Medical products containing aqueous formulations of peptides

Abstract

The present invention relates to a medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide consisting of nine amino acids in a linear sequence, five of which are cationic and four of which are lipophilic, and three of which are tryptophan, one of which is non-genetically encoded. Additionally, methods of making such a product are provided, in which the aqueous formulation is introduced into the container and the filled container is mechanically sealed, as well as methods of treatment using such a product.

Description

[Technical Field]

[0001] The present invention relates to medical products containing peptides used in cancer treatment. In particular, these products can be manufactured in ready-to-use dosage forms, i.e., dosage forms that can be administered to patients immediately. Thus, these products contain peptides in stable formulations that can be transported and stored before use. [Background technology]

[0002] The prevalence of cancer in human and animal populations, and the role it plays in mortality, means that new therapies to combat tumors and tumor cells are continually needed. Removing a tumor, reducing its size, destroying its supporting vasculature, or reducing the number of cancer cells circulating in the blood or lymphatic system can be beneficial in a variety of ways, such as reducing pain or discomfort, preventing metastasis, facilitating surgical intervention, or extending lifespan.

[0003] Cancer therapies designed to combat tumors or tumor-metastatic cells typically rely on cytotoxic activity. This activity may be a cytotoxic effect of the active agent itself, or an effect indirectly used by the active agent, such as upregulating the host immune response against the tumor. These therapies are more or less selective for their target (tumor or tumor cells) over normal, healthy, or at least non-cancerous cells and tissues. Therapies with insufficient selectivity are associated with severe side effects due to exposure of normal cells to the cytotoxic activity on which the active agent relies to exert its therapeutic effect.

[0004] As described in patent publications U.S. Pat. Nos. 5,629,999, 5,729,989, 5,829,979, and 5,929,989, nine-amino acid amphipathic peptides containing cationic and tryptophan residues are highly effective antitumor agents. These peptides not only have a direct lytic effect on tumor cells by disrupting or permeabilizing cell membranes, but they are also highly effective at attacking organelles, such as mitochondria and lysosomes, causing their lysis. Disruption of the organelle membranes leads to the release of agents with potent immunostimulatory functions, commonly known as DAMPs (damage-associated molecular pattern molecules), including ATP, cytochrome C, mitochondrial CpG DNA sequences, mitochondrial formyl peptides, cathepsins (from lysosomes), and HMGB1 (from the nucleus). Consequently, these lytic peptides stimulate the immune system to provide an adaptive immune response, thereby providing long-term protection against tumor growth. The immune stimulation that occurs when tumor antigens are released by a direct lytic effect on the tumor enhances antigen presentation, providing a kind of in situ "vaccination" against the tumor.

[0005] One of these peptides is LTX-315, which is currently undergoing clinical trials for use as an anticancer agent. Currently, LTX-315 is provided in salt form as a lyophilized (freeze-dried) powder product, and storage under frozen or refrigerated conditions is recommended. LTX-315 is susceptible to hydrolytic degradation, which is typically addressed by formulating the product in nonaqueous solvents or providing it as a dried solid product, such as by lyophilization. Commercial suppliers provide LTX-315 as a dried solid product, which must be stored below room temperature. For example, Aldrich Partner Products provides LTX-315 as a powder, and its recommended storage conditions are a dark, inert atmosphere, and in a freezer below -20°C. BOC Sciences provides LTX-315 as a lyophilized powder, which is recommended for storage at -20°C. Amadis Chemical also offers LTX-315 as a powder product, which is recommended to be stored in a tightly closed container in a cool, dry place.

[0006] Because LTX-315 is generally intended to be administered by intratumoral injection, the powder must be dissolved, i.e., reconstituted, for example, in saline to obtain an isotonic solution, before it can be used for administration. Thus, a formulation for administration must be prepared in a clinical setting before use. Distribution as a lyophilized powder product that must be reconstituted before use is described in Non-Patent Document 1, which states that the lyophilized LTX-315 peptide is solubilized before use.

[0007] Additionally, protocols for clinical trials involving LTX-315 conducted in 2021 and 2022 recommended that the lyophilized powder be reconstituted with saline on-site before use. Preparing an injectable drug product (i.e., an LTX-315 solution for injection) in a clinical setting poses significantly higher risks of medication errors or microbial contamination than preparing a ready-to-use injectable drug product outside of a clinical setting. Furthermore, the reconstitution process increases the time required for product preparation and administration, complicating hospital logistics. Therefore, if LTX-315 could be safely provided as a ready-to-use aqueous formulation, this would likely be preferred over the tedious and risky on-site preparation.

[0008] Thus, it was widely recognized in the art that it would not be possible to provide a "ready to use" formulation of aqueous LTX-315.

[0009] Thus, a need exists for a convenient, ready-to-use formulation of LTX-315 that can be manufactured, shipped, and stored in a form suitable for direct administration.

[0010] The peptide sequence of LTX-315 using the single-letter amino acid code is KKWWKKW-Dip-K-NH2, where Dip = diphenylalanine. LTX-315 contains several tryptophan (W) residues, which are known to pose significant obstacles to the peptide's chemical stability in aqueous solution. Tryptophan side chains can undergo extensive chemical reactions in aqueous solution caused by exposure to light, reactive oxygen species, and to some extent pH change, elevated temperature, or exposure to metal cations, which can be extractable / leachable from primary packaging materials such as rubber stoppers and plungers [Non-Patent Document 2 and Non-Patent Document 3].

[0011] Short peptides lack the ability to form stable secondary structures in solution. This poor physical stability and the known chemical degradation pathways of similar short peptides have led to the understanding in the art that relatively short peptides, such as LTX-315, will be unstable in aqueous solution over the period between their manufacture and administration to patients. A notable chemical degradation pathway involving the tryptophan side chain, which may be expected to be relevant to LTX-315, was observed in the synthetic octapeptide octreotide, a modified analog of somatostatin (Non-Patent Document 4). In octreotide, a complex reaction pathway initiated by exposure to UV light transfers the indole moiety of D-tryptophan to the adjacent lysine (K) side chain. The original D-tryptophan amino acid is then transformed into hydroxyglycine, after which the peptide chain is cleaved into two fragments. LTX-315 has one motif with the same WK order as octreotide and two additional KW motifs.

[0012] Octreotide drug products are formulated at a maximum of only 1.0 mg / mL at approximately pH 4.0 (isoelectric point 8.3). This pH may not be favorable for intratumoral injection (a pH of 5-7 is generally acceptable for intratumoral injection). Two liquid formulations of octreotide exist: a single-use formulation and a multi-use, preserved formulation containing phenol. This product may be stored (protected from light) at 20-30°C, but only for a maximum of 14 days, indicating limited overall stability of this short peptide in aqueous formulations. Furthermore, octreotide is a cyclized peptide, which generally improves both physical and chemical stability compared to linear peptides.

[0013] Another possible chemical degradation pathway for LTX-315 is the formation of intermolecular ditryptophan crosslinks or bridges through oxidation. The formation of covalent LTX-315 dimers through ditryptophan crosslinks would be considered an undesirable degradation product. The possibility of ditryptophan formation is also highlighted through comparison with indolicidin. In this 13-residue peptide, a covalent ditryptophan crosslink was characterized between tryptophan residues 6 and 9, the same distance as between tryptophan residues 5 and 8 in LTX-315. This crosslink was formed under peptide synthesis conditions in TFA, indicating that this geometry / distance in the sequence allows for the formation of intramolecular Trp-Trp crosslinks under the correct conditions [5].

[0014] Comparisons can also be made with the decapeptide degarelix, which is commercially available as a lyophilized powder and exhibits erratic stability when formulated in aqueous solutions. Peptides of 9 or 10 amino acids are generally too short to form secondary structures that would enhance stability.

[0015] Thus, it is clear that one skilled in the art would consider LTX-315 and similar peptides, with multiple tryptophan residues and only nine amino acids in length, to be susceptible to degradation, especially in aqueous formulations. For this reason, the standard preparation of LTX-315 is a lyophilized powder product, and it has been recommended that the powder be stored in a freezer or refrigerator to prevent peptide degradation. [Prior art documents] [Patent documents]

[0016] [Patent Document 1] International Publication No. 2010 / 060497 [Patent Document 2] International Publication No. 2007 / 107748 [Patent Document 3] International Publication No. 2016 / 091487 [Patent Document 4] International Publication No. 2016 / 091490 [Non-patent literature]

[0017] [Non-Patent Document 1] Koo, DJ et al., "Biophysical Characterization of LTX-315 Anticancer Peptide Interactions with Model Membrane Platforms: Effect of Membrane Surface Charge." Int. J. Mol. Sci. 2022, 23, 10558 [Non-patent document 2] Bellmaine S, Schnellbaecher A, Zimmer A. "Reactivity and degradation products of tryptophan in solution and proteins." Free Radic Biol Med. 2020;160:696-718. doi:10.1016 / j.freeradbiomed.2020.09.002 [Non-patent document 3] Schoeneich C. “Novel chemical degradation pathways of proteins mediated by tryptophan oxidation:tryptophan side chain fragmentation.” J Pharm Pharmacol. 2018;70(5):655-665. doi:10.1111 / jphp.12688 [Non-patent document 4] National Center for Biotechnology Information (2022). PubChem Compound Summary for CID 448601, Octreotide. Retrieved June 14, 2022, from https: / / pubchem.ncbi.nlm.nih.gov / compound / Octreotide [Non-Patent Document 5] Osapay K, Tran D, Ladokhin AS, White SH, Henschen AH, Selsted ME. "Formation and characterization of a single Trp-Trp cross-link in indolicidin that confers protease stability without altering antimicrobial activity." J Biol Chem. 2000;275(16):12017-12022. doi:10.1074 / jbc.275.16.12017 Summary of the Invention [Means for solving the problem]

[0018] However, the present inventors have surprisingly found that peptides, such as LTX-315 and its pharmaceutically acceptable salts, are in fact stable in aqueous solution. This means that peptide-containing products can be manufactured in ready-to-use dosage forms, i.e., dosage forms that can be administered directly to patients. These products contain the peptide in a stable form that can be transported and stored prior to use.

[0019] The present invention therefore provides a medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide comprising: a) consists of a linear sequence of 9 amino acids, b) 5 of those 9 amino acids are cationic and 4 are lipophilic, and c) three of these four lipophilic amino acids (i.e., amino acids with a lipophilic R group) are tryptophan, one is non-genetically encoded, and optionally d) the lipophilic and cationic residues are arranged such that no more than three of either type of residue are adjacent to each other, and further optionally e) The molecule contains two pairs of adjacent cationic amino acids and one or two pairs of adjacent lipophilic residues.

[0020] Such formulations are suitable for administration directly to a patient, e.g., into a tumor, e.g., by injection or perfusion / infusion, etc. This provides improved convenience for clinicians who do not need to prepare a formulation for administration to a patient, e.g., by dissolving the peptide in powder form in a suitable solvent.

[0021] peptide The cationic amino acids may be the same or different and are preferably lysine or arginine, but may also be histidine or any non-genetically encoded or modified amino acid that has a positive charge at pH 7.0. Lysine is particularly preferred.

[0022] Suitable non-genetically encoded cationic amino acids and modified cationic amino acids include analogs of lysine, arginine, and histidine, such as homolysine, ornithine, diaminobutyric acid, diaminopimelic acid, diaminopropionic acid, and homoarginine, as well as trimethylysine and trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-carbamimidoylpiperidine-4-carboxylic acid, and 4-guanidinophenylalanine.

[0023] Non-genetically encoded lipophilic amino acids have R groups with at least seven, preferably at least eight or nine, and more preferably at least ten non-hydrogen atoms. Amino acids with lipophilic R groups are referred to herein as lipophilic amino acids or lipophilic residues. Typically, lipophilic R groups have at least one, and preferably two, cyclic groups, which may be fused or connected.

[0024] The lipophilic R group may contain heteroatoms such as O, N, or S, but typically no more than one heteroatom will be present, which is preferably nitrogen. The R group will preferably have no more than two polar groups, more preferably zero or one, and most preferably none.

[0025] As disclosed herein, D amino acids are not strictly genetically encoded, but are not considered "non-genetically encoded amino acids" which should differ structurally as well as stereospecifically from the 20 genetically encoded L amino acids. Some or all of the amino acids in the molecules disclosed herein may exist in the D form.

[0026] The R group of the non-genetically encoded lipophilic amino acid preferably contains 30 or fewer non-hydrogen atoms, more preferably 25 or fewer non-hydrogen atoms.

[0027] Preferred non-genetically encoded amino acids are 2-amino-3-(biphenyl-4-yl)propanoic acid (biphenylalanine), 2-amino-3,3-diphenylpropanoic acid (diphenylalanine), 2-amino-3-(anthracen-9-yl)propanoic acid, 2-amino-3-(naphthalen-2-yl)propanoic acid, 2-amino-3-(naphthalen-1-yl)propanoic acid, 2-amino-3-[1,1':4',1"-terphenyl-4-yl]-propionic acid, 2-amino-3-(2,5,7-tri-tert-butyl-1H-indoline), ... Examples of amino acids include 2-amino-3-[1,1':3',1"-terphenyl-4-yl]-propionic acid, 2-amino-3-[1,1':2',1"-terphenyl-4-yl]-propionic acid, 2-amino-3-(4-naphthalen-2-yl-phenyl)-propionic acid, 2-amino-3-(4'-butylbiphenyl-4-yl)propanoic acid, 2-amino-3-[1,1':3',1"-terphenyl-5'-yl]-propionic acid, and 2-amino-3-(4-(2,2-diphenylethyl)phenyl)propanoic acid. Diphenylalanine and biphenylalanine are particularly preferred.

[0028] In preferred embodiments, the peptide has one of the formulas (I) to (V) listed below, where C represents a cationic amino acid as defined above, and L represents a lipophilic amino acid as defined above (i.e., tryptophan (Trp / W) or a non-genetically encoded lipophilic amino acid). The amino acids are covalently linked by a peptide bond. The free amino or carboxy termini of these molecules may be modified, with the carboxy terminus preferably being modified to remove the negative charge, and most preferably the carboxy terminus being amidated, with the amide group optionally being substituted. CCLLCCLLC(I) (SEQ ID NO: 1) LCCLLCCLC(II) (SEQ ID NO: 2) CLLCCLLCC(III) (SEQ ID NO: 3) CCLLCLLCC(IV) (SEQ ID NO: 4) CLCCLLCCL(V) (SEQ ID NO: 5)

[0029] The term "amino acid" includes beta and gamma amino acids as well as alpha amino acids, and also includes N-substituted glycines.

[0030] As discussed above, the peptide includes one non-genetically encoded lipophilic amino acid. When this residue is designated L', preferred peptides are represented by the formula: CCL'LCCLLC (I') (SEQ ID NO: 6) CCLLCCLL'C (I”)(SEQ ID NO:7) CCLL'CCLLC (I”')(SEQ ID NO:8) LCCLL'CCLC (II') (SEQ ID NO: 9)

[0031] Particularly preferred are peptides of formula (I) and (II), of which the peptide of formula (I") is particularly preferred.

[0032] The following peptides are most preferred:

[0033] [Table 1]

[0034] where Standard single-letter codes are used for genetically coded amino acids Lowercase letters indicate D amino acids Dip is diphenylalanine Bip is biphenylalanine Orn is ornithine Dap is 2,3-diaminopropionic acid Dab is 2,4-diaminobutyric acid 1-Nal is 1-naphthylalanine 2-Nal is 2-naphthylalanine Ath is 2-amino-3-(anthracen-9-yl)propanoic acid Phe(4,4'Bip) is 2-amino-3-[1,1':4',1"-terphenyl-4-yl]propionic acid

[0035] Preferred peptides are LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329. The preferred peptides are preferably in the form of a pharmaceutically acceptable salt, preferably an acetate salt.

[0036] For use in the present invention, all peptides described herein may be in the form of pharmaceutically acceptable salts.Peptides preferably have modified, particularly amidated, C-terminus.Suitable pharmaceutically acceptable salts are well known in the art, including inorganic or organic acid salts, including hydrochloride, trifluoroacetate, and acetate.Acetate is most preferred.

[0037] Particularly preferred is the peptide known as LTX-315, especially in the form of a pharmaceutically acceptable salt, preferably the acetate salt thereof.

[0038] Thus, in a preferred embodiment, the present invention provides a medical product in the form of a sealed container containing an aqueous formulation of LTX-315 having the amino acid sequence of SEQ ID NO:23 or a pharmaceutically acceptable salt thereof, preferably the acetate salt thereof.

[0039] Peptide synthesis The peptides described herein, such as LTX-315 and its pharmaceutically acceptable salts, may be synthesized in any convenient manner by methods of peptide synthesis well known in the art, preferably by carrying out the synthesis on a solid support. Synthetic methods are described, for example, in U.S. Patent Nos. 5,629,999; 5,629,999; and 5,629,999.

[0040] aqueous preparations As discussed above, the medical products of the present invention are in the form of a sealed container containing an aqueous formulation of a peptide as defined herein. In the discussion that follows, the term "peptide" refers to a peptide and its pharmaceutically acceptable salts.

[0041] The sealed container may contain only an aqueous formulation of a peptide as defined herein, hi other embodiments, the sealed container is not completely filled with an aqueous formulation of a peptide as defined herein, and the unfilled volume (headspace) contains air or an inert gas such as, for example, nitrogen or argon.

[0042] In some embodiments, the aqueous formulation contains the peptide at a concentration of 1-30 mg / ml, preferably 4-20 mg / ml, or more preferably 6-16 mg / ml, where the amounts of peptide referred to above refer to net peptide amounts (see the Examples section for an explanation of net peptide amounts).

[0043] Aqueous formulations are those in which the solvent for dissolving the peptide is water, ie, aqueous solutions of the peptide.

[0044] Because peptides are readily soluble in water, the aqueous formulation does not need to include an organic co-solvent to dissolve the peptide, and indeed the aqueous formulation may be completely free of organic components (other than the peptide), although organic components may be included as, for example, preservatives, viscosity enhancers, or stabilizers, as discussed below.

[0045] The aqueous formulation may consist solely of water and peptide, i.e., the aqueous formulation is an aqueous solution of the peptide, such as in water for injection (WFI). Alternatively, the aqueous formulation may comprise or consist of water, peptide, and a salt (e.g., sodium chloride), thereby providing a saline solution of the peptide. In some other embodiments, the aqueous formulation comprises or consists of the peptide and an aqueous buffer, i.e., the aqueous formulation is a solution of the peptide in an aqueous buffer. In some embodiments, the aqueous buffer is a mixture of several different aqueous buffers. Preferably, the aqueous formulation comprises a solution of the peptide in an aqueous buffer, and optionally one or more additional excipients as discussed below.

[0046] In some embodiments, the aqueous formulation does not contain preservatives and / or stabilizers, hi some embodiments, the aqueous formulation does not contain hydroxybenzoic acid and / or EDTA.

[0047] Optionally, the aqueous formulation may contain one or more additional pharmaceutically acceptable components, i.e., excipients, such as those used in medical products containing aqueous formulations of active ingredients, which are well known in the art. Suitable excipients are, for example, those used in parenteral formulations, such as those administered by intratumoral injection, which is the preferred method of administration in the context of the present invention.

[0048] For example, the aqueous formulation may contain one or more preservatives, especially when the sealed container contains multiple doses. The following table lists examples of preferred preservatives. The preferred concentration ranges for these agents when present in the aqueous formulation are also listed.

[0049] [Table 2]

[0050] Benzyl alcohol is a particularly preferred preservative, which is preferably present at 0.75% to 2% w / w, preferably 1% w / w. Another preferred preservative is phenol, which is preferably present at 0.15% to 0.5%.

[0051] One or more chelating agents may be included to chelate metal ions. The following table lists examples of preferred chelating agents. The preferred concentration ranges for these agents in the aqueous formulation are also listed.

[0052] [Table 3]

[0053] Preferably, the chelating agent is disodium EDTA and / or sodium EDTA. Preferably, the chelating agent, e.g., disodium or sodium EDTA, is present at about 0.1% w / w.

[0054] One or more antioxidants may be included. The following table lists examples of preferred antioxidants. Also listed are suitable concentration ranges for the antioxidants when present in the aqueous formulation.

[0055] [Table 4]

[0056] A preferred antioxidant is thiourea.

[0057] As mentioned above, in some embodiments, the aqueous formulation is a solution of the peptide in an aqueous buffer or a mixture of aqueous buffers. A buffer may be used to maintain the pH of the aqueous formulation within a desired range. Preferably, the pH of the aqueous formulation is 5.0 to 7.4, more preferably 5.0 to 7.0, and even more preferably 5.0 to 6.0 or 6.5. Accordingly, the following aqueous buffers are particularly preferred: acetic acid, acetate, aspartic acid, sodium benzoate, benzoic acid, carbonic acid, citric acid, glucono-delta-lactone, glycine, glycine HCl, histidine, histidine HCl, hydrobromic acid, phosphoric acid (e.g., PBS), sodium succinate, disodium succinate, succinic acid, sulfuric acid, tartaric acid, and / or sodium tartrate. Preferably, the aqueous buffer is an acetic acid / acetate buffer, a citrate buffer, and / or a phosphate buffer.

[0058] One or more tonicity modifiers, such as, for example, dextrose, glycerin, lactose, mannitol, potassium chloride, sodium chloride, and / or sorbitol, may be included to provide an isotonic aqueous formulation that reduces pain experienced by a patient receiving the aqueous formulation by parenteral administration. Preferably, the tonicity modifier is sodium chloride.

[0059] In some embodiments, the aqueous formulation contains one or more stabilizers that inhibit peptide aggregation. Such stabilizers include anti-stacking agents. Examples of suitable stabilizers are dimethyl sulfoxide, dimethylacetamide, ethanol, glycerol, mannitol, N-methyl-2-pyrrolidone, polyethylene glycol (PEG) 200, PEG 300, PEG 350, PEG 400, PEG 600, propylene glycol, and sorbitol. PEG stabilizers, such as PEG 200, are particularly preferred. Such stabilizers, such as PEG stabilizers, may be present at 1-8% w / w, preferably 2-7% w / w, such as 3-6% w / w.

[0060] The aqueous formulation may further comprise one or more viscosity enhancing agents, such as gum acacia, agar, cellulose derivatives such as methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose, calcium carboxymethylcellulose, sodium carboxymethylcellulose, chitosan, gelatin, hyaluronic acid, maltodextrin, pectin, poloxamer 407, PEG, polyethylene oxide, povidone (PVP), starches such as corn starch and tapioca starch, tragacanth, and / or β-cyclodextrin.

[0061] By increasing the viscosity of aqueous formulation (compared to water), it can effectively slow down the dispersion of peptide from the administration site.When the formulation is administered into tumor, increasing viscosity can prolong the time that the formulation stays in tumor and does not leak out, slow down the dispersion of peptide by blood, and increase the time that peptide directly contacts tumor cells.In this way, the efficacy of peptide can be increased and / or toxicity can be reduced.

[0062] Thus, in some embodiments, the aqueous formulation comprises one or more viscosity enhancing agents and has a viscosity of from 1.5, 2, 2.5, or 3 cP to 300 cP, such as 2.5-250 cP, such as 5-100 cP or 10-200 cP and 20-240 cP, preferably 40-180 cP, such as 45-150 cP or 30-100 cP, more preferably 1.5-10 cP, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 cP, or 10-15 cP or 15-20 cP or 20-35 cP or 35-50 cP or 50-100 cP.

[0063] In some embodiments, when the medical product is a pre-filled syringe or a pre-filled syringe cartridge and the viscosity of the aqueous formulation of the medical product is greater than 250 cP, greater than 200 cP, greater than 150, or greater than 100 cP, a specialized needle may be used.

[0064] Preferably, the one or more viscosity-enhancing agents are selected from the group consisting of chitosan or a chitosan derivative or a pharmaceutically acceptable salt thereof, such as the hydrochloride salt, hydroxyethylcellulose, hydroxypropylcellulose, HPMC, poloxamer 407, PEG, and PVP. HPMC and chitosan are particularly preferred viscosity-enhancing agents. Chitosan may be ultra-low molecular weight chitosan (typically <50 kDa), low molecular weight chitosan (typically 50 to <100 kDa), medium molecular weight chitosan (typically 100-1000 kDa), or high molecular weight chitosan (typically >1000 kDa). In some embodiments, the degree of deacetylation (DDA) of the chitosan is about 50-99%, e.g., 70-99%, e.g., 80%, preferably about 70% or greater.

[0065] Chitosan contains reactive functional groups that are susceptible to chemical reactions, and modifications of chitosan, such as acylation, carboxylation, alkylation, and quaternization, have resulted in chitosan derivatives that have properties different from chitosan, such as improved solubility or bioactivity, while retaining the unique original pharmacological properties of chitosan (see Wang et al., Int. J. Mol. Sci. 21(2), 2020, 487, doi:10.3390 / ijms21020487).

[0066] In some embodiments, the viscosity enhancer is a chitosan derivative, such as O- and N-acylated chitosan, O- and N-alkylated chitosan, or chitosan modified by the introduction of hydrophilic groups, such as carboxylic acid groups, quaternary ammonium groups, sulfonic acid groups, phosphate groups, amino groups, ether bonds composed of oxygen groups, hydroxyl groups, carboxylic acid groups, and block polyether groups.

[0067] In some embodiments, the viscosity enhancing agent is a glycated chitosan obtained by reaction of the free amino groups of chitosan with the carbonyl groups of reducing monosaccharides and / or oligosaccharides. Examples of such monosaccharides include naturally occurring D-trioses, D-tetroses, D-pentoses, D-hexoses, and D-heptoses, such as D-glucose, D-galactose, D-fructose, D-mannose, D-allose, D-altrose, D-idose, D-talose, D-fucose, D-arabinose, D-gulose, D-hammelose, D-lyxose, D-ribose, D-rhamnose, D-threose, D-xylose, D-psicose, D-sorbose, D-tagatose, D-glyceraldehyde, dihydroxyacetone, D-erythrose, D-threose, D-erythrulose, D-mannoheptulose, and D-sedoheptulose. Examples of such oligosaccharides include fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), and mannan-oligosaccharides (MOS). An example of glycated chitosan is galacto-chitosan. Glycated chitosan and its synthesis are described in, for example, U.S. Pat. No. 5,747,475 (A) and International Publication Nos. 2002 / 040055 (A2) and 2013 / 109732 (A2), the contents of which are incorporated herein by reference.

[0068] In some embodiments, the viscosity enhancing agent is a glycated chitosan having a percent glycation of its otherwise free amino groups of about 0.1% to about 90%, e.g., about 0.1% to about 30%, or for example, about 2-15%, such as about 12.5%. In some embodiments, the molecular weight of the glycated chitosan is about 50 kD to about 2000 kD, e.g., about 50 kD to about 1500 kD, such as about 250 kD or 300 kD. In some embodiments, the DDA of the glycated chitosan is about 50-99%, e.g., 70-99%, such as 80%. In some embodiments, the DDA of the glycated chitosan is about 50-99%, such as 70-99%, such as 80%, and it has a glycation rate of about 0.1% to about 90% of its otherwise free amino groups, such as about 0.1% to about 30%, or such as about 2-15%, such as about 12.5%, and its molecular weight is about 50 kD to about 2000 kD, such as about 50 kD to about 1500 kD, such as about 250 kD or about 300 kD.

[0069] The viscosity enhancing agent is preferably present at 0.1-5% w / w, such as 0.15-2.5% w / w or 0.2-2% w / w or 0.25-1.5% w / w or 0.3-1% w / w or 0.35-0.9% w / w, etc. In some embodiments, the viscosity enhancing agent is chitosan or a chitosan derivative, which is present at 0.1-5% w / w, such as 0.1-2.5% w / w, such as 0.5%, 1%, 1.5%, or 2% w / w.

[0070] For example, commercially available low molecular weight HPMC products used as viscosity enhancers in aqueous formulations at 0.5% w / w concentrations have demonstrated viscosities of approximately 35-50 cP, while commercially available high molecular weight HMPC products used at 0.5% w / w concentrations have demonstrated viscosities of approximately 145-200 cP.

[0071] Alternatively, commercially available low molecular weight chitosan products used as viscosity enhancers in aqueous formulations at concentrations of 1-1.5% w / w have demonstrated viscosities ranging from approximately 35-110 cP, while commercially available medium molecular weight chitosan products used at concentrations of 1-1.5% w / w have demonstrated viscosities ranging from approximately 50-175 cP.

[0072] The aqueous formulation may include a viscosity enhancing agent in admixture with one or more additional excipients, such as a preservative or a chelating agent.

[0073] One or more of the optional additional excipients discussed above (i.e., other than peptides and water) are preferably present in the aqueous formulation of the medical product of the invention at individual component concentrations of 10% w / w or less, 9% w / w or less, 8% w / w or less, 7% w / w or less, or 6% w / w or less, preferably 5% w / w or less, 4% w / w or less, 3% w / w or less, or 2% w / w or less, or 1% w / w or less.

[0074] Preferably, the total amount of excipients present in the aqueous formulation of the medical product of the invention is 20% w / w or less, 10% w / w or less, preferably 5% w / w or less, more preferably 4% w / w or less, such as 3% w / w or less or 2% w / w or less.

[0075] Additionally, when any organic excipients are present, the total amount of all such organic excipients in the aqueous formulation of the medical product of the present invention is less than 10% w / w, less than 9% w / w, less than 8% w / w, less than 7% w / w, or less than 6% w / w, preferably less than 5% w / w, less than 4% w / w, less than 3% w / w, or less than 2% w / w, or less than 1% w / w.

[0076] The w / w values ​​are calculated relative to the aqueous formulation excluding the peptide itself, ie relative to the excipients in the formulation.

[0077] Medical Products The medical product of the present invention is a pharmaceutical grade product. The medical product contains the peptide in a ready-to-use form and can be used directly by the end user without the need to add / mix any other components before administering the peptide to a patient in need thereof. In some embodiments, the aqueous formulation is a sterile formulation.

[0078] The medical product is in the form of a sealed container, and in particular, the product of the present invention includes a sterile seal, so that the aqueous formulation in the sealed container is maintained under sterile conditions within the container. The seal is watertight and may be substantially or completely airtight. In the product of the present invention, the integrity of the seal may be maintained for at least 3, 6, 9, 12, 18, 24, or 48 months.

[0079] Preferably, the seal is a manufactured seal, as opposed to a stopper or lid that an individual may apply to a formulation container after forming the formulation from the peptide in solid form, and thus the seal may be considered to be formed by an industrial / mechanical process.

[0080] Aqueous formulations containing peptides are typically administered by injection. Sites of injection include intratumoral, cutaneous, intralesional (e.g., in the case of lentigo carcinoma), subcutaneous, intravenous, and intracavity (e.g., intraperitoneal). The formulations may also be administered in other ways, such as topically, by perfusion or infusion, or transdermally. Intratumoral and intralesional modes of administration are preferred.

[0081] Thus, the medical product of the present invention is preferably a sealed container containing a ready-to-inject aqueous formulation of a peptide, preferably a ready-to-inject aqueous solution of a peptide. In a preferred embodiment, the medical product may be a prefilled syringe or a prefilled syringe cartridge. The prefilled syringe medical product of the present invention may include a needle, or in the case of a cartridge, may require a needle fixed thereto. Such devices are well known in the art and have become common because they allow accurate dosing, reduce waste, and facilitate preparation of injections by users (e.g., clinicians, nurses, or patients themselves) ("Prefilled syringes: An innovation in parenteral packaging," Makwana et al., Int J Pharm Investig. 20122 1(4) pp200-206). Typically, in contrast to syringes containing a formulation mixed for use, a pre-filled syringe (or cartridge) can remain sterile for an indefinite period of time if its integrity is not compromised.

[0082] Prefilled syringe medical products usually include a cap on the needle or tip to maintain a seal on the barrel containing the aqueous formulation, and the other end will typically have a plunger attached to a stopper. The cartridge may be sealed with a luer lock or slip tip type closure. The cartridge may or may not include a plunger, and thus the cartridge may include a dosed and sealed barrel, which is adapted to be fitted into a syringe housing having both a plunger and a needle.

[0083] The barrel containing the aqueous formulation in the prefilled syringe or syringe cartridge may be made of glass or plastic. In some embodiments, the glass may be Pharmaceutical Type 1 glass. In other embodiments, the plastic may be a cyclic olefin polymer, a cyclic olefin copolymer, polypropylene, or polycarbonate.

[0084] In a preferred embodiment, the present invention provides a pre-filled, sealed syringe or syringe cartridge containing an aqueous formulation of LTX-315 having the amino acid sequence of SEQ ID NO. 23, preferably in the form of a pharmaceutically acceptable salt, such as the acetate salt, thereof.

[0085] Other suitable sealed containers include ampoules and vials. Both of these sealed containers are convenient for transporting and storing ready-to-use aqueous formulations of the peptides defined herein. In addition, both are highly suitable for facilitating delivery by injection. Ampoules are typically intended for single use, as their integral cap (e.g., glass) is broken to allow access (e.g., via a needle) to the aqueous formulation therein. Vials may contain larger volumes of aqueous formulations and include a fixed cover or stopper (plug) that can be pierced by a needle (e.g., made of rubber or an equivalent synthetic material). This stopper may conveniently be surrounded by a crimp cap (e.g., made of metal such as aluminum) and optionally covered by a plastic cap. Using a pierceable stopper means that the vial still remains sealed and can contain multiple doses, as the rubber effectively closes the "hole" created by the needle.

[0086] The usable volume of the sealed container may be 0.1-20 ml, e.g., 0.2-10 ml or 0.3-8 ml, preferably 0.5-5 ml. Thus, a sealed container with a usable volume of 2 ml containing an aqueous formulation containing 2 ml of water and 20 mg of peptide would result in a peptide concentration of 10 mg / ml. The amounts of peptide mentioned above refer to net peptide amounts (see the Examples section for an explanation of net peptide amounts).

[0087] The sealed container may contain a single dose and thus constitute a unit dose of peptide, as is the case with a typical prefilled syringe or syringe cartridge. However, the container need not be emptied with a single injection; for example, the same syringe (or cartridge) can deliver multiple injections into a tumor during a single treatment, such as 2-8 injections of 0.2-0.6 ml per injection. When the sealed container is a vial, it may contain multiple doses, such as 2-12 doses, preferably 2-8 doses.

[0088] The sealed container contains an aqueous formulation with a volume of 0.1 to 20.0 ml, preferably 0.2 to 5.0 or 10.0 ml.

[0089] In some embodiments, the sealed container is not completely filled with an aqueous formulation of a peptide as defined herein, and the headspace contains air or an inert gas such as, for example, nitrogen or argon.

[0090] In a further embodiment, the present invention provides a sealed vial or ampoule containing an aqueous formulation of LTX-315 having the amino acid sequence of SEQ ID NO:23, preferably a pharmaceutically acceptable salt, such as the acetate salt, of LTX-315.

[0091] Surprisingly, it has been shown that the medical products can be stored for at least 1, at least 3, at least 4, at least 6, at least 9, at least 12, at least 18, or at least 24 months when stored between manufacture and use at 8° C. or below, such as between 2 and 8° C., such as about 3, 4, 5, 6, or 7° C. When stored in this manner, at least 90%, 91%, 92%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the peptide concentration is maintained.

[0092] Alternatively, the medical product can be stored for at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6 months when stored at about 25° C. and up to 60% relative humidity (RH) between manufacture and use, and when stored in this manner, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the peptide concentration is maintained.

[0093] As shown in the Examples herein, at least 95% of the peptide concentration in the medical product was maintained for at least 24 months when stored at or below 8°C, such as between 2 and 8°C.

[0094] According to the EMA's "ICH Topic Q1A(R2) Stability Testing of New Drug Substances and Products," for drug products intended for refrigerated storage, the minimum period covered by the data at the time of submission is 12 months for long-term conditions (5°C ± 3°C (i.e., 2-8°C)) and 6 months for accelerated conditions (25°C ± 2°C / 60% RH ± 5% RH). The stability results reported in Examples 1, 2, and 4 support a 24-month shelf life at 5°C ± 3°C, with the potential to propose a 36-month shelf life to regulatory authorities by extrapolation of the data.

[0095] Using the ICH definition of "significant change," no significant changes were observed in any of the parameters tested in the stability studies in Examples 1 and 2. In Example 4, using an extrapolation of 4 to 6 months, a slight decrease in purity and a corresponding increase in impurities at storage conditions of 23±2°C would not be considered a "significant change." Thus, according to ICH, extrapolation can be used for any proposed shelf life, and no allowance is required for the effects of short-term deviations from labeled storage conditions, such as during shipping and handling.

[0096] Medical product manufacturing The medical products of the present invention may be manufactured using any conventional method. A peptide in solid form, such as a powder form such as a lyophilized powder, may be dissolved in an aqueous solvent (water, aqueous buffer) under aseptic manufacturing conditions, and the resulting aqueous formulation is preferably sterile filtered and then filled into a container and sealed. The container may be mechanically sealed, for example, by applying a sealant during an industrial process. Alternatively, the peptide may be dissolved in an aqueous solvent (water, aqueous buffer), and the resulting aqueous formulation is then filled into a container and sealed. In a preferred embodiment, the container is autoclaved to obtain a sterile aqueous formulation. When excipients are present in the formulation, depending on the nature and amount of the excipients, these excipients may be mixed or dissolved in the aqueous solvent or in the resulting aqueous formulation.

[0097] Thus, in a further aspect, the present invention provides a method for preparing a medical product in the form of a sealed container containing an aqueous formulation of a peptide as defined herein, wherein the aqueous formulation is introduced into the container, and the filled container is mechanically sealed.In some other embodiments, the present invention provides a method for preparing a medical product in the form of a sealed container containing an aqueous formulation of a peptide as defined herein, wherein the aqueous formulation is prepared as described herein, introduced into the container, and the filled container is mechanically sealed.In a preferred embodiment, the aqueous formulation is sterile filtered before being introduced into the container.In yet another preferred embodiment, the medicinal product is autoclaved.

[0098] In some embodiments, the container is not completely filled with the aqueous formulation, and the headspace contains air or an inert gas, such as nitrogen or argon. Thus, in some embodiments, the present invention provides a method for producing a medical product in the form of a sealed container containing an aqueous formulation of a peptide as defined herein, in which the aqueous formulation is introduced into the container without completely filling the container, and the filled container is mechanically sealed. In some embodiments, the air in the headspace is replaced with an inert gas.

[0099] The preferred features of the medical products of the invention discussed above apply mutatis mutandis to this aspect of the invention.

[0100] Preferred aqueous formulations for use in the medical products of the present invention are provided in the Examples.

[0101] Medical Use of Medical Products The present invention provides methods for treating tumors, both solid and non-solid, by administering an aqueous formulation contained within the medical product described herein to a subject, e.g., a subject in need of treatment. The amount administered should be a therapeutically effective amount, i.e., an amount effective to kill all or a portion of the targeted tumor cells, or to prevent or slow their growth, or to inhibit metastasis or otherwise lessen the deleterious effects of the tumor in the patient. Such a therapeutically effective amount may be administered in one administration, i.e., one dose, or in several administrations, i.e., repeated doses, i.e., a series of doses, over the course of, e.g., several days, weeks, or months, and the clinician should observe an improvement in one or more parameters or symptoms associated with the tumor. The subject is typically a human patient, although non-human animals, e.g., domestic or livestock animals, may also be treated.

[0102] The present invention thus additionally provides a method of treating a tumor in a subject, the method comprising obtaining a medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide comprising: a) consists of a linear sequence of 9 amino acids, b) 5 of those 9 amino acids are cationic and 4 are lipophilic, and c) three of these four lipophilic amino acids are tryptophan and one is non-genetically encoded, and the method further comprises administering to the subject the aqueous formulation contained in a sealed container.

[0103] In some embodiments, the aqueous formulation is administered intratumorally to a subject, i.e., in some embodiments, a method of treating a tumor in a subject comprises obtaining a medical product of the present invention and administering the aqueous formulation contained in the medical product intratumorally to the subject.

[0104] Additionally, disclosed herein is the use of a peptide or a pharmaceutically acceptable salt thereof, wherein the peptide a) consists of a linear sequence of 9 amino acids, b) 5 of those 9 amino acids are cationic and 4 are lipophilic, and c) three of these four lipophilic amino acids are tryptophan and one is non-genetically encoded; The use is for the manufacture of a medical product in the form of a sealed container containing an aqueous formulation of the peptide or a pharmaceutically acceptable salt thereof for treating a tumor in a subject, the aqueous formulation contained in the sealed container being administered to the subject, for example by administration into the tumor of the subject.

[0105] Additionally, disclosed herein is a medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide comprising: a) consists of a linear sequence of 9 amino acids, b) 5 of those 9 amino acids are cationic and 4 are lipophilic, and c) three of the four lipophilic amino acids are tryptophan and one is non-genetically encoded, and the medical product is for use in treating a tumor in a subject, wherein the aqueous formulation contained in a sealed container is administered to the subject, for example, by administration into the tumor of the subject.

[0106] Cancer targets include carcinomas and adenocarcinomas (especially those of the breast, colon, lung, ovary, pancreas, prostate, skin, kidney, and liver (e.g., hepatocellular carcinoma), sarcomas, lymphomas, leukemias, neural cancers (e.g., those of the brain), and melanomas, such as (skin). Cancers of the breast, head and neck, and skin are preferred targets. Melanoma and carcinoma are preferred tumor types for treatment. Tumors for treatment are typically solid tumors and may be metastatic lesions accessible to percutaneous injection.

[0107] The preferred features of the medical products of the invention discussed above apply mutatis mutandis to these aspects of the invention.

[0108] The invention will now be further described with reference to the following non-limiting examples and the following figures. [Brief explanation of the drawings]

[0109] [Figure 1] FIG. 1 graphically presents the results (%) of the assay for all formulations stored at 2-8° C. at T0, T1, and T4, as described in Example 4. [Figure 2] FIG. 1 graphically presents the results (%) of the assay for all formulations stored at 23±2° C. at T0, T1, and T4, as described in Example 4. [Figure 3] FIG. 1 graphically presents the purity results (%) for all formulations stored at 2-8° C. at T0, T1, and T4, as described in Example 4. [Figure 4] FIG. 1 graphically presents the purity results (%) for all formulations stored at 23±2° C. at T0, T1, and T4, as described in Example 4. [Example]

[0110] In all examples, the amount of peptide is the net peptide amount. The net peptide content is the ratio of the peptide of interest to counterions, residual water, and peptidic impurities. The net peptide amount can be calculated by two different methods. 1. The total amount of peptide obtained after synthesis is corrected for its purity (determined by HPLC) and net peptide content (determined by elemental analysis). Net peptide (%) is calculated as follows: Net peptide content (%) x purity (%) 100% The result gives the percentage of net peptide in the total amount of peptide. 2. Net peptide content corresponds to the assay value calculated from analysis by HPLC with external standards. Net peptide (%) is calculated as follows: Apeptide×Cs×100% As×Cpeptide where Apeptide: Peak response of peptide sample As: Peak response of the reference standard Cpeptide: Concentration of peptide sample (mg / mL) Cs: concentration of reference standard (mg / mL) The result gives the percentage of net peptide in the total amount of peptide.

[0111] Example 1.1 Appearance, osmolality, pH, color, and clarity the purpose The objective of this study was to determine whether there were any changes in the appearance, osmolality, pH, color, and clarity of the drug product (LTX-315 dissolved in water for injection (WFI)) contained in sealed vials after storage for different periods of time and under different storage conditions.

[0112] method LTX-315 = acetate salt of peptide with sequence KKWWKKW-Dip-K-NH2 A 20 mg / mL LTX-315 drug product was prepared by dissolving LTX-315 powder in WFI and filtering through a 0.22 μm sterile filter. One milliliter of the drug product was then filled into a vial and sealed. The WFI complied with Ph.Eur. monograph 0169 for WFI. All product-contact vials, glassware, and peripheral equipment were sterilized by rinsing twice with WFI followed by depyrogenation in a hot air oven at 250°C for 120 minutes. Product-contact tubing, stoppers, and overseals were sterilized by rinsing twice with WFI and then autoclaving at 121°C for >15 minutes, followed by drying overnight in a drying oven. Vials were stored as shown in Table 1 below and analyzed by the listed parameters. Appearance was tested visually according to Ph.Eur. 2.9.20, and osmolality was tested according to Ph.Eur. 2.2.35. pH was tested according to Ph.Eur.2.2.3. Color and clarity were tested according to Ph.Eur.2.2.2 and Ph.Eur.2.2.1, respectively.

[0113] Results and Conclusions The drug product was deemed to be of satisfactory appearance, osmolality, pH, color, and clarity at T = 0. There was no change in any of these parameters after 24 months of storage in the dark at 2-8°C in a refrigerator, or after 6 months of storage in the dark at 25°C and 60% relative humidity (RH) in a climate chamber.

[0114] [Table 5]

[0115] Example 1.2 LTX-315 concentration the purpose The objective of this study was to determine whether there were any changes in the concentration of LTX-315 in the drug product (LTX-315 dissolved in WFI) contained in sealed vials after storage for different periods of time and under different storage conditions.

[0116] method The drug product was prepared and stored as described in Example 1.1.

[0117] Reverse phase HPLC using a trifluoroacetic acid (TFA) system and UV detection at 220 nm was used to determine the amount of LTX-315 after storage of vials according to the storage conditions shown in Table 2.

[0118] The column used was RP C18, such as Waters Sunfire, 3.5 μm, 4.6 × 150 mm. The LTX-315 concentration in the test sample solution was 0.5 mg / mL, and the injection volume was 10 μL. The gradient profile used was shown below, and the flow rate was 1.2 mL / min.

[0119] [Table 6]

[0120] Eluent A: Approximately 1800 mL of water, 20 mL of ACN (acetonitrile), and 2 mL of TFA were added to a 2 L volumetric flask. Water was added to volume and mixed thoroughly.

[0121] Eluent B: Approximately 1800 mL of ACN and 2 mL of TFA were added to a 2 L volumetric flask. ACN was added to the volume and mixed thoroughly.

[0122] Results and Conclusions The concentration of LTX-315 after 6 months of storage at 25°C and 60% relative humidity was the same as the concentration at T=0. The slight variation in concentration was due to variability in the analytical method. After 24 months of storage at 2-8°C, there was a very slight decrease in LTX-315 concentration. These results indicated that LTX-315 is stable in aqueous solution under these storage conditions for this period.

[0123] [Table 7]

[0124] Example 1.3 purity the purpose The objective of this study was to determine whether there were any changes in the purity of the drug product (LTX-315 dissolved in WFI) contained in sealed vials after storage for different periods of time and under different storage conditions.

[0125] material and method The drug product was prepared and stored as described in Example 1.1.

[0126] HPLC analysis as described in Example 1.2 was performed to determine the concentration of LTX-315 and the amount of related substances in the drug product, i.e., the amount of substances related to it due to their formation by degradation of LTX-315.

[0127] Results and Conclusions A very slight increase in total related substances was observed for drug products stored at 2-8°C, from 0.88% at T=0 to 1.64% after 24 months, as shown in Table 3. A slight increase in total related substances was also observed for drug products stored at 25°C / 60% RH, from 0.88% at T=0 to 2.31% at T=6 months, as shown in Table 4.

[0128] [Table 8]

[0129] [Table 9]

[0130] conclusion The results of Examples 1.1, 1.2, and 1.3 demonstrated that aqueous formulations of LTX-315 contained in sealed containers as an aqueous solution dissolved in WFI were stable. There was no change in appearance, osmolality, pH, color, or clarity for up to 24 months when the sealed containers were stored at 2-8°C or up to 6 months at 25°C / 60% RH. For medical products, the concentration of the active ingredient (here, LTX-315) should not fall below 90% concentration during the product's shelf life. As shown in Example 1.2, very little change in LTX-315 concentration was observed under these storage conditions and periods, which is also reflected in the little change in related substances shown in Example 1.3.

[0131] [Example 2] Effect of pH the purpose The objective of this study was to investigate the effect of pH on the stability of aqueous formulations of LTX-315 with WFI contained in sealed containers.

[0132] material and method The stability of solutions of LTX-315 dissolved in WFI at a concentration of 20 mg / mL was evaluated at pH 5, pH 6, pH 7, and pH 8. pH adjustment was achieved by using dilute acetic acid (10%) and aqueous sodium hydroxide (0.1 M, 0.5 M, and 1 M NaOH). Formulation details are shown in Table 5, and the testing matrix is ​​shown in Table 6.

[0133] Samples were prepared by dissolving LTX-315 in WFI. Each solution was stirred for 30 minutes to mix, and then the solution was adjusted to the target pH using NaOH and / or acetic acid. When the target pH was achieved, each solution was transferred to an individual 25 mL volumetric flask and brought to volume with WFI. The pH at the final volume was measured, and all solutions were still at target. Each solution was then passed through a 0.2 μm PES membrane syringe filter. The pH of the filtered solutions was measured. All solutions were still at target pH. A 1 mL volume of each solution was filled into a 2 mL clear Pharmaceutical Type 1 glass vial, stoppered, and sealed.

[0134] Samples were incubated under a single stress condition (40°C / 75% RH). Vial contents were then evaluated for appearance, pH, LTX-315 concentration, and purity at T=0, followed by stability evaluation at four additional time points (2, 5, 7, and 14 days). Additional vials were kept in laboratory conditions exposed to ambient light and tested alongside the stress conditions. Control solutions were kept at 2-8°C for the final time point analysis only. No changes in appearance or pH were observed for the control solutions.

[0135] Appearance was assessed by visual inspection as per Ph.Eur.2.9.20, pH was measured as per Ph.Eur.2.2.3 and the concentration and levels of related substances were assessed by HPLC as described above (Example 1.2).

[0136] [Table 10]

[0137] [Table 11]

[0138] result Appearance and pH There was no significant change in appearance or pH of each formulation, regardless of pH, up to 14 days under both ambient and stress (40°C / 75% RH) storage conditions (see Tables 7-10).

[0139] [Table 12]

[0140] [Table 13]

[0141] [Table 14]

[0142] [Table 15]

[0143] concentration The effect of pH on the stability of various LTX-315 formulations was determined by measuring LTX-315 concentrations (assays), and the results are shown in Tables 11–15. Initial assays suggested that LTX-315 concentrations in all formulations were slightly higher (102.2–103.6%) than the target concentration of 20 mg / mL. There was no significant change in concentration for formulations LYT-01, LYT-02, and LYT-03 (pH 5, 6, and 7) over a 14-day period under both ambient and stress conditions. For formulation LYT-04 (pH 8), no change in concentration was observed under ambient storage conditions, whereas a slight decrease in concentration was observed under stress conditions of 40°C / 75% RH.

[0144] [Table 16]

[0145] [Table 17]

[0146] [Table 18]

[0147] [Table 19]

[0148] Purity / Related Substances There was no significant increase in total related substances for formulations LYT-01 (pH 5) and LYT-02 (pH 6) when stored for 14 days at ambient or 40°C / 75% RH conditions (Tables 15-18). For batch LYT-03 (pH 7), there was a small increase in total related substances when stored for 14 days at ambient and especially 40°C / 75% RH conditions (Tables 19 and 20).

[0149] Total related substances for LYT-04 (pH 8) increased when stored at 40°C / 75% RH over 14 days of storage (Tables 21 and 22).

[0150] [Table 20]

[0151] [Table 21]

[0152] [Table 22]

[0153] [Table 23]

[0154] [Table 24]

[0155] [Table 25]

[0156] [Table 26]

[0157] [Table 27]

[0158] conclusion The results of Example 2 showed that LTX-315 was stable in aqueous solution in a sealed container at pH 5 to 7. At pH 8, an increase in related substances was observed, which was also reflected in a decrease in concentration and purity.

[0159] [Example 3] Aqueous formulations for use in medical products of the present invention The aqueous formulation in this Example 3 can be produced using conventional preparation methods, including the following. 1. Weigh out the buffer components and prepare the required amount of aqueous buffer and set aside. 2. Add approximately half the volume of the buffer solution to a vessel equipped with stirring means and optionally heating means, such as a flask and magnetic stirrer, and stir. 3. Add excipients to the buffer solution while stirring. 4. Finally, add the remaining buffer and stir vigorously until the excipients are completely dissolved.

[0160] The peptide may be added in step 2 or step 4. In step 2, the buffer may be heated to about 70°C to facilitate dissolution of certain excipients. When the buffer is heated, the remaining buffer in step 4 is preferably at room temperature, and the peptide is preferably added in step 4. A sterile formulation can be prepared by filtering the formulation through a 0.22 μm sterile filter into a container and sealing the container.

[0161] 3a. Aqueous formulation of LTX-315 at 12 mg / ml containing the viscosity enhancer HPMC (low molecular weight). HPMC was obtained from Safic Alcan, Paris La Defense Cedex, France.

[0162] [Table 28]

[0163] The resulting aqueous formulation had a pH of 5.8, a viscosity of 45 cP, and an osmolality of 380 mmol / L.

[0164] 3b. Aqueous formulation of LTX-315 at 6 mg / ml containing the viscosity enhancer HPMC (high molecular weight). HPMC was obtained from Sufic Alcant, Paris La Défense Cedex, France.

[0165] [Table 29]

[0166] The resulting aqueous formulation had a pH of 5.8, a viscosity of 150 cP, and an osmolality of 380 mmol / L.

[0167] 3c. Aqueous formulation of LTX-315 at 10 mg / ml containing the viscosity enhancer chitosan (low molecular weight). Chitosan was obtained from Chitinor AS, Tromso, Norway.

[0168] [Table 30]

[0169] The resulting aqueous formulation had a pH of 5.6, a viscosity of 60 cP, and an osmolality of 350 mmol / L.

[0170] 3d. Aqueous formulation of LTX-315 at 16 mg / ml containing the viscosity enhancer chitosan (medium molecular weight). Chitosan was obtained from Chitinol AS, Tromsø, Norway.

[0171] [Table 31]

[0172] The resulting aqueous formulation had a pH of 5.6, a viscosity of 120 cP, and an osmolality of 350 mmol / L.

[0173] 3e. A 16 mg / ml aqueous formulation of LTX-302 containing the viscosity enhancer chitosan (medium molecular weight) and the preservative benzyl alcohol.

[0174] [Table 32]

[0175] 3f. 8 mg / ml aqueous formulation of LTX-302 containing the viscosity enhancer chitosan (low molecular weight) and the chelating agent disodium EDTA.

[0176] [Table 33]

[0177] 3g. A 10mg / ml aqueous formulation of LTX-320 containing the antioxidant sodium bisulfite.

[0178] [Table 34]

[0179] 3h. 10mg / ml aqueous LTX-313 formulation containing the stabilizer PEG200.

[0180] [Table 35]

[0181] [Example 4] Stability studies A stability study of LTX-315 in different formulations stored at 23 ± 2°C and 2–8°C was performed by Vitas Analytical Services, Norway. The prepared sterile-filtered formulations were stored in 6R vials, Ph.Eur. Type I (Schott AG), stoppered with lyophilized rubber stoppers V9172 FM257 / 2 ISAF1 Ph.Eur. Type I (Datwyler Pharma Packaging) and capped with flip-off aluminum caps (West Pharmaceutical Services). The formulations were stored at 2–8°C and 23 ± 2°C. Assay, purity, related impurities, appearance, and pH parameters were tested for up to 4 months of storage.

[0182] The formulations are summarized below, with formulations 3-6 containing 10 mg / ml LTX-315 (net peptide).

[0183] [Table 36]

[0184] material Preparation of buffer solutions.

[0185] [Table 37]

[0186] [Table 38]

[0187] The buffer was filtered through a 0.22 μm sterile filter into a flask that had been rinsed with 0.22 μm sterile filtered WFI or into an autoclaved flask.

[0188] Preparation of formulations. LTX-315 was provided in a sealed vial containing 20 mg of sterile, lyophilized peptide in the form of its acetate salt.

[0189] [Table 39]

[0190] [Table 40]

[0191] For Formulations 1 and 2, 2 ml of each sterile buffer solution was injected into the vial containing LTX-315 to dissolve the LTX-315 in the buffer. Two vials were prepared for use in the stability study; one vial was stored at 2-8°C and the other at 23±2°C.

[0192] [Table 41]

[0193] [Table 42]

[0194] [Table 43]

[0195] [Table 44]

[0196] For Formulations 3-6, excipient preparations in acetate buffer were prepared as described in Example 3 and filtered through a 0.22 μm sterile filter. 2 ml of each sterile preparation was injected into a vial containing LTX-315, and LTX-315 was dissolved in the preparation. Two vials were prepared for use in the stability study; one vial was stored at 2-8°C and one was stored at 23±2°C.

[0197] Assay, Purity, and Related Impurities method The analytical method used in the stability study was HPLC-UV with external standard calibration and detection at 220 nm. The formulations were analyzed using a Waters Acquity Cortecs C18+, 1.7 μm, 150 × 3.0 mm ID column at 15 °C with a mobile phase gradient of ACN / HO at 0.612 mL / min and a run time of 38 minutes. The injection volume was 2.0 μL and the injection temperature was 5 °C.

[0198] Purity was calculated by subtracting the measured impurities (total organic impurities) from 100, and related impurities were reported by comparing the diluent chromatogram with the sample chromatogram. Known peaks present in the diluent chromatogram that had the same intensity for both solutions were ignored. This method has been verified to be suitable for the intended purpose.

[0199] The pH was determined according to Ph.Eur.2.2.3.

[0200] Appearance (clarity and color) was determined by visual inspection.

[0201] Results and Conclusions pH and appearance remained essentially unchanged for all formulations at each storage condition after 4 months.

[0202] As presented in Figures 1 and 2, no significant changes were observed in the assay after 4 months of storage at each storage condition for any formulation at either 2-8°C or 23±2°C storage conditions. The small changes observed were due to method variability.

[0203] At the initial time point (T0), no impurities were observed in any of the formulations, and therefore the purity observed for all formulations was 100%. As presented in Figure 3, no significant changes were observed in any of the formulations at 2-8°C during storage for up to 4 months.

[0204] As presented in Figure 4, for formulations stored at 23 ± 2°C, the purity of Formulation 4 was observed to decrease to 98.4% after 1 month (T1), and a further decrease to 97.6% after 4 months (T4) was observed for formulations stored at 23 ± 2°C. This decrease in purity corresponded to a 1.6% and 2.4% increase in total related impurities after 1 and 4 months, respectively.

[0205] For Formulation 3, a decrease in purity to 98.6% was observed after 4 months when stored at 23±2° C., corresponding to an increase in total related impurities to 1.4%.

[0206] conclusion Based on the above, LTX-315 is stable in different aqueous solutions in sealed containers at 2-8°C, as there was essentially no change in any of the parameters tested. Stability results at 23±2°C showed some instability for Formulations 3 and 4, but these are not considered "significant changes" per ICH Q1A (Stability Testing of New Drug Substances and Products).

[0207] Embodiment 1. A medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide comprising: a) consists of a linear sequence of 9 amino acids, b) 5 of the 9 amino acids are cationic and 4 are lipophilic, and c) A medical product in which three of these four lipophilic amino acids are tryptophan and one is non-genetically encoded.

[0208] 2. The medical product of embodiment 1, wherein the cationic amino acid in the peptide is lysine or arginine.

[0209] 3. The medical product of any one of embodiments 1-2, wherein the non-genetically encoded amino acid in the peptide is diphenylalanine or biphenylalanine.

[0210] 4. The medical product of any one of embodiments 1-3, wherein the peptide has an amino acid sequence selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, and 9.

[0211] 5. The medical product of any one of embodiments 1-4, wherein the peptide has an amino acid sequence selected from SEQ ID NOs: 1, 2, and 7.

[0212] 6. The medical product of any one of embodiments 1 to 5, wherein the peptide has an amino acid sequence selected from SEQ ID NOs: 10 to 42.

[0213] 7. The medical product of any one of embodiments 1 to 6, wherein the peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329.

[0214] 8. The medical product of any one of embodiments 1-7, wherein the peptide is in the form of a pharmaceutically acceptable salt.

[0215] 9. The medical product of any one of embodiments 1-8, wherein the peptide is in the form of an acetate salt.

[0216] 10. The medical product of any one of embodiments 1 to 9, wherein the peptide is LTX-315 having the amino acid sequence of SEQ ID NO:23.

[0217] 11. The medical product of any one of embodiments 1 to 10, wherein the peptide is LTX-315 having the amino acid sequence of SEQ ID NO:23, and is in the form of a salt.

[0218] 12. The medical product of any one of embodiments 1 to 11, wherein the peptide is LTX-315 having the amino acid sequence of SEQ ID NO:23 and is in the form of its acetate salt.

[0219] 13. The medical product of any one of embodiments 1 to 12, wherein the peptide is present in the aqueous formulation at a concentration of 1 to 30 mg / ml, preferably 4 to 20 mg / ml, or more preferably 6 to 16 mg / ml, such as for example 10 mg / ml.

[0220] 14. The medical product of embodiment 13, wherein the amount of peptide is a net peptide amount.

[0221] 15. The medical product of any one of embodiments 1-14, wherein the aqueous formulation comprises water for injection.

[0222] 16. The medical product of any one of embodiments 1 to 15, wherein the aqueous formulation consists of the peptide and water for injection.

[0223] 17. The medical product of any one of embodiments 1-15, wherein the aqueous formulation comprises an aqueous buffer such as, for example, an acetate buffer, an aspartate buffer, a sodium benzoate buffer, a benzoic acid buffer, a carbonate buffer, a citrate buffer, a glucono-delta-lactone buffer, a glycine buffer, a glycine HCl buffer, a histidine buffer, a histidine HCl buffer, a hydrobromide buffer, a phosphate buffer (such as PBS), a sodium succinate buffer, a disodium succinate buffer, a succinic acid buffer, a sulfate buffer, a tartrate buffer, and a sodium tartrate buffer, preferably a buffer selected from the group consisting of an acetic acid / acetate buffer, a citrate buffer, and a phosphate buffer.

[0224] 18. The medical product of embodiment 17, wherein the aqueous buffer is a mixture of several different buffers.

[0225] 19. The medical product of any one of embodiments 1-15 and 17-18, wherein the aqueous formulation consists of the peptide and an aqueous buffer.

[0226] 20. The medical product of any one of embodiments 1-15 and 17-18, wherein the aqueous formulation comprises a salt or several salts.

[0227] 21. The medical product of any one of embodiments 1-15, 17-18, and 20, wherein the aqueous formulation further comprises at least one preservative.

[0228] 22. The medical product of embodiment 21, wherein the preservative is selected from the group consisting of benzalkonium chloride, benzyl alcohol, chlorobutanol, metacresol, methylparaben, phenol, potassium sorbate, propylparaben, and thimerosal.

[0229] 23. The medical product of any one of embodiments 1-15, 17-18, and 20-22, wherein the aqueous formulation further comprises at least one chelating agent.

[0230] 24. The medical product of embodiment 23, wherein the chelating agent is selected from the group consisting of calcium disodium EDTA, calcium versetamide sodium, cartelidol, disodium EDTA, DTPA, and sodium EDTA.

[0231] 25. The medical product of any one of embodiments 1-15, 17-18, and 20-24, wherein the aqueous formulation further comprises at least one antioxidant.

[0232] 26. The medical product of embodiment 25, wherein the antioxidant is selected from the group consisting of ascorbic acid, ascorbyl palmitate, citric acid, erythorbic acid, methionine, monothioglycerol, potassium metabisulfite, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium thiosulfate, and thiourea.

[0233] 27. The medical product of any one of embodiments 1-15, 17-18, and 20-26, wherein the aqueous formulation further comprises at least one tonicity adjuster.

[0234] 28. The medical product of embodiment 27, wherein the tonicity adjuster is selected from the group consisting of dextrose, glycerin, lactose, mannitol, potassium chloride, sodium chloride, and sorbitol, perferably sodium chloride.

[0235] 29. The medical product of any one of embodiments 1-15, 17-18, and 20-28, wherein the aqueous formulation further comprises at least one stabilizer.

[0236] 30. The medical product of embodiment 29, wherein the stabilizer is selected from the group consisting of dimethyl sulfoxide, dimethylacetamide, ethanol, glycerol, mannitol, N-methyl-2-pyrrolidone, polyethylene glycol (PEG) 200, PEG 300, PEG 350, PEG 400, PEG 600, propylene glycol, and sorbitol.

[0237] 31. The medical product of any one of embodiments 1-15, 17-18, and 20-28, wherein the aqueous formulation further comprises at least one viscosity enhancing agent.

[0238] 32. The medical product of embodiment 29, wherein the viscosity-enhancing agent is selected from the group consisting of methylcellulose, hydroxypropyl cellulose, hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose, calcium carboxymethylcellulose, sodium carboxymethylcellulose, chitosan, gelatin, hyaluronic acid, maltodextrin, pectin, poloxamer 407, PEG, polyethylene oxide, povidone (PVP), starch such as corn starch and tapioca starch, tragacanth, and beta-cyclodextrin.

[0239] 33. The medical product of embodiment 32, wherein the viscosity enhancing agent is chitosan or a chitosan derivative.

[0240] 34. The medical product of any one of embodiments 1-33, wherein the pH of the aqueous formulation is 5.0-7.4, preferably 5.0-7.0, more preferably 5.0-6.0, such as 6.5.

[0241] 35. The medical product of any one of embodiments 1-34, wherein the viscosity of the aqueous formulation is from 1, 1.5, 2, 2.5, or 3 cP to 300 cP, such as 2.5-250 cP, such as 5-100 cP or 10-200 cP and 20-240 cP, preferably 40-180 cP, such as 45-150 cP or 30-100 cP, more preferably 1.5-10 cP, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 cP, or 10-15 cP or 15-20 cP or 20-35 cP or 35-50 cP or 50-100 cP.

[0242] 36. The medical product of any one of embodiments 1-35, wherein the total amount of excipients present in the aqueous formulation is 20% w / w or less, preferably 5% w / w or less, more preferably 4% w / w or less, such as 3% w / w or less or 2% w / w or less.

[0243] 37. The medical product of any one of embodiments 1-36, wherein the aqueous formulation comprises at least one organic excipient, and the total amount of all such organic excipients is less than 10% w / w, less than 9% w / w, less than 8% w / w, less than 7% w / w, or less than 6% w / w, preferably less than 5% w / w, less than 4% w / w, less than 3% w / w, or less than 2% w / w, or less than 1% w / w.

[0244] 38. The medical product of any one of embodiments 1-37, wherein the aqueous formulation is a sterile formulation.

[0245] 39. The medical product of any one of embodiments 1-38, wherein the aqueous formulation is a ready-to-inject formulation, preferably a ready-to-inject solution of the peptide.

[0246] 40. The medical product of any one of embodiments 1-39, wherein the container is sealed by a manufactured seal.

[0247] 41. The medical product of any one of embodiments 1-40, wherein the container is selected from the group consisting of a prefilled syringe, a syringe cartridge, a vial, and an ampoule.

[0248] 42. The medical product of any one of embodiments 1-41, wherein the usable volume of the sealed container is 0.1-20 ml, such as 0.2-10 ml, such as 0.3-8 ml, preferably 0.5-5 ml.

[0249] 43. The medical product of any one of embodiments 1-42, wherein the sealed container contains the aqueous formulation in a volume of 0.1 to 20 ml, such as 0.2 to 10 ml, preferably 0.2 to 5.0 ml or 10 ml.

[0250] 44. The medical product of any one of embodiments 1-43, wherein the sealed container contains a single dose.

[0251] 45. The medical product of any one of embodiments 1-43, wherein the sealed container contains multiple doses.

[0252] 46. ​​A method of making a medical product according to any one of embodiments 1-45, wherein the aqueous formulation is introduced into a container and the filled container is mechanically sealed.

[0253] 47. A method of treating a tumor in a subject, comprising obtaining a medical product according to any one of embodiments 1 to 45, and administering to the subject an aqueous formulation contained in the medical product.

[0254] 48. The method according to embodiment 47, wherein the aqueous formulation is administered intratumorally to the subject.

[0255] 49. The method according to any one of embodiments 47-48, wherein the tumor is selected from the group consisting of carcinoma (e.g., hepatocellular carcinoma), adenocarcinoma, sarcoma, melanoma, lymphoma, leukemia, germ cell tumor, blastoma, and glioblastoma, and is associated with a cancer selected from the group consisting of neural cancer, brain cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, and head and neck cancer.

[0256] 50. The method according to embodiment 49, wherein the tumor is selected from the group consisting of melanoma, carcinoma, sarcoma, and lymphoma, and is associated with a cancer selected from skin cancer, breast cancer, and head and neck cancer.

Claims

1. 1. A medical product in the form of a sealed container containing an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, said peptide comprising: a) consists of 9 amino acids in a linear sequence, b) 5 of the 9 amino acids are cationic and 4 are lipophilic, and c) A medical product in which three of these four lipophilic amino acids are tryptophan and one is non-genetically encoded.

2. 10. The medical product of claim 1, wherein the cationic amino acid in the peptide is lysine or arginine and the non-genetically encoded amino acid in the peptide is diphenylalanine or biphenylalanine.

3. 3. The medical product of any one of claims 1 to 2, wherein the peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329, preferably in the form of a pharmaceutically acceptable salt, preferably in the form of an acetate salt.

4. The medical product according to any one of claims 1 to 3, wherein said peptide is LTX-315 having the amino acid sequence of SEQ ID NO: 23 and is in the form of a salt, preferably an acetate salt.

5. A medical product according to any one of claims 1 to 4, wherein the peptide is present in the aqueous formulation at a concentration of 1 to 30 mg / ml, preferably 4 to 20 mg / ml.

6. The medical product of any one of claims 1 to 5, wherein the aqueous formulation comprises water for injection or an aqueous buffer solution.

7. The medical product according to any one of claims 1 to 6, wherein the aqueous formulation consists of the peptide and water, preferably water for injection, or of the peptide and an aqueous buffer solution.

8. 8. The medical product of any one of claims 1 to 7, wherein the aqueous formulation further comprises at least one excipient selected from the group consisting of salts, preservatives, tonicity adjusters, stabilizers, antioxidants, viscosity enhancers, and chelating agents.

9. The medical product of any one of claims 1 to 8, wherein the aqueous formulation further comprises at least one viscosity enhancing agent.

10. 10. The medical product of claim 9, wherein the viscosity enhancing agent is chitosan or a chitosan derivative.

11. The medical product of any one of claims 1 to 10, wherein the viscosity of the aqueous formulation is from 1 cP to 300 cP.

12. The medical product according to any one of claims 1 to 11, wherein the aqueous formulation has a pH of 5.0 to 7.

4.

13. A medical product according to any of the preceding claims, wherein the total amount of excipients present in the aqueous formulation is no more than 20% w / w, preferably no more than 5% w / w.

14. 14. The medical product of any of claims 1 to 6 and 8 to 13, wherein the aqueous formulation comprises at least one organic excipient, the total amount of all such organic excipients being less than 10% w / w.

15. The medical product according to any one of claims 1 to 14, wherein said aqueous formulation is a sterile solution of said peptide, preferably a sterile ready-to-inject solution of said peptide.

16. A medical product according to any one of claims 1 to 15, wherein the container is sealed by a manufactured seal.

17. The medical product of any one of claims 1 to 16, wherein the container is selected from the group consisting of a pre-filled syringe, a syringe cartridge, a vial, and an ampoule.

18. A medical product according to any one of the preceding claims, wherein the sealed container contains a volume of the aqueous formulation of 0.1 to 20 ml, preferably 0.2 to 10 ml or 0.2 to 5 ml.

19. The medical product of any one of claims 1 to 18, wherein the sealed container contains a single dose.

20. The medical product of any one of claims 1 to 18, wherein the sealed container contains multiple doses.

21. 21. A method of making a medical product according to any one of claims 1 to 20, wherein the aqueous formulation is introduced into the container and the filled container is mechanically sealed.

22. 21. A method of treating a tumor in a subject, the method comprising obtaining the medical product of any one of claims 1 to 20 and administering to the subject the aqueous formulation contained in the medical product.

23. 23. The method of claim 22, wherein the aqueous formulation is administered into the tumor of the subject.

24. The method of any one of claims 22 to 23, wherein the tumor is selected from the group consisting of carcinoma (e.g., hepatocellular carcinoma), adenocarcinoma, sarcoma, melanoma, lymphoma, leukemia, germ cell tumor, blastoma, and glioblastoma, and is associated with a cancer selected from the group consisting of neural cancer, brain cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, and head and neck cancer.

25. 25. The method of any one of claims 22 to 24, wherein the tumor is selected from the group consisting of melanoma, carcinoma, sarcoma, and lymphoma, and is associated with a cancer selected from skin cancer, breast cancer, and head and neck cancer.

26. Use of a peptide or a pharmaceutically acceptable salt thereof, wherein the peptide is a) consists of 9 amino acids in a linear sequence, b) 5 of the 9 amino acids are cationic and 4 are lipophilic, and c) three of these four lipophilic amino acids are tryptophan and one is non-genetically encoded; The use is for the manufacture of a medical product in the form of a sealed container containing an aqueous formulation of the peptide or a pharmaceutically acceptable salt thereof for treating a tumor in a subject, wherein the aqueous formulation contained in the sealed container is administered to the subject.

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