peptide according to any one of claims 1 to 8, thereby (b) delivering said cargo to said host cell, as well as related uses, kits and devices. [Background technology]

[0002] One of the fundamental requirements of genetic technology is the ability to introduce nucleic acids into target cells with high efficiency. Many techniques have been developed for this purpose, such as naked DNA or vector systems such as viruses, among which retroviruses are widely used due to their wide spectrum of host cells and stable integration into the genome of target cells. Nevertheless, it is often desirable to further improve the infection of viruses or viral vectors. For this purpose, for example, cationic polymers such as DEAE-dextran, recombinant proteins such as RetroNectin®, and peptide-based enhancers have been used.

[0003] Peptide transduction enhancers have the advantage that they are biodegradable, easy to characterize due to their small size, and inexpensive to produce. Protamine sulfate is a mixture of highly cationic peptides derived from the sperm of different fish species, which enhances retroviral infection in a manner similar to that of cationic polymers. Protamine sulfate is an effective alternative to polybrene, as it has low cytotoxicity (Cornetta et al., J. Virol. Methods 1989, 23 (2), 187). Not only peptides as monomers, but also nanofibril-forming peptides such as Vectofusin-1, sperm-derived enhancer of viral infection (SEVI), and enhancing factor C (EF-C) lead to amplification of transduction.

[0004] Vectofusin-1 forms α-helical nanofibrils in a pH-dependent manner. It spontaneously assembles into supramolecular structures in culture medium, leading to colocalization of virus particles. This results in the sedimentation of virus particles along the cell surface of target cells (Majdoul et al., J. Biol. Chem. 2016, 291 (5), 2161). SEVI and other peptides, such as EF-C, form β-sheet nanofibrils. SEVI is the name of fibrils derived from prostatic acid phosphatase (PAP), encompassing amino acid residues 248-286. SEVI fibrils have a diameter of 5.3 ± 0.1 μm and a length of 5-10 μm (Munch et al., Cell 2007, 131 (6), 1059). These fibrils capture human immunodeficiency virus (HIV) virions and facilitate their attachment to target cells to enhance viral infection. At physiological concentrations, SEVI fibrils amplify HIV infection in various cells, e.g., T cells and macrophages. However, they also increase gene transfer by γ-RV and LV in hematopoietic and non-hematopoietic cells. This beneficial effect is also seen with VSV-G pseudotyped viral vectors (Wurm et al., J. Gene Med. 2010, 12 (2), 137). However, the expensive production of the relatively large peptide of 39 amino acid residues and the fibril formation time of at least 2 hours are limiting factors. Furthermore, long-term storage of SEVI can lead to the formation of large aggregates with low enhancing activity. EF-C (Protransduzin®-A, EP 2 452 947 A1) and Protransduzin®-B (US 2016 / 0185820) are amphipathic 12 amino acid peptides derived from the HIV-1 glycoprotein gp120. EF-C forms beta-amyloid nanofibrils with diameters of 3.4±1.1 nm, lengths ranging from 100 to 400 nm, and twists with a half-pitch of 26±2 nm. Models of EF-C nanofibrils show that at physiological pH, the lysine side chains form a hydrophilic surface with a high density of positive charges (Yolamanova et al, Nat. Nanotechnol. 2013, 8 (2), 130). Summary of the Invention [Problem to be solved by the invention]

[0005] Nonetheless, there is a continuing need for improved methods for introducing polynucleotides into host cells. [Means for solving the problem]

[0006] The above problems are addressed by the compounds, methods, uses, kits and devices having the features of the independent claims. Advantageous embodiments, which may be realised alone or in any combination, are set out in the dependent claims.

[0007] Therefore, the present invention relates to a fibril peptide consisting of a sequence of 8 to 20 amino acids, which comprises the amino acid sequence RTIFIISM or a sequence at least 75% identical thereto.

[0008] In general, terms used herein shall be given their usual and customary meanings to those skilled in the art, and shall not be limited to special or customized meanings unless otherwise indicated. When used below, the terms "have", "comprise" or "include" or any grammatical variants thereof shall be used non-exclusively. That is, these terms may refer to both the situation where no further features are present in the entity described in this context, other than the features introduced by these terms, and the situation where one or more further features are present. As an example, the expressions "A has B", "A contains B" and "A includes B" may refer to both the situation where no other elements are present in A, other than B (i.e., the situation where A is solely and exclusively composed of B), and the situation where one or more further elements are present in entity A, other than B (such as element C, elements C and D, or further elements). Also, as will be appreciated by those skilled in the art, the expressions "comprising a" and "comprising an" preferably mean "comprising one or more", i.e., are equivalent to "comprising at least one".

[0009] Furthermore, when used below, the terms "preferably", "more preferably", "most preferably", "particularly", "more particularly", "particularly", "more particularly" or similar terms are used with optional features without further limiting the possibilities. That is, the features introduced by these terms are optional features and are not intended to limit the scope of the claims in any way. The invention can be implemented by using alternative features, as the skilled person will recognize. Similarly, features introduced by "in one embodiment" or similar expressions are intended to be optional features, without any limitations on further embodiments of the invention, without any limitations on the scope of the invention, and without any limitations on the possibility of combining the features introduced in such a way with other optional or non-optional features of the invention.

[0010] As used herein, the term "standard conditions" refers to IUPAC standard ambient temperature and pressure (SATP) conditions, unless otherwise specified, i.e., preferably at a temperature of 25° C. and an absolute pressure of 100 kPa; also preferably, the standard conditions include a pH of 7. Furthermore, unless otherwise indicated, the term "about" refers to the indicated value with the technical precision generally accepted in the relevant field, preferably to the indicated value ±20%, more preferably ±10%, most preferably ±5%. Furthermore, the term "essentially" indicates that there are no deviations having an effect on the indicated result or use, i.e., possible deviations do not cause deviations of more than ±20%, more preferably ±10%, most preferably ±5% of the indicated result. In other words, "consisting essentially of" means including the specified components, but excluding other components, except for materials present as impurities, unavoidable materials present as a result of the process used to provide the components, and components added for purposes other than achieving the technical effect of the present invention. For example, a composition defined using the phrase "consisting essentially of" includes any known acceptable additives, excipients, diluents, carriers, etc. Preferably, a composition consisting essentially of a set of components will contain less than 5% by weight, more preferably less than 3% by weight, even more preferably less than 1% by weight, and most preferably less than 0.1% by weight of an unspecified component.

[0011] The degree of identity (e.g., expressed as "% identity") between two biological sequences, preferably DNA, RNA, or amino acid sequences, can be determined by algorithms well known in the art. Preferably, the degree of identity is determined by comparing two optimally aligned sequences over a comparison window, where the segments of the sequences in the comparison window may contain additions or deletions (e.g., gaps or overhangs) compared to the sequences compared for optimal alignment. The percentage is calculated by determining the number of positions at which identical residues exist in both sequences, preferably over the entire length of the polynucleotide or polypeptide, to obtain the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal alignment of sequences for comparison can be performed by the local homology algorithm of Smith and Waterman (1981), by the homology alignment algorithm of Needleman and Wunsch (1970), by the similarity search method of Pearson and Lipman (1988), by computer implementations of these algorithms (GAP, BESTFIT, BLAST, PASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, WI), or by visual inspection. Given that two sequences are specified for comparison, GAP and BESTFIT are preferably used to determine their optimal alignment, i.e., degree of identity. Preferably, default values ​​of 5.00 for gap weight and 0.30 for gap weight length are used. In the context of the biological sequences referred to herein, the term "essentially identical" denotes a percent identity value of at least 80%, preferably at least 90%, more preferably at least 98%, and most preferably at least 99%.As will be understood, the term "essentially identical" includes 100% identity. The above applies mutatis mutandis to the term "essentially complementary."

[0012] The term "fragment" of a biological macromolecule, preferably a polynucleotide or polypeptide, is used herein in a broad sense to refer to any subpart, preferably a subdomain, of the respective biological macromolecule, including the indicated sequence, structure and / or function. Thus, the term includes not only subparts generated by actual fragmentation of the biological macromolecule, but also subparts derived from the respective biological macromolecule in an abstract manner, for example in silico. Thus, as used herein, not only Fc or Fab fragments, but also, for example, single chain antibodies, bispecific antibodies and nanobodies can be referred to as fragments of immunoglobulins.

[0013] Unless otherwise indicated herein, the compounds specified, in particular the (poly)peptides and polynucleotides, may be included in a larger structure, for example covalently or non-covalently linked to each other, to carrier molecules, to retardants and other excipients. In particular, the (poly)peptides as specified may be included in a fusion polypeptide comprising further peptides, which may serve for example as a tag for purification and / or detection, as a linker or to extend the in vivo half-life of the compound. The term "detectable tag" refers to a stretch of amino acids that is added or introduced into the fusion polypeptide; preferably, the tag is added to the C-terminus or the N-terminus of the fusion polypeptide of the invention. Said stretch of amino acids preferably allows the detection of the fusion polypeptide by an antibody that specifically recognizes the tag; or preferably allows the formation of a functional conformation, such as a chelator; or preferably allows visualization, for example in the case of a fluorescent tag. Preferred detectable tags are Myc tag, FLAG tag, 6-His tag, HA tag, GST tag or fluorescent protein tag, for example GFP tag. All these tags are well known in the art. Other further peptides which are preferably included in the fusion polypeptide include further amino acids or other modifications which may serve as mediators of secretion, as mediators of blood-brain barrier crossing, as cell penetrating peptides and / or as immune stimulants. Further polypeptides or peptides to which the polypeptide may be fused are signal and / or transport sequences, or linker sequences. However, preferably, the (poly)peptide, in particular the fibril peptide, consists of the amino acids shown.

[0014] The term "polypeptide" as used herein means a molecule of several, typically at least 20, amino acids covalently linked together by peptide bonds. Molecules of fewer than 20 amino acids covalently linked by peptide bonds are generally considered to be "peptides".

[0015] The term "fibril peptide" as used herein relates to a peptide comprising (preferably consisting of) the amino acid sequence RTIFIISM (SEQ ID NO: 1) or a sequence at least 75% identical thereto, and having the property of forming fibrils in aqueous solution. Preferably, the fibril peptide comprises, preferably consists of, the amino acid sequence RTIFIISM (SEQ ID NO: 1) or a sequence at least 85% identical thereto. Preferably, the fibril peptide has a length of 8-20 amino acids, more preferably 8-16 amino acids, even more preferably 8-14 amino acids, even more preferably 8-12 amino acids, most preferably 8-10 amino acids. Preferably, the fibril peptide is a cationic peptide, i.e. having a net positive charge under standard conditions, preferably at pH 7 ... 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is a positively charged amino acid, preferably R, K or H, and X 2 is any amino acid). More preferably, the fibril peptide comprises, preferably consists of, the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1is R or K, and X is selected from A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K, and derivatives thereof. Even more preferably, the fibril peptide comprises, preferably consists of, the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X 2 is selected from the list consisting of A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K and non-negatively charged derivatives thereof, preferably X 1 is R or K, and X 2 is selected from the list consisting of A, V, L, I, M, F, T and non-negatively charged derivatives thereof. Even more preferably, the fibril peptide comprises or consists of the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9)(X 1 and X 2 is as specified herein above). Even more preferably, the fibril peptide comprises or consists of the amino acid sequence RTIFIISMX 3 X 4 (SEQ ID NO: 10), RAIFIISMX 3 X 4 (SEQ ID NO: 11), RTAFIISMX3 X 4 (SEQ ID NO: 12), RTIAIISMX 3 X 4 (SEQ ID NO: 13), RTIFAISMX 3 X 4 (SEQ ID NO: 14), RTIFIASMX 3 X 4 (SEQ ID NO:15), RTIFIIAMX 3 X 4 (SEQ ID NO: 16), RTIFIISAX 3 X 4 (SEQ ID NO: 17) (wherein X 3 and X 4 are independently selected from any amino acid, or X 3 and / or X 4 is not present, preferably X 3 is selected from the list consisting of Y, A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K and derivatives thereof, preferably non-negatively charged derivatives thereof, more preferably X 3 is Y and / or X 4 is selected from the list consisting of K, R, H, A, V, L, I, M, F, T, W, G, S, T, N, Q and derivatives thereof, preferably non-negatively charged derivatives thereof, more preferably X 4The fibril peptide preferably consists of the amino acid sequence RTIFIISM (SEQ ID NO: 1), RAIFIISM (SEQ ID NO: 18), RTAFIISM (SEQ ID NO: 19), RTIAIISM (SEQ ID NO: 20), RTIFAISM (SEQ ID NO: 21), RTIFIASM (SEQ ID NO: 22), RTIFIIAM (SEQ ID NO: 23), RTIFIISA (SEQ ID NO: 24), RTIFIISMY (SEQ ID NO: 25), RAIFIISMY (SEQ ID NO: 26), RTAFIISMY (SEQ ID NO: 27), RTIAIISMY (SEQ ID NO: 28), RTIFAISMY (SEQ ID NO: 29), RTIFIASMY (SEQ ID NO: 30), RTIFIIAMY (SEQ ID NO: 31), RTIFIISAY (SEQ ID NO: 32), RTIFIISMA (SEQ ID NO: 33), RTIFIISMYK (SEQ ID NO: 34), RAIFIISMYK (SEQ ID NO: 35), RTAFI ISMYK (SEQ ID NO: 36), RTIAIISMYK (SEQ ID NO: 37), RTIFAISMYK (SEQ ID NO: 38), RTIFIASMYK (SEQ ID NO: 39), RTIFIIAMYK (SEQ ID NO: 40), RTIFIISAYK (SEQ ID NO: 41), RTIFIISMAK (SEQ ID NO: 42), or RTIFIISMYA (SEQ ID NO: 43), more preferably consisting of the amino acid sequence RTIFIISM (SEQ ID NO: 1), RAIFIISM (SEQ ID NO: 18), RTAFIISM (SEQ ID NO: 19), RTIAIISM (SEQ ID NO: 20), RTIFAISM (SEQ ID NO: 21), RTIFIASM (SEQ ID NO: 22), RTIFIIAM (SEQ ID NO: 23), RTIFIISA (SEQ ID NO: 24), or RTIFIISMYK (SEQ ID NO: 34), preferably RTIFIISM (SEQ ID NO: 1) or RTIFIISMYK (SEQ ID NO: 34).

[0016] In a preferred embodiment, the fibril peptide comprises the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1is R or K, and X is selected from A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K, and derivatives thereof. Even more preferably, the fibril peptide comprises, preferably consists of, the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X 2 is selected from the list consisting of A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K and non-negatively charged derivatives thereof, preferably X 1 is R or K, and X 2 is selected from the list consisting of A, V, L, I, M, F, T and non-negatively charged derivatives thereof. Even more preferably, the fibril peptide comprises, preferably consists of, the amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 and X 2 is as specified hereinabove).

[0017] In a preferred embodiment, the fibril peptide comprises, preferably consists of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 18 to 43, 57 to 66, 68 to 72, 74, 76, 77, 79 to 82, 84 to 88, and 90 to 98. In a further preferred embodiment, the fibril peptide comprises, preferably consists of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 18 to 24, 34, 57 to 64, 66, 70, 72, 76, 80, 82, 84 to 88, and 90 to 98. In a further preferred embodiment, the fibril peptide comprises, preferably consists of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 18 to 24, 34, 57, 58, 60, 61, 62, 70, 72, 80, 84, 85, 86, 87, 88, 90, 91, 92, 93, 94, 95, 96, 97, and 98. In a further preferred embodiment, the fibril peptide comprises, preferably consists of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 18-34, 57, 58, 60, 62, 80, 85, 86, 87, 88, 90, 91, 92, 93, 94, 95, 96, and 97. In a further preferred embodiment, the fibril peptide comprises, preferably consists of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 34, 57, 60, 62, 85, 86, 90, 91, 92, 93, 96, and 97. In a further preferred embodiment, the fibril peptide comprises, preferably does not consist of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 67, 73, 78, 83, and 89.

[0018] The term "derivative" of an amino acid is used broadly herein and includes any organic acid, naturally occurring or non-naturally occurring, that contains an amino group and can be covalently attached to a peptide via at least one peptide bond. Preferably, the amino acid derivative is an α-amino acid, more preferably an L-amino acid, even more preferably a proteinogenic amino acid. More preferably, the amino acid derivative of an amino acid is a straight chain amino acid, preferably homoalanine, norvaline, or norleucine; a D-amino acid, preferably D-alanine, D-leucine, or D-isoleucine; a non-α-amino acid, preferably β-alanine or γ-aminobutyric acid; a derivative that includes the exchange of an atom with an atom from the same chemical group, preferably selenomethionine; and / or a naturally occurring amino acid derivative, preferably hypusine.

[0019] Fibrillar peptides have fibril-forming activity in aqueous solution, the term "fibril" relates to any thread-like structure or filament comprising, preferably consisting of, a multiplicity of fibril peptides; preferably, the fibrils are nanofibrils, i.e. having a diameter of 2-20 nm and / or a length of 100 nm-10 μm, preferably a length of 100 nm-5 μm. Preferably, the aggregation of fibril peptides occurs spontaneously in aqueous solution, where aqueous solution is a solution comprising at most 25% (v / v), preferably at most 15% (v / v), more preferably at most 10% (v / v), even more preferably at most 5% (v / v), most preferably at most 2% (v / v) of organic solvent. Methods for determining fibril-forming activity are known to the skilled person and are illustrated in the examples herein. Preferably, the fibril-forming activity is determined by a Thioflavin T binding assay, as specified herein in the examples. However, the fibril-forming activity can also be determined by transmission electron microscopy (TEM), Congo red staining, conversion measurement, and / or Fourier transform infrared (FTIR) spectroscopy, which methods are known in the art.

[0020] Preferably, the fibril peptide further has a biological activity of enhancing viral infection of host cells. Assays for establishing said biological activity are known to those skilled in the art and are shown in the examples herein. Preferably, said biological activity is determined by determining the infection efficiency of HIV in TZM-bl cells compared to a control; wherein preferably, the viral tat protein transactivates a β-galactosidase reporter gene under the control of the HIV-1-LTR promoter in TZM-bl cells. The enhancement of viral infection is preferably to cause an increase in infection rate of at least 2-fold, more preferably at least 5-fold, and even more preferably at least 10-fold, compared to a control. The fibril peptides are preferably used at a concentration of 0.01 μg / ml to 100 μg / ml, more preferably 0.05 μg / ml to 75 μg / ml, even more preferably 0.1 μg / ml to 50 μg / ml, even more preferably 0.2 μg / ml to 30 μg / ml, even more preferably 0.5 μg / ml to 25 μg / ml, and most preferably 1 μg / ml to 20 μg / ml to enhance viral infection.

[0021] Preferably, the fibril peptide is low immunogenic. The term "immunogenic" is understood by those skilled in the art. Preferably, the term relates to the induction of a response of the adaptive immune system of a subject, more preferably to a detectable response, and most preferably to a response that causes at least one systemic symptom of an immune response in a subject. Thus, the term "low immunogenic" as used herein relates to an immune response that does not cause a response that causes at least one systemic symptom of an immune response in a subject, more preferably to an immune response that is not detectable. Thus, preferably, after intramuscular administration of 0.1 μg of fibril peptide / kg body weight to a subject, 4 weeks later, in the case of a peptide with low immunogenicity, no peptide-specific antibodies and / or peptide-specific T cells are detected.

[0022] The term "subject" as used herein relates to an animal or a plant, preferably a vertebrate, more preferably a mammal, in particular a livestock animal such as a cow, a horse, a pig, a sheep and a goat, a companion animal such as a dog or a cat, or a laboratory animal such as a rat, a mouse or a guinea pig. Most preferably, the subject is a human.

[0023] The fibril peptides may be produced by any method deemed appropriate by the skilled artisan. Preferably, the fibril peptides are produced by overexpression in suitable cells, preferably bacterial or yeast cells, more preferably bacterial cells, most preferably E. coli cells. Also preferably, the fibril peptides are produced in mammalian cells, or insect cells, preferably mammalian cells, more preferably CHO cells. More preferably, the fibril peptides are produced by chemical synthesis according to methods known in the art. Preferably, the fibril peptides are purified after synthesis, for example by chromatography, preferably reversed phase chromatography, desalting, and / or other methods conventional in the art for peptide purification.

[0024] Preferably, the fibril peptide is included in a pharmaceutical composition. The terms "medicament" and "pharmaceutical composition" are in principle known to those skilled in the art. As mentioned herein, these terms relate to any composition comprising a fibril peptide, a particle comprising a cargo or a salt or prodrug thereof as a pharma-ceutically active compound, and optionally one or more other components, such as one or more pharma-ceutically acceptable excipients (e.g., pharma-ceutically acceptable carriers, etc.). The term "pharma-ceutically acceptable excipients" preferably includes any and all substances suitable for preparing a pharmaceutical composition to be administered to an organism that is not harmful. Thus, the term includes any solvent, diluent, or other vehicle, dispersing or suspending aid, surface active agent, tonicity agent, thickening or emulsifying agent, preservative, solid binder, lubricant, etc., suitable for the particular administration form desired. Examples of substances that can function as pharmaceutically acceptable excipients include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; sterile water; Ringer's solution; buffered saline; glucose solution; maltodextrin solution; ethyl alcohol; and phosphate buffer. The pharmaceutically active compound is present in the medicament in liquid or dry form, for example in lyophilized form. The pharma- ceutical active compounds are the active ingredients of medicines or drugs, preferably administered in conventional dosage forms prepared by combining the drug with standard pharmaceutical excipients according to conventional procedures, which may involve, as appropriate, mixing, granulating, compressing, and / or dissolving the ingredients to the desired preparation.It will be understood that the form and characteristics of an acceptable pharmaceutical carrier or diluent will be determined by the amount of active ingredient with which it will be combined, the route of administration, and other well-known variables. The excipient(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. In addition, the pharmaceutical composition or formulation may also include other carriers, adjuvants, preservatives, stabilizers, surfactants, buffers, and / or salts for adjusting the osmotic pressure, emulsifiers, sweeteners, colorants, flavors, and the like. Preferably, the amount of the compound in the pharmaceutical composition is the amount that is normally employed for other indications of the particular compound. The amount may vary within a wide range depending on known factors, for example, as specified herein below. However, the optimal amount can be easily and routinely determined by one skilled in the art, particularly based on existing dosage recommendations for the compound.

[0025] The route of administration of the pharmaceutical composition of the present invention depends on the particular compound used or contained in the pharmaceutical composition and follows known administration methods, for example, by inhalation, injection or infusion intravenously, parenterally, topically, rectally, nasally, buccal, vaginally, or via an implanted reservoir, or by sustained release systems. The term parenteral as used herein includes intravenous, intraperitoneal, intracerebral, intrathecal, intracranial, intramuscular, intraarticular, intrasynovial, intrasternal, intraocular, intraarterial, subcutaneous, and intradermal or local injection or infusion techniques. Preferably, the compound or pharmaceutical composition is administered to achieve a concentration sufficient to achieve an effect of treating and / or preventing a disease or disorder, i.e., to achieve an effective concentration. Furthermore, the composition can be formulated into an inhalable or injectable dosage form, such as a solution, suspension, emulsion, etc., by further adding diluents, dispersants, and surfactants. Furthermore, the compositions may be suitably formulated using any suitable method known in the art or as disclosed in Remington's Pharmaceutical Sciences (latest edition), Mack Publishing Company, Easton Pa.

[0026] The pharmaceutical composition is preferably administered in a conventional dosage form, which is prepared by combining the active compound with standard pharma- ceutically acceptable excipients and / or carriers, according to conventional procedures, as specified herein above. Therapeutically effective dose means the amount of active compound used in the pharmaceutical composition that provides the effect referred to herein. The therapeutic efficacy and toxicity of the compound can be determined by standard formulation procedures in cell cultures or experimental animals, for example, by ED50 (the dose that is therapeutically effective in 50% of the population) and LD50 (the dose that is lethal to 50% of the population), or IC50 (the dose that produces 50% inhibition). The dose ratio between the therapeutic effect and the toxic effect is the therapeutic index, which can be expressed as the ratio LD50 / ED50. The administration regimen will preferably be determined by the attending physician based on clinical factors, preferably according to any one of the methods described above. As is well known in the medical field, the dosage for any one patient may depend on a number of factors, including the patient's size, body surface area, age, the particular compound administered, sex, time and route of administration, general health, and other drugs administered at the same time. Progress can be monitored by periodic evaluation. A typical dose may be, for example, in the range of 1 μg to 1000 mg; however, doses below or above this exemplary range are envisioned, especially taking into account the factors mentioned above. In general, the regimen for regular administration of the pharmaceutical composition should be in the range of 1 μg to 100 mg units per day. If the regimen is a continuous infusion, it should also be in the range of 1 μg to 1 mg units per kg body weight per minute, respectively. Preferably, the pharmaceutical composition is administered once to the subject, i.e., it is preferably used as a one-time treatment. Depending on the subject and the mode of administration, the amount of substance administered may vary widely and provide from about 0.01 mg / kg body weight to about 100 mg / kg body weight. The pharmaceutical compositions and formulations referred to herein are administered at least once to treat or ameliorate or prevent a disease or condition described herein, however, the pharmaceutical compositions may be administered more than once, for example, 2 to 50 times, more preferably 5 to 50 times.Administration can be, for example, as an infusion or inhalation, hourly, 2-6 times a day, daily, every other day, every third day, weekly, biweekly, or monthly, preferably continuously, hourly, 2-6 times a day, or daily. Preferably, administration is adjusted to maintain an effective concentration in the subject's body for the intended period of time. Progress can be monitored by periodic evaluation. Preferably, the dosage of the compound is in accordance with the known indications of said compound, although other dosages, especially higher doses, can be envisaged.

[0027] Advantageously, it has been found in the research underlying the present invention that the fibril peptides of the present invention form fibrils that associate with certain compounds, such as virus particles, thereby strongly enhancing the uptake of such particles into host cells. Because fibril peptides are derived from host proteins, i.e. polypeptides to which hosts, particularly humans, are tolerant, fibril peptides are essentially non-immunogenic.

[0028] The definitions given above apply mutatis mutandis below. The additional definitions and explanations further given below also apply mutatis mutandis to all embodiments described herein.

[0029] The present invention further provides a method for delivering a cargo contained in a particle to a host cell, comprising the steps of: a) contacting said host cells with said particles and fibril peptides as identified herein, thereby b) delivering said cargo to said host cell; The present invention relates to a method comprising the steps of:

[0030] The method of the present invention is preferably an in vitro method. However, the method can also be used in vivo. When the method is carried out on a laboratory animal, the laboratory animal is preferably sacrificed. Furthermore, the method may comprise a step(s) in addition to the steps explicitly mentioned above. For example, further steps may relate to, for example, providing a host cell for step a) or further use of the cells produced in step b). Furthermore, one or more of said steps may be performed or assisted by an automated device.

[0031] The term "cargo" as used herein relates in principle to any compound or composition of matter suitable for packaging into a particle as specified herein below and desired to be delivered to a host cell. Thus, the cargo is preferably or consists of a chemical compound, preferably a pharmaceutical compound. More preferably, the cargo is a biological macromolecule, preferably a polynucleotide. Preferably, said polynucleotide is a viral genome or part thereof, more preferably a genetically modified viral genome or part thereof. Thus, the cargo may comprise a transgene to be delivered to the target cell. As will be appreciated by the skilled artisan, the transgene may comprise any nucleic acid sequence deemed appropriate by the skilled artisan, such as expression constructs for polypeptides and / or RNA, silencing constructs, Cas nuclease and / or gRNA expression constructs, one or more marker genes, suicide genes, etc. Preferably, the cargo is an expression construct for a chimeric antigen receptor (CAR) and preferably, the host cell is in this case a T cell.

[0032] The term "particle" as used herein relates to any particle having a suitable size to be taken up by a cell and adapted to contain or take up at least one cargo. Preferably, the particle is an anionic particle, i.e. having a negative net surface charge under standard conditions. Preferably, the particle has a size of 1 nm to 1 μm, preferably 10 nm to 500 nm, more preferably 25 nm to 400 nm, even more preferably 50 nm to 300 nm, most preferably 100 nm to 200 nm, where preferably the size is determined as the average diameter. Preferably, the particle has an outer layer comprising or consisting of at least one polypeptide. Thus, the particle may be, for example, a non-enveloped or enveloped virus, or a virus-like particle (VLP). More preferably, the particle has an outer layer comprising a lipid bilayer; thus, preferably, the particle is a liposome, for example a membrane vesicle or an artificial liposome. Preferably, the lipid bilayer of the particle comprises at least one fusogenic polypeptide adapted to mediate fusion of the lipid bilayer with a cell membrane. Preferably, said lipid bilayer is derived from a host cell; thus, preferably, the particle is an enveloped virus or a VLP thereof, preferably a retrovirus, more preferably a lentivirus, most preferably a human immunodeficiency virus (HIV) or a VLP thereof. Also preferably, the particle is selected from the list consisting of enveloped viruses, retroviruses, retroviral particles, pseudotyped retroviral particles, lentiviruses, lentiviral particles, pseudotyped lentiviral particles, VSV-G glycoprotein pseudotyped retroviral particles and / or lentiviral particles, GaLV glycoprotein pseudotyped retroviral particles and / or lentiviral particles, RD114 glycoprotein pseudotyped retroviral particles and / or lentiviral particles, MLV glycoprotein pseudotyped retroviral particles and / or lentiviral particles, and spumaviruses.As will be appreciated by those skilled in the art, the components of the particle may not even need to reach the cell interior, i.e., the cytoplasm, nucleus, and / or another cell compartment; preferably, any components of the particle, except the cargo, may not even need to reach the cell interior. Also, the cargo may only be partially delivered, e.g., only a portion of the fusion polypeptide may be delivered; and of a given number of particles, only a portion may deliver their cargo to the cell. In particular, when the particle is a virus or VLP and / or the cargo is a polynucleotide, only 0.1% or less of the particles may deliver the cargo.

[0033] As used herein, the term "host cell" preferably relates to any cell capable of receiving a cargo via a particle as specified herein above. More preferably, the host cell is capable of receiving and preferably expressing a cargo polynucleotide. Preferably, the host cell is a eukaryotic cell, preferably a yeast cell, such as a cell of a strain of baker's yeast, or an animal cell. More preferably, the host cell is an insect cell or a mammalian cell, in particular a mouse or rat cell. Most preferably, the host cell is a human cell. Preferably, the host cell is an immune cell, preferably a T cell, a cancer cell, a primary cell, a B cell, a B / T cell hybrid, a hematopoietic progenitor cell, a peripheral blood lymphocyte, a stem cell-like cell, a macrophage, a CD34+ cell, or a cell line, preferably selected from the list consisting of human 293T, HeLa, CEM-M7, TZM-bl, K562, HFF, and NIH3T3 cells.

[0034] The present invention further relates to the fibrillar peptide of the present invention for use in the treatment of a disease.

[0035] The present invention also relates to the fibrillar peptide of the invention for use in the treatment and / or prevention of cancer, genetic diseases and / or neurodegenerative diseases.

[0036] The terms "treating" and "treatment" refer to the improvement of the disease or disorder referred to herein or symptoms associated therewith to a significant extent; as used herein, the term includes the prevention of the worsening of the disease, disorder, or symptoms associated therewith. As used herein, the treating also includes the complete recovery of the health status related to the disease or disorder referred to herein. As used herein, it should be understood that treating may not be effective in all subjects to be treated. However, the term will preferably require that a statistically significant portion of subjects suffering from the disease or disorder referred to herein can be successfully treated. Whether or not the portion is statistically significant can be determined by the skilled artisan without further ado using various well-known statistical evaluation tools, such as determining confidence intervals, determining p-values, Student's t-test, Mann-Whitney test, etc. Preferred confidence intervals are at least 90%, at least 95%, at least 97%, at least 98% or at least 99%. The p-value is preferably 0.1, 0.05, 0.01, 0.005, or 0.0001. Preferably, the treatment will be effective for at least 10%, at least 20%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the subjects in a given cohort or population.

[0037] The term "preventing" means maintaining the health of a subject with respect to a disease or disorder as referred to herein for a certain period of time. It will be understood that the period of time may depend on the amount of drug compound administered and the subject's personal factors as discussed elsewhere herein. It should also be understood that prevention may not be effective in all subjects treated with the compound of the present invention. However, the term preferably requires that a statistically significant portion of the subjects of a cohort or population is effectively prevented from suffering from a disease or disorder as referred to herein or its associated symptoms. Preferably, in this context, a cohort or population of subjects is contemplated that would normally, i.e., develop a disease or disorder as referred to herein without the use of the preventive measures according to the present invention. Whether or not a portion is statistically significant can be determined without further ado by those skilled in the art using various well-known statistical evaluation tools as discussed elsewhere herein.

[0038] The term "cancer" as used herein relates to a disease of a subject characterized by uncontrolled proliferation by a group of somatic cells ("cancer cells"). This uncontrolled proliferation may lead to tumor formation, possibly accompanied by invasion and destruction of surrounding tissues (infiltration) and possibly the spread of cancer cells to other places in the body (metastasis). Preferably, the term cancer also includes recurrence. That is, preferably, the cancer is a non-solid cancer, a solid cancer, a metastasis, and / or a recurrence thereof. Preferably, the cancer is characterized by local or systemic immunosuppression, that is, preferably, the cancer is not recognized by the immune system of the subject. Preferably, the cancer treatment comprises immunotherapy, preferably cell-based immunotherapy. The term "immunotherapy" as used herein relates to the treatment of cancer by modulation of the immune response of the subject. The modulation may be to induce, enhance or suppress said immune response, for example by administration of CAR T cells obtained with fibril peptides and specifically recognizing the cancer cells. The term "cell-based immunotherapy" refers to a cancer therapy comprising the administration of immune cells, such as T cells, preferably tumor-specific NK cells, to a subject. Also preferably, the cancer treatment further comprises at least one of surgery, chemotherapy, and radiation therapy.

[0039] The term "genetic disease" as used herein relates to a disease causally linked to one or more modifications, preferably mutations, in the genome of a subject. Thus, preferably, the genetic disease is causally linked to one or more epigenetic changes, more preferably to one or more genetic mutations. Of course, the symptoms of a genetic disease are often caused by the expression of a mutated gene and / or lack of expression of a gene that provides the normal function of the gene product in one or more specific tissues and / or cell types, and treatment may include providing a non-mutated copy of the disease-related gene to the host cell. Thus, it may be preferable to genetically modify only those cells in which a mutation contributes to the disease. Preferably, the genetic disease is Duchenne muscular dystrophy, Huntington's disease, hemophilia A / B, cystic fibrosis, myotubular myopathy, glycogen storage disease, or sickle cell anemia, the causes and symptoms of which are known to the skilled artisan from medical textbooks.

[0040] The term "neurodegenerative disease" relates to a disease caused by a progressive loss of structure and / or function of neurons in the peripheral and / or central nervous system of an individual. Preferably, the neurodegenerative disease is a neurodegenerative disease of motor neurons and / or a neurodegenerative disease of the central nervous system. Preferably, the neurodegenerative disease is Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, or spinocerebellar ataxia, preferably spinocerebellar ataxia type 1 (SCA1). Of course, many neurodegenerative diseases are genetic diseases.

[0041] The present invention also relates to the fibril peptide of the present invention for use in vaccination. Preferably, the fibril peptide is in such a case non-covalently or covalently bound to an antigen. More preferably, the fibril peptide is in such a case comprised in a fusion polypeptide comprising the fibril peptide and an antigenic peptide or polypeptide, wherein the antigenic peptide or polypeptide is preferably fused to the N-terminus and / or C-terminus of the fibril peptide.

[0042] As used herein, the term "vaccination" preferably relates to administering a compound as specified herein to generate an immune response. That is, vaccination stimulates the immune system and establishes or improves immunity against infection, cancer, or any other disease associated with the de novo production of non-self antigens and / or neo-antigens in a subject. Preferably, vaccination allows to establish or improve immunity against infection. It should be understood that the vaccine according to the present invention can comprise further components, such as excipients, adjuvants, and / or further antigens. The skilled person will understand that vaccination may not generate a significant immune response in all vaccinated subjects. It should also be understood that vaccination may not be effective to prevent infection in all vaccinated subjects. However, the term requires that a preferably statistically significant portion of the subjects of a cohort or population is effectively vaccinated. Whether the portion is statistically significant or not can be determined without further ado by the skilled person using various well-known statistical evaluation tools as specified herein above. Preferably, vaccination induces a humoral immune response in the subject, i.e., induces the production of antibodies that preferably recognize, preferably specifically recognize, the antigen contained in the vaccine. The term "specifically recognize" is understood by those skilled in the art as the property of a binding agent, e.g., an antibody, to specifically bind to a certain type of molecule, while other molecules, e.g., proteins, from the same chemical class of molecules are not recognized or are recognized to a much lesser extent. Also preferably, vaccination induces a cellular immune response in the subject, i.e., the activation and / or proliferation of immune cells, preferably T cells.

[0043] The present invention also relates to the use, preferably in vitro, of the fibrillar peptides according to the invention for delivering a cargo contained in the particle to a cell.

[0044] Furthermore, the present invention relates to a pharmaceutical composition comprising the fibril peptide according to the present invention.

[0045] The present invention further relates to a kit comprising a fibril peptide according to the invention and at least one solvent and / or diluent.

[0046] The term "kit" as used herein means a collection of the above-mentioned compounds, compositions, means or reagents, which may or may not be packaged together. The components of the kit may be included by separate vials (i.e. as a kit of separate parts) or may be provided in a single vial. Furthermore, it should be understood that the kit of the present invention is preferably to be used for carrying out the method or use mentioned herein above. It is preferably contemplated that all components are provided in a ready-to-use format for carrying out the method or use mentioned herein above. Furthermore, the kit preferably comprises instructions for carrying out said method or use. The instructions can be provided by a user manual in paper or electronic form. In addition, the manual may comprise instructions for administration and / or dosage instructions using the kit of the present invention. Preferably, the kit comprises particles as specified herein above, or means for producing particles. Also preferably, the kit further comprises a cargo.

[0047] Preferably, the kit comprises a diluent and / or an administration means. Suitable diluents are described herein above; administration means are all means suitable for administering a compound to a subject, cell or tissue. Preferred administration methods are those known to those skilled in the art for the respective compound. Administration means may comprise a delivery unit for administration of the compound or composition, and a storage unit for storing the compound or composition until administration. However, it is also intended that the means of the present invention may appear as separate devices in such embodiments, and are preferably packaged together in the kit. Preferred administration means are those that can be applied without the specific knowledge of a specialized technician. In a preferred embodiment, the administration means is a syringe, more preferably a syringe with a needle, containing the compound or composition of the present invention. In another preferred embodiment, the administration means is an intravenous infusion (IV) device containing the compound or composition. In yet another preferred embodiment, the administration means is an inhaler containing the fibril peptide of the present invention, more preferably, the fibril compound is formulated for administration as an aerosol.

[0048] The present invention also relates to a device comprising the fibril peptide according to the present invention.

[0049] The term "device" as used herein relates to a system of means, including at least means operatively linked to each other to allow administration of the fibril peptide of the invention, in particular a medicament. Thus, the device is preferably adapted for administration of a compound or composition. Preferred means for administering a compound or composition vary depending on the envisaged mode of administration and are well known in the art. How the means are operatively linked varies depending on the type of means included in the device. Preferably, the means are composed of a single device. The device may include a delivery unit for administering the fibril peptide and a storage unit for storing the fibril peptide until administration. However, it is also conceivable that the means of the invention may appear as separate devices in such an embodiment, preferably packaged together as a kit. The skilled person will be able to understand how to link the means without further explanation. Preferred devices are those that can be applied without the special knowledge of a specialist. Preferably, the device is a syringe, more preferably a syringe with a needle, containing the compound or composition of the invention. Also preferably, the device is an intravenous infusion (IV) instrument containing the compound or composition. More preferably, the device is an endoscopic device comprising a compound or composition for lavaging the lung or a part thereof or a body cavity. Preferably, the device is a device for in vitro administration of fibrillar peptides to host cells, more preferably a pipetting device or a transfection device, in particular a high throughput cell processing device.

[0050] In view of the above, the following embodiments are specifically contemplated:

[0051] Embodiment 1: A fibril peptide consisting of a sequence of 8 to 20 amino acids, comprising the amino acid sequence RTIFIISM or a sequence at least 75% identical thereto.

[0052] Embodiment 2: A fibril peptide according to embodiment 1, comprising the amino acid sequence RTIFIISM (SEQ ID NO:1) or a sequence at least 85% identical thereto.

[0053] Embodiment 3: A fibril peptide according to embodiment 1 or 2, having a length of 8 to 12 amino acids, preferably 8 to 10 amino acids.

[0054] Embodiment 4: Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (sequence number 6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is a positively charged amino acid, preferably R, K or H, and X 2 The fibril peptide of any one of embodiments 1 to 3, comprising:

[0055] Embodiment 5: Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (sequence number 6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X is selected from the list consisting of A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K, and derivatives thereof.

[0056] Embodiment 6: Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (sequence number 6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X 2 is selected from the list consisting of A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K and non-negatively charged derivatives thereof.

[0057] Embodiment 7: Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X 2 The fibril peptide of any one of the preceding embodiments, comprising: A, V, L, I, M, F, T, and non-negatively charged derivatives thereof.

[0058] Embodiment 8: Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 8. The fibril peptide of any one of embodiments 1 to 7, comprising: (SEQ ID NO: 9).

[0059] Embodiment 9: Amino acid sequence RTIFIISMX 3 X 4 (SEQ ID NO: 10), RAIFIISMX 3 X 4 (SEQ ID NO: 11), RTAFIISMX 3 X 4 (SEQ ID NO: 12), RTIAIISMX 3 X 4 (SEQ ID NO: 13), RTIFAISMX 3 X 4 (SEQ ID NO: 14), RTIFIASMX 3 X 4 (SEQ ID NO:15), RTIFIIAMX 3 X 4 (SEQ ID NO: 16), RTIFIISAX 3 X 4 (SEQ ID NO: 17) (wherein X 3 and X 4 are independently selected from any amino acid, or X 3 and / or X 4 The fibril peptide according to any one of the preceding embodiments, wherein:

[0060] Embodiment 10: X 3 is selected from the list consisting of Y, A, V, L, I, M, F, T, W, G, S, T, N, Q, R, H, K and derivatives thereof, preferably non-negatively charged derivatives thereof, more preferably X 3 A fibrillar peptide according to any one of the preceding embodiments, wherein

[0061] Embodiment 11: X 4is selected from the list consisting of K, R, H, A, V, L, I, M, F, T, W, G, S, T, N, Q and derivatives thereof, preferably non-negatively charged derivatives thereof, more preferably X 4 A fibril peptide according to any one of the preceding embodiments, wherein

[0062] Embodiment 12: A fibrillar peptide according to any one of embodiments 6 to 11, wherein the derivative is a straight chain amino acid, preferably homoalanine, norvaline or norleucine; a D-amino acid, preferably D-alanine, D-leucine or D-isoleucine; a non-alpha amino acid, preferably beta-alanine or gamma-aminobutyric acid; a derivative comprising an exchange of an atom with an atom of the same chemical group, preferably selenomethionine; and / or a naturally occurring derivative of an amino acid, preferably hypusine.

[0063] Embodiment 13: Amino acid sequences RTIFIISM (SEQ ID NO: 1), RAIFIISM (SEQ ID NO: 18), RTAFIISM (SEQ ID NO: 19), RTIAIISM (SEQ ID NO: 20), RTIFAISM (SEQ ID NO: 21), RTIFIASM (SEQ ID NO: 22), RTIFIIAM (SEQ ID NO: 23), RTIFIISA (SEQ ID NO: 24), RTIFIISMY (SEQ ID NO: 25), RAIFIISMY (SEQ ID NO: 26), RTAFIISMY (SEQ ID NO: 27), RTIAIISMY (SEQ ID NO: 28), RTIFAISMY (SEQ ID NO: 29), RTIFIASMY (SEQ ID NO: 30), RTIFIIAMY (SEQ ID NO: 31), RTIFIISA (SEQ ID NO: 32), RTIFIISA (SEQ ID NO: 33), RTIFIISA (SEQ ID NO: 34), RTIFIISA (SEQ ID NO: 35), RTIFIISA (SEQ ID NO: 36), RTIFIISA (SEQ ID NO: 37), RTIAIISMY (SEQ ID NO: 38), RTIFAISMY (SEQ ID NO: 39), RTIFIASMY (SEQ ID NO: 40), RTIFIIAMY (SEQ ID NO: 41), RTIFIISA (SEQ ID NO: 42), RTIFIISA (SEQ ID NO: 43), RTIFIISA (SEQ ID NO: 44), RTIFIISA (SEQ ID NO: 45), RTIFIISA (SEQ ID NO: 46), RTIFIISA (SEQ ID NO: 47), RTIAIISMY (SEQ ID NO: 48), RTIFAISMY (SEQ ID NO: 49), RTIFIASMY (SEQ ID NO: 50), RTIFIISA (SEQ 13. The fibril peptide of any one of claims 1 to 12, consisting of: RTIFIISAY (SEQ ID NO: 32), RTIFIISMA (SEQ ID NO: 33), RTIFIISMYK (SEQ ID NO: 34), RAIFIISMYK (SEQ ID NO: 35), RTAFIISMYK (SEQ ID NO: 36), RTIAIISMYK (SEQ ID NO: 37), RTIFAISMYK (SEQ ID NO: 38), RTIFIASMYK (SEQ ID NO: 39), RTIFIIAMYK (SEQ ID NO: 40), RTIFIISAYK (SEQ ID NO: 41), RTIFIISMAK (SEQ ID NO: 42), or RTIFIISMYA (SEQ ID NO: 43).

[0064] Embodiment 14: A fibrillar peptide according to any one of embodiments 1 to 13, consisting of the amino acid sequence RTIFIISM (SEQ ID NO: 1), RAIFIISM (SEQ ID NO: 18), RTAFIISM (SEQ ID NO: 19), RTIAIISM (SEQ ID NO: 20), RTIFAISM (SEQ ID NO: 21), RTIFIASM (SEQ ID NO: 22), RTIFIIAM (SEQ ID NO: 23), RTIFIISA (SEQ ID NO: 24) or RTIFIISMYK (SEQ ID NO: 34), preferably RTIFIISM (SEQ ID NO: 1) or RTIFIISMYK (SEQ ID NO: 34).

[0065] Embodiment 15: A fibril peptide according to any one of embodiments 1 to 14, wherein all amino acids contained in the fibril peptide are L-amino acids.

[0066] Embodiment 16: A fibril peptide according to any one of embodiments 1 to 15, wherein all amino acids contained in the fibril peptide are alpha amino acids.

[0067] Embodiment 17: A fibril peptide according to any one of embodiments 1 to 16, wherein all amino acids contained in the fibril peptide are proteinogenic amino acids.

[0068] Embodiment 18: A fibril peptide according to any one of embodiments 1 to 17, having low immunogenicity, preferably being non-immunogenic to a mammal, preferably a human.

[0069] Embodiment 19: A method for delivering a cargo contained in a particle to a host cell, comprising: a) contacting said host cell with said particle and the fibril peptide according to any one of embodiments 1 to 19, thereby b) delivering said cargo to said host cell; A method comprising:

[0070] Embodiment 20: The method of embodiment 19, wherein the particle is a virus particle, a virus-like particle, or a liposome comprising at least one polypeptide of a virus.

[0071] Embodiment 21: The method of embodiment 19 or 20, wherein the virus is a retrovirus, preferably a human immunodeficiency virus.

[0072] Embodiment 22: The method of any one of embodiments 19 to 21, wherein the cargo comprised in the particle is a biological macromolecule or a pharmaceutical compound.

[0073] Embodiment 23: The method according to any one of embodiments 19 to 22, wherein the cargo comprised in the particle is a polynucleotide, preferably a viral genome or part thereof, more preferably a genetically modified viral genome or part thereof.

[0074] Embodiment 24: The method according to any one of embodiments 19 to 23, wherein said host cell is a mammalian cell, preferably a human cell.

[0075] Embodiment 25: The method according to any one of embodiments 19 to 24, wherein the host cell is a T cell, a cancer cell, a primary cell, a B cell, a B / T cell hybrid, a hematopoietic progenitor cell, a peripheral blood lymphocyte, a stem cell-like cell, a macrophage, a CD34+ cell, or a cell line, preferably selected from the list consisting of human 293T cells, HeLa cells, CEM-M7, TZM-bl, K562, HFF, and NIH3T3.

[0076] Embodiment 26: The method according to any one of embodiments 19 to 25, which is an in vitro method.

[0077] Embodiment 27: A fibrillar peptide according to any one of embodiments 1 to 18 for use in the treatment of a disease.

[0078] Embodiment 28: A fibrillar peptide according to any one of embodiments 1 to 18 for use in the treatment and / or prevention of cancer, a genetic disease or a neurodegenerative disease.

[0079] Embodiment 29: Use of a fibrillar peptide according to any one of embodiments 1 to 18 for delivering a cargo contained in the particle to a cell, preferably in vitro.

[0080] Embodiment 30: A pharmaceutical composition comprising a fibril peptide according to any one of embodiments 1 to 18.

[0081] Embodiment 31: A kit comprising a fibril peptide according to any one of embodiments 1 to 18 and at least one solvent and / or diluent.

[0082] Embodiment 32: A device comprising a fibrillar peptide according to any one of embodiments 1 to 18.

[0083] All references cited herein are hereby incorporated by reference with respect to their entire disclosure content and the disclosure content specifically mentioned in this specification. [Brief description of the drawings]

[0084] [Figure 1-1] Human interleukin-18 (IL-18) derived peptides form nanofibrils. (A) Table showing the sequence, physical and chemical parameters of IL-18 derived peptides. Positively charged amino acids are in italics. (B) ThT fluorescence of tested peptides and EF-C (Protransduzin®-A) incubated for 10 min at RT. (C) Zeta potential of peptides incubated for 10 min at RT. Mean values ​​(±SD) of duplicates from two independent experiments are shown. [Figure 1-2](D-J) Transmission electron microscopy images of EF-C (D), RK-10 (E-F), RM-8 (G-H), RS-7 (I), and FK-7 (J). Peptides were incubated at RT for 10 min (D, E, G) or at 90 °C for 30 min and then cooled overnight (F, H-J). Scale bars indicate 500 nm. au, arbitrary units; AA, amorphous aggregates; F, fibrils; MW, molecular weight; O, oligomers; RT, room temperature; SD, standard deviation; ThT, thioflavin T. [Figure 2-1] Nanofibrils formed by IL-18 peptides enhance HIV-1 infection of TZM-bl cells without showing cytotoxicity. (A) R5-tropic HIV-1 was treated with peptides and the mixture was used to infect TZM-bl cells. Tat-induced β-galactosidase activity was measured after 3 days. Numbers above the bars indicate n-fold enhancement of infection relative to the control without peptide. Absolute values ​​of the controls differed slightly, but the determining factor is the n-fold enhancement. Means (± SEM) of triplicate measurements from three independent experiments are shown. [Figure 2-2] (B) TZM-bl cells were treated with various peptide concentrations. MTT assay was performed after 3 days. For all peptide concentrations, metabolic activity in % was calculated by dividing the mean by the value of cells in the absence of peptide. Mean values ​​(±SD) of triplicate determinations from three independent experiments are shown. Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparison test. All concentrations were compared to the control (cells without peptide). Concentrations without asterisks are not significantly different from the control. *p≦0.05; RLU / s, relative light units per second; SD, standard deviation; SEM, standard error of the mean. [Diagram 3]Nanofibrils formed by IL-18 peptide enhance GALV-γ-RV infection of Jurkat cells. GALV-γ-RV was treated with peptide and the mixture was used to infect Jurkat cells. GFP-positive (GFP+) cells were determined 2 days later by flow cytometry. Numbers above the bars indicate n-fold enhancement of infection relative to the mean of controls without peptide. Absolute values ​​of controls differed slightly, but the determining factor is the n-fold enhancement. Mean values ​​(±SD) of three independent experiments are shown. GFP, green fluorescent protein. SD, standard deviation. [Figure 4] FTIR spectrum of RM-8. Values ​​in the plot refer to spectral maxima in the amide I region. RM-8 was incubated at RT for 10 min. au, arbitrary units. [Figure 5-1] RM-8 analogs from alanine scanning form positively charged nanofibrils. (A) Table showing the sequence, physical and chemical parameters of RM-8 and its analogs from alanine scanning (A1-8) and QH-9. Positively charged amino acids are in italics. Residues mutated to alanine (A) are in bold. [Figure 5-2] (B) ThT fluorescence of alanine scanning derived peptides, RM-8, QH-9 and EF-C. (C) Zeta potential of peptides. Mean values ​​(±SD) of duplicates are shown. Peptides were incubated for 10 min at RT. au, arbitrary units. SD, standard deviation. [Figure 6-1] RM-8 analogs from alanine scanning enhance HIV-1 and GALV-γ-RV infection. (A) R5-tropic HIV-1 was treated with peptides and the mixture was added to TZM-bl cells. Tat-induced β-galactosidase activity was measured 3 days later. Mean values ​​(±SD) of triplicate determinations are shown. [Figure 6-2](B) GALV-γ-RV was treated with peptides and the mixture was used to infect Jurkat cells. GFP-positive (GFP+) cells were determined 2 days later by flow cytometry. Numbers above the bars indicate n-fold enhancement of infection relative to the mean of controls without peptide. Absolute values ​​of controls differed slightly, but the determining factor is the n-fold enhancement. GFP, green fluorescent protein; RLU / s, relative light units per second; SD, standard deviation. [Figure 7-1] Comparison of the enhancing activity and cytotoxicity of RM-8 with the transduction enhancer Vectofusin®-1. (A) GALV-γ-RV was treated with peptides and the mixture was used to infect Jurkat cells. GFP-positive (GFP+) cells were determined after 2 days by flow cytometry. Numbers above the bars indicate n-fold enhancement of infection relative to the mean of controls without peptide. Mean values ​​(±SD) of three independent experiments are shown. (B) MTT assay. [Figure 7-2] (C) CellTiter-Glo® luminescent cell viability assay. Both assays were performed 2 days after peptide treatment of cells. For all peptide concentrations, metabolic activity was calculated in % by dividing the mean by the value of cells in the absence of peptide. Mean values ​​(±SD) of triplicate determinations from three independent experiments are shown. Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparison test. All concentrations were compared to control (cells without peptide). GFP, green fluorescent protein; ns, not significant; *p≦0.05; SD, standard deviation. [Figure 8]RM-8 fibrils increase lentiviral transduction of HEK293T cells more efficiently than Vectofusin®-1. (A) Lentiviral vectors pseudotyped with VSV glycoproteins were treated with peptides and the mixture was used to infect HEK293T cells. Infection rates were determined after 2 days. (B) Lentiviral vectors pseudotyped with modified RD114 / TR envelope glycoproteins were treated with peptides and the mixture was used to infect HEK293T cells. Infection rates were determined after 3 days. Numbers above the bars indicate n-fold enhancement of infection relative to the mean of controls without peptide. Means (±SD) of three independent experiments are shown. SD, standard deviation. [Figure 9] RM-8 fibrils are stable for 10 days. Fibrils were generated by dissolving lyophilized peptide in DMSO to 10 mg / ml followed by addition of PBS to a final concentration of 2 mg / ml. Fibril stocks were incubated at room temperature (RT) and 37°C and stability assays using ultra-performance liquid chromatography (UPLC) with a reversed-phase column were performed in collaboration with the Core Facility Functional Peptidomics (Ulm University) after 10 min, 1 h, 24 h and 10 days. One individual experiment is shown. [Figure 10] RM-8 nanofibrils enhance GALV-γ-RV transduction of T cells. RM-8 achieves higher transduction efficiency than Vectofusin®-1 (A) and equivalent to RetroNectin (B) after 7 days without cytotoxicity. GFP expression of CD3+ cells was analyzed by flow cytometry. Viability was determined by trypan blue staining. Human CD4+ and CD8+ T cells were isolated from buffy coats of two donors. The indicated transduction enhancers were incubated with GALV-γ-RV and then the mix was used to transduce T cells. Mean values ​​(±SD) are shown. SD, standard deviation. [Figure 11-1]RM-8 derivatives form nanofibrils and enhance GALV-γ-RV transduction into Jurkat cells. (A, B) ThT fluorescence of the tested peptides and EF-C (Protransduzin®-A) incubated for 10 min at RT. [Figure 11-2] Continued from Figure 11-1. [Figure 11-3] (C, D) GALV-γ-RV was treated with peptides and the mixture was used to infect Jurkat cells. GFP-positive (GFP+) cells were determined after 3 days by flow cytometry. Numbers above the bars indicate n-fold enhancement of infection relative to the mean of no peptide controls. Mean values ​​(±SD) of two independent experiments are shown. [Figure 11-4] Continued from Figure 11-3. [Figure 11-5] (E, F) CellTiter-Glo® luminescent cell viability assay. Assays were performed 3 days after peptide treatment of cells. Metabolic activity was calculated in % for all peptide concentrations by dividing the mean by the value of cells in the absence of peptide. Mean values ​​(±SD) of triplicate determinations from one experiment are shown. SD, standard deviation. Results are shown for peptides 1-21 (SEQ ID NOs: 57-77, A, C, and E) and peptides 22-42 (SEQ ID NOs: 78-98, B, D, and F). [Figure 11-6] Continued from Figure 11-5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0085] The following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention in any way.

[0086] Materials and Methods HIV-1 was generated from pBRNL4.39-92TH14, a plasmid containing a CCR5-tropic molecular HIV-1 clone (Papkalla et al., 2002). Infectious HIV-1 stocks were prepared by transfection into 293T cells by calcium phosphate transfection, and HIV-1 infection of TZM-bl cells was determined by detecting cellular β-galactosidase activity.

[0087] Retroviral vectors (RVs) were generated by co-transfection of a GFP-expressing murine leukemia virus (MLV) vector with E848 pCsGPpA-ed and glycoproteins derived from the gibbon ape leukemia virus (GALV). GFP-positive cells were determined by flow cytometry.

[0088] Lentiviral vectors (LV) pseudotyped with the glycoprotein of vesicular stomatitis virus (VSV-G) were generated by cotransfection of pSEW-luc2 and d8.9 and glycoproteins derived from VSV into 293T cells.

[0089] LV was generated by cotransfection into 293T cells of an env-deleted pBRHIV-1 NL4_3 derivative encoding luciferase instead of nef and phCMV-RD114 / TR encoding a chimeric envelope glycoprotein derived from feline leukemia virus (RD114) with a cytoplasmic tail derived from MLV gp.

[0090] Peptides were synthesized by KE Biochem Co.,Ltd (Shanghai City, China) and the purity was certified by Core Facility Functional Peptidomics (Ulm University). Peptides were dissolved in HFIP (1,1,1,3,3,3-hexafluoro-2-propanol) and sonicated using an ultrasonic cleaner. Peptides were stored at -20°C and freshly dissolved in DMSO to 10 mg / ml for each experiment. To generate fibrils, peptides were diluted in PBS to 2 mg / ml or 1 mg / ml and incubated at room temperature (RT) for 10 min.

[0091] Lyophilized RM-8 and its derivatives (peptides 1–42) were dissolved in DMSO to 10 mg / ml and stored at 4° C. For each experiment, they were freshly dissolved in PBS to 2 mg / ml and incubated at RT for 10 min.

[0092] For the CAR T-cell-like process, human CD4+ and CD8+ T cells were isolated from the buffy coats of two donors using RosetteSep™ human CD4+ and CD8+ T cell enrichment cocktail (STEMCELL Technologies) according to the manufacturer's protocol. The isolated T cells were activated for 3 days in the presence of 10 ng / ml interleukin-2 (IL-2) using Dynabeads™ human T-activator CD3 / CD28 (Gibco™) according to the manufacturer's protocol. After 3 days, the magnetic Dynabeads were removed using a magnet and the T cells were cultured in the presence of 9 ng / ml interleukin-7 (IL-7) and 10 ng / ml interleukin-15 (IL-15). One day later, T cells were transduced with GALV-γ-RV using the transduction enhancers RM-8 (50 μg / ml), Vectofusin®-1 (10 μg / ml), or RetroNectin (20 μg / ml). The concentrations of the transduction enhancers corresponded to those on the cells and, in the case of RetroNectin, to those used to pre-coat 24-well plates. Seven days later, transduced T cells were analyzed by flow cytometry.

[0093] result Interleukin-18 derived peptides form positively charged nanofibrils. Peptides derived from human interleukin-18 were identified by an in silico screening method in which the human protein interleukin-18 (Q14116) was analyzed by the amyloid prediction tools ZipperDB, TANGO2.0 and PASTA (Figure 1A). To investigate whether these endogenous peptides form nanofibrils, the freshly prepared peptides were incubated with the amyloid-specific dye thioflavin T (ThT), which increases in fluorescence upon binding to amyloid fibrils. ThT fluorescence scans showed that freshly prepared IL-18-derived peptides by diluting the DMSO stock (10 mg / ml) 5-fold (2 mg / ml) in PBS and incubation for 10 min at room temperature (RT) resulted in a clear ThT signal (Figure 1B). Further evidence that IL-18 peptides form nanofibrils was obtained by transmission electron microscopy (TEM) analysis, which showed fibril structures similar to the positive control EF-C (Figure 1D-J). To determine whether the IL-18 peptide has the ability to form positively charged fibrils, the zeta potential was measured. Only RK-10 and RM-8 nanofibrils exhibited a positive zeta potential like EF-C (Figure 1C).

[0094] Nanofibrils formed by IL-18 peptides enhance HIV-1 infection without exerting cytotoxicity To determine the effect on HIV-1 infection, CCR5-tropic HIV-1 stocks were generated by transient transduction of proviral DNA into 293T cells and harvested after 2 days. HIV-1 stocks were titrated and a dilution factor of 125 was selected for the infection of TZM-bl cells to reach maximum infection enhancement. HIV-1 virus was added to serially diluted IL-18 peptides and the mixture was used to infect TZM-bl cells containing the reporter gene β-galactosidase under the control of the HIV-1-LTR promoter (Wei et al., 2002). Tat-induced β-galactosidase activity was measured after 3 days and showed a dose-dependent enhancement of HIV-infection only for RK-10, RM-8 and the positive control EF-C (Figure 2A). For example, treatment of the virus with 150 μg / ml RK-10, RM-8 and EF-C enhanced HIV-1 infection 36-71 fold, whereas RS-12, RS-7 and FK-7 showed weak or no effect. To analyze possible cytotoxic effects, freshly dissolved peptides were incubated with TZM-bl cells for 3 days, and metabolic activity was then assessed by measuring the conversion of yellow MTT to purple formazan crystals by metabolically active cells. MTT assays demonstrated that none of the IL-18 peptides were cytotoxic (Figure 2B).

[0095] Nanofibrils formed by IL-18 peptide fibrils enhance GALV-γ-RV infection To investigate whether RM-8 fibrils also increase the transduction efficiency of gamma-retroviral vectors (γ-RVs) currently used for the generation of CAR T cells (Scholler et al., 2012; Muul et al., 2003; Macpherson et al., 2005), we generated RVs pseudotyped with glycoproteins derived from the gibbon ape leukemia virus (GALV-γ-RVs) encoding GFP. Serially diluted peptides were incubated with GALV-γ-RVs, and Jurkat cells were infected with this fibril mixture mix. After 2 days, GFP expression was determined by flow cytometry analysis. It was observed that treatment with RM-8, RK-10, and EF-C fibrils enhanced gene delivery rates from roughly 1% in the control to up to 20-30% GFP-positive cells, whereas other peptides exhibited only weak enhancing activity (Figure 3).

[0096] Structural characterization of the shortest peptide, RM-8 RM-8, an 8-mer endogenous peptide, was the shortest IL-18 peptide that showed potentiating activity comparable to EF-C against HIV-1 and GALV-γ-RV infection. To analyze the secondary structure of RM-8 fibrils, attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy was performed according to Sieste et al., 2021. Thus, RM-8 was freshly prepared by diluting the DMSO stock (10 mg / ml) 10-fold (1 mg / ml) in PBS and incubated at RT for 10 min. The FTIR spectrum showed peaks at approximately 1632 and 1676 cm, characteristic of the presence of a high content of intermolecular β-sheet structure. 1 The fibrillation was dominated by the amide I maximum at 10 min (Figure 4) (Zandomeneghi et al., 2004). Calculation of the native intramolecular secondary element resulted in a β-sheet content of approximately 75.9 ± 3.4%. Additionally, the conversion rate (CR) of RM-8 incubated as described above was determined according to Schilling et al., 2019. CR measures the amount of peptide monomers that form fibrils. RM-8 showed a high CR of 97.0 ± 2.6%, demonstrating high fibrillation after 10 min.

[0097] Alanine-scanning derived RM-8 analogs form positively charged nanofibrils and enhance HIV-1 and GALV-γ-RV infectivity Alanine scanning was performed to examine the function of each amino acid in RM-8 with respect to its ability to form fibrils and enhance infection. Therefore, each residue in RM-8 was mutated to alanine (Figure 5A). In addition to the RM-8 analogs (A1-A8), peptide QH-9 was selected as a negative control. QH-9 is a variant of RM-8 that lacks the N-terminal positive amino acid arginine and is negatively charged at physiological pH. To examine whether the RM-8 analogs and QH-9 form nanofibrils, the reactivity of these peptides with ThT was analyzed. ThT signals were detected for the RM-8 analogs except for A4, A8, and QH-9 (Figure 5B). Next, the zeta potential was determined. As expected, A1 and QH-9 showed negative zeta potentials due to their pI<7, while the other nanofibrils showed positive zeta potentials (Figure 5C).

[0098] Next, the enhancing effect of RM-8 analogs on HIV-1 and GALV-γ-RV infection was investigated. Therefore, HIV-1 infection assays were performed as described above. Concentration-dependent enhancement of HIV-1 infection was measured for the cationic peptides from alanine scanning (Figure 6A). The anionic peptides A1 and QH-9 slightly enhanced infection at the highest concentration.

[0099] The concentration of GALV-γ-RV particles was increased 10-fold compared to previous experiments to achieve a higher yield of GFP-positive cells. EF-C increased the number of GFP-positive cells 13-fold, from 5.4% to 72% (Figure 6B). With RM-8, a 14-fold increase in infection could be observed, which resulted in approximately 76% GFP-positive cells. Fibrils formed by cationic RM-8 analogs also increased the gene transfer rate of GALV-γ-RV, similar to RM-8. No clear infection enhancement was observed with the anionic peptides A1 and QH-9.

[0100] RM-8 increases the transduction efficiency of lentiviral and retroviral vectors pseudotyped with various glycoproteins The effect of RM-8 on transduction enhancement was analyzed for transduction with LVs and RVs with different envelopes routinely used for genetic modification, including glycoproteins from Gibbon Ape Leukemia Virus (GALV), glycoproteins from Vesicular Stomatitis Virus (VSV-G), and chimeric envelope glycoproteins from Feline Leukemia Virus (RD114) with a cytoplasmic tail from MLV gp (RD114 / TR). In addition, the enhancement activity of RM-8 for these viral particles was compared with that of Vectofusin®-1 (Radek et al., 2019), a fibrillogenic transduction enhancer integrated into the T cell transduction process in Miltenyi Biotec's Clin-iMACS Prodigy® for automated generation of T cells. Thus, GFP-encoding RVs (Figure 7A) and luciferase-encoding LVs (Figure 8A, B), pseudotyped with different glycoproteins, were combined with peptides and the mixtures were infected into target cells. Flow cytometry analysis revealed comparable infection enhancement of GALV-γ-RV for all three peptides, with the number of GFP-positive cells increasing from approximately 5% to 60-69% (Figure 7A). To determine possible cytotoxic effects of the peptides, MTT assays and CellTiter-Glo® Luminescent Cell Viability Assays were performed. The CellTiter-Glo® Luminescent Cell Viability Assay can be used to measure cell viability based on quantification of adenosine triphosphate (ATP), an indicator of metabolically active cells. None of the cell viability assays revealed cytotoxic effects of the peptides on Jurkat cells (Figure 7B, C). However, at some concentrations, a significant increase in metabolic activity was observed (Figure 7C). Luciferase assays of infected HEK293T cells performed 2 days later demonstrated that RM-8 and EF-C enhanced the transduction rate of VSV-G pseudotyped LVs, whereas Vectofusin®-1 showed no enhancing activity (Figure 8A).Similar results were obtained in RD114 / TR infection assays, with RM-8 exhibiting much higher gene delivery rates than Vectofusin®-1 (Figure 8B). These data, obtained with different lentiviral and retroviral vectors carrying different glycoproteins, indicate that RM-8 fibrils have a broad enhancing activity.

[0101] RM-8 nanofibrils are stable for 10 days Since the stability of pharmaceuticals needs to be guaranteed during application (Grassi et al., 2019), we analyzed the stability of RM-8, which contains a C-terminal methionine, an amino acid sensitive to oxidation, by ultra-performance liquid chromatography (UPLC). For that purpose, RM-8 nanofibrils were incubated at room temperature (RT) or 37 °C for 10 min, 1 h, 24 h or 10 days, followed by UPLC analysis. The chromatograms showed the same specific peaks for each time point (Figure 9), indicating that RM-8 nanofibrils are stable over time and that none of the amino acids undergo side reactions.

[0102] RM-8 nanofibrils enhance human T cell transduction as efficiently as RetroNectin RM-8 nanofibrils were tested as transduction enhancers in CAR T cell-like processes. For that purpose, activated human CD4+ and CD8+ T cells from two donors were transduced with GALV-γ-RV. The transduced T cells were cultured for 7 days and the transduction efficiency was determined by flow cytometry. RM-8 enhanced the transduction rate more efficiently than Vectofusin®-1, reaching a rate comparable to that of RetroNectin, the gold standard in the field, while showing no cytotoxicity (Figure 10). In contrast to RM-8, RetroNectin is very expensive and difficult to apply due to time-consuming coating and washing procedures.

[0103] RM-8 derivatives form nanofibrils and efficiently enhance GALV-γ-RV transduction into Jurkat cells RM-8 derivatives (peptides 1 to 39, SEQ ID NOs: 57 to 95) were synthesized by the synthesis of peptides with SEQ ID NOs: 2 to 9 (X 1 TIFIISM, RX 2 IFIISM, RTX 2 FIISM, RTIX 2 IISM, RTIFX 2 ISM, RTIFIX 2 S.M., RTIFIIX 2 M,RTIFIISX 2 ) based on X 1 Replaced with K. X 2 were replaced with I, P, G, T, K or R, respectively. In addition, two longer forms of RM-8 present in human IL-18 (peptides 40, 41; SEQ ID NOs: 96 and 97) were generated. Peptide 42 (SEQ ID NO: 98) is a variant of RM-8 containing R at positions 2 and 7. All RM-8 derivatives are shown in Table 1 herein below.

[0104] To investigate whether RM-8 derivatives (peptides 1-42) form nanofibrils, the reactivity of these peptides with ThT was analyzed. For RM-8 derivatives, except for 17, 18, 25, and 26, ThT signals were detected (Fig. 11A, B). Next, the enhancing effect of RM-8 derivatives on GALV-γ-RV infection was investigated. For that purpose, serially diluted peptides were incubated with GALV-γ-RV for 10 min, and then this mix was added to Jurkat cells. After 3 days, the transduction efficiency was determined using flow cytometry. Most RM-8 derivatives increased the number of GFP+ cells from roughly 2% to more than 20%, showing similar activity to RM-8 and EF-C (Fig. 11C, D). To determine possible cytotoxic effects of peptides, a CellTiter-Glo® luminescent cell viability assay was performed. For that purpose, Jurkat cells were incubated with diluted peptides for 3 days. Viability assays revealed no cytotoxic effects of RM-8 derivatives on Jurkat cells (FIGS. 11E, F).

[0105] Literature: Cornetta et al., J. Virol. Methods 1989, 23 (2), 187 Grassi et al., Amino Acids, 2019, 51, 1409-1431 Macpherson et al., J. Gene Med. 2005, 7 (5), 552-564. Majdoul et al., J. Biol. Chem. 2016, 291 (5), 2161 Munch et al., Cell 2007, 131 (6), 1059 Muul et al., Blood 2003, 101 (7), 2563-2569 Papkalla et al., J. Virol. 2002, 76 (16), 8455-8459 Radek et al., Hum. Gene Ther. 2019, 30 (12), 1477-1493. Schilling et al., Adv. Funct. Mater. 2019, 29, 1809112 Sieste et al., Adv. Funct. Mater. 2021, 2009382 Scholler et al., Sci. Transl. Med., 2012, 4 (132), 132ra53 Wei et al., Antimicrob. Agents Chemother. 2002, 46 (6), 1896-1905. Wurm et al., J. Gene Med. 2010, 12 (2), 137 Yolamanova et al., Nat. Nanotechnol. 2013, 8 (2), 130 Zandomeneghi et al., Protein Sci., 2009, 13, 3314-3321

[0106]

Table 1

Claims

1. A fibril peptide consisting of a sequence of 8 to 20 amino acids, comprising the amino acid sequence RTIFIISM (SEQ ID NO: 1) or a sequence at least 75% identical thereto.

2. Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is a positively charged amino acid, and X 2 The fibril peptide of claim 1, comprising:

3. The fibril peptide of claim 2, wherein X 1 is R, K or H.

4. Amino acid sequence X 1 TIFIISM (SEQ ID NO: 2), RX 2 IFIISM (SEQ ID NO: 3), RTX 2 FIISM (SEQ ID NO: 4), RTIX 2 IISM (SEQ ID NO: 5), RTIFX 2 ISM (SEQ ID NO:6), RTIFIX 2 SM (SEQ ID NO: 7), RTIFIIX 2 M (SEQ ID NO: 8), or RTIFIISX 2 (SEQ ID NO: 9) (wherein X 1 is R or K, and X 2 The fibril peptide of any one of claims 1 to 3, comprising: A, V, L, I, M, F, T, and non-negatively charged derivatives thereof.

5. 5. The fibril peptide of claim 4, wherein the derivative is a straight chain amino acid; a D-amino acid; a non-alpha amino acid; a derivative comprising an exchange of an atom with an atom of the same chemical group; or a naturally occurring derivative of an amino acid.

6. The fibrillar peptide of claim 4, wherein the derivative is homoalanine, norvaline, or norleucine; D-alanine, D-leucine, or D-isoleucine; beta-alanine or gamma-aminobutyric acid; selenomethionine; or hypusine.

7. Amino acid sequence RTIFIISMX 3 X 4 , RAIFIISMX 3 X 4 , RTAFIISMX 3 X 4 , RTIAIISMX 3 X 4 , RTIFAISMX 3 X 4 , RTIFIASMX 3 X 4 ,RTIFIIAMX 3 X 4 , RTIFIISAX 3 X 4 (In the formula, X 3 and X 4 are independently selected from any amino acid, or X 3 and / or X 4 The fibril peptide according to any one of claims 1 to 6, wherein:

8. 8. The fibrillar peptide of any one of claims 1 to 7, consisting of the amino acid sequence RTIFIISM, RAIFIISM, RTAFIISM, RTIAIISM, RTIFAISM, RTIFIASM, RTIFIIAM, RTIFIISA, RTIFIISMY, RAIFIISMY, RTAFIISMY, RTIAIISMY, RTIFAISMY, RTIFIASMY, RTIFIIAMY, RTIFIISAY, RTIFIISMA, RTIFIISMYK, RAIFIISMYK, RTAFIISMYK, RTIAIISMYK, RTIFAISMYK, RTIFIASMYK, RTIFIIAMYK, RTIFIISAYK, RTIFIISMAK, or RTIFIISMYA.

9. 9. A fibril peptide according to any one of claims 1 to 8, consisting of the amino acid sequence RTIFIISM, RAIFIISM, RTAFIISM, RTIAIISM, RTIFAISM, RTIFIASM, RTIFIIAM, RTIFIISA, or RTIFIISMYK.

10. 10. A fibril peptide according to any one of claims 1 to 9, having low immunogenicity.

11. The fibril peptide described in claim 10, which is non-immunogenic to mammals.

12. The fibril peptide described in claim 10, which is non-immunogenic to humans.

13. 1. A method for delivering a cargo contained in a particle to a host cell, comprising: a) contacting said host cells with said particles and with a fibril peptide according to any one of claims 1 to 12, thereby b) delivering said cargo to said host cell; A method comprising:

14. 14. The method of claim 13, wherein the particle is a virus particle, a virus-like particle, or a liposome comprising at least one polypeptide of a virus.

15. The method of claim 13 or 14, wherein the cargo contained in the particle is a biological macromolecule or a pharmaceutical compound.

16. 13. A composition comprising a fibril peptide according to any one of claims 1 to 12 for use in the treatment of a disease.

17. A composition comprising a fibril peptide described in any one of claims 1 to 12 for use in the treatment and / or prevention of cancer, genetic diseases, and / or neurodegenerative diseases.

18. A kit comprising a fibril peptide according to any one of claims 1 to 12 and at least one solvent or diluent.

19. A device comprising a fibrillar peptide according to any one of claims 1 to 12.

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