Medical products comprising aqueous formulations of peptides

Abstract

The present invention relates to a medical product in the form of a sealed container comprising an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, the peptide satisfying: consisting of 9 amino acids arranged linearly; in the nine amino acids, five are cationic amino acids, and four are lipophilic amino acids; and in the four lipophilic amino acids, three are tryptophan, and one is a non-gene coding amino acid. Also provided are methods of making such products wherein an aqueous formulation is introduced into a container and the filled container is mechanically sealed, as well as methods of treatment utilizing such products.

Description

Background Art

[0001] The present invention relates to pharmaceutical products comprising peptides that can be used for cancer treatment. In particular, such products can be manufactured in a ready-to-use dosage form, i.e., for immediate administration to a patient. Thus, such products comprise peptides in a stable formulation that can be transported and stored prior to use.

[0002] The prevalence of cancer in humans and animals, and its contribution to mortality, means that new treatments are continually needed to combat tumors and tumor cells. Eliminating a tumor, reducing its size, destroying its supporting vasculature, or reducing the number of cancer cells circulating in the blood or lymphatic system may be beneficial in many ways, for example, by relieving pain or discomfort, preventing metastasis, facilitating surgical intervention, or prolonging life.

[0003] Cancer therapies designed to target tumors or metastatic cells often rely on cytotoxic activity. This activity can be inherent in the active agent itself or indirectly mediated by the agent, such as by upregulating the host immune response against the tumor. These therapies are more or less selective for the target (tumor or tumor cells) over normal, healthy, or at least non-cancerous, cells and tissues. Less selective therapies can be associated with severe side effects because normal cells are exposed to the cytotoxic activity that the active agent relies on for its therapeutic effect.

[0004] As disclosed in patents WO2010 / 060497, WO2007 / 107748, WO2016 / 091487 and WO2016 / 091490, amphipathic peptides of 9 amino acids comprising a cationic residue and a tryptophan residue are highly effective antitumor agents. These peptides not only produce a direct dissolving effect on tumor cells by disrupting and permeabilizing the cell membrane, but also efficiently attack organelles such as mitochondria and lysosomes and cause them to dissolve. The destruction of organelle membranes can lead to the release of substances with potent immunostimulatory functions, which are commonly referred to as DAMPs (Damage-associated molecular pattern molecules) and include ATP, cytochrome C, mitochondrial CpG DNA sequences, mitochondrial formyl peptides, cathepsins (from lysosomes) and HMGB1 (from the nucleus). Therefore, these lytic peptides stimulate the immune system and provide an adaptive immune response, thereby producing long-term protection against tumor growth. The immune stimulation that occurs when tumor antigens are released by direct lytic action on the tumor leads to enhanced antigen presentation and 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 as a lyophilized powder product in its salt form, and it is recommended to be stored under frozen or refrigerated conditions. LTX-315 is susceptible to hydrolytic degradation, which is usually addressed by preparing the product using non-aqueous solvents or providing it as a dry solid product (for example, by freeze-drying). LTX-315 is provided by commercial suppliers as a dry solid product and needs to be stored below room temperature. For example, LTX-315 is provided by Aldrich Partner Products as a powder product, and the recommended storage conditions are in a dark, inert atmosphere and a refrigerator below -20°C. LTX-315 is provided by BOC Sciences as a lyophilized powder product, and it is recommended to be stored at -20°C. Amadis Chemical also provides LTX-315 as a powder product, and it is recommended to store it in a sealed container in a cool, dry place.

[0006] LTX-315 is typically administered by intratumoral injection, so the powder must be dissolved (i.e., reconstituted) in preparation for administration, for example, in saline to obtain an isotonic solution. Therefore, the administration formulation must be prepared in a clinical setting before use. 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 describes the distribution of a lyophilized powder product that must be reconstituted before use, wherein the lyophilized LTX-315 peptide is said to be dissolved before use.

[0007] In addition, the clinical trial protocols involving LTX-315 conducted in 2021 and 2022 recommend that the lyophilized powder should be reconstituted with saline before on-site use. Preparing injectable drugs (i.e., LTX-315 injection) in a clinical setting (on-site) carries a significantly higher risk of medication errors or microbial contamination than preparing injectable drugs in a ready-to-use form outside of the clinical setting. In addition, the reconstitution process complicates hospital logistics because it makes preparing and administering the product more time-consuming. Therefore, if it is believed that LTX-315 can be safely provided in the form of a ready-to-use aqueous formulation, this approach will be chosen instead of the cumbersome and riskier on-site formulation preparation.

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

[0009] Therefore, there is a need for a convenient, ready-to-use formulation of LTX-315 that can be manufactured, transported, 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 a significant obstacle to the chemical stability of the peptide in aqueous solution. The tryptophan side chain may undergo a large number of chemical reactions in aqueous solution, which are caused by exposure to light, reactive oxygen species, and to some extent pH changes, temperature increases, or exposure to metal cations, which may be extractables / leachables from primary packaging materials such as rubber stoppers and plungers [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, and C.Novel chemical degradation pathways of proteins mediated by tryptophan oxidation:tryptophan side chainfragmentation.J Pharm Pharmacol.2018;70(5):655-665.doi:10.1111 / jphp.12688].

[0011] Short peptides lack the ability to form stable secondary structures in solution. Therefore, the poor physical stability and known chemical degradation pathways of similar short peptides have led to the recognition in the art that relatively short peptides such as LTX-315 are unstable in aqueous solution during the time period from manufacture to administration to patients. In the synthetic peptide octreotide (octreotide, an octapeptide, a modified analog of somatostatin, National Center for Biotechnology Information (2022). PubChem Compound Summary for CID 448601, Octreotide. Retrieved on June 14, 2022, website: https: / / pubchem.ncbi.nlm.nih.gov / compound / Octreotide), a significant chemical degradation pathway involving tryptophan side chains was observed, which may be related to LTX-315. In octreotide, the indole portion of D-tryptophan is transferred to the adjacent lysine (K) side chain through a complex reaction pathway triggered by exposure to UV light. The former D-tryptophan amino acid is then converted to hydroxyglycine, after which the peptide chain is cleaved into two fragments. LTX-315 has one WK motif, in the same order as octreotide, and two KW motifs.

[0012] The octreotide drug product is formulated up to only 1.0 mg / mL and has a pH of approximately 4.0 (isoelectric point of 8.3). This pH is not optimal for intratumoral injection (a pH of 5 to 7 is generally acceptable for intratumoral injection). There are two liquid formulations of octreotide, one for single use and the other for multiple use with a phenolic preservative. Although the product can be stored between 20°C and 30°C (protected from light), it can only be stored for a maximum of 14 days, indicating that the overall stability of this short peptide in aqueous formulations is limited. In addition, octreotide is a cyclic peptide, which generally has improved physical and chemical stability compared to linear peptides.

[0013] Another potential chemical degradation pathway for LTX-315 is the formation of intermolecular ditryptophan crosslinks or bridges through oxidation. The formation of covalent LTX-315 dimers by ditryptophan crosslinks is considered an undesirable degradation product. The potential for ditryptophan formation is also emphasized by comparison with indolicidin. In this 13-residue peptide, a ditryptophan covalent crosslink is characterized between tryptophan residues 6 and 9, which is the same distance as between tryptophan residues 5 and 8 in LTX-315. Although this cross-link was formed under peptide synthesis conditions in TFA, it suggests that this geometry / distance in the sequence allows for the formation of intramolecular Trp-Trp cross-links under appropriate conditions [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].

[0014] A comparison can also be made with degarelix (a decapeptide), which is sold as a lyophilized powder and shows that stability is unstable if formulated into an aqueous solution. Peptides of 9 or 10 amino acids are generally too short to form secondary structures that enhance stability.

[0015] Therefore, it is clear that, due to the presence of multiple tryptophan residues and a length of only 9 amino acids, one skilled in the art would have expected LTX-315 and similar peptides to be susceptible to degradation, particularly in aqueous formulations. This has led to the standard formulation of LTX-315 being a lyophilized powder product, which is recommended to be stored in a refrigerator or freezer to prevent peptide degradation.

[0016] However, the inventors unexpectedly discovered that peptides such as LTX-315 and their pharmaceutically acceptable salts are actually stable in aqueous solutions, meaning that products containing the peptides can be manufactured into ready-to-use dosage forms, i.e., ready for immediate administration directly to a patient. These products contain the peptides in a stable form and can be shipped and stored prior to use. Summary of the Invention

[0017] Therefore, the present invention provides a medicinal product in the form of a sealed container, which comprises an aqueous formulation of a peptide or a pharmaceutically acceptable salt thereof, wherein the peptide satisfies:

[0018] a) consists of 9 linearly arranged amino acids;

[0019] b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and

[0020] c) of the four lipophilic amino acids (i.e., amino acids with a lipophilic R group), three are tryptophan and one is a non-genetically encoded amino acid; and optionally,

[0021] d) the lipophilic residues and cationic residues are arranged in such a way that no more than two of any residue type are adjacent to each other; and further optionally,

[0022] e) The molecule comprises two adjacent pairs of cationic amino acids and one or two adjacent pairs of lipophilic residues.

[0023] Such formulations are suitable for direct administration to the patient, e.g., intratumoral administration, e.g., by injection or by perfusion / infusion. This provides greater convenience for the clinician, who does not need to prepare a formulation for administration to the patient, e.g., by dissolving the peptide in powder form in a suitable solvent.

[0024] peptides

[0025] 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 is positively charged at pH 7.0. Lysine is particularly preferred.

[0026] 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 trimethyllysine and trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-amidinopiperidine-4-carboxylic acid, and 4-guanidinophenylalanine.

[0027] Non-genetically encoded lipophilic amino acids have an R group having at least 7, preferably at least 8 or 9, more preferably at least 10 non-hydrogen atoms. Amino acids having lipophilic R groups are referred to herein as lipophilic amino acids or lipophilic residues. Typically, the lipophilic R group has at least one, preferably two, cyclic groups, which may be fused or linked.

[0028] The lipophilic R group may contain heteroatoms such as O, N or S, but generally no more than one heteroatom, preferably nitrogen. The R group preferably has no more than 2 polar groups, more preferably none or one, most preferably none.

[0029] As disclosed herein, D amino acids, while not strictly genetically encoded, are not considered "non-genetically encoded amino acids" and should be structurally (not just stereospecifically) different from the 20 genetically encoded L amino acids. The molecules disclosed herein may have some or all of the amino acids present in D form.

[0030] The non-genetically encoded lipophilic amino acid preferably has an R group containing no more than 30 non-hydrogen atoms, more preferably has an R group containing no more than 25 non-hydrogen atoms.

[0031] Preferred non-genetically encoded amino acids include: 2-amino-3-(biphenyl-4-yl)propionic acid (biphenylalanine), 2-amino-3,3-diphenylpropionic acid (diphenylalanine), 2-amino-3-(anthracen-9-yl)propionic acid, 2-amino-3-(naphthalen-2-yl)propionic acid, 2-amino-3-(naphthalen-1-yl)propionic acid, 2-amino-3-[1,1':4',1"-terphenyl-4-yl]-propionic acid, 2-amino-3-(2,5,7-tri-tert-butyl-1H-indol-3-yl)propionic acid, ) propionic acid, 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)propionic acid, 2-amino-3-[1,1′:3′,1″-terphenyl-5′-yl]-propionic acid and 2-amino-3-(4-(2,2-diphenylethyl)phenyl)propionic acid. Diphenylalanine and biphenylalanine are particularly preferred.

[0032] In a preferred embodiment, the peptide has one of the following formulas (I) to (V), wherein C represents a cationic amino acid as defined above and L represents a lipophilic amino acid (i.e., tryptophan (Trp / W) or a non-genetically encoded lipophilic amino acid) as defined above. The amino acids are covalently linked via peptide bonds. The free amino or carboxyl termini of these molecules may be modified, preferably the carboxyl terminus is modified to remove the negative charge, most preferably the carboxyl terminus is amidated, and the amide group may be substituted.

[0033] CCLLCCLLC(I) (SEQ ID NO: 1)

[0034] LCCLLCCLC(II) (SEQ ID NO:2)

[0035] CLLCCLLCC(III) (SEQ ID NO:3)

[0036] CCLLCLLCC(IV) (SEQ ID NO:4)

[0037] CLCCLLCCL(V) (SEQ ID NO:5)

[0038] Beta and gamma amino acids as well as alpha amino acids are included in the term "amino acid", as are N-substituted glycines.

[0039] As described above, the peptide comprises a non-genetically encoded lipophilic amino acid. When this residue is represented as L', the preferred peptide is represented by the following formula:

[0040] CCL'LCCLLC(I') (SEQ ID NO:6)

[0041] CCLLCCLL'C(I") (SEQ ID NO:7)

[0042] CCLL'CCLLC(I"')(SEQ ID NO:8)

[0043] LCCLL'CCLC(II') (SEQ ID NO:9)

[0044] Particularly preferred are peptides of formula (I) and (II), of which the peptide of formula (I") is especially preferred.

[0045] The following peptides are most preferred:

[0046]

[0047]

[0048] in:

[0049] Standard single-letter codes are used for genetically encoded amino acids

[0050] Lowercase letters represent D amino acids

[0051] Dip is diphenylalanine

[0052] Bip is biphenylalanine

[0053] Orn is ornithine

[0054] Dap is 2,3-diaminopropionic acid

[0055] Dab is 2,4-diaminobutyric acid

[0056] 1-Nal is 1-naphthylalanine

[0057] 2-Nal is 2-naphthylalanine

[0058] Ath is 2-amino-3-(anthracen-9-yl)propionic acid

[0059] Phe(4,4'Bip) is 2-amino-3-[1,1':4',1"-terphenyl-4-yl]propanoic acid

[0060] 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.

[0061] For use in the present invention, all peptides described herein may be in the form of pharmaceutically acceptable salts. The peptides preferably have a modified, particularly amidated, C-terminus. Suitable pharmaceutically acceptable salts are well known in the art and include salts of inorganic or organic acids, including hydrochlorides, trifluoroacetates, and acetates. Acetate is most preferred.

[0062] Particularly preferred is the peptide known as LTX-315, particularly in the form of a pharmaceutically acceptable salt thereof, preferably the acetate salt thereof.

[0063] Therefore, in a preferred embodiment, the present invention provides a medicinal product in the form of a sealed container, comprising an aqueous preparation of LTX-315 having an amino acid sequence of SEQ ID NO: 23 or a pharmaceutically acceptable salt thereof (preferably an acetate salt thereof).

[0064] Peptide synthesis

[0065] The peptides described herein, such as LTX-315 and pharmaceutically acceptable salts thereof, can be synthesized in any convenient manner by peptide synthesis methods known in the art, preferably by synthesis on a solid support. Synthesis methods are described, for example, in WO2010 / 060497, WO2016 / 091487, and WO2016 / 091490.

[0066] Water-based preparations

[0067] As mentioned above, the medical product of the present invention is in the form of a sealed container comprising an aqueous formulation of a peptide as defined herein.In the following discussion, the term "peptide" refers to the peptide and its pharmaceutically acceptable salts.

[0068] The sealed container may comprise only an aqueous formulation of the peptide as defined herein. In other embodiments, the sealed container is not completely filled with an aqueous formulation of the peptide as defined herein, and the unfilled volume (head space) comprises air or an inert gas, such as nitrogen or argon.

[0069] In some embodiments, the aqueous formulation contains a peptide at a concentration of 1 mg / mL to 30 mg / mL, preferably 4 mg / mL to 20 mg / mL, or more preferably 6 mg / mL to 16 mg / mL. The above amounts of peptide refer to the net amount of peptide (for a description of the net amount of peptide, see the Examples section).

[0070] An aqueous formulation is a formulation in which the solvent used to dissolve the peptide is water, ie, a solution of the peptide in water.

[0071] Since the peptide is readily soluble in water and its aqueous formulation does not need to contain an organic co-solvent for dissolving the peptide, the formulation preferably does not contain an organic co-solvent for dissolving the peptide. In fact, the aqueous formulation may be completely free of organic components (other than the peptide), although organic components may be included, for example, as preservatives, viscosity enhancers, or stabilizers, as described below.

[0072] In some embodiments, the aqueous formulation comprises a solution of the peptide in an aqueous buffer solution, or a mixture of the peptide and the aqueous buffer solution. In some embodiments, the aqueous formulation comprises a solution of the peptide in an aqueous buffer solution, or a mixture of the peptide and the ...

[0073] In some embodiments, the aqueous formulation does not include preservatives and / or stabilizers. In some embodiments, the aqueous formulation does not include hydroxybenzoates and / or EDTA.

[0074] Optionally, the aqueous formulation may contain one or more additional pharmaceutically acceptable ingredients, i.e., excipients, such as those used in medicinal products comprising aqueous formulations of active ingredients, such excipients being well known in the art. Suitable excipients are those used in parenteral formulations, such as formulations administered by intratumoral injection, which is the preferred method of administration in the present invention.

[0075] For example, one or more preservatives may be included in aqueous formulations, particularly when multiple doses are contained in sealed containers. The following table lists examples of preferred preservatives and, if present in the aqueous formulation, their suitable concentration ranges:

[0076] preservative Concentration (w / w) Benzalkonium chloride 0.004% to 0.02% Benzyl alcohol 0.75% to 5% Chlorobutanol 0.25% to 0.5% m-Cresol 0.1% to 0.35% Methylparaben 0.01% to 0.2% phenol 0.15% to 0.5% Potassium sorbate 0.05% to 0.2% Propylparaben 0.005% to 0.035% Thimerosal 0.001% to 0.012%

[0077] Benzyl alcohol is a particularly preferred preservative, preferably present at 0.75% w / w to 2% w / w, preferably 1% w / w. Another preferred preservative is phenol, preferably present at 0.15% w / w to 0.5% w / w.

[0078] One or more chelating agents may be included to chelate metal ions. The following table lists examples of preferred chelating agents. If present in the aqueous formulation, the appropriate concentration range is also listed:

[0079] chelating agents Concentration (w / w) Calcium disodium EDTA 0.01% to 0.1% Calcium versetamide sodium 2% to 3.5% Calteridol 0.01% to 0.03% Disodium EDTA 0.01% to 0.11% DTPA 0.04% to 1.2% Sodium EDTA 0.1% to 0.3%

[0080] EDTA = ethylenediaminetetraacetic acid,

[0081] DTPA = diethylenetriaminepentaacetic acid

[0082] Preferably, the chelating agent is disodium EDTA and / or sodium EDTA.Preferably, the chelating agent (eg disodium EDTA or sodium EDTA) is present at about 0.1% w / w.

[0083] One or more antioxidants may be included. The following table lists examples of preferred antioxidants. If present in the aqueous formulation, their suitable concentration ranges are also listed:

[0084]

[0085]

[0086] A preferred antioxidant is thiourea.

[0087] As mentioned above, in some embodiments, aqueous preparation is the solution of peptide in aqueous buffer or in aqueous buffer mixture. Buffer can be used to maintain the pH of aqueous preparation within the desired range. Preferably, the pH of aqueous preparation is 5.0 to 7.4, more preferably 5.0 to 7.0, even more preferably 5.0 to 6.0 or 6.5. Therefore, particularly preferably following aqueous buffer: acetic acid, acetate, aspartic acid, sodium benzoate, benzoic acid, carbonic acid, citric acid, glucono-δ-lactone, glycine, glycine HCl, histidine, histidine HCl, hydrobromic acid, phosphate (such as PBS), sodium succinate, disodium succinate, succinic acid, sulfuric acid, tartaric acid and / or sodium tartrate. Preferably, aqueous buffer is acetic acid / acetate buffer, citrate buffer and / or phosphate buffer.

[0088] One or more osmotic pressure regulating agents (tonicity modifiers), such as glucose, glycerol, lactose, mannitol, potassium chloride, sodium chloride and / or sorbitol, may be included to obtain an isotonic aqueous formulation, thereby reducing the pain experienced by patients receiving the aqueous formulation by parenteral administration. Preferably, the osmotic pressure regulating agent is sodium chloride.

[0089] 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 include 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 (e.g., PEG stabilizers) can be present in an amount of 1% w / w to 8% w / w, preferably 2% w / w to 7% w / w, for example 3% w / w to 6% w / w.

[0090] The aqueous formulation may also contain one or more viscosity enhancing agents, such as gum arabic, agar, cellulose derivatives (e.g., methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose, carboxymethylcellulose calcium, carboxymethylcellulose sodium), chitosan, gelatin, hyaluronic acid, maltodextrin, pectin, poloxamer 407, PEG, polyethylene oxide, povidone (PVP), starch (e.g., corn starch and tapioca starch), tragacanth gum, and / or β-cyclodextrin.

[0091] The increased viscosity of aqueous formulations (compared to water) can effectively slow the dispersion of peptides from the site of administration. If the formulation is administered intratumorally, the increased viscosity can prolong the time the formulation remains within the tumor without leakage, thereby slowing the dispersion of the peptide via the bloodstream and extending the time the peptide remains in direct contact with tumor cells. This can improve the efficacy of the peptide and / or reduce its toxicity.

[0092] Thus, in some embodiments, the aqueous formulation comprises one or more viscosity enhancers and has a viscosity of from 1.5 cP, 2 cP, 2.5 cP or 3 cP to 300 cP, such as from 2.5 cP to 250 cP, such as from 5 cP to 100 cP or from 10 cP to 200 cP and from 20 cP to 240 cP, preferably from 40 cP to 180 cP, such as from 45 cP to 150 cP or from 30 cP to 100 cP, more preferably from 1.5 cP to 10 cP, such as from 2 cP, 3 cP, 4 cP, 5 cP, 6 cP, 7 cP, 8 cP, 9 cP or 10 cP, or from 10 cP to 15 cP or from 15 cP to 20 cP or from 20 cP to 35 cP or from 35 cP to 50 cP or from 50 cP to 100 cP.

[0093] In some embodiments, wherein the medical product is a prefilled syringe or a prefilled syringe cartridge, and wherein the viscosity of the aqueous formulation of the medical product is greater than 250 cP, greater than 200 cP, greater than 150 cP, or greater than 100 cP, a specialized needle may be used.

[0094] Preferably, the one or more viscosity enhancers are selected from: chitosan or chitosan derivatives or pharmaceutically acceptable salts thereof, such as hydrochloride, hydroxyethyl cellulose, hydroxypropyl cellulose, HPMC, poloxamer 407, PEG and PVP. HPMC and chitosan are particularly preferred viscosity enhancers. Chitosan can be very low molecular weight chitosan (usually <50 kDa), low molecular weight chitosan (usually 50 kDa to 100 kDa), medium molecular weight chitosan (usually 100 kDa to 1000 kDa) or high molecular weight chitosan (usually >1000 kDa). In some embodiments, the chitosan has a degree of deacetylation (DDA) of about 50% to 99%, such as 70% to 99%, such as 80%, preferably about 70% and above.

[0095] Chitosan contains active functional groups that are easily chemically reacted. Through modifications such as acylation, carboxylation, alkylation, and quaternization, chitosan derivatives with properties different from chitosan, such as improved solubility or biological activity, are obtained, but the unique original pharmacological properties of chitosan are retained (see Wang et al. Int. J. Mol. Sci 21(2), 2020, 487, doi: 10.3390 / ijms21020487).

[0096] In some embodiments, the viscosity enhancer is a chitosan derivative, such as O-acylated chitosan and N-acylated chitosan, O-alkylated chitosan and N-alkylated chitosan, chitosan modified by the introduction of hydrophilic groups, such as carboxylic acid groups, quaternary ammonium groups, sulfonic acid groups, phosphoric acid groups, amino groups, ether bonds composed of oxygen groups, hydroxyl groups, carboxylate groups and block polyether groups.

[0097] In some embodiments, the viscosity enhancer is saccharified chitosan, which is obtained by reacting 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-tetraoses, D-pentoses, D-hexoses, D-heptoses, and the like, 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-hamamelis, 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, D-sedoheptulose, and the like. Examples of such oligosaccharides include fructo-oligosaccharide (FOS), galacto-oligosaccharide (GOS), mannan-oligosaccharide (MOS), etc. An example of saccharified chitosan is galacto-chitosan. Saccharified chitosan and its synthesis are described in, for example, US 5747475A and WO 2002 / 040055 A2 and WO 2013 / 109732 A2, the contents of which are incorporated herein by reference.

[0098] In some embodiments, the viscosity enhancer is a saccharified chitosan having a glycosylation percentage of about 0.1% to about 90% on its native free amino groups, for example, about 0.1% to about 30%, or for example, about 2% to 15%, for example, about 12.5%. In some embodiments, the saccharified chitosan has a molecular weight of about 50 kD to about 2000 kD, for example, about 50 kD to about 1500 kD, for example, about 250 kD or 300 kD. In some embodiments, the saccharified chitosan has a DDA of about 50% to 99%, for example, 70% to 99%, for example, 80%. In some embodiments, the saccharified chitosan has a DDA of about 50% to 99% (e.g., 70% to 99%, for example, 80%), a percentage glycosylation of about 0.1% to about 90% (e.g., about 0.1% to about 30% or for example, about 2% to 15%, for example, about 12.5%) acting on its native free amino groups, and has a molecular weight of about 50 kD to about 2000 kD (e.g., about 50 kD to about 1500 kD, for example, about 250 kD or about 300 kD).

[0099] The viscosity enhancer is preferably present at 0.1% w / w to 5% w / w, such as 0.15% w / w to 2.5% w / w or 0.2% w / w to 2% w / w or 0.25% w / w to 1.5% w / w or 0.3% w / w to 1% w / w or 0.35% w / w to 0.9% w / w. In some embodiments, the viscosity enhancer is chitosan or a chitosan derivative, which is present at 0.1% w / w to 5% w / w, such as 0.1% w / w to 2.5% w / w, such as 0.5% w / w, 1% w / w, 1.5% w / w or 2% w / w.

[0100] For example, using a commercially available low molecular weight HPMC product as a viscosity enhancer, a viscosity of approximately 35 cP to 50 cP has been demonstrated at a concentration of 0.5% w / w in an aqueous formulation, while using a commercially available high molecular weight HMPC product, a viscosity of approximately 145 cP to 200 cP has been demonstrated at a concentration of 0.5% w / w.

[0101] Alternatively, the use of commercially available low molecular weight chitosan products as viscosity enhancers has demonstrated viscosities ranging from about 35 cP to about 110 cP at concentrations of 1% w / w to 1.5% w / w in aqueous formulations, while the use of commercially available medium molecular weight chitosan products has demonstrated viscosities ranging from about 50 cP to 175 cP at concentrations of 1% w / w to 1.5% w / w.

[0102] Aqueous formulations may contain a viscosity enhancing agent mixed with one or more additional excipients, such as preservatives or chelating agents.

[0103] The one or more optional additional excipients described above (i.e., other than the peptide and water) are preferably present in the aqueous formulation of the medicinal product of the present invention at a concentration of no more than 10% w / w, no more than 9% w / w, no more than 8% w / w, no more than 7% w / w or no more than 6% w / w of the individual components, preferably no more than 5% w / w, no more than 4% w / w, no more than 3% w / w, no more than 2% w / w or no more than 1% w / w.

[0104] Preferably, the total amount of excipients present in the aqueous formulation in the medicinal product of the present invention does not exceed 20% w / w, does not exceed 10% w / w, preferably does not exceed 5% w / w, more preferably does not exceed 4% w / w, for example does not exceed 3% w / w or does not exceed 2% w / w.

[0105] Furthermore, if any organic excipients are present, the total amount of all such organic excipients in the aqueous formulation of the medicinal 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.

[0106] The w / w values ​​are calculated for the aqueous formulation excluding the peptide itself (ie for the excipients in the formulation).

[0107] Medical products

[0108] The medical product of the present invention is a pharmaceutical grade product. The medical product comprises a ready-to-use peptide that can be used directly by the end user without the need to add / mix any other ingredients before administration to a patient in need thereof. In some embodiments, the aqueous formulation is a sterile formulation.

[0109] The medical product is in the form of a sealed container, particularly a product of the present invention comprising a sterile seal, such that the aqueous formulation within the sealed container is maintained under sterile conditions within the container. The seal is waterproof and can be substantially or completely airtight. In the product of the present invention, the integrity of the seal can be maintained and can be maintained for at least 3, 6, 9, 12, 18, 24, or 48 months.

[0110] Preferably, the seal is a manufactured seal; this is different from a stopper or cap which can be applied to the container of the formulation by an individual after the formulation has been produced from the solid form of the peptide. Thus, the seal can be considered to be formed in an industrial / mechanical process.

[0111] Aqueous formulations containing peptides are typically administered by injection. Injection sites include intratumoral, dermal, intralesional (e.g., in the case of cancerous nevi), subcutaneous, intravenous, intracavitary (e.g., intraperitoneal). The formulation can be administered in other ways, such as topically, by perfusion or infusion, or transdermally. Intratumoral and intralesional administration are preferred.

[0112] Therefore, the medical product of the present invention is preferably a sealed container containing a ready-to-use injection peptide aqueous formulation, preferably a ready-to-use injection peptide aqueous solution. In a preferred embodiment, it is 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, a needle needs to be fixed thereon. Such devices are well known in the art and are increasingly popular because they enable accurate dosing, reduce waste, and are easy for users (e.g., clinicians, nurses, or patients themselves) to prepare injections (Prefilled syringes: An innovation in parenteral packaging, Makwana et al, Int J Pharm Investig. 20122 1 (4) pp200-206). Generally, compared to mixed preparations in syringes, prefilled syringes (or cartridges) can remain sterile indefinitely, provided that their integrity is not compromised.

[0113] Prefilled syringe medical products typically have a cap at the needle or tip to keep the barrel containing the aqueous formulation sealed; the other end usually has a plunger connected to a stopper. Cartridges can be sealed with a Luerlock or slip-tip type closure. Cartridges may or may not contain a plunger, so they can contain a metered, sealed barrel that fits into a syringe housing with a plunger and needle.

[0114] The barrel containing the aqueous formulation in the prefilled syringe or syringe cartridge can be made of glass or plastic. In some embodiments, the glass can be pharmaceutical type 1 glass. In other embodiments, the plastic can be cyclic olefin polymer, cyclic olefin copolymer, polypropylene, or polycarbonate.

[0115] In a preferred embodiment, the present invention provides a pre-filled and sealed syringe or syringe cartridge comprising an aqueous formulation of LTX-315 having an amino acid sequence of SEQ ID NO: 23, preferably in the form of a pharmaceutically acceptable salt, such as an acetate salt thereof.

[0116] Other suitable sealed containers include ampoules and vials. Both of these sealed containers are convenient ways to transport and store the instant aqueous peptide formulations defined herein. Both are very suitable for promoting delivery by injection. Ampoules are usually disposable because the one-piece lid (e.g., glass) is broken open to obtain (e.g., by a needle) the aqueous formulation therein. Vials can contain aqueous formulations of larger capacity and contain a fixed cap or bottle stopper (bucket) that can be pierced with a needle (e.g., made of rubber or equivalent synthetic materials). The bottle stopper can be conveniently surrounded by a crimped lid (e.g., made of a metal such as aluminum) and optionally covered with a plastic cap. Using a penetrable bottle stopper means that the vial can accommodate multiple doses, and the vial remains sealed because the rubber effectively closes the "hole" produced by the needle.

[0117] The usable volume of the sealed container can be 0.1 mL to 20 mL, for example 0.2 mL to 10 mL or 0.3 mL to 8 mL, preferably 0.5 mL to 5 mL. Thus, a sealed container with a usable volume of 2 mL, containing an aqueous formulation consisting of 2 mL of water and 20 mg of peptide, has a peptide concentration of 10 mg / mL. The above amount of peptide refers to the net amount of peptide (see the Examples section for a description of the net amount of peptide).

[0118] The sealed container can contain a single dose, thereby constituting a unit dose of the peptide, as is typically the case with prefilled syringes or syringe cartridges. However, the container may not be emptied during a single injection, for example, the same syringe (or cartridge) can be used to administer multiple injections to the tumor during a single treatment, for example, 2 to 8 injections of 0.2 mL to 0.6 mL per injection. If the sealed container is a vial, it can contain multiple doses, for example 2 to 12 doses, preferably 2 to 8 doses.

[0119] The volume of the aqueous formulation contained in the sealed container is 0.1 mL to 20.0 mL, preferably 0.2 mL to 5.0 mL or 10.0 mL.

[0120] In some embodiments, the sealed container is not completely filled with the aqueous formulation of a peptide as defined herein, and the headspace comprises air or an inert gas, such as nitrogen or argon.

[0121] In another embodiment, the present invention provides a sealed vial or ampoule comprising an aqueous formulation of LTX-315 having an amino acid sequence of SEQ ID NO: 23 (preferably a pharmaceutically acceptable salt of LTX-315, such as acetate).

[0122] Unexpectedly, the results show that when stored at 8°C or below (e.g., 2°C to 8°C, such as about 3°C, 4°C, 5°C, 6°C, or 7°C), the medicinal product can be stored for at least 1 month, at least 3 months, at least 4 months, at least 6 months, at least 9 months, at least 12 months, at least 18 months, or at least 24 months between manufacture and use. When stored in this manner, at least 90%, 91%, 92%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the peptide concentration is maintained.

[0123] Alternatively, the medicinal product can be stored for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months between manufacture and use when stored at room temperature at about 25° C. and a relative humidity (RH) of up to 60%. When stored in this manner, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the peptide concentration can be maintained.

[0124] As shown in the Examples herein, at least 95% of the peptide concentration in the medicinal product is maintained for at least 24 months when stored at 8°C or below (eg, 2°C to 8°C).

[0125] According to EMA's "ICH Topic Q1A(R2) Stability Testing of new Drug Substances and Products," for drug products intended to be stored in a refrigerator, the minimum time covered by the submitted data for long-term conditions (5°C ± 3°C (i.e., 2°C to 8°C)) is 12 months, and the minimum time covered by accelerated conditions (25°C ± 2°C / 60% RH ± 5% RH) is 6 months. The stability results reported in Examples 1, 2, and 4 support a shelf life of 24 months at 5°C ± 3°C, and by extrapolation of the data, it is possible to propose a shelf life of 36 months to regulatory authorities.

[0126] Using the ICH definition of "significant change," no significant changes were observed in any of the test parameters in the stability studies of Examples 1 and 2. In Example 4, using the extrapolation approach from 4 to 6 months, the minor decrease in purity and corresponding increase in impurities at 23°C ± 2°C storage conditions was not considered a "significant change." Therefore, according to ICH, extrapolation can be used for any proposed shelf life, and the effects of short-term deviations from label storage conditions (e.g., during shipping and handling) do not need to be considered.

[0127] Manufacturing of medical products

[0128] The medical product of the present invention can be manufactured by any conventional method. The peptide in solid form (e.g., powder form, such as lyophilized powder) can be dissolved in an aqueous solvent (water, aqueous buffer) under aseptic manufacturing conditions, and the resulting aqueous preparation is preferably aseptically filtered, then filled into a container and sealed. The container can be mechanically sealed, such as by applying a seal in an industrial process. Alternatively, the peptide can be dissolved in an aqueous solvent (water, aqueous buffer), and then the resulting aqueous preparation is filled into a container and sealed. In a preferred embodiment, the container is autoclaved to obtain a sterile aqueous preparation. If there is an excipient in the preparation, then, according to the nature and amount of the excipient, it is mixed or dissolved in an aqueous solvent, or mixed or dissolved in the resulting aqueous preparation.

[0129] Thus, in another aspect, the present invention provides a method for manufacturing a medical product in the form of a sealed container, the medical product comprising 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 other embodiments, the present invention provides a method for manufacturing a medical product in the form of a sealed container, the medical product comprising 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 prior to introduction into the container. In another preferred embodiment, the medical product is autoclaved.

[0130] In some embodiments, the container is not completely filled with the aqueous formulation and the headspace comprises air or an inert gas, such as nitrogen or argon. Thus, in some embodiments, the present invention provides a method for manufacturing a medical product in the form of a sealed container comprising an aqueous formulation of a peptide as defined herein, wherein the aqueous formulation is introduced into the container but not completely filled, and the filled container is mechanically sealed. In some embodiments, the air in the headspace is replaced by an inert gas.

[0131] The preferred features of the medical product of the invention described above apply mutatis mutandis to this aspect of the invention.

[0132] Preferred aqueous formulations for use in the medicinal products of the present invention are listed in the Examples.

[0133] Medical uses of medical products

[0134] The present invention provides a method for treating tumors (solid tumors and non-solid tumors) by administering an aqueous formulation contained in a medical product described herein to a subject (e.g., a subject in need thereof). The amount administered should be a therapeutically effective amount, i.e., effectively killing all or part of the target tumor cells or preventing or reducing their proliferation rate, or inhibiting metastasis or alleviating the deleterious effects of the tumor on the patient. This therapeutically effective amount can be administered once, i.e., a single dose, or in fractions, i.e., repeated doses, i.e., a series of doses, such as over the course of several days, weeks, or months, the clinician should observe an improvement in one or more parameters or symptoms associated with the tumor. The subject is typically a human patient, but non-human animals, such as domestic animals or livestock, can also be treated.

[0135] Therefore, the present invention also provides a method for treating a tumor in a subject, the method comprising obtaining a medical product in the form of a sealed container comprising an aqueous preparation of a peptide or a pharmaceutically acceptable salt thereof, and administering the aqueous preparation contained therein to the subject, wherein the peptide satisfies: a) consisting of 9 amino acids arranged linearly; b) of the 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and c) of the 4 lipophilic amino acids, 3 are tryptophan and 1 is a non-genetically encoded amino acid.

[0136] In some embodiments, the aqueous formulation is administered to a tumor in a subject, i.e., in some embodiments, the method of treating a tumor in a subject comprises obtaining a medical product of the invention and administering the aqueous formulation contained therein to the tumor in the subject.

[0137] The present invention also discloses the use of a peptide or a pharmaceutically acceptable salt thereof for preparing a medical product in the form of a sealed container containing an aqueous preparation of the peptide or a pharmaceutically acceptable salt thereof for treating a tumor in a subject, wherein the peptide satisfies the following conditions: a) consisting of 9 amino acids arranged linearly; b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and c) of these 4 lipophilic amino acids, 3 are tryptophan and 1 is a non-genetically encoded amino acid, wherein the aqueous preparation contained in the sealed container is administered to a subject, for example, into a tumor of the subject.

[0138] Also 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 for treating a tumor in a subject, wherein the peptide satisfies the following conditions: a) consisting of 9 amino acids arranged linearly; b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and c) of these 4 lipophilic amino acids, 3 are tryptophan and 1 is a non-genetically encoded amino acid, wherein the aqueous formulation contained in the sealed container is administered to a subject, for example, into a tumor of the subject.

[0139] Cancer targets include carcinomas and adenocarcinomas (particularly from the breast, colon, lung, ovary, pancreas, prostate, skin, kidney and liver (e.g., hepatocellular carcinoma)), sarcomas, lymphomas, leukemias, nervous system cancers (e.g., from the brain) and melanomas (e.g., skin). Breast cancer, head and neck cancer, and skin cancer are preferred targets. Melanoma and carcinoma are preferred tumor types for treatment. Tumors for treatment are typically solid tumors and may be metastatic lesions that can be treated by transdermal injection.

[0140] The preferred features of the medical product of the invention described above apply mutatis mutandis to these aspects of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0141] The present invention will now be further described with reference to the following non-limiting examples and figures:

[0142] Figure 1 : Graphical representation of the assay results (%) for each formulation stored at 2°C to 8°C at T0, T1 and T4, as described in Example 4.

[0143] Figure 2 : Graphical representation of the assay results (%) for each formulation stored at 23°C ± 2°C at T0, T1 and T4, as described in Example 4.

[0144] Figure 3 : Graphical representation of the purity results (%) for each formulation stored at 2°C to 8°C at T0, T1 and T4, as described in Example 4.

[0145] Figure 4 : Graphical representation of the purity results (%) for each formulation stored at 23°C ± 2°C at T0, T1 and T4, as described in Example 4. DETAILED DESCRIPTION

[0146] In all examples, the amount of peptide is the net peptide amount. Net peptide content is the fraction of the peptide in question relative to counterions, residual water, and peptide impurities. Net peptide amount can be calculated using two different methods:

[0147] 1. The total peptide obtained after synthesis was calibrated for purity (determined by HPLC) and net peptide content (determined by elemental analysis). Net peptide (in %) was calculated as follows:

[0148]

[0149] The results are given as a percentage of the net peptides in the total peptides.

[0150] 2. Net peptide content corresponds to the measured value calculated by external standard HPLC analysis. Net peptide (in %) is calculated as follows:

[0151]

[0152] in:

[0153] Peptide A: Peak response of peptide sample

[0154] As: Peak response of the reference standard

[0155] C-peptide: Concentration of peptide sample (mg / mL)

[0156] Cs: Concentration of reference standard (mg / mL)

[0157] The results are given as a percentage of the net peptides in the total peptides.

[0158] Example 1.1: Appearance, Osmolality, pH, Color, and Clarity

[0159] Purpose

[0160] The objective of this study was to determine whether the drug product (LTX-315 dissolved in water for injection (WFI)) in sealed vials undergoes any changes in appearance, osmolarity, pH, color, and clarity after storage for various time periods and under various storage conditions.

[0161] method

[0162] LTX-315 = acetate salt of a peptide with the sequence KKWWKKW-Dip-K-NH2

[0163] LTX-315 powder was dissolved in WFI and filtered through a 0.22 μm sterile filter to prepare a 20 mg / mL LTX-315 drug product. 1 mL of drug product was then filled into vials and sealed. The WFI conforms to the WFI specifications in Ph. Eur. Monograph 0169. All product-contact vials, glassware, and peripheral equipment were rinsed twice with WFI and sterilized by depyrogenation in a hot air oven at 250°C for 120 minutes. Product-contact tubes, stoppers, and outer seals were rinsed twice with WFI and then autoclaved at 121°C for >15 minutes and then dried in a desiccator overnight. Vials were stored as listed in Table 1 below and analyzed according to the parameters listed. Appearance was visually tested 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.

[0164] Results and Conclusions

[0165] The drug product's appearance, osmolarity, pH, color, and clarity were all considered satisfactory at T = 0. There was no change in these parameters after 24 months of storage in a refrigerator at 2°C to 8°C in the dark or 6 months of storage in a climate chamber at 25°C, 60% relative humidity (RH) in the dark.

[0166] Table 1: Appearance, osmolarity, pH, color, and clarity of the solution

[0167]

[0168]

[0169] B9 and 1 NTU are reference standards described in Ph. Eur. 2.2.1 and Ph. Eur. 2.2.2.

[0170] Example 1.2: LTX-315 concentration

[0171] Purpose

[0172] The objective of this study was to determine whether there were any changes in the concentration of LTX-315 after storage of the drug product (LTX-315 dissolved in WFI) in sealed vials for various periods of time and under various storage conditions.

[0173] method

[0174] The drug product was prepared and stored as described in Example 1.1.

[0175] The amount of LTX-315 after storage of the vials according to the storage conditions listed in Table 2 was determined using a trifluoracetic acid (TFA) system and reverse phase HPLC with UV detection at 220 nm.

[0176] The chromatographic column used was an RP C18 column, such as a Waters Sunfire, 3.5 μm, 4.6 x 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.

[0177] Gradient distribution

[0178] Eluent A: Add approximately 1800 mL of water, 20 mL of ACN (acetonitrile), and 2 mL of TFA to a 2 L volumetric flask. Add water to the mark and mix thoroughly.

[0179] Eluent B: Add approximately 1800 mL of ACN and 2 mL of TFA to a 2 L volumetric flask. Fill to volume with ACN and mix thoroughly.

[0180] Results and Conclusions

[0181] After six months of storage at 25°C and 60% relative humidity, the concentration of LTX-315 was the same as at T = 0. The slight change in concentration is due to variability in the analytical method. After 24 months of storage at 2°C to 8°C, the LTX-315 concentration decreased slightly. These results indicate that LTX-315 is stable in aqueous solution over this period of time under these storage conditions.

[0182] Table 2:

[0183]

[0184]

[0185] 1 Calculated as a percentage of the theoretical value of 20 mg / mL

[0186] 2 % recovery compared to pre-filtration results

[0187] Example 1.3: Purity

[0188] Purpose

[0189] The objective of this study was to determine whether there is any change in the purity of the drug product (LTX-315 dissolved in WFI) in sealed vials after storage for various periods of time and under various storage conditions.

[0190] Materials and methods

[0191] The drug product was prepared and stored as described in Example 1.1.

[0192] HPLC analysis was performed as described in Example 1.2 to determine the concentration of LTX-315 and the amount of related substances, ie substances formed due to degradation of and therefore related to LTX-315 in the drug product.

[0193] Results and Conclusions

[0194] For drug products stored at 2°C to 8°C, a slight increase in total related substances was observed, 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.

[0195] Table 3: Purity / related substances (area %), 2°C to 8°C

[0196]

[0197]

[0198] 1 Sum of related substances > 0.05% LOQ = Limit of Quantitation

[0199] Table 4: Purity / related substances (area %), 25℃ / 60%RH

[0200]

[0201] 1 The sum of related substances> 0.05%

[0202] LOQ = limit of quantitation

[0203] in conclusion

[0204] The results of Examples 1.1, 1.2, and 1.3 demonstrate that an aqueous formulation of LTX-315 dissolved in WFI and packaged in a sealed container as an aqueous solution is stable; when stored in a sealed container at 2°C to 8°C for up to 24 months, or at 25°C / 60% RH for up to 6 months, there is no change in appearance, osmolarity, pH, color, or clarity. For medical products, the concentration of the active ingredient (here, LTX-315) must not fall below 90% of the concentration expected during the shelf life of the product. As shown in Example 1.2, the measured LTX-315 concentration showed little change over these storage conditions and time periods, which is also reflected in the minimal changes in related substances, as shown in Example 1.3.

[0205] Example 2: Effect of pH

[0206] Purpose

[0207] The objective of this study was to investigate the effect of pH on the stability of aqueous formulations of LTX-315 in WFI in sealed containers.

[0208] Materials and methods

[0209] The stability of LTX-315 solutions 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 performed using dilute acetic acid (10%) and aqueous sodium hydroxide solutions (0.1 M, 0.5 M, and 1 M NaOH). Formulation details are shown in Table 5, and the test matrix is ​​shown in Table 6.

[0210] Samples were prepared by dissolving LTX-315 in WFI, stirring each solution for 30 minutes to mix, and then adjusting the solution to the target pH using NaOH and / or acetic acid. After reaching the target pH, each solution was transferred to a separate 25 mL volumetric flask and brought to volume with WFI. The pH of the final volume was measured, and all solutions remained at the target value. Each solution was then passed through a 0.2 μm PES membrane syringe filter. The pH of the filtered solution was measured; all solutions remained at the target pH. Each solution was filled into a clear, pharmaceutical-grade Type 1 glass 2 mL vial at a volume of 1 mL, stoppered, and sealed.

[0211] The samples were incubated under a single stress condition (40°C / 75% RH). The vial contents were then evaluated for appearance, pH, LTX-315 concentration, and purity at T=0, and then stability was evaluated at four additional time points (2, 5, 7, and 14 days). Additional vials were placed under laboratory conditions at room temperature and light and tested in conjunction with the stress condition. A control solution was analyzed at 2°C to 8°C only at the final time point. No changes in appearance or pH were observed for the control solution.

[0212] Appearance was assessed by visual inspection according to Ph. Eur. 2.9.20. pH was determined according to Ph. Eur. 2.2.3. Concentrations and levels of related substances were assessed by HPLC as described above (Example 1.2).

[0213] Table 5: preparation

[0214]

[0215]

[0216] Table 6: Test matrix

[0217]

[0218] RS: Related substances

[0219] result

[0220] Appearance and pH

[0221] Regardless of pH, there were no significant changes in the appearance or pH of each formulation under both room temperature and stressed (40°C / 75% RH) storage conditions for up to 14 days (see Tables 7 to 10).

[0222] Table 7: Appearance and pH, LYT-01

[0223]

[0224] Table 8: Appearance and pH, LYT-02

[0225]

[0226]

[0227] Table 9: Appearance and pH, LYT-03

[0228]

[0229] Table 10: Appearance and pH, LYT-04

[0230]

[0231] concentration

[0232] 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 to 15. Preliminary measurements showed that LTX-315 concentrations in all formulations were slightly above the target concentration of 20 mg / mL (102.2% to 103.6%). The concentrations of LYT-01, LYT-02, and LYT-03 formulations (pH 5, 6, and 7, respectively) did not change significantly over a 14-day period at room temperature and under stress conditions. For formulation LYT-04 (pH 8), no change in concentration was observed under room temperature storage conditions, while a slight decrease in concentration was observed under stress conditions at 40°C / 75% RH.

[0233] Table 11: Determination, LYT-01

[0234]

[0235] 1 Percentage of the theoretical value of 20 mg / mL

[0236] 2 Percentage of theoretical value of pre-filtration determination

[0237] 3 Percentage of initial (T=0) results

[0238] Table 12: Determination of LYT-02

[0239]

[0240] 1 Percentage of the theoretical value of 20 mg / mL

[0241] 2 Percentage of theoretical value of pre-filtration determination

[0242] 3 Percentage of initial (T=0) results

[0243] Table 13: Determination of LYT-03

[0244]

[0245] 1Percentage of the theoretical value of 20 mg / mL

[0246] 2 Percentage of theoretical value of pre-filtration determination

[0247] 3 Percentage of initial (T=0) results

[0248] Table 14: Determination of LYT-04

[0249]

[0250] 1 Percentage of the theoretical value of 20 mg / mL

[0251] 2 Percentage of theoretical value of pre-filtration determination

[0252] 3 Percentage of initial (T=0) results

[0253] Purity / Related Substances

[0254] No significant increase in the total amount of related substances was observed for formulations LYT-01 (pH 5) and LYT-02 (pH 6) when stored at room temperature or 40°C / 75% RH for 14 days (Tables 15 to 18). A slight increase in the total amount of related substances was observed for the LYT-03 batch (pH 7) when stored at room temperature, particularly after 14 days at 40°C / 75% RH (Tables 19 and 20).

[0255] When stored at 40°C / 75%RH for 14 days, the total amount of related substances in LYT-04 (pH 8) increased (Tables 21 and 22).

[0256] Table 15: Purity / related substances (area %), LYT-01 room temperature

[0257]

[0258]

[0259] 1 The sum of related substances> 0.05%

[0260] ND = Not Detected

[0261] LOQ = limit of quantitation

[0262] Table 16: Purity / related substances (area %), LYT-01 40℃ / 75%RH

[0263]

[0264]

[0265] 1 The sum of related substances> 0.05%

[0266] ND = Not Detected

[0267] LOQ = limit of quantitation

[0268] Table 17: Purity / related substances (area %), LYT-02 room temperature

[0269]

[0270]

[0271] 1 The sum of related substances> 0.05%

[0272] ND = Not Detected

[0273] LOQ = limit of quantitation

[0274] Table 18: Purity / related substances (area %), LYT-02 40℃ / 75%RH

[0275]

[0276]

[0277] 1 The sum of related substances> 0.05%

[0278] ND = Not Detected

[0279] LOQ = limit of quantitation

[0280] Table 19: Purity / related substances (area %), LYT-03 room temperature

[0281]

[0282]

[0283] 1 The sum of related substances> 0.05%

[0284] ND = Not Detected

[0285] LOQ = limit of quantitation

[0286] Table 20:Purity / related substances (area %), LYT-03 40℃ / 75%RH

[0287]

[0288]

[0289] 1 The sum of related substances> 0.05%

[0290] ND = Not Detected

[0291] LOQ = limit of quantitation

[0292] Table 21: Purity / related substances (area %), LYT-04 room temperature

[0293]

[0294]

[0295] 1 The sum of related substances> 0.05%

[0296] ND = Not Detected

[0297] LOQ = limit of quantitation

[0298] Table 22: Purity / related substances (area %), LYT-04 40℃ / 75%RH

[0299]

[0300]

[0301] 1 The sum of related substances> 0.05%

[0302] ND = Not Detected

[0303] LOQ = limit of quantitation

[0304] in conclusion

[0305] The results of Example 2 show that LTX-315 is stable in aqueous solution in a sealed container at pH 5 to 7. At pH 8, an increase in related substances is seen, which is also reflected in a decrease in concentration and purity.

[0306] Example 3: Aqueous formulations for the medical products of the present invention

[0307] The aqueous preparation in Example 3 can be prepared using a conventional preparation method, comprising the following steps:

[0308] 1. Weigh the buffer components and prepare the required amount of aqueous buffer and set aside.

[0309] 2. Add approximately half of the buffer to a vessel equipped with a stirring device and optional heating device, such as a flask and a magnetic stirrer, and stir.

[0310] 3. Add excipients to the buffer while stirring.

[0311] 4. Finally, add the remaining buffer, stir vigorously, and continue stirring until the excipient is completely dissolved.

[0312] The peptide can be added in step 2 or step 4. In step 2, the buffer can be heated to about 70°C to facilitate dissolution of certain excipients. If the buffer is heated, the remainder of the buffer in step 4 is preferably at room temperature, and the peptide is preferably added in step 4. Sterile formulations can be prepared by filtering the formulation through a 0.22 μm sterile filter into a container and sealing the container.

[0313] 3a. 12 mg / mL LTX-315 aqueous formulation containing viscosity enhancer HPMC (low molecular weight).

[0314] HPMC was obtained from Safic Alcan, Paris La Dèfense Cedex, France.

[0315] Table 23:

[0316] Element Weight (g) %w / w LTX-315 acetate 1.2 1.0 Low molecular weight HPMC 0.5 0.5 Acetic acid / acetate buffer 0.2 M, pH 5.8 Ad 100 98

[0317] The resulting aqueous preparation had a pH of 5.8, a viscosity of 45 cP, and an osmotic pressure of 380 mmol / L.

[0318] 3b. 6 mg / mL aqueous formulation of LTX-315, containing the viscosity enhancer HPMC (high molecular weight).

[0319] HPMC was obtained from Safic Alcan, Paris La Dèfense Cedex, France.

[0320] Table 24:

[0321] Element Weight (g) %w / w LTX-315 acetate 0.6 0.6 High molecular weight HPMC 0.5 0.5 Acetic acid / acetate buffer 0.2 M, pH 5.8 Ad 100 99

[0322] The resulting aqueous preparation had a pH of 5.8, a viscosity of 150 cP, and an osmotic pressure of 380 mmol / L.

[0323] 3c. 10 mg / mL LTX-315 aqueous formulation containing chitosan (low molecular weight), a viscosity enhancer.

[0324] Chitosan was obtained from Chitinor AS, Norway.

[0325] Table 25:

[0326] Element Weight (g) %w / w LTX-315 acetate 1.0 1.0 Low molecular weight chitosan 1.0 1.0 Acetic acid / acetate buffer 0.2 M, pH 5.2 Ad 100 98

[0327] The pH of the obtained aqueous preparation was 5.6, the viscosity was 60 cP, and the osmotic pressure was 350 mmol / L.

[0328] 3d. 16 mg / mL LTX-315 aqueous formulation containing chitosan (medium molecular weight), a viscosity enhancer.

[0329] Chitosan was obtained from Chitinor AS, Norway.

[0330] Table 26

[0331] Element Weight (g) %w / w LTX-315 acetate 1.6 1.6 Medium molecular weight chitosan 1.0 1.0 Acetic acid / acetate buffer 0.2 M, pH 5.2 Ad 100 97

[0332] The pH of the obtained aqueous preparation was 5.6, the viscosity was 120 cP, and the osmotic pressure was 350 mmol / L.

[0333] 3e. 16 mg / mL LTX-302 aqueous formulation containing chitosan (medium molecular weight) as a viscosity enhancer and benzene as a preservative Methanol

[0334] Table 27:

[0335] Element Weight (g) %w / w LTX-302 acetate 1.6 1.6 Medium molecular weight chitosan 0.5 0.5 Benzyl alcohol 1.0 1.0 Phosphate buffer 0.2 M, pH 6.0 Ad 100 97

[0336] 3f. 8mg / mL LTX-302 aqueous formulation, containing the viscosity enhancer chitosan (low molecular weight) and the chelating agent EDTA

[0337] Table 28:

[0338] Element Weight (g) %w / w LTX-302 acetate 0.8 0.8 Low molecular weight chitosan 0.5 0.5 Disodium EDTA 0.1 0.1 Phosphate buffer 0.2 M, pH 6.0 Ad 100 99

[0339] 3g of 10mg / mL LTX-320 aqueous formulation containing the antioxidant sodium bisulfite

[0340] Table 29:

[0341] Element Weight (g) %w / w LTX-320 trifluoroacetate 1.0 1.0 Sodium bisulfite 0.1 0.1 Citrate buffer 0.2 M, pH 6.0 Ad 100 99

[0342] 3h. 10 mg / mL LTX-313 aqueous formulation, containing stabilizer PEG 200

[0343] Table 30:

[0344] Element Weight (g) %w / w LTX-313 acetate 1.0 0.9 PEG 200 5.0 4.7 Citrate buffer 0.2 M, pH 6.0 Ad 100 94

[0345] Example 4: Stability Investigation

[0346] Vitas Analytical Services in Norway conducted a stability study of LTX-315 in different formulations stored at 23°C ± 2°C and 2°C to 8°C. The prepared sterile, filtered formulations were stored in Ph. Eur. Type 1 6R vials (Schott AG), stoppered with Ph. Eur. Type 1 lyophilized rubber stoppers V9172 FM257 / 2ISAF1 (Datwyler PharmaPackaging), and capped with flip-top aluminum caps (West Pharmaceutical Services). The formulations were stored at 2°C to 8°C and 23°C ± 2°C. The parameters assay, purity, related impurities, visual appearance, and pH were tested for up to four months of storage.

[0347] The formulations are summarized below. Formulations 3 to 6 contained 10 mg / mL LTX-315 (neat peptide):

[0348]

[0349] Material

[0350] Preparation of buffer:

[0351] Acetate buffer 0.1M pH 5.2:

[0352]

[0353]

[0354] qs: appropriate amount

[0355] Phosphate buffer 0.1 M pH 6.0

[0356] composition Disodium hydrogen phosphate dihydrate 8.9g Potassium dihydrogen phosphate 6.8g NaOH / HCl qs WFI Ad 1000ml

[0357] The buffer was filtered through a 0.22 μm sterile filter into a flask rinsed with WFI that had been filtered through a 0.22 μm sterile filter or into an autoclaved flask.

[0358] Preparation of the preparation:

[0359] LTX-315 is supplied in sealed vials containing 20 mg of sterile, lyophilized peptide as its acetate salt.

[0360] Preparation 1

[0361]

[0362] Preparation 2

[0363]

[0364] For Formulations 1 and 2, 2 mL of the corresponding sterile buffer was injected into the vial containing LTX-315 and LTX-315 was dissolved in the buffer. Two vials were prepared for stability studies, one vial was stored at 2°C to 8°C, and the other vial was stored at 23°C ± 2°C.

[0365] Preparation 3

[0366]

[0367]

[0368] Preparation 4

[0369]

[0370] Preparation 5

[0371]

[0372] Chitosan: low molecular weight, 330 kDa, DAA 86% (Manufacturer: Chitinor, Norway).

[0373] Preparation 6

[0374]

[0375] HPMC: approximately 86 kDA (Manufacturer: Sigma Aldrich)

[0376] For Formulations 3 to 6, excipient formulations in acetate buffer were prepared as described in Example 3 and filtered through a 0.22 μm sterile filter. 2 mL of the corresponding sterile formulation was injected into a vial containing LTX-315, and LTX-315 was dissolved in the formulation. Two vials were prepared for stability studies, one stored at 2°C to 8°C and the other at 23°C ± 2°C.

[0377] Assay, purity and related impurities

[0378] method

[0379] 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 x 3.0 mm ID column at 15°C and 0.612 mL / min with an ACN / H2O mobile phase gradient for a 38-minute run time. The injection volume was 2.0 μL, and the injection temperature was 5°C.

[0380] Purity is calculated by subtracting the measured impurities (total organic impurities) from 100, and the relevant impurities are reported by comparing the diluent chromatogram with the sample chromatogram. Known peaks present in the diluent chromatogram with the same intensity for both solutions are ignored. This method has been validated and is suitable for its intended use.

[0381] The pH was determined according to European Pharmacopoeia 2.2.3.

[0382] Appearance (clarity and color) was determined by visual inspection.

[0383] Results and Conclusions

[0384] Under each storage condition, the pH and appearance of all formulations remained essentially unchanged after 4 months.

[0385] like Figure 1 and Figure 2 As shown, no significant changes in the assay were observed for any of the formulations after 4 months of storage at either 2°C to 8°C or 23°C ± 2°C. The minor changes observed were due to method variability.

[0386] At the initial time point (T0), no impurities were observed in any formulation, and thus the purity of all formulations was observed to be 100%. No significant changes were observed in any formulation during storage for up to 4 months at 2°C to 8°C, as shown in Table 1. Figure 3 shown.

[0387] For the preparations stored at 23°C ± 2°C, it was observed that the purity of preparation 4 decreased to 98.4% after 1 month (T1) and further decreased to 97.6% after 4 months (T4). Figure 4 This decrease in purity corresponds to an increase of 1.6% and 2.4% in total related impurities after 1 month and 4 months, respectively.

[0388] When stored at 23°C ± 2°C for 4 months, the purity of Formulation 3 was observed to drop to 98.6%, which corresponded to an increase in the total related impurities to 1.4%.

[0389] in conclusion

[0390] Based on the above, LTX-315 was stable in various aqueous solutions at temperatures between 2°C and 8°C in sealed containers, as there was essentially no change in any of the parameters tested. At 23°C ± 2°C, the stability results showed some instability in Formulations 3 and 4, but these were not considered "significant changes" according to ICH Q1A (Stability Testing of New Drug Substances and Products).

[0391] Implementation Plan

[0392] 1. A medicinal product in a sealed container comprising an aqueous preparation of a peptide or a pharmaceutically acceptable salt thereof, wherein the peptide satisfies:

[0393] a) consists of 9 linearly arranged amino acids;

[0394] b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and

[0395] c) Of these four lipophilic amino acids, three are tryptophans and one is a non-genetically encoded amino acid.

[0396] 2. The medical product according to embodiment 1, wherein the cationic amino acid in the peptide is lysine or arginine.

[0397] 3. The medical product according to any one of embodiments 1 to 2, wherein the non-genetically encoded amino acid in the peptide is diphenylalanine or biphenylalanine.

[0398] 4. The medical product according to any one of embodiments 1 to 3, wherein the peptide has an amino acid sequence selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8 and 9.

[0399] 5. The medical product according to any one of embodiments 1 to 4, wherein the peptide has an amino acid sequence selected from SEQ ID NO: 1, 2 and 7.

[0400] 6. The medical product according to any one of embodiments 1 to 5, wherein the peptide has an amino acid sequence selected from SEQ ID NOs: 10 to 42.

[0401] 7. The medical product according to any one of embodiments 1 to 6, wherein the peptide is selected from LTX-302, LTX-313, LTX-315, LTX-320 and LTX-329.

[0402] 8. The medical product according to any one of embodiments 1 to 7, wherein the peptide is in the form of a pharmaceutically acceptable salt.

[0403] 9. The medical product according to any one of embodiments 1 to 8, wherein the peptide is in the form of acetate.

[0404] 10. The medical product according to any one of embodiments 1 to 9, wherein the peptide is LTX-315 having the amino acid sequence of SEQ ID NO: 23.

[0405] 11. The medical product according to 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.

[0406] 12. The medical product according to 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 acetate salt.

[0407] 13. The medical product according to any one of embodiments 1 to 12, wherein the concentration of the peptide in the aqueous formulation is 1 mg / mL to 30 mg / mL, preferably 4 mg / mL to 20 mg / mL, or more preferably 6 mg / mL to 16 mg / mL, for example 10 mg / mL.

[0408] 14. The medical product according to embodiment 13, wherein the amount of the peptide is the net amount of peptide.

[0409] 15. The medical product according to any one of embodiments 1 to 14, wherein the aqueous formulation comprises water for injection.

[0410] 16. The medical product according to any one of embodiments 1 to 15, wherein the aqueous formulation consists of the peptide and water for injection.

[0411] 17. The medical product according to any one of embodiments 1 to 15, wherein the aqueous preparation comprises an aqueous buffer, for example, the buffer is selected from: acetic acid buffer, acetate buffer, aspartic acid buffer, sodium benzoate buffer, benzoic acid buffer, carbonate buffer, citric acid buffer, glucono-δ-lactone buffer, glycine buffer, glycine HCl buffer, histidine buffer, histidine HCl buffer, hydrobromic acid buffer, phosphate buffer (e.g., PBS), sodium succinate buffer, disodium succinate buffer, succinic acid buffer, sulfuric acid buffer, tartaric acid buffer and sodium tartrate buffer, preferably selected from: acetic acid / acetate buffer, citric acid buffer and phosphate buffer.

[0412] 18. The medical product according to embodiment 17, wherein the aqueous buffer is a mixture of several different buffers.

[0413] 19. The medical product according to any one of embodiments 1 to 15 and 17 to 18, wherein the aqueous formulation consists of a peptide and an aqueous buffer.

[0414] 20. The medicinal product according to any one of embodiments 1 to 15 and 17 to 18, wherein the aqueous formulation comprises one or several salts.

[0415] 21. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20, wherein the aqueous formulation further comprises at least one preservative.

[0416] 22. The medical product according to embodiment 21, wherein the preservative is selected from the group consisting of benzalkonium chloride, benzyl alcohol, chlorobutanol, m-cresol, methylparaben, phenol, potassium sorbate, propylparaben and thimerosal.

[0417] 23. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20 to 22, wherein the aqueous formulation further comprises at least one chelating agent.

[0418] 24. The medical product according to embodiment 23, wherein the chelating agent is selected from the group consisting of calcium disodium EDTA, sodium calcium veselamide, carboxydoline, disodium EDTA, DTPA, and sodium EDTA.

[0419] 25. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20 to 24, wherein the aqueous formulation further comprises at least one antioxidant.

[0420] 26. The medical product according to embodiment 25, wherein the antioxidant is selected from the group consisting of ascorbic acid, ascorbyl palmitate, citric acid, isoascorbic acid, methionine, monothioglycerol, potassium metabisulfite, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium thiosulfate, and thiourea.

[0421] 27. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20 to 26, wherein the aqueous formulation further comprises at least one osmotic pressure regulator.

[0422] 28. The medical product according to embodiment 27, wherein the osmotic pressure regulator is selected from the group consisting of glucose, glycerol, lactose, mannitol, potassium chloride, sodium chloride and sorbitol, preferably sodium chloride.

[0423] 29. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20 to 28, wherein the aqueous formulation further comprises at least one stabilizer.

[0424] 30. The medical product according to 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.

[0425] 31. The medical product according to any one of embodiments 1 to 15, 17 to 18 and 20 to 28, wherein the aqueous formulation further comprises at least one viscosity enhancing agent.

[0426] 32. The medical product according to embodiment 29, wherein the viscosity enhancer is selected from the group consisting of methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose, carboxymethyl cellulose calcium, carboxymethyl cellulose sodium, chitosan, gelatin, hyaluronic acid, maltodextrin, pectin, poloxamer 407, PEG, polyethylene oxide, povidone (PVP), starches such as corn starch and tapioca starch, tragacanth gum, and β-cyclodextrin.

[0427] 33. The medical product according to embodiment 32, wherein the viscosity enhancer is chitosan or a chitosan derivative.

[0428] 34. The medical product according to any one of embodiments 1 to 33, wherein the pH of the aqueous formulation is 5.0 to 7.4, preferably 5.0 to 7.0, more preferably 5.0 to 6.0, for example 6.5.

[0429] 35. The medical product according to any one of embodiments 1 to 34, wherein the viscosity of the aqueous formulation is 1 cP, 1.5 cP, 2 cP, 2.5 cP or 3 cP to 300 cP, such as 2.5 cP to 250 cP, such as 5 cP to 100 cP or 10 cP to 200 cP and 20 cP to 240 cP, preferably 40 cP to 180 cP, such as 45 cP to 150 cP or 30 cP to 100 cP, more preferably 1.5 cP to 10 cP, such as 2 cP, 3 cP, 4 cP, 5 cP, 6 cP, 7 cP, 8 cP, 9 cP or 10 cP, or 10 cP to 15 cP or 15 cP to 20 cP or 20 cP to 35 cP or 35 cP to 50 cP or 50 cP to 100 cP.

[0430] 36. A medicinal product according to any one of embodiments 1 to 35, wherein the total amount of excipients present in the aqueous formulation does not exceed 20% w / w, preferably does not exceed 5% w / w, more preferably does not exceed 4% w / w, for example does not exceed 3% w / w or does not exceed 2% w / w.

[0431] 37. The medicinal product of any one of embodiments 1 to 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, less than 2% w / w or less than 1% w / w.

[0432] 38. The medical product according to any one of embodiments 1 to 37, wherein the aqueous formulation is a sterile formulation.

[0433] 39. The medical product according to any one of embodiments 1 to 38, wherein the aqueous preparation is a ready-to-use injection preparation, preferably a ready-to-use injection solution of the peptide.

[0434] 40. The medical product according to any one of embodiments 1 to 39, wherein the sealed container is sealed with a manufactured seal.

[0435] 41. The medical product according to any one of embodiments 1 to 40, wherein the sealed container is selected from the group consisting of: a prefilled syringe, a syringe cartridge, a vial, and an ampoule.

[0436] 42. The medical product according to any one of embodiments 1 to 41, wherein the usable volume of the sealed container is 0.1 mL to 20 mL, such as 0.2 mL to 10 mL, such as 0.3 mL to 8 mL, and preferably 0.5 mL to 5 mL.

[0437] 43. The medical product according to any one of embodiments 1 to 42, wherein the volume of the aqueous formulation contained in the sealed container is 0.1 mL to 20 mL, such as 0.2 mL to 10 mL, preferably 0.2 mL to 5.0 mL or 10 mL.

[0438] 44. The medical product according to any one of embodiments 1 to 43, wherein the sealed container contains a single dose.

[0439] 45. The medical product according to any one of embodiments 1 to 43, wherein the sealed container contains multiple doses.

[0440] 46. ​​A method of manufacturing a medical product according to any one of embodiments 1 to 45, wherein the aqueous formulation is introduced into a container and the filled container is mechanically sealed.

[0441] 47. A method of treating a tumor in a subject, the method comprising obtaining the medical product of any one of embodiments 1 to 45 and administering the aqueous formulation contained therein to the subject.

[0442] 48. The method of embodiment 47, wherein the aqueous formulation is administered into a tumor of the subject.

[0443] 49. A method according to any one of embodiments 47 to 48, wherein the tumor is selected from: carcinoma (e.g., hepatocellular carcinoma), adenocarcinoma, sarcoma, melanoma, lymphoma, leukemia, germ cell tumor, blastoma and glioblastoma, and the tumor is associated with the following cancers: nervous system cancer, brain cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer and head and neck cancer.

[0444] 50. The method of embodiment 49, wherein the tumor is selected from the group consisting of: melanoma, carcinoma, sarcoma, and lymphoma, and the tumor is associated with a cancer selected from the group consisting of: skin cancer, breast cancer, and head and neck cancer.

Claims

1. A medicinal product in a sealed container comprising an aqueous preparation of a peptide or a pharmaceutically acceptable salt thereof, wherein the peptide satisfies: a) consists of 9 linearly arranged amino acids; b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and c) Of these four lipophilic amino acids, three are tryptophans and one is a non-genetically encoded amino acid.

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

3. The medical product according to 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, and is preferably in the form of a pharmaceutically acceptable salt, preferably an acetate salt.

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

5. The medical product according to any one of claims 1 to 4, wherein The concentration of the peptide in the aqueous formulation is 1 mg / mL to 30 mg / mL, preferably 4 mg / mL to 20 mg / mL.

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

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

8. The medical product according to 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, osmotic pressure regulators, stabilizers, antioxidants, viscosity enhancers, and chelating agents.

9. The medical product according to any one of claims 1 to 8, wherein The aqueous formulation also includes at least one viscosity enhancing agent.

10. The medical product according to claim 9, wherein The viscosity enhancer is chitosan or a chitosan derivative.

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

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

4.

13. The medical product according to any one of claims 1 to 12, wherein The total amount of excipients present in the aqueous formulation does not exceed 20% w / w, preferably does not exceed 5% w / w.

14. The medical product according to any one of claims 1 to 6 and 8 to 13, 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.

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

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

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

18. The medical product according to any one of claims 1 to 17, wherein The volume of the aqueous formulation contained in the sealed container is 0.1 mL to 20 mL, preferably 0.2 mL to 10 mL or 0.2 mL to 5 mL.

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

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

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

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

24. The method according to 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 the tumor is associated with a cancer selected from the group consisting of cancer of the nervous system, brain, breast, colon, kidney, liver, lung, ovarian, pancreatic, prostate, and head and neck.

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

26. Use of a peptide or a pharmaceutically acceptable salt thereof for the preparation of a medicinal product in the form of a sealed container comprising an aqueous formulation of the peptide or a pharmaceutically acceptable salt thereof for treating a tumor in a subject, wherein the peptide satisfies: a) consists of 9 linearly arranged amino acids; b) of these 9 amino acids, 5 are cationic amino acids and 4 are lipophilic amino acids; and c) Of these four lipophilic amino acids, three are tryptophans and one is a non-genetically encoded amino acid. in, The aqueous formulation contained in the sealed container is administered to the subject.

Images