Antiviral structured pegylated influenza peptides and uses thereof

Abstract

The present disclosure provides structured stapled peptides useful for treating and / or preventing influenza infection, as well as methods of treating and / or preventing influenza infection.

Description

Related applications

[0001] This application claims priority and benefit to U.S. Application No. 63 / 599,361, filed November 15, 2023, the entire contents of which are hereby incorporated by reference. Background Technology

[0002] Influenza infection is one of the most common viral infections worldwide each year. Influenza virus infects host cells when its lipid envelope fuses with the host cell's endosomal membrane to release the viral genome into the cell. Specifically, the envelope glycoprotein hemagglutinin (HA) mediates the binding of the viral envelope to the host cell's endosomal membrane, leading to membrane fusion, viral genome replication, and infection. The influenza HA is divided into two domains, HA1 and HA2, where HA1 mediates host cell receptor binding and HA2 mediates the fusion of the viral membrane with the endosomal membrane. The fusion of the influenza virus particle with the host cell's endosomal membrane is mediated by a conformational change in HA, which is driven by a lower pH in the endosomal region.

[0003] Cryo EM studies at pH 5.5 (endosome pH) highlighted several conformational changes in influenza HA during host cell fusion. In the final conformational change, the C-terminal helix of the HA2 domain (residues 106 to 125) folds back into the HR2 portion of the longer HA2 helix, bringing the viral membrane close to the endosomal membrane and fusion occurring. Furthermore, the C-terminal helix of HA2 is highly conserved across influenza A subtypes.

[0004] Despite the availability of flu vaccines and treatments, there are still millions of cases of flu illness, hundreds of thousands of hospitalizations due to the flu, and thousands of deaths from the flu each year. Summary of the Invention

[0005] This disclosure provides modified peptides that can be used to treat and / or prevent influenza virus infections (e.g., influenza A or influenza B infections). In some embodiments, the provided peptides are cross-linked (e.g., stapled) to stabilize the helical structure of the peptide. The peptides provided herein are particularly advantageous because they are not limited to treating or preventing a specific influenza subtype and can therefore be used to treat or prevent more than one influenza subtype.

[0006] This disclosure covers the understanding that blocking the final conformational change of HA can inhibit the fusion of influenza with host cells and provides a novel strategy for the prevention and / or treatment of influenza infection not limited to specific influenza subtypes. The modified peptides provided herein are believed to prevent this conformational change in the HA2 domain of influenza HA (e.g., the C-terminus of HA2 helically folds back into the HR2 region), and thus prevent the fusion of influenza with host cells.

[0007] In some embodiments, this disclosure provides a modified peptide of Formula I:

[0008] I, Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 Xaa, x, w, and y are as defined in this paper.

[0009] This disclosure also provides methods for treating and / or preventing influenza virus infection, the method comprising administering to a patient in need a therapeutically effective amount of the modified peptides provided herein, such as peptides of formula I, or pharmaceutically acceptable salts thereof. Attached Figure Description

[0010] Figure 1 This is a schematic diagram illustrating the mapping of C-terminal helical interactions on the sequences of influenza A subtypes. Common sequences for each subtype are shown and colored to reflect conservation. Most residues of a given subtype are conserved by 95% or higher. Proposed therapeutic peptides are shown in boxes, and residues interacting between the C-terminal helix and HA2 are in bold.

[0011] Figure 2 This is a schematic diagram illustrating the interaction mapping of the C-terminal helix of H3N3. The sequence of the loop and helical regions of the C-terminal helix is ​​shown. Connections are made within or to the outer boxes of the C-terminal helix. Connections involving hydrogen bonds, hydrophobic interactions, and potential pinning sites are shown. Residues within the boxes illustrate the conservation of C-terminal helical contacts for H1N1, H3N2, and influenza B.

[0012] Figure 3 This illustrates a synthetic scheme for resin-derivating a stapled peptide sequence with a PEG-linked cholesterol moiety. Detailed Implementation

[0013] Compounds and definitions The compounds disclosed herein (e.g., modified peptides) include those generally described herein and are further illustrated by the classes, subclasses and species disclosed herein. Unless otherwise indicated, the following definitions shall apply as used herein.

[0014] For the purposes of this disclosure, chemical elements are identified according to the periodic table, CAS version, Handbook of Chemistry and Physics, 75th edition. Furthermore, the general principles of organic chemistry are described in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999; and "March's Advanced Organic Chemistry," 5th edition, edited by Smith, MB and March, J., John Wiley & Sons, New York: 2001.

[0015] Unless otherwise stated, the structures described herein are intended to include all stereoisomers (e.g., enantiomers or diastereomers) of the structure, as well as all geometric or conformational isomers of the structure. For example, the R and S configurations of each stereocenter are considered part of this disclosure. Therefore, single stereochemical isomers of the provided compounds, as well as mixtures of enantiomers, diastereomers, and geometric isomers (or conformations), are all within the scope of this disclosure. Unless otherwise stated, all tautomers of the provided compounds are within the scope of this disclosure.

[0016] Unless otherwise indicated, the structures described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having this structure (including those with hydrogen replaced by deuterium or tritium, or those with...) 13 C or 14 Compounds enriched with carbon (carbon substitutes for carbon) are within the scope of this disclosure.

[0017] As used herein, the term "aliphatic" or "aliphatic group" means a straight-chain (i.e., unbranched) or branched substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon (also referred to herein as "cycloaliphatic") that is fully saturated or contains one or more unsaturated units but is not aromatic, having a single connection point to the rest of the molecule. Unless otherwise specified, an aliphatic group contains 1 to 6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1 to 4 aliphatic carbon atoms. In still other embodiments, an aliphatic group contains 1 to 3 aliphatic carbon atoms, and in yet another embodiment, an aliphatic group contains 1 to 2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" refers to a fully saturated or non-aromatic monocyclic C3-C6 hydrocarbon that contains one or more unsaturated units. Suitable aliphatic groups include, but are not limited to, straight-chain or branched substituted or unsubstituted alkyl, alkenyl, ynyl and their hybrids, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0018] The term "alkyl" as used alone or as part of a larger part refers to a saturated, optionally substituted, straight-chain or branched hydrocarbon group having (unless otherwise specified) 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 3, or 1 to 2 carbon atoms (e.g., C12, C23, C14, C23, C2 ... 1-12 C 1-10 C 1-8 C 1-6 C 1-4 C 1-3 Or C 1-2 Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.

[0019] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., –(CH2). n – where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. The substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those described herein with respect to substituted aliphatic groups.

[0020] The term "alkenyl" used alone or as part of a larger part refers to an optionally substituted straight-chain or branched hydrocarbon chain having at least one double bond and having (unless otherwise specified) 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C4, C5, C6, C7, C8, C9 ... 2-12 C 2-10 C 2-8 C 2-6 C2-4 Or C 2-3 Exemplary alkenyl groups include vinyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl.

[0021] The term "alkenyl" refers to a divalent alkenyl group. A substituted alkenyl chain is a divalent hydrocarbon chain containing at least one double bond, wherein one or more hydrogen atoms are replaced by substituents. Suitable substituents include those described herein with respect to substituted aliphatic groups.

[0022] The term "alkynyl" used alone or as part of a larger part refers to an optionally substituted straight-chain or branched hydrocarbon group having at least one triple bond and having (unless otherwise specified) 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C4, C5, C6, C7, C8, C9 ... 2-12 C 2-10 C 2-8 C 2-6 C 2-4 Or C 2-3 Exemplary alkynyl groups include ethynyl, propynyl, butynyl, penynyl, hexynyl, and heptynyl.

[0023] As used herein, the term "ynynyl" refers to a divalent ynyl group. A substituted ynynyl chain is a divalent hydrocarbon chain containing at least one triple bond, wherein one or more hydrogen atoms are replaced by substituents. Suitable substituents include those described herein with respect to substituted aliphatic groups.

[0024] The term "aryl," used alone or as part of a larger part such as in "aralkyl," "aralkyloxy," or "aryloxyalkyl," refers to a monocyclic or bicyclic system having a total of six to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In some embodiments of this disclosure, "aryl" refers to aromatic ring systems including, but not limited to, phenyl, naphthyl, anthracene, etc., which may carry one or more substituents. Unless otherwise specified, the "aryl" group is a hydrocarbon.

[0025] As used herein, the term "bicyclic" or "bicyclic system" refers to any bicyclic system, i.e., a carbon ring or heterocyclic ring, saturated or having one or more unsaturated units, with one or more common atoms between the two rings of the system. Therefore, the term includes any permitted ring fusion, such as AdjacentFusion or spirocyclic. As used herein, the term "heterobicyclic" is a subset of "bicyclic," requiring one or more heteroatoms to be present in one or both rings of the bicyclic ring. Such heteroatoms may be present at ring junctions and optionally substituted, and may be selected from nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates), boron, etc. In some embodiments, the bicyclic group has 7 to 12 ring members and 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0026] The term "halogen" or "halogenated" refers to F, Cl, Br, or I.

[0027] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; any basic nitrogen; any quaternized form of basic nitrogen; or a substituted nitrogen of a heterocycle, such as N (e.g., in 3,4-dihydro-2-) H -pyrrole group), NH (as in pyrroleyl group) or NR + (e.g., in N-substituted pyrroleyl groups).

[0028] The terms "heteroaryl" and "heteroaryl-" used alone or as part of a larger portion, such as "heteroarylalkyl" or "heteroarylalkoxy," refer to a group having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; a group sharing 6, 10, or 14 π electrons in a cyclic array; and having one to five heteroatoms (e.g., 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur) in addition to a carbon atom. Heteroaryl groups include, but are not limited to, thiopheneyl, furanyl, pyrroleyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, indoleazinyl, purine, naphridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroaryl-" also include groups in which the heteroaryl ring is fused with one or more aryl rings, cycloaliphatic rings, or heterocyclic rings. Non-limiting examples include indole, isoindole, benzothiamyl, benzofuranyl, dibenzofuranyl, indazole, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, terolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4 H- Quinazinyl, carbazoyl, acridineyl, phenazinyl, phenothiazinyl, phenotoxazinyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl. The heteroaryl group can be monocyclic or bicyclic. The term "heteroaryl" is used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which contains an optionally substituted ring.

[0029] As used herein, the terms “heterocycle,” “heterocyclic group,” and “heterocyclic ring” are used interchangeably and refer to a stable 3- to 7-membered monocyclic or 5- to 10-membered bicyclic heterocyclic portion that is saturated or partially unsaturated and has one or more, preferably one to four, heteroatoms as defined above, in addition to a carbon atom. When referring to the ring atom of the heterocycle, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen can be N (e.g., 3,4-dihydro-2-oxohydrogen ions). H- In pyrroleyl), NH (such as in pyrrolealkyl) or + NR (e.g.) N- (In substituted pyrrolidinyl groups). The heterocycle can be attached to its side group at any heteroatom or carbon atom to produce a stable structure, and any ring atom can optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazine, dioxacyclohexyl, dioxopentyl, diazapyryl, oxazopentyl, thiaazopentyl, morpholinyl, and thiomorpholinyl. The heterocyclic group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. Bicyclic heterocycles also include groups in which the heterocycle is fused with one or more aryl, heteroaryl, or cycloaliphatic rings. Exemplary bicyclic heterocyclic groups include indololinyl, isoidololinyl, benzo-m-dioxacyclopentenyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. Bicyclic heterocycles can also be spirocyclic systems (e.g., 7- to 11-membered spirocyclic heterocycles having one or more heteroatoms as defined above (e.g., one, two, three, or four heteroatoms) in addition to a carbon atom).

[0030] As used herein, when referring to ring moieties, the term "partially unsaturated" means a ring moieties comprising at least one double or triple bond. The term "partially unsaturated" is intended to cover rings having multiple unsaturated sites, but not to include aryl or heteroaryl moieties as defined herein.

[0031] As used in this article, the term "unsaturated" means that a part has one or more unsaturated units.

[0032] As described herein, the compounds of this disclosure may contain an "optionally substituted" moiety. Generally, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are substituted by a suitable substituent. Unless otherwise indicated, the "optionally substituted" group may have a suitable substituent at each substituted position of the group, and the substituents at each position may be the same or different when more than one position in any given structure is substituted by a substituent selected from the specified group. The combinations of substituents contemplated in this disclosure are combinations that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" means that the compound remains substantially unchanged when subjected to conditions that allow it to be generated, detected, and, in some embodiments, recovered, purified, and used for one or more purposes disclosed herein.

[0033] The suitable monovalent substituent on the substituted carbon atom of the "optionally substituted" group is independently a halogen; –(CH2) 0–4 R°;–(CH2) 0–4 OR°;-O(CH2) 0-4 R o –O–(CH2) 0–4 C(O)OR°;–(CH2) 0–4 CH(OR°)2;–(CH2) 0– 4SR°; –(CH2) 0–4 Ph, which can be substituted by R°; –(CH2) 0–4 O(CH2) 0–1 Ph, which can be substituted by R°; –CH=CHPh, which can be substituted by R°; –(CH2) 0–4 O(CH2) 0–1 -pyridyl group, which can be substituted by R°; –NO2; –CN; –N3; -(CH2) 0–4 N(R°)2;–(CH2) 0–4 N(R°)C(O)R°; –N(R°)C(S)R°; –(CH2) 0–4 N(R°)C(O)NR°2; -N(R°)C(S)NR°2; –(CH2) 0–4 N(R°)C(O)OR°; –N(R°)N(R°)C(O)R°; –N(R°)N(R°)C(O)NR°2; –N(R°)N(R°)C(O)OR°; –(CH2) 0–4 C(O)R°; –C(S)R°; –(CH2) 0–4 C(O)OR°;–(CH2) 0–4 C(O)SR°;-(CH2) 0–4 C(O)OSiR°3;–(CH2)0–4 OC(O)R°;–OC(O)(CH2) 0–4 SR°;–SC(S)SR°;–(CH2) 0–4 SC(O)R°;–(CH2) 0–4 C(O)NR°2; –C(S)NR°2; –C(S)SR°; –(CH2) 0–4 OC(O)NR°2; -C(O)N(OR°)R°; –C(O)C(O)R°; –C(O)CH2C(O)R°; –C(NOR°)R°; –(CH2) 0– 4SSR°;–(CH2) 0–4 S(O)2R°;–(CH2) 0–4 S(O)₂OR°;–(CH₂) 0–4 OS(O)2R°; –S(O)2NR°2; -(CH2) 0–4 S(O)R°; -N(R°)S(O)2NR°2; –N(R°)S(O)2R°; –N(OR°)R°; –C(NH)NR°2; –P(O)(OR°)R°; –P(O)R°2; –OP(O)R°2; –OP(O)(OR°)2; –SiR°3; –(C 1–4 (linear or branched alkylene)O–N(R°)2; or –(C 1–4 (straight-chain or branched alkylene)C(O)O–N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C 1–6 Aliphatic, –CH2Ph, –O(CH2) 0–1 Ph, -CH2- (5- to 6-membered heteroaryl ring) or a 3- to 6-membered saturated, partially unsaturated or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or, in addition to the above definitions, two independently occurring R° together with one or more intermediate atoms to form a 3- to 12-membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0034] Suitable monovalent substituents on R° (or rings formed by two independently occurring R° atoms together with their intermediate atoms) are independently halogens, –(CH2). 0–2 R ● –(halogenated R) ● ), –(CH2) 0–2 OH, –(CH2) 0–2 OR ● –(CH2) 0–2 CH(OR ● )2;-O(halogenated R ●–CN, –N3, –(CH2) 0–2 C(O)R ● –(CH2) 0–2 C(O)OH, –(CH2) 0–2 C(O)OR ● –(CH2) 0–2 SR ● –(CH2) 0–2 SH, –(CH2) 0–2 NH2、–(CH2) 0–2 NHR ● –(CH2) 0–2 NR ● 2. –NO2, –SiR ● 3. –OSiR ● 3. -C(O)SR ● 、 –(C 1–4 (straight-chain or branched alkylene)C(O)OR ● Or –SSR ● , where each R ● It is either unsubstituted or, in the case of the prefix "halogenated", substituted with only one or more halogens, and independently selected from C. 1–4 Aliphatic, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of R° include =O and =S.

[0035] Suitable divalent substituents on the saturated carbon atom of the "optionally substituted" group include the following: =O, =S, =NNR. * 2、=NNHC(O)R * =NNHC(O)OR * =NNHS(O)2R * =NR * =NOR * 、 –O(C(R) * 2)) 2–3 O – or –S(C(R) * 2)) 2– 3S–, where each independently occurring R * Selected from hydrogen, C can be substituted as defined below. 1–6 Aliphatic, or having 0 to 4 unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl rings independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents bonded to the ortho-substituted carbon of the "optionally substituted" group include: –O(CR * 2)2–3 O–, where each independently occurring R * Selected from hydrogen, C can be substituted as defined below. 1–6 Aliphatic, or having 0 to 4 unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl rings independently selected from nitrogen, oxygen, or sulfur.

[0036] R * Suitable substituents on aliphatic groups include halogens, -R ● -(halogenated R) ● -OH, -OR ● –O (halogenated R) ● ), –CN, –C(O)OH, –C(O)OR ● –NH2, –NHR ● –NR ● 2 or –NO2, where each R ● It is either unsubstituted or, in the case of the prefix "halogenated", substituted with only one or more halogens, and independently C. 1–4 Aliphatic, –CH2Ph, –O(CH2) 0– 1Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0037] Suitable substituents on the substituted nitrogen of the "optionally substituted" group include –R † –NR † 2. –C(O)R † –C(O)OR † –C(O)C(O)R † –C(O)CH2C(O)R † -S(O)2R † -S(O)2NR † 2. –C(S)NR † 2. –C(NH)NR † 2 or –N(R) † )S(O)2R † ; where each R † Independently, hydrogen can be substituted C as defined below. 1-6 Aliphatic, unsubstituted -OPh, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen, or sulfur; or, in addition to the above definitions, two independently occurring R... † Together with its intermediate atom, it forms an unsubstituted 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring with 0 to 4 independent heteroatoms selected from nitrogen, oxygen, or sulfur.

[0038] R † Suitable substituents on the aliphatic group can be halogens, -R ● -(halogenated R) ● –OH, –OR ● –O (halogenated R) ● ), –CN, –C(O)OH, –C(O)OR ● –NH2, –NHR ● –NR ● 2 or -NO2, where each R ● It is either unsubstituted or, in the case of the prefix "halogenated", substituted with only one or more halogens, and independently C. 1–4 Aliphatic, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0039] As used herein, the term "pharmaceutically acceptable salt" refers to salts that, within reasonable medical judgment, are suitable for contact with tissues of humans or other animals without undue toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berger... et al. Pharmaceutically acceptable salts are described in detail in J. Pharmaceutical Sciences, 1977, 66, 1–19.

[0040] As used herein, the term "pharmaceutically acceptable carrier" refers to any standard drug carrier, such as phosphate-buffered saline solutions, water, emulsions (…). For example Examples of carriers, such as oil / water or water / oil emulsions, and various types of wetting agents, which are suitable for contact with human and / or animal tissues within reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio. Examples of carriers, as well as stabilizers and adjuvants, are also included. See, for example , Martin, Remington's Pharmaceutical Sciences, 15th ed., Mack Publ. Co., Easton, PA

[1975] .

[0041] As used herein, the term “prevent” or “prevention”, when used in conjunction with the presence of a disease, condition, and / or symptom, means reducing the risk of developing a disease, condition, and / or symptom and / or delaying the onset of one or more features or symptoms of a disease, condition, or symptom.

[0042] As used herein, the terms "subject" and "patient" are used interchangeably and refer to an organism to be treated by the methods of the present invention. Such organisms preferably include, but are not limited to, mammals ( For example Animals, including rats, monkeys, horses, cattle, pigs, dogs, cats, etc., and preferably humans.

[0043] As used herein, the phrase “therapeutic effective amount” means the amount of active agent that effectively produces some of the desired therapeutic effect in at least one cell subpopulation of a subpopulation of animals and / or subjects, with a reasonable benefit / risk ratio applicable to any medical treatment.

[0044] As used herein, the term “treatment” (also “treatment” or “treating”) means any application of a therapy that partially or completely relieves, improves, reduces, or inhibits one or more symptoms, features, and / or causes of a particular disease, condition, and / or symptom, delays its onset, reduces its severity, and / or reduces its incidence.

[0045] Influenza peptide This disclosure provides modified peptides that can be used to treat and / or prevent influenza virus infections (e.g., influenza A or influenza B infections). In some embodiments, the provided peptides are cross-linked (e.g., stapled) to stabilize the helical structure of the peptide. The modified peptides provided herein are particularly advantageous because they are not limited to treating or preventing a specific influenza subtype and can therefore be used to treat or prevent more than one influenza subtype.

[0046] In some embodiments, this disclosure provides modified peptides comprising an amino acid sequence of an influenza HA protein or a variant thereof. In some embodiments, this disclosure provides modified peptides comprising an amino acid sequence of an influenza HA protein or a variant thereof, wherein at least two amino acids comprise side chains linked together (e.g., to form a staple).

[0047] In some embodiments, the provided modified peptide comprises an amino acid sequence substantially identical to the amino acid sequences selected from Table 1. In some embodiments, the provided modified peptide comprises an amino acid sequence substantially identical to the amino acid sequences selected from Table 1, wherein at least two amino acids of the sequence comprise side chains linked together (e.g., to form a staple).

[0048] Table 1.

[0049] In some embodiments, the provided modified peptide comprises at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98% of the amino acid sequence selected from Table 1, wherein at least two amino acids of the sequence comprise side chains linked together (e.g., to form a staple). In some embodiments, the provided modified peptide comprises at least 30% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 35% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 40% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 45% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 50% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 55% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 60% of the amino acid sequence selected from Table 1. In some embodiments, the provided modified peptide comprises at least 65% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 70% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 75% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 80% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 85% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 90% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 95% of the amino acid sequence identical to the sequences selected from Table 1. In some embodiments, the provided modified peptide comprises at least 98% of the amino acid sequence identical to the sequences selected from Table 1.

[0050] In some embodiments, this disclosure provides a modified peptide comprising an amino acid sequence having 0 to 10 substitutions, insertions, and / or deletions relative to a sequence selected from Table 1, wherein at least two amino acids comprise side chains linked together (e.g., to form a staple). In some embodiments, the provided modified peptide comprises 0 to 5 substitutions, insertions, and / or deletions relative to a sequence selected from Table 1. In some embodiments, the provided modified peptide comprises 0 to 2 substitutions, insertions, and / or deletions relative to a sequence selected from Table 1. In some embodiments, the provided modified peptide comprises 1 to 5 substitutions, insertions, and / or deletions relative to a sequence selected from Table 1. In some embodiments, the provided modified peptide comprises 1 to 2 substitutions, insertions, and / or deletions relative to a sequence selected from Table 1.

[0051] In some embodiments, when the modified peptide comprises an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is a conserved amino acid substitution. "Conserved amino acid substitution" means a substitution in which an amino acid is replaced by another amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids having basic side chains (…). For example Lysine, arginine, histidine), and amino acids with acidic side chains ( For example Aspartic acid, glutamic acid), and amino acids with uncharged polar side chains ( For example Glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), and amino acids with nonpolar side chains ( For example Alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), and amino acids with β-branched side chains ( For example Threonine, valine, isoleucine), amino acids with aromatic side chains ( For example Tyrosine, phenylalanine, tryptophan, histidine) and amino acids with acidic side chains and their amides ( For example (Aspartic acid, glutamic acid, asparagine, glutamine).

[0052] In some embodiments, when the modified peptide contains an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is methionine to leucine. In some embodiments, when the modified peptide contains an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is threonine to valine. In some embodiments, when the modified peptide contains an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is glutamine to glutamic acid. In some embodiments, when the modified peptide contains an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is phenylalanine to tyrosine. In some embodiments, when the modified peptide contains an amino acid sequence having a substituted amino acid sequence relative to a sequence selected from Table 1, the substitution is arginine to serine.

[0053] In some embodiments, the provided modified peptide comprises the following amino acid sequence: SE-Xaa 1 -NKL-Xaa 2 -EK-Xaa 3 -R-Xaa 4 -Xaa 5 -LRENA (SEQ ID NO: 7) in: Xaa 1 It is either methionine or oroleucine; Xaa 2 It is either phenylalanine or tyrosine; Xaa 3 It could be threonine or valine; Xaa 4 It is arginine or serine; and Xaa 5 It is either glutamine or glutamic acid.

[0054] In some embodiments, the provided modified peptide comprises the following amino acid sequence: SE-Xaa 1 -NKL-Xaa 2 -EKVR-Xaa 4 -Xaa 5 -LRENA (SEQ ID NO: 8) in: Xaa 1 It is either methionine or oroleucine; Xaa 2 It is either phenylalanine or tyrosine; Xaa 4 It is arginine or serine; and Xaa 5 It is either glutamine or glutamic acid.

[0055] In some embodiments, the provided modified peptide comprises the following amino acid sequence: SEMNKL-Xaa 2 -EK-Xaa 3 -R-Xaa 4 -Xaa 5 -LRENA (SEQ ID NO: 9) in: Xaa 2 It is either phenylalanine or tyrosine; Xaa 3 It could be threonine or valine; Xaa 4 It is arginine or serine; and Xaa 5 It is either glutamine or glutamic acid.

[0056] In some embodiments, the provided modified peptide comprises the following amino acid sequence: SE-Xaa 1 -NKL-Xaa 2 -EK-Xaa 3 -R-Xaa 4 -Xaa 5 -LRENA (SEQ ID NO: 10) in: Xaa 1 It is leucine; Xaa 2 It is either phenylalanine or tyrosine; Xaa 3 It could be threonine or valine; Xaa 4 It is arginine or serine; and Xaa 5 It is either glutamine or glutamic acid.

[0057] In some embodiments, the provided modified peptide comprises at least two amino acid side chains linked together (e.g., to form a staple). In some embodiments, the two linked amino acid side chains span one or two turns of the α-helix of the modified peptide. In some embodiments, the two linked amino acid side chains span one turn of the α-helix of the modified peptide. In some embodiments, the two linked amino acid side chains span two turns of the α-helix of the modified peptide.

[0058] In some embodiments, the provided modified peptide comprises at least two amino acid side chains linked together, wherein the two amino acids are separated by: (i) three amino acids (e.g., side chains of the amino acids at positions i and i+4); or (ii) six amino acids (e.g., side chains of the amino acids at positions i and i+7). In some embodiments, the provided modified peptide comprises at least two amino acid side chains linked together, wherein the two amino acids are separated by three amino acids (e.g., side chains of the amino acids at positions i and i+4). In some embodiments, the provided modified peptide comprises at least two amino acid side chains linked together, wherein the two amino acids are separated by six amino acids (e.g., side chains of the amino acids at positions i and i+7).

[0059] In some embodiments, this disclosure provides a modified peptide of formula I':

[0060] Or its pharmaceutically acceptable salt, wherein: Each Xaa independently represents a natural or non-natural amino acid; w, x, and y are independently 0 to 50, including end values; X L1 and X L2 It consists of amino acids that form side chains linked together; [Xaa] x Represents an amino acid sequence containing at least three consecutive amino acids of the following: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to the amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to the amino acid sequence selected from Table 1. [Xaa] w and [Xaa] y Each of these independently represents an amino acid sequence containing one or more consecutive amino acids: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. Choose any one of [Xaa] w The N-terminal amino acid and / or [Xaa] in y The C-terminal amino acid in it is partially conjugated with the one described in this article.

[0061] In some embodiments, this disclosure provides a modified peptide of Formula I:

[0062] I Or its pharmaceutically acceptable salt, wherein: R 1 and R 2 Each is independently hydrogen or optionally substituted C 1-10 Aliphatic groups; R 3 Represents the attached side chain group and is optionally substituted divalent C. 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; Each Xaa independently represents a natural or non-natural amino acid; w and y are each independently between 0 and 50; x is between 3 and 10; and [Xaa] x Represents an amino acid sequence containing at least three consecutive amino acids of the following: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to the amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to the amino acid sequence selected from Table 1. [Xaa] w and [Xaa] y Each of these independently represents an amino acid sequence containing one or more consecutive amino acids: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. Choose any one of [Xaa] w The N-terminal amino acid and / or [Xaa] in y The C-terminal amino acid in it is partially conjugated with the one described in this article.

[0063] In some embodiments, this disclosure provides a modified peptide of formula IA:

[0064] IA Or its pharmaceutically acceptable salts, wherein, alone and in combination, R 3 [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this article.

[0065] In some embodiments, this disclosure provides a modified peptide of formula IB:

[0066] IB Or its pharmaceutically acceptable salts, wherein, alone and in combination, R 3 [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this article.

[0067] In some embodiments, this disclosure provides a modified peptide selected from the following: (I-B1) (I-B2) or (I-B3) Or its pharmaceutically acceptable salts, wherein, alone and in combination, R 3 [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this article.

[0068] In some embodiments, this disclosure provides a modified peptide of formula IC:

[0069] IC Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; p is 2, 3, 4, 5, 6, 7, or 8; and q is 2, 3, 4, 5, or 6.

[0070] In some embodiments, this disclosure provides modified peptides of formula I-Ci:

[0071] I-Ci Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this article.

[0072] As defined in general above, in equation I', X L1 and X L2 For amino acids comprising side chains linked together. In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XI:

[0073] XI In both individual and combined cases, R 1 R 2 and R 3 All are as defined above with respect to Equation I, and are described in the categories and subcategories of this article.

[0074] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XIa:

[0075] XIa In both individual and combined cases, R 3 As defined with respect to Formula I, and described in the categories and subcategories of this document.

[0076] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have a structure selected from the following: (XIb) (XIc), or (XId), In both individual and combined cases, R 3 As defined with respect to Formula I, and described in the categories and subcategories of this document.

[0077] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XII:

[0078] XII Where p is 2, 3, 4, 5, 6, 7 or 8; and q is 2, 3, 4, 5 or 6.

[0079] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XIIa:

[0080] XIIa.

[0081] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XIIb:

[0082] XIIb.

[0083] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XIIc:

[0084] XIIc.

[0085] In some embodiments, X L1 and X L2 (For example, two amino acids containing side chains linked together) have the structure of formula XIId:

[0086] XIId.

[0087] In some embodiments of any of the forms described herein, R 1 It is hydrogen or C 1-10 Aliphatic. In some embodiments, R 1 It is hydrogen. In some embodiments, R is... 1 C is arbitrarily substituted 1-10 Aliphatic (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 C is arbitrarily substituted 1-10 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R1 C is arbitrarily substituted 2-10 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 C is arbitrarily substituted 2-10 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 C 1-10 Alkyl group. In some embodiments, R 1 C 2-10 Alkenyl. In some embodiments, R 1 C 2-10 Alkynyl group. In some embodiments, R 1 C is arbitrarily substituted 1-4 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 C is arbitrarily substituted 2-4 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 C is arbitrarily substituted 2-4 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 C 1-4 Alkyl group. In some embodiments, R 1 C 2-4 Alkenyl. In some embodiments, R 1 C 2-4 Alkynyl group. In some embodiments, R 1 It is a methyl group.

[0088] In some embodiments of any of the forms described herein, R 2 It is hydrogen or C 1-10 Aliphatic. In some embodiments, R 2 It is hydrogen. In some embodiments, R is... 2 C is arbitrarily substituted 1-10 Aliphatic (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 2 C is arbitrarily substituted 1-10 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 2 C is arbitrarily substituted2-10 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 2 C is arbitrarily substituted 2-10 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 2 C 1-10 Alkyl group. In some embodiments, R 2 C 2-10 Alkenyl. In some embodiments, R 2 C 2-10 Alkynyl group. In some embodiments, R 2 C is arbitrarily substituted 1-4 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 2 C is arbitrarily substituted 2-4 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 2 C is arbitrarily substituted 2-4 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 2 C 1-4 Alkyl group. In some embodiments, R 2 C 2-4 Alkenyl. In some embodiments, R 2 C 2-4 Alkynyl group. In some embodiments, R 2 It is a methyl group.

[0089] In some embodiments of any of the forms described herein, R 1 and R 2 Each is hydrogen. In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 1-10 Aliphatic (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 1-10 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2Each independently represents the optionally substituted C 2-10 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 2-10 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently is C 1-10 Aliphatic. In some embodiments, R 1 and R 2 Each independently is C 1-10 Alkyl group. In some embodiments, R 1 and R 2 Each independently is C 2-10 Alkenyl. In some embodiments, R 1 and R 2 Each independently is C 2-10 Alkynyl group. In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 1-4 Aliphatic (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 1-4 Alkyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 2-4 Alkenyl groups (e.g., optionally halogenated, hydroxylated, or C-substituted) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently represents the optionally substituted C 2-4 Alkyne group (e.g., optionally halogenated, hydroxylated, or C-shaped) 1-3 Alkoxy substitution). In some embodiments, R 1 and R 2 Each independently is C 1-4 Aliphatic. In some embodiments, R 1 and R 2 Each independently is C 1-4 Alkyl group. In some embodiments, R 1 and R 2Each independently is C 2-4 Alkenyl. In some embodiments, R 1 and R 2 Each independently is C 2-4 Alkyne group. In some embodiments, R 1 and R 2 Each is a methyl group.

[0090] In some embodiments of any of the forms described herein, R 3 For optional substituted divalent C 7-16 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 For optional substituted divalent C 7-12 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—.

[0091] In some embodiments of any of the forms described herein, R 3 For optional substituted divalent C containing at least one double bond 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 For optional substituted divalent C containing at least one double bond 7-16A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 For optional substituted divalent C containing at least one double bond 7-12 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—.

[0092] In some embodiments of any of the forms described herein, R 3 For optional substituted divalent C 3-20 Hydrocarbon chain. In some embodiments, R 3 For optional substituted divalent C 7-16 Hydrocarbon chain. In some embodiments, R 3 For optional substituted divalent C 7-12 Hydrocarbon chain. In some embodiments, R 3 For optional substituted divalent C containing at least one double bond 3-20 Hydrocarbon chain. In some embodiments, R 3 For optional substituted divalent C containing at least one double bond 7-16 Hydrocarbon chain. In some embodiments, R 3 For optional substituted divalent C containing at least one double bond 7-12 Hydrocarbon chain.

[0093] In some embodiments of any of the forms described herein, R 3 divalent C 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 divalent C 7-16A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 divalent C 7-12 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—.

[0094] In some embodiments of any of the forms described herein, R 3 For divalent C containing at least one double bond 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 For divalent C containing at least one double bond 7-16 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently replaced by —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—. In some embodiments, R 3 For divalent C containing at least one double bond 7-12A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—.

[0095] In some embodiments of any of the forms described herein, R 3 divalent C 3-20 Hydrocarbon chain. In some embodiments, R 3 divalent C 7-16 Hydrocarbon chain. In some embodiments, R 3 divalent C 7-12 Hydrocarbon chain. In some embodiments, R 3 For divalent C containing at least one double bond 3-20 Hydrocarbon chain. In some embodiments, R 3 For divalent C containing at least one double bond 7-16 Hydrocarbon chain. In some embodiments, R 3 For divalent C containing at least one double bond 7-12 Hydrocarbon chain.

[0096] In some embodiments of any of the forms described herein, R 3 C 7-12 Alkenyl group. In some embodiments, R 3 It is C8-olefinic. In some embodiments, R 3 C 11 Alkenyl group. In some embodiments, R 3 -(CH2) 1-7 -CH=CH-(CH2) 1-7 -. In some embodiments, R 3 -(CH2) 3-7 -CH=CH-(CH2) 3-4 -. In some embodiments, R 3 It is -(CH2)6-CH=CH-(CH2)3-. In some embodiments, R 3 It is -(CH2)3-CH=CH-(CH2)3-.

[0097] In some embodiments of any of the forms described herein, R 3 Spanning one or two turns of the α-spiral. In some embodiments, R 3 One revolution across the α-helix. In some embodiments, R 3 It spans two turns of the α-spiral.

[0098] In some embodiments of any of the forms described herein, each R′ is independently hydrogen or optionally substituted C. 1-6 Aliphatic. In some embodiments, R′ is hydrogen. In some embodiments, R′ is optionally substituted C. 1-12 Aliphatic. In some embodiments, R′ is optionally substituted C 1-6 Aliphatic. In some embodiments, R′ is C 1-12 Aliphatic. In some embodiments, R′ is C 1-6 Aliphatic. In some embodiments, R′ is optionally substituted C 1-12 Alkyl group. In some embodiments, R′ is optionally substituted C. 1-6 Alkyl group. In some embodiments, R′ is C 1-12 Alkyl group. In some embodiments, R′ is C 1-6 alkyl.

[0099] In some embodiments of any of the forms described herein, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6. In some embodiments, p is 7. In some embodiments, p is 8.

[0100] In some embodiments of any of the forms described herein, q is 2. In some embodiments, q is 3. In some embodiments, q is 4. In some embodiments, q is 5. In some embodiments, q is 6.

[0101] In some embodiments of any of the forms described herein, p is 5 and q is 3. In some embodiments, p is 2 and q is 3.

[0102] In some embodiments of any of the forms described herein, each Xaa is independently a natural amino acid. In some embodiments, Xaa is a natural amino acid. In some embodiments, Xaa is a non-natural amino acid. In some embodiments, 1 to 5 of Xaa are non-natural amino acids, such as ortholeucine. In some embodiments, 1 to 3 of Xaa are non-natural amino acids, such as ortholeucine. In some embodiments, one of Xaa is a non-natural amino acid, such as ortholeucine. In some embodiments, Xaa is... N -Acyl amino acid (e.g., when Xaa is at the N-terminus of the peptide). In some embodiments, Xaa is an α-aminoamide (e.g., when Xaa is at the C-terminus of the peptide).

[0103] In some embodiments of any of the forms described herein, w is an integer from 2 to 50. In some embodiments, w is an integer from 2 to 25. In some embodiments, w is an integer from 2 to 10. In some embodiments, w is 7. In some embodiments, w is 11.

[0104] In some embodiments of any of the forms described herein, [Xaa] w An amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least seven consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w It does not exist (i.e., when w is 0).

[0105] In some embodiments of any of the forms described herein, [Xaa] w An amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] wAn amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least seven consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0106] In some embodiments of any of the forms described herein, [Xaa] w An amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] wAn amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least seven consecutive amino acids containing: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0107] In some embodiments of any of the forms described herein, [Xaa] w This represents an amino acid sequence comprising one or more consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] w This represents an amino acid sequence comprising at least two consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] w This represents an amino acid sequence comprising at least three consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] w This represents an amino acid sequence comprising at least five consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] w This represents an amino acid sequence containing at least 7 consecutive amino acids selected from the amino acid sequences in Table 1.

[0108] In some embodiments of any of the forms described herein, [Xaa] w This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 30% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] w This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 70% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] w An amino acid sequence representing at least three consecutive amino acids that are at least 90% identical to the amino acid sequences selected from Table 1.

[0109] In some embodiments of any of the forms described herein, [Xaa] w This represents an amino acid sequence comprising at least three consecutive amino acids having one to ten substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] wThis represents an amino acid sequence comprising at least three consecutive amino acids having one to five substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] w It represents an amino acid sequence containing at least three consecutive amino acids having one to three substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1.

[0110] In some embodiments of any of the forms described herein, x is an integer from 3 to 50. In some embodiments, x is an integer from 3 to 25. In some embodiments, x is an integer from 3 to 10. In some embodiments, x is an integer from 3 to 5. In some embodiments, x is an integer from 5 to 10. In some embodiments, x is 3 or 6. In some embodiments, x is 3. In some embodiments, x is 6.

[0111] In some embodiments of any of the forms described herein, [Xaa] x An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least six consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least seven consecutive amino acids containing: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0112] In some embodiments of any of the forms described herein, [Xaa] xAn amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least six consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least seven consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0113] In some embodiments of any of the forms described herein, [Xaa] x An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least six consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] xAn amino acid sequence representing at least seven consecutive amino acids containing: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0114] In some embodiments of any of the forms described herein, [Xaa] x This represents an amino acid sequence comprising at least three consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] x This represents an amino acid sequence comprising at least five consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] x This represents an amino acid sequence comprising at least six consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] x This represents an amino acid sequence containing at least 7 consecutive amino acids selected from the amino acid sequences in Table 1.

[0115] In some embodiments of any of the forms described herein, [Xaa] x This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 30% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] x This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 70% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] x An amino acid sequence representing at least three consecutive amino acids that are at least 90% identical to the amino acid sequences selected from Table 1.

[0116] In some embodiments of any of the forms described herein, [Xaa] x This represents an amino acid sequence comprising at least three consecutive amino acids having one to ten substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] x This represents an amino acid sequence comprising at least three consecutive amino acids having one to five substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] x It represents an amino acid sequence containing at least three consecutive amino acids having one to three substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1.

[0117] In some embodiments of any of the forms described herein, y is an integer from 2 to 50. In some embodiments, y is an integer from 2 to 25. In some embodiments, y is an integer from 2 to 10. In some embodiments, y is an integer from 0 to 5. In some embodiments, y is an integer from 2 to 5. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, y is 3. In some embodiments, y is 4.

[0118] In some embodiments of any of the forms described herein, [Xaa] y An amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least seven consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y It does not exist (i.e., when y is 0).

[0119] In some embodiments of any of the forms described herein, [Xaa] yAn amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least seven consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 70% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0120] In some embodiments of any of the forms described herein, [Xaa] y An amino acid sequence representing one or more consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least two consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] yAn amino acid sequence representing at least three consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least five consecutive amino acids comprising: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least seven consecutive amino acids containing: an amino acid sequence selected from Table 1; or (i) at least 90% identical to an amino acid sequence selected from Table 1 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1.

[0121] In some embodiments of any of the forms described herein, [Xaa] y This represents an amino acid sequence comprising one or more consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence comprising at least two consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence comprising at least three consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence comprising at least five consecutive amino acids selected from the amino acid sequences in Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence containing at least 7 consecutive amino acids selected from the amino acid sequences in Table 1.

[0122] In some embodiments of any of the forms described herein, [Xaa] y This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 30% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence comprising at least three consecutive amino acids that are at least 70% identical to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] y An amino acid sequence representing at least three consecutive amino acids that are at least 90% identical to the amino acid sequences selected from Table 1.

[0123] In some embodiments of any of the forms described herein, [Xaa] yThis represents an amino acid sequence comprising at least three consecutive amino acids having one to ten substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] y This represents an amino acid sequence comprising at least three consecutive amino acids having one to five substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1. In some embodiments, [Xaa] y It represents an amino acid sequence containing at least three consecutive amino acids having one to three substitutions, insertions, and / or deletions relative to the amino acid sequences selected from Table 1.

[0124] In some embodiments of any of the forms described herein, [Xaa] w The N-terminal amino acid in [Xaa] is partially conjugated as described herein. In some embodiments, [Xaa] y The C-terminal amino acid in [Xaa] is partially conjugated as described herein. In some embodiments, [Xaa] w The N-terminal amino acid and [Xaa] in y The C-terminal amino acid in it is partially conjugated with the one described in this article.

[0125] In some embodiments of any of the forms described herein, [Xaa] w Selected from Table 2.

[0126] Table 2.

[0127] In some embodiments of any of the forms described herein, [Xaa] x Selected from Table 3.

[0128] Table 3.

[0129] In some embodiments of any of the forms described herein, [Xaa] y Selected from Table 4.

[0130] Table 4.

[0131] In some embodiments of any of the forms described herein, [Xaa] w [Xaa] x and [Xaa] y Selected from Table 5.

[0132] Table 5.

[0133] B* = Leucine It should be understood that [Xaa] in Tables 2 to 5 w [Xaa] x and [Xaa] y The amino acid sequence may be modified to include additional amino acids at the N and / or C ends (e.g., adding 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids), and / or to have N and / or C-terminal deletions (e.g., deleting 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids). In some cases, the sequences in Tables 2 to 5 include one or more variants. In some embodiments, the amino acid sequence has 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, or 2 substitutions, insertions, and / or deletions relative to the sequences in Tables 2 to 5. In some embodiments, the [Xaa] described herein w [Xaa] x and [Xaa] y The amino acid sequence (e.g., the sequences in Tables 2 to 5) may also contain one or more (e.g., 1, 2, 3, 4, or 5) amino acid substitutions (relative to the amino acid sequence shown in any of the sequences in Tables 2 to 5), for example, one or more ( For example (1, 2, 3, 4 or 5) Conserved and / or non-conserved amino acid substitutions.

[0134] In some embodiments of any of the forms described herein, the length of the modified peptide described herein is at most 50 amino acids (i.e., w+x+y+2 ≤ 50). In some embodiments, the length of the modified peptide described herein is at most 45 amino acids (i.e., w+x+y+2 ≤ 45). In some embodiments, the length of the modified peptide described herein is at most 30 amino acids (i.e., w+x+y+2 ≤ 30). In some embodiments, the length of the modified peptide described herein is 22 amino acids (i.e., w+x+y+2 = 22). In some embodiments, the length of the modified peptide described herein is 22 amino acids (i.e., w+x+y+2 = 18).

[0135] In some embodiments, the modified peptides described herein comprise peptide pinning. “Peptide pinning” is a term coined by synthetic methods, in which two amino acid side chains are covalently linked (e.g., “pinned together”). For example, two olefinic side chains present in a peptide chain ( For exampleCrosslinkable side chains can be covalently linked using ring-closed metathesis (RCM) reactions. For example "Stapled together" to form cross-linked rings (see, For example Blackwell et al., J. Org. Chem., 66: 5291-5302, 2001; Angew et al., Chem. Int. Ed. 37:3281, 1994. Peptide stabilization can confer structural stability (see Blackwell et al., J. Org. Chem., 66: 5291-5302, 2001; Angew et al., Chem. Int. Ed. 37:3281, 1994). ... example like Walensky J. Med. Chem ., 57:6275-6288 (2014)). In some cases, pegging is hydrocarbon pegging.

[0136] In some cases, the pinning described herein is lactam pinning; UV-cycloaddition pinning; oxime pinning; thioether pinning; double-click pinning; dilactam pinning; diarylyl pinning; or any combination of two or more thereof. Stable peptides as described herein include pinned peptides as well as peptides containing multiple pins or any other chemical strategy for structural reinforcement (see [link]). example like Balaram P. Cur. Opin. Struct. Biol 1992;2:845; Kemp DS et al. , J. Am. Chem. Soc. 1996;118:4240; Orner BP et al., J. Am. Chem. Soc 2001;123:5382; Chin JW et al., Int. Ed. 2001;40:3806; Chapman RN et al., J. Am. Chem. Soc 2004;126:12252; Horne WS et al., Chem., Int. Ed 2008;47:2853; Madden et al., Chem Commun (Camb). October 7, 2009; (37): 5588–5590; Lau et al., Chem. Soc. Rev ., 2015, 44: 91-102; and Gunnoo et al., Org. Biomol. Chem ., 2016,14:8002-8013).

[0137] In some embodiments, the stapled peptide is structurally stable. A peptide is "structurally stable" because it retains its native secondary structure. For example, stapled peptides tend to have an α-helical secondary structure to maintain their native α-helical conformation. This secondary structure increases the peptide's resistance to proteolytic cleavage and heat, and can increase target binding affinity, hydrophobicity, plasma membrane binding, and / or cell permeability. Therefore, the modified (e.g., stapled) peptides described herein have improved biological activity and pharmacology compared to their corresponding unmodified (e.g., non-stapled (uncrosslinked)) peptides.

[0138] Non-limiting examples of non-natural amino acids that can be used as stapled amino acids are: (R)-2-(2'-propenyl)alanine; (R)-2-(4′-pentenyl)alanine; (R)-α-(7′-octenyl)alanine; (S)-α-(2′-propenyl)alanine; (S)-α-(4′-pentenyl)alanine; (S)-2-(7′-octenyl)alanine; α,α-bis(4′-pentenyl)glycine; and α,α-bis(7′-octenyl)glycine.

[0139] In some embodiments, the tethering occurs between two amino acids separated by, for example, six amino acids. In some embodiments, the amino acids forming the tether are at positions i and i+7. For example, where the peptide has the sequence ...X1, X2, X3, X4, X5, X6, X7, X8, X9, X10 ..., the tethering can be between X2 and X9 or X3 and X10, etc. (i and i+7). Where the peptide has the sequence ...X1, X2, X3, X4, X5, X6, X7, X8, X9 ..., the tethering can be between X2 and X6, X3 and X7 or X4 and X8, etc. (i and i+4). Further descriptions of the preparation and use of hydrocarbon-tethered peptides can be found in […]. ,For example In U.S. Patent Publications 2012 / 0172285, 2010 / 0286057 and 2005 / 0250680.

[0140] In addition, other methods for performing different types of fastening are well known in the art and can be used in conjunction with the modified peptides described herein. See , For example Lactam stapling: Shepherd et al. , J. Am. Chem. Soc .,127:2974–2983 (2005); UV-cycloaddition stapling: Madden et al. , Bioorg. Med. Chem.Lett., 21:1472–1475 (2011); Disulfide stapling: Jackson et al. A m. Chem. Soc. ,113:9391–9392 (1991); Oxime stapling: Haney et al. , Chem. Commun ., 47:10915–10917(2011); Thioether stapling: Brunel and Dawson, Chem. Commun ., 552–2554 (2005); Photoswitchable stapling: JR Kumita et al. , Proc. Natl. Acad. Sci. US A .,97:3803–3808 (2000); Double-click stapling: Lau et al. , Chem. Sci ., 5:1804–1809(2014); Bis-lactam stapling: JC Phelan et al. , J. Am. Chem. Soc ., 119:455–460(1997); and Bis-arylation stapling: AM Spokoyny et al. , J. Am. Chem. Soc ., 135:5946–5949 (2013)).

[0141] Conjugate In some embodiments, the modified peptide provided herein is conjugated to one or more portions. In some embodiments, the conjugated portion is a portion that imparts a useful characteristic to the modified peptide, such as improved pharmacokinetic or pharmacodynamic properties. In some embodiments, the conjugated portion is or comprises a lipid (e.g., cholesterol). In some embodiments, the conjugated portion is or comprises a carbohydrate. In some embodiments, the conjugated portion is or comprises a peptide (e.g., the second modified peptide described herein). In some embodiments, the conjugated portion is attached to the modified peptide via a linker (e.g., a linker comprising polyethylene glycol (PEG)).

[0142] In some embodiments, a portion is conjugated at or near the N-terminus of the peptide. For example, in some embodiments, the portion is conjugated to the α-amino group of the N-terminal amino acid. In some embodiments, a portion is conjugated at or near the C-terminus of the peptide. For example, in some embodiments, a lysine residue is attached to the C-terminus of the peptide and a portion is conjugated to the lysine side chain (e.g., via an amino group).

[0143] In some embodiments, the provided modified peptide comprises: –(connector)-(lipid moiety). In some embodiments, the connector comprises PEG, a peptide (e.g., one or more amino acids), or a combination thereof. In some embodiments, the connector comprises a peptide. In some embodiments, the connector comprises one or more PEG units. In some embodiments, the connector comprises PEG units linked by an amide bond. In some embodiments, the amino acid at the C-terminus of the provided modified peptide is conjugated to the lipid moiety via a connector group, i.e., –(connector)-(lipid moiety). In some embodiments, the amino acid at the N-terminus of the provided modified peptide is conjugated to the lipid moiety via a connector group, i.e., –(connector)-(lipid moiety).

[0144] In some embodiments, the linker comprises PEG. In some embodiments, the linker is a peptide comprising 3 to 20 amino acids. Linkers considered herein include those disclosed in the following literature: Chen X. et al. Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev. Oct 2013;65(10):1357-69 and Zhou, J. et al. A highly potent and stable pan-coronavirus fusion inhibitor as a candidate prophylactic and therapeutic for COVID-19 and other coronavirus diseases, Acta Pharm. Sinica B; 2022;12(4): 1652-1661.

[0145] In some embodiments, the lipid portion is C5-C. 100 The lipid portion is an aliphatic lipid. In some embodiments, the lipid portion is a fatty acid, phospholipid, sphingolipid, or sterol. In some embodiments, the fatty acid is palmitic acid or stearic acid. In some embodiments, the phospholipid is phosphatidylcholine or phosphatidylethanolamine. In some embodiments, the sphingolipid is sphingosine or ceramide. In some embodiments, the sterol is cholesterol, mercaptocholesterol, sitosterol, ergosterol, or tocopherol. In some embodiments, the lipid portion is cholesterol.

[0146] In some embodiments, this disclosure provides a modified peptide of formula II':

[0147] II' Or its pharmaceutically acceptable salts, wherein, alone and in combination, R1 R 2 R 3 [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O- or -[O-CH2CH2] n Replacement, where n is from 1 to 20; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; and R 6 For example, conjugated parts (e.g., lipids, carbohydrates, or peptides).

[0148] In some embodiments, this disclosure provides a modified peptide of formula II”:

[0149] II” Or its pharmaceutically acceptable salts, wherein, alone and in combination, R 1 R 2 R 3 [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O- or -[O-CH2CH2] nReplacement, where n is from 1 to 20; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; and R 6 For example, conjugated parts (e.g., lipids, carbohydrates, or peptides).

[0150] In some embodiments, this disclosure provides a modified peptide of Formula II:

[0151] II Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O- or -[O-CH2CH2] n Replacement, where n is from 1 to 20; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide). p is 2, 3, 4, 5, 6, 7, or 8; and q can be 2, 3, 4, 5 or 6.

[0152] In some embodiments, this disclosure provides a modified peptide of formula IIa:

[0153] IIa Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 4C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O- or -[O-CH2CH2] n Replacement, where n is from 1 to 20; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; and R 6 For example, conjugated parts (e.g., lipids, carbohydrates, or peptides).

[0154] In some embodiments, this disclosure provides a modified peptide of formula IIb:

[0155] IIb Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2) 0-6 -R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide). m is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16; p is 2, 3, 4, 5, 6, 7, or 8; and q can be 2, 3, 4, 5 or 6.

[0156] In some embodiments, this disclosure provides a modified peptide of formula IIc:

[0157] IIc Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2) 0-6 -R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide); and m is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0158] In some embodiments, this disclosure provides a modified peptide of formula IVc:

[0159] IVc Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2) 0-6 -R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide); and m is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0160] In some embodiments, this disclosure provides a modified peptide of formula Va:

[0161] Va Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 8 -C(O)(C 1-4 alkyl); R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2) 0-6 -R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide); and m is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0162] In some embodiments, this disclosure provides a modified peptide of formula Vb:

[0163] Vb Or its pharmaceutically acceptable salts, wherein, alone and in combination, [Xaa] w [Xaa] x and [Xaa] y All are as defined above with respect to Equation I, and are described in the categories and subcategories of this document; and R 8 -C(O)(C 1-4 alkyl); R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2) 0-6 -R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 For example, a conjugated portion (e.g., lipid, carbohydrate, or peptide); and m is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0164] In some embodiments of any of the forms described herein, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O-, or -[O-CH2CH2] n Instead, where n is 1 to 20. In some embodiments, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 6-30 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O-, or -[O-CH2CH2] n Instead, where n is 1 to 20. In some embodiments, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 3-20 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O-, or -[O-CH2CH2] n Replacement, where n is from 1 to 20.

[0165] In some embodiments of any of the forms described herein, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -N(R′)C(O)-, -C(O)N(R′)-, or -[O-CH2CH2]. n Instead, where n is 1 to 20. In some embodiments, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 6-30A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -N(R′)C(O)-, -C(O)N(R′)-, or -[O-CH2CH2]. n Instead, where n is 1 to 20. In some embodiments, R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 3-20 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -N(R′)C(O)-, -C(O)N(R′)-, or -[O-CH2CH2]. n Replacement, where n is from 1 to 20.

[0166] In some embodiments of any of the forms described herein, R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-6 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-6 (alkylene)-R 6 ))-C(O)-NH2. In some embodiments, R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-3 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-2 (alkylene)-R 6 ))-C(O)-NH2. In some embodiments, R 4 is -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] n -N(R 5 )C(O)-CH2-R 6 ))-C(O)-NH2. In some embodiments, R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-3 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-2 (alkylene)-R 6 ))-C(O)-NH2. In some embodiments, R 4 is -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] n -N(R 5 )C(O)-CH2-R 6))-C(O)-NH2. In some embodiments, R 4 -N(H)-C(H)(CH2)4N(H)(C(O)-(C 2-3 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-2 (alkylene)-R 6 ))-C(O)-NH2. In some embodiments, R 4 is -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] n -N(R 5 )C(O)-R 6 ))-C(O)-NH2. In some embodiments, R 4 is -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] n -N(R 5 )C(O)-R 6 ))-C(O)-NH2.

[0167] In some embodiments of any of the forms described herein, R 4 It is a divalent peptide. In some embodiments, R... 4 It contains amino acid residues (e.g., lysine).

[0168] In some embodiments of any of the forms described herein, R 5 It is hydrogen. In some embodiments, R is... 5 C 1-4 Alkyl group. In some embodiments, R 5 It is a methyl group.

[0169] In some embodiments of any of the forms described herein, R 6 It is a lipid. In some embodiments, R 6 R is a carbohydrate. In some embodiments, R 6 It is a peptide.

[0170] In some embodiments of any of the forms described herein, R 6 C5-C is optionally replaced 100 Aliphatic lipid moiety. In some embodiments, R 6 It is a fatty acid, phospholipid, sphingolipid, or sterol. In some embodiments, R 6 It is palmitic acid or stearic acid. In some embodiments, R 6 It is phosphatidylcholine or phosphatidylethanolamine. In some embodiments, R 6It is sphingosine or ceramide. In some embodiments, R 6 It can be cholesterol, mercaptocholesterol, sitosterol, ergosterol, or tocopherol.

[0171] In some embodiments of any of the forms described herein, R 6 One of the following: , , , or , Each of these may optionally be substituted (e.g., halogenated, C-substituted) 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy). In some embodiments, R 6 for , , , or .

[0172] In some embodiments, R 6 for It can be optionally substituted (e.g., halogenated, C-substituted). 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy). In some embodiments, R 6 for It can optionally be substituted (e.g., halogenated, C-substituted) 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy). In some embodiments, R 6 for It can optionally be substituted (e.g., halogenated, C-substituted) 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy). In some embodiments, R 6 for It can optionally be substituted (e.g., halogenated, C-substituted) 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy). In some embodiments, R 6 for It can optionally be substituted (e.g., halogenated, C-substituted) 1-3 Alkyl, hydroxyl or C 1-3 Alkoxy group).

[0173] In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for In some embodiments, R 6 for .

[0174] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0175] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0176] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0177] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0178] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0179] In some embodiments of any of the forms described herein, -R 4 -R 6 for: .

[0180] In some embodiments of any of the forms described herein, R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-(CH2)-R 6 In some embodiments, R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] 12 -N(R 5 )C(O)-(CH2)-R 6 In some embodiments, R 9 -C(O)-(CH2CH2)-[O-CH2CH2]8-N(R) 5 )C(O)-(CH2)-R 6 In some embodiments, R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] m -N(R 5 )C(O)-R 6 In some embodiments, R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] 12 -N(R 5 )C(O)-R 6 In some embodiments, R 9 -C(O)-(CH2CH2)-[O-CH2CH2]8-N(R) 5 )C(O)-R 6 .

[0181] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0182] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0183] In some embodiments of any of the forms described herein, -R 9 -R 6for: .

[0184] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0185] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0186] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0187] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0188] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0189] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0190] In some embodiments of any of the forms described herein, -R 9 -R 6 for: .

[0191] In some embodiments of any of the forms described herein, m is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. In some embodiments, m is 6, 7, 8, 9, 10, 11, 12, 13, or 14. In some embodiments, m is 8. In some embodiments, m is 12.

[0192] In some embodiments of any of the forms described herein, n is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. In some embodiments, n is 6, 7, 8, 9, 10, 11, 12, 13, or 14. In some embodiments, n is 8. In some embodiments, n is 12.

[0193] In some embodiments of any of the forms described herein, R 8 -C(O)(C 1-2 Alkyl). In some embodiments, R 8 It is -C(O)CH3.

[0194] In some embodiments, this disclosure provides modified peptides selected from Table 6.

[0195] Table 6.

[0196] in R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] 12 -N(H)C(O)-R 6 ;and R 6 for

[0197]

[0198] B* = Leucine In some embodiments, the provided modified peptide is a peptide further comprising a –(connector)-(peptide moiety). In some embodiments, the connector comprises PEG, a peptide (e.g., one or more amino acids), or a combination thereof. In some embodiments, the connector comprises a peptide. In some embodiments, the connector comprises one or more PEG units. In some embodiments, the connector comprises PEG units linked by an amide bond. In some embodiments, an amino acid at the C-terminus of the provided modified peptide is conjugated to a second peptide moiety via a connector group, i.e., –(connector)-(peptide moiety). In some embodiments, the second peptide moiety is further conjugated to a lipid moiety, i.e., –(connector)-(second peptide moiety)-(connector)-(lipid moiety). In some such embodiments, the lipid moiety is the lipid moiety described herein. In some embodiments, the second peptide moiety has the same or different sequence as the modified peptide. In some embodiments, the second peptide moiety comprises a sequence of amino acids capable of binding to a portion of influenza HA.

[0199] In some embodiments, the provided modified peptide is part of a peptide conjugate of formula XX: (Modified peptide) – (First linker) – (Second peptide moiety) – (Second linker) – (Lipid moiety) XX Or its pharmaceutically acceptable salt, wherein: The modified peptides are those described herein; The first connector is a divalent linear or branched, saturated or unsaturated, optionally substituted C. 1-30 A hydrocarbon chain in which 1 to 10 methylene units are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O-, or -[O-CH2CH2] n Replacement, where n is from 1 to 20; The second peptide portion is a peptide with 10 to 30 amino acids; The second connector is as described in this article, R. 4 Or R 9 ;and The lipid portion is R as described in this article. 6 .

[0200] In some embodiments, the provided modified peptide is part of a peptide conjugate of formula XXI:

[0201] XXI Or its pharmaceutically acceptable salt, wherein: The modified peptides are those described herein; The second peptide portion is a peptide with 10 to 30 amino acids; R 4 As described in this article; and R 6 As described in this article.

[0202] In some embodiments of Formulas XX and XXI, the second peptide portion comprises the following sequence: GTYDHDVYRDEALNNRFQI (SEQ ID NO: 195). In some embodiments, the second peptide portion comprises a sequence that is 30% identical to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence that is 70% identical to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence that is 90% identical to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence that is 95% identical to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence that is 98% identical to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence with 0 to 10 substitutions, insertions, and / or deletions relative to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence with 0 to 5 substitutions, insertions, and / or deletions relative to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises a sequence with 0 to 2 substitutions, insertions, and / or deletions relative to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises sequences having 1 to 5 substitutions, insertions, and / or deletions relative to SEQ ID NO: 195. In some embodiments, the second peptide portion comprises sequences having 1 to 2 substitutions, insertions, and / or deletions relative to SEQ ID NO: 195.

[0203] polymer This disclosure also provides multimeric constructs comprising more than one modified peptide as described herein. In some embodiments, the multimeric construct comprises two modified peptides as described herein.

[0204] In some embodiments, the polymeric construct includes the formula XXX:

[0205] XXX Or its pharmaceutically acceptable salt, wherein: The first modified peptide and the second modified peptide are each independently the modified peptides described herein; The connector is a connecting group; and R 6 As defined in this article.

[0206] In some embodiments, the first modified peptide and the second modified peptide contain the same amino acid sequence. In some embodiments, the first modified peptide and the second modified peptide contain different amino acid sequences.

[0207] Composition This disclosure also provides compositions comprising the modified peptides described herein and one or more other components. In some embodiments, the provided compositions comprise and / or deliver the modified peptides described herein.

[0208] In some embodiments, the provided composition is a pharmaceutical composition comprising and / or delivering the modified peptides provided herein and further comprising a pharmaceutically acceptable carrier.

[0209] The provided pharmaceutical compositions can be specifically formulated for administration in solid or liquid form, including those suitable for: (1) oral administration, such as oral enemas (aqueous or non-aqueous solutions or suspensions), tablets ( For example (1) Targeted oral, sublingual and systemic absorption, pills, powders, granules, pastes for tongue application; (2) Parenteral administration, for example by subcutaneous, intramuscular, intravenous or epidural injection, for example as a sterile solution or suspension or sustained-release formulation; (3) Topical application, for example as a cream, ointment or sustained-release patch or spray applied to the skin; (4) Intravaginal or rectal application, for example as a vaginal suppository, cream or foam; (5) Sublingual application; (6) Ocular application; (7) Transdermal application; (8) Nasal application; or (9) Inhalation application (for example, as an aerosol or used with a nebulizer).

[0210] In some embodiments, the provided pharmaceutical composition comprises the modified peptides described herein and one or more fillers, disintegrants, lubricants, flow aids, anti-adhesives, and / or antistatic agents. The provided pharmaceutical composition may be in various forms, including oral dosage forms, topical creams, topical patches, iontophoresis forms, suppositories, nasal sprays and / or inhalers, eye drops, intraocular injection forms, reservoir forms, and injectable and infusionable solutions. Methods for preparing the pharmaceutical composition are well known in the art.

[0211] use In some embodiments, this disclosure provides methods for treating and / or preventing influenza virus infection using any of the peptides or pharmaceutical compositions described herein.

[0212] In some embodiments, this disclosure provides a method for inhibiting the fusion of influenza virus with host cells in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a peptide comprising an amino acid sequence of influenza hemagglutinin (HA) protein or a variant thereof.

[0213] In some embodiments, this disclosure provides a method for inhibiting conformational changes in influenza hemagglutinin (HA) protein in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a peptide comprising the amino acid sequence of influenza hemagglutinin (HA) protein or a variant thereof. In some embodiments, the inhibited conformational change in influenza hemagglutinin (HA) protein is a foldback of the C-terminal helix of the native influenza HA2 protein (as described in Example 1).

[0214] In some embodiments, this disclosure provides a method for preventing and / or treating influenza virus infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a peptide comprising an amino acid sequence of influenza hemagglutinin (HA) protein or a variant thereof.

[0215] In some embodiments, this disclosure provides a method for preventing influenza virus infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a peptide comprising an amino acid sequence of influenza hemagglutinin (HA) protein or a variant thereof.

[0216] In some embodiments, this disclosure provides a method for treating an influenza virus infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a peptide comprising an amino acid sequence of an influenza hemagglutinin (HA) protein or a variant thereof.

[0217] In some embodiments, a peptide comprising an amino acid sequence of influenza hemagglutinin (HA) protein or a variant thereof inhibits the fusion of influenza virus particles with the endosome membrane in a host cell. In some embodiments, the peptide comprises an amino acid sequence of the HA2 domain of influenza HA. ​​In some embodiments, the peptide comprises an amino acid sequence of a C-terminal helix of the HA2 domain of influenza HA. ​​In some embodiments, the peptide comprises at least five consecutive amino acids selected from the amino acid sequences in Table 1, or (i) is at least 30% identical to the amino acid sequences selected from Table 1; and / or (ii) has one to ten substituted, inserted, and / or deleted amino acid sequences relative to the amino acid sequences selected from Table 1. In some embodiments, the peptide is any of the peptides disclosed herein.

[0218] In some embodiments, this document provides a method for inhibiting the fusion of influenza virus particles with host cells in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein. In some embodiments, the modified peptide inhibits the fusion of more than one (e.g., two, three, four, five, six, seven, eight, nine, ten, or more than ten) influenza A virus subtypes with host cells. In some embodiments, the modified peptide inhibits the fusion of both influenza A and influenza B viruses with host cells.

[0219] In some embodiments, this document provides a method for inhibiting conformational changes in influenza hemagglutinin (HA) proteins in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein. In some embodiments, the inhibited conformational change in the influenza hemagglutinin (HA) protein is a foldback of the C-terminal helix of the native influenza HA2 (as described in Example 1). In some embodiments, the modified peptide or a pharmaceutically acceptable salt thereof inhibits conformational changes in influenza hemagglutinin (HA) proteins of more than one (e.g., two, three, four, five, six, seven, eight, nine, ten, or more than ten) influenza A virus subtypes. In some embodiments, the modified peptide or a pharmaceutically acceptable salt thereof inhibits conformational changes in influenza hemagglutinin (HA) proteins of both influenza A and influenza B viruses.

[0220] In some embodiments, this document provides methods for preventing and / or treating influenza virus infection in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein. In some embodiments, this document provides methods for preventing influenza virus infection in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein. In some embodiments, this document provides methods for treating influenza virus infection in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein.

[0221] In some embodiments, this document provides a method for preventing and / or treating influenza virus infection in a subject of need, the method comprising administering to the subject a therapeutically effective amount of the modified peptide disclosed herein, wherein the modified peptide treats or prevents influenza virus infection caused by more than one (e.g., two, three, four, five, six, seven, eight, nine, ten, or more than ten) influenza A virus subtypes. In some embodiments, the modified peptide or a pharmaceutically acceptable salt thereof treats or prevents influenza virus infection caused by both influenza A and influenza B viruses.

[0222] In some embodiments, influenza A (HA) is derived from influenza A virus. In some embodiments, influenza A is subtypes H10N4, H10N5, H10N7, H10N8, H10N9, H11N1, H11N13, H11N2, H11N4, H11N6, H11N8, H11N9, H12N1, H12N4, H12N5, H12N8, H13N2, H13N3, and H13N4. N6, subtype H13N7, subtype H14N5, subtype H14N6, subtype H15N8, subtype H15N9, subtype H16N3, subtype H1N1, subtype H1N2, subtype H1N3, subtype H1N6, subtype H1N9, subtype H2N1, subtype H2N2, subtype H2N3, subtype H2N5, subtype H2N7, subtype H2N8, subtype H2N9, subtype H3N1, subtype H3N2, subtype H3N3, subtype H3N4, subtype Type H3N5, Subtype H3N6, Subtype H3N8, Subtype H3N9, Subtype H4N1, Subtype H4N2, Subtype H4N3, Subtype H4N4, Subtype H4N5, Subtype H4N6, Subtype H4N8, Subtype H4N9, Subtype H5N1, Subtype H5N2, Subtype H5N3, Subtype H5N4, Subtype H5N6, Subtype H5N7, Subtype H5N8, Subtype H5N9, Subtype H6N1, Subtype H6N2, Subtype H6N3, Subtype H6 The subtypes are: H4, H6N5, H6N6, H6N7, H6N8, H6N9, H7N1, H7N2, H7N3, H7N4, H7N5, H7N7, H7N8, H7N9, H8N4, H8N5, H9N1, H9N2, H9N3, H9N5, H9N6, H9N7, H9N8, or H9N9. In some embodiments, influenza A is subtype H1N1 or H3N2. In some embodiments, influenza HA originates from influenza B virus.

[0223] In some embodiments, the subject has an influenza infection or is at risk of contracting an influenza infection. In some embodiments, the influenza infection is from an influenza A virus. In some embodiments, the influenza A virus is subtype H10N4, subtype H10N5, subtype H10N7, subtype H10N8, subtype H10N9, subtype H11N1, subtype H11N13, subtype H11N2, subtype H11N4, subtype H11N6, subtype H11N8, subtype H11N9, subtype H12N1, subtype H12N4, subtype H12N5, subtype H12N8, subtype H13N2, subtype H13N3, subtype H1 Subtypes H13N6, H14N5, H14N6, H15N8, H15N9, H16N3, H1N1, H1N2, H1N3, H1N6, H1N9, H2N1, H2N2, H2N3, H2N5, H2N7, H2N8, H2N9, H3N1, H3N2, H3N3, H3N4 Subtypes H3N5, H3N6, H3N8, H3N9, H4N1, H4N2, H4N3, H4N4, H4N5, H4N6, H4N8, H4N9, H5N1, H5N2, H5N3, H5N4, H5N6, H5N7, H5N8, H5N9, H6N1, H6N2, H6N3, and H... The influenza A virus includes subtypes H6N4, H6N5, H6N6, H6N7, H6N8, H6N9, H7N1, H7N2, H7N3, H7N4, H7N5, H7N7, H7N8, H7N9, H8N4, H8N5, H9N1, H9N2, H9N3, H9N5, H9N6, H9N7, H9N8, or H9N9. In some embodiments, the influenza A virus is subtype H1N1 or H3N2. In some embodiments, the influenza infection originates from influenza B virus.

[0224] In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal, such as a non-primate ( For example (cattle, pigs, horses, cats, dogs, rats, etc.) or primates ( For example (monkeys or humans). In some embodiments, the subjects are domesticated animals ( For example (dog or cat). In some embodiments, the subject is a human.

[0225] In some embodiments, the subject was infected with influenza virus ( For example(Influenza A or Influenza B). In some embodiments, the subject is infected with the influenza virus ( For example They are at risk of contracting influenza A or influenza B.

[0226] The modified peptides provided can be administered in any suitable manner. In some embodiments, the peptides are administered to the subject orally, intranasally, intravenously, subcutaneously, intramuscularly, or topically (including through the skin, nose, sinuses, eyes, oropharynx, respiratory tree, and lungs, e.g., via inhalation). In some embodiments, the peptides are administered via local respiratory administration (including application to the nasal mucosa, sinus mucosa, oropharyngeal mucosa, or respiratory tree, including the lungs). In some embodiments, the peptides are administered by local application to the skin or eyes.

[0227] The effective amount of the modified peptide provided can be administered in one or more applications, by application, or by dosage. Therapeutic effective amount of therapeutic peptide ( Right now The effective dose depends on the selected therapeutic peptide. The provided composition (e.g., comprising one or more modified peptides as described herein) can be administered once or more daily to once or more weekly. Treating a subject with a therapeutically effective amount of the therapeutic peptide described herein can comprise a single treatment or a series of treatments. For example, an effective amount can be administered at least once.

[0228] In some embodiments, the provided modified peptides (including the conjugates and polymeric constructs described herein) exhibit specific effects in one or more assays described herein. In some embodiments, the provided modified peptides exhibit EC50 values ​​of less than 5 µM in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit EC50 values ​​of less than 2 µM in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit EC50 values ​​of less than 1 µM in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit a selectivity index (SI) greater than 1.5 in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit an SI greater than 2 in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit an SI greater than 4 in the assays of Examples 3 to 5. In some embodiments, the provided modified peptides exhibit an SI greater than 5 in the assays of Examples 3 to 5.

[0229] Method for preparing the provided peptide This disclosure also provides a method for preparing the modified peptides disclosed herein, the method comprising: (a) providing a peptide comprising the amino acid sequence shown in Table 1 or a variant thereof, optionally wherein the amino acid sequence comprises two non-naturally occurring amino acids; and (b) crosslinking the peptide, and optionally purifying the peptide.

[0230] In some embodiments, this disclosure provides a method for synthesizing a conjugate comprising the modified peptide disclosed herein, the method comprising: (a) providing the modified peptide; and (b) derivatizing a resin-bound amine of the modified peptide with a conjugation portion on a resin. In some embodiments, the conjugation portion is the portion described herein.

[0231] In some embodiments, methods are provided herein for preparing peptides partially derivatized with PEG(n)-thiocholesterol, PEG(n)-cholesterol, thiocholesterol-PEG(n), or cholesterol-PEG(n). The fully resin-on-resin synthesis method involves (a) providing a peptide comprising at least three consecutive amino acids and two non-naturally occurring amino acids selected from sequences or variants of those in Table 1, (b) crosslinking the peptide, for example by a ruthenium-catalyzed metathesis reaction, and (c) derivatizing the N-terminus and / or C-terminus on the resin with a variable-length PEG linker connected to thiocholesterol or the cholesterol portion.

[0232] In some embodiments, the modified peptides disclosed herein are synthesized using Fmoc-based solid-phase peptide synthesis. To achieve different pinning lengths, α-methyl, α-alkenyl amino acids are mounted in specific pairings at discrete positions, such as for i, i+7 positioning, using an S-pentenylalanine residue (S5) and an R-octenylalanine residue (R8). For the pinning reaction, a Grubbs first-generation ruthenium catalyst dissolved in dichloroethane is added to the resin-bound peptide. One or more rounds of pinning can be performed, for example, three to five rounds, to ensure complete pinning. After supplementing with PEG(n)-mercaptocholesterol, PEG(n)-cholesterol, mercaptocholesterol-PEG(n), or cholesterol-PEG(n) moieties, the peptides are cleaved from the resin using trifluoroacetic acid, precipitated using a hexane:ether (1:1) mixture, air-dried, and purified by LC-MS.

[0233] In some embodiments, the peptides disclosed herein are prepared by chemical synthesis methods well known to those skilled in the art. Peptides can be synthesized using automated Merrifield solid-phase synthesis techniques on, for example, Applied Biosystems Peptide Synthesizers Model 430A or 431, using side-chain-protected amino acids and α-NH2 chemically protected with t-Boc or Fmoc.

[0234] In some embodiments, the peptides disclosed herein are prepared via solid-phase peptide synthesis (SPPS). C-terminal amino acids are attached to a cross-linked polystyrene resin via acid-labile bonds with linker molecules. This resin is insoluble in the solvents used for synthesis, allowing for relatively simple and rapid washing away of excess reagents and byproducts. The N-terminus is protected by an Fmoc group (which is stable in acid but can be removed by bases). Any side-chain functional groups are protected with base-stable, acid-labile groups.

[0235] Peptides (e.g., the peptides described herein) can be prepared in a high-throughput, combinatorial manner, for example using high-throughput multichannel combinatorial synthesizers available from, for example, Advanced Chemtech or GyrosProtein Technologies. Peptide bonds can be replaced, for example, to increase the physiological stability of the peptide: anti-anti bonds (C(O)-NH); reductamide bonds (NH-CH2); thiomethylene bonds (S-CH2 or CH2-S); oxomethylene bonds (O-CH2 or CH2-O); ethylene bonds (CH2-CH2); thioamide bonds (C(S)-NH); trans-olefin bonds (CH=CH); fluorinated trans-olefin bonds (CF=CH); ketomethylene bonds (C(O)-CHR) or CHR-C(O), where R is H or CH3; and fluorinated ketomethylene bonds (C(O)-CFR or CFR-C(O)), where R is H or F or CH3.

[0236] The peptides disclosed herein can be further modified in the following ways: acetylation, amidation, biotinylation, cinnamonylation, farnesylation, fluoresceinization, formyroxylation, myristylation, palmitoylation, and other esterifications, specifically including thiocholesterol or cholesterol modification using the resin-on-resin methods disclosed herein, phosphorylation (Ser, Tyr, or Thr), stearylation, succinylation, and thioacylation.

[0237] In some embodiments, the molecule is substantially free of unpaired peptide contaminants or is isolated. Methods for purifying the peptide include, for example, synthesizing the peptide on a solid support. Various alternative solvents and purification schemes for the separation and purification of peptides and paired peptides after cyclization are known in the art, and solvents including, but not limited to, DMSO, DMSO / dichloromethane mixtures, DMSO / NMP mixtures, or mixtures / solutions excluding DMSO can be used. DMSO / dichloromethane or DMSO / NMP mixtures may contain about 30%, 40%, 50%, or 60% DMSO. In specific cases, a 50% / 50% DMSO / NMP solution is used. The solution can be incubated for periods of 1, 6, 12, or 24 hours, followed by washing the resin, for example, with dichloromethane or NMP. In some embodiments, the resin is washed with NMP. Shaking and bubbling of an inert gas into the solution may be performed.

[0238] Determination of characterization of stabilized peptide conjugates Determination of α-helicality: The compound is dissolved in an aqueous solution ( For example 5 µM potassium phosphate solution at pH 7, or distilled H₂O to a concentration of 25 to 50 µM. Circular dichroism (CD) spectroscopy is performed using a spectrophotometer (…). For example Using standard measurement parameters on Jasco J-710, Aviv ( For example Temperature, 20 °C; wavelength, 190–260 nm; step resolution, 0.5 nm; speed, 20 nm / sec; cumulative, 10; response, 1 sec; bandwidth, 1 nm; path length, 0.1 cm. The α-helix content of each peptide was calculated by dividing the average residue ellipticity by the reported value of the model helical decapeptide (Yang et al.). Methods Enzymol. , 1986).

[0239] Determination of melting temperature (Tm): The cross-linked or unmodified template peptide is dissolved in distilled H2O or other buffer or solvent. For example The final concentration was 50 µM, and the concentration was measured using a spectropolarimeter ( For example Using standard parameters on Jasco J-710, Aviv, within a certain temperature range ( For example Tm is determined by measuring the change in ellipticity within the range of 4℃ to 95℃, using this standard parameter. For example Wavelength: 222 nm; Step resolution: 0.5 nm; Speed: 20 nm / sec; Cumulative: 10; Response: 1 sec; Bandwidth: 1 nm; Temperature increase rate: 1 °C / min; Path length: 0.1 cm.

[0240] In vitro protease resistance assay: The amide bonds of the peptide backbone are readily hydrolyzed by proteases, thus facilitating rapid degradation of the peptide compound in vivo. However, peptide helix formation typically buries and / or twists and / or shields the amide backbone, and therefore can prevent or significantly delay proteolytic cleavage. The peptides of this invention can be subjected to in vitro enzymatic proteolysis (…). For example Trypsin, chymotrypsin, and pepsin were used to assess any changes in degradation rate compared to the corresponding uncrosslinked or alternatively pinned peptides. For example, peptide-mimicking macrocycles and corresponding uncrosslinked peptides were incubated with trypsin agarose, and the reaction was quenched at different time points by centrifugation and subsequent HPLC injection to quantify residual substrate by UV absorption at 280 nm. Briefly, peptide-mimicking macrocycles and peptide-mimicking precursors (5 mcg) were incubated with trypsin agarose (Pierce) (S / E approx. 125) for 0, 10, 20, 90, and 180 min. The reaction was quenched by benchtop centrifugation at high speed; residual substrate in the separated supernatant was quantified by HPLC-based peak detection at 280 nm. The proteolytic reaction exhibited first-order kinetics, and the rate constant k was determined by a plot of ln[S] versus time.

[0241] Peptide mimics macrocycles and / or corresponding uncrosslinked peptides can each be reacted with fresh mouse, rat, and / or human serum ( example likeIncubate together (1 to 2 mL) at 37°C For example 0, 1, 2, 4, 8, and 24 hours. Samples with different macrocyclic concentrations can be prepared by serial dilution with serum. To determine the level of intact compounds, the following procedure can be used: Extract the sample, for example, by transferring 100 µL of serum to a 2 ml centrifuge tube, followed by adding 10 µL of 50% formic acid and 500 µL of acetonitrile, and centrifuging at 14,000 RPM for 10 min at 4 °C ± 2 °C. Then transfer the supernatant to a fresh 2 ml tube and evaporate on a Turbovap at 37 °C with N2 < 10 psi. Reconstitute the sample in 100 µL of 50:50 acetonitrile:water and submit for LC-MS / MS analysis. Equivalent or similar procedures for testing in vitro stability are known and can be used to determine the stability of macrocycles in serum.

[0242] Plasma stability assay ： Stable peptide stability can be tested in freshly drawn mouse plasma collected in lithium heparin tubes. Triple incubation is performed using 500 μL plasma doped with a single 10 μM peptide. The sample is gently shaken in a fixed-track shaker at 37°C, and 25 μL aliquots are removed at 0, 5, 15, 30, 60, 240, 360, and 480 min and added to 100 μL of a mixture containing 10% methanol:10% water:80% acetonitrile to prevent further peptide degradation. Samples are allowed to be placed on ice during assays and then transferred to a MultiScreen Solvinert 0.45 μm low-binding hydrophilic PTFE plate (Millipore). The filtrate is analyzed directly by LC-MS / MS. The peptide is detected as a doubly or tricharged ion using a Sciex 5500 mass spectrometer. The percentage of remaining peptide is determined by the reduction in peak area, and a logarithmic transformation is performed to calculate the half-life.

[0243] In vivo protease resistance assay ： Analytical assays based on liquid chromatography / mass spectrometry were used to detect and quantify the levels of chelated peptides in plasma. For pharmacokinetic analysis, the peptides were dissolved in sterile aqueous 5% dextran (1 mg / mL) and administered via tail vein bolus or intraperitoneal injection. For exampleC57BL / 6 mice (Jackson Laboratory) were administered 5, 10, 25, and 50 mg / kg of aristotelian peptides. Blood was collected via retroorbital puncture at 5, 30, 60, 120, and 240 minutes after administration to five animals at each time point. Plasma was harvested after centrifugation (2,500 xg, 5 min, 4°C) and stored at -70°C until assay. The concentration of aristotelian peptides in plasma was determined by reversed-phase high-performance liquid chromatography combined with electrospray ionization mass spectrometry. wait people , Journal of Proteome Research ., 2007; Walden et al. , Analytical and Bioanalytical Chem ., 2004). The study sample was compared with a series of seven peptide calibration standards in plasma at concentrations ranging from 1.0 to 50.0 µg / mL, drug-free plasma with or without internal standards, and three quality control samples ( ). For example The concentrations (3.75, 15.0, and 45.0 µg / mL) were determined together. A standard curve was constructed by plotting the analyte / internal standard peak area ratio against the known drug concentration in each calibration standard. Linear least squares regression was performed, with weights proportional to the reciprocal of the analyte concentration normalized relative to the number of calibration standards. The drug concentration in the study samples was calculated using the slope and y-intercept of the best-fit line. Plasma concentration-time curves were analyzed using a standard non-compartmental method with WinNonlin Professional 5.0 software (Pharsight Corp., Cary, NATO) to obtain pharmacokinetic parameters such as initial and terminal plasma half-lives, peak plasma levels, total plasma clearance, and apparent volume of distribution.

[0244] In vitro binding assays: To assess the binding and affinity of peptide-mimicking macrocycles and peptide-mimicking precursors to receptor proteins, fluorescence polarization assays (FPA) can be used, for example. FPA techniques use polarized light and fluorescent tracers to measure molecular orientation and migration. When excited with polarized light, molecules or peptides attach to and then bind to proteins with high apparent molecular weights (…). For example Fluorescent tracers that bind to FITC-labeled peptides that bind to large proteins ( For example FITC), due to its association with fluorescent tracers that are individually attached to smaller molecules or peptides ( For example, Compared to FITC-labeled peptides free in solution, they have a slower rotation rate when bound to proteins, and therefore emit a higher level of polarized fluorescence.

[0245] In vitro displacement assays to characterize peptide-protein interactions: To assess the binding and affinity of compounds that antagonize the interaction between antagonistic peptides and receptor proteins, fluorescence polarization assays (FPA) are used, for example, utilizing fluoresceinized peptides or peptide-mimicking macrocycles derived from template peptide sequences. FPA techniques use polarized light and fluorescent tracers to measure molecular orientation and migration. When excited with polarized light, the antagonistic peptide attaches to the molecule and then interacts with the high apparent molecular weight protein (…). For example Fluorescent tracers that bind to FITC-labeled peptides that bind to large proteins ( For example FITC), due to molecules derived solely from FITC ( For example, Compared to FITC-labeled peptides free in solution, these peptides exhibit slower rotational rates and therefore emit higher levels of polarized fluorescence. In competitive binding FPA assays, compounds antagonizing the interaction between fluoresceinized peptides and receptor proteins, such as unlabeled pinned peptides and their conjugates, can be detected, and the differential potency of compounds in disrupting the interaction can be quantified and compared.

[0246] Cell localization assay: To measure the localization of peptides or cross-linked polypeptides on or within cells, intact cells were incubated with fluorescein cross-linked polypeptides derived from PEG(n)-mercaptocholesterol or PEG(n)-cholesterol (5 µM) in serum-free or serum-supplemented medium at 37°C for 4 hours. Cells were washed twice with medium and incubated with trypsin (0.25%) at 37°C for 10 min. Cells were washed again and resuspended in PBS. For example, this was achieved using a FACSCalibur flow cytometer or Cellomics' KineticScan. RTM HCS reader is used to analyze cell fluorescence.

[0247] Example The following examples are provided to better illustrate the claimed invention, and these examples should not be construed as limiting the scope of the invention. The specific materials mentioned are for illustrative purposes only and are not intended to limit the invention. Those skilled in the art can develop equivalent means or reactants without exercising inventive capacity or departing from the scope of the invention.

[0248] Example 1. Design of anti-influenza peptides The following examples illustrate the process of designing therapeutic peptides believed to prevent conformational changes in influenza virus hemagglutinin (HA) proteins and inhibit the fusion of influenza virus particles with the host cell membrane. These therapeutic peptides are designed for several different influenza subtypes and can therefore serve as pan-influenza treatment and / or prevention.

[0249] A series of conformational changes in the influenza virus HA protein mediate the fusion of the viral particle with the host cell, triggered by a decrease in pH in the host cell's endosomal environment from neutral to acidic conditions. In the crucial final conformational change, the C-terminal helix of HA2 (residues 106 to 125 of HA2) folds back into the HR2 region (residues 77 to 101 of HA2), bringing the viral membrane and the host endosomal membrane into close proximity, while simultaneously cleaving the HA1 domain (Benton et al., ...). Nature July 2020; 583(7814):150-153. Similar to the HR1 / HR2 interaction in other viruses, the C-terminal helix / HR2 interaction supports the post-fusion structure required for influenza infection. Therefore, inhibiting the foldback of the HA2 C-terminal helix is ​​a possible target for influenza prevention.

[0250] Based on the final conformational change of the influenza HA, a therapeutic peptide mimicking the structure of the influenza HA2 C-terminal helix may interact with the HR2 domain and block the foldback of the native influenza HA2 C-terminal helix, thus preventing viral particle fusion and infection. For the initial design of the therapeutic peptide, a portion of the H3N2 HA2 C-terminal helix (DLTDSEMNKLFEKTRRQLRENA, SEQ ID NO: 5) was selected as the basis for design. To this end, the conservation of the amino acid sequence of SEQ ID NO: 5 among influenza subtypes was analyzed to determine whether this peptide sequence could be applied to more than one influenza subtype. Table 7 shows the homology of the HA region corresponding to the amino acids of SEQ ID NO: 5 among subtypes H3N2, H1N1, and influenza B.

[0251] Table 7. Conservation of C-terminal helical residues

[0252] * Residues involved in the interaction between the C-terminal helix of HA2 and the HR2 region of HA.

[0253] The data in Table 7 show that the amino acids in the C-terminal helix of H3N2 HA2, including those in contact with the HR2 region, are highly conserved between H1N1 and H3N2, but exhibit more variation in influenza B.

[0254] To further analyze the conservation of SEQ ID NO: 5 across influenza virus subtypes, the NCBI Influenza Virus Resource (NIVR) database was reviewed. The NIVR database contains 77,280 influenza A sequences and 26,225 influenza B sequences isolated from human hosts. This database shows that H1 and H3 HA proteins account for 92.5% of identified human influenza A infections. Analysis of sequences from influenza A subtypes listed in the database with at least 50 sequences indicated that the amino acids involved in C-terminal helical interactions are highly conserved. Figure 1 In influenza A subtypes, the residues that interact with the peptide based on the C-terminal helix of H3N2 HA2 are the same, except for residues 105, 108, and 140, which are highly conserved residues, namely E and Q, L and I, and F and I, respectively.

[0255] Based on this sequence analysis, therapeutic peptides were designed using amino acid sequences similar to all or part of SEQ ID NO: 5, wherein two amino acids were replaced by non-naturally occurring amino acids to crosslink or staple the peptide and stabilize its helical structure. For example, based on mapping the internal contacts within the C-terminal helix and their interaction with the HA2 domain, the hydrogen bond between E120 and R127 appears to be a plausible staplement site within the helical region. Figure 2 To generate the stabilizing peptide, Glu12 in SEQ ID NO: 5 was replaced with R8 (R-octenylalanine) and Arg19 in SEQ ID NO: 5 was replaced with S5 (S-pentenylalanine).

[0256] Example 2. Preparation of conjugated anti-influenza peptide conjugates Based on the overall design of the anti-influenza peptide described in Example 1, several exemplary peptides were prepared. These peptides were prepared by combining two naturally occurring amino acids... i, i+7 Location( Right now The amino acids at the 7-amino acid side junction were replaced with non-natural R-octenyl alanine (R8) and S-2-(4′-pentenyl) alanine (S5) to produce a pinning across two α-helical loops, or... i, i+4 Two S5 non-natural amino acids were substituted at the position to generate a pinning conjugate spanning one α-helix loop, and pinned peptide conjugates carrying C-terminal derivatizations of either PEG(n)-thiocholesterol or PEG(n)-cholesterol moieties were designed. The asymmetric synthesis of the α,α-disubstituted amino acids was carried out according to the previously detailed method (Schafmeister et al.). J. Am. Chem. Soc. , 2000; Walensky et al., Science , 2004; Bird et al. Current Protocols in Chemical Biology , 2011).

[0257] In short, the fully resin-bound peptide is capped with an acetyl group (using acetic anhydride), followed by deprotection of the C-terminal side-chain lysine amine by treatment with 2% hydrazine. The amine is acylated with Fmoc-protected PEG(n) amino acids (e.g., n = 1 to 36; e.g., 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, or 36), at which point the olefins of non-natural amino acids (e.g., R8 and S5) are crosslinked by treatment with a Grubbs(I) catalyst. The Fmoc is removed from the C-terminal NH of the PEG(n) amino acid, and the amine is acylated with carboxyl-cholesterol (prepared as described below). The final peptide product was obtained after deprotection and cleavage of the peptide, followed by purification by reversed-phase high-performance liquid chromatography / mass spectrometry (LC / MS). See also Figure 3 The complete synthesis scheme is described in the text.

[0258] Carboxy-cholesterol was prepared according to the following scheme:

[0259] Magnesium shavings were dried in an oven before the reaction. Ultrapure argon and carbon dioxide were used. Magnesium shavings (1.95 g, 0.080 mol) in anhydrous THF (450 mL) were added to an oven-dried 3-necked flask equipped with a suitable diaphragm, reflux condenser, and dropping funnel under an atmosphere of dry argon. Ethyl bromide (1.81 g, 0.016 mol) was added dropwise (20 min) via syringe, allowing the mixture to stir while slowly heating. Cholesterol chloride (25 g, 0.061 mol) in anhydrous THF (350 mL) was then added dropwise via dropping funnel, maintaining a temperature of gentle reflux. The mixture was then allowed to stir under gentle reflux. The reaction mixture was then cooled to room temperature and gently bubbled with dry carbon dioxide. The mixture was then poured into an ice-cold solution (500 mL) of 10% sulfuric acid. The mixture was then extracted with EtOAc (750 mL x 3), and the combined EtOAc extracts were dried over sodium sulfate, filtered, and the EtOAc was distilled off under reduced pressure. The resulting crude residue was ground with benzene (150 mL x 2) and collected by filtration to give a white solid (7.1 g). The solid was air-dried under vacuum.

[0260] Example 3. In vitro evaluation of stabilizing peptides against influenza A (H3N2) in MDCK cells. The in vitro antiviral activity of compounds (e.g., the modified peptides described herein) against influenza A / Perth / 16 / 2009 (H3N2) was tested in MDCK cells. The test medium was basic essential medium (MEM) supplemented with 10 U / mL trypsin, 1 µg / mL EDTA, and 50 µg / mL gentamicin. Cell plates were prepared one day prior to treatment and infection, and cells were seeded at 3e4 cells / well in 96-well plates and incubated overnight at 37 ± 2 °C, 5% CO2.

[0261] The compound was received in a 10 mM solution. The compound was serially diluted in test medium using eight 2-fold dilutions to achieve an initial (high) test concentration of 10 µM. Each dilution was added to five wells of a 96-well plate containing cells. Three wells of each concentration were infected with the virus, and two wells remained uninfected as a virulence control. Six wells were infected and untreated as a virus control, and six wells were uninfected and untreated as a cell control. The virus was prepared at an MOI of 0.001. Ribavirin was tested in parallel as a positive control. The plate was incubated at 37 °C ± 2 °C in 5% CO2.

[0262] On day 4 post-infection, once the untreated virus control wells reached maximum CPE, the plate was stained with neutral red dye for approximately 2 hours (±15 minutes). The dye in the supernatant was removed and the wells were rinsed with PBS. The incorporated dye was extracted in 50:50 Sorensen citrate buffer / ethanol for >30 minutes, and the optical density was read at 540 nm on a spectrophotometer. The optical density was converted to a percentage of the cell control and normalized relative to the virus control, and then the concentration of the test compound required to inhibit CPE by 50% (EC50) was calculated by regression analysis. Similarly, the concentration of the compound that caused 50% cell death in the absence of virus (CC50) was calculated. The selection index (SI) was calculated by dividing CC50 by EC50.

[0263] The results of this analysis are summarized in Table 8.

[0264] Table 8.

[0265] R1 has [Xaa] w = Modified peptide of SEB*NKLF (SEQ ID NO: 128); [Xaa] x = KTRRQL(SEQ ID NO: 55);[Xaa] y = E; and N-terminal conjugates having the following structures: -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] 10 -N(R5 )C(O)-(CH2)-O-cholesterol))-C(O)-NH2, where B* is leucine.

[0266] Example 4. In vitro evaluation of pinning peptides against influenza A (H3N2) in MA-104 cells. The in vitro antiviral activity of compounds (e.g., the modified peptides described herein) against influenza A / Perth / 16 / 2009 (H3N2) was tested in MA-104 cells. The test medium was MEM supplemented with 10 U / mL trypsin, 1 µg / mL EDTA, and 50 µg / mL gentamicin. Cell plates were prepared one day prior to treatment and infection, and cells were seeded at 6e4 cells per well in 96-well plates and incubated overnight at 37 ± 2 °C, 5% CO2.

[0267] The compound was received in a 10 mM solution. The compound was serially diluted in test medium using four 2-fold dilutions to achieve an initial (high) test concentration of 5 µM. Each dilution was added to five wells of a 96-well plate containing cells. Three wells of each concentration were infected with the virus, and two wells remained uninfected as a virulence control. Six wells were infected and untreated as a virus control, and six wells were uninfected and untreated as a cell control. The virus was prepared with an MOI < 0.001. Ribavirin was tested in parallel as a positive control. The plate was incubated at 37 °C ± 2 °C in 5% CO2.

[0268] On day 3 post-infection, once the untreated virus control wells reached maximum CPE, the plate was stained with neutral red dye for approximately 2 hours (±15 minutes). The dye in the supernatant was removed and the wells were rinsed with PBS. The incorporated dye was extracted in 50:50 Sorensen citrate buffer / ethanol for >30 minutes, and the optical density was read at 540 nm on a spectrophotometer. The optical density was converted to a percentage of the cell control and normalized relative to the virus control, and then the concentration of the test compound required to inhibit CPE by 50% (EC50) was calculated by regression analysis. Similarly, the concentration of the compound that caused 50% cell death in the absence of virus (CC50) was calculated. The selection index (SI) was calculated by dividing CC50 by EC50.

[0269] The results of this analysis are summarized in Table 9.

[0270] Table 9.

[0271] Example 5. In vitro evaluation of pinned peptides against influenza A and influenza B. The in vitro antiviral activity of compounds (e.g., the modified peptides described herein) against influenza A / California / 07 / 2009(H1N1)pdm09, influenza A / Perth / 16 / 2009(H3N2), and influenza B / Brisbane / 60 / 2008 (Victoria lineage) was tested in MDCK cells. The test medium was MEM supplemented with 10 U / mL trypsin, 1 µg / mL LEDTA, and 50 µg / mL gentamicin. Cell plates were prepared one day prior to treatment and infection, and cells were seeded at 6e4 cells per well in 96-well plates and incubated overnight at 37 ± 2 °C and 5% CO2.

[0272] The compound was received in a 10 mM solution. The compound was serially diluted in test medium using eight half-log10 dilutions to achieve an initial (high) test concentration of 10 µM. Each dilution was added to five wells of a 96-well plate containing cells. Three wells of each concentration were infected with the virus, and two wells remained uninfected as a virulence control. Six wells were infected and untreated as a virus control, and six wells were uninfected and untreated as a cell control. The virus was prepared with an MOI < 0.001. Ribavirin was tested in parallel as a positive control. The plate was incubated at 37 °C ± 2 °C in 5% CO2.

[0273] On days 4–5 post-infection, once the untreated virus control wells reached maximum CPE, the plate was stained with neutral red dye for approximately 2 hours (±15 minutes). The dye in the supernatant was removed and the wells were rinsed with PBS. The incorporated dye was extracted in 50:50 Sorensen citrate buffer / ethanol for >30 minutes, and the optical density was read at 540 nm on a spectrophotometer. The optical density was converted to a percentage of the cell control and normalized relative to the virus control, and then the concentration of the test compound required to inhibit CPE by 50% (EC50) was calculated by regression analysis. Similarly, the concentration of the compound that caused 50% cell death in the absence of virus (CC50) was calculated. The selection index (SI) was calculated by dividing CC50 by EC50.

[0274] The results of this analysis are summarized in Table 10.

[0275] Table 10.

[0276] While we have described several embodiments of the invention, it will be apparent that our basic examples can be modified to provide other embodiments utilizing the peptides and methods of the invention. Therefore, it should be understood that the scope of the invention should be defined by the appended claims rather than by the specific embodiments illustrated by way of example.

Claims

1. A modified peptide comprising at least 30% identical amino acid sequence to an amino acid sequence selected from the group consisting of: At least two of the amino acids contain side chains that are linked together (e.g., to form a tether).

2. A modified peptide comprising an amino acid sequence having 0 to 10 substitutions, insertions and / or deletions relative to SEQ ID NO: 1 to 6, wherein at least two amino acids comprise side chains linked together (e.g., to form a pin).

3. The modified peptide according to claim 1 or 2, wherein the side chains linked together are side chains of two amino acids separated by the following: (i) 3 amino acids (e.g., the side chains of the amino acids at positions i and i+4); or (ii) 6 amino acids (e.g., the side chains of the amino acids at positions i and i+7).

4. The modified peptide according to claim 3, wherein the side chains linked together are side chains of two amino acids separated by three amino acids (e.g., the side chains of the amino acids at positions i and i+4).

5. The modified peptide according to claim 3, wherein the side chains linked together are side chains of two amino acids separated by 6 amino acids (e.g., the side chains of the amino acids at positions i and i+7).

6. The modified peptide according to any one of the preceding claims, wherein the amino acid sequence is at least 70% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 6.

7. The modified peptide according to any one of the preceding claims, wherein the amino acid sequence is at least 80% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 6.

8. The modified peptide according to any one of the preceding claims, wherein the amino acid sequence is at least 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 6.

9. The modified peptide according to any one of the preceding claims, wherein the amino acid sequence is at least 95% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 6.

10. The modified peptide according to any one of the preceding claims, wherein the amino acid sequence is at least 98% identical to the amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 6.

11. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

1.

12. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

2.

13. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

3.

14. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

4.

15. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

5.

16. The modified peptide according to any one of claims 1 to 10, wherein the amino acid sequence is SEQ ID NO:

6.

17. The modified peptide according to any one of the preceding claims, wherein the modified peptide is conjugated with a lipid, carbohydrate, or peptide portion.

18. The modified peptide of claim 17, wherein the modified peptide is conjugated to a lipid, carbohydrate, or peptide moiety via a PEG moiety.

19. The modified peptide of claim 18, wherein the PEG portion is attached at or near the C-terminus of the amino acid sequence.

20. The modified peptide of claim 18, wherein the PEG moiety is attached at or near the N-terminus of the amino acid sequence.

21. The modified peptide according to any one of claims 17 to 20, wherein the modified peptide further comprises optionally substituted C5-C. 100 Aliphatic lipids.

22. The modified peptide of claim 21, wherein the lipid portion is a fatty acid, phospholipid, sphingolipid, or sterol.

23. The modified peptide according to claim 21 or 22, wherein the lipid portion is cholesterol, mercaptocholesterol, sitosterol, ergosterol, or tocopherol.

24. The modified peptide of claim 23, wherein the lipid portion is cholesterol.

25. A modified peptide, wherein the modified peptide has formula I': Or its pharmaceutically acceptable salt, wherein: Each Xaa independently represents a natural or non-natural amino acid; w, x, and y are independently 0 to 50, including end values; X L1 and X L2 It consists of amino acids with side chains linked together; and [Xaa] x Represents an amino acid sequence containing at least three consecutive amino acids of the following: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to the amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to the amino acid sequence selected from Table 1. [Xaa] w and [Xaa] y Each of these independently represents an amino acid sequence containing one or more consecutive amino acids: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. Choose any one of [Xaa] w The N-terminal amino acid and / or [Xaa] in y The C-terminal amino acid in it is conjugated to lipids, carbohydrates or peptides.

26. A modified peptide, wherein the modified peptide has Formula I: (I)， Or its pharmaceutically acceptable salt, wherein: R 1 and R 2 Each is independently hydrogen or optionally substituted C 1-10 Aliphatic groups; R 3 Represents the attached side chain group and is optionally substituted divalent C. 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—; Each R′ is independently hydrogen or optionally substituted C. 1-12 aliphatic; Each Xaa independently represents a natural or non-natural amino acid; w and y are each independently between 0 and 50; x is between 3 and 10; and [Xaa] x Represents an amino acid sequence containing at least three consecutive amino acids of the following: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to the amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to the amino acid sequence selected from Table 1. [Xaa] w and [Xaa] y Each of these independently represents an amino acid sequence containing one or more consecutive amino acids: Selected from the amino acid sequences in Table 1; or (i) is at least 30% identical to an amino acid sequence selected from Table 1 and / or (ii) has 1 to 10 substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from Table 1. Choose any one of [Xaa] w The N-terminal amino acid and / or [Xaa] in y The C-terminal amino acid in it is conjugated to lipids, carbohydrates or peptides.

27. The modified peptide according to claim 25 or 26, wherein [Xaa] w [Xaa] x and [Xaa] y Each of these represents an amino acid sequence containing at least three consecutive amino acids: Selected from the amino acid sequences of SEQ ID NO: 1 to 6; or (i) is at least 90% identical to an amino acid sequence selected from SEQ ID NO: 1 to 6 and / or (ii) has 1 to 10 substituted, inserted, and / or deleted amino acid sequences relative to an amino acid sequence selected from SEQ ID NO: 1 to 6. Choose any one of [Xaa] w The N-terminal amino acid and / or [Xaa] in y The C-terminal amino acid in it is conjugated to lipids, carbohydrates or peptides.

28. The modified peptide according to any one of claims 25 to 27, wherein [Xaa] w [Xaa] x and [Xaa] y Each of them represents an amino acid sequence containing at least three consecutive amino acids selected from the amino acid sequences of SEQ ID NO: 1 to 6.

29. The modified peptide according to any one of claims 25 to 27, wherein [Xaa] w [Xaa] x and [Xaa] y Each of the following represents an amino acid sequence comprising at least three consecutive amino acids: (i) being at least 90% identical to an amino acid sequence selected from SEQ ID NO: 1 to 6 and / or (ii) having one to ten substituted, inserted and / or deleted amino acid sequences relative to an amino acid sequence selected from SEQ ID NO: 1 to 6.

30. The modified peptide according to any one of claims 25 to 29, wherein [Xaa] y The C-terminal amino acid in it is conjugated to lipids, carbohydrates or peptides.

31. The modified peptide according to any one of claims 25 to 30, wherein [Xaa] w The N-terminal amino acid in it is conjugated to lipids, carbohydrates or peptides.

32. The modified peptide according to any one of claims 25 to 31, wherein w is 7.

33. The modified peptide according to any one of claims 25 to 32, wherein x is 6.

34. The modified peptide according to any one of claims 25 to 33, wherein y is 0 to 5.

35. The modified peptide according to any one of claims 25 to 34, wherein [Xaa] w Selected from Table 2.

36. The modified peptide according to any one of claims 25 to 35, wherein [Xaa] x Selected from Table 3.

37. The modified peptide according to any one of claims 25 to 35, wherein [Xaa] y Selected from Table 4.

38. The modified peptide according to any one of claims 25 to 37, wherein [Xaa] w [Xaa] x and [Xaa] y Selected from Table 5.

39. The modified peptide according to any one of claims 1 to 38, wherein the length of the modified peptide is at most 50 amino acids (i.e., w+x+y+2 ≤ 50).

40. The modified peptide according to any one of claims 1 to 39, wherein the length of the modified peptide is at most 30 amino acids (i.e., w+x+y+2 ≤ 30).

41. The modified peptide according to any one of claims 26 to 40, wherein R 1 and R 2 Each is independently hydrogen or optionally halogenated, hydroxylated, or C-substituted. 1-3 alkoxy-substituted C 1-10 Aliphatic.

42. The modified peptide according to any one of claims 26 to 41, wherein R 1 C 1-10 alkyl.

43. The modified peptide according to any one of claims 26 to 42, wherein R 1 It is a methyl group.

44. The modified peptide according to any one of claims 26 to 43, wherein R 2 C 1-10 alkyl.

45. The modified peptide according to any one of claims 26 to 44, wherein R 2 It is a methyl group.

46. ​​The modified peptide according to any one of claims 26 to 45, wherein the modified peptide has the formula IB: IB Or its pharmaceutically acceptable salt.

47. The modified peptide according to any one of claims 26 to 45, wherein the modified peptide is selected from: (I-B1), (I-B2), or (I-B3), Or its pharmaceutically acceptable salt.

48. The modified peptide according to any one of claims 26 to 47, wherein R 3 For optional substituted divalent C containing at least one double bond 3-20 A hydrocarbon chain, wherein one or more methylene units of the hydrocarbon chain are optionally and independently substituted with —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2—, —S(O)2N(R′)—, —C(O)S— or —C(O)O—.

49. The modified peptide according to any one of claims 26 to 48, wherein R 3 For optional substituted divalent C 3-20 Hydrocarbon chain.

50. The modified peptide according to any one of claims 26 to 49, wherein R 3 C 7-12 Alkenyl group.

51. The modified peptide according to any one of claims 26 to 50, wherein R 3 It is a C8 subene group.

52. The modified peptide according to any one of claims 26 to 50, wherein R 3 C 11 Alkenyl group.

53. The modified peptide according to any one of claims 26 to 48, wherein R 3 -(CH2) 1-7 -CH=CH-(CH2) 1-7 - 54. The modified peptide according to any one of claims 26 to 48, wherein R 3 -(CH2) 3-7 -CH=CH-(CH2) 3-4 - 55. The modified peptide according to any one of claims 26 to 48, wherein R 3 It is -(CH2)6-CH=CH-(CH2)3-.

56. The modified peptide according to any one of claims 26 to 48, wherein R 3 It is -(CH2)3-CH=CH-(CH2)3-.

57. The modified peptide according to any one of claims 26 to 56, wherein the modified peptide has formula II': II' Or its pharmaceutically acceptable salt, wherein: R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain, wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -O-, -N(R′)-, -S-, -OC(O)-, -C(O)O-, -C(O)-, -S(O)-, -S(O)2-, -N(R′)S(O)2-, -S(O)2N(R′)-, -N(R′)C(O)-, -C(O)N(R′)-, -OC(O)N(R′)-, –N(R′)C(O)O- or -[O-CH2CH2] n Replace, where n is from 1 to 20; and R 6 For example, conjugated parts (e.g., lipids, carbohydrates, or peptides).

58. The modified peptide according to claim 57, wherein R 4 C is a divalent linear or branched, saturated or unsaturated, optionally substituted C 1-30 A hydrocarbon chain, wherein 1 to 10 methylene units of the hydrocarbon chain are independently derived from -N(R′)C(O)-, -C(O)N(R′)-, or -[O-CH2CH2]. n Replacement, where n is from 1 to 20.

59. The modified peptide according to claim 57 or 58, wherein R 6 It is the lipid portion.

60. The modified peptide according to any one of claims 57 to 59, wherein: R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-6 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-6 (alkylene)-R 6 ))-C(O)-NH2; R 5 It is hydrogen or C 1-4 alkyl; R 6 C5-C is optionally replaced 100 Aliphatic lipid fraction; and n can be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

61. The modified peptide according to any one of claims 57 to 60, wherein R 6 Selected from: , , , ,or Each of these can be substituted at will.

62. The modified peptide according to claim 60 or 61, wherein R 6 Optionally halogenated, C 1-3 Alkyl, hydroxyl or C 1-3 Alkyl-substituted.

63. The modified peptide according to any one of claims 57 to 62, wherein the modified peptide has formula IIa1: IIa1 Or its pharmaceutically acceptable salt, wherein: R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-6 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-6 (alkylene)-R 6 ))-C(O)-NH2; R 5 It is hydrogen or C 1-4 alkyl; R 6 One of the following: , , , or Each of them may be halogenated or C-substituted once or more. 1-3 Alkyl, hydroxyl or C 1-3 Alkyl substitution; n is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16; p is 2, 3, 4, 5, 6, 7, or 8; and q can be 2, 3, 4, 5 or 6.

64. The modified peptide according to claim 63, wherein the modified peptide has formula IIa: IIa Or its pharmaceutically acceptable salt.

65. The modified peptide according to any one of claims 57 to 64, wherein R 4 -N(H)-C(H)((CH2)4N(H)(C(O)-(C 2-3 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-2 (alkylene)-R 6 ))-C(O)-NH2.

66. The modified peptide according to any one of claims 57 to 64, wherein R 4 is -N(H)-C(H)((CH2)4N(H)(C(O)-CH2CH2-[O-CH2CH2] n -N(R 5 )C(O)-R 6 ))-C(O)-NH2.

67. The modified peptide according to any one of claims 57 to 64, wherein -R 4 -R 6 for 。 68. The modified peptide according to any one of claims 57 to 64, wherein -R 4 -R 6 for 。 69. The modified peptide according to any one of claims 57 to 62, wherein the modified peptide has formula IIb: IIb Or a pharmaceutically acceptable salt thereof, wherein R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] n -N(R 5 )C(O)-R 6 .

70. The modified peptide according to claim 69, wherein the modified peptide has formula IIc: IIc Or a pharmaceutically acceptable salt thereof, wherein R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] n -N(R 5 )C(O)-R 6 .

71. The modified peptide according to any one of claims 57 to 70, wherein the modified peptide has formula V: V in R 9 -C(O)-(C 2-6 (alkylene)-[O-CH2CH2) n -N(R 5 )C(O)-(C 0-6 (alkylene)-R 6 ; R 5 It is hydrogen or C 1-4 alkyl; R 6 for It is optionally halogenated, C 1-3 Alkyl, hydroxyl or C 1-3 Alkyl substitution; R 8 -C(O)-(C 1-4 Alkyl); and n can be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

72. The modified peptide according to any one of claims 57 to 71, wherein R 6 for It undergoes one or more halogenation, C 1-3 Alkyl, hydroxyl or C 1-3 Alkyl-substituted.

73. The modified peptide according to any one of claims 57 to 71, wherein R 6 for .

74. The modified peptide according to any one of claims 57 to 71, wherein R 6 for .

75. The modified peptide according to any one of claims 71 to 74, wherein R 8 It is -C(O)CH3.

76. The modified peptide according to any one of claims 60 to 75, wherein R 5 It is hydrogen.

77. The modified peptide according to any one of claims 57 to 76, wherein n is 4.

78. The modified peptide according to any one of claims 57 to 76, wherein n is 8.

79. The modified peptide according to any one of claims 57 to 76, wherein n is 12.

80. The modified peptide according to any one of claims 71 to 79, wherein the modified peptide has formula VI: VI Or its pharmaceutically acceptable salt, wherein: R 9 The expression is -C(O)-(CH2CH2)-[O-CH2CH2] 12 -N(H)C(O)-R 6 ;and R 6 for .

81. A modified peptide selected from Table 6, or a pharmaceutically acceptable salt thereof.

82. The modified peptide according to any one of the preceding claims, wherein the peptide is structurally stable.

83. The modified peptide according to any one of the preceding claims, wherein the two amino acid side chains are linked together to form a staple, the staple spanning one or two turns of the α-helix of the modified peptide.

84. A pharmaceutical composition comprising a modified peptide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 83, and a pharmaceutically acceptable carrier.

85. A method for treating or preventing influenza virus infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a modified peptide or a pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 83, or a pharmaceutical composition as described in claim 84.

86. A method for inhibiting the fusion of influenza virus particles with host cells in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a modified peptide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 83, or a pharmaceutical composition according to claim 84.

87. A method for inhibiting conformational changes in influenza hemagglutinin (HA) protein in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a modified peptide or a pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 83, or a pharmaceutical composition as described in claim 84.

88. The method according to any one of claims 85 to 87, wherein the influenza is an influenza A virus.

89. The method according to claim 88, wherein the influenza A virus is subtype H10N4, subtype H10N5, subtype H10N7, subtype H10N8, subtype H10N9, subtype H11N1, subtype H11N13, subtype H11N2, subtype H11N4, subtype H11N6, subtype H11N8, subtype H11N9, subtype H12N1, subtype H12N4, subtype H12N5, subtype H12N8, subtype H13N2, or subtype H... 13N3, subtype H13N6, subtype H13N7, subtype H14N5, subtype H14N6, subtype H15N8, subtype H15N9, subtype H16N3, subtype H1N1, subtype H1N2, subtype H1N3, subtype H1N6, subtype H1N9, subtype H2N1, subtype H2N2, subtype H2N3, subtype H2N5, subtype H2N7, subtype H2N8, subtype H2N9, subtype H3N1, subtype H3N2, subtype H3N3, subtype H 3N4, subtype H3N5, subtype H3N6, subtype H3N8, subtype H3N9, subtype H4N1, subtype H4N2, subtype H4N3, subtype H4N4, subtype H4N5, subtype H4N6, subtype H4N8, subtype H4N9, subtype H5N1, subtype H5N2, subtype H5N3, subtype H5N4, subtype H5N6, subtype H5N7, subtype H5N8, subtype H5N9, subtype H6N1, subtype H6N2, subtype H6N3, subtype H 6N4, subtype H6N5, subtype H6N6, subtype H6N7, subtype H6N8, subtype H6N9, subtype H7N1, subtype H7N2, subtype H7N3, subtype H7N4, subtype H7N5, subtype H7N7, subtype H7N8, subtype H7N9, subtype H8N4, subtype H8N5, subtype H9N1, subtype H9N2, subtype H9N3, subtype H9N5, subtype H9N6, subtype H9N7, subtype H9N8 or subtype H9N9 or combinations thereof.

90. The method according to any one of claims 85 to 87, wherein the influenza is influenza B virus.

91. The method according to any one of claims 85, 88 or 89, wherein the modified peptide or a pharmaceutically acceptable salt thereof treats or prevents influenza virus infection caused by more than one (e.g., two, three, four, five, six, seven, eight, nine, ten or more) subtypes of influenza A virus.

92. The method according to claim 85 or 91, wherein the modified peptide or a pharmaceutically acceptable salt thereof treats or prevents influenza virus infection caused by both influenza A virus and influenza B virus.

93. The method according to any one of claims 86, 88 or 89, wherein the modified peptide or its pharmaceutically acceptable salt inhibits the fusion of more than one (e.g., two, three, four, five, six, seven, eight, nine, ten or more) influenza A virus subtypes with the host cell.

94. The method according to claim 86 or 93, wherein the modified peptide or a pharmaceutically acceptable salt thereof inhibits the fusion of both influenza A virus and influenza B virus with the host cell.

95. The method according to any one of claims 87, 88 or 89, wherein the modified peptide or its pharmaceutically acceptable salt inhibits conformational changes of more than one (e.g., two, three, four, five, six, seven, eight, nine, ten or more) influenza A virus subtypes of influenza hemagglutinin (HA) protein.

96. The method according to claim 87 or 95, wherein the modified peptide or a pharmaceutically acceptable salt thereof inhibits conformational changes in the influenza hemagglutinin (HA) protein of both influenza A and influenza B viruses.

97. The method according to any one of claims 91, 93 or 95, wherein the influenza A virus subtypes include H1N1 and H3N2.

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