Compositions and Uses

Abstract

The disclosure herein relates to compositions, kits, and their use for treating subjects, e.g., patients, with tumors by administering such compositions to the subject.

Description

[Technical Field]

[0001] The disclosure herein relates to compositions, kits, and their use for treating subjects, e.g., patients, with tumors by administering such compositions to the subject. [Background technology]

[0002] Oncolytic peptides are highly effective anticancer 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, leading to their lysis. This disruption of the organelle membranes leads to the release of agents with potent immunostimulatory functions, commonly known as damage-associated molecular pattern molecules (DAMPs), including ATP, cytochrome C, mitochondrial CpG DNA sequences, mitochondrial formyl peptides, cathepsins (from lysosomes), and HMGB1 (from the nucleus). Consequently, these oncolytic peptides stimulate the immune system to elicit an immune response, thereby generating long-term antitumor immunity. The immune stimulation that occurs when oncolytic peptides are injected into tumors and their direct lytic effect on the tumor releases DAMPs and tumor antigens, promoting antigen presentation and thus providing a kind of in situ "vaccination" against the tumor.

[0003] Although oncolytic peptides are promising tools in the immunotherapy of cancer, there is still room for improvement in their use in cancer therapy. Summary of the Invention [Means for solving the problem]

[0004] In a first aspect, the present disclosure relates to a combination of composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative for use in treating a subject having a tumor, or to composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative for use in treating a subject having a tumor.

[0005] In some embodiments, compositions (a) and (b) are used simultaneously for treatment. In some other embodiments, compositions (a) and (b) are used sequentially for treatment. In yet some other embodiments, compositions (a) and (b) are combined into a single composition for treatment, i.e., a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative.

[0006] Thus, in some embodiments, a first aspect of the present disclosure relates to a combination of composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative for simultaneous, sequential, or combined use in treating a subject having a tumor, or to composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative for simultaneous, sequential, or combined use in treating a subject having a tumor, wherein, for example, compositions (a) and (b) are administered to said subject simultaneously, sequentially, or as a combined composition.

[0007] In an alternative first aspect, the present disclosure relates to a composition (a) comprising an oncolytic peptide for use in treating a subject having a tumor by simultaneous, sequential, or combined administration with a composition (b) comprising chitosan and / or a chitosan derivative.

[0008] In a further alternative first aspect, the present disclosure relates to a method of treating a subject having a tumor, the method comprising administering to said subject a composition (a) comprising an oncolytic peptide and a composition (b) comprising chitosan and / or a chitosan derivative simultaneously, sequentially, or in combination.

[0009] In a further alternative first aspect, the present disclosure relates to the use of composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative for treating a subject having a tumor. In some embodiments, compositions (a) and (b) are used simultaneously, sequentially, or as a combined composition to treat the subject, e.g., compositions (a) and (b) are administered to the subject simultaneously, sequentially, or as a combined composition.

[0010] In some embodiments, the composition comprises a physiologically acceptable carrier. In some embodiments, composition (a) is administered into the tumor and composition (b) is administered into the tumor and / or into the tissue immediately surrounding the tumor.

[0011] In some embodiments, the composition is a pharmaceutical composition.

[0012] In some embodiments, the composition comprising an oncolytic peptide and the composition comprising chitosan and / or a chitosan derivative are combined into one composition.

[0013] Thus, in some embodiments, the present disclosure relates to a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative for use in treating a subject having a tumor, in some embodiments, the composition is administered to the subject, e.g., administered intratumorally to the subject.

[0014] Alternatively, the present disclosure relates to a method of treating a subject having a tumor, the method comprising administering to the subject a composition comprising: i) an oncolytic peptide; and ii) chitosan and / or a chitosan derivative. In some embodiments, the composition is administered intratumorally.

[0015] In another alternative, the present disclosure relates to the use of i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative for the manufacture of a medicament for use in treating a subject having a tumor, in some embodiments, the medicament being administered to the subject, e.g., intratumorally in the subject.

[0016] In yet another alternative, the present disclosure relates to a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative for treating a subject having a tumor, in some embodiments, the composition is administered to the subject, e.g., administered intratumorally to the subject.

[0017] In yet another alternative, the present disclosure relates to a method for treating a tumor in a subject by administering the drug to the subject, the drug comprising: i) an oncolytic peptide; and ii) chitosan and / or a chitosan derivative. In some embodiments, the drug is administered intratumorally.

[0018] In some embodiments, the composition comprises a physiologically acceptable carrier. In some embodiments, the composition is a pharmaceutical composition.

[0019] In another aspect, the present disclosure relates to a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative.

[0020] In some embodiments, the composition comprises a physiologically acceptable carrier.

[0021] In some embodiments, the composition is a pharmaceutical composition.

[0022] In yet another aspect, the present disclosure relates to a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative thereof for use as a medicament.

[0023] In some embodiments, the composition comprises a physiologically acceptable carrier.

[0024] In some embodiments, the composition is a pharmaceutical composition.

[0025] In yet another aspect, the present disclosure relates to a method for preparing a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative.

[0026] In some embodiments, the composition comprises a physiologically acceptable carrier and / or is sealed in a container.

[0027] In some embodiments, the composition is a pharmaceutical composition.

[0028] In yet another aspect, the present disclosure relates to a kit comprising composition (a) comprising an oncolytic peptide and composition (b) comprising chitosan and / or a chitosan derivative, or comprising parts / components for preparing compositions (a) and (b).

[0029] In yet another aspect, the present disclosure relates to a kit comprising parts / components for preparing a composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative. [Brief explanation of the drawings]

[0030] [Figure 1]Figure 1 shows the results of Example 3, i.e., the therapeutic effect of a composition comprising LTX-401 and chitosan in tumor-bearing mice compared with a composition comprising LTX-401 alone, LTX-401 and HPMC, or a control. Therapeutic efficacy is shown as the % of mice with tumor volumes less than 1 ml / day from the start of treatment. [Figure 2] Figure 2 shows the results of Example 4, i.e., the therapeutic effect of a composition comprising LTX-401 and chitosan in tumor-bearing mice compared with a composition comprising LTX-401 alone, chitosan alone, or a control. Therapeutic efficacy is expressed as % survival rate / days after tumor challenge. DETAILED DESCRIPTION OF THE INVENTION

[0031] A "subject" is an animal, e.g., a mouse or a human, preferably a human. A subject may be a patient, i.e., a human with cancer who is in need of treatment for the cancer or who is in need of treatment to prevent the recurrence of the cancer. As used herein, the terms "subject" and "individual" are used interchangeably.

[0032] As used herein, "cancer" refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells in the body, resulting in a tumor. A "tumor" or "cancerous tissue" or "(cancerous) lesion" includes primary tumors and metastases. Tumors include solid and non-solid tumors.

[0033] As used herein, "treatment" is a therapeutic treatment.

[0034] A "therapeutic treatment" is a treatment administered to a subject who exhibits symptoms or signs of cancer, wherein the treatment is administered to the subject for the purpose of reducing or eliminating those signs or symptoms, or for the purpose of slowing or halting the progression of the disease, for example, stopping or slowing the growth or increase in malignancy or the spread of a tumor.

[0035] The term "oncolytic peptide" as used herein refers to oncolytic positively charged amphiphilic amino acid derivatives, peptides, or peptidomimetics, i.e., positively charged amphiphilic amino acid derivatives, peptides, or peptidomimetics that can dissolve tumor cell membranes.Because of their amphiphilic nature, they have one or more hydrophilic groups, i.e., cationic groups, and one or more hydrophobic groups.Therefore, these molecules a) are attracted to the negative charge of tumor cell membranes, and b) can interact with the hydrophobic tails of phospholipids contained in tumor cell membranes through their hydrophobic group(s), which are composed of fatty acid chains.

[0036] Oncolytic peptides As mentioned above, the oncolytic peptide for use in the present disclosure refers to oncolytic positively charged amphipathic amino acid derivative, peptide or peptidomimetic.Oncolytic peptides are described in patent publications WO2007 / 107748(A2), WO2010 / 060497(A1), WO2011 / 051692(A1), WO2016 / 091487(A1) and WO2016 / 091490(A1), the contents of which are incorporated herein in their entirety.

[0037] 9-mer peptide In some embodiments, an oncolytic peptide for use in the present disclosure is a 9-mer peptide or a pharmaceutically acceptable salt thereof consisting of nine amino acids in a linear arrangement, five of which are cationic and four of which are lipophilic, and three of which are tryptophan and one of which is a non-genetically encoded amino acid.

[0038] In some embodiments, in the above-described 9-mer peptides, the lipophilic and cationic amino acids are arranged such that no more than two amino acids of either type are adjacent to one another and / or such that the peptide contains two pairs of adjacent cationic amino acids and one or two pairs of adjacent lipophilic amino acids.

[0039] The term "amino acid" as used in defining the 9-mer peptide includes alpha, beta, and gamma amino acids as well as N-substituted glycines.

[0040] "Non-genetically encoded amino acids" are amino acids that differ not only stereospecifically but also structurally from the genetically encoded L-amino acids. Thus, as disclosed herein, D-amino acids, although not strictly genetically encoded, are not considered non-genetically encoded amino acids.

[0041] The cationic amino acids may be the same or different. In some embodiments, the cationic amino acid is lysine and / or arginine. In some other embodiments, the cationic amino acid is histidine and / or a non-genetically encoded amino acid that has a positive charge at pH 7.0. Lysine and arginine are preferred cationic amino acids.

[0042] Non-genetically encoded cationic amino acids include derivatives of lysine, arginine, and histidine, such as homolysine, ornithine, diaminobutyric acid, diaminopimelic acid, diaminopropionic acid, homoarginine, trimethylysine, trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-carbamimidoylpiperidine-4-carboxylic acid, and 4-guanidinophenylalanine.

[0043] The non-genetically encoded lipophilic amino acids may be the same or different.

[0044] In some embodiments, the non-genetically encoded lipophilic amino acid has a lipophilic side chain having at least 7, preferably at least 8 or 9, more preferably at least 10 non-hydrogen atoms, preferably carbon atoms. In some embodiments, the lipophilic side chain contains 30 or fewer non-hydrogen atoms, more preferably 25 or fewer non-hydrogen atoms. In some embodiments, the lipophilic side chain comprises at least one, preferably two, cyclic groups, which may be fused or connected.

[0045] In some embodiments, the lipophilic side chain contains a heteroatom such as O, N, or S, although typically no more than one heteroatom will be present, which is preferably nitrogen. The lipophilic side chain will preferably have no more than two polar groups, more preferably zero or one, and most preferably none.

[0046] Preferred non-genetically encoded lipophilic 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)- Particularly preferred non-genetically encoded lipophilic 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 non-genetically encoded lipophilic amino acids.

[0047] In some embodiments, some or all of the amino acids present in the aforementioned peptides may be in the D-form.

[0048] In a preferred embodiment, the 9-mer peptide is a compound of formula (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. 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 its own 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)

[0049] As discussed above, the peptide contains one non-genetically encoded lipophilic amino acid. When this amino acid is designated as 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)

[0050] Particularly preferred are the 9-mer peptides of formula (I) and (II), of which the peptide of formula (I'') is particularly preferred.

[0051] Most preferred are the 9-mer peptides listed in the table below.

[0052] [Table 1]

[0053] 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

[0054] Preferred 9-mer 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.

[0055] For use in the present invention, all of the 9-mer peptides described herein may be in the form of pharmaceutically acceptable salts.Peptides preferably have modified C-terminus, 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.

[0056] Particularly preferred is the 9-mer peptide known as LTX-315, especially in the form of a pharmaceutically acceptable salt, preferably the acetate salt, such as LTX-315 having the amino acid sequence of SEQ ID NO:23, or a pharmaceutically acceptable salt thereof, preferably the acetate salt thereof.

[0057] The 9-mer peptides and pharmaceutically acceptable salts thereof for use in the present disclosure may be synthesized in any convenient manner by peptide synthesis methods well known in the art, preferably by carrying out synthesis on a solid support. Synthetic methods are described, for example, in WO 2010 / 060497 (A1), WO 2016 / 091487 (A1), and WO 2016 / 091490 (A1).

[0058] Compounds containing disubstituted β-amino acids In some embodiments, an oncolytic peptide for use in the present disclosure is a peptide, peptidomimetic, or amino acid derivative having a net positive charge of at least +2 and including disubstituted β-amino acids, wherein each of the substituents, which may be the same or different, of the β-amino acids contains at least 7 non-hydrogen atoms, is lipophilic, and has at least one cyclic group, wherein one or more cyclic groups in a substituent may be linked or fused with one or more cyclic groups in the other substituent, and wherein the combined total number of non-hydrogen atoms for the two substituents when the cyclic groups are fused in this manner is at least 12. The two substituents in the β-amino acid are preferably the same.

[0059] β-amino acids have an amino group attached to the β-carbon atom. Genetically encoded amino acids are α-amino acids, where the amino group is attached to the α-carbon atom. This arrangement lengthens the backbone of a peptide containing one or more β-amino acids by one atom per β-amino acid. In this arrangement, the α- and / or β-carbon atoms can be substituted. The α- or β-carbon atoms can be disubstituted, where di-substitution of the α-carbon atom results in a β- 2,2 When the β carbon atom is disubstituted, an amino acid is obtained.3,3 Amino acids are produced. One substituent at each of the α or β carbon atoms is 2,3 Produces amino acids. 2,2 and β 3,3 Disubstituted amino acids are preferred for use in accordance with the present invention, 2,2 Disubstituted amino acids are particularly preferred.

[0060] The β-amino acid is substituted with two groups containing at least seven non-hydrogen atoms. Preferably, one, more preferably both, of the substituents contains at least eight, more preferably at least ten non-hydrogen atoms. These groups are lipophilic in nature, and they may be different but are preferably the same. Each contains at least one cyclic group, typically a six-membered ring, which may be aliphatic or aromatic, preferably aromatic, and may be substituted. The substituents may contain heteroatoms, such as oxygen, nitrogen, sulfur, or halogens, particularly fluorine or chlorine. Preferred substituents include C1-C4 alkyl (especially t-butyl), methoxy, fluoro, and fluoromethyl groups. The cyclic groups may be homocyclic or heterocyclic, preferably a homocyclic ring of carbon atoms. Preferred lipophilic substituents include two or three cyclic groups, preferably two cyclic groups, which may be connected or fused, preferably fused. A particularly preferred substituent includes a naphthalene group.

[0061] A more preferred group of lipophilic substituents contains a single substituted or unsubstituted cyclic group, preferably a phenyl or cyclohexyl group.

[0062] Typically, the cyclic group(s) are spaced from the peptide backbone (i.e., the α or β carbon atom of a β amino acid) by a chain of 1 to 4, preferably 1 to 3, atoms. These linking atoms may include nitrogen and / or oxygen, but are typically carbon atoms, and preferably the linking atoms are unsubstituted. Of course, these spacers are part of the substituents defined herein.

[0063] Each substituted moiety of a disubstituted β-amino acid will typically contain between 7 and 20 non-hydrogen atoms, preferably between 7 and 13, more preferably between 8 and 12, and most preferably between 9 and 11 non-hydrogen atoms.

[0064] Compounds containing disubstituted β-amino acids are preferably peptides or peptidomimetics of 1 or 2-12 amino acids or equivalent subunits in length. Unless otherwise clear from the context, reference herein to an "amino acid" includes the equivalent subunits in a peptidomimetic. Preferred compounds contain 3-12 amino acids in length, more preferably 5-12 amino acids in length. In some embodiments, compounds containing disubstituted β-amino acids consist of such disubstituted β-amino acids, i.e., they are amino acid derivatives (i.e., modified amino acids).

[0065] Compounds containing disubstituted β-amino acids preferably include a modified C-terminus, and the C-terminus modification typically results in charge reversal, i.e., by removing the negative charge of the carboxyl group and adding a positive charge, for example, by the presence of an amino group. This modification alone, assuming that the N-terminus is not modified, gives the entire molecule a net charge of +2. Whether the C-terminus is modified to provide charge reversal or simply to remove the negative charge of the carboxyl group, in some embodiments where the compound is a peptide or peptidomimetic, such compounds also contain one or more cationic amino acids. Thus, the overall charge of the compound may be +3 or +4 or higher.

[0066] Suitable C-terminal groups are preferably cationic in nature and typically have a maximum size of 15 non-hydrogen atoms. The C-terminus is preferably amidated, and the amide group may be further substituted to form N-alkyl or N,N-dialkylamides. Primary and secondary amide groups are preferred. Suitable groups for substituting the amide group include aminoalkyls, such as aminoethyl or dimethylaminoethyl. The nitrogen atom of the amide group may form part of a cyclic group, such as pyrazolidine, piperidine, imidazolidine, and piperazine, with piperazine being preferred, which cyclic groups may themselves be substituted, for example, by alkyl or aminoalkyl groups.

[0067] In some embodiments, the disubstituted β-amino acid-containing compound is a peptide that includes one or more cationic amino acids. Lysine, arginine, ornithine, and histidine are preferred, but any non-genetically encoded or modified amino acid that has a positive charge at pH 7.0 may be included.

[0068] 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.

[0069] In some embodiments, compounds containing disubstituted β-amino acids are dipeptides, which will typically include one cationic amino acid. Longer peptides will usually include additional cationic amino acids, so that a peptide of 4 or 5 amino acids may have 2 or 3 cationic amino acids, and a peptide of 6-9 amino acids may have 3-6 cationic amino acids.

[0070] In some embodiments, the compound comprising a disubstituted β-amino acid comprises a β-amino acid bonded to a C-terminal L-argininamide residue. 2,2 Dipeptides containing disubstituted amino acids and having this configuration are preferred.

[0071] Peptides with three or more amino acids will typically have one or more additional lipophilic amino acids, i.e., amino acids with lipophilic R groups. Typically, the lipophilic R group has at least one, preferably two, cyclic groups, which may be fused or connected. The lipophilic R group may contain heteroatoms, such as O, N, or S, but typically no more than one heteroatom is present, preferably nitrogen. The R group preferably has no more than two polar groups, more preferably zero or one, and most preferably none.

[0072] Tryptophan is a preferred lipophilic amino acid, and the peptide preferably contains 1 to 3 tryptophan residues. Further genetically encoded lipophilic amino acids that may be included are phenylalanine and tyrosine.

[0073] Lipophilic amino acids may be non-genetically encoded, including genetically encoded amino acids with modified R groups.

[0074] Peptide mimetics are typically characterized by retaining the polarity, three-dimensional size, and function (biological activity) of their peptide equivalents, but often with more stable bonds replacing the peptide bonds. "Stable" refers to greater resistance to enzymatic degradation by hydrolases. Amide bond substitutes generally preserve many of the properties of amide bonds, such as conformation, steric bulk, electrostatic characteristics, and hydrogen-bonding potential. Chapter 14 of "Drug Design and Development," Krogsgaard, Larsen, Liljefors, and Madsen (eds.), 1996, Horwood Academic Publishers, provides a general discussion of techniques for the design and synthesis of peptidomimetics. In the present disclosure, because oncolytic peptides / compounds containing disubstituted β-amino acids react with tumor cell membranes rather than with the specific active site of an enzyme, some of the problems described for accurately mimicking affinity and efficacy or substrate function are not relevant, and peptidomimetics can be easily prepared based on a given peptide structure or motif of the required functional groups. Suitable amide bond surrogates include the following groups:N-Alkylation (Schmidt, R. et al., International Journal of Peptide and Protein Research, Int. J. Peptide Protein Res., 1995, 46, 47), retroinverse amides (Chorev, M. and Goodman, M., Accounts of Chemical Research (Acc. Chem. Res.), 1993, 26, 266), thioamides (Sherman, D.B. and Spatola, A.F., J. Am. Chem. Soc., 1990, 112, 433), thioesters, phosphonates, ketomethylenes (Hoffman, R.V. and Kim, H.O., J. Org. Chem., 1995, 60, 5107), hydroxymethylenes, fluorovinyls (Allmendinger, T. et al., Tetrahydron Letters, 1995, 60, 5107), Lett., 1990, 31, 7297), vinyl, methyleneamino (Sasaki, Y. and Abe, J. Chemical and Pharmaceutical Bulletin (Chem. Pharm. Bull.). 1997 45, 13), methylenethio (Spatola, A. F., Methods Neurosci. 1993, 13, 19), alkanes (Lavielle, S. et al., Int. J. Peptide Protein Res. 1993, 42, 270), and sulfonamides (Luisi, G. et al., Tetrahedron Lett. 1993, 34, 2391).

[0075] In some embodiments, a compound containing a disubstituted β-amino acid contains only the disubstituted β-amino acid and one additional amino acid, i.e., it is a compound with two amino acids joined by an amide bond. Such molecules may be considered dipeptides due to the amide bond. However, the amide bond in these compounds is actually non-scissile due to the disubstituted β-amino acid, and therefore these compounds may be considered peptidomimetics. For the purposes of this disclosure, such compounds (and larger molecules with more amino acids) are considered peptides rather than peptidomimetics due to the presence of amide bonds. This allows for unambiguous naming without the need to verify whether or to what extent a given amide bond is cleavable. In other words, when all amino acids in a compound containing a disubstituted β-amino acid described herein are linked by amide bonds, the compound is considered a peptide, even if one or more of the amide bonds are not readily cleavable.

[0076] In some embodiments, the comprising a disubstituted β amino is a peptidomimetic, which will typically have distinguishable subunits of approximately the same size and function as amino acids. Peptidomimetics generally have groups equivalent to the R groups of amino acids, and the discussion herein about suitable R groups and N- and C-terminal modification groups applies mutatis mutandis to peptidomimetic compounds.

[0077] As discussed in the above-referenced texts, peptidomimetics, in addition to replacing amide bonds, may also involve replacing larger structural moieties with di- or tripeptidomimetic structures, in which case mimetic moieties containing peptide bonds, such as azole-derived mimetics, may be used as replacements for dipeptides. However, peptidomimetics and peptidomimetic backbones in which only amide bonds are replaced, as discussed above, are preferred.

[0078] Suitable peptidomimetics include reduced peptides in which the amide bonds have been reduced to methylene amines by treatment with a reducing agent, such as borane, or a hydride reagent, such as lithium aluminum hydride. Such reduction has the added benefit of increasing the cationicity of the overall molecule.

[0079] Other peptidomimetics include peptoids, formed, for example, by the stepwise synthesis of amide-functionalized polyglycines. Some peptidomimetic backbones are readily accessible from their peptide precursors, e.g., permethylated peptides; suitable methods are described in Ostresh, JM, et al., Proc. Natl. Acad. Sci. USA (1994) 91, 11138-11142. Strongly basic conditions favor N-methylation over O-methylation, resulting in methylation of some or all of the nitrogen atoms of the peptide bonds and the N-terminal nitrogen.

[0080] Preferred peptide bond substitutes include esters, polyamines and their derivatives, and substituted alkanes and alkenes, particularly aminomethyl and ketomethylene. Preferably, the N- and C-termini of the peptidomimetic may be modified as discussed herein.

[0081] In some embodiments, the compound comprising a disubstituted β-amino acid is a compound (ie, a peptide, peptidomimetic, or amino acid derivative) that includes a group of formula (VI):

[0082] [ka]

[0083] wherein X and N of formula (VI) have their normal valences and therefore become further substituted when they are attached to other parts of the compound, such as additional amino acids or N- or C-terminal capping groups; any two of R1, R2, R3, and R4 are hydrogen atoms and two are substituents, which may be the same or different, contain at least seven non-hydrogen atoms, are lipophilic, and contain a cyclic group that is not directly attached to either the α or β carbon atom but is optionally linked or fused to the cyclic group of the other substituent, wherein the combined total number of non-hydrogen atoms for the two substituents when the cyclic groups are fused is at least 12; and X represents O, C, N, or S.

[0084] When the cyclic groups of each moiety are fused, R 1~4 It will be appreciated that the minimum number of non-hydrogen atoms in the two groups of 12 for the combined total is arrived at by adding the minimum numbers for the unfused groups (7 + 7 = 14) and subtracting 2 therefrom since two of the non-hydrogen atoms effectively participate in ring formation in each group. Preferably, the cyclic groups of each moiety when fused together are 1~4 The combined total of non-hydrogen atoms in the two groups is 14. α or C β Complex fused and linked groups can be envisioned, such that the two groups attached to may contain more than one pair of fused cyclic groups, with or without an additional connecting bond between the substituents. Preferably, however, the two substituents are not fused or linked, as molecules in which these groups have the greatest movement flexibility are preferred.

[0085] The nitrogen atom in the group of formula (VI) may of course be C β or C α Preferably, the group R 1~4 Preferably, the main chain (NC β -C α The five atoms of -CX) are connected to each other only in a linear, not cyclic, fashion.

[0086] R 1~4 The substituents of R are generally lipophilic in nature, preferably uncharged, and preferably have no more than two, more preferably no more than one polar group. 1~4 Preferably, one or both of the substituents contains at least 8, more preferably at least 9 or 10 non-hydrogen atoms, e.g., 7 to 13, 7 to 12, 8 to 12, or 9 to 11 non-hydrogen atoms. These two substituents are preferably the same, simply for ease of synthesis. Preferably, the two substituents are R1 and R2 or R3 and R4, with R3 and R4 being most preferred.

[0087] As mentioned above, R 1~4 The cyclic group is spaced from the α or β carbon atom by a chain of 1 to 4, preferably 1 to 3 atoms, so that it is not directly attached to either of them. These linking atoms may include nitrogen and / or oxygen, but are typically carbon atoms, and preferably the linking atoms are unsubstituted. Preferred spacing moieties are shown in the examples and are R as defined herein. 1~4 It forms part of the substituent of

[0088] X may be substituted or unsubstituted, and is preferably an N atom, and is preferably substituted. When X is N, it may form part of an amide bond with a further amino acid. Alternatively, the N atom may be substituted with an aminoalkyl group, such as aminoethyl or aminopropyl or dimethylaminoethyl. In some embodiments, the N atom may form part of a cyclic group, such as piperazine, which itself may be substituted with an alkyl or aminoalkyl group.

[0089] Compounds including a group of formula (VI) will preferably have a modified C-terminus, which is preferably amidated.

[0090] The previous section defining preferred substituents for β-amino acids is 1~4The same also applies mutatis mutandis to the two substituents of formula VI. Because compounds incorporating the group of formula I are a preferred subset of the compounds described previously in this application, all of the preceding paragraphs defining compound characteristics, such as the length of the compounds and the other amino acids they contain, also apply to these compounds defined by the inclusion of the group of formula VI, and vice versa. Particularly preferred compounds are 1 to 7 or 8 (e.g., 1 to 5), more preferably 1, 2, 3, or 4 amino acids in length. Peptidomimetic molecules will contain the same number of subunits, but these subunits are typically linked by amide bond mimetics. Preferred linkages are discussed above and include esters and aminomethyl and ketomethylene.

[0091] In some embodiments, the compound comprising a disubstituted β-amino acid is a dipeptide (compounds 4a-4n) as shown in Figure 1 of WO 2011 / 051692 A1. In some other embodiments, the compound comprising a disubstituted β-amino acid is an amino acid derivative 1-14 and 23-38 shown in Table 2, Tables 4-7 of Example 2, and Figure 3 of WO 2011 / 051692 A1. In yet some other embodiments, the compound comprising a disubstituted β-amino acid is a heptapeptide 15-22 shown in Table 3 of Example 2 of WO 2011 / 051692 A1.

[0092] As discussed above, compounds containing disubstituted β-amino acids may be in the form of a salt, cyclic, or esterified, or may be amidated.

[0093] Compounds containing disubstituted β-amino acids may be synthesized in any convenient manner, which methods are known to those skilled in the art.

[0094] Compounds containing disubstituted β-amino acids and their synthesis are described in detail in WO 2011 / 051692 A1, the contents of which are incorporated herein by reference.

[0095] In a preferred embodiment, the compound comprising a disubstituted β-amino acid is LTX-401, i.e., a compound of formula (VII):

[0096] [ka]

[0097] It is preferably in the form of a pharmaceutically acceptable salt, more preferably in the form of its hydrochloride or acetate salt.

[0098] Chitosan and Chitosan Derivatives Chitosan is a linear polysaccharide composed of randomly distributed β-(1→4)-linked D-glucosamine (deacetylated units) and N-acetyl-D-glucosamine (acetylated units).

[0099] Chitosan is commercially produced by deacetylation of chitin, a structural component of the exoskeleton of crustaceans such as crabs and shrimp, but can also be produced from certain types of fungi and algae. Chitosan's key characteristics are its molecular weight and degree of acetylation (DA) or deacetylation (DDA), which correspond to the mole fraction of deacetylated and acetylated units. Most commercial chitosans have molecular weights ranging from <50 to 2000 kDa and average DDAs of 50 to 100%. Based on molecular weight, chitosans can be grouped into ultra-low molecular weight (typically <50 kDa), low molecular weight (typically 50 to <100 kDa), intermediate molecular weight (typically 100 to 1000 kDa), and high molecular weight (typically >1000 kDa). In some embodiments, the DDA of chitosan for use in the present disclosure is about 50-99%, e.g., 70-99%, e.g., 80%, preferably about 70% or more, e.g., 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0100] Chitosan for use in the present disclosure may be ultra-low molecular weight chitosan, low molecular weight chitosan, medium molecular weight chitosan, or high molecular weight chitosan, preferably ultra-low molecular weight chitosan, low molecular weight chitosan, or medium molecular weight chitosan.

[0101] Chitosan contains active functional groups that can undergo chemical reactions, and chitosan derivatives resulting from modifications of chitosan, such as acylation, carboxylation, alkylation, and quaternization, have properties that differ 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).

[0102] The chitosan derivative for use in the present disclosure may be a salt of the chitosan derivative, such as a pharmaceutically acceptable salt of chitosan, such as the hydrochloride salt. Hereinafter, the term "chitosan derivative" relates to the chitosan derivative and its pharmaceutically acceptable salts.

[0103] Chitosan derivatives for use in the present disclosure include O- and N-acylated chitosans, O- and N-alkylated chitosans, and chitosans 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.

[0104] In some embodiments, chitosan derivatives for use in the present disclosure are glycated chitosans 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, WO 2002 / 040055(A2), and WO 2013 / 109732(A2), the contents of which are incorporated herein by reference.

[0105] In some embodiments, glycated chitosans for use in the present disclosure have a percent glycation of about 0.1% to about 90% of their otherwise free amino groups, e.g., about 0.1% to about 30%, or e.g., about 2-15%, e.g., about 12.5%. In some embodiments, glycated chitosans for use in the present disclosure have a molecular weight of about 50 kD to about 2000 kD, e.g., about 50 kD to about 1500 kD, e.g., about 250 kD or 300 kD. In some embodiments, the DDA of glycated chitosans for use in the present disclosure is about 50-99%, e.g., 70-99%, e.g., 80%. In some embodiments, the DDA of the glycated chitosan for use in the present disclosure is about 50-99%, such as 70-99%, e.g., 80%, 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%, e.g., about 12.5%, and has a molecular weight of about 50 kD to about 2000 kD, e.g., about 50 kD to about 1500 kD, e.g., about 250 kD or about 300 kD.

[0106] Compositions containing oncolytic peptides The composition may comprise one oncolytic peptide or several different oncolytic peptides.

[0107] The composition comprising an oncolytic peptide is generally a composition comprising an oncolytic peptide and a physiologically acceptable carrier. In some embodiments, the composition is a pharmaceutical composition.

[0108] In some embodiments, the physiologically acceptable carrier is water, saline, or an aqueous buffer solution. In some embodiments, the physiologically acceptable carrier is water, e.g., water for injection (WFI). In some other embodiments, the physiologically acceptable carrier is water containing a salt, e.g., sodium chloride (saline). In yet some other embodiments, the physiologically acceptable carrier is an aqueous buffer solution or a mixture of several different aqueous buffer solutions. In some embodiments, the composition comprises a mixture of several different physiologically acceptable carriers.

[0109] A buffer may be used to maintain the pH of the composition within a desired range, preferably 4.0 to 7.4, more preferably 5.0 to 7.0, even more preferably 5.0 to 6.5, for example 6.0.

[0110] The composition comprising the oncolytic peptide is preferably a solution of the oncolytic peptide in a physiologically acceptable carrier, more preferably a solution of the oncolytic peptide in water, saline, or an aqueous buffer or a mixture of several different aqueous buffers.

[0111] In some embodiments, the composition contains the oncolytic peptide at a concentration of 0.1-30 mg / ml, e.g., 1-5 mg / ml or 1-30 mg / ml, preferably 4-25 mg / ml, or more preferably 6-20 mg / ml. The amounts of peptide referred to above refer to net peptide amounts, where applicable (i.e., when the oncolytic peptide is in fact a peptide), and refer to the free base as opposed to salt forms of the oncolytic peptide as defined herein.

[0112] Net peptide amount is the total amount of peptide obtained after peptide synthesis, corrected for its purity (analytical, e.g., determined by HPLC) and net peptide content (analytical, e.g., determined by elemental analysis). Net peptide content is the ratio of the desired peptide to counterions, residual water, and peptidic impurities. Net peptide (%) is calculated as follows: Net peptide content (%) x purity (%) 100%

[0113] The result gives the percentage of net peptide in the total amount of peptide.

[0114] In some embodiments, the composition comprises one or more additional pharmaceutically acceptable excipients, such as excipients well known in the art for use in pharmaceutical compositions / medical products comprising aqueous solutions of active ingredients.

[0115] In some embodiments, a composition comprising an oncolytic peptide may be a solution contained in a ready-for-use container, e.g., a sealed container. In other embodiments, the composition may be prepared just prior to its use / administration, e.g., by reconstituting, e.g., by dissolving, in a pharmaceutically acceptable carrier, an oncolytic peptide provided in solid form (e.g., lyophilized form).

[0116] In some embodiments, the composition comprises LTX-401 and a pharmaceutically acceptable carrier, hi some embodiments, the composition is a solution of LTX-401 in a pharmaceutically acceptable carrier contained in a ready-to-use sealed container.

[0117] Compositions containing 9-mer peptides For compositions comprising one or more 9-mer peptides, such compositions in some embodiments comprise one or more of the following aqueous buffers: acetate buffer, aspartate buffer, sodium benzoate buffer, benzoic acid buffer, carbonate buffer, citrate buffer, glucono-delta-lactone buffer, glycine buffer, glycine HCl buffer, histidine buffer, histidine HCl buffer, hydrobromide buffer, phosphate buffer (e.g., PBS), sodium succinate buffer, disodium succinate buffer, succinic acid buffer, sulfate buffer, tartrate buffer, and / or sodium tartrate buffer. Preferably, the aqueous buffer is acetate / citrate buffer, citrate buffer, and / or phosphate buffer.

[0118] For example, in some embodiments, a composition comprising a 9-mer peptide comprises one or more preservatives. The following table lists examples of preferred preservatives. Suitable concentration ranges for these agents when present in the composition are also listed.

[0119] [Table 2]

[0120] 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%.

[0121] In some embodiments, the composition comprising the 9-mer peptide comprises one or more chelating agents for chelating metal ions. The following table lists examples of preferred chelating agents. The preferred concentration ranges for these agents when present in the composition are also listed.

[0122] [Table 3]

[0123] EDTA = ethylenediaminetetraacetic acid, DTPA = diethylenetriamine pentaacetate

[0124] 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.

[0125] In some embodiments, the composition comprising the 9-mer peptide comprises one or more antioxidants. The following table lists examples of preferred antioxidants. The preferred concentration ranges of the antioxidants when present in the composition are also listed.

[0126] [Table 4]

[0127] A preferred antioxidant is thiourea.

[0128] In some embodiments, compositions comprising a 9-mer peptide include one or more tonicity modifiers, such as dextrose, glycerin, lactose, mannitol, potassium chloride, sodium chloride, and / or sorbitol, to achieve an isotonic composition that reduces pain experienced by a patient receiving the composition. Preferably, the tonicity modifier is sodium chloride.

[0129] In some embodiments, compositions containing 9-mer peptides contain one or more stabilizers that inhibit aggregation of the peptides. 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 in an amount of 1-8% w / w, preferably 2-7% w / w, e.g., 3-6% w / w.

[0130] In some embodiments, a composition comprising a 9-mer peptide comprises one or more viscosity enhancers, such as gum acacia, agar, cellulose derivatives such as methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose, calcium carboxymethylcellulose, sodium carboxymethylcellulose, gelatin, hyaluronic acid, maltodextrin, pectin, poloxamer 407, PEG, polyethylene oxide, povidone (PVP), starch such as corn starch and tapioca starch, tragacanth, and / or β-cyclodextrin.

[0131] By increasing the viscosity of the composition comprising the 9-mer peptide (compared to water), the dispersion of the peptide from the administration site can be effectively slowed down.When the composition is administered into a tumor, increasing the viscosity can extend the time that the composition remains in the tumor and does not leak out, slow down the dispersion of the peptide by blood, and increase the time that the peptide directly contacts tumor cells.In this way, the effectiveness of the peptide can be increased and / or toxicity can be reduced.

[0132] Thus, in some embodiments, a composition comprising a 9-mer peptide comprises one or more viscosity-enhancing agents to provide a viscosity suitable for injection, e.g., injection into a tumor. In some embodiments, the viscosity of the composition comprising an oncolytic peptide is from 1.5, 2, 2.5, or 3 cP to 300 cP, e.g., 2.5-250 cP, e.g., 5-100 cP or 10-200 cP and 20-240 cP, preferably 40-180 cP, e.g., 45-150 cP or 30-100 cP, more preferably 1.5-10 cP, e.g., 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.

[0133] The viscosity enhancing agent is preferably present at 0.4 to 1.2% w / w, for example 0.5 to 1% w / w.

[0134] Preferably, the one or more viscosity enhancing agents are selected from the group consisting of hydroxyethyl cellulose, hydroxypropyl cellulose, HPMC, poloxamer, PEG, and PVP.

[0135] When a composition comprising a 9-mer peptide is prepared prior to administration, excipients such as the preservatives, antioxidants, or stabilizers described above may not be necessary or, when present in the reconstituted composition, are preferably present in a pharmaceutically acceptable carrier. In some embodiments, when a composition is prepared prior to use / administration, any of the excipients described above are present in a pharmaceutically acceptable carrier.

[0136] In some embodiments, the composition comprises LTX-315 and a pharmaceutically acceptable carrier, preferably wherein the LTX-315 has the amino acid sequence of SEQ ID NO:23, or a pharmaceutically acceptable salt thereof, preferably the acetate salt thereof.

[0137] In some embodiments, such compositions are solutions of LTX-315 in a pharmaceutically acceptable carrier contained in a ready-to-use sealed container. In some other embodiments, such compositions are solutions of LTX-315 that are reconstituted prior to use / administration by dissolving such LTX-315 in solid form (e.g., as a lyophilized powder) in a pharmaceutically acceptable carrier.

[0138] Preparation of compositions containing oncolytic peptides In some embodiments, the composition comprising the oncolytic peptide is a pharmaceutical grade product. In some embodiments, the composition comprising the oncolytic peptide is a sterile composition. In some embodiments, the composition comprising the oncolytic peptide is a pharmaceutical composition.

[0139] Compositions containing oncolytic peptides may be prepared using any conventional method.

[0140] In some embodiments, the composition comprising the oncolytic peptide is a solution contained in a ready-to-use sealed container. To produce such a product, the oncolytic peptide in a solid form, for example, in the form of a (lyophilized) powder, may be dissolved in a pharmaceutically acceptable carrier under aseptic manufacturing conditions, and the resulting composition is preferably sterile filtered, then filled into a container and sealed. Alternatively, the oncolytic peptide may be dissolved in a pharmaceutically acceptable carrier, and the resulting composition is then filled into a container and sealed, and the container is then autoclaved to obtain a sterile composition. When excipients are present in the composition, depending on the nature and amount of the excipients, these excipients may be mixed or dissolved in the pharmaceutically acceptable carrier or in a solution containing the oncolytic peptide.

[0141] In some other embodiments, a composition comprising an oncolytic peptide is prepared immediately prior to use / administration, e.g., by dissolving, e.g., reconstituting, in a sterile pharmaceutically acceptable carrier, an oncolytic peptide provided in solid form (e.g., in the form of a lyophilized powder). To produce such a product, in some embodiments, the oncolytic peptide is dissolved, sterile filtered, and lyophilized. In some other embodiments, the oncolytic peptide is dissolved and lyophilized, and the lyophilized oncolytic peptide is sterilized by gamma irradiation.

[0142] Compositions containing chitosan and / or chitosan derivatives The composition may comprise one or more chitosans, for example several different chitosans, for example chitosans of different molecular weights and / or different DDAs, and / or may comprise one or more chitosan derivatives, for example several different chitosan derivatives, for example chitosan derivatives containing the same modifications but with different degrees of modification or containing different modifications.

[0143] The composition comprising chitosan and / or a chitosan derivative is generally a composition comprising chitosan and / or a chitosan derivative and a physiologically acceptable carrier. In some embodiments, the composition comprising chitosan and / or a chitosan derivative is a pharmaceutical composition.

[0144] In some embodiments, the composition comprising chitosan and / or chitosan derivatives is a dispersion, suspension, or solution of chitosan and / or chitosan derivatives in a physiologically acceptable carrier.

[0145] In some embodiments, the physiologically acceptable carrier is water, saline, or an aqueous buffer solution. In some embodiments, the physiologically acceptable carrier is water, e.g., water for injection (WFI). In some other embodiments, the physiologically acceptable carrier is water containing a salt, such as sodium chloride (saline) or other salts, e.g., potassium chloride, calcium chloride, or magnesium chloride, or the physiologically acceptable carrier is an aqueous buffer containing a salt, such as sodium chloride or other salts, e.g., potassium chloride, calcium chloride, or magnesium chloride. In some embodiments, the composition comprises a mixture of several different physiologically acceptable carriers. In yet some other embodiments, the physiologically acceptable carrier is an aqueous buffer solution or a mixture of several different aqueous buffer solutions.

[0146] A buffer may be used to maintain the pH of the composition within a desired range. Typically, the pH of the composition is 4.0 to 7.5. Preferably, the pH of the composition is 5.0 to 7.4, more preferably 5.0 to 7.0, e.g., 6.5 or 6.0.

[0147] In some embodiments, the composition comprising chitosan and / or a chitosan derivative has a viscosity suitable for injection, e.g., a viscosity suitable for injection into a subject's tumor. In some embodiments, the viscosity of the composition comprising chitosan and / or a chitosan derivative is 1.0, 1.5, 2, 2.5, or 3 cP to 300 cP, e.g., 2.5-250 cP, e.g., 5-100 cP or 10-200 cP and 20-240 cP, preferably 40-180 cP, e.g., 45-150 cP or 30-100 cP, more preferably 1.5-10 cP, e.g., 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. In some embodiments, the composition comprises glycated chitosan and has a viscosity of 1-100 cP.

[0148] In some embodiments, the composition comprises 0.1-5% w / w chitosan and / or chitosan derivatives, such as 0.1-2.5% w / w, for example 0.5%, 1%, 1.5%, or 2% w / w.

[0149] In some cases, the compositions include one or more additional pharmaceutically acceptable excipients, such as excipients well known in the art for use in medical products including aqueous dispersions, suspensions, or solutions of active ingredients.

[0150] In some embodiments, the composition comprises glycated chitosan, hi some other embodiments, the composition comprises galactocitosan.

[0151] Preparation of compositions containing chitosan and / or chitosan derivatives In some embodiments, the composition comprising chitosan and / or chitosan derivatives is a pharmaceutical grade product. In some embodiments, the composition comprising chitosan and / or chitosan derivatives is a sterile composition. In some embodiments, the composition comprising chitosan and / or chitosan derivatives is a pharmaceutical composition.

[0152] Compositions containing chitosan and / or chitosan derivatives may be manufactured using any conventional method. In some embodiments, chitosan and / or chitosan derivatives are dispersed, suspended, or dissolved in a physiologically acceptable carrier, the resulting composition is sterile filtered, and then filled into a container and sealed. In some embodiments, chitosan and / or chitosan derivatives are dispersed, suspended, or dissolved in a physiologically acceptable carrier, and the resulting composition is autoclaved. In some embodiments, compositions containing chitosan and / or chitosan derivatives are prepared immediately before use / administration, e.g., by dissolving, dispersing, or suspending chitosan and / or chitosan derivatives provided in solid form (e.g., lyophilized form) in a sterile, pharmaceutically acceptable carrier, e.g., by reconstitution. To manufacture such products, in some embodiments, chitosan and / or chitosan derivatives are dissolved, sterile filtered, and lyophilized. In some other embodiments, chitosan and / or chitosan derivatives are dissolved and lyophilized, and the lyophilized compound is sterilized by gamma irradiation.

[0153] Kit for compositions (a) and (b) Compositions (a) and (b) may be provided in the form of a kit that includes both such compositions or that includes parts / components for preparing compositions (a) and (b). In some embodiments, the kit includes compositions (a) and (b), each of which is in a (sealed) container.

[0154] The kit will typically include instructions for using compositions (a) and (b).

[0155] In some other embodiments, the kit comprises an oncolytic peptide and a physiologically acceptable carrier for preparing composition (a), and chitosan and / or a chitosan derivative and a physiologically acceptable carrier for preparing composition (b). In some embodiments, the physiologically acceptable carrier for preparing composition (a) is the same as the physiologically acceptable carrier for preparing composition (b). In such embodiments, the kit may comprise three containers or three compartments contained in one or two containers: one containing the oncolytic peptide, one containing chitosan and / or a chitosan derivative (e.g., both in solid form), and one containing a physiologically acceptable carrier for preparing compositions (a) and (b). In some other embodiments, the physiologically acceptable carriers for preparing compositions (a) and (b) are different. In such embodiments, the kit may include four containers or four compartments contained in one, two, or three containers, one containing an oncolytic peptide, one containing chitosan and / or a chitosan derivative (e.g., both in solid form), one containing a physiologically acceptable carrier for preparing composition (a), and one containing a physiologically acceptable carrier for preparing composition (b).

[0156] The kit will typically include instructions for preparing compositions (a) and (b), e.g., by reconstituting the oncolytic peptide and chitosan and / or chitosan derivative with a physiologically acceptable carrier prior to use, and instructions for using compositions (a) and (b).

[0157] In yet some other embodiments, the kit comprises composition (a), chitosan and / or chitosan derivatives (e.g., in solid form), and a physiologically acceptable carrier for preparing composition (b).

[0158] The kit will typically include instructions for preparing composition (b), e.g., by reconstituting chitosan and / or chitosan derivatives with a physiologically acceptable carrier prior to use, and instructions for using compositions (a) and (b).

[0159] In yet some other embodiments, the kit comprises composition (b), an oncolytic peptide (e.g., in solid form), and a physiologically acceptable carrier for preparing composition (a).

[0160] The kit will typically include instructions for preparing composition (a), e.g., by reconstituting the oncolytic peptide with a physiologically acceptable carrier prior to use, and instructions for using compositions (a) and (b).

[0161] A composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative A composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative may be referred to herein as a "combination composition."

[0162] The combination composition may comprise one oncolytic peptide or several different oncolytic peptides.Furthermore, the combination composition may comprise one or more chitosans, e.g., several different chitosans, e.g., chitosans of different molecular weights and / or different DDAs, and / or one or more chitosan derivatives, e.g., several different chitosan derivatives, e.g., chitosan derivatives containing the same modifications but with different degrees of modification or containing different modifications.

[0163] The combination composition is generally a composition comprising i) and ii) and a physiologically acceptable carrier. In some embodiments, the combination composition is a pharmaceutical composition.

[0164] In some embodiments, the combination composition is a dispersion, suspension, or solution, i.e., a dispersion, suspension, or solution of i) and ii) in a physiologically acceptable carrier. In some embodiments, the oncolytic peptide is dissolved in a physiologically acceptable carrier, while the chitosan and / or chitosan derivative is dispersed or suspended in the dissolved oncolytic peptide.

[0165] In some embodiments, the physiologically acceptable carrier is water, saline, or an aqueous buffer solution. In some embodiments, the physiologically acceptable carrier is water, e.g., water for injection (WFI). In some other embodiments, the physiologically acceptable carrier is water containing a salt, such as sodium chloride (saline) or other salts, e.g., potassium chloride, calcium chloride, or magnesium chloride, or the physiologically acceptable carrier is an aqueous buffer containing a salt, such as sodium chloride or other salts, e.g., potassium chloride, calcium chloride, or magnesium chloride. In yet some other embodiments, the physiologically acceptable carrier is an aqueous buffer solution or a mixture of several different aqueous buffer solutions.

[0166] A buffer may be used to maintain the pH of the combination composition within a desired range, preferably 4.0 to 7.4, more preferably 5.0 to 7.0, for example 6.0 or 6.5.

[0167] The combination composition contains the oncolytic peptide at a concentration of 0.1-30 mg / ml, e.g., 1-5 mg / ml or 1-30 mg / ml, preferably 4-25 mg / ml, or more preferably 6-20 mg / ml. The amounts of peptide mentioned above refer to the net peptide amount and the free base, where applicable (i.e., when the oncolytic peptide is in fact a peptide).

[0168] The combination composition comprises 0.1-5% w / w chitosan and / or chitosan derivatives, such as 0.1-2.5% w / w, such as 0.5% to 1.5%, such as 0.6% to 1% or 1% to 1.5% or 1.5% to 2% or 2% to 2.5% w / w.

[0169] In some embodiments, the combination composition has a viscosity suitable for injection, e.g., injection into a tumor in a subject. In some embodiments, the viscosity of the combination composition is from 1.0, 1.5, 2, 2.5, or 3 cP to 300 cP, e.g., 2.5-250 cP, e.g., 5-100 cP or 10-200 cP and 20-240 cP, preferably 40-180 cP, e.g., 45-150 cP or 30-100 cP, more preferably 1.5-10 cP, e.g., 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.

[0170] For combination compositions comprising a 9-mer peptide, such compositions may further comprise excipients as described in the respective sections herein, namely the "Compositions Comprising a 9-mer Peptide" section.

[0171] In some embodiments, the combination composition includes LTX-401. In some other embodiments, the combination composition includes LTX-315. In yet some other embodiments, the combination composition includes a saccharified chitosan, such as galactochitosan. In yet some other embodiments, the combination composition includes LTX-401 and a saccharified chitosan, such as galactochitosan, or includes LTX-315 and a saccharified chitosan, such as galactochitosan.

[0172] Preparation of the Combination Composition In some embodiments, the combination composition is a pharmaceutical grade product. In some embodiments, the combination composition is a sterile composition. In some embodiments, the combination composition is a pharmaceutical composition.

[0173] The combination composition may be prepared using any conventional method. In some embodiments, chitosan and / or chitosan derivatives are dispersed, suspended, or dissolved in a physiologically acceptable carrier, and the oncolytic peptide is dissolved in the dispersion, suspension, or solution. In some other embodiments, chitosan and / or chitosan derivatives are dispersed, suspended, or dissolved in a solution of the oncolytic peptide in a physiologically acceptable carrier. The solution of the oncolytic peptide may be obtained by dissolving the oncolytic peptide in a solid form, for example, in powder form, in a physiologically acceptable carrier. In some embodiments, the combination composition is sterile filtered, then filled into a container and sealed, or filled into a container and autoclaved.

[0174] Kits for Combination Compositions In the kit, parts / components for preparing the combination may be provided, ie the combination composition may be prepared prior to use / administration.

[0175] In some embodiments, the kit comprises compositions (a) and (b) described herein and instructions for preparing the combination composition. The kit will typically include instructions for using the combination composition.

[0176] In some other embodiments, the kit comprises an oncolytic peptide, chitosan and / or a chitosan derivative (e.g., both in solid form), and a physiologically acceptable carrier for preparing a combination composition. In such embodiments, the kit may comprise three containers or three compartments contained in one or two containers, one containing the oncolytic peptide, one containing chitosan and / or a chitosan derivative (e.g., both in solid form), and one containing a physiologically acceptable carrier for preparing a combination composition; or the kit may comprise two containers or two compartments contained in one container, one containing a mixture of the oncolytic peptide and chitosan and / or a chitosan derivative, and one containing a physiologically acceptable carrier for preparing a combination composition.

[0177] The kit will typically include instructions for preparing the combination composition, for example by reconstituting the oncolytic peptide and chitosan and / or chitosan derivative, or a mixture thereof, with a physiologically acceptable carrier before use, and instructions for using the combination composition.

[0178] In yet some other embodiments, the kit comprises composition (a) and chitosan and / or a chitosan derivative (eg, in solid form).

[0179] The kit will typically include instructions for preparing the combination composition, for example by reconstituting chitosan and / or chitosan derivatives with composition (a) before use, and instructions for using the combination composition.

[0180] In yet some other embodiments, the kit comprises composition (b) and an oncolytic peptide (eg, in solid form).

[0181] The kit will typically include instructions for preparing the combination composition, for example by reconstituting the oncolytic peptide with composition (b) prior to use, and instructions for using the combination composition.

[0182] Methods of treating a subject with a tumor / Use of the compositions disclosed herein in methods of treating a subject with a tumor Compositions (a) and (b) disclosed herein are used simultaneously, sequentially, or as a combined composition (i.e., a combined composition disclosed herein) in a method of treating a subject having a tumor, e.g., are administered to the subject simultaneously, sequentially, or as a combined composition.

[0183] In some embodiments, compositions (a) and (b) are administered simultaneously, i.e., at the same time. In this context, concurrently means simultaneously (e.g., by the same or different administration methods) or within a short period of time, e.g., minutes or hours or more, but on the same day. In some embodiments, compositions (a) and (b) are administered simultaneously, wherein composition (b) comprising chitosan and / or a chitosan derivative is administered first, followed by composition (a) comprising an oncolytic peptide. In some other embodiments, compositions (a) and (b) are administered simultaneously, wherein composition (a) comprising an oncolytic peptide is administered first, followed by composition (b) comprising chitosan and / or a chitosan derivative.

[0184] In some other embodiments, compositions (a) and (b) are administered sequentially. In this context, sequential means, for example, on different days, one day / days or one week / weeks apart, using the same or different administration methods. In some embodiments, compositions (a) and (b) are administered sequentially, where composition (b) comprising chitosan and / or a chitosan derivative is administered first, followed by composition (a) comprising an oncolytic peptide. In some other embodiments, compositions (a) and (b) are administered sequentially, where composition (a) comprising an oncolytic peptide is administered first, followed by composition (b) comprising chitosan and / or a chitosan derivative.

[0185] In some embodiments, compositions (a) and (b) are each administered only once, i.e., one dose is administered at one time point during the course of treatment. In some other embodiments, compositions (a) and (b) are each administered repeatedly, i.e., several doses are administered at several time points during the course of treatment. In some other embodiments, one of compositions (a) and (b) is administered repeatedly, while the other is administered only once.

[0186] In some embodiments, the composition is a combination composition. In some embodiments, such a combination composition is administered only once. In some other embodiments, such a combination composition is administered repeatedly over the course of treatment.

[0187] The combined composition and composition (a) are preferably administered intratumorally. In some embodiments, the composition is injected into the tumor. In some other embodiments, the composition is administered intratumorally by perfusion / infusion of a preferably isolated (including partially isolated) limb, body region, or organ containing the tumor. Any one or a combination of both modes of administration (injection and perfusion / infusion) to localize the composition to the tumor site is acceptable, as long as the delivery mechanism ensures sufficient concentration of the components of the composition in the tumor.

[0188] Composition (b) is preferably administered intratumorally and / or into the tissue immediately surrounding the tumor. As described above for composition (a) and the combination composition, composition (b) may be administered by injection, perfusion / infusion, or a combination of both methods.

[0189] In the methods of treatment / use of the compositions disclosed herein for treatment, the compositions are preferably administered in a therapeutically effective amount, which 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, for example, several days, weeks, or months.

[0190] The actual dose administered may vary and will depend on the age, weight, sex, medical history, past medical history, and general condition of the subject, the severity of the condition being treated, and the judgment of a health care professional.

[0191] The method of treatment / use of the compositions disclosed herein for treatment may continue as long as the clinician overseeing the patient's care deems the method effective and the treatment necessary.

[0192] In some embodiments, the methods of treatment / use of the compositions disclosed herein for treatment are for treating tumors selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, bone cancer, colorectal cancer, stomach cancer, skin cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, thyroid cancer, kidney cancer, liver cancer, bile duct cancer, brain cancer, cervical cancer, bladder cancer, esophageal cancer, Hodgkin's disease, and adrenocortical carcinoma tumors. In some embodiments, the tumor is a carcinoma, adenocarcinoma, lymphoma, melanoma, blastoma, leukemia, sarcoma, and germ cell tumor. [Example]

[0193] The description set forth above is believed to be sufficient to enable one skilled in the art to practice the present invention. The following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way. Indeed, various modifications of the present invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and will fall within the scope of the appended claims. [Example]

[0194] Preparation of the compositions disclosed herein The following compositions were prepared as follows:

[0195] Approximately half of the aqueous acetic acid / acetate buffer or saline solution was added to a reaction vessel equipped with a magnetic stirring means and stirred. LTX-401 in the acetate salt form (prepared as described in WO 2011 / 051692(A1)) as a dry powder was added and stirred until dissolved. Chitosan or HPMC, if present in the composition, was added with stirring. The remaining buffer or saline solution was then added, and the mixture was vigorously stirred until the polymer was completely dissolved. All compositions were autoclaved at 121°C for 15 minutes.

[0196] [Table 5]

[0197] * 24 mg / ml corresponds to 20 mg / ml of the free base, and 12 mg / ml corresponds to 10 mg / ml of the free base. ntd: not determined CS LMW = chitosan low molecular weight, 340 kDa, DAA 85.4% (manufacturer: Chitinor, Norway) CS MMW = chitosan medium molecular weight, 350-600 kDa, DAA 88% (manufacturer: Chitinol, Norway) HPMC LMW = Hydroxypropyl methylcellulose low molecular weight, pharmaceutical grade HPMC metolose 60SH (LMW) (Safic Alcan, Paris La Defense Cedex, France) HPMC HMW = Hydroxypropyl methylcellulose high molecular weight, pharmaceutical grade HPMC Metrose 60SH (HMW) (Sufiq Alcan, Paris-La Défense-Cedex, France)

[0198] Viscosity was determined with a Fungilab / Evo expert viscometer with an LCP spindle. Measurements were performed at 25 °C, a speed of 10 rpm and 12.2 s for a sample volume of 20 mL. -1 and the pH was determined according to Ph.Eur. [Example]

[0199] In vitro cytotoxic activity The in vitro cytotoxic (oncolytic) activity of each of Compositions 1 to 15 in the table above was examined in three cell lines by determining their IC50 (half maximal inhibitory concentration), i.e., the concentration required to kill 50% of cells. Cell viability was determined by the WST-1 assay (Roche). RPMI 1640 medium without FCS (assay medium) and assay medium containing 1% v / v Triton X-100 were used as negative and positive controls, respectively. Three independent experiments per cell line were performed. Stock solutions of Compositions 1 to 15 containing 0.8 mg / ml LTX-401 were prepared and diluted to 2 to 200 μg / ml in assay medium.

[0200] The three cell lines used in this example were (all obtained from ATCC, USA): 143B, a human osteosarcoma cell line K7M2, a mouse osteosarcoma cell line B16F10, a mouse melanoma cell line

[0201] Cells were plated in 100 μl aliquots per well into 96-well flat-bottom microtitration plates and incubated in RPMI plus FCS at 37°C for 16 hours. Cells were washed once with RPMI without FCS, and 100 μl / well of diluted Compositions 1–15, negative control, or positive control was added. The plates were incubated at 37°C for 4 hours, after which time 10 μl of WST-1 solution was added to each well. The plates were incubated for an additional 2 hours. 70 μl of the solution from each well was removed, and 100 μl of acidified isopropanol was added. The optical density (OD) of each well was measured at 590 nm using a Varioskan™ LUX multimode microplate reader (Thermo Scientific). To assess the effect of HPMC or chitosan alone on cell viability, the cell viability of Compositions 1–8 and 10–14 without LTX-401 was determined by the WST-1 assay. None of these compositions affected cell viability.

[0202] Compositions 10, 12, 13, and 15 were selected for use in Example 3 based on their IC50 in three different cell lines, their LTX-401 concentration, and their viscosity. [Example]

[0203] In vivo cytotoxic activity against osteosarcoma The following compositions 10, 12, 13, and 15 were selected for in vivo testing in a syngeneic osteosarcoma mouse model by examining the effect of administering the compositions intratumorally. 10:LTX-401, 10mg / ml (free base), 0.5%w / w HPMC HMW 12: LTX-401, 10 mg / ml (free base), 1% w / w chitosan MMW 13: LTX-401, 10 mg / ml (free base), 1.2% w / w chitosan MMW 15: LTX-401, 10 mg / ml (free base) Control: Saline solution

[0204] Balb / c mice (Taconic Biosciences) were inoculated with 8 × 10 6 K7M2 tumor cells were inoculated on day 0 by subcutaneous injection of 100 μl of tumor cells. Mice were divided into five groups of seven mice each, and treatment began when tumors reached the desired size of approximately 5 mm in diameter. Each composition was administered by injection into the tumor (50 μl) for three consecutive days. Mice were monitored throughout the study by measuring tumors and weighing the animals periodically. Mice were followed until the maximum tumor volume reached a volume of 1.5 ml or a length of 20 mm, or a severe adverse event (i.e., wound formation or ulceration at the tumor / injection site) occurred, at which point they were sacrificed. Tumor size was measured using a caliper, and weighing and physical examination served as health controls.

[0205] The results are shown in Figure 1. Tumor growth in mice with the LTX-401 composition was slowed compared to tumor growth in mice receiving saline alone. Essentially no difference in tumor growth was observed between the group of mice treated with LTX-401 alone and the group of mice treated with LTX-401 / 0.5% HPMC. Mice treated with a composition containing LTX-401 and chitosan showed significantly slower tumor growth; after 150 days, one mouse in the group receiving LTX-401 / 1% chitosan and two mice in the group receiving LTX-401 / 1.2% chitosan were still alive and in remission (no visible or palpable tumors). [Example]

[0206] In vivo cytotoxic activity against melanoma The following compositions were prepared in aqueous acetic acid / acetate buffer, pH 5.2, as described in Example 1. The following chitosans were used in this study: Chitosan medium molecular weight, 330 kDa, DAA 86% (manufacturer Chitinol, Norway). 1:LTX-401, 5mg / ml (free base) 2: Chitosan, 1.2% (w / w) 3: LTX-401, 5 mg / ml (free base) and chitosan, 1.2% (w / w) 4: Control: Aqueous acetic acid / acetate buffer

[0207] Immunocompetent C57BL / 6NRj mice (JanVier Labs, France) were inoculated with 5 × 10 IgG antibodies per mouse onto the abdominal skin. 4 Animals were inoculated with B16F1 melanoma cells on day 0 by subcutaneous injection of 10 tumor cells / 50 μl RPMI-1640. Mice were divided into four groups of 10 mice each, and tumors were inoculated until they were palpable and approximately 5 mm in diameter (40–80 mm). 3 Treatment began when tumors reached the desired size (860-1100 mm). Each of Compositions 1-4 was administered by injection (50 μl) into the tumor for two consecutive days (days 11 and 12, or days 12 and 13, or days 13 and 14, depending on tumor growth). Mice were monitored throughout the study by measuring tumors and weighing the animals periodically. Tumor volume was measured using electronic calipers and expressed as volume V = (L × W × W) / 2, where W is the tumor width and L is the tumor length. Maximum tumor volume was approximately 12-113 mm in diameter (860-1100 mm). 3 Mice were followed until they reached a tumor size of 1000 mg / kg, metastasis occurred, or a severe adverse event (i.e., wound formation or ulceration at the tumor / injection site) occurred. Mice were sacrificed upon reaching any of these endpoints. A score sheet was used to assess the overall health of the animals based on parameters such as weight and physical examination.

[0208] The results are shown in Figure 2. The survival plot shows that Composition 3 (the combination composition of LTX-401 and chitosan) induced complete tumor regression in all treated animals, with a survival rate of 100%, compared to 60% for animals treated with LTX-401 alone and 10% for animals treated with chitosan alone. No animals in the control group survived.

[0209] Embodiment 1. A combination of a composition (a) comprising an oncolytic peptide and a composition (b) comprising chitosan and / or a chitosan derivative for use in treating a subject with a tumor.

[0210] 2. A composition (a) comprising an oncolytic peptide and a composition (b) comprising chitosan and / or a chitosan derivative for use in treating a subject having a tumor.

[0211] 3. A composition (a) comprising an oncolytic peptide for use in treating a subject having a tumor by simultaneous, sequential or combined administration with a composition (b) comprising chitosan and / or a chitosan derivative.

[0212] 4. A method for treating a subject having a tumor, comprising administering to the subject a composition (a) comprising an oncolytic peptide and a composition (b) comprising chitosan and / or a chitosan derivative.

[0213] 5. Use of a composition (a) comprising an oncolytic peptide and a composition (b) comprising chitosan and / or a chitosan derivative for treating a subject having a tumor.

[0214] 6. The combination, composition for use, method, or use according to any of embodiments 1 to 5, wherein said compositions (a) and (b) are administered to said subject, said administration being simultaneous, sequential, or combined administration.

[0215] 7. The combination, composition for use, method, or use according to any of embodiments 1 to 6, wherein said composition (a) comprises one or more oncolytic peptides and / or said composition (b) comprises one or more different chitosans and / or chitosan derivatives.

[0216] 8. The combination, composition for use, method, or use according to any of embodiments 1 to 7, wherein said composition (a) and / or said composition (b) comprises a physiologically acceptable carrier.

[0217] 9. The combination, composition for use, method, or use according to embodiment 8, wherein the physiologically acceptable carrier is selected from the group consisting of water, e.g., water for injection (WFI), saline, and aqueous buffer solutions.

[0218] 10. The combination, composition for use, method or use according to any of embodiments 8-9, wherein said composition (a) and / or said composition (b) comprise a mixture of different physiologically acceptable carriers.

[0219] 11. The combination, composition for use, method, or use according to any of embodiments 8-10, wherein said composition (a) and / or said composition (b) comprise water, e.g., water for injection, which water comprises a salt selected from the group consisting of sodium chloride, potassium chloride, calcium chloride, and magnesium chloride.

[0220] 12. The combination, composition for use, method, or use according to any of embodiments 1 to 11, wherein the oncolytic peptide is a 9-mer peptide consisting of 9 amino acids in a linear configuration, or a pharmaceutically acceptable salt thereof, wherein 5 of the 9 amino acids are cationic and 4 are lipophilic, and 3 of the 4 lipophilic amino acids are tryptophan and 1 is a non-genetically encoded amino acid.

[0221] 13. The combination, composition for use, method, or use according to embodiment 12, wherein the lipophilic and cationic amino acids are arranged such that no more than two of either type of amino acid are adjacent to one another and / or such that the peptide comprises two pairs of adjacent cationic amino acids and one or two pairs of adjacent lipophilic amino acids.

[0222] 14. The combination, composition for use, method, or use according to any of embodiments 12-13, wherein the cationic amino acids are the same or different and are selected from the group consisting of lysine, arginine, histidine, and non-genetically encoded amino acids that have a positive charge at pH 7.0, preferably selected from the group consisting of lysine and arginine.

[0223] 15. The combination, composition for use, method, or use according to any of embodiments 12 to 14, wherein the non-genetically encoded cationic amino acid is selected from the group consisting of derivatives of lysine, arginine, and histidine.

[0224] 16. The combination, composition for use, method, or use according to any of embodiments 12 to 15, wherein the non-genetically encoded cationic amino acid is selected from the group consisting of homolysine, ornithine, diaminobutyric acid, diaminopimelic acid, diaminopropionic acid, homoarginine, trimethylysine, trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-carbamimidoylpiperidine-4-carboxylic acid, and 4-guanidinophenylalanine.

[0225] 17. The combination, composition for use, method, or use according to any of embodiments 12-16, wherein the non-genetically encoded lipophilic amino acid has a lipophilic side chain having at least 7, at least 8, at least 9, or at least 10 non-hydrogen atoms, preferably said non-hydrogen atoms being carbon atoms.

[0226] 18. The combination, composition for use, method or use according to any of embodiments 12-17, wherein the non-genetically encoded lipophilic amino acid has a lipophilic side chain containing not more than 30 or not more than 25 non-hydrogen atoms, and / or such lipophilic side chain comprises at least one, preferably two, optionally fused or connected cyclic groups, and / or said lipophilic side chain contains a heteroatom, such as O, N or S, preferably only one heteroatom, more preferably nitrogen, and / or said lipophilic side chain comprises two or less than two polar groups, preferably one, more preferably no polar groups.

[0227] 19. Non-genetically encoded lipophilic amino acids include 2-amino-3-(biphenyl-4-yl)propanoic acid (biphenylalanine), 2-amino-3,3-diphenyl-propanoic 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-indol-3-yl)propanoic acid, 2-amino-3-[1,1':3',1"-terphenyl-4-yl]- 19. The combination, composition for use, method, or use according to any of embodiments 12 to 18, wherein the amino acid is selected from the group consisting of 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, more preferably selected from the group consisting of diphenylalanine and biphenylalanine.

[0228] 20. The combination, composition for use, method, or use according to any of embodiments 12-19, wherein the 9-mer peptide is a compound of formula (I) to (V), wherein C represents a cationic amino acid, L represents a lipophilic amino acid, and the amino acids are covalently linked by peptide bonds, preferably amidated at the carboxy terminus. CCLLCCLLC(I), LCCLLCCLC(II), CLLCCLLCC(III), CCLLCLLCC(IV), CLCCLLCCL(V).

[0229] 21. The combination, composition for use, method, or use according to any of embodiments 12-20, wherein the 9-mer peptide is a compound of formula (I'), (I''), (I''') or (II'), wherein C represents a cationic amino acid and L' represents a non-genetically encoded lipophilic amino acid, the amino acids are covalently linked by peptide bonds, and preferably are amidated at the carboxy terminus. CCL'LCCLLC(I'), CCLLCCLL'C(I''), CCLL'CCLLC(I'''), LCCLL'CCLC(II').

[0230] 22. Oncolytic peptides include LTX-301, LTX-302, LTX-303, LTX-304, LTX-305, LTX-306, LTX-307, LTX-308, LTX-309, LTX-310, L TX-310, LTX-311, LTX-312, LTX-313, LTX-314, LTX-315, LTX-316, LTX-317, LTX-318, LTX-319, LTX-320, LTX-321, L 22. The combination, composition for use, method, or use according to any of embodiments 1 to 21, wherein the compound is selected from the group consisting of LTX-322, LTX-323, LTX-324, LTX-325, LTX-326, LTX-327, LTX-328, LTX-329, LTX-330, LTX-331, LTX-332, LTX-333, LTX-334, LTX-335, LTX-336, LTX-337, and LTX-338.

[0231] 23. The combination, composition for use, method, or use according to any of embodiments 1 to 22, wherein the oncolytic peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329, preferably the oncolytic peptide is LTX-315.

[0232] 24. The combination, composition for use, method, or use according to any of embodiments 1 to 23, wherein the oncolytic peptide is LTX-315 in the form of a pharmaceutically acceptable salt, preferably the acetate salt.

[0233] 25. The combination, composition for use, method, or use according to any of embodiments 1-24, wherein composition (a) further comprises one or more preservatives and / or one or more chelating agents and / or one or more antioxidants and / or one or more tonicity adjusting agents and / or one or more stabilizers and / or one or more viscosity enhancing agents.

[0234] 26. The combination, composition for use, method, or use according to any of embodiments 1-11, wherein the oncolytic peptide is a peptide, peptidomimetic, or amino acid derivative having a net positive charge of at least +2 and comprising disubstituted β-amino acids, each substituent in the β-amino acids containing at least 7 non-hydrogen atoms, being lipophilic, and having at least one cyclic group, wherein one or more cyclic groups in a substituent are optionally fused with one or more cyclic groups in another substituent.

[0235] 27. The combination, composition for use, method, or use according to embodiment 26, wherein one or more cyclic groups in the substituents are fused with one or more cyclic groups in the other substituents, and the combined total number of non-hydrogen atoms for the two substituents is at least 12.

[0236] 28. The oncolytic peptide is a compound comprising a group of formula (VI):

[0237] [ka]

[0238] 28. The combination, composition for use, method, or use according to any of embodiments 26-27, wherein X and N have their normal valences and are bonded to the rest of the compound; any two of R, R, R, and R are hydrogen atoms and two are substituents, which substituents contain at least 7 non-hydrogen atoms, are lipophilic, and comprise a cyclic group, which cyclic group is not directly attached to either the α or β carbon atom and is optionally linked or fused to the cyclic group of the other substituent; and X represents O, C, N, or S.

[0239] 29. The combination, composition for use, method, or use according to embodiment 28, wherein said substituents are the same or different.

[0240] 30. The combination, composition for use, method, or use according to either embodiment 28 or 29, wherein said cyclic group is linked or fused to a cyclic group of the other substituent, and the combined total number of non-hydrogen atoms for the two substituents is at least 12.

[0241] 31. The nitrogen atom of the group of formula (VI) is a group R 1~4 and / or the main chain (NC β -C α -CX) five atoms are connected to each other in a linear fashion, and / or R 1~4 is lipophilic in nature, preferably uncharged, preferably has not more than two, more preferably not more than one polar group, and / or X is a substituted N atom.

[0242] 32. The combination, composition for use, method, or use according to any of embodiments 28 or 31, wherein the compound including a group of formula VI has an amidated C-terminus.

[0243] 33. The combination, composition for use, method, or use according to any of embodiments 1-11 and 26-32, wherein the oncolytic peptide is LTX-401, i.e., a compound of formula (VII).

[0244] [ka]

[0245] 34. The combination, composition for use, method or use according to embodiment 33, wherein LTX-401 is in the form of a pharmaceutically acceptable salt, preferably in the form of a hydrochloride or acetate salt.

[0246] 35. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (a) comprises the oncolytic peptide at a concentration of 0.1 to 30 mg / ml, such as 1 to 5 mg / ml or 1 to 30 mg / ml, preferably 4 to 25 mg / ml, or more preferably 6 to 20 mg / ml.

[0247] 36. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein the pH of composition (a) is 4.0 to 7.4, more preferably 5.0 to 7.0, even more preferably 5.0 to 6.5, for example 6.0.

[0248] 37. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein the oncolytic peptide is in the form of a pharmaceutically acceptable salt, more preferably in the form of a hydrochloride or acetate salt.

[0249] 38. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) comprises chitosan.

[0250] 39. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) comprises several different chitosans.

[0251] 40. The combination, composition for use, method, or use according to any of embodiments 38-39, wherein the chitosan is an ultra-low molecular weight chitosan, a low molecular weight chitosan, and / or an intermediate molecular weight chitosan.

[0252] 41. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) comprises a chitosan derivative.

[0253] 42. The combination, composition for use, method, or use according to embodiment 41, wherein composition (b) comprises several different chitosan derivatives.

[0254] 43. The combination, composition for use, method, or use according to any of embodiments 41-42, wherein the chitosan derivative is a glycated chitosan, preferably having a glycation rate of from about 0.1% to about 90% of its otherwise free amino groups and / or a molecular weight of from about 50 kD to about 2000 kD.

[0255] 44. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein the degree of deacetylation (DDA) of the chitosan and / or chitosan derivative is about 50-99%, preferably about 70% to about 99%.

[0256] 45. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) comprises 0.1-5% w / w of chitosan and / or chitosan derivatives, e.g., 0.1-2.5% w / w, e.g., 0.5%, 1%, 1.5%, or 2% w / w.

[0257] 46. ​​The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) is a dispersion, suspension, or solution of chitosan and / or chitosan derivatives in a physiologically acceptable carrier.

[0258] 47. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein the pH of composition (b) is 4.0 to 7.5, preferably 5.0 to 7.4, more preferably 5.0 to 7.0, such as 6.5 or 6.0.

[0259] 48. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein the viscosity of composition (a) and / or (b) is from 1.0, 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.

[0260] 49. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (a) is administered intratumorally, e.g., by injection and / or perfusion / infusion.

[0261] 50. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (b) is administered intratumorally, e.g., by injection and / or perfusion / infusion, and / or into the tissue immediately surrounding the tumor.

[0262] 51. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (a) and composition (b) are administered simultaneously, e.g., simultaneously (e.g., by the same or different dosing regimen), or within a short period of time, e.g., minutes or hours or more, but on the same day.

[0263] 52. The combination, composition for use, method, or use according to embodiment 51, wherein composition (a) is administered first, followed by composition (b).

[0264] 53. The combination, composition for use, method, or use according to embodiment 51, wherein composition (b) is administered first, followed by composition (a).

[0265] 54. A combination, composition for use, method, or use according to any of embodiments 1-50, wherein composition (a) and composition (b) are administered sequentially, e.g., one or several days apart, or on different days, e.g., one or several weeks apart (e.g., with the same or different administration regimes).

[0266] 55. The combination, composition for use, method, or use according to embodiment 54, wherein composition (a) is administered first, followed by composition (b).

[0267] 56. The combination, composition for use, method, or use according to embodiment 54, wherein composition (b) is administered first, followed by composition (a).

[0268] 57. A combination, composition for use, method, or use according to any of the preceding embodiments, wherein each of compositions (a) and b) is administered only once, e.g., administered intratumorally.

[0269] 58. The combination, composition for use, method, or use according to any of embodiments 1-56, wherein each of compositions (a) and (b) is administered repeatedly, e.g., intratumorally.

[0270] 59. The combination, composition for use, method, or use according to any of embodiments 1-56, wherein one of compositions (a) and (b) is administered only once, e.g., intratumorally, and the other composition is administered repeatedly, e.g., intratumorally.

[0271] 60. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (a) and composition (b) are administered, e.g., administered intratumorally, in therapeutically effective amounts, and such amounts are administered in one administration or several administrations.

[0272] 61. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein said tumor is a tumor selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, bone cancer, colorectal cancer, gastric cancer, skin cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, thyroid cancer, kidney cancer, liver cancer, cancer of the bile duct, brain cancer, cervical cancer, bladder cancer, esophageal cancer, Hodgkin's disease, and adrenocortical carcinoma tumors.

[0273] 62. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein said tumor is a tumor selected from the group consisting of carcinoma, adenocarcinoma, lymphoma, melanoma, blastoma, leukemia, sarcoma, and germ cell tumor.

[0274] 63. The combination, composition for use, method, or use according to any of the preceding embodiments, wherein composition (a) and composition (b) are pharmaceutical compositions.

[0275] 64. A composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative.

[0276] 65. The composition according to embodiment 64, comprising one or more oncolytic peptides and / or one or more different chitosans and / or chitosan derivatives.

[0277] 66. The composition according to any one of embodiments 64-65, wherein the composition comprises a physiologically acceptable carrier.

[0278] 67. The composition according to embodiment 66, wherein the physiologically acceptable carrier is selected from the group consisting of water, e.g., water for injection (WFI), saline, and aqueous buffer solutions.

[0279] 68. The composition according to any of embodiments 66-67, wherein the composition comprises a mixture of different physiologically acceptable carriers.

[0280] 69. The composition according to any of embodiments 66-68, wherein the composition comprises water, e.g., water for injection, and contains a salt selected from the group consisting of sodium chloride, potassium chloride, calcium chloride, and magnesium chloride.

[0281] 70. The composition according to any of embodiments 64-69, wherein the oncolytic peptide is a 9-mer peptide consisting of 9 amino acids in a linear configuration, or a pharmaceutically acceptable salt thereof, wherein 5 of the 9 amino acids are cationic and 4 are lipophilic, and 3 of the 4 lipophilic amino acids are tryptophan and 1 is a non-genetically encoded amino acid.

[0282] 71. The composition according to embodiment 70, wherein the lipophilic and cationic amino acids are arranged so that no more than two of either type of amino acid are adjacent to each other, and / or such that the peptide contains two pairs of adjacent cationic amino acids and one or two pairs of adjacent lipophilic amino acids.

[0283] 72. The composition according to any of embodiments 70-71, wherein the cationic amino acids are the same or different and are selected from the group consisting of lysine, arginine, histidine, and non-genetically encoded amino acids that have a positive charge at pH 7.0, preferably selected from the group consisting of lysine and arginine.

[0284] 73. The composition according to any of embodiments 70-72, wherein the non-genetically encoded cationic amino acid is selected from the group consisting of derivatives of lysine, arginine, and histidine.

[0285] 74. The composition according to any of embodiments 70-73, wherein the non-genetically encoded cationic amino acid is selected from the group consisting of homolysine, ornithine, diaminobutyric acid, diaminopimelic acid, diaminopropionic acid, homoarginine, trimethylysine, trimethylornithine, 4-aminopiperidine-4-carboxylic acid, 4-amino-1-carbamimidoylpiperidine-4-carboxylic acid, and 4-guanidinophenylalanine.

[0286] 75. The composition according to any of embodiments 70-74, wherein the non-genetically encoded lipophilic amino acid has a lipophilic side chain having at least 7, at least 8, at least 9, or at least 10 non-hydrogen atoms, preferably said non-hydrogen atoms are carbon atoms.

[0287] 76. The composition according to any of embodiments 70-75, wherein the non-genetically encoded lipophilic amino acid has a lipophilic side chain containing not more than 30 or not more than 25 non-hydrogen atoms, and / or such lipophilic side chain comprises at least one, preferably two, optionally fused or connected cyclic groups, and / or said lipophilic side chain contains a heteroatom, such as O, N, or S, preferably only one heteroatom, more preferably nitrogen, and / or said lipophilic side chain comprises two or less than two polar groups, preferably one, more preferably no polar groups.

[0288] 77. Non-genetically encoded lipophilic amino acids include 2-amino-3-(biphenyl-4-yl)propanoic acid (biphenylalanine), 2-amino-3,3-diphenyl-propanoic 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-indol-3-yl)propanoic acid, 2-amino-3-[1,1':3',1"-terfen-4-yl]-propionic acid, 2-amino-3-(2,5,7-tri-tert-butyl-1H-indol-3-yl)propanoic acid, and 2-amino-3-[1,1':3',1"-terfen-4-yl]-propionic acid. 77. The composition according to any of embodiments 70-76, wherein the amino acid residue is selected from the group consisting of 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, more preferably selected from the group consisting of diphenylalanine and biphenylalanine.

[0289] 78. The composition according to any of embodiments 70-77, wherein the 9-mer peptide is a compound of formula (I) to (V), wherein C represents a cationic amino acid, L represents a lipophilic amino acid, and the amino acids are covalently linked by peptide bonds, preferably amidated at the carboxy terminus. CCLLCCLLC(I), LCCLLCCLC(II), CLLCCLLCC(III), CCLLCLLCC(IV), CLCCLLCCL(V).

[0290] 79. The composition according to any of embodiments 70-78, wherein the 9-mer peptide is a compound of formula (I'), (I''), (I''') or (II'), wherein C represents a cationic amino acid and L' represents a non-genetically encoded lipophilic amino acid, the amino acids are covalently linked by peptide bonds, and preferably are amidated at the carboxy terminus. CCL'LCCLLC(I'), CCLLCCLL'C(I''), CCLL'CCLLC(I'''), LCCLL'CCLC(II').

[0291] 80. Oncolytic peptides include LTX-301, LTX-302, LTX-303, LTX-304, LTX-305, LTX-306, LTX-307, LTX-308, LTX-309, LTX- 310, LTX-310, LTX-311, LTX-312, LTX-313, LTX-314, LTX-315, LTX-316, LTX-317, LTX-318, LTX-319, LTX-320 80. The composition according to any of embodiments 70-79, wherein the compound is selected from the group consisting of LTX-321, LTX-322, LTX-323, LTX-324, LTX-325, LTX-326, LTX-327, LTX-328, LTX-329, LTX-330, LTX-331, LTX-332, LTX-333, LTX-334, LTX-335, LTX-336, LTX-337, and LTX-338.

[0292] 81. The composition according to any of embodiments 70-80, wherein the oncolytic peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329, preferably the oncolytic peptide is LTX-315.

[0293] 82. The composition according to any of embodiments 70-81, wherein the oncolytic peptide is LTX-315 in the form of a pharmaceutically acceptable salt, preferably in the form of the acetate salt.

[0294] 83. The composition according to any of embodiments 70-82, wherein composition (a) further comprises one or more preservatives and / or one or more chelating agents and / or one or more antioxidants and / or one or more tonicity adjusters and / or one or more stabilizers and / or one or more viscosity enhancers.

[0295] 84. The composition according to any of embodiments 61-63, wherein the oncolytic peptide is a peptide, peptidomimetic, or amino acid derivative having a net positive charge of at least +2 and comprising disubstituted β-amino acids, each of the substituents in the β-amino acids containing at least 7 non-hydrogen atoms, being lipophilic, and having at least one cyclic group, wherein one or more cyclic groups in a substituent are optionally fused with one or more cyclic groups in another of the substituents.

[0296] 85. The composition according to embodiment 84, wherein one or more cyclic groups in the substituents are fused with one or more cyclic groups in the other substituents, and the combined total number of non-hydrogen atoms for the two substituents is at least 12.

[0297] 86. The oncolytic peptide is a compound comprising a group of formula (IV):

[0298] [ka]

[0299] A composition according to any of embodiments 84-85, wherein X and N have their normal valences and are attached to the rest of the compound; any two of R1, R2, R3, and R4 are hydrogen atoms and two are substituents, which substituents contain at least 7 non-hydrogen atoms, are lipophilic, and contain a cyclic group, which is not directly attached to either the α or β carbon atom and is optionally linked or fused to the cyclic group of another substituent; and X represents O, C, N, or S.

[0300] 87. The composition according to embodiment 86, wherein the substituents are the same or different.

[0301] 88. The composition according to any of embodiments 86-87, wherein the cyclic group is linked or fused to the cyclic group of the other substituent, and the combined total number of non-hydrogen atoms for the two substituents is at least 12.

[0302] 89. The nitrogen atom of the group of formula (VI) is a group R 1~4 and / or the main chain (NC β -C α -CX) five atoms are connected to each other in a linear fashion, and / or R 1~4 is lipophilic in nature, preferably uncharged, preferably has not more than two, more preferably not more than one polar group, and / or X is a substituted N atom.

[0303] 90. The composition according to any of embodiments 86-89, wherein the compound including a group of formula VI has an amidated C-terminus.

[0304] 91. The composition according to any of embodiments 64 to 90, wherein the oncolytic peptide is LTX-401, i.e., a compound of formula (VII).

[0305] [ka]

[0306] 92. The composition according to embodiment 91, wherein LTX-401 is in the form of a pharmaceutically acceptable salt, preferably in the form of a hydrochloride or acetate salt.

[0307] 93. The composition according to any one of embodiments 64 to 92, wherein the oncolytic peptide is present in a concentration of 0.1 to 30 mg / ml, such as 1 to 5 mg / ml or 1 to 30 mg / ml, preferably 4 to 25 mg / ml, or more preferably 6 to 20 mg / ml.

[0308] 94. The composition according to any one of embodiments 64 to 93, wherein the oncolytic peptide is in the form of a pharmaceutically acceptable salt, more preferably in the form of a hydrochloride or acetate salt.

[0309] 95. The composition according to any one of embodiments 64-94, wherein the composition comprises chitosan.

[0310] 96. The composition according to any one of embodiments 64 to 95, wherein the composition comprises several different chitosans.

[0311] 97. The composition according to any one of embodiments 64-96, wherein the chitosan is an ultra-low molecular weight chitosan, a low molecular weight chitosan, and / or an intermediate molecular weight chitosan.

[0312] 98. The composition according to any one of embodiments 64-97, wherein the composition comprises a chitosan derivative.

[0313] 99. The composition according to embodiment 98, wherein the composition comprises several different chitosan derivatives.

[0314] 100. The composition according to any one of embodiments 64 to 99, wherein the chitosan derivative is a glycated chitosan, preferably having a glycation rate of from about 0.1% to about 90% of its otherwise free amino groups and / or having a molecular weight of from about 50 kD to about 2000 kD.

[0315] 101. The composition according to any one of embodiments 64 to 100, wherein the degree of deacetylation (DDA) of the chitosan and / or chitosan derivative is about 50 to 99%, preferably about 70% to about 99%.

[0316] 102. A composition according to any one of embodiments 64 to 101, wherein the composition comprises 0.1 to 5% w / w of chitosan and / or chitosan derivatives, such as 0.1 to 2.5% w / w, such as 0.5%, 1%, 1.5%, or 2% w / w.

[0317] 103. The composition according to any one of embodiments 64-102, wherein the composition is a dispersion, suspension, or solution of i) and ii) in a physiologically acceptable carrier.

[0318] 104. The composition according to any one of embodiments 64 to 103, wherein the pH of the composition is 4.0 to 7.4, more preferably 5.0 to 7.0, for example 6.0 or 6.5.

[0319] 105. The composition according to any one of embodiments 64 to 104, wherein the viscosity of the composition is from 1.0, 1.5, 2, 2.5, or 3 cP to 300 cP, such as 2.5 to 250 cP, such as 5 to 100 cP or 10 to 200 cP and 20 to 240 cP, preferably 40 to 180 cP, such as 45 to 150 cP or 30 to 100 cP, more preferably 1.5 to 10 cP, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 cP, or 10 to 15 cP or 15 to 20 cP or 20 to 35 cP or 35 to 50 cP or 50 to 100 cP.

[0320] 106. The composition according to any one of embodiments 64 to 105, wherein the composition is a pharmaceutical composition.

[0321] 107. A composition according to any one of embodiments 64-106, contained in a sealed container.

[0322] 108. The composition defined in any one of embodiments 64-107, for use as a medicament.

[0323] 109. The composition defined in any one of embodiments 64 to 108, for use in a method for treating a tumor in a subject.

[0324] 110. A composition for use according to embodiment 109, wherein the composition is administered to a subject, for example administered intratumorally.

[0325] 111. A composition for use according to embodiment 110, wherein the composition is administered only once, for example intratumorally.

[0326] 112. A composition for use according to embodiment 110, wherein the composition is administered repeatedly, for example intratumorally.

[0327] 113. A composition for use according to any of embodiments 109 to 112, wherein the composition is administered, for example intratumorally, in a therapeutically effective amount, such amount being administered in one administration or several administrations.

[0328] 114. A composition for use according to any of embodiments 109 to 113, wherein the tumor is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, bone cancer, colorectal cancer, stomach cancer, skin cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, thyroid cancer, kidney cancer, liver cancer, bile duct cancer, brain cancer, cervical cancer, bladder cancer, esophageal cancer, Hodgkin's disease, and adrenocortical carcinoma tumors.

[0329] 115. A composition for use according to any of embodiments 109 to 114, wherein the tumor is a tumor selected from the group consisting of carcinoma, adenocarcinoma, lymphoma, melanoma, blastoma, leukemia, sarcoma, and germ cell tumor.

[0330] 116. A method for preparing a composition as defined in any of embodiments 64 to 106, wherein chitosan and / or chitosan derivatives are dispersed, suspended or dissolved in a physiologically acceptable carrier and an oncolytic peptide is dissolved in said dispersion, suspension or solution, or chitosan and / or chitosan derivatives are dispersed, suspended or dissolved in a solution of oncolytic peptide in a physiologically acceptable carrier.

[0331] 117. The method according to embodiment 116, wherein the composition is sterile filtered and then filled into containers and sealed.

[0332] 118. The method according to embodiment 116, wherein the composition is filled into a container, sealed and autoclaved.

[0333] 119. A kit comprising compositions (a) and (b) as defined in any of embodiments 1-63, or comprising parts / components for preparing said compositions (a) and (b).

[0334] 120. A kit according to embodiment 119, comprising an oncolytic peptide and a physiologically acceptable carrier for preparing composition (a), and chitosan and / or a chitosan derivative and a physiologically acceptable carrier for preparing composition (b), and optionally instructions for preparing compositions (a) and (b).

[0335] 121. The kit according to embodiment 120, wherein said physiologically acceptable carrier for preparing composition (a) is the same as the physiologically acceptable carrier for preparing composition (b).

[0336] 122. A kit according to embodiment 119, comprising composition (a), chitosan and / or a chitosan derivative (e.g., in solid form), and a physiologically acceptable carrier for preparing composition (b), and optionally instructions for preparing composition (b).

[0337] 123. A kit according to embodiment 119, comprising composition (b), an oncolytic peptide (e.g., in solid form), and a physiologically acceptable carrier for preparing composition (a), and optionally instructions for preparing composition (a).

[0338] 124. The kit according to any of embodiments 119-123, further comprising instructions for using compositions (a) and (b).

[0339] 125. A kit according to embodiment 119, comprising compositions (a) and (b) as defined in any of embodiments 1 to 63, and instructions for preparing the composition as defined in any of embodiments 64 to 106 by combining said composition (a) and said composition (b).

[0340] 126. The kit according to embodiment 125, further comprising instructions for using the combination composition.

[0341] 127. A kit comprising parts / components for preparing a composition as defined in any of embodiments 64-106.

[0342] 128. The kit according to embodiment 127, wherein the kit comprises an oncolytic peptide, chitosan and / or a chitosan derivative (e.g., both in solid form), and a physiologically acceptable carrier for preparing a combination composition.

[0343] 129. The kit according to embodiment 128, wherein the oncolytic peptide, chitosan and / or chitosan derivative are contained in one container.

[0344] 130. The kit according to embodiment 128, wherein the oncolytic peptide and the chitosan and / or chitosan derivative are contained in separate containers.

[0345] 131. The kit according to embodiment 127, wherein the kit comprises composition (a) as defined in any of embodiments 1 to 63, and chitosan and / or a chitosan derivative (e.g., in solid form), and optionally instructions for preparing the composition as defined in any of embodiments 64 to 106.

[0346] 132. A kit according to embodiment 127, wherein the kit comprises a composition (b) as defined in any of embodiments 1 to 63, comprising a physiologically acceptable carrier, and an oncolytic peptide (e.g., in solid form), and optionally instructions for preparing the composition as defined in any of embodiments 64 to 106.

[0347] 133. The kit according to any of embodiments 127-132, further comprising instructions for using the composition defined in any of embodiments 64-106.

Claims

1. A composition (a) comprising an oncolytic peptide for use in treating a subject having a tumor by simultaneous, sequential or combined administration with a composition (b) comprising chitosan and / or a chitosan derivative.

2. The composition for use according to claim 1, wherein said composition (a) comprises one or more oncolytic peptides and / or said composition (b) comprises one or more different chitosans and / or chitosan derivatives.

3. The composition for use according to any one of claims 1 to 2, wherein said composition (a) and / or said composition (b) comprises a physiologically acceptable carrier.

4. The composition for use according to any one of claims 1 to 3, wherein the oncolytic peptide is a 9-mer peptide consisting of 9 amino acids in a linear arrangement, or a pharmaceutically acceptable salt thereof, wherein 5 of the 9 amino acids are cationic and 4 are lipophilic, and 3 of the 4 lipophilic amino acids are tryptophan and 1 is a non-genetically encoded amino acid.

5. 5. The composition for use according to claim 4, wherein the cationic amino acid is lysine or arginine and / or the non-genetically encoded amino acid is diphenylalanine or biphenylalanine.

6. The composition for use according to any one of claims 1 to 5, wherein the oncolytic peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329.

7. The composition for use according to any one of claims 1 to 6, wherein the oncolytic peptide is LTX-315.

8. 8. The composition for use according to any of claims 1 to 7, wherein composition (a) further comprises one or more preservatives and / or one or more chelating agents and / or one or more antioxidants and / or one or more tonicity adjusting agents and / or one or more stabilizers and / or one or more viscosity enhancing agents.

9. 4. The composition for use according to any one of claims 1 to 3, wherein the oncolytic peptide is a peptide, peptidomimetic, or amino acid derivative having a net positive charge of at least +2 and comprising disubstituted β-amino acids, each substituent in the β-amino acids containing at least 7 non-hydrogen atoms, being lipophilic, and having at least one cyclic group, wherein one or more cyclic groups in a substituent are optionally fused with one or more cyclic groups in another substituent.

10. The oncolytic peptide is a compound comprising a group of formula (VI): 【Chemistry 1】 where X and N have their normal valences and are bonded to other parts of the compound, and R １ , R ２ , R ３ , and R ４ and X represents O, C, N, or S.

11. 11. The composition for use according to claim 10, wherein said including a group of formula (VI) has an amidated C-terminus.

12. The composition for use according to any of claims 9 to 11, wherein said oncolytic peptide is LTX-401, ie, a compound of formula (VII): 【Chemistry 2】

13. The composition for use according to any one of claims 1 to 12, wherein the oncolytic peptide is in the form of a pharmaceutically acceptable salt.

14. The composition for use according to any one of claims 1 to 13, wherein composition (a) comprises the oncolytic peptide at a concentration of 0.1 to 30 mg / ml.

13. The composition for use according to any one of claims 1 to 12, wherein the pH of composition (a) is from 4.0 to 7.

4.

14. A composition for use according to any one of claims 1 to 13, wherein composition (b) comprises chitosan.

15. The composition for use according to any one of claims 1 to 14, wherein composition (b) comprises a chitosan derivative.

16. The composition for use according to claim 15, wherein the chitosan derivative is a glycated chitosan.

17. The composition for use according to any one of claims 1 to 16, wherein the chitosan and / or chitosan derivative has a degree of deacetylation (DDA) of about 50 to 99%.

18. The composition for use according to any of claims 1 to 17, wherein composition (b) comprises 0.1 to 5% w / w of said chitosan and / or said chitosan derivative.

19. The composition for use according to any one of claims 1 to 18, wherein composition (b) comprises chitosan and / or chitosan derivatives of ultra-low, low or medium molecular weight.

20. 20. The composition for use according to any one of claims 1 to 19, wherein composition (b) is a dispersion, suspension or solution of said chitosan and / or said chitosan derivative in a physiologically acceptable carrier.

21. The composition for use according to any one of claims 1 to 20, wherein the pH of composition (b) is from 4.0 to 7.

5.

22. The composition for use according to any one of claims 1 to 21, wherein the viscosity of composition (a) and / or composition (b) is from 1.0 to 300 cP.

23. 23. The composition for use according to any one of claims 1 to 22, wherein composition (a) is administered into the tumor and composition (b) is administered into the tumor and / or into the tissue immediately surrounding the tumor.

24. 24. The composition for use according to any one of claims 1 to 23, wherein the tumor is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, bone cancer, colorectal cancer, stomach cancer, skin cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, thyroid cancer, kidney cancer, liver cancer, bile duct cancer, brain cancer, cervical cancer, bladder cancer, esophageal cancer, Hodgkin's disease, and adrenocortical carcinoma tumors.

25. A composition comprising i) an oncolytic peptide and ii) chitosan and / or a chitosan derivative.

26. 26. The composition of claim 25, comprising one or more oncolytic peptides and / or one or more different chitosans and / or chitosan derivatives.

27. The composition of any one of claims 25 to 26, wherein the composition comprises a physiologically acceptable carrier.

28. 28. The composition of any of claims 25 to 27, wherein the oncolytic peptide is a 9-mer peptide consisting of nine amino acids in a linear arrangement, or a pharmaceutically acceptable salt thereof, wherein five of the nine amino acids are cationic and four are lipophilic, and three of the four lipophilic amino acids are tryptophan and one is a non-genetically encoded amino acid.

29. 29. The composition of claim 28, wherein the cationic amino acid is lysine or arginine and / or the non-genetically encoded amino acid is diphenylalanine or biphenylalanine.

30. The composition of any of claims 25 to 29, wherein the oncolytic peptide is selected from the group consisting of LTX-302, LTX-313, LTX-315, LTX-320, and LTX-329.

31. The composition according to any one of claims 25 to 30, wherein the oncolytic peptide is LTX-315.

32. 32. The composition of any of claims 25 to 31, further comprising one or more preservatives and / or one or more chelating agents and / or one or more antioxidants and / or one or more tonicity adjusting agents and / or one or more stabilizers and / or one or more viscosity enhancing agents.

33. 28. The composition of any of claims 25 to 27, wherein the oncolytic peptide is a peptide, peptidomimetic, or amino acid derivative having a net positive charge of at least +2 and comprising disubstituted β-amino acids, each substituent in the β-amino acids containing at least 7 non-hydrogen atoms, being lipophilic, and having at least one cyclic group, wherein one or more cyclic groups in a substituent are optionally fused with one or more cyclic groups in another substituent.

34. The oncolytic peptide is a compound comprising a group of formula (VI): 【Transformation 3】 where X and N have their normal valences and are bonded to other parts of the compound, and R １ , R ２ , R ３ , and R ４ and X represents O, C, N, or S.

35. 35. The composition of claim 34, wherein the inclusion of a group of formula (VI) has an amidated C-terminus.

36. The composition according to any one of claims 33 to 35, wherein the oncolytic peptide is LTX-401, i.e., the compound of formula (VII). 【Chemistry 4】

37. The composition of any one of claims 25 to 36, wherein the oncolytic peptide is in the form of a pharmaceutically acceptable salt.

38. The composition according to any one of claims 25 to 37, comprising the oncolytic peptide at a concentration of 0.1 to 30 mg / ml.

39. The composition of any one of claims 25 to 38, comprising chitosan.

40. The composition according to any one of claims 25 to 39, comprising a chitosan derivative.

41. 41. The composition of claim 40, wherein the chitosan derivative is a glycated chitosan.

42. 42. The composition according to any one of claims 25 to 41, wherein the chitosan and / or chitosan derivative has a degree of deacetylation (DDA) of about 50 to 99%.

43. A composition according to any one of claims 25 to 42, comprising 0.1 to 5% w / w of said chitosan and / or said chitosan derivative.

44. 44. The composition according to any one of claims 25 to 43, comprising ultra-low, low or medium molecular weight chitosan and / or chitosan derivatives.

45. 45. The composition of any of claims 25 to 44, wherein the composition is a dispersion, suspension, or solution of i) and ii) in a physiologically acceptable carrier.

46. The composition of any one of claims 25 to 45, wherein the pH of the composition is 4.0 to 7.

4.

47. The composition according to any one of claims 25 to 46, wherein the viscosity of the composition is from 1.0 to 300 cP.

48. A composition according to any one of claims 25 to 47 for use as a medicament.

49. 48. The composition of any of claims 25 to 47 for use in a method for treating a tumor in a subject.

50. A method for preparing the composition of any one of claims 25 to 47, wherein chitosan and / or chitosan derivatives are dispersed, suspended or dissolved in a physiologically acceptable carrier and the oncolytic peptide is dissolved in the dispersion, suspension or solution, or chitosan and / or chitosan derivatives are dispersed, suspended or dissolved in a solution of the oncolytic peptide in the physiologically acceptable carrier.

51. A kit comprising composition (a) and composition (b) according to any one of claims 1 to 24, or comprising parts / components for preparing said compositions (a) and (b).

52. 52. The kit of claim 51, comprising composition (a) and composition (b) of any one of claims 1 to 24, and instructions for preparing the composition of any one of claims 25 to 47 by combining said composition (a) and said composition (b).

53. A kit comprising parts / components for preparing a composition according to any one of claims 25 to 47.

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