Stapled antimicrobial peptides (STAMPs) and their uses

Abstract

Provided are stapled antimicrobial peptides (i.e., StAMPs) and methods of using the same (e.g., for treating bacterial infections caused by gram-negative bacteria). In some embodiments, the stapled peptides are based on the amino acid sequence of the antimicrobial peptide, Magainin II, but include certain modifications that have been found to confer advantageous properties (e.g., improved antimicrobial activity, selectivity for killing gram-negative bacteria, and / or reduced toxicity). Also provided are unstapled peptides that can serve as synthetic precursors to the stapled peptides provided herein.

Description

[Technical field]

[0001] Related Applications This application claims priority under 35 USC § 119(e) to U.S. Provisional Patent Application No. (USSN) 63 / 160,245, filed March 12, 2021, the entire contents of which are incorporated herein by reference.

[0002] Government support This invention was made with Government support under 6 IDSEP160030-04-01 awarded by the Department of Health and Human Services. The Government has certain rights in this invention. [Background technology]

[0003] background The rise of antibiotic resistance worldwide poses a major threat to human health. As the number of newly approved antibiotics has decreased over the past decade, the need for new drugs to combat drug-resistant microorganisms has increased significantly. As the evolution of resistance in bacteria outpaces current drug discovery, there is a need for new drugs to overcome antibiotic resistance, especially in Gram-negative bacteria.

[0004] Antimicrobial peptides (AMPs) are a subclass of naturally occurring peptides expressed by a wide variety of life species to combat infections, primarily of the skin or local compartments. See for example Brogden, KA Nat. Rev. Microbiol. 3, 238-250 (2005); Brown, KL et al. Curr. Opin. Immunol. 18, 24-30 (2006); Hancock, RE et al. Nat. Biotechnol. 24, 1551-1557 (2006). Cationic AMPs kill bacteria by membrane lysis, a mechanism that significantly reduces the likelihood of inducing antibiotic resistance. See for example Fjell, CD et al. Nat. Rev. Drug Discov. 11, 37-51 (2012).

[0005] Indeed, AMPs have been proposed as a potential solution to the global threat of multidrug resistance; however, linear AMPs may be structurally unstable, prone to proteolysis, and / or cause nonspecific membrane toxicity that significantly hinders their clinical translation when administered internally. See, for example, Theuretzbacher, U. Curr. Opin. Microbiol. 39, 106-112 (2017); Woodford, N. et al. J. Antimicrob. Chemother. 63, 225-229 (2009); Blair, JMet al. Nat. Rev. Microbiol. 13, 42-51 (2015). Since their discovery, 30 years of groundbreaking research have characterized natural and non-natural AMPs, but no insight has emerged that would make AMP-based antibiotic drugs clinical candidates. See, for example, Steiner, H. et al. Nature 292, 246-248 (1981).

[0006] One approach to reduce the in vivo proteolytic susceptibility of peptides involves forcing the structure of α-helices by inserting all-hydrocarbon staples. See, for example, Bird, GH et al. Proc. Natl Acad. Sci. USA 107, 14093-14098 (2010); Schafmeister, CE et al. JACS 122, 5891-5892 (2000); Walensky, LD et al. Science 305, 1466-1470 (2004). The application of all-hydrocarbon stapling maximizes the utility of bioactive helices for targeting protein interactions, and the first clinical-grade anticancer stapled peptides are in phase I and II trials. For example, Bird,GHet al. Nat.Chem.Biol.12,845-852(2016); Walensky,LDet al. J.Med.Chem.57,6275-6288(2014); Bird,GHet al. J.Clin.Invest.124,2113-2124(2014); Meric-Bernstam,F.et al. See J. Clin. Oncol. 35, 2505 (2017).

[0007] Attempts to incorporate staples into the AMP sequence have in some cases improved α-helicity and proteolytic resistance, but have resulted in constructs with variable antimicrobial activity and indiscriminate membrane lysis, reflected by a poor therapeutic index. See, for example, Chapuis, H. et. al. Amino Acids 43, 2047-2058 (2012); Dinh, TT et al. Bioorg.Med.Chem.Lett. 25, 4016-4019 (2015); Migon, D. et al. Protein J. 37, 2-12 (2018); Pham, TK et al. Bioorg.Med.Chem.Lett. 23, 6717-6720 (2013); Stone, TA et al. Bioorg.Med.Chem. 26, 1189-1196 (2017); Stone, TA et al. Bioorg.Med.Chem. 26, 1189-1196 (2017). Thus, stapling has been considered as a contributor to nonspecific AMP toxicity in some cases. Summary of the Invention

[0008] SUMMARY OF THE PRESENT APPLICATION Currently, there is a great need for new antimicrobial agents with high antimicrobial activity and low non-specific toxicity. Examples of stapled antimicrobial peptides (StAMPs) and methods of using the same can be found, for example, in International PCT Application Publication No. WO 2017 / 004591, published January 5, 2017; and WO 2019 / 018499, published January 24, 2019, the entire contents of each of which are incorporated herein by reference.

[0009] The present disclosure relates in part to novel stapled antimicrobial peptides (StAMPs) that can be used to treat and / or prevent infectious diseases (e.g., bacterial infections). The stapled peptides provided herein are based on the amino acid sequence of the antimicrobial peptide Magainin II, but include certain modifications that have been found to confer advantageous properties (e.g., improved antimicrobial activity, selectivity for killing gram-negative bacteria, and / or reduced toxicity). The peptides provided herein can be selectively cytotoxic to microbial cells (e.g., bacterial cells) over mammalian cells. In some embodiments, this selectivity translates to reduced toxicity (e.g., reduced nephrotoxicity or hepatotoxicity) in a subject.

[0010] In one aspect, provided herein is a polypeptide having the following amino acid sequence:

number

[0011] For example, in certain embodiments, the stapled peptides provided herein or pharma- ceutically acceptable salts thereof comprise any one of the peptides set forth in SEQ ID NOs: 4-39, 59-77, or 81-100 (below). In certain embodiments, the stapled peptides provided herein or pharma- ceutically acceptable salts thereof comprise any one of the peptides set forth in SEQ ID NOs: 40-53 (below). In certain embodiments, the stapled peptides provided herein or pharma- ceutically acceptable salts thereof comprise any one of the peptides set forth in SEQ ID NOs: 101-105 (below).

[0012] The amino acid sequences and peptides provided herein may contain one or more additional modifications as described herein, such as, for example, amino acid deletions, amino acid additions, C-terminal modifications, and the like.

[0013] In another aspect, provided herein is a pharmaceutical composition comprising the peptide provided herein or its pharmaceutically acceptable salt and one or more pharmaceutically acceptable carriers or excipients.In some embodiments, the pharmaceutical composition provided herein comprises a therapeutically and / or prophylactically effective amount of the peptide provided herein.The pharmaceutical composition described herein is useful for treating and / or preventing disease (e.g., bacterial infection) in a subject.

[0014] In other aspects, provided herein are methods and uses of the compounds and pharmaceutical compositions provided herein, including: (a) A method for treating and / or preventing an infectious disease (e.g., bacterial infection) in a subject, comprising administering to the subject a therapeutically and / or prophylactically effective amount of a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the bacterial infection is caused by a gram-negative bacterium. In some embodiments, the bacterial infection is caused by an antibiotic-resistant bacterium (e.g., an antibiotic-resistant gram-negative bacterium). (b) A method of killing and / or inhibiting the growth of bacteria (e.g., in vivo or in vitro) with a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the bacteria is a gram-negative bacterium. In some embodiments, the bacteria is an antibiotic-resistant gram-negative bacterium. In some embodiments, the bacteria is Escherichia coli (E. coli), Acinetobacter baumannii (A. baumannii), Pseudomonas aeruginosa (P. aeruginosa), or Klebsiella pneumoniae (K. pneumoniae). In some embodiments, the killing and / or inhibition is selective for gram-negative bacteria over gram-positive bacteria. (c) A method for selectively killing and / or inhibiting the growth of microbial cells (e.g., bacterial cells) over mammalian cells, comprising contacting the microbial (e.g., bacterial) cells and mammalian cells with a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. (d) A method for selectively lysing microbial cells (e.g., bacterial cells) over mammalian cells, comprising contacting the microbial (e.g., bacterial) cells and the mammalian cells with a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0015] Also provided herein are peptides described herein (e.g., StAMP), and pharma- ceutically acceptable salts thereof, and pharmaceutical compositions thereof, for use in any of the methods described herein (e.g., for treating an infectious disease in a subject, for treating a bacterial infection in a subject, for killing and / or inhibiting the growth of bacteria in a subject, for selectively killing and / or inhibiting the growth of bacterial cells over mammalian cells in a subject, etc.).

[0016] In another aspect, also provided herein is the use of the peptides described herein (e.g., StAMP), and pharma- ceutically acceptable salts thereof, and pharmaceutical compositions thereof, for the manufacture of a medicament (e.g., for treating an infectious disease in a subject, for treating a bacterial infection in a subject, for killing and / or inhibiting the growth of bacteria in a subject, for selectively killing and / or inhibiting the growth of bacterial cells over mammalian cells in a subject, etc.).

[0017] In another aspect, provided herein is a kit comprising the peptide provided herein (for example, StAMP), or its pharma- ceutically acceptable salt, or its pharmaceutical composition.The kit described herein may comprise a single dose or multiple doses of the peptide or its pharmaceutical composition.The kit described herein is useful in any of the methods or uses provided herein, and optionally further comprises instructions for using the kit (for example, instructions for using the peptide or composition included in the kit).

[0018] In another aspect, provided herein are unstapled or partially stapled peptides that can serve as synthetic precursors to the stapled peptides provided herein. Such unstapled peptides include pairs of amino acids that include reactive moieties capable of forming crosslinks with one another. Also provided herein are methods of preparing the stapled peptides provided herein from the unstapled peptides provided herein. As described herein, unstapled (i.e., uncrosslinked) peptides include pairs of amino acids that include reactive moieties capable of forming crosslinks (e.g., X 1 and X 2 Pairs with and X 3 and X 4 In a preferred embodiment, the amino acid pair (e.g., X 1 and X 2 Pairs with and X 3 and X 4and (pair) comprises an α-side chain containing a reactive moiety. Non-limiting examples of reactive moieties include alkenes, alkynes, alcohols, amines, thiols, azides, esters, amides, halogens, and the like. In some embodiments, two reactive moieties can react directly with each other to form a crosslink (e.g., alkenes via ring-closing metathesis (RCM), or alkynes and azides via 1,3-dipolar cycloaddition to form a triazole). In other embodiments, two reactive moieties react with an intermediate crosslinking reagent to form a crosslink (e.g., two cysteine ​​residues react with a dihalide, e.g., dibromoxylene).

[0019] Details of certain aspects of the invention are set forth in the Detailed Description of Certain Aspects, as set forth below. Other features, objects, and advantages of the invention will be apparent from the definition, examples, figures, and claims.

[0020] definition General definition The following definitions are of general terms used throughout this application.

[0021] The terms "peptide" and "polypeptide" are used interchangeably and refer to a polymer of amino acid residues linked together by peptide bonds. These terms also include proteins and refer to peptides, polypeptides, and proteins of any size, structure, or function. Typically, a peptide will be at least three amino acids long, or at least as long as required by the amino acid sequences provided herein. A peptide may refer to an individual peptide or a group of peptides. The peptides provided herein may include natural amino acids and / or non-natural amino acids (i.e., compounds that do not occur in nature but can be incorporated into a peptide chain) in any combination. One or more of the amino acids in a peptide may be modified, for example, by the addition of a chemical entity (such as a carbohydrate group, a hydroxyl group, a phosphate group, a farnesyl group, an isofarnesyl group, a fatty acid group, a linker for conjugation or functionalization, etc.), or other modification. A peptide may be a fragment or a modified version of a naturally occurring peptide or protein. A peptide may be naturally occurring, recombinant, synthetic, or any combination thereof.

[0022] The peptides provided herein can be of any length. In some embodiments, the peptides are 100 amino acids or less in length. In some embodiments, the peptides are 90 amino acids or less in length. In some embodiments, the peptides are 80 amino acids or less in length. In some embodiments, the peptides are 70 amino acids or less in length. In some embodiments, the peptides are 60 amino acids or less in length. In some embodiments, the peptides are 50 amino acids or less in length. In some embodiments, the peptides are 45 amino acids or less in length. In some embodiments, the peptides are 40 amino acids or less in length. In some embodiments, the peptides are 35 amino acids or less in length. In some embodiments, the peptides are 30 amino acids or less in length. In some embodiments, the peptides are 25 amino acids or less in length. In some embodiments, the peptides are 20 amino acids or less in length. In some embodiments, the peptides are no more than 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 amino acids in length. In some embodiments, the peptides are at least the length of the amino acid sequences provided herein.

[0023] The term "amino acid" refers to a molecule that contains both an amino group and a carboxyl group. Amino acids include alpha-amino acids, the general structure of which is depicted below. Each amino acid referred to herein may also be represented by a one- to four-letter code (for example, R and Arg represent L-arginine, and hArg represents L-homoarginine). [ka]

[0024] Suitable amino acids include, without limitation, natural alpha-amino acids, such as the D- and L-isomers of the 20 common naturally occurring alpha-amino acids found in peptides (e.g., A, R, N, C, D, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y, V, as provided below), and non-naturally occurring alpha-amino acids.

[0025] Exemplary natural alpha-amino acids (single letter codes are provided in parentheses) include L-alanine (A), L-arginine (R), L-asparagine (N), L-aspartic acid (D), L-cysteine ​​(C), L-glutamic acid (E), L-glutamine (Q), glycine (G), L-histidine (H), L-isoleucine (I), L-leucine (L), L-lysine (K), L-methionine (M), L-phenylalanine (F), L-proline (P), L-serine (S), L-threonine (T), L-tryptophan (W), L-tyrosine (Y), and L-valine (V).

[0026] Exemplary unnatural alpha-amino acids are D-arginine, D-asparagine, D-aspartic acid, D-cysteine, D-glutamic acid, D-glutamine, D-histidine, D-isoleucine, D-leucine, D-lysine, D-methionine, D-phenylalanine, D-proline, D-serine, D-threonine, D-tryptophan, D-tyrosine, D-valine, di-vinyl, α-methyl-alanine (Aib), α-methyl-arginine, α-methyl-asparagine ...arginine, α-methyl-asparagine, α-methyl-arginine, α-methyl-asparagine, α-methyl-asparagine, α-methyl-asparagine, α-methyl-asparagine, α-methyl-asparagine, α-methyl-asparagine, α-methyl-asparagine, α-methyl Non-natural amino acids include alpha-amino acids such as spartic acid, alpha-methyl-cysteine, alpha-methyl-glutamic acid, alpha-methyl-glutamine, alpha-methyl-histidine, alpha-methyl-isoleucine, alpha-methyl-leucine, alpha-methyl-lysine, alpha-methyl-methionine, alpha-methyl-phenylalanine, alpha-methyl-proline, alpha-methyl-serine, alpha-methyl-threonine, alpha-methyl-tryptophan, alpha-methyl-tyrosine, alpha-methyl-valine, norleucine, and terminally unsaturated alpha-amino acids. Many non-natural amino acids are known, any of which may be included in the peptides of the present disclosure. See, for example, S. Hunt, The Non-Protein Amino Acids: In Chemistry and Biochemistry of the Amino Acids, edited by GC Barrett, Chapman and Hall, 1985. Non-natural amino acids also include amino acids that contain nitrogenous substituents (e.g., non-hydrogen groups).

[0027] Certain amino acids referred to herein are provided below in Table 1 (represented by name, structure, and one- to four-letter code). Table 1. Certain amino acids [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0028] The term "amino acid substitution", when used in reference to an amino acid sequence, refers to an amino acid of an amino acid sequence being replaced by a different amino acid (e.g., replaced by a natural or non-natural amino acid). The amino acid sequences provided herein may contain or include one or more amino acid substitutions. A particular amino acid substitution is indicated by a colloquial nomenclature commonly used in the art of peptide sequencing to indicate variations in amino acid sequences. For example, with reference to SEQ ID NO:2 (below), "amino acid substitution at H7" refers to a histidine (H) at position 7 of the amino acid sequence being replaced by a different amino acid (e.g., a natural or non-natural amino acid other than histidine). Also with reference to SEQ ID NO:2, the amino acid substitution "H7K" refers to the replacement of the histidine (H) at position 7 of the amino acid sequence represented by SEQ ID NO:2 with a lysine (K), resulting in the amino acid sequence represented by SEQ ID NO:4 (below).

number

[0029] The term "amino acid addition", when used in reference to an amino acid sequence, refers to an amino acid (e.g., a natural or unnatural amino acid) being inserted between two amino acids of the amino acid sequence or added at either end of the sequence. Standard colloquial nomenclature is used to represent the specific amino acid addition (e.g., with reference to SEQ ID NO:2, "G3_K4insX" indicates that a hypothetical amino acid X is inserted between amino acids G3 and K4 of the amino acid sequence). In some embodiments, the amino acid sequence herein may include 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid additions.

[0030] The term "amino acid deletion", when used in reference to an amino acid sequence, refers to an amino acid of the amino acid sequence being deleted from the amino acid sequence. Standard colloquial nomenclature is used to indicate the specific amino acid deletion (for example, with reference to SEQ ID NO:2, "G1del" indicates that the amino acid G1 is deleted from the sequence). In some embodiments, the amino acid sequence herein can include 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid deletions.

[0031] The terms "stapled" and "bridged" are used interchangeably and refer to a peptide in which two amino acids (i.e., the bridged amino acids) are connected (i.e., stapled) via a bridge to form a macrocycle. The terms "bridge" and "staple" are used interchangeably and refer to a covalent linking moiety, other than the peptide backbone, that connects the bridged amino acid pair to form the macrocycle. In some embodiments, the bridge is attached to the alpha position of the bridged amino acid. In some embodiments, the bridged amino acids have three amino acids between them in the amino acid sequence, forming an "i+4 bridge". In some embodiments, the bridged amino acids have four amino acids between them in the amino acid sequence, forming an "i+5 bridge". In some embodiments, the bridged amino acids have six amino acids between them in the amino acid sequence, forming an "i+7 bridge". In some embodiments, the bridged amino acids have seven amino acids between them in the amino acid sequence, forming an "i+8 bridge".

[0032] For example, the following structure has the amino acid sequence GX 1 GGGX 2 (SEQ ID NO: 54), where X 1 and X 2 are amino acids connected via an i+4 bridge of formula (alk), the peptide comprising a C-terminus amidated with -NH2: [ka]

[0033] In some embodiments, the stapled peptides provided herein are based on the amino acid sequence of the antimicrobial peptide Magainin II. Magainin is a class of antimicrobial peptides (AMPs) that were first discovered in Xenopus laevis. The peptides are cationic and generally lack a stable conformation in water, but form amphipathic α-helices in membranes. They are generally known to disrupt the cell membranes of a wide range of cells, including bacteria, protozoa, and fungi. The amino acid sequences of the peptides known as "Magainin I" and "Magainin II" are provided below.

number

[0034] As used herein, "stapleted antimicrobial peptide" (ie, "StAMP") refers to a stapled peptide that has antimicrobial (eg, antibacterial) activity.

[0035] As used herein, the term "salt" refers to any and all salts and encompasses pharma- ceutically acceptable salts. Salts include ionic compounds resulting from the neutralization reaction of acids and bases. Salts are composed of one or more cations (positively charged ions) and one or more anions (negative ions) so that the salt is electrically neutral (no net charge). Salts of the peptides of the present invention include salts derived from inorganic and organic acids and bases. Examples of acid addition salts are salts of amino groups formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid) or by using other methods known in the art, such as ion exchange. Other salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, dicurconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobiotin, and tetrahydrofuran. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N-type salts such as phosphate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, hippurate, and the like. + (C 1~4Representative salts of alkali or alkaline earth metals include sodium, lithium, potassium, calcium, magnesium, etc. Further salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0036] The term "pharmaceutically acceptable salt" refers to a salt that is suitable for use in contact with the tissues of humans and lower animals, within the bounds of sound medical wisdom, without excessive toxicity, irritation, allergic responses, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19 (incorporated herein by reference). Pharmaceutically acceptable salts of the peptides of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid), or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or by using other methods known in the art (such as ion exchange). Other pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N-type salts. + (C 1~4 Alkyl)4 -Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharma-ceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates, and arylsulfonates.

[0037] Throughout this disclosure, references to "the peptide" and "a peptide" as provided herein are intended to encompass peptides comprising any of the amino acid sequences provided herein (including any disclosed amino acid substitutions, additions, deletions, and / or modifications), as well as pharma- ceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, as described herein.

[0038] The terms "composition" and "formulation" are used interchangeably.

[0039] A "subject" to which administration is contemplated refers to a human (i.e., male or female of any age group, e.g., a pediatric subject (e.g., an infant, a child, a juvenile), or an adult subject (e.g., a young adult, a middle-aged adult, or an elderly adult)), or a non-human animal. In some embodiments, the non-human animal is a mammal (e.g., a primate (e.g., a cynomolgus monkey or a rhesus monkey), a commercially relevant mammal (e.g., a cattle, a pig, a horse, a sheep, a goat, a cat, or a dog), or a bird (e.g., a commercially relevant bird, such as a chicken, a duck, a goose, or a turkey)). In some embodiments, the non-human animal is a fish, a reptile, or an amphibian. The non-human animal may be male or female at any stage of development. The non-human animal may be a transgenic or genetically modified animal. The term "patient" refers to a human subject in need of treatment for a disease.

[0040] The term "biological sample" refers to any sample, including tissue samples (such as tissue sections and tissue needle biopsies); cell samples (e.g., cytological smears (such as Pap or blood smears) or samples obtained by microdissection); whole organism samples (such as yeast or bacterial samples); or cell fractions, fragments, or organelles (such as those obtained by lysing cells and centrifuging or otherwise separating their components). Other examples of biological samples include blood, serum, urine, semen, feces, cerebrospinal fluid, interstitial fluid, mucus, tears, sweat, pus, biopsy tissue (e.g., obtained by surgical or needle biopsy), nipple aspirate, milk, vaginal fluid, saliva, swabs (such as buccal swabs), or any material containing biological molecules derived from an initial biological sample.

[0041] The terms "administer," "administering," or "administration" refer to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a peptide described herein or a composition thereof into or onto a subject.

[0042] The terms "treatment", "treat" and "treating" refer to arresting, alleviating, delaying the onset or inhibiting the progression of a disease as described herein. In some embodiments, treatment may be administered after one or more signs or symptoms of a disease have developed or been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of a disease or illness. For example, treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., in light of a history of symptoms and / or exposure to a pathogen). Treatment may also be continued after symptoms have resolved, for example, to delay and / or prevent recurrence.

[0043] The terms "disease," "disease," and "disorder" are used interchangeably.

[0044] An "effective amount" of a peptide described herein refers to an amount sufficient to elicit a desired biological response. An effective amount of a peptide described herein may vary depending on factors such as the desired biological endpoint, side effects, severity of the disease or disorder, identity of the specific peptide, pharmacokinetics and pharmacodynamics, condition being treated, mode of administration, route of administration, and desired or required frequency of administration, species, age, and health or general condition of the subject. In some embodiments, an effective amount is a therapeutically effective amount. In some embodiments, an effective amount is a prophylactic treatment. In some embodiments, an effective amount is the amount of a peptide described herein in a single dose. In some embodiments, an effective amount is the combined amount of a peptide described herein in multiple doses.

[0045] A "therapeutically effective amount" of a peptide as described herein is an amount sufficient to provide a therapeutic benefit in treating a disease or to delay or minimize one or more symptoms associated with a disease. A therapeutically effective amount of a peptide refers to an amount of a therapeutic agent alone, or in combination with other therapies, that provides a therapeutic benefit in treating a disease. The term "therapeutically effective amount" can encompass an amount that improves overall treatment, an amount that reduces or avoids symptoms, signs, or causes of a disease, and / or an amount that enhances the therapeutic effectiveness of another therapeutic agent.

[0046] A "prophylactically effective amount" of a peptide as described herein is an amount sufficient to prevent or prevent the recurrence of a disease or one or more symptoms associated with a disease. A prophylactically effective amount of a peptide refers to an amount of therapeutic agent alone, or in combination with other agents, that provides a prophylactic benefit in terms of preventing a disease. The term "prophylactically effective amount" can encompass an amount that improves overall prophylaxis or an amount that enhances the prophylactic effectiveness of another prophylactic agent.

[0047] The terms "prevent", "preventing", or "prevention" refer to prophylactic treatment of a subject who does not have the disease and has never had the disease but is at risk of developing the disease, or a subject who has had the disease and is now at risk of developing a relapse of the disease but does not have the disease. In some embodiments, the subject is at higher risk of developing the disease or at higher risk of developing a relapse of the disease than the average member of a healthy population.

[0048] "Infectious disease" refers to any disease caused by a pathogen (i.e., a pathogenic microorganism). An infectious disease may be caused by a bacterium, a virus, a parasite, or a fungus. An infectious disease may be a microbial infection. "Microbial infection" refers to an infection caused by a microorganism, such as a fungus, a bacterium, or a virus. Various bacterial infections include, but are not limited to, skin infections (e.g., bacterial cellulitis, wound infections), gastrointestinal infections, throat infections (e.g., strep throat), urinary tract infections (UTIs), urogenital infections, sexually transmitted diseases (e.g., gonorrhea, chlamydia, syphilis), lung infections (e.g., pneumonia, pneumococcal pneumonia, tuberculosis, whooping cough), food poisoning, sepsis, bacterial meningitis, Lyme disease, cholera, botulism, tetanus, anthrax, blood infections, and systemic infections. In some embodiments, the microbial infection is an infection caused by a bacterium, i.e., a "bacterial infection." In some embodiments, the bacteria is a gram-negative bacteria. In some embodiments, the bacteria is a gram-positive bacteria.

[0049] "Gram-negative bacteria" were originally defined by their ability not to retain the Gram stain; however, since then, Gram-negative bacteria have generally been further defined as bacteria having a cell wall with a thin peptidoglycan layer, and having a lipid outer membrane.

[0050] In some embodiments, the gram-negative bacteria is an Escherichia species. In some embodiments, the gram-negative bacteria is an Escherichia coli (E. coli) strain. In some embodiments, the gram-negative bacteria is a strain of Escherichia albertii, Escherichia blattae, Escherichia fergusonii, Escherichia hermannii, or Escherichia vulneris.

[0051] In some embodiments, the gram-negative bacteria is a Pseudomonas species. In some embodiments, the gram-negative bacteria is a Pseudomonas aeruginosa strain. In some embodiments, the gram-negative bacteria is a strain of Pseudomonas alcaligenes, Pseudomonas anguilliseptica, Pseudomonas argentinensis, Pseudomonas borbori, Pseudomonas citronellolis, Pseudomonas flavescens, Pseudomonas mendocina, Pseudomonas nitroreducens, Pseudomonas oleovorans, Pseudomonas pseudoalcaligenes, Pseudomonas resinovorans, Pseudomonas straminea, Pseudomonas chlororaphis, Pseudomonas fluorescens, Pseudomonas pertucinogena, Pseudomonas putida, Pseudomonas stutzeri, or Pseudomonas syringae.

[0052] In some embodiments, the gram-negative bacterium is a Klebsiella species. In some embodiments, the gram-negative bacterium is a strain of Klebsiella granulomatis, Klebsiella oxytoca, Klebsiella pneumoniae, Klebsiella terrigena, or Klebsiella planticola.

[0053] In some embodiments, the gram-negative bacterium is a Salmonella species. In some embodiments, the gram-negative bacterium is a Salmonella bongori strain or a Salmonella enterica strain, such as Salmonella typhi.

[0054] In some embodiments, the gram-negative bacterium is an Acinetobacter species. In some embodiments, the gram-negative bacterium is an Acinetobacter baumannii strain. In certain embodiments, the gram-negative bacterium is a strain of Acinetobacter baylyi, Acinetobacter bouvetii, Acinetobacter calcoaceticus, Acinetobacter gerneri, Acinetobacter grimontii, Acinetobacter haemolyticus, Acinetobacter johnsonii, Acinetobacter junii, Acinetobacter lwoffii, Acinetobacter parvus, Acinetobacter pittii, Acinetobacter radioresistens, Acinetobacter schindleri, Acinetobacter tandoii, Acinetobacter tjernbergiae, Acinetobacter towneri, Acinetobacter ursingii, or Acinetobacter gyllenbergii.

[0055] In some embodiments, the gram-negative bacteria is a Stenotrophomonas species. In some embodiments, the gram-negative bacteria is a strain of Stenotrophomonas acidaminiphila, Stenotrophomonas bentonitica, Stenotrophomonas chelatiphaga, Stenotrophomonas daejeonensis, Stenotrophomonas dokdonensis, Stenotrophomonas ginsengisoli, Stenotrophomonas humi, Stenotrophomonas indicatrix, Stenotrophomonas koreensis, Stenotrophomonas lactitubi, Stenotrophomonas maltophilia, Stenotrophomonas nitritireducens, Stenotrophomonas pavanii, Stenotrophomonas pictorum, Stenotrophomonas rhizophila, Stenotrophomonas terrae, or Stenotrophomonas tumulicola.

[0056] In some embodiments, the gram-negative bacterium is a Burkholderia species. In some embodiments, the bacterium is a strain of Burkholderia cepacian, Burkholderia multivorans, Burkholderia cenocepacia, Burkholderia vietnamiensis, Burkholderia stabilis, Burkholderia ambifaria, Burkholderia dolosa, Burkholderia anthina, Burkholderia pyrrocinia, or Burkholderia ubonensis.

[0057] In some embodiments, the gram-negative bacterium is a Neisseria species, hi some embodiments, the bacterium is a strain of Neisseria meningitidis or Neisseria gonorrhoeae.

[0058] In some embodiments, the gram-negative bacterium is an Enterobacter species. In some embodiments, the bacterium is a strain of Enterobacter amnigenus, Enterobacter arachidis, Enterobacter asburiae, Enterobacter cancerogenous, Enterobacter cloacae, Enterobacter cowanii, Enterobacter dissolvens, Enterobacter gergoviae, Enterobacter helveticus, Enterobacter hormaechei, Enterobacter intermedius, Enterobacter kobei, Enterobacter ludwigii, Enterobacter mori, Enterobacter nimipressuralis, Enterobacter oryzae, Enterobacter pulveris, Enterobacter pyrinus, Enterobacter radicincitans, Enterobacter taylorae, Enterobacter turicensis, or Enterobacter soli.

[0059] In certain embodiments, the Gram-negative bacterium is a Pasteurella sp. In certain embodiments, the bacteria are Pasteurella aerogenes, Pasteurella anatis, Pasteurella avium, Pasteurella bettyae, Pasteurella caballi, Pasteurella canis, Pasteurella dagmatis, Pasteurella gallicida, Pasteurella gallinarum, Pasteurella granulomatis, Pasteurella langaaensis, Pasteurella lymphangitidis, Pasteurella mairii, Pasteurella multocida, Pasteurella oralis, Pasteurella pneumotropica, Pasteurella skyensis, Pasteurella stomatis, Pasteurella testudinis, Pasteurella trehalosi, Pasteurella ureae, or Pasteurella volantium.

[0060] In some embodiments, the gram-negative bacterium is a Proteus species, hi some embodiments, the bacterium is a strain of Proteus hauseri, Proteus mirabilis, Proteus myxofaciens, Proteus penneri, or Proteus vulgaris.

[0061] In some embodiments, the gram-negative bacterium is a Citrobacter species. In some embodiments, the bacterium is a strain of Citrobacter amalonaticus, Citrobacter braakii, Citrobacter diversus, Citrobacter europaeus, Citrobacter farmer, Citrobacter freundii, Citrobacter gillenii, Citrobacter koseri, Citrobacter murliniae, Citrobacter pasteurii, Citrobacter rodentium, Citrobacter sedlakii, Citrobacter werkmanii, Citrobacter youngae, or Citrobacter portucalensis.

[0062] In one embodiment, the gram-negative bacteria are Helicobacter species. In one embodiment, the bacteria are Helicobacter acinonychis, Helicobacter ailurogastricus, Helicobacter anseris, Helicobacter aurati, Helicobacter apri, Helicobacter baculiformis, Helicobacter bilis, Helicobacter bizzozeronii, Helicobacter brantae, Helicobacter canadensis, Helicobacter canicola, Helicobacter canis, Helicobacter cetorum, Helicobacter cholecystus, Helicobacter cinaedi, Helicobacter cynogastricus, Helicobacter equorum, Helicobacter felis, Helicobacter fennelliae, Helicobacter ganmani, Helicobacter heilmannii, Helicobacter hepaticus, Helicobacter himalayensis, Helicobacter japonicus, Helicobacter mesocricetorum, Helicobacter macacae, Helicobacter marmotae, Helicobacter mastomyrinus, Helicobacter mesocricetorum, Helicobacter muridarum, Helicobacter mustelae, Helicobacter pametensis, Helicobacter pullorum, Helicobacter pylori, Helicobacter rappini, Helicobacter rodentium, Helicobacter saguini, Helicobacter salomonis, Helicobacter suis, Helicobacter trogontum, Helicobacter typhlonius, Helicobactervaldiviensis, or a strain of Helicobacter winghamensis.

[0063] In some embodiments, the gram-negative bacterium is a Campylobacter species. In certain embodiments, the bacteria are Campylobacter avium, Campylobacter butzleri, Campylobacter canadensis, Campylobacter cinaedi, Campylobacter coli, Campylobacter concisus, Campylobacter corcagiensis, Campylobacter cryaerophilus, Campylobacter cuniculorum, Campylobacter curvus, Campylobacter fennelliae, Campylobacter fetus, Campylobacter gracilis, Campylobacter helveticus, Campylobacter hepaticus, Campylobacter hominis, Campylobacter hyoilei, Campylobacter hyointestinalis, Campylobacter insulaenigrae, Campylobacter jejuni, Campylobacter lanienae, Campylobacter lari, Campylobacter mucosalis, Campylobacter mustelae, Campylobacter nitrofigilis, Campylobacter peloridis, Campylobacter pylori, Campylobacter rectus, Campylobacter showae, Campylobacter sputorum, Campylobacter subantarcticus, Campylobacter upsaliensis, Campylobacter ureolyticus, or a strain of Campylobacter volucris.

[0064] "Gram-positive bacteria" are those that take up crystal violet dye in the Gram staining test. ,in general They are bacteria that have a cell wall that contains a thin peptidoglycan layer but no lipid outer membrane.

[0065] In some embodiments, the gram-positive bacteria is a Staphylococcus species. In some embodiments, the bacteria is Staphylococcus aureus (S. aureus). In some embodiments, the bacteria is methicillin-resistant Staphylococcus aureus (MRSA). In some embodiments, the bacteria is a Staphylococcus epidermidis (S. epidermidis) strain. In certain embodiments, the bacteria are Staphylococcus auricularis, Staphylococcus carnosus, Staphylococcus condimenti, Staphylococcus massiliensis, Staphylococcus piscifermentans, Staphylococcus simulans, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus saccharolyticus, Staphylococcus devriesei, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus chromogenes, Staphylococcus felis, Staphylococcus delphini, Staphylococcus hyicus, Staphylococcus intermedius, Staphylococcus lutrae, Staphylococcus microti, Staphylococcus muscae, Staphylococcus pseudintermedius, Staphylococcus rostri, Staphylococcus schleiferi, Staphylococcus lugdunensis, Staphylococcus arlettae, Staphylococcus cohnii, Staphylococcus equorum, Staphylococcus gallinarum, Staphylococcus kloosii, Staphylococcus leei, Staphylococcus nepalensis, Staphylococcussaprophyticus, Staphylococcus succinus, Staphylococcus xylosus, Staphylococcus fleurettii, Staphylococcus lentus, Staphylococcus sciuri, Staphylococcus stepanovicii, Staphylococcus vitulinus, Staphylococcus simulans, Staphylococcus pasteuri, or Staphylococcus warneri strains.

[0066] In certain embodiments, the Gram-positive bacterium is of the genus Streptococcus. In certain embodiments, the bacterium is a strain of Streptococcus agalactiae, Streptococcus anginosus, Streptococcus bovis, Streptococcus canis, Streptococcus constellatus, Streptococcus dysgalactiae, Streptococcus equinus, Streptococcus iniae, Streptococcus intermedius, Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, Streptococcus parasanguinis, Streptococcus peroris, Streptococcus pneumoniae, Streptococcus pseudopneumoniae, Streptococcus pyogenes, Streptococcus ratti, Streptococcus salivarius, Streptococcus tigurinus, Streptococcus thermophilus, Streptococcus sanguinis, Streptococcus sobrinus, Streptococcus suis, Streptococcus uberis, Streptococcus vestibularis, Streptococcus viridans, or Streptococcus zooepidemicus.

[0067] In certain embodiments, the Gram-positive bacterium is of the genus Enterococcus. In certain embodiments, the bacterium is a strain of Enterococcus avium, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae, or Enterococcus solitarius.

[0068] In some embodiments, the gram-positive bacterium is a Listeria species. In some embodiments, the bacterium is a strain of Listeria fleischmannii, Listeria grayi, Listeria innocua, Listeria ivanovii, Listeria marthii, Listeria monocytogenes, Listeria rocourtiae, Listeria seeligeri, Listeria weihenstephanensis, or Listeria welshimeri.

[0069] In one embodiment, the gram-positive bacteria are Clostridium species. In one embodiment, the bacteria are Clostridium acetobutylicum, Clostridium argentinense, Clostridium aerotolerans, Clostridium baratii, Clostridium beijerinckii, Clostridium bifermentans, Clostridium botulinum, Clostridium butyricum, Clostridium cadaveris, Clostridium cellulolyticum, Clostridium chauvoei, Clostridium clostridioforme, Clostridium colicanis, Clostridium difficile, Clostridium estertheticum, Clostridium fallax, Clostridium feseri, Clostridium formicaceticum, Clostridium histolyticum, Clostridium innocuum, Clostridium kluyveri, Clostridium ljungdahlii, Clostridium lavalense, Clostridium leptum, Clostridium novyi, Clostridium oedematiens, Clostridium paraputrificum, Clostridium perfringens (also known as Clostridium welchii), Clostridium phytofermentans, Clostridium piliforme, Clostridium ragsdalei, Clostridium ramosum, Clostridium scatologenes, Clostridium septicum, Clostridium sordellii, Clostridium sporogenes, Clostridium sticklandii, Clostridium tertium, Clostridium tetani, ClostridiumThe most common strains of Clostridium thermocellum, Clostridium thermosaccharolyticum, or Clostridium tyrobutyricum.

[0070] As used herein, an "antibiotic-resistant bacterial infection" is a bacterial infection caused by an antibiotic-resistant bacterium. "Antibiotic resistance" occurs when a microorganism evolves a mechanism to protect itself from the effects of antibiotics. A microorganism that is resistant to multiple antimicrobial drugs is called "multidrug-resistant" (MDR). In some embodiments, the methods herein are for treating a multidrug-resistant bacterial infection. In some embodiments, the methods herein are for killing and / or inhibiting the growth of a multidrug-resistant bacterial infection.

[0071] As used herein, the term "inhibit" or "inhibition" in the context of bacterial growth refers to, for example, a reduction in the rate of bacterial growth. In some embodiments, the term refers to a reduction in the rate of bacterial growth to a level that is statistically significantly slower than the initial rate (e.g., the bacterial growth rate before administration or application of the antimicrobial peptides provided herein). In some embodiments, the term refers to a reduction in the rate of bacterial growth to a rate that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of the initial rate (e.g., the bacterial growth rate before administration or application of the antimicrobial peptides provided herein).

[0072] A peptide described herein "selectively" kills and / or "selectively" inhibits the growth of one type of bacteria over another if it kills and / or inhibits the growth of one type of bacteria to a greater extent than the other (e.g., selectively kills and / or selectively inhibits the growth of gram-negative bacteria over gram-positive bacteria). A peptide described herein "selectively" kills and / or "selectively" inhibits the growth of microbial cells over mammalian cells if it kills and / or inhibits the growth of microbial cells to a greater extent than mammalian cells (e.g., selectively kills and / or selectively inhibits the growth of bacterial cells over mammalian cells). A peptide described herein "selectively" lyses microbial cells over mammalian cells if it lyses microbial cells to a greater extent than mammalian cells (e.g., selectively lyses bacterial cells over mammalian cells). A peptide described herein is "selectively" cytotoxic to microbial cells (e.g., bacterial cells) over mammalian cells if it is significantly more toxic (e.g., by lysing, killing or otherwise damaging) to microbial cells (e.g., bacterial cells) than mammalian cells. In some embodiments, the selectivity in any of the above embodiments is at least 1.1-fold, at least 1.5-fold, 2-fold, at least 3-fold, at least 5-fold, at least 10-fold, at least 30-fold, at least 50-fold, at least 100-fold, at least 300-fold, at least 500-fold, at least 1,000-fold, at least 3,000-fold, at least 5,000-fold, at least 10,000-fold, at least 30,000-fold, at least 50,000-fold, or at least 100,000-fold. In some embodiments, the selectivity is 100,000-fold or less, 10,000-fold or less, 100-fold or less, 10-fold or less, or 2-fold or less. Combinations of the above ranges (e.g., at least 2-fold and 10,000-fold or less) are also within the scope of the present disclosure.

[0073] Chemical Definition Definitions of specific functional groups and chemical terms are described in more detail below. Elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as set forth therein. In addition, general principles of organic chemistry and specific functional moieties and reactivity are described in detail in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Michael B. Smith, March's Advanced Organic Chemistry, 7 th Edition, John Wiley & Sons, Inc., New York, 2013; Richard C. Larock, Comprehensive Organic Transformations, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

[0074] The peptides described herein may contain one or more asymmetric centers, and therefore may exist in various stereoisomeric forms, for example, enantiomers and / or diastereomers.For example, the peptides described herein may exist in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers.Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, ELStereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH, Table of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The present invention additionally encompasses peptides as individual isomers substantially free of other isomers, or alternatively as mixtures of various isomers.

[0075] In the formula, the bond [ka] is a single bond, and the dashed line [ka] is a single bond or is absent, [ka] is a single bond or a double bond. [ka] is a double bond or a triple bond.

[0076] Unless otherwise specified, the formulas and structures depicted herein include peptides that do not include isotopically enriched atoms, and also include peptides that include isotopically enriched atoms ("isotopically labeled derivatives"), e.g., hydrogen has been replaced by deuterium or tritium, or 19 F 18 F or carbon is replaced 13 C or 14 Peptides having this structure, except that C is replaced by enriched carbon, are also within the scope of this disclosure. Such peptides are useful, for example, as analytical tools or probes in biological assays. The term "isotopes" refers to variants of a particular chemical element such that all isotopes of a given element share the same number of protons in each atom of the element, but the isotopes differ in the number of neutrons.

[0077] When a range of values ​​("range") is listed, each value and subrange within the range is encompassed. The range also includes the two endpoints of the range unless otherwise specified. For example, "C 1~6 Alkyl" is C1, C2, C3, C4, C5, C6, C 1~6 , C 1~5 , C 1~4 , C 1~3 , C 1~2 , C 2~6 , C 2~5 , C 2~4 , C 2~3 , C 3~6 , C 3~5 , C 3~4 , C 4~6 , C 4~5 , and C 5~6 Covers alkyl.

[0078] Use of the phrase "at least one" refers to 1, 2, 3, 4, or more, but also covers ranges, examples such as from 1 to 4, from 1 to 3, from 1 to 2, from 2 to 4, from 2 to 3, or from 3 to 4 (inclusive).

[0079] A "non-hydrogen group" refers to any group defined for a specific variable that is not hydrogen.

[0080] The term "aliphatic" refers to alkyl, alkenyl, alkynyl, and carbocyclic groups, as well as heteroalkyl, heteroalkenyl, heteroalkynyl, and heterocyclic groups.

[0081] The term "alkyl" refers to the radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms ("C 1~20 In some embodiments, an alkyl group has 1 to 6 carbon atoms ("C 1~6 "Alkyl"). C 1~6 Examples of alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tert-amyl), and hexyl (C6) (e.g., n-hexyl). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8), n-dodecyl (C9), n-hexyl (C10), n-hexyl (C11), n-hexyl (C12), n-hexyl (C13), n-hexyl (C14), n-hexyl (C15), n-hexyl (C16), n-hexyl (C17), n-hexyl (C18), n-hexyl (C19), n-hexyl (C20), n-hexyl (C210), n-hexyl (C220), n-hexyl (C23), n-hexyl (C24), n-hexyl (C25), n-hexyl (C26), n-hexyl (C27), n-hexyl (C28), n-hexyl (C29), n-hexyl (C30), n-hexyl (C31), n-hexyl (C32), n-hexyl (C33), n-hexyl (C34), n-hexyl (C35), n-hexyl (C36), n-hexyl (C37), n-hexyl (C38), n-hexyl (C39), n-hexyl (C31), n-hexyl (C32), n-hexyl (C33), n-hexyl (C34), n-hexyl (C35), n-hexyl (C36), n-hexyl (C37), n-hexyl (C 12 ), etc.

[0082] The term "haloalkyl" refers to a substituted alkyl group in which one or more of the hydrogen atoms are independently replaced by a halogen, such as fluoro, bromo, chloro, or iodo. "Perhaloalkyl" is a subset of haloalkyl and refers to an alkyl group in which all of the hydrogen atoms are independently replaced by a halogen, such as fluoro, bromo, chloro, or iodo. In some embodiments, the haloalkyl moiety has 1 to 20 carbon atoms ("C 1~20 In some embodiments, all of the haloalkyl hydrogen atoms are independently replaced with fluoro to provide a "perfluoroalkyl" group. In some embodiments, all of the haloalkyl hydrogen atoms are independently replaced with chloro to provide a "perchloroalkyl" group. Examples of haloalkyl groups include -CHF2, -CH2F, -CF3, -CH2CF3, -CF2CF3, -CF2CF2CF3, -CCl3, -CFCl2, -CF2Cl, and the like.

[0083] The term "heteroalkyl" refers to an alkyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, located within the parent chain (e.g., inserted between adjacent carbon atoms) and / or at one or more terminal position(s). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~20 "alkyl").

[0084] The term "alkenyl" refers to the radical of a straight-chain or branched hydrocarbon group having from 1 to 20 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds). In some embodiments, an alkenyl group has 1-20 carbon atoms ("C 1~20The one or more carbon-carbon double bonds can be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). In an alkenyl group, a C=C double bond of unspecified stereochemistry (e.g., -CH=CHCH3 or [ka] may be in the (E)- or (Z)-configuration.

[0085] The term "heteroalkenyl" refers to an alkenyl further including at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, located within the parent chain (e.g., inserted between adjacent carbons) and / or at one or more terminal position(s). In certain embodiments, a heteroalkenyl group refers to a group having from 1 to 20 carbon atoms, at least one double bond, and one or more heteroatoms in the parent chain ("heteroC 1~20 alkenyl).

[0086] The term "alkynyl" refers to a straight-chain or branched hydrocarbon radical having from 1 to 20 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) (see "C 1~20 The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).

[0087] The term "heteroalkynyl" refers to an alkynyl group further including at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, located within the parent chain (e.g., inserted between adjacent carbons) and / or at one or more terminal position(s). In certain embodiments, a heteroalkynyl group refers to a group having from 1 to 20 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroalkynyl" refers to a group having from 1 to 20 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroalkynyl" refers to a group having from 1 to 20 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroalkynyl" refers to a group having from 1 to 20 carbon atoms, at least one triple bond, and at least one heteroatom ...). 1~20alkynyl").

[0088] The term "carbocyclyl" or "carbocyclic" refers to a ring system having from 3 to 14 ring carbon atoms ("C 3~14 In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C 3~6 Carbocyclyl). 3~6 Carbocyclyl groups include cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), etc. As the examples above illustrate, in certain embodiments, carbocyclyl groups are either monocyclic ("monocyclic carbocyclyl") or polycyclic (containing, for example, fused, bridged, or spiro ring systems, such as a bicyclic ("bicyclic carbocyclyl") or tricyclic ("tricyclic carbocyclyl") system), and may be saturated or may contain one or more carbon-carbon double or triple bonds. "Carbocyclyl" also encompasses ring systems in which a carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups and the point of attachment is on the carbocyclyl ring, in which case the number of carbons is designated uninterrupted as the number of carbons in the carbocyclyl ring system.

[0089] The term "heterocyclyl" or "heterocyclic" refers to a radical of a 3- to 14-membered non-aromatic ring system having ring carbon atoms and one to four ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("3- to 14-membered heterocyclyl"). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be at a carbon atom or at a nitrogen atom, where valence permits. In some embodiments, the heterocyclyl is a substituted or unsubstituted 3- to 7-membered monocyclic heterocyclyl, where one, two, or three atoms in the heterocyclic ring system are independently oxygen, nitrogen, or sulfur, where valence permits. Heterocyclyl groups can be either monocyclic ("monocyclic heterocyclyl") or polycyclic (e.g., fused, bridged, or spiro ring systems, such as bicyclic ("bicyclic heterocyclyl") or tricyclic ("tricyclic heterocyclyl") ring systems) and can be saturated or contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups (where the point of attachment is on either the carbocyclyl ring or the heterocyclyl ring), or a heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups (where the point of attachment is on the heterocyclyl ring), in which case the number of ring members is designated through the number of ring members in the heterocyclyl ring system.

[0090] The term "aryl" refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 pi electrons shared among the cyclic array) having 6 to 14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (e.g., having 6, 10, or 14 pi electrons shared among the cyclic array). 6~14 In some embodiments, an aryl group has 6 ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10Aryl"; e.g. naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C 14 "Aryl"; for example, anthracyl. "Aryl" also includes ring systems in which an aryl ring is fused with one or more carbocyclyl or heterocyclyl groups, as defined above, but where the radical or point of attachment is on the aryl ring. In such instances, the number of carbon atoms is designated through the number of carbon atoms in the aryl ring system.

[0091] The term "heteroaryl" refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared among the cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-14 membered heteroaryl"). In some embodiments, the heteroaryl is a substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl, where 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur. In some embodiments, the heteroaryl is a substituted or unsubstituted 9- or 10-membered bicyclic heteroaryl, where 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be at a carbon atom or a nitrogen atom, as valence permits. Heteroaryl polycyclic ring systems may include one or more heteroatoms in one or both rings. "Heteroaryl" encompasses ring systems in which a heteroaryl ring as defined above is fused with one or more carbocyclyl or heterocyclyl groups, and the point of attachment is on the heteroaryl ring, in such cases the number of ring members is designated through the number of ring members in the heteroaryl ring system. "Heteroaryl" also encompasses ring systems in which a heteroaryl ring as defined above is fused with one or more aryl groups, and the point of attachment is on either the aryl ring or the heteroaryl ring, in such cases the number of ring members is designated through the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system. In polycyclic heteroaryl groups in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.), the point of attachment can be on either ring, e.g., on either the ring bearing a heteroatom or the ring that does not contain a heteroatom.

[0092] The addition of the suffix "-ene" to a group indicates that the group is a divalent component, by way of example, alkylene is a divalent component of alkyl, alkenylene is a divalent component of alkenyl, alkynylene is a divalent component of alkynyl, heteroalkylene is a divalent component of heteroalkyl, heteroalkenylene is a divalent component of heteroalkenyl, heteroalkynylene is a divalent component of heteroalkynyl, carbocyclylene is a divalent component of carbocyclyl, heterocyclylene is a divalent component of heterocyclyl, arylene is a divalent component of aryl, and heteroarylene is a divalent component of heteroaryl.

[0093] A chemical moiety is optionally substituted unless expressly provided otherwise. The term "optionally substituted" refers to substituted or unsubstituted. In certain embodiments, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, and acyl groups are optionally substituted. In general, the term "substituted" when referring to a chemical group means that at least one hydrogen present on the group is replaced with an acceptable substituent (e.g., a substituent that upon substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction). Unless otherwise indicated, a "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position is substituted in any given structure, the substituent is either the same or different at each position. The present invention is not limited in any way by the exemplary substituents described herein.

[0094] Exemplary substituents include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa , -ON(R bb )2, -N(R bb )2, -N(R bb )3 + X- 、-N(OR cc )R bb 、-SH、-SR aa 、-SCN、-SSR cc 、-C(=O)R aa 、-CO2H、-CHO、-C(OR cc )2、-CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa, -OC(=O)SR aa , -SC(=O)OR aa , -SC(=O)R aa , -P(=O)(R aa )2, -P(=O)(OR cc )2, -OP(=O)(R aa )2, -OP(=O)(OR cc )2, -P(=O)(N(R bb )2)2, -OP(=O)(N(R bb )2)2, -NR bb P(=O)(R aa )2, -NR bb P(=O)(OR cc )2, -NR bb P(=O)(N(R bb )2)2, -P(R cc )2, -P(OR cc )2, -P(R cc )3 + X - , -P(OR cc )3 + X - , -P(R cc )4, -P(OR cc ) 4, -OP(R cc )2, -OP(R cc )3 + X - , -OP(OR cc )2, -OP(OR cc )3 + X - , -OP(R cc ) 4, -OP(OR cc )4, -B(R aa )2, -B(OR cc )2, -BR aa (OR cc ), C 1~20 Alkyl, C 1~20 Perhaloalkyl, C 1~20 Alkenyl, C 1~20 Alkynyl, Hetero C 1~20 Alkyl, Hetero C 1~20 Alkenyl, Hetero C 1~20 Alkynyl, C 3~10 Carbocyclyl, 3-14 membered heterocyclyl, C 6~14aryl, and 5- to 14-membered heteroaryl; - is the counterion; Alternatively, the two geminal hydrogens on a carbon atom can be replaced by groups =O, =S, =NN(R bb )2, =NNR bb C(=O)R aa , =NNR bb C(=O)OR aa , =NNR bb S(=O)2R aa , =NR bb , or =NOR cc has been replaced by; where: R aa Each of the above may be independently selected from the group consisting of C 1~20 Alkyl, C 1~20 Perhaloalkyl, C 1~20 Alkenyl, C 1~20 Alkynyl, Hetero C 1~20 Alkyl, Hetero C 1~20 Alkenyl, Hetero C 1~20 Alkynyl, C 3~10 Carbocyclyl, 3-14 membered heterocyclyl, C 6~14 aryl, and 5- to 14-membered heteroaryl, or two R aa the groups are joined to form a 3- to 14-membered heterocyclyl ring or a 5- to 14-membered heteroaryl ring; R bb each independently represents hydrogen, -OH, -OR aa , -N(R cc )2, -CN, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NR cc ) OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc, -C(=S)SR cc , -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)(N(R cc )2)2, C 1~20 Alkyl, C 1~20 Perhaloalkyl, C 1~20 Alkenyl, C 1~20 Alkynyl, Hetero C 1~20 Alkyl, Hetero C 1~20 Alkenyl, Hetero C 1~20 Alkynyl, C 3~10 Carbocyclyl, 3-14 membered heterocyclyl, C 6~14 aryl, and 5- to 14-membered heteroaryl, or two R bb the groups are joined to form a 3- to 14-membered heterocyclyl ring or a 5- to 14-membered heteroaryl ring; R cc each independently represents hydrogen, C 1~20 Alkyl, C 1~20 Perhaloalkyl, C 1~20 Alkenyl, C 1~20 Alkynyl, Hetero C 1~20 Alkyl, Hetero C 1~20 Alkenyl, Hetero C 1~20 Alkynyl, C 3~10 Carbocyclyl, 3-14 membered heterocyclyl, C 6~14 aryl, and 5- to 14-membered heteroaryl, or two R cc groups are joined to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring; and each X - is the counter ion.

[0095] In certain embodiments, each substituent is independently a halogen, a substituted (e.g., substituted with one or more halogens) or an unsubstituted C 1~6 Alkyl, -OR aa , -SR aa , -N(R bb )2, -CN, -SCN, -NO2, -N3, -C(=O)R aa , -CO2R aa, -C(=O)N(R bb )2, -OC(=O)R aa , -OCO2R aa , -OC(=O)N(R bb )2, -NR bb C(=O)R aa , -NR bb CO2R aa , or -NR bb C(=O)N(R bb )2.

[0096] The term "halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) or iodine (iodo, -I).

[0097] The term "hydroxyl" or "hydroxy" refers to an -OH group. The term "substituted hydroxyl" or "substituted hydroxy" by extension refers to a hydroxyl group in which the oxygen atom directly attached to the parent molecule has been replaced with a group other than hydrogen, such as -OR aa , -ON(R bb )2, -OC(=O)SR aa , -OC(=O)R aa , -OCO2R aa , -OC(=O)N(R bb )2, -OC(=NR bb )R aa , -OC(=NR bb ) OR aa , -OC(=NR bb )N(R bb )2, -OS(=O)R aa , -OSO2R aa , -OSi(R aa )3, -OP(R cc )2, -OP(R cc )3 + X - , -OP(OR cc )2, -OP(OR cc )3 + X - , -OP(=O)(R aa )2, -OP(=O)(OR cc )2, and -OP(=O)(N(R bb))2, where X - , R aa , R bb , and R cc is as defined herein.

[0098] The term "thiol" or "thio" refers to the group -SH. The term "substituted thiol" or "substituted thio" by extension refers to a group in which the sulfur atom directly attached to the parent molecule is replaced with a group other than hydrogen and is -SR aa , -S-SR cc , -SC(=S)SR aa , -SC(=S)OR aa , -SC(=S) N(R bb )2, -SC(=O)SR aa , -SC(=O)OR aa , -SC(=O)N(R bb )2, and -SC(=O)R aa (where R aa , R bb , and R cc refers to a thiol group, including groups selected from:

[0099] The term "amino" refers to the -NH group. The term "substituted amino" by extension refers to mono-substituted amino, di-substituted amino, or tri-substituted amino. In certain embodiments, a "substituted amino" is a mono-substituted amino group or a di-substituted amino group. The term "mono-substituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is replaced with one hydrogen and one non-hydrogen group, -NH(R bb ), -NHC(=O)R aa , -NHCO2R aa , -NHC(=O)N(R bb )2, -NHC(=NR bb )N(R bb )2, -NHSO2R aa , -NHP(=O)(OR cc )2, and -NHP(=O)(N(R bb 2)2)2 includes groups selected from, where R aa , R bb, and R cc is as defined herein, and wherein -NH(R bb ) group R bb is not hydrogen. The term "disubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is replaced with two groups other than hydrogen, -N(R bb )2, -NR bb C(=O)R aa , -NR bb CO2R aa , -NR bb C(=O)N(R bb )2, -NR bb C(=NR bb )N(R bb )2, -NR bb SO2R aa , -NR bb P(=O)(OR cc )2, and -NR bb P(=O)(N(R bb 2)2)2 includes groups selected from, where R aa , R bb , and R cc is as defined herein, with the proviso that the nitrogen atom directly attached to the parent molecule is not replaced with a hydrogen. The term "trisubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is replaced with three groups, -N(R bb )3 and -N(R bb )3 + X - where R bb and X - is as defined herein.

[0100] The term "acyl" refers to a group having the general formula -C(=O)R aa , -C(=O)OR aa , -C(=O)-OC(=O)R aa , -C(=O)SR aa , -C(=O)N(R bb )2, -C(=S)R aa , -C(=S)N(R bb )2, and -C(=S)S(R aa ), -C(=NRbb )R aa , -C(=NR bb ) OR aa , -C(=NR bb )SR aa , and -C(=NR bb )N(R bb )2(R in the formula aa and R bb refers to a group having a carbon atom (as defined herein). Exemplary acyl groups include aldehydes (-CHO), carboxylic acids (-COH), ketones, acyl halides, esters, amides, imines, carbonates, carbamates, and ureas.

[0101] A "counterion" or "anionic counterion" is a negatively charged group associated with a positively charged group to maintain electronic neutrality. An anionic counterion may be monovalent (e.g., containing one formal negative charge). An anionic counterion may also be multivalent (e.g., containing more than one formal negative charge), such as divalent or trivalent. Exemplary counterions are halide ions (e.g., F - , Cl - , Br - , I - ), NO3 - , ClO4 - , O.H. - , H2PO4 - , HSO4 - , sulfonate ions (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, 10-camphorsulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1-sulfonic acid-5-sulfonic acid, ethane-1-sulfonic acid-2-sulfonic acid, etc.), carboxylate ions (e.g., acetic acid, propanoic acid, benzoic acid, glyceric acid, lactic acid, tartaric acid, glycolic acid, gluconic acid, etc.), BF4 - , PF4 - , PF6 - , AsF6 - , SbF6 - , B[3,5-(CF3)2C6H3]4] - , B(C6F5)4 - , BPh4- , Al(OC(CF3)3)4 - , and carborane anions (e.g., CB 11 H 12 - or (HCB 11 Me5Br6) - Exemplary counterions, which may be multivalent, include CO3 2- , HPO4 2- , PO4 3- , B4O7 2- , SO4 2- , S2O3 2- , carboxylate anions (e.g., tartaric acid, citric acid, fumaric acid, maleic acid, malic acid, malonic acid, gluconic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, salicylic acid, phthalic acid, aspartic acid, glutamic acid, etc.), and carboranes.

[0102] These and other exemplary substituents are described in more detail in the detailed description, examples, and claims. The present invention is not limited in any way by the above exemplary list of substituents. [Brief description of the drawings]

[0103] BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects of the invention and, together with the description, provide non-limiting examples of the invention.

[0104] [Figure 1] Figure 1. Pharmacokinetic profile of subcutaneous injection of StAMP4 (Table A) in mice at 20 mg / kg using various StAMP:lipid ratios (w / w). "Lipid" = DSPE-MPEG (2000).

[0105] [Diagram 2] Figure 2. Efficacy of StAMP26 (Table A) 4 hours after treatment in a P. aeruginosa neutropenic mouse pneumonia model.

[0106] [Diagram 3] Figure 3. Efficacy of StAMP46 (Table A) 4 hours after treatment in a P. aeruginosa neutropenic mouse pneumonia model.

[0107] [Figure 4] Figure 4. Overnight efficacy of StAMP26 and StAMP46 in a P. aeruginosa neutropenic mouse pneumonia model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0108] DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS Provided herein are stapled antimicrobial peptides (StAMPs), pharmaceutical compositions thereof, and kits containing the same. The peptides provided herein have antimicrobial properties and thus can be used to treat and prevent infectious diseases (e.g., bacterial infections) in a subject, kill and / or inhibit the growth of bacteria, etc. Also provided herein are unstapled peptides that can serve as synthetic precursors to the stapled peptides provided herein.

[0109] Stapled Peptides In one aspect, provided herein are stapled (i.e., cross-linked) peptides. In some embodiments, the stapled peptides provided herein are based on the amino acid sequence of the antimicrobial peptide, Magainin II, but include certain modifications that have been found to confer advantageous properties (e.g., improved antimicrobial activity, selectivity for killing gram-negative bacteria, and / or reduced toxicity). ... 1 and X 2 and X 3 and X 4 The endothelial cell includes two staples (i.e., a bridge) connecting the endothelial cell and the endothelial cell.

[0110] Amino acid sequence Provided herein is the amino acid sequence:

number

[0111] In one embodiment, the stapled peptide, or a pharma- ceutically acceptable salt thereof, has the amino acid sequence:

number

[0112] In some embodiments, the stapled peptide provided herein has the following amino acid sequence:

number

[0113] As described herein, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) may include substitutions of one or more amino acids. The amino acids may be independently substituted with any natural or non-natural amino acid, including, but not limited to, those amino acids provided herein. In some embodiments, the amino acid sequence includes substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 amino acids. In some embodiments, the amino acid sequence includes substitutions of one amino acid. In some embodiments, the amino acid sequence includes substitutions of one or two amino acids. In some embodiments, the amino acid sequence includes substitutions of one to three amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of one to four amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of one to five amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of one to six amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of one to seven amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of one to eight amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of 1 to 9 amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of 1 to 10 amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of 1 to 11 amino acids (inclusive).

[0114] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises one to five amino acid substitutions, inclusive, independently at K4, H7, S8, G18, or E19. In some embodiments, the amino acid sequence comprises one to four amino acid substitutions, inclusive, independently at K4, H7, S8, G18, or E19. In some embodiments, the amino acid sequence comprises one to three amino acid substitutions, inclusive, independently at K4, H7, S8, G18, or E19. In some embodiments, the amino acid sequence comprises one or two amino acid substitutions, independently at K4, H7, S8, G18, or E19. In some embodiments, the amino acid sequence comprises one amino acid substitution at H7, S8, G18, or E19.

[0115] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises an amino acid substitution at H7. In some embodiments, the amino acid substitution at H7 is selected from H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, and H7hArg. In some embodiments, the amino acid sequence comprises an H7K substitution. In some embodiments, the amino acid sequence comprises an H7W substitution. In some embodiments, the amino acid sequence comprises an H7Dab substitution. In some embodiments, the amino acid sequence comprises an H7Orn substitution. In some embodiments, the amino acid sequence comprises an H7Dap substitution. In some embodiments, the amino acid sequence comprises an H7R substitution. In some embodiments, the amino acid sequence comprises an H7hArg substitution.

[0116] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises an amino acid substitution at S8. In some embodiments, the amino acid substitution at S8 is selected from S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, and S8hArg. In some embodiments, the amino acid sequence comprises an S8K substitution. In some embodiments, the amino acid sequence comprises an S8W substitution. In some embodiments, the amino acid sequence comprises an S8N substitution. In some embodiments, the amino acid sequence comprises an S8Q substitution. In some embodiments, the amino acid sequence comprises an S8T substitution. In some embodiments, the amino acid sequence comprises an S8Y substitution. In some embodiments, the amino acid sequence comprises an S8Dab substitution. In some embodiments, the amino acid sequence comprises an S8Orn substitution. In some embodiments, the amino acid sequence comprises an S8Dap substitution. In some embodiments, the amino acid sequence comprises an S8R substitution. In some embodiments, the amino acid sequence comprises an S8hArg substitution.

[0117] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises an amino acid substitution at both H7 and S8. In some embodiments, the amino acid sequence comprises an H7K substitution and an S8K substitution.

[0118] In some embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) include an amino acid substitution at G18. In some embodiments, the amino acid substitution at G18 is G18W, G18V, G18L, G18Y, G18F, G18T, G18F 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 In some embodiments, the amino acid sequence comprises a G18W substitution. In some embodiments, the amino acid sequence comprises a G18V substitution. In some embodiments, the amino acid sequence comprises a G18L substitution. In some embodiments, the amino acid sequence comprises a G18Y substitution. In some embodiments, the amino acid sequence comprises a G18F substitution. In some embodiments, the amino acid sequence comprises a G18T substitution. In some embodiments, the amino acid sequence comprises a G18F 1 In one embodiment, the amino acid sequence comprises the substitution G18F 2 In one embodiment, the amino acid sequence comprises the substitution G18F 4 In one embodiment, the amino acid sequence comprises the substitution G18F 5 In one embodiment, the amino acid sequence comprises the substitution G18F 3 In some embodiments, the amino acid sequence comprises a G18S substitution.

[0119] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises an amino acid substitution at E19. In some embodiments, the amino acid substitution at E19 is selected from E19W, E19D, E19Q, and E19N. In some embodiments, the amino acid sequence comprises an E19W substitution. In some embodiments, the amino acid sequence comprises an E19D substitution. In some embodiments, the amino acid sequence comprises an E19Q substitution. In some embodiments, the amino acid sequence comprises an E19N substitution.

[0120] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) comprises an amino acid substitution at K4. In some embodiments, the amino acid substitution at K4 is selected from K4S, K4Dab, K4Orn, K4Dap, K4R, and K4hArg. In some embodiments, the amino acid sequence comprises a K4S substitution. In some embodiments, the amino acid sequence comprises a K4Dab substitution. In some embodiments, the amino acid sequence comprises a K4Orn substitution. In some embodiments, the amino acid sequence comprises a K4Dap substitution. In some embodiments, the amino acid sequence comprises a K4R substitution. In some embodiments, the amino acid sequence comprises a K4hArg substitution.

[0121] In some embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) comprise an amino acid substitution at G13. In some embodiments, the amino acid substitution at G13 is G13S.

[0122] In some embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) comprise an amino acid substitution at A15. In some embodiments, the amino acid substitution at A15 is A15S.

[0123] In some embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) comprise an amino acid substitution at A21. In some embodiments, the amino acid substitution at A21 is A21S.

[0124] The amino acid sequences provided herein (eg, SEQ ID NO: 1 or 2) can contain any combination of the above amino acid substitutions.

[0125] In certain embodiments, the stapled peptides provided herein, or pharma- ceutically acceptable salts thereof, comprise one of the following amino acid sequences:

number

number

number

[0126] In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus of the peptide is amidated with -NH2. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof, wherein X is an amino acid sequence selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 29, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 82, 83, 84, 85, 86, 87, 88, 89, 1 and X 2 and X 3 and X 4 and X are connected to form a bridge represented by the formula (alk). In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus of the peptide is amidated with -NH; and X 1 and X 2 and X 3 and X 4 are connected to form a bridge represented by the formula (alk).

[0127] In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus of the peptide is amidated with -NH2. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof, wherein X is an amino acid sequence selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 1 and X 2and X 3 and X 4 and X are connected to form a bridge represented by the formula (alk). In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salts thereof, wherein the C-terminus of the peptide is amidated with -NH2; and X 1 and X 2 and X 3 and X 4 are connected to form a bridge represented by the formula (alk).

[0128] Also provided herein is the amino acid sequence:

number

[0129] In an embodiment, the amino acid sequence represented by SEQ ID NO:2 includes substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 amino acids. In an embodiment, the amino acid sequence includes substitutions of 1 amino acid. In an embodiment, the amino acid sequence includes substitutions of 1 or 2 amino acids. In an embodiment, the amino acid sequence includes substitutions of 1 to 3 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 4 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 5 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 6 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 7 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 8 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 9 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 10 amino acids (including both ends). In an embodiment, the amino acid sequence includes substitutions of 1 to 11 amino acids (including both ends). The amino acid substitutions may be selected from any of the substitutions provided herein.

[0130] In some embodiments, the amino acid sequence represented by SEQ ID NO:2 contains one amino acid replaced by W. In some embodiments, the amino acid sequence contains two amino acids replaced by W.

[0131] In some embodiments, the stapled peptide provided herein has the following amino acid sequence:

number

[0132] In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH2. In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof, wherein X is a nucleotide. 1 and X 2 and X 3 and X 4 and X are connected to form a bridge represented by the formula (alk). In some embodiments, the stapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salts thereof, wherein the C-terminus is amidated with -NH2; and X 1 and X 2 and X 3 and X 4 are connected to form a bridge represented by the formula (alk).

[0133] Also provided herein is the amino acid sequence:

number

[0134] In some embodiments, the amino acid sequence represented by SEQ ID NO:3 includes substitutions of 1, 2, 3, 4, or 5 amino acids. In some embodiments, the amino acid sequence includes substitutions of 1 amino acid. In some embodiments, the amino acid sequence includes substitutions of 1 or 2 amino acids. In some embodiments, the amino acid sequence includes substitutions of 1 to 3 amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of 1 to 4 amino acids (inclusive). In some embodiments, the amino acid sequence includes substitutions of 1 to 5 amino acids (inclusive). The amino acid substitutions may be selected from any substitution provided herein.

[0135] In some embodiments, the stapled peptide provided herein comprises SEQ ID NO: 3 or a pharma- ceutically acceptable salt thereof. In some embodiments, the stapled peptide provided herein is represented by SEQ ID NO: 3 or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH2. In some embodiments, the stapled peptide provided herein is represented by SEQ ID NO: 3 or a pharma- ceutically acceptable salt thereof, wherein X is amidated with -NH2. 1 and X 2 and X 3 and X 4 and X are connected to form a bridge represented by the formula (alk). In some embodiments, the stapled peptide provided herein is represented by SEQ ID NO: 3, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH; and X 1 and X 2 and X 3 and X 4 are connected to form a bridge represented by the formula (alk).

[0136] Also provided herein is the amino acid sequence:

number

[0137] In an embodiment, the amino acid sequence represented by SEQ ID NO: 101 includes substitutions of 1, 2, 3, 4, or 5 amino acids. In an embodiment, the amino acid sequence includes substitutions of 1 amino acid. In an embodiment, the amino acid sequence includes substitutions of 1 or 2 amino acids. In an embodiment, the amino acid sequence includes substitutions of 1 to 3 amino acids (inclusive). In an embodiment, the amino acid sequence includes substitutions of 1 to 4 amino acids (inclusive). In an embodiment, the amino acid sequence includes substitutions of 1 to 5 amino acids (inclusive). The amino acid substitutions may be selected from any substitution provided herein.

[0138] In some embodiments, the stapled peptide provided herein has the following amino acid sequence:

number

[0139] In some embodiments, the stapled peptide provided herein comprises any one of SEQ ID NOs: 101-105, or a pharma- ceutically acceptable salt thereof. In some embodiments, the stapled peptide provided herein is represented by ..., wherein the C-terminus is amidated with -NH2. In some embodiments, the stapled peptide provided herein is represented by any one of SEQ ID NOs: 101-105, or a pharma- ceutically acceptable salt thereof, wherein X is an amino acid sequence selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 1 and X 2 and X are connected to form a bridge represented by the formula (alk). In some embodiments, the stapled peptide provided herein is represented by any one of SEQ ID NOs: 101-105, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH2; and X 1 and X 2 are connected to form a bridge represented by the formula (alk).

[0140] In addition to pharma- ceutically acceptable salts of the stapled peptides provided herein, also provided herein are stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs of the stapled peptides provided herein.

[0141] As described herein, stapling (e.g., cross-linking) a peptide can stabilize the secondary structure (e.g., the secondary structure of an α-helix) of the peptide. In some embodiments, one or more cross-links of a stapled peptide provided herein stabilize the α-helix of the peptide. In some embodiments, the peptide has increased α-helicity compared to the corresponding unstapled (e.g., non-cross-linked) peptide.

[0142] The stapled peptides provided herein may exhibit α-helical stability by maintaining the structure of an α-helical structure as measured by circular dichroism or NMR. For example, in some embodiments, the stapled peptide exhibits at least a 1.1, 1.2, 1.25, 1.3, 1.4, 1.5, 1.6, 1.7, 1.75, 1.8, 1.9, or 2-fold increase in α-helicity (e.g., as measured by circular dichroism or NMR) compared to the corresponding unstapled peptide. In certain embodiments, the stapled peptides provided herein may exhibit about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% α-helicity compared to the corresponding unstapled peptide (e.g., as measured by circular dichroism or NMR).

[0143] Additional amino acid substitutions Another aspect of the present disclosure relates to the discovery that replacing lysine (K) and / or phenylalanine (F) residues of the amino acid sequences listed herein with modified variants can serve to impart advantageous properties (e.g., improved antimicrobial activity, reduced toxicity, etc.). For example, it has been found that replacing one or more lysine (K) residues with "shorter lysine" residues (e.g., amino acids Orn, Dab, and Dap shown in Table 2 below) can serve to reduce the toxicity (e.g., nephrotoxicity) of the peptide. Table 2. "Shorter Lysine" Amino Acids [Table 2]

[0144] Thus, also provided herein are peptides and pharma- ceutically acceptable salts thereof comprising any of the amino acid sequences provided herein (e.g., SEQ ID NOs: 1-53, 59-77, and 81-105), optionally wherein one or more of K are independently substituted with an amino acid selected from Orn, Dab, Dap, R, and hArg. In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are independently substituted with an amino acid selected from Orn, Dab, and Dap. In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are substituted with Orn. ​​In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are substituted with Dab. In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are replaced by Dap. In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are replaced by R. In some embodiments, one or more of K (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 K) are replaced by hArg.

[0145] In some embodiments, each K is independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg. In some embodiments, each K is replaced by Orn. ​​In some embodiments, each K is replaced by Dab. In some embodiments, each K is replaced by Dap. In some embodiments, each K is replaced by R. In some embodiments, each K is replaced by hArg.

[0146] Also provided herein are peptides, and pharma- ceutically acceptable salts thereof, comprising any of the amino acid sequences provided herein (e.g., SEQ ID NOs: 1-53, 59-77, and 81-105), optionally wherein one or more of the F's (e.g., each of the F's) are independently replaced by a modified phenylalanine. "Modified phenylalanine," for purposes of this disclosure, refers to a stereoisomer of phenylalanine (e.g., D-Phe) or any non-natural phenylalanine analog, in which the phenyl ring is ortho-, meta-, and / or para-substituted with one or more non-hydrogen substituents (e.g., optionally substituted alkyl (e.g., Me, Et, n-Pr, i-Pr, n-Bu, t-Bu), halogen (e.g., F, Cl, Br, I), haloalkyl (e.g., -CF), optionally substituted hydroxyl (e.g., -OH), optionally substituted amino (e.g., -NH), optionally substituted thio (e.g., -SH), optionally substituted acyl (e.g., -C(=O)Me, -C(=O)OH, -C(=O)NH), -CN, -SCN, -NO, -N). In some embodiments, one or more of the F's are independently replaced by a modified phenylalanine. In certain embodiments, each F is independently replaced by a modified phenylalanine.

[0147] In some embodiments, one or more of F (e.g., one, two, three of F) are independently selected from the group consisting of F, ... 1 , F 5 , F 4 , F 2 , and F 3 In some embodiments, one or more of the F's (e.g., one, two, three of the F's) are independently substituted with an amino acid selected from the group consisting of F 1 In some embodiments, one or more of F (e.g., one, two, three of F) are independently substituted by F. 5 In some embodiments, one or more of F (e.g., one, two, three of F) are independently substituted by F. 4 In some embodiments, one or more of F (e.g., one, two, three of F) are independently substituted by F.2 In some embodiments, one or more of F (e.g., one, two, three of F) are independently substituted by F. 3 has been replaced by

[0148] In some embodiments, each F is 1 , F 5 , F 4 , F 2 , and F 3 In one embodiment, each F is substituted with an amino acid selected from 1 In some embodiments, each F is substituted by F 5 In some embodiments, each F is substituted by F 4 In some embodiments, each F is substituted by F 2 In some embodiments, each F is substituted by F 3 has been replaced by

[0149] Also provided herein are peptides, and pharma- ceutically acceptable salts thereof, comprising any of the amino acid sequences provided herein (e.g., SEQ ID NOs: 1-53, 59-77, and 81-105), optionally wherein one or more of K are independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg; and optionally wherein one or more of F are independently replaced by a modified phenylalanine.

[0150] All combinations of the above amino acid substitutions (ie, H7 substitutions, S8 substitutions, G18 substitutions, E19 substitutions, K substitutions, F substitutions, and others) are encompassed by the present disclosure.

[0151] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) may include, independently, one to five (e.g., one, two, four, three, four, five) amino acid substitutions (inclusive), such as K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F), G18F (e.g., G18F ... 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), or E19 (e.g., E19W, E19D, E19Q, or E19N); optionally, further wherein one or more of K (e.g., each of K) are independently substituted with an amino acid selected from Orn, Dab, Dap, R, and hArg; and optionally, further wherein one or more of F (e.g., each of F) are independently substituted with a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , or F 3 ) has been substituted.

[0152] In some embodiments, the amino acid sequence provided herein (e.g., SEQ ID NO: 1 or 2) may include, independently, one to five (e.g., one, two, four, three, four, five) amino acid substitutions (inclusive), such as K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G13 (e.g., G13S), A15 (e.g., A15S), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F), G18F (e.g., G18F ... 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), E19 (e.g., E19W, E19D, E19Q, or E19N), or A21 (e.g., A21S); optionally, further wherein one or more of K (e.g., each of K) are independently substituted with an amino acid selected from Orn, Dab, Dap, R, and hArg; and optionally, further wherein one or more of F (e.g., each of F) are independently substituted with a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , or F 3 ) has been substituted.

[0153] In some embodiments, the amino acid sequences provided herein (e.g., SEQ ID NOs: 1, 2, or 3) include one or two amino acids substituted by W; optionally, further wherein one or more of K (e.g., each of K) are independently substituted by an amino acid selected from Orn, Dab, Dap, R, and hArg; and optionally, further wherein one or more of F (e.g., each of F) are independently substituted by a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , and F3 ) has been substituted.

[0154] In certain embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) can include any combination of the following amino acid substitutions: (i) independently, K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), or a substitution of 1 to 5 (e.g., 1, 2, 4, 3, 4, 5) amino acids (inclusive) at E19 (e.g., E19W, E19D, E19Q, or E19N); (ii) one or more of the Ks (e.g., each K) are independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg; and / or (iii) one or more of F (e.g., each of F) is independently a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , and F 3 ) has been substituted.

[0155] In certain embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) can include any combination of the following amino acid substitutions: (i) independently, K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), or a substitution of 1 to 5 (e.g., 1, 2, 4, 3, 4, 5) amino acids (inclusive) at E19 (e.g., E19W, E19D, E19Q, or E19N); (ii) 1 or 2 amino acids replaced by W; (iii) one or more of the Ks (e.g., each K) are independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg; and / or (iv) one or more of F (e.g., each of F) is independently a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , and F 3 ) has been substituted.

[0156] In certain embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) can include any combination of the following amino acid substitutions: (i) independently, K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G13 (e.g., G13S), A15 (e.g., A15S), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), E19 (e.g., E19W, E19D, E19Q, or E19N), or A21 (e.g., A21S) substitution of 1 to 5 (e.g., 1, 2, 4, 3, 4, 5) amino acids (inclusive); (ii) one or more of the Ks (e.g., each K) are independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg; and / or (iii) one or more of F (e.g., each of F) is independently a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , and F 3 ) has been substituted.

[0157] In certain embodiments, the amino acid sequences provided herein (e.g., SEQ ID NO: 1 or 2) can include any combination of the following amino acid substitutions: (i) independently, K4 (e.g., K4S, K4Dab, K4Orn, K4Dap, K4R, or K4hArg), H7 (e.g., H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, or H7hArg), S8 (e.g., S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, or S8hArg), G13 (e.g., G13S), A15 (e.g., A15S), G18 (e.g., G18W, G18V, G18L, G18Y, G18F, G18T, G18F 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 , or G18S), E19 (e.g., E19W, E19D, E19Q, or E19N), or A21 (e.g., A21S) substitution of 1 to 5 (e.g., 1, 2, 4, 3, 4, 5) amino acids (inclusive); (ii) 1 or 2 amino acids replaced by W; (iii) one or more of the Ks (e.g., each K) are independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg; and / or (iv) one or more of F (e.g., each of F) is independently a modified phenylalanine (e.g., F 1 , F 5 , F 4 , F 2 , and F 3 ) has been substituted.

[0158] Stapled peptide crosslinking The stapled peptides provided herein include bridges (e.g., staples), where each bridge connects two amino acids (i.e., the amino acids being bridged) to form a macrocycle. In some embodiments, the amino acid sequence is X 1 and X 2 When contains X 1 and X 2 are bridged amino acids connected via a bridge. Also, in some embodiments, the amino acid sequence X3 and X 4 When contains X 3 and X 4 are bridged amino acids that are connected via a bridge.

[0159] In some embodiments, the bridges are independently located at the alpha positions of the bridged amino acids (e.g., X 1 , X 2 , X 3 , and X 4 In some embodiments, the bridges are attached to the alpha position of the bridged amino acid (e.g., X 1 , X 2 , X 3 , and X 4 ) and the bridged amino acids are independently α,α-disubstituted amino acids.

[0160] In certain embodiments, each bridge is independently from about 5 Å to about 25 Å in length, inclusive. In certain embodiments, each bridge is independently from about 6 Å to about 22 Å in length, inclusive. In certain embodiments, each bridge is independently from about 7 Å to about 20 Å in length, inclusive. In certain embodiments, each bridge is independently from about 8 Å to about 18 Å in length, inclusive. In certain embodiments, each bridge is independently from about 9 Å to about 17 Å in length, inclusive. In certain embodiments, each bridge is independently from about 10 Å to about 16 Å in length, inclusive. In certain embodiments, each bridge is independently from about 11 Å to about 15 Å in length, inclusive. In certain embodiments, each bridge is independently from about 12 Å to about 14 Å in length, inclusive. In certain embodiments, each bridge is independently about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 Å in length.

[0161] In some embodiments, the length of each bridge is approximately equal to the length of 5-25 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 5-20 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 5-15 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 5-13 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 6-12 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 7-11 carbon-carbon bonds (inclusive). In some embodiments, the length of each bridge is approximately equal to the length of 8-10 carbon-carbon bonds (inclusive). In certain embodiments, the length of each bridge is approximately equal to the length of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 carbon-carbon bonds (inclusive).

[0162] In some embodiments, at least one bridge spans at least one turn of the α-helix of the peptide. In some embodiments, each bridge spans at least one turn of the α-helix of the peptide. In some embodiments, at least one bridge spans at least one turn of the α-helix of the peptide. In some embodiments, each bridge spans at least one turn of the α-helix of the peptide.

[0163] In some embodiments, each pair of cross-linked amino acids (e.g., X 1 and X 2 and X 3 and X 4 and ) are independently connected by a bridge to form a compound having the following formula: [ka] where α represents the α-carbon of the bridged amino acid; 1 is a bridge; and R 1 Each of is independently hydrogen or an optionally substituted C1~6 It forms an alkyl group.

[0164] In some embodiments, each bridge (e.g., L 1 ) are independently optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, optionally substituted acylene, or any combination thereof.

[0165] In some embodiments, each bridge (e.g., L 1 ) is independently a hydrocarbon bridge. For purposes of this disclosure, a "hydrocarbon bridge" is a bridge consisting of an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, and combinations thereof.

[0166] In some embodiments, each bridge (e.g., L 1 ) is independently an optionally substituted alkenylene (e.g., unsubstituted alkenylene). In certain embodiments, each bridge independently has the formula: [ka] where each n is independently an integer from 1 to 10, inclusive. In certain embodiments, the sum of the two n's on the same bridge is 6.

[0167] In some embodiments, the cross-linked amino acids (e.g., X 1 , X 2 , X 3 , and X 4 ) are independently α,α-disubstituted amino acids. Illustratively, in some embodiments, each pair of bridged amino acids (e.g., X 1 and X 2 and X 3 and X4 and ) are independently connected by bridges to form the following formula: [ka] where α represents the α-carbon of the bridged amino acid; and where R 1 each independently represents an optionally substituted C 1~6 In certain embodiments, the sum of the two n's on the same bridge is 6.

[0168] For example, in some embodiments, crosslinking (e.g., L 1 ) are independently selected from the formula: [ka] It is expressed as:

[0169] For example, in one embodiment, a pair of cross-linked amino acids (e.g., X 1 and X 2 and X 3 and X 4 and ) are independently connected via a bridge to form a compound having the following formula: [ka] where α represents the α-carbon of the bridged amino acid.

[0170] In one embodiment, X 1 and X 2 are connected to form an expression (alk).

[0171] In one embodiment, X 3 and X 4 are connected to form an expression (alk).

[0172] In some embodiments, crosslinking (e.g., L 1) is independently an optionally substituted alkylene (e.g., unsubstituted alkylene). In certain embodiments, each bridge independently has the formula: [ka] wherein m is an integer from 1 to 20, inclusive. In one embodiment, m is 6.

[0173] In some embodiments, a pair of cross-linked amino acids (e.g., X 1 and X 2 and X 3 and X 4 and) are independently linked by bridges to form a compound of the formula: [ka] where α represents the α-carbon of the bridged amino acid; and where R 1 each independently represents an optionally substituted C 1~6 In some embodiments, m is 6.

[0174] For example, in some embodiments, crosslinking (e.g., L 1 ) are independently selected from the formula: [ka] It is expressed as:

[0175] For example, in one embodiment, a pair of cross-linked amino acids (e.g., X 1 and X 2 and X 3 and X 4 and ) are connected via a bridge to form the following formula: [ka] where α represents the α-carbon of the bridged amino acid.

[0176] In some embodiments, crosslinking (e.g., L 1 ) is independently an optionally substituted alkynylene (eg, unsubstituted alkynylene).

[0177] In some embodiments, crosslinking (e.g., L 1 ) are independently a dithio bridge. For purposes of this disclosure, a "dithio bridge" is a bridge that contains two thioethers (i.e., two -S- groups). In certain embodiments, the bridges are independently a dithio bridge represented by the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive; L 2 is optionally substituted alkylene, [ka] , optionally substituted arylene, optionally substituted heteroarylene, or -A 1 -A 1 -; 1 Each of is independently an optionally substituted arylene or an optionally substituted heteroarylene. In certain embodiments, each n is 1.

[0178] In certain embodiments, the bridges independently have the formula: [ka] In some embodiments, each n is 1.

[0179] For example, in certain embodiments, the bridges independently have the formula: [ka] It is a dithio bridge represented by one of the following formulas:

[0180] In other embodiments, the bridges are independently dithio bridges represented by the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive; L 2 is an optionally substituted aromatic ring (e.g., a polyhalogenated aryl or heteroaryl ring) or -A 1 -A 1 - is; where A 1 Each of is independently an optionally substituted aromatic ring (e.g., a polyhalogenated aryl or heteroaryl ring). In some embodiments, each n is 1.

[0181] In certain embodiments, the bridges independently have the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive. In some embodiments, each n is 1.

[0182] For example, in certain embodiments, the bridges independently have the formula: [ka] It is expressed as:

[0183] In some embodiments, crosslinking (e.g., L 1 ) is independently a triazolylene bridge. For purposes of this disclosure, a "triazolylene bridge" refers to at least one triazolylene moiety (e.g., [ka] It is a bridge interrupted by.

[0184] In certain embodiments, the bridges independently have the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive. In certain embodiments, the sum of the two n's on the same bridge is 5.

[0185] For example, in certain embodiments, the bridges independently have the formula: [ka] It is a triazolylene bridge represented by one of the following formulas:

[0186] n and R 1 The following aspects apply to all general formulas and subgenera provided herein, and to all stapled and unstapled peptides provided herein.

[0187] In certain embodiments, the sum of two n's on the same bridge is an integer from 3 to 9, inclusive. In some embodiments, the sum of the two n's on the same bridge is an integer from 4 to 8, inclusive. In some embodiments, the sum of the two n's on the same bridge is an integer from 5 to 7, inclusive. In some embodiments, the sum of the two n's on the same bridge is 5. In some embodiments, the sum of the two n's on the same bridge is 6. In some embodiments, the sum of the two n's on the same bridge is 7.

[0188] In some embodiments, at least one of n is 1. In some embodiments, at least one of n is 2. In some embodiments, at least one of n is 3. In some embodiments, at least one of n is 4. In some embodiments, at least one of n is 5. In some embodiments, at least one of n is 6. In some embodiments, at least one of n is 7. In some embodiments, at least one of n is 8. In some embodiments, at least one of n is 9. In some embodiments, at least one of n is 10.

[0189] In some embodiments, m is an integer from 3 to 9 (inclusive). In some embodiments, m is an integer from 4 to 8 (inclusive). In some embodiments, m is an integer from 5 to 7 (inclusive). In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 7.

[0190] In one embodiment, R 1 At least one of R is hydrogen. 1 Each of R is hydrogen. 1 At least one of the groups is unsubstituted C 1~6 In some embodiments, R 1 At least one of the groups is unsubstituted C 1~3 In some embodiments, R 1 At least one of R is methyl. 1 Each of is methyl.

[0191] Herein, the recitation of a list of chemical groups in any definition of a variable includes that definition of the variable as any single group or combination of groups in the list. Herein, the recitation of an embodiment of a variable includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. Herein, the recitation of an embodiment includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0192] Peptide C-terminal modification The peptides provided herein may include one or more additional modifications anywhere on the peptide (e.g., on the side chain of an amino acid, on the α-carbon of an amino acid, on the peptidic nitrogen, at the C-terminus, at the N-terminus, etc.). The peptides provided herein may include modifications to the C-terminus and / or N-terminus of the polypeptide. In some embodiments, the polypeptide includes a modified C-terminus. Examples of C-terminal modifications are described herein.

[0193] In some embodiments, the peptides provided herein include an amidated C-terminus. Traditionally, peptides include a carboxyl group (-C(=O)OH) at the C-terminus. The peptides provided herein may include an amide at the C-terminus (e.g., -C(=O)NR2, where the group NR2 is NH2, mono-substituted amino, di-substituted amino, or tri-substituted amino) and are referred to as an "amidated C-terminus." For example, a peptide with an "-NH2-amidated C-terminus" includes the group -C(=O)NH2 at the C-terminus in place of a carboxyl (-C(=O)OH). An amidated C-terminus can also be represented by the inclusion of -NR2 (e.g., -NH2) at the C-terminus of the amino acid sequence.

[0194] The peptides provided herein may also be amidated at the C-terminus with an amino acid, peptide, or protein. The amino acid, peptide, or protein may be natural or non-natural. In some embodiments, the peptide comprises a peptide conjugated to the C-terminus. In some embodiments, the peptide is 2 to 6 amino acids in length (inclusive) and comprises amino acids selected from G, E, and S. In some embodiments, the peptide is 2 to 6 amino acids in length (inclusive) and comprises amino acids selected from G and E. In some embodiments, the peptide is 2 amino acids in length and comprises amino acids selected from G and E. In some embodiments, the peptide is 3 amino acids in length and comprises amino acids selected from G and E. In some embodiments, the peptide is 4 amino acids in length and comprises amino acids selected from G and E.

[0195] Non-limiting examples of peptides that can be conjugated to the C-terminus are:

number

[0196] The peptides provided herein may also include a small molecule, lipophilic group, or polymer conjugated to the C-terminus of the peptide. In some embodiments, the peptide includes a lipophilic group conjugated to the C-terminus of the peptide. In some embodiments, the lipophilic group is a lipid or a fatty acid. In some embodiments, the lipophilic group is a hydrocarbon chain.

[0197] In some embodiments, the peptide comprises a polymer conjugated to the C-terminus of the peptide. In some embodiments, the polymer is a polyether, such as polyethylene glycol (PEG). In some embodiments, the polymer is PEG. In some embodiments, the C-terminus comprises PEG3. In some embodiments, the C-terminus comprises (PEG3)2. In some embodiments, the C-terminus comprises -NH(PEG). In some embodiments, the C-terminus comprises -NH(PEG3). In some embodiments, the C-terminus comprises -NH(PEG3)2.

[0198] In some embodiments, the peptide is amidated at the C-terminus with a group represented by the formula: -NH-(PEG)-CONH2, where PEG is polyethylene glycol. In some embodiments, the peptide is amidated at the C-terminus with a group represented by the formula: -NH(CH2CH2O) 1~20 In some embodiments, the polymer is amidated at the C-terminus with a group represented by one of the following formulas: -NHCH2CH2OCH2CH2CONH2, -NH(CH2CH2O)2-CH2CH2CONH2, -NH(CH2CH2O)3-CH2CH2CONH2, -NH(CH2CH2O)4-CH2CH2CONH2, or -NH(CH2CH2O)5-CH2CH2CONH2.

[0199] In some embodiments, the peptide comprises a small molecule conjugated to the C-terminus of the peptide. In some embodiments, the small molecule is an antimicrobial agent, such as a small molecule antibiotic. In some embodiments, the small molecule is a cyclic antimicrobial peptide conjugated to the C-terminus. Polymyxin is a non-limiting example of a cyclic antimicrobial peptide.

[0200] In some embodiments, the stapled peptide, or a pharma- ceutically acceptable salt thereof, has the following amino acid sequence with the corresponding C-terminal modification:

number

[0201] Unstapled peptides Also provided herein are unstapled (i.e., non-crosslinked) peptides that can serve as synthetic precursors to the stapled (i.e., crosslinked) peptides provided herein. Such unstapled peptides may also be referred to as "unstapled precursor peptides," "unstapled precursors," and the like. Unstapled precursor peptides include pairs of amino acids that include reactive moieties that can react to form crosslinks.

[0202] Amino acid sequences and substitutions Provided herein is the amino acid sequence:

number

[0203] In some embodiments, the unstapled peptide or a pharma- ceutically acceptable salt thereof has the amino acid sequence:

number

[0204] In some embodiments, the unstapled peptides provided herein have the following amino acid sequence:

number

[0205] Because the unstapled peptides provided herein can serve as synthetic precursors to the stapled peptides provided herein, the various numbers, positions, and types of amino acid substitutions described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein. In addition, the possible C-terminal modifications described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein.

[0206] In some embodiments, the unstapled peptides provided herein comprise one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 4-39 or 59-77, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus of the peptide is amidated with -NH2.

[0207] In some embodiments, the unstapled peptides provided herein comprise one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 81-100, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus of the peptide is amidated with -NH2.

[0208] Also provided herein is the amino acid sequence:

number

[0209] The various numbers, positions, and types of amino acid substitutions described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein. The possible C-terminal modifications described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein.

[0210] In some embodiments, the unstapled peptides provided herein comprise one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by one of SEQ ID NOs: 40-53, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH2.

[0211] Also provided herein is the amino acid sequence:

number

[0212] As described herein, the various numbers, positions, and types of amino acid substitutions described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein. The possible C-terminal modifications described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein.

[0213] In some embodiments, the unstapled peptides provided herein comprise SEQ ID NO: 3. In some embodiments, the unstapled peptides provided herein are represented by SEQ ID NO: 3, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by SEQ ID NO: 3, or a pharma- ceutically acceptable salt thereof, where the C-terminus is amidated with -NH2.

[0214] Also provided herein is the amino acid sequence:

number

[0215] As described herein, the various numbers, positions, and types of amino acid substitutions described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein. The possible C-terminal modifications described herein for the stapled peptides (e.g., above) also apply to the unstapled peptides provided herein.

[0216] In some embodiments, the unstapled peptides provided herein comprise any one of SEQ ID NOs: 101-105. In some embodiments, the unstapled peptides provided herein are represented by any one of SEQ ID NOs: 101-105, or a pharma- ceutically acceptable salt thereof. In some embodiments, the unstapled peptides provided herein are represented by any one of SEQ ID NOs: 101-105, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with -NH2.

[0217] In addition to pharma- ceutically acceptable salts of the stapled peptides provided herein, also provided herein are stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs of the stapled peptides provided herein.

[0218] Reactive moieties As described herein, an unstapled (i.e., uncrosslinked) peptide comprises a pair of amino acids (e.g., X) that contain reactive moieties capable of forming crosslinks. 1 and X 2 and X 3 and X 4 In a preferred embodiment, the amino acid pair (e.g., X 1 and X 2 and X 3and X 4 with) comprises an α-side chain that contains a reactive moiety.

[0219] For purposes of this disclosure, a "reactive moiety" is any chemical moiety that can react with another chemical moiety to form a covalent bond or bonds. Non-limiting examples of reactive moieties include alkenes, alkynes, alcohols, amines, thiols, azides, esters, amides, halogens, and the like. In some embodiments, two reactive moieties can react directly with each other to form a crosslink (e.g., an alkene via ring-closing metathesis (RCM), or an alkyne and an azide via 1,3-dipolar cycloaddition to form a triazole). In other embodiments, two reactive moieties react with an intermediate crosslinking reagent to form a crosslink (e.g., two cysteine ​​residues react with a dihalide, e.g., dibromoxylene). In some embodiments, the reactive moiety is a click chemistry moiety. A "click chemistry" moiety is any moiety that can be used in a click chemistry reaction (described below).

[0220] In some embodiments, the reactive moieties are independently selected from alkenes and alkynes (e.g., those capable of undergoing an RCM reaction to form a hydrocarbon bridge). In some embodiments, the reactive moiety is a terminal alkene. In some embodiments, the reactive moiety is a terminal alkyne.

[0221] In one embodiment, X 1 , X 2 , X 3 , and X 4 are independently of the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive. In some embodiments, at least one of the n's is 3. In some embodiments, the sum of the two n's between the bridging amino acids is 6.

[0222] In one embodiment, X 1 , X 2 , X 3 , and X 4 is independently an α,α-disubstituted amino acid. 1 , X 2 , X 3 , and X 4 are each independently represented by the formula: [ka] where α represents the α-carbon of an amino acid; and R 1 Each of the groups may be an optionally substituted C 1~6 In some embodiments, R 1 is methyl.

[0223] In one embodiment, at least one of X 1 , X 2 , X 3 , and X 4 is the following formula: [ka] In the formula, each n is independently an integer of 1 to 10 (inclusive), and is an amino acid represented by the formula:

[0224] In one embodiment, at least one of X 1 , X 2 , X 3 , and X 4 is amino acid S 5 : [ka] It is.

[0225] In one embodiment, X 1 is S 5 In one embodiment, X 2 is S 5 In one embodiment, X 3is S 5 In one embodiment, X 4 is S 5 In one embodiment, X 1 , X 2 , X 3 , and X 4 is S 5 It is.

[0226] In one embodiment, X 1 is R 8 In one embodiment, X 2 is R 8 In one embodiment, X 3 is R 8 In one embodiment, X 4 is R 8 In one embodiment, X 1 , X 2 , X 3 , and X 4 is R 8 It is.

[0227] In some embodiments, the reactive moiety is a thiol (e.g., capable of reacting with a cross-linking reagent, such as a dihalide (e.g., dibromoxylene), to form a dithio cross-link). 1 , X 2 , X 3 , and X 4 are independently of the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive. In some embodiments, at least one n is 1. In some embodiments, each n is 1.

[0228] In one embodiment, X 1 , X 2 , X 3 , and X 4 Each of X is cysteine ​​(C). 1 and X 2is C. In some embodiments, X 3 and X 4 is C.

[0229] In some embodiments, the reactive moieties are azides and alkynes (e.g., those that can react via 1,3-cycloaddition to form triazolylene bridges). 1 and X 2 One of X contains an alkyne reactive moiety and the other contains an azide reactive moiety. 3 and X 4 One of them contains an alkyne reactive moiety and the other contains an azide reactive moiety.

[0230] In one embodiment, X 1 and X 2 One of the formulas is: [ka] and the other has the formula: [ka] In one embodiment, X 3 and X 4 One of the formulas is: [ka] and the other has the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive. In one embodiment, X 1 and X 2 One of the formulas is: [ka] and the other has the formula: [ka] In one embodiment, X 3 and X 4 One of the formulas is: [ka] and the other has the formula: [ka] The α-side chain is represented by

[0231] In one embodiment, X 1 and X 2 One of X is J and the other is Azi. 3 and X 4 One of them is J and the other is Azi.

[0232] Methods for preparing stapled peptides Also provided herein are methods of preparing the cross-linked (i.e., stapled) peptides described herein, the methods comprising reacting non-cross-linked (i.e., unstapled) peptides under conditions sufficient to form cross-links.

[0233] For example, in the case of a reaction between alkene or alkyne reactive moieties forming a hydrocarbon bridge, a ring-closing metathesis (RCM) reaction may be used. In some embodiments, the RCM reaction involves reacting a peptide in the presence of a ruthenium complex. In some embodiments, the ruthenium complex is, for example, a Grubbs, Grubbs II, Hoveyda-Grubbs I, or Hoveyda-Grubbs II catalyst.

change

[0234] Other examples of metathesis (e.g., RCM) catalysts, reagents, and reaction conditions useful in the present method can be found in, e.g., Schrodi, Y.; Pederson, RL Aldrichimica Acta 2007, 40, 45; Adv. Synth. Catal. 2007, 349, 1-268; Grubbs, RH Tetrahedron 2004, 60, 7117; Handbook of Metathesis; Grubbs, RH, Ed.; Wiley-VCH: Weinheim, 2003; Vols. 1-3; Trnka, TM; Grubbs, RH Acc. Chem. Res. 2001, 34, 18; Furstner, A. Angew. Chem., Int. Ed. 2000, 39, 3012; Schuster, M.; Blechert,S.Angew.Chem.,Int.Ed.1997,36,2036; Ritter,T.et al.Organometallics 2006,25,5740; Chatterjee,AKet al.J.Am.Chem.Soc.2000,122,3783; Chatterjee,AK; Grubbs,RHOrg.Lett.1999,1,1751; Murelli,RP; Snapper,MLOrg.Lett.2007,9,1749; Stewart,ICet al.Org.Lett.2007,9,1589; Ung,T.et al.Organometallics 2004,23,5399; Benitez,D.; Goddard, WA, III. J. Am. Chem. Soc. 2005, 127, 12218; Love, JA et al. Angew. Chem., Int. Ed. 2002, 41, 4035; Sanford, MS et al. Organometallics 2001, 20, 5314; Choi, T.-L.; Grubbs, RH Angew. Chem. 2003, 115, 1785; Ritter, T. et al. Organometallics 2006, 25, 5740, the entire contents of each of which are incorporated herein by reference.

[0235] As another example, in the case of reactions between thiol reactive moieties forming dithio bridges, a thiol-stapling reaction may be used. "Thiol-stapling" involves reacting two thiol moieties in the presence of a cross-linking reagent (e.g., a dihalide) to form a cross-link. Examples of thiol-stapling reactions can be found, for example, in U.S. Patent Application Publication No. US 2019 / 0382443 A1 published December 19, 2019; U.S. Patent No. 9,670,484 B2 issued June 6, 2017; U.S. Patent No. 9,644,201 B2 issued May 9, 2017; U.S. Patent Application Publication No. 2017 / 0067045 A1 published March 9, 2017; Peraro, L. et al. Methods in Enzymology vol. 580 (2016): 303-32; and Kale, S. et al. Nature Chem 10, 715-723 (2018), the entire contents of each of which are incorporated herein by reference.

[0236] For example, provided below in Table 3 are examples of cross-linking reagents and dithio cross-links that are capable of reacting via a thiol-stapling reaction. Table 3. Thiol-stapling cross-linking reagents [Table 3]

[0237] As another example, in the case of a reaction between azide and alkyne reactive moieties forming a triazolylene bridge, an alkyne-azide 1,3-cycloaddition may be used (e.g., Huisgen alkyne-azide cycloaddition). In some embodiments, the alkyne-azide cycloaddition is copper-catalyzed. In some embodiments, the alkyne-azide cycloaddition is strain-promoted. Examples of alkyne-azide reactions can be found, for example, in Kolb, Finn and Sharpless Angewandte Chemie International Edition (2001) 40:2004-2021; Kolb and Sharpless, Drug Discov Today (2003) 24:1128-1137; and Evans, Australian Journal of Chemistry (2007) 60:384-395.

[0238] Other click chemistry reactions may also be used to form crosslinks of the peptides described herein. "Click chemistry" is a chemical approach introduced by Sharpless in 2001, which describes chemistry tailored to rapidly and reliably generate substances by linking small units together. See, for example, Kolb, Finn and Sharpless Angewandte Chemie International Edition (2001) 40: 2004-2021; Evans, Australian Journal of Chemistry (2007) 60: 384-395. Exemplary coupling reactions (some of which may be classified as "click chemistry") include, but are not limited to, the formation of esters, thioesters, and amides from activated acids or acyl halides (e.g., peptide coupling); nucleophilic substitution reactions (e.g., nucleophilic displacement of halides or ring opening of strained ring systems); azide-alkyne Huisgen cycloadditions; thiol-yne additions; imine formation; Michael additions (e.g., maleimide additions); and Diels-Alder reactions (e.g., tetrazine [4+2] cycloadditions).

[0239] Pharmaceutical Compositions, Kits, and Administration The present disclosure provides a pharmaceutical composition comprising the peptide disclosed herein or a pharma- ceutically acceptable salt thereof. The pharmaceutical composition may comprise one or more pharma- ceutically acceptable carriers / excipients. In some embodiments, the peptide described herein is provided in an effective amount in the pharmaceutical composition. In some embodiments, the effective amount is an amount that is therapeutically effective (e.g., for treating a bacterial infection in a subject). In some embodiments, the effective amount is an amount that is prophylactically effective (e.g., for preventing a bacterial infection in a subject). In some embodiments, the effective amount is an amount that is effective to kill and / or inhibit the growth of bacteria in a subject or biological sample.

[0240] The pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology.In general, such preparation methods include bringing into association the peptides described herein (i.e., "active ingredient") with a carrier or excipient, and / or one or more other accessory ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into a desired single-dose or multi-dose unit.

[0241] Pharmaceutical compositions can be prepared, packaged, and / or sold in bulk, as single unit doses, and / or as a plurality of single unit doses. A "unit dose" is a discrete amount of pharmaceutical composition that contains a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject, and / or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

[0242] The relative amounts of the active ingredient, pharma- ceutically acceptable excipient, and / or any additional ingredients in the pharmaceutical compositions described herein may vary depending on the identity, size, and / or condition of the subject being treated, and further depending on the route by which the composition is to be administered. The compositions may contain between 0.1% and 100% (w / w) of the active ingredient.

[0243] Pharmaceutically acceptable excipients used in the manufacture of the provided pharmaceutical compositions include inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrating agents, binders, preservatives, buffering agents, lubricating agents, and / or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening agents, flavoring agents, and perfuming agents may also be present in the composition.

[0244] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and mixtures thereof.

[0245] Exemplary granulating and / or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0246] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, ... ), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween® 20), polyoxyethylene sorbitan (Tween® 60), polyoxyethylene sorbitan monooleate (Tween® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan tristearate (Span® 65), glyceryl monooleate, sorbitan monooleate (Span® 80)) , polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj® 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol®), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether (Brij® 30)), poly(vinylpyrrolidone), diethylene glycol monolaurate,Triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic® F-68, Poloxamer® P-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and / or mixtures thereof.

[0247] Exemplary binders include starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, and the like), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate (Veegum®), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and / or mixtures thereof.

[0248] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoan preservatives, alcohol preservatives, acidic preservatives, and other preservatives. In some embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0249] Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0250] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and its salts and hydrates (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, etc.), citric acid and its salts and hydrates (e.g., citric acid monohydrate), fumaric acid and its salts and hydrates, malate and its salts and hydrates, phosphoric acid and its salts and hydrates, and tartaric acid and its salts and hydrates. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0251] Exemplary antifungal preservatives include butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0252] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.

[0253] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0254] Other preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant® Plus, Phenonip®, methylparaben, Germall® 115, Germaben® II, Neolone®, Kathon®, and Euxyl®.

[0255] Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and mixtures thereof.

[0256] Exemplary lubricants include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0257] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, chamomile, canola, caraway, carnauba, castor oil, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, emu, eucalyptus, evening primrose, fish, linseed, geraniol, loofah, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon. Examples of synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecyl oleate, oleyl alcohol, silicone oil, and mixtures thereof. In some embodiments, the formulation includes a polymeric excipient. In some embodiments, the formulation includes a polyether. In some embodiments, the formulation comprises polyethylene glycol (PEG) (e.g., PEG200, PEG300, PEG400, etc.).

[0258] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to active ingredients, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (such as cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.In addition to inert diluents, oral compositions may contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and aromatics. In certain embodiments for parenteral administration, the conjugates described herein are mixed with a solubilizing agent such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

[0259] In the injectable preparation, for example, sterile injectable aqueous or oily suspension can be formulated according to known technology using suitable dispersing or wetting agent and suspending agent. Sterile injectable preparation can be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution (USP) and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspending medium. For this purpose, any non-irritating fixed oil can be used, including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the injectable preparation.

[0260] The injectable preparations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0261] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharma- ceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or with (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants, such as glycerol, (d) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarders, such as paraffin. (f) absorption enhancers such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include buffering agents.

[0262] Solid compositions of a similar type may be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols, etc. The solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells, such as enteric coatings and other coatings known in the art of pharmacology. They may optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of encapsulating compositions that may be used include polymeric substances and waxes. Solid compositions of a similar type may be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols, etc.

[0263] The active ingredient may be in microencapsulated form with one or more excipients as mentioned above. The solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells, such as enteric coatings, release controlling coatings, and other coatings known in the pharmaceutical formulating art. In such solid dosage forms, the active ingredient may be admixed with at least one inert diluent, such as sucrose, lactose, or starch. Such solid dosage forms may contain additional substances other than the inert diluents, as is normal practice, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also contain buffering agents. They may optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of encapsulating compositions that may be used include polymeric substances and waxes.

[0264] Dosage forms for topical and / or transdermal administration of the peptides described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and / or patches. In general, the active ingredient is mixed under sterile conditions with a pharma- ceutically acceptable carrier or excipient, and / or any preservatives and / or buffers as required. In addition, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of the active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and / or dispensing the active ingredient in a proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate-controlling membrane and / or dispersing the active ingredient in a polymer matrix and / or gel.

[0265] Suitable devices for use in the delivery of the intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices that limit the effective penetration length of the needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical Mantoux method of intradermal administration. Jet injection devices that deliver liquid vaccines to the dermis via a liquid jet injection device and / or via a needle that creates a jet that penetrates the stratum corneum and reaches the dermis are suitable. Ballistic powder / particle delivery devices that use compressed gas to accelerate the peptide in powder form through the outer layer of the skin into the dermis are suitable.

[0266] Formulations suitable for topical administration include, but are not limited to, liquid and / or semi-liquid preparations such as liniments, lotions, oil-in-water and / or water-in-oil emulsions such as creams, ointments, and / or pastes, and / or solutions and / or suspensions. Topically administrable formulations may contain, for example, from about 1% to about 10% (w / w) of the active ingredient, although the concentration of the active ingredient is increased up to the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further include one or more additional ingredients as described herein.

[0267] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing a conjugate described herein with a suitable nonirritating excipient or carrier such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active ingredient.

[0268] The pharmaceutical compositions described herein may be prepared, packaged, and / or sold in a formulation suitable for pulmonary administration via the oral cavity. Such formulations may include dry particles containing the active ingredient. The pharmaceutical compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and / or suspension. Such formulations may be prepared, packaged, and / or sold as aqueous and / or dilute alcoholic solutions and / or suspensions containing the active ingredient, optionally sterilized, and may be conveniently administered using either a nebulizer and / or atomizer device.

[0269] The formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of the pharmaceutical compositions described herein. Formulations for nasal administration may, for example, contain as little as about 0.1% (w / w) to as much as 100% (w / w) of the active ingredient, and may contain one or more additional ingredients described herein.

[0270] The pharmaceutical compositions described herein may be prepared, packaged, and / or sold in a formulation for buccal administration. Such formulations may be, for example, in the form of tablets, and / or lozenges made using conventional methods, and may contain, for example, 0.1-20% (w / w) of the active ingredient, the remainder comprising an orally dissolvable and / or degradable composition, and optionally one or more of the additional ingredients described herein. Alternatively, a formulation for buccal administration may comprise a powder and / or an aerosolized and / or atomized solution and / or suspension comprising the active ingredient.

[0271] The pharmaceutical compositions described herein may be prepared, packaged, and / or sold in a formulation for administration to the eye. Such formulations may be in the form of, for example, eye drops comprising, for example, a 0.1 to 1.0% (w / w) solution and / or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops may further comprise buffering agents, salts, and / or one or more other of the additional ingredients described herein. Other useful ophthalmically administrable formulations include those comprising the active ingredient in microcrystalline form and / or in a liposomal preparation. Ear drops and / or eye drops are also contemplated as being within the scope of this disclosure.

[0272] Although the description of pharmaceutical compositions provided herein is primarily directed to pharmaceutical compositions suitable for administration to humans, it will be understood by those skilled in the art that such compositions are generally suitable for administration to any and all animals. Modifications of pharmaceutical compositions suitable for administration to humans to make compositions suitable for administration to a variety of animals are well understood, and the ordinarily skilled veterinary pharmacologist may design and / or perform such modifications using routine experimentation.

[0273] The peptides provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage.However, it will be understood that the total daily usage of the compositions described herein will be determined by a physician within the scope of sound medical judgment.The specific therapeutically effective dose level for any specific subject or organism will depend on a variety of factors, including the severity of the disease and disorder being treated; the activity of the specific active ingredient being adopted; the specific composition being adopted; the age, weight, general health, sex and dietary habits of the subject; the time of administration, route of administration and the excretion rate of the specific active ingredient being adopted; the duration of treatment; the drugs that are used in combination with or simultaneously with the specific active ingredient being adopted; and the like factors well known in the medical field.

[0274] The peptides and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subcutaneous, intracerebroventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and / or drops), mucosal, nasal, buccal, sublingual; by intratracheal instillation, bronchial instillation, and / or inhalation; and / or as oral spray, nasal spray, and / or aerosol. Particularly contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), local administration via blood and / or lymphatic supply, topical administration, and / or direct administration to the affected area. In general, the most appropriate route of administration will depend on a variety of factors, including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration).

[0275] The exact amount of peptide required to achieve an effective dose will vary from subject to subject, depending, for example, on the species, age, and general condition of the subject, the severity of side effects or disorders, the identity of the specific peptide, the mode of administration, etc. An effective amount may be included in a single dose (e.g., a single oral dose) or multiple doses (e.g., multiple oral doses). In some embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, any two doses of the multiple doses include different or substantially the same amount of the peptide described herein. In some embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the frequency of administering the multiple doses to a subject or applying the multiple doses to a tissue or cell is 3 doses per day, 2 doses per day, 1 dose per day, 1 dose every other day, 1 dose every 2 days, 1 dose every week, 1 dose every 2 weeks, 1 dose every 3 weeks, or 1 dose every 4 weeks. In some embodiments, when multiple doses are administered to a subject or applied to tissue or cell, the period between the first and last doses of the multiple doses is 1 day, 2 days, 4 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 6 months, 9 months, 1 year, 2 years, 3 years, 4 years, 5 years, 7 years, 10 years, 15 years, 20 years, or the lifespan of the subject, tissue, or cell.In some embodiments, the period between the first and last doses of the multiple doses is 3 months, 6 months, or 1 year.In some embodiments, the period between the first and last doses of the multiple doses is the lifespan of the subject, tissue, or cell.

[0276] In certain embodiments, the doses described herein (e.g., either a single dose, or multiple doses) independently include between 0.1 μg and 1 μg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g (inclusive) of a peptide described herein.

[0277] The dosage ranges as described herein provide guidance for administration of the provided pharmaceutical compositions to adults. For example, the amount to be administered to a child or adolescent can be determined by a physician or person skilled in the art and can be less than or the same as that administered to an adult.

[0278] The peptide or composition may be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and / or prophylactically active agents) as described herein. The peptide or composition may be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., efficacy and / or effectiveness) in treating a disease in a subject in need thereof, preventing a disease in a subject in need thereof, reducing the risk of developing a disease in a subject in need thereof), improve bioavailability, improve safety, reduce drug resistance, reduce and / or modify metabolism, inhibit excretion, and / or modify distribution in a subject or cell. It will also be understood that the treatments employed may achieve a desired effect on the same disorder and / or may achieve different effects. In some embodiments, the pharmaceutical compositions described herein that include the peptides described herein and the additional pharmaceutical agents exhibit a synergistic effect that is not present in pharmaceutical compositions that include one of the peptides and the additional pharmaceutical agents but not both. In some embodiments, the additional pharmaceutical agents achieve a desired effect on the same disorder. In some embodiments, the additional pharmaceutical agents achieve different effects.

[0279] The peptide or composition may be administered simultaneously with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful, for example, as a combination therapy. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules, such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration, as provided in the U.S. Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic peptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNA, RNA, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells.

[0280] Additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-angiogenic agents, steroidal or non-steroidal anti-inflammatory agents (NSAIDs), immunosuppressants, antibacterial agents, antiviral agents, cardiovascular agents, cholesterol lowering agents, antidiabetic agents, anti-allergy agents, contraceptives, pain-relieving agents, anesthetics, anticoagulants, enzyme inhibitors, steroidal agents, steroidal or antihistamines, antigens, vaccines, antibodies, decongestants, sedatives, opioids, analgesics, antipyretics, hormones, and prostaglandins.

[0281] In some embodiments, the additional pharmaceutical agent is an antimicrobial agent. In some embodiments, the additional pharmaceutical agent is an antibiotic. In some embodiments, the additional pharmaceutical agent is selected from the group consisting of polymyxin, rifampicin, ofloxacin, vancomycin, and meropenem.

[0282] Each additional pharmaceutical agent may be administered at a dose and / or schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and / or with the peptides or compositions described herein in a single dose or composition, or may be administered separately in different doses or compositions. The particular combination employed in the regimen will take into consideration the compatibility of the peptides described herein with the additional pharmaceutical agent(s) and / or the desired therapeutic and / or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agent(s) in the combination will be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in the combination will be lower than the levels utilized individually.

[0283] Also covered by the present disclosure are kits (e.g., pharmaceutical packs). The kits provided may include a pharmaceutical composition or peptide described herein and a container (e.g., a vial, an ampoule, a bottle, a syringe, and / or a dispenser package, or other suitable container). In some embodiments, the kits provided may further include a second container, optionally including a pharmaceutical excipient for diluting or suspending the pharmaceutical composition or peptide described herein. In some embodiments, the pharmaceutical composition or peptide provided in the first container and the second container described herein are combined to form one unit dosage form. Thus, in one aspect, provided is a kit including a first container including a peptide or pharmaceutical composition described herein. In some embodiments, the kit is useful for treating a disease (e.g., a bacterial infection) in a subject in need thereof. In some embodiments, the kit is useful for preventing a disease (e.g., a bacterial infection) in a subject in need thereof.

[0284] In some embodiments, the kit described herein further comprises instructions for using the kit. The kit described herein may also comprise information as required by regulatory authorities such as the U.S. Food and Drug Administration (FDA). In some embodiments, the information contained in the kit is prescription information. In some embodiments, the kit provides instructions for treating a disease (e.g., bacterial infection) in a subject in need thereof. In some embodiments, the kit provides instructions for preventing a disease (e.g., bacterial infection) in a subject in need thereof. The kit described herein may comprise one or more additional pharmaceutical agents as separate compositions as described herein.

[0285] Lipid formulations Also provided herein are formulations (i.e., pharmaceutical compositions) comprising one or more stapled antimicrobial peptides (StAMPs) and one or more lipids ("lipid formulations"). The lipid formulation may be a liposomal formulation, a micellar formulation, a lipid nanoparticle ("LNP") formulation, or the like. In some embodiments, the lipid formulation is a micellar formulation. In some embodiments, the lipid formulation is a liposomal formulation. In some embodiments, the lipid formulation is a LNP formulation. In some embodiments, the StAMP is encapsulated in a micelle, liposome, LNP, or the like.

[0286] In some embodiments, the lipid formulation comprises a peptide (for example, StAMP) provided herein. In some embodiments, the peptide (for example, StAMP) is a peptide described, for example, in International PCT Application Publication No. WO 2017 / 004591 published on January 5, 2017; and WO 2019 / 018499 published on January 24, 2019 (the entire contents of each of which are incorporated herein by reference).

[0287] The lipid formulations provided herein include one or more lipids. In some embodiments, the lipid includes a phospholipid (e.g., 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)). In some embodiments, the lipid includes polyethylene glycol (PEG). In some embodiments, the lipid is a PEGylated phospholipid. A "PEGylated phospholipid" is a lipid that includes a phospholipid conjugated to PEG. For example, in some embodiments, the PEGylated phospholipid includes DSPE and PEG (i.e., "DSPE-PEG"). In some embodiments, the PEGylated phospholipid is a DSPE-MPEG (e.g., DSPE-MPEG(2000)). In some embodiments, the PEGylated phospholipid is a DSPE-PEG amine (e.g., DSPE-PEG(2000) amine). The lipid (e.g., a phospholipid) includes a pharma- ceutically acceptable salt of the lipid. For example, the structure of DSPE-MPEG is shown below: [ka]

[0288] For example, the structure of DSPE-PEG amine is shown below: [ka]

[0289] The lipid formulation may contain any peptide:lipid ratio. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:1 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:2 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:2.5 to about 1:20 (w / w), inclusive. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:2 to about 1:3 (w / w), inclusive. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:4 to about 1:6 (w / w), inclusive. In some embodiments, the lipid formulation contains a peptide:lipid ratio of about 1:8 to about 1:12 (w / w), inclusive. In certain embodiments, the lipid formulation comprises a peptide:lipid ratio of about 1:15 to about 1:25 (w / w), inclusive.

[0290] In some embodiments, the lipid formulation comprises a peptide:lipid ratio of about 1:2.5 (w / w). In some embodiments, the lipid formulation comprises a peptide:lipid ratio of about 1:5 (w / w). In some embodiments, the lipid formulation comprises a peptide:lipid ratio of about 1:10 (w / w). In some embodiments, the lipid formulation comprises a peptide:lipid ratio of about 1:20 (w / w).

[0291] For example, in some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:1 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:2 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:2.5 to about 1:20 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:2 to about 1:3 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:4 to about 1:6 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:8 to about 1:12 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:15 to about 1:25 (w / w), inclusive.

[0292] In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:2.5 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:5 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:10 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-MPEG ratio of about 1:20 (w / w).

[0293] For example, in some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:1 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:2 to about 1:25 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:2.5 to about 1:20 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:2 to about 1:3 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:4 to about 1:6 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:8 to about 1:12 (w / w), inclusive. In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:15 to about 1:25 (w / w), inclusive.

[0294] In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:2.5 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:5 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:10 (w / w). In some embodiments, the lipid formulation comprises a peptide:DSPE-PEG amine ratio of about 1:20 (w / w).

[0295] Treatment Methods and Uses The peptides of the present disclosure (e.g., stapled peptides, e.g., StAMP) have antimicrobial activity and are therefore useful in a variety of methods, such as treating infectious diseases (e.g., bacterial infections) in a subject, and killing and / or inhibiting the growth of microorganisms (e.g., bacteria).

[0296] Provided herein is a method for treating and / or preventing an infectious disease in a subject, comprising administering to the subject a peptide described herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the infectious disease is a bacterial infection. In some embodiments, the infectious disease is a viral infection. In some embodiments, the infectious disease is a protozoan infection. In some embodiments, the infectious disease is a fungal infection. In some embodiments, the infectious disease is a parasitic infection.

[0297] In some embodiments, the method is for treating an infectious disease. In some embodiments, the method is for preventing an infectious disease.

[0298] Provided herein is a method for treating and / or preventing bacterial infection in a subject, comprising administering to the subject a peptide described herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the bacterial infection is a gram-positive bacterial infection, i.e., caused by gram-positive bacteria. In some embodiments, the bacterial infection is a gram-negative bacterial infection, i.e., caused by gram-negative bacteria.

[0299] In some embodiments, the method is for treating a bacterial infection. In some embodiments, the method is for preventing a bacterial infection.

[0300] The gram-negative bacterial infection may be caused by any of the gram-negative bacteria described herein. In some embodiments, the bacterial infection is caused by Acineobacter, Escherichia, Pseudomonas, Neisseria, Chlamydia, Yersinia, Proteus, Enterobacter, Serratia, Helicobacter, Salmonella, Shigella, Moraxella, Stenotrophomonas, Bdellovibrio, Klebsiella, Legionella, or Acetobacter. In some embodiments, the bacterial infection is caused by Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Neisseria gonorrhoeae, Chlamydia trachomatis, Yersinia pestis, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, Helicobacter pylori, Salmonella enteritidis, Salmonella typhi, Shigella, Klebsiella pneumoniae, or Legionella pneumophila bacteria, or a gram-negative bacterium containing the MCR-1 gene.

[0301] In some embodiments, the bacterial infection is caused by a species of Escherichia, Acinetobacter, Pseudomonas, or Klebsiella. In some embodiments, the bacterial infection is caused by Escherichia coli (E. coli), Acinetobacter baumannii (A. baumannii), Pseudomonas aeruginosa (P. aeruginosa), or Klebsiella pneumoniae (K. pneumoniae). In some embodiments, the bacterial infection is caused by a species of Stenotrophomonas, Burkholderia, or Klebsiella. In some embodiments, the bacterial infection is caused by Stenotrophomonas maltophilia (S. maltophilia), Burkholderia cepacia (B. cepacia), or Klebsiella oxytoca (K. oxytoca). In some embodiments, the bacterial infection is caused by E. coli. In some embodiments, the bacterial infection is caused by A. baumannii. In some embodiments, the bacterial infection is caused by P. aeruginosa. In some embodiments, the bacterial infection is caused by K. pneumoniae. In some embodiments, the bacterial infection is caused by S. maltophilia. In some embodiments, the bacterial infection is caused by B. cepacia. In some embodiments, the bacterial infection is caused by K. oxytoca.

[0302] In some embodiments, the bacterial infection is caused by a species of Enterobacter, Pasteurella, Proteus, or Citrobacter. Examples of species are provided herein.

[0303] In some embodiments, the bacterial infection is an antibiotic-resistant bacterial infection, i.e., a bacterial infection caused by antibiotic-resistant bacteria. In some embodiments, the bacterial infection is a gram-negative, antibiotic-resistant bacterial infection. Thus, also provided herein is a method for treating an antibiotic-resistant gram-negative bacterial infection in a subject, the method comprising administering to the subject a peptide described herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the antibiotic-resistant bacterial infection is caused by bacteria resistant to one or more antibiotics selected from the group consisting of polymyxin, carbapenem, aminoglycoside, vancomycin, methicillin, clarithromycin, cephalosporin, penicillin, ampicillin, fluoroquinolone, tetracycline, ceftazidime, ceftriaxone, ciprofloxacin, gentamicin, tobramycin, and β-lactam.

[0304] As discussed herein, the present disclosure relates in part to antimicrobial peptides that have reduced toxicity compared to known or existing antimicrobial agents (e.g., AMPs, e.g., StAMP). Reduced toxicity can be, but is not limited to, reduced nephrotoxicity, reduced hepatotoxicity, and / or reduced hemolytic activity.

[0305] In some embodiments, the peptides provided herein (e.g., StAMP) have reduced nephrotoxicity compared to a reference. Nephrotoxicity is reduced if the peptide kills and / or damages fewer kidney cells compared to a reference. In some embodiments, the peptides have reduced nephrotoxicity compared to the corresponding peptides comprising SEQ ID NO: 1 or 2. In some embodiments, the peptides have reduced nephrotoxicity compared to pexiganan. In some embodiments, the peptides have reduced nephrotoxicity compared to magainin. In some embodiments, the peptides have reduced nephrotoxicity compared to magainin II.

[0306] In some embodiments, the peptides provided herein have a nephrotoxicity that is at least 10% (e.g., at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%) lower than that of the reference. The nephrotoxicity of the peptides can be measured in vitro using a renal proximal tubule epithelial cell (RPTEC) nephrotoxicity assay.

[0307] In some embodiments, the peptides provided herein (e.g., StAMP) have reduced hepatotoxicity compared to a reference. Hepatotoxicity is reduced if the peptide kills and / or damages fewer liver cells compared to a reference. In some embodiments, the peptides have reduced hepatotoxicity compared to the corresponding peptides comprising SEQ ID NO: 1 or 2. In some embodiments, the peptides have reduced hepatotoxicity compared to pexiganan. In some embodiments, the peptides have reduced hepatotoxicity compared to magainin. In some embodiments, the peptides have reduced hepatotoxicity compared to magainin II.

[0308] In some embodiments, the peptides provided herein have a hepatotoxicity that is at least 10% (e.g., at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%) lower than that of the reference. The hepatotoxicity of the peptides can be measured in vitro using a human liver cell (e.g., HepG2) toxicity assay.

[0309] In some embodiments, the peptides provided herein (e.g., StAMP) have reduced hemolytic activity compared to a reference. "Hemolytic activity" for purposes of this disclosure refers to the destruction or lysis of red blood cells. In some embodiments, the peptides have reduced hemolytic activity compared to the corresponding peptides comprising SEQ ID NO: 1 or 2. In some embodiments, the peptides have reduced hemolytic activity compared to melittin. In some embodiments, the peptides have reduced hemolytic activity compared to magainin. In some embodiments, the peptides have reduced hemolytic activity compared to magainin II.

[0310] In some embodiments, the peptides provided herein have a hemolytic activity that is at least 10% (e.g., at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%) lower than the hemolytic activity of the reference.

[0311] Also provided herein are methods of killing and / or inhibiting the growth of bacteria (e.g., gram-negative and / or gram-positive bacteria), the methods comprising contacting the bacteria (e.g., in vitro or in vivo) with a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0312] In some embodiments, the bacteria is a gram-positive bacterium. In some embodiments, the bacteria is a gram-negative bacterium. In some embodiments, the gram-negative bacterium is Acineobacter, Escherichia, Pseudomonas, Neisseria, Chlamydia, Yersinia, Proteus, Enterobacter, Serratia, Helicobacter, Salmonella, Shigella, Moraxella, Stenotrophomonas, Bdellovibrio, Klebsiella, Legionella, or Acetobacter. In some embodiments, the bacterium is an Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Neisseria gonorrhoeae, Chlamydia trachomatis, Yersinia pestis, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, Helicobacter pylori, Salmonella enteritidis, Salmonella typhi, Shigella, Klebsiella pneumoniae, or Legionella pneumophila bacterium, or a Gram-negative bacterium containing the MCR-1 plasmid.

[0313] In some embodiments, the bacterium is a species of Escherichia, Acinetobacter, Pseudomonas aeruginosa, or Klebsiella. In some embodiments, the bacterium is Escherichia coli (E. coli), Acinetobacter baumannii (A. baumannii), Pseudomonas aeruginosa (P. aeruginosa), or Klebsiella pneumoniae (K. pneumoniae). In some embodiments, the bacterium is a species of Stenotrophomonas, Burkholderia, or Klebsiella. In some embodiments, the bacterium is Stenotrophomonas maltophilia (S. maltophilia), Burkholderia cepacia (B. cepacia), or Klebsiella oxytoca (K. oxytoca). In some embodiments, the bacterium is E. coli. In some embodiments, the bacterium is A. baumannii. In some embodiments, the bacterium is P. aeruginosa. In some embodiments, the bacterium is K. pneumoniae. In some embodiments, the bacterium is S. maltophilia. In some embodiments, the bacterium is B. cepacia. In some embodiments, the bacterium is K. oxytoca.

[0314] In some embodiments, the bacterium is a species of Enterobacter, Pasteurella, Proteus, or Citrobacter. Examples of species are provided herein.

[0315] In some embodiments, the bacteria is an antibiotic-resistant bacteria. In some embodiments, the bacteria is a gram-negative, antibiotic-resistant bacteria. Thus, also provided herein is a method of killing and / or inhibiting the growth of antibiotic-resistant, gram-negative bacteria with a peptide described herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof (e.g., in vitro or in vivo). In some embodiments, the bacteria is resistant to one or more antibiotics selected from the group consisting of polymyxins, carbapenems, aminoglycosides, vancomycin, methicillin, clarithromycin, cephalosporins, penicillins, ampicillins, fluoroquinolones, tetracyclines, ceftazidime, ceftriaxone, ciprofloxacin, gentamicin, tobramycin, and β-lactams.

[0316] In some embodiments, the peptide (e.g., StAMP) has a minimum inhibitory concentration (MIC) of less than about 64 μg / mL. In some embodiments, the peptide (e.g., StAMP) has a minimum inhibitory concentration (MIC) of less than or equal to about 32 μg / mL, less than or equal to about 16 μg / mL, less than or equal to about 14 μg / mL, less than or equal to about 12 μg / mL, or less than or equal to about 10 μg / mL. In some embodiments, the peptide (e.g., StAMP) has a minimum inhibitory concentration (MIC) of less than or equal to about 10 μg / mL. In some embodiments, the peptide (for example, StAMP) has a MIC of less than or equal to about 10 μg / mL, less than or equal to about 9 μg / mL, less than or equal to about 8 μg / mL, less than or equal to about 7 μg / mL, less than or equal to about 6 μg / mL, less than or equal to about 5 μg / mL, less than or equal to about 4 μg / mL, less than or equal to about 3 μg / mL, less than or equal to about 2 μg / mL, or less than or equal to about 1 μg / mL. In some embodiments, the peptide (for example, StAMP) has a MIC of about 2 μg / mL. In some embodiments, the MIC is at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 1.9 μg / mL.

[0317] In some embodiments, the peptide (e.g., StAMP) has an MIC of about 0.1 to about 64 μg / mL, about 0.1 to about 32 μg / mL, about 0.1 to about 16 μg / mL, about 0.1 to about 14 μg / mL, about 0.1 to about 12 μg / mL, or about 0.1 to about 10 μg / mL. In some embodiments, the peptide (e.g., StAMP) has an MIC of about 0.1 to about 10 μg / mL. In some embodiments, the peptide (e.g., StAMP) has an MIC of about 0.1 to about 9 μg / mL, about 0.1 to about 8 μg / mL, about 0.1 to about 7 μg / mL, about 0.1 to about 6 μg / mL, about 0.1 to about 5 μg / mL, about 0.1 to about 4 μg / mL, about 0.1 to about 3 μg / mL, or about 0.1 to about 2 μg / mL. In some embodiments, a peptide (e.g., StAMP) has a MIC of about 0.1 to about 4 μg / mL. In some embodiments, the MIC is at least about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 1.9 μg / mL.

[0318] In some embodiments, the peptides described herein (e.g., StAMP) selectively kill and / or inhibit the growth of Gram-negative bacteria over Gram-positive bacteria, hi some embodiments, the peptides selectively kill and / or inhibit the growth of Escherichia coli (E. coli), Acinetobacter baumannii (A. baumannii), Pseudomonas aeruginosa (P. aeruginosa), and / or Klebsiella pneumoniae (K. pneumoniae) over Staphylococcus aureus (S. aureus).

[0319] In some embodiments, the peptides provided herein (e.g., StAMP) have the ability to kill and / or inhibit the growth of microbial cells but not mammalian cells. In some embodiments, the peptides provided herein (e.g., StAMP) have the ability to selectively lyse microbial cells (e.g., bacterial cells) over mammalian cells. In some embodiments, the peptides (e.g., StAMP) have selective cytotoxicity against microbial cells (e.g., bacterial cells) over mammalian cells. This selectivity can lead to reduced toxicity (e.g., reduced nephrotoxicity, hepatotoxicity, hemolytic activity) when the peptides are administered to a subject.

[0320] Provided herein is a method for selectively killing and / or inhibiting the growth of microbial cells (e.g., bacterial cells) over mammalian cells, the method comprising contacting microbial (e.g., bacterial) cells and mammalian cells with a peptide (e.g., StAMP) provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, a peptide (e.g., StAMP) provided herein lyses microbial cells (e.g., bacterial cells) to a greater extent than mammalian cells. Thus, provided herein is a method for selectively lysing microbial cells (e.g., bacterial cells) over mammalian cells, the method comprising contacting microbial (e.g., bacterial) cells and mammalian cells with a peptide (e.g., StAMP) provided herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0321] In some embodiments, the microbial cell is a bacterial cell. In some embodiments, the microbial cell is a gram-negative bacterial cell. The cell may comprise any of the bacteria (e.g., gram-negative bacteria) provided herein. In some embodiments, the bacterial cell is an Escherichia, Acinetobacter, Pseudomonas aeruginosa, or Klebsiella cell. In some embodiments, the bacterial cell is an Escherichia coli (E. coli), Acinetobacter baumannii (A. baumannii), Pseudomonas aeruginosa (P. aeruginosa), or Klebsiella pneumoniae (K. pneumoniae) cell. In some embodiments, the bacterial cell is a Stenotrophomonas, Burkholderia, or Klebsiella cell. In some embodiments, the bacterial cell is a Stenotrophomonas maltophilia (S. maltophilia), Burkholderia cepacia (B. cepacia), or Klebsiella oxytoca (K. oxytoca) cell. In some embodiments, the bacterial cell is an E. coli cell. In some embodiments, the bacterial cell is an A. baumannii cell. In some embodiments, the bacterial cell is a P. aeruginosa cell. In some embodiments, the bacterial cell is a K. pneumoniae cell. In some embodiments, the bacterial cell is a S. maltophilia cell. In some embodiments, the bacterial cell is a B. cepacia cell. In some embodiments, the bacterial cell is a K. oxytoca cell.

[0322] In some embodiments, the bacterial cell is an Enterobacter, Pasteurella, Proteus, or Citrobacter cell. Exemplary species are provided herein.

[0323] Also provided herein are peptides described herein (e.g., StAMP), and pharma- ceutically acceptable salts thereof, and pharmaceutical compositions thereof, for use in any of the methods described herein (e.g., for treating an infectious disease in a subject, for treating a bacterial infection in a subject, for killing and / or inhibiting the growth of bacteria in a subject, for selectively killing and / or inhibiting the growth of bacterial cells over mammalian cells in a subject, etc.).

[0324] In another aspect, also provided herein is the use of the peptides described herein (e.g., StAMP), and pharma- ceutically acceptable salts thereof, and pharmaceutical compositions thereof, for the manufacture of a medicament (e.g., for treating an infectious disease in a subject, for treating a bacterial infection in a subject, for killing and / or inhibiting the growth of bacteria in a subject, for selectively killing and / or inhibiting the growth of bacterial cells over mammalian cells in a subject, etc.).

[0325] In some embodiments, the in vivo (i.e., in a subject) methods and uses provided herein include administering to a subject an effective amount of a peptide provided herein (e.g., StAMP), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the effective amount is a therapeutically effective amount. In some embodiments, the effective amount is a prophylactically effective amount.

[0326] In some embodiments, the in vitro methods and uses provided herein can be carried out, for example, in cell assays or biological samples. In some embodiments, the in vitro methods and uses include contacting microbial cells (for example, bacterial cells) with the peptides provided herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions thereof.

[0327] example Common methods Solid Phase Peptide Synthesis: Antimicrobial peptides and their stapled derivatives were synthesized using Fmoc-based solid-phase peptide synthesis. To achieve i+4 staple length, α-methyl, α-alkenyl amino acids were used for flanking three residues. For the stapling reaction, Grubbs first generation ruthenium catalyst dissolved in dichloroethane was added to the peptide while still on the resin. Three to five rounds of stapling were performed to ensure maximum conversion. Once stapled, the peptides were cleaved from the resin using trifluoroacetic acid and then precipitated using a hexane:ether (1:1) mixture, followed by purification using preparative HPLC. Final peptide characterization for purity was assessed using a UHPLC / MS system.

[0328] Antimicrobial activity assay: The following microbroth dilution protocol was adapted from that of the CLSI to determine the minimum inhibitory concentration (MIC) of StAMP. Briefly, Mueller-Hinton Broth (MHB) or cation-adjusted Mueller-Hinton Broth (MHB II) was prepared and autoclaved. Colonies of bacterial cells were then picked and grown in broth overnight at 37°C, then diluted and grown again for 3-4 hours. Serial dilutions of peptide stocks in water (10 μl) were prepared using clear round-bottom polypropylene 96-well plates. 5x10 5 90 μl of bacteria in broth was added to give a final inoculum of CFU / ml. Plates were then covered with porous tape to reduce evaporation and incubated at 37°C for 20-24 hours. The MIC was the lowest peptide concentration at which no visible bacterial growth was observed.

[0329] Cell culture: RPTEC were maintained in MEM medium supplemented with growth factors. HEPG2 cell line was maintained in Eagle's Minimum Essential Medium (EMEM) supplemented with fetal bovine serum to a final concentration of 10%.

[0330] 90 min cytotoxicity assay: Cells were seeded in a 96-well format and after overnight incubation, the medium was replaced with fresh medium. Serial dilutions of StAMP from a 5 mg / mL water stock or vehicle were added to the cells in a final volume of 100 μl. After 90 min incubation at 37° C., 50 μl of cell culture medium was transferred to a clear 96-well plate and incubated with 50 μl of lactate dehydrogenase (LDH) assay reagent for 10 min, and absorbance was measured at 490 nm on a microplate reader.

[0331] 48 hour cytotoxicity assay: Cells were seeded in a 96-well format and incubated for 4 hours to allow attachment, then serial dilutions of StAMP from a 5 mg / mL water stock, or vehicle, were added to the cells in a final volume of 100 μl. After 48 hours of incubation at 37° C., 100 μl of CellTiter-Glo® reagent was added to the cells and the plate was incubated at room temperature for 10 minutes. Luminescence was then measured on a microplate reader.

[0332] Antimicrobial activity and toxicity As shown in Tables A-D, the antimicrobial peptides provided herein (e.g., StAMP) exhibit broad Gram-negative antimicrobial activity and are selective for bacterial cells over kidney and liver cells. Nephrotoxicity was measured after 90 minutes of incubation, while hepatotoxicity was measured after 48 hours of incubation with StAMP. N / =not tested. The following applies to all peptides listed in Tables A-C (i.e., StAMP): Cross-linking (X 1 -X 2 and X 3 -X 4 ):(alk); N-terminus: -NH2; C-terminus: amidated with -NH2; counter ion: Cl - The following applies to all peptides listed in Table D (i.e., StAMP): Crosslink (X 1 -X 2 and X 3 -X 4):(alk); N-terminus: -NH2; C-terminus: as shown in the table; counter ion: Cl - . Table A. Antimicrobial Activity and Nephro / Hepatotoxicity [Table A-1] [Table A-2] [Table A-3] Table B. Antimicrobial Activity and Nephro / Hepatotoxicity [Table B-1] [Table B-2] Table C. Antimicrobial Activity and Nephro / Hepatotoxicity [Table C] Table D. Antimicrobial Activity and Nephro / Hepatotoxicity [Table D]

[0333] Lipid formulations StAMP micelle formulation: DSPE-MPEG(2000) was suspended overnight at 50-100 mg / mL in buffered saline. A StAMP stock solution at 5-20 mg / mL in buffered saline was prepared and equal volumes of micelles and StAMP stock solution were mixed at the desired StAMP:lipid ratio. The suspension was allowed to stand at room temperature for 5-10 minutes before use in in vitro and in vivo assays.

[0334] Pharmacokinetic profile in mice: StAMP solution or micelle suspension was injected subcutaneously into 8-week-old C57BL / 6 mice. At each time point, mice were euthanized and blood was collected via cardiac puncture. Plasma was then isolated, followed by extraction of compounds via protein precipitation and concentrations determined using LC / MS / MS. Figure 1 shows the pharmacokinetic profile of subcutaneous injection of StAMP 4 (Table A) at 20 mg / kg in mice using various StAMP:lipid ratios. "Lipid" = DSPE-MPEG (2000).

[0335] Antimicrobial activity and nephrotoxicity: Table E below shows the results of antimicrobial activity and / or nephrotoxicity assays for formulations containing various StAMP:lipid (w / w) ratios. In Table E, StAMP is StAMP 4 (Table A) and "Lipid" = DSPE-MPEG (2000). Table E. Antimicrobial Activity and / or Nephrotoxicity of StAMP-Lipid Formulations [Table E]

[0336] In Vivo Studies FIG. 2 shows the efficacy of StAMP 26 (Table A) in a P. aeruginosa pneumonia model in neutropenic mice 4 hours after treatment. Neutropenia was established via intraperitoneal administration of cyclophosphamide at doses of 150 mg / kg and 100 mg / kg on days -4 and -1, respectively. On day 0, 10 5~6 Animals were inoculated intranasally with P. aeruginosa (PA01) suspended in saline at a concentration of CFU / mL. Two hours after inoculation, a dose of StAMP 26 (formulated in phosphate buffered saline containing DSPE-PEG(2000) amine) or vehicle (phosphate buffered saline) was administered intraperitoneally, control animals were euthanized, and lungs were harvested for colony counts. Six hours after inoculation, animals were euthanized, and lungs were harvested for colony counts.

[0337] FIG. 3 shows the efficacy of StAMP 46 (Table A) in a P. aeruginosa neutropenic pneumonia mouse model. Neutropenia was established via intraperitoneal administration of cyclophosphamide at doses of 150 mg / kg and 100 mg / kg on days -4 and -1, respectively. On day 0, 10 5~6 Animals were inoculated intranasally with P. aeruginosa (PA01) suspended in saline at a concentration of CFU / mL. Two hours after inoculation, a dose of StAMP 46 (formulated in phosphate buffered saline containing DSPE-PEG(2000)amine) or vehicle (phosphate buffered saline containing DSPE-PEG(2000)amine) was administered intraperitoneally, control animals were euthanized, and lungs were harvested for colony counts. Six hours after inoculation, animals were euthanized, and lungs were harvested for colony counts.

[0338] FIG. 4 shows the overnight efficacy of StAMP 26 and StAMP 46 in a P. aeruginosa neutropenic pneumonia mouse model. Neutropenia was established via intraperitoneal administration of cyclophosphamide at doses of 150 mg / kg and 100 mg / kg on days -4 and -1, respectively. On day 0, 10 5~6 Animals were inoculated intranasally with P. aeruginosa (PA01) suspended in saline at a concentration of CFU / mL. Two hours after inoculation, a dose of StAMP 26 or StAMP 46 (formulated in phosphate buffered saline containing DSPE-PEG(2000)amine) or vehicle (phosphate buffered saline containing DSPE-PEG(2000)amine) was administered intraperitoneally, control animals were euthanized, and lungs were collected for colony counting. Six hours after inoculation, a second dose of StAMP 26 or StAMP 46 (formulated in phosphate buffered saline containing DSPE-PEG(2000)amine) or vehicle (phosphate buffered saline containing DSPE-PEG(2000)amine) was administered intraperitoneally. Eighteen hours after inoculation, animals were euthanized, and lungs were collected for colony counting.

[0339] Additional Aspects Additional aspects of the present disclosure are provided according to the following numbered paragraphs: 1. Amino acid sequence:

number

[0340] 2. Amino acid sequence:

number

[0341] 3. Amino acid sequence:

number

[0342] 4. The peptide of any one of paragraphs 1 to 3, wherein the amino acid sequence includes substitutions of 1 to 7 amino acids (inclusive), or a pharma- ceutically acceptable salt thereof.

[0343] 5. The peptide of any one of paragraphs 1 to 4, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence includes substitutions of 1 to 5 amino acids (inclusive).

[0344] 6. The peptide of any one of paragraphs 1 to 5, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence includes one or two amino acid substitutions.

[0345] 7. The peptide of any one of paragraphs 1-5, wherein the amino acid sequence comprises one to five amino acid (inclusive) substitutions, independently, at K4, H7, S8, G18, or E19, or a pharma- ceutically acceptable salt thereof.

[0346] 8. The peptide of any one of paragraphs 1-7, wherein the amino acid sequence comprises one or two amino acid substitutions, independently, at K4, H7, S8, G18, or E19, or a pharma- ceutically acceptable salt thereof.

[0347] 9. The peptide of any one of paragraphs 1-8, wherein the amino acid sequence comprises a substitution of an amino acid at H7, or a pharma- ceutically acceptable salt thereof.

[0348] 10. The peptide of any one of paragraphs 1 to 9, wherein the amino acid sequence comprises a substitution of an amino acid at S8, or a pharma- ceutically acceptable salt thereof.

[0349] 11. The peptide of any one of paragraphs 1 to 10, wherein the amino acid sequence comprises amino acid substitutions at both H7 and S8, or a pharma- ceutically acceptable salt thereof.

[0350] 12. The peptide of any one of paragraphs 1-11, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at H7 is selected from H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, and H7hArg.

[0351] 13. The peptide of paragraph 12, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at H7 is H7K.

[0352] 14. The peptide of any one of paragraphs 1-13, wherein the amino acid substitution at S8 is selected from S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, and S8hArg, or a pharma- ceutically acceptable salt thereof.

[0353] 15. The peptide of paragraph 14, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at S8 is S8K.

[0354] 16. The peptide of any one of paragraphs 1 to 15, wherein the amino acid sequence comprises a substitution of amino acid at G18, or a pharma- ceutically acceptable salt thereof.

[0355] 17. Amino acid substitutions at G18 are G18W, G18V, G18L, G18Y, G18F, G18T, and G18F. 1 , G18F 2 , G18F 4 , G18F 5 , G18F3 17. The peptide of paragraph 16, wherein the peptide is selected from: G18S, or a pharma- ceutically acceptable salt thereof.

[0356] 18. The peptide of paragraph 17, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at G18 is G18W.

[0357] 19. The peptide of any one of paragraphs 1 to 18, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence comprises an amino acid substitution at E19.

[0358] 20. The peptide of paragraph 19, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at E19 is selected from E19W, E19D, E19Q, and E19N.

[0359] 21. The peptide of any one of paragraphs 1 to 20, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence comprises an amino acid substitution at K4.

[0360] 22. The peptide of paragraph 21, or a pharma- ceutically acceptable salt thereof, wherein the amino acid substitution at K4 is selected from K4S, K4Dab, K4Orn, K4Dap, K4R, and K4hArg.

[0361] 23. The peptide of any one of paragraphs 10 to 22, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence comprises a substitution of the amino acids H7K and S8K.

[0362] 24. The peptide of any one of paragraphs 1 to 23, or a pharma- ceutically acceptable salt thereof, comprising one or more amino acid substitutions selected from G13S, A15S, G18S, and A21S.

[0363] 25. Amino acid sequence:

number

[0364] 26. Amino acid sequence:

number

[0365] 27. The peptide of paragraph 25 or 26, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence includes substitutions of 1 to 5 amino acids (inclusive).

[0366] 28. The peptide of any one of paragraphs 25 to 27, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence includes one or two amino acid substitutions.

[0367] 29. The peptide of any one of paragraphs 25 to 28, wherein the amino acid sequence includes 1 or 2 amino acids substituted by W, or a pharma- ceutically acceptable salt thereof.

[0368] 30. Amino acid sequence:

number

[0369] 31. Amino acid sequence:

number

[0370] 32. Amino acid sequence:

number

[0371] 33. Amino acid sequence:

number

[0372] 34. The peptide of any one of paragraphs 30 to 33, wherein the amino acid sequence optionally includes one or two amino acid substitutions.

[0373] 35. The peptide of any one of paragraphs 1 to 34, or a pharma- ceutically acceptable salt thereof, wherein one or more of K is independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg.

[0374] 36. The peptide of paragraph 35, or a pharma- ceutically acceptable salt thereof, wherein each K is independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg.

[0375] 37. One or more of F may be independently 1 , F 5 , F 4 , F 2 , and F 3 37. The peptide of any one of paragraphs 1 to 36, wherein the amino acid is selected from the group consisting of:

[0376] 38. Each of F is independently F 1 , F 5 , F 4 , F 2 , and F 3 38. The peptide of paragraph 37, or a pharma- ceutically acceptable salt thereof, wherein the peptide is substituted with an amino acid selected from:

[0377] 39. The peptide of any one of paragraphs 1 to 38, or a pharma- ceutically acceptable salt thereof, comprising a small molecule, a lipophilic group, or a polymer conjugated to the C-terminus of the peptide.

[0378] 40. The peptide of paragraph 39, or a pharma- ceutically acceptable salt thereof, wherein the lipophilic group is a lipid or a fatty acid.

[0379] 41. The peptide of paragraph 39, or a pharma- ceutically acceptable salt thereof, wherein the peptide comprises PEG conjugated to the C-terminus.

[0380] 42. The peptide of paragraph 41, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus comprises PEG3 or (PEG3)2.

[0381] 43. The peptide of any one of paragraphs 1 to 42, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated.

[0382] 44. The peptide of paragraph 43, wherein the C-terminus is amidated with -NH2.

[0383] 45. The peptide has at its C-terminus the formula: -NH(CH2CH2O) 1~20 44. The peptide of paragraph 43, or a pharma- ceutically acceptable salt thereof, which is amidated with a group represented by CH2CH2CONH2.

[0384] 46. ​​The peptide of paragraph 45, or a pharma- ceutically acceptable salt thereof, wherein the C-terminus is amidated with a group selected from the following: -NHCH2CH2OCH2CH2CONH2, -NH(CH2CH2O)2-CH2CH2CONH2, -NH(CH2CH2O)3-CH2CH2CONH2, -NH(CH2CH2O)4- CH2CH2CONH2, and -NH(CH2CH2O)5-CH2CH2CONH2.

[0385] 47. The peptide of any one of paragraphs 1 to 38, or a pharma- ceutically acceptable salt thereof, comprising an amino acid or peptide conjugated to the C-terminus of the peptide.

[0386] 48. The following amino acid sequence conjugated to the C-terminus of a peptide:

number

[0387] 49. The peptide of paragraph 39, or a pharma- ceutically acceptable salt thereof, comprising a polymyxin conjugated to the C-terminus of the peptide.

[0388] 50. The peptide of any one of paragraphs 1 to 49, or a pharma- ceutically acceptable salt thereof, wherein the peptide is 100 amino acids or less in length.

[0389] 51. The peptide of any one of paragraphs 1 to 50, or a pharma- ceutically acceptable salt thereof, wherein the peptide is 30 amino acids or less in length.

[0390] 52. The peptide of any one of paragraphs 1-3, comprising one of the following amino acid sequences: SEQ ID NOs: 4-39, 59-77, and 81-100; or a pharma- ceutically acceptable salt thereof.

[0391] 53. The peptide of paragraph 52, or a pharma- ceutically acceptable salt thereof, wherein the peptide is represented by one of SEQ ID NOs: 4-39, 59-77, or 81-100; and the C-terminus is amidated with -NH2.

[0392] 54. The peptide of paragraph 25 or 26, comprising one of the following amino acid sequences: SEQ ID NOs: 40-53; or a pharma- ceutically acceptable salt thereof.

[0393] 55. The peptide of paragraph 54, or a pharma- ceutically acceptable salt thereof, wherein the peptide is represented by one of SEQ ID NOs: 40 to 53; and the C-terminus is amidated with -NH2.

[0394] 56. A peptide having the following amino acid sequence:

number

[0395] 57. The peptide of paragraph 56, wherein the peptide is represented by SEQ ID NO:3; and the C-terminus is amidated with -NH2, or a pharma- ceutically acceptable salt thereof.

[0396] 58. The peptide of paragraph 32 or 33, comprising one of the following amino acid sequences: SEQ ID NOs: 102-105; or a pharma- ceutically acceptable salt thereof.

[0397] 59. The peptide of any one of paragraphs 32, 33 or 58, or a pharma- ceutically acceptable salt thereof, wherein the peptide is represented by any one of SEQ ID NOs: 101-105; and the C-terminus is amidated with -NH2.

[0398] 60. Cross-linking is done with amino acid X. 1 , X 2 , X 3 , and X 4 or a pharma- ceutically acceptable salt thereof.

[0399] 61. The peptide of any one of the preceding paragraphs, wherein each bridge is independently an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, an optionally substituted carbocyclylene, an optionally substituted heterocyclylene, an optionally substituted arylene, an optionally substituted heteroarylene, an optionally substituted acylene, or any combination thereof, or a pharma- ceutically acceptable salt thereof.

[0400] 62. The peptide of paragraph 61, or a pharma- ceutically acceptable salt thereof, wherein each bridge is independently a hydrocarbon bridge selected from optionally substituted alkylene, optionally substituted alkenylene, and optionally substituted alkynylene.

[0401] 63. Each bridge independently has the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive, or a pharma- ceutically acceptable salt thereof.

[0402] 64. The peptide of paragraph 63, or a pharma- ceutically acceptable salt thereof, wherein the sum of two n's on the same bridge is 6.

[0403] 65.X 1 , X 2 , X 3 , and X 4 is independently an α,α-disubstituted amino acid, or a pharma- ceutically acceptable salt thereof.

[0404] 66.X 1 and X 2 and X 3 and X 4 are each independently connected by a bridge to have the formula: [ka] where α represents the α-carbon of the amino acid; and where R 1 each independently represents an optionally substituted C 1~6 The peptide of any one of the preceding paragraphs, or a pharma- ceutically acceptable salt thereof, wherein:

[0405] 67.R 1 67. The peptide of paragraph 66, or a pharma- ceutically acceptable salt thereof, wherein at least one of is methyl.

[0406] 68.X 1 and X 2 and X 3 and X 4 are connected to each other by a bridge and have the following formula: [ka] wherein α represents the α-carbon of an amino acid; or a pharma- ceutically acceptable salt thereof.

[0407] 69. The peptide of any one of the preceding paragraphs, or a pharma- ceutically acceptable salt thereof, wherein each bridge is independently from about 10 Å to about 16 Å in length, inclusive.

[0408] 70. The peptide of any one of the preceding paragraphs, or a pharma- ceutically acceptable salt thereof, wherein the length of each bridge is approximately equal to the length of 5 to 13 carbon-carbon bonds (inclusive).

[0409] 71. The peptide of any one of the preceding paragraphs, wherein at least one bridge spans at least one turn of the α-helix of the peptide, or a pharma- ceutically acceptable salt thereof.

[0410] 72. The peptide of paragraph 71, or a pharma- ceutically acceptable salt thereof, wherein at least one crosslink stabilizes an α-helix of the peptide.

[0411] 73. The peptide of paragraph 71 or 72, or a pharma- ceutically acceptable salt thereof, wherein the peptide has increased alpha-helicity compared to the corresponding uncrosslinked peptide.

[0412] 74.X 1 , X 2 , X 3 , and X 4 independently, the peptide of any one of the preceding paragraphs, or a pharma- ceutically acceptable salt thereof, which comprises an α-side chain that comprises a reactive moiety.

[0413] 75. The peptide of any one of the preceding paragraphs, or a pharma- ceutically acceptable salt thereof, wherein the reactive moieties are independently selected from alkenes and alkynes.

[0414] 76.X 1 , X 2 , X 3 , and X 4 are independently of the formula: [ka] wherein each n is independently an integer from 1 to 10, inclusive; or a pharma- ceutically acceptable salt thereof.

[0415] 77.X 1 , X 2 , X 3 , and X 4 is independently an α,α-disubstituted amino acid, or a pharma- ceutically acceptable salt thereof.

[0416] 78.X 1 , X 2 , X 3 , and X 4 are each independently represented by the formula: [ka] where α represents the α-carbon of an amino acid; and R 1 Each of the groups may be an optionally substituted C 1~6 80. The peptide of paragraph 76 or 77, or a pharma- ceutically acceptable salt thereof, comprising:

[0417] 79.R 1 80. The peptide of paragraph 78, or a pharma- ceutically acceptable salt thereof, wherein at least one of is methyl.

[0418] 80.X 1 , X 2 , X 3 , and X 4 Each of the formula: [ka] or a pharma- ceutically acceptable salt thereof.

[0419] 81. A pharmaceutical composition comprising the peptide of any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

[0420] 82. A method of treating an infectious disease in a subject, the method comprising administering to the subject a peptide of any one of paragraphs 1-73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0421] 83. The method of paragraph 82, wherein the infectious disease is a bacterial infection, a viral infection, a protozoal infection, or a fungal infection.

[0422] 84. A method of treating a bacterial infection in a subject, the method comprising administering to the subject a peptide of any one of paragraphs 1-73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0423] 85. The method of paragraph 84, wherein the bacterial infection is a gram-negative bacterial infection.

[0424] 86. The method of paragraph 84 or 85, wherein the bacterial infection is an antibiotic-resistant bacterial infection.

[0425] 87. The method of any one of paragraphs 84-86, wherein the bacterial infection is a gram-negative, antibiotic-resistant bacterial infection.

[0426] 88. The method of paragraph 86 or 87, wherein the antibiotic-resistant bacterial infection is caused by bacteria that are resistant to one or more antibiotics selected from the group consisting of polymyxins, carbapenems, aminoglycosides, vancomycin, methicillin, clarithromycin, cephalosporins, penicillins, ampicillin, fluoroquinolones, tetracyclines, ceftazidime, ceftriaxone, ciprofloxacin, gentamicin, tobramycin, and beta-lactams.

[0427] 89. The method of any one of paragraphs 84-88, wherein the bacterial infection is caused by E. coli, A. baumannii, P. aeruginosa, or K. pneumoniae.

[0428] 90. The method of any one of paragraphs 82 to 89, wherein the peptide has reduced nephrotoxicity compared to a reference.

[0429] 91. The method of paragraph 90, wherein the peptide has reduced nephrotoxicity compared to a corresponding peptide comprising SEQ ID NO: 1 or 2, or compared to pexiganan.

[0430] 92. The method of any one of paragraphs 82 to 91, wherein the peptide has reduced hepatotoxicity compared to a reference.

[0431] 93. The method of paragraph 92, wherein the peptide has reduced hepatotoxicity compared to a corresponding peptide comprising SEQ ID NO: 1 or 2, or compared to pexiganan.

[0432] 94. The method of any one of paragraphs 82 to 93, wherein the peptide has reduced hemolytic activity compared to a reference.

[0433] 95. The method of paragraph 94, wherein the peptide has reduced hemolytic activity compared to a corresponding peptide comprising SEQ ID NO: 1 or 2, or compared to melittin.

[0434] 96. The method of any one of paragraphs 82-95, wherein the peptide, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof is administered intravenously.

[0435] 97. The method of any one of paragraphs 82 to 96, wherein the subject is a human.

[0436] 98. A method for killing and / or inhibiting the growth of bacteria, the method comprising contacting the bacteria with a peptide of any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0437] 99. The method of paragraph 98, wherein the bacterium is E. coli, A. baumannii, P. aeruginosa, or K. pneumoniae.

[0438] 100. A method for selectively killing and / or inhibiting the growth of microbial cells over mammalian cells, the method comprising contacting microbial cells and mammalian cells with a peptide of any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0439] 101. A method for selectively lysing microbial cells over mammalian cells, the method comprising contacting the microbial cells and the mammalian cells with any one of the peptides of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0440] 102. The method of paragraph 100 or 101, wherein the microbial cells are bacterial cells.

[0441] 103. The method of paragraph 102, wherein the bacterial cell is an E. coli cell, an A. baumannii cell, a P. aeruginosa cell, or a K. pneumoniae cell.

[0442] 104. A peptide of any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in a method of any one of the preceding paragraphs.

[0443] 105. Use of a peptide according to any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the preparation of a medicament.

[0444] 106. A kit comprising a peptide of any one of paragraphs 1 to 73, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof; and, optionally, instructions for use.

[0445] 107. A method for preparing a cross-linked peptide of any one of the preceding paragraphs, comprising: 1 and X 2 and X 3 and X 4 The method includes reacting the compound under conditions sufficient to form a crosslink connecting the compound and the compound.

[0446] 108. The method of paragraph 107, wherein the reacting step involves a ring-closing metathesis (RCM) reaction.

[0447] 109. A formulation comprising a stapled antimicrobial peptide (StAMP) and one or more lipids.

[0448] 110. The formulation of paragraph 109, wherein the formulation is a micellar formulation, a liposomal formulation, or a lipid nanoparticle formulation.

[0449] 111. The formulation of paragraph 109 or 110, wherein StAMP is a peptide of any one of the preceding paragraphs.

[0450] 112. The formulation of any one of paragraphs 109 to 111, wherein at least one of the lipids in paragraph 1 or more comprises a phospholipid.

[0451] 113. The formulation of paragraph 112, wherein at least one of the lipids 1 or more is a pegylated phospholipid.

[0452] 114. The formulation of paragraph 113, wherein the PEGylated phospholipid is DSPE-PEG.

[0453] 115. The formulation of paragraph 114, wherein the PEGylated phospholipid is DSPE-MPEG amine or DSPE-PEG amine.

[0454] 116. The formulation of paragraph 115, wherein the pegylated phospholipid is DSPE-MPEG(2000)amine or DSPE-PEG(2000)amine.

[0455] 117. The formulation of any one of paragraphs 109-116, comprising a peptide:lipid ratio of about 1:1 to about 1:25 (w / w), inclusive.

[0456] 118. The formulation of any one of paragraphs 109-117, comprising a peptide:lipid ratio of about 1:2.5 to about 1:20 (w / w), inclusive.

[0457] 119. The formulation of any one of paragraphs 109-118, comprising a peptide:lipid ratio of about 1:2.5, about 1:5, about 1:10, or about 1:20 (w / w).

[0458] Equivalents and Scope In the claims, unless indicated to the contrary or otherwise clear from the context, articles such as "a," "an," and "the" can mean one or more than one. Unless indicated to the contrary or otherwise clear from the context, a claim or description including "or" between one or more members of a group is considered to be satisfied if one, more than one, or all of the members of the group are present in, used in, or otherwise involved in a given product or process. The present disclosure includes embodiments in which exactly one member of a group is present in, used in, or otherwise involved in a given product or process. The present disclosure includes embodiments in which more than one or all of the members of a group are present in, used in, or otherwise involved in a given product or process.

[0459] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the enumerated claims are introduced into another claim. For example, any claim that is dependent on another claim may be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented in the Markush group format as an example list, each subgroup of elements is also disclosed, and any element(s) may be removed from the group. In general, where the disclosure or aspects of the disclosure are said to include certain elements and / or features, it should be understood that certain aspects of the disclosure or aspects of the disclosure consist of or consist essentially of such elements and / or features. For purposes of simplicity, those aspects have not been specifically submitted as such in the application. It is also noted that the terms "comprise" and "contain" are intended to be open-ended, permitting the inclusion of additional elements or steps. Where ranges are given, the endpoints are included. Furthermore, unless otherwise indicated or otherwise apparent from the context and the understanding of one of ordinary skill in the art, values ​​expressed as ranges can, in different aspects of this disclosure, take any particular value or subrange within the claimed range to one tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0460] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. In the event of a conflict between any of the incorporated references and this specification, this specification shall control. In addition, any specific aspects of this disclosure that are in the prior art may be expressly excluded from any one or more of the claims. Such aspects may be excluded even if the exclusion is not expressly stated in this application, since they are deemed to be known to those of ordinary skill in the art. Any specific aspects of this disclosure may be excluded from any claim for any reason, whether or not related to the existence of prior art.

[0461] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. In the event of a conflict between any of the incorporated references and this specification, this specification shall control. In addition, any specific aspects of this disclosure that are in the prior art may be expressly excluded from any one or more of the claims. Such aspects may be excluded even if the exclusion is not expressly stated in this application, since they are deemed to be known to those of ordinary skill in the art. Any specific aspects of this disclosure may be excluded from any claim for any reason, whether or not related to the existence of prior art.

[0462] Those skilled in the art will recognize, or be able to ascertain, many equivalents to the specific embodiments described herein, at most using routine experimentation. The scope of the embodiments described herein is not intended to be limited to the above specification, but rather is set forth in the appended claims. Those skilled in the art will appreciate that various changes and modifications of this description can be made without departing from the spirit or scope of the present disclosure, as defined in the following claims.

Claims

1. Amino acid sequence: [0010] or a pharma- ceutically acceptable salt thereof, wherein: X 1 , X 2 , X 3 , and X 4 is, independently, an amino acid; X 1 and X 2 and are connected via bridges, and X 3 and X 4 and are connected via a bridge; and The amino acid sequence is expressed as X, which is a substitution of 1 to 9 amino acids (inclusive). 1 , X 2 , X 3 , and X 4 Inclusion at a position other than; with the proviso that at least one amino acid substitution is at H7, S8, G18, or E19; and with the proviso that the amino acid substitution at G18 is not G18H. The peptide, or a pharma- ceutically acceptable salt thereof.

2. Amino acid sequence: [0025] or a pharma- ceutically acceptable salt thereof, wherein: X 1 , X 2 , X 3 , and X 4 is, independently, an amino acid; X 1 and X 2 each independently contains a reactive moiety capable of forming a crosslink with the others, and X 3 and X 4 each independently contains a reactive moiety capable of forming a crosslink with the other; and The amino acid sequence is expressed as X, which is a substitution of 1 to 9 amino acids (inclusive). 1 , X 2 , X 3 , and X 4 Inclusion at a position other than; with the proviso that at least one amino acid substitution is at H7, S8, G18, or E19; and with the proviso that the amino acid substitution at G18 is not G18H. The peptide, or a pharma- ceutically acceptable salt thereof.

3. Amino acid sequence: [0030] 3. The peptide of claim 1 or 2, or a pharma- ceutically acceptable salt thereof, comprising: wherein the amino acid sequence includes substitutions of 1 to 11 amino acids (inclusive), or a pharma- ceutically acceptable salt thereof.

4. 4. The peptide of any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, wherein the amino acid sequence comprises amino acid substitutions at H7, S8, G18, K4, and / or E19.

5. the amino acid substitutions in H7 are selected from H7K, H7W, H7Dab, H7Orn, H7Dap, H7R, and H7hArg; The amino acid substitution at S8 is selected from S8K, S8W, S8N, S8Q, S8T, S8Y, S8Dab, S8Orn, S8Dap, S8R, and S8hArg; The amino acid substitutions at G18 are G18W, G18V, G18L, G18Y, G18F, G18T, and G18F. 1 , G18F 2 , G18F 4 , G18F 5 , G18F 3 and G18S; the amino acid substitution at K4 is selected from K4S, K4Dab, K4Orn, K4Dap, K4R, and K4hArg; and the amino acid substitution at E19 is selected from E19W, E19D, E19Q, and E19N; 5. The peptide according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof.

6. 6. The peptide of claim 1, or a pharma- ceutically acceptable salt thereof, wherein one or more of K is independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg.

7. 7. The peptide of claim 6, or a pharma- ceutically acceptable salt thereof, wherein each K is independently replaced by an amino acid selected from Orn, Dab, Dap, R, and hArg.

8. The peptide has the following amino acid sequence: [0045] [0050] 4. The peptide according to claim 1, or a pharma- ceutically acceptable salt thereof, comprising one of:

9. The peptide is represented by one of SEQ ID NOs: 4-39, 59-77, or 81-100; and wherein the C-terminus is -NH 2 9. The peptide of claim 8, or a pharma- ceutically acceptable salt thereof, which is amidated with:

10. The crosslink is the amino acid X 1 , X 2 , X 3 , and X 4 10. The peptide of any one of claims 1 and 3 to 9, or a pharma- ceutically acceptable salt thereof, wherein the peptide is attached to the alpha position of

11. Each bridge independently has the formula: 【Chemistry 1】 wherein each n is independently an integer from 1 to 10, inclusive, or a pharma- ceutically acceptable salt thereof.

12. X 1 and X 2 and X 3 and X 4 and each independently are connected by a bridge to form a compound of the formula: 【Chemistry 2】 where α represents the α-carbon of the amino acid; and where R 1 each independently represents an optionally substituted C 1～6 is alkyl, Preferably, X 1 and X 2 and X 3 and X 4 and each are connected by a bridge to form a compound of the formula: 【Chemistry 3】 where α represents the α-carbon of the amino acid.

12. A peptide according to any one of claims 1 and 3 to 11, or a pharma- ceutically acceptable salt thereof.

13. A pharmaceutical composition comprising the peptide according to any one of claims 1 to 12, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

14. (i) a method of treating an infectious disease in a subject; (ii) a method of treating a bacterial infection in a subject; (iii) a method for killing and / or inhibiting the growth of bacteria in a subject; (iv) a method for selectively killing and / or inhibiting the growth of microbial cells over mammalian cells in a subject; and / or (v) A method for selectively lysing microbial cells over mammalian cells in a subject. A peptide according to any one of claims 1 to 12, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in

15. A method for preparing a crosslinked peptide according to any one of claims 1 and 3 to 12, comprising: 1 and X 2 and X 3 and X 4 reacting under conditions sufficient to form a bridge connecting

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