Method for preparing peptide conjugates and products prepared thereby

Abstract

This application provides a method for synthesizing peptide conjugates by automated flow chemistry, comprising synthesizing a covalent framework connected to different modules, wherein the covalent framework comprises at least one diaminoaliphatic acid, preferably lysine, and a product produced thereby, for example, an antiviral peptide conjugate.

Description

[Background technology] 【0001】 Related applications This application claims the benefits of U.S. Provisional Patent Application No. 63 / 470,257, filed on 1 June 2023. All teachings of the said application are incorporated herein by reference. 【0002】 Respiratory infections caused by various viruses can lead to illnesses ranging from mild colds to severe pneumonia, particularly in vulnerable populations such as the elderly, infants, and immunocompromised individuals. Most current medications provide symptom relief, with few drugs altering the course of the disease. Paramyxoviruses, a family of negative-strand RNA viruses, include several respiratory pathogens such as human parainfluenza virus (HPIV), measles virus, and mumps virus, which can cause a variety of mild to severe respiratory illnesses, such as croup, pneumonia, and bronchiolitis. Furthermore, emerging paramyxoviruses, such as Nipah virus and Hendra virus, can cause severe respiratory and neurological disorders in humans. Respiratory syncytial virus (RSV) is a member of the paramyxovirus family and is a leading cause of hospitalization in infants under one year of age, particularly in the United States. While most RSV infections are self-limiting, severe cases can be life-threatening, and there are currently no specific antiviral therapies available for RSV. Several approaches are being developed to prevent and treat RSV infections. For example, the monoclonal antibody palivizumab has been used to prevent severe RSV infections in high-risk infants, and inhaled ribavirin has been used to treat severe RSV infections in immunocompromised patients. However, the effectiveness of these treatments is limited, and improved therapies are needed that can prevent and treat respiratory viral infections in a wider range of patients. 【0003】 Furthermore, respiratory viruses can mutate rapidly, leading to the emergence of new strains that can evade existing treatments and vaccines. Therefore, there is an urgent need to develop new therapeutic strategies that can keep pace with evolving respiratory viruses, including RSV and emerging zoonotic viruses such as SARS-CoV, MERS-CoV, and SARS-CoV-2. 【0004】 On the other hand, peptide conjugates offer a powerful therapeutic approach in the fight against viral infections, providing specificity, stability, and efficient delivery. Solid-phase synthesis is a well-established method for producing peptides by the sequential addition of protected amino acids. However, this technique has drawbacks, such as side reactions and aggregation, which can lead to reduced purity and prolonged synthesis times. To address the challenges posed by evolving viruses and to increase availability, there is a need for novel methods that enable the more efficient production of antiviral peptide conjugates. Such advances would not only be beneficial to the field of antiviral therapy but would also broaden the possibilities for developing peptide conjugates in a more readily available form overall. [Overview of the project] 【0005】 This invention provides a novel method for the efficient and accurate synthesis of peptide conjugates. It is based on the discovery that novel peptide conjugates can be synthesized on demand by using automated flow chemistry to conjugate peptides, preferably derived from viral envelope proteins (e.g., fusion proteins or spike proteins, preferably HRC peptides), and / or other competitive ligands to a hydrophobic moiety, such as a lipid molecule, preferably cholesterol. This method provides a programmable rapid-response platform for the on-demand synthesis of a variety of useful peptide conjugates, such as potent antiviral therapeutics. The platform includes a covalent framework designed to enable covalent linkage to different modules (essentially peptide and membrane-immobilized units) via automated flow chemistry. The covalent framework includes at least one diaminoliphatic acid. Preferably, the covalent framework includes multiple diaminoliphatic acids, optionally a linker, and / or optionally a spacer. Preferably, the diaminoliphatic acid is lysine. 【0006】 In some embodiments, the platform comprises four modules: a viral fusion peptide inhibitor, a membrane-immobilized unit (such as a lipid molecule), a receptor-targeting peptide, and a spike-binding peptide. A covalent combination having at least two modules is a fusion peptide inhibitor conjugated with a membrane-immobilized unit, resulting in an effective antiviral compound. The modules can be connected via a covalent framework consisting of at least one lysine molecule. Other modules consist of connectors comprising peptide linkers, polyethylene glycol (PEG) derivatives, amides, and esters. These connectors can be rapidly fabricated using automated flow chemistry. 【0007】 The present invention also provides compounds, compositions, and methods of using them for treating or preventing infectious diseases, including, for example, compounds identified by the platform. 【0008】 In some embodiments, the present invention provides compounds comprising one, two, three, or more HRC peptides of a viral fusion protein (also called a spike protein) conjugated to a membrane-fixed portion (e.g., a hydrophobic portion) via a covalent framework comprising at least one diaminoliphatic acid, preferably lysine, and optionally a spacer. The hydrophobic portion can be a membrane-integrating ligand such as a sterol (e.g., cholesterol, sitosterol, and campesterol), sphingolipid, glycolipid, or glycerophospholipid. The HRC sequence is preferably derived from a paramyxovirus fusion protein (e.g., an RSV fusion protein or an HPIV fusion protein) or a coronavirus spike protein. The HRC peptides of the present invention inhibit viral fusion. 【0009】 The present invention includes compositions for the delivery of the compounds of the present invention, for example, pulmonary delivery or nasal delivery. The present invention also provides a method for treating or preventing a viral infection in a subject requiring treatment or prevention of infection, for example, paramyxovirus infection and SARS-CoV-2 (COVID-19) infection, comprising administering an effective amount of the compounds of the present invention. The present invention further provides a method for producing the compositions of the present invention. 【0010】 In some embodiments, antiviral compounds useful for treating paramyxoviruses include compounds of the present invention having the formula shown in Figure 1A, Figure 1B, Figure 1C, or Figure 1D. 【0011】 In some embodiments, antiviral compounds useful for treating coronaviruses include compounds of the present invention having the formula shown in Figure 1E. 【0012】 The present invention is intended to provide an efficient production of molecular libraries. By the platforms and libraries that can be produced, by examining mutant sequences, antiviral activities against various viruses and their mutants, such as SARS-CoV2 and its variants, including but not limited to alpha, beta, gamma, delta, and omicron variants, can be improved, and it becomes possible to identify linkers, spacers, sequences, and targeting moieties. 【0013】 The foregoing and other objects, features, and advantages of the present invention will become apparent from the following more detailed description of the preferred embodiments of the present invention shown in the accompanying drawings, in which, even if the figures are different, the same reference numerals refer to the same parts. The drawings are not necessarily to scale, and instead, emphasis is placed on explaining the principles of the present invention. 【Brief Description of the Drawings】 【0014】 [Figure 1A] It is a diagram representing an exemplary compound of the present invention. [Figure 1B] It is a diagram representing an exemplary compound of the present invention. [Figure 1C] It is a diagram representing an exemplary compound of the present invention. [Figure 1D] It is a diagram representing an exemplary compound of the present invention. [Figure 1E] It is a diagram representing an exemplary compound of the present invention. [Figure 2] It is a schematic diagram of a system for performing peptide conjugate synthesis according to a set of embodiments. 【Modes for Carrying Out the Invention】 【0015】 Compound The present invention provides a programmable rapid response platform for on-demand synthesis of a wide variety of peptide conjugates that can retain potential applications in prevention, diagnosis, and treatment. Preferably, the peptide conjugate is a therapeutic peptide conjugate, such as an antiviral peptide conjugate. The platform comprises a therapeutic peptide, a membrane-fixed portion, and a polymer core (B) covalently linking the therapeutic peptide to the membrane-fixed portion. Preferably, the therapeutic peptide is a fusion peptide inhibitor. The platform optionally further comprises a spike-binding peptide. The platform optionally further comprises a target peptide, such as a receptor-binding domain (RBD)-targeted peptide. For simplicity, in some embodiments, the term “compound” is used to refer to a peptide conjugate. 【0016】 The compound of the present invention has the following general formula: (Peptide-linker) n -B-Membrane fixing part [In the formula, the peptide is a short-chain amino acid sequence, preferably a therapeutic peptide; the linker is optional; B is a polymeric core covalently linking each peptide portion to a hydrophobic portion, comprising one or more diaminoliphatic acids, preferably lysine (AKA, Lys, or K), and optionally a spacer; and n is an integer selected from 1, 2, 3, or more.] It can be characterized by the following: The term “diaminoaliphatic acid” refers to a molecule comprising a carboxylic acid and two amine functional groups, each functional group being covalently linked to the aliphatic portion. In preferred embodiments, the therapeutic peptide is a peptide inhibitor, preferably a fusion peptide inhibitor. The term “therapeutic peptide” refers to a short-chain amino acid sequence consisting of 4 to 100 (preferably 6 to 80 or 9 to 50) amino acid residues in length, designed or derived from natural proteins or peptides, including wild-type and mutant sequences, and having therapeutic effects. 【0017】 In preferred embodiments, the membrane-immobilized portion is a membrane-integrating lipid. In preferred embodiments, the compound is produced by chemically conjugating each portion or its base, such as an amino acid, in a sequential and continuous manner. Automated flow chemistry, including the use of a solid support, is preferably used to synthesize the compounds of the present invention. 【0018】 Peptide inhibitors are short-chain peptide molecules that can bind to specific proteins and inhibit their activity, typically consisting of 4 to 100 amino acid residues in length. The fusion peptide inhibitors used in this invention are preferably derived from a viral fusion protein, more preferably from the C-terminus of a viral fusion protein, and most preferably from the C-terminal heptad repeat (HRC) region of a viral fusion protein. In some embodiments, fusion peptide inhibitors derived from the HRC region of a viral fusion protein are referred to as "HRC peptides." The HRC region of a viral fusion protein is involved in the formation of a 6-helix bundle structure that drives viral fusion. HRC peptides mimic the HRC region and can bind to the N-terminal heptad repeat (HRN) region of a fusion protein to inhibit the formation of the 6-helix bundle, thereby preventing viral fusion and entry into host cells. Note that the acronym CHR is also used in the literature to refer to the C-terminal heptad repeat region. Therefore, "CHR peptide" or "CHR-peptide" is used to describe the same class of peptide inhibitors defined above, which is encompassed by "HRC peptide" as used in this application. In some embodiments, the HRC peptide is a wild-type peptide derived from the HRC region of a viral fusion protein. In other embodiments, the HRC peptide is a variant or mutant sequence peptide derived from the HRC region of a viral fusion protein, which may include naturally occurring gene mutations and / or modifications made in a laboratory setting. In some embodiments, the HRC peptide can have a length ranging from 5 to 100 amino acid residues, 6 to 80 amino acid residues, 8 to 60 amino acid residues, or 10 to 50 amino acid residues. In preferred embodiments, the HRC peptide has a length of 12 to 40 amino acid residues. In preferred embodiments, the HRC peptide has a length of 18 to 39 amino acid residues. In most preferred embodiments, the HRC peptide has a length of approximately 36 amino acid residues. 【0019】 An optional “linker” is defined as a divalent moiety or divalent group that covalently bonds to the peptide (preferably at its C-terminus or N-terminus) and also to B. 【0020】 The polymer core B is a multivalent moiety designed to enable rapid synthesis of the compounds of the present invention via automated flow chemistry, and can be covalently linked with fusion peptide inhibitors, membrane-immobilized moieties, and other optional modules to be modified to improve antiviral activity. B preferably comprises at least one lysine molecule, optionally a spacer, and optionally functional groups such as amides, esters, and ethers. In some embodiments, B comprises one, two, three, four, five, six, or more lysine molecules. 【0021】 In some embodiments, B contains one lysine molecule. In some embodiments, B contains two lysine molecules. In some embodiments, B contains three lysine molecules. In some embodiments, B contains four lysine molecules. In some embodiments, B contains five lysine molecules. In some embodiments, B contains six lysine molecules. If two or more lysine molecules are present in B, they may be covalently linked to each other via amide bonds, amino acid linkers such as GS linkers, or polymer linkers such as PEG linkers. In one preferred embodiment, B contains two lysine molecules covalently linked to each other via amide bonds. In a further preferred embodiment, B contains one lysine molecule. In yet another embodiment, B contains three lysine molecules covalently linked to each other via two amide bonds. In yet another embodiment, B contains four lysine molecules covalently linked to each other via three amide bonds. 【0022】 In some embodiments, B is formula (B1): [ka] [In the formula, [ka] m represents a covalent bond linking to a portion of the compound comprising one or more fusion peptide inhibitors, a membrane-immobilized portion, optionally one or more spike-binding peptides, and optionally one or more targeted peptides, where m is an integer that may be 0, 1, 2, 3, 4, or more. It is represented by. Preferably, [ka] The symbol represents a covalent bond to the -CO- group. Preferably, m is 0, 1, or 2. 【0023】 In some embodiments, the compound is of formula (I): [ka] [In the formula, the peptide, linker, membrane-immobilized portion, and m are as defined above, including in all embodiments and preferred embodiments.] This is represented by [the formula / code]. In some embodiments, each linker may be independent and not present. 【0024】 In some embodiments, B comprises one or more lysines, one or more spacers, and one or more additional functional groups such as amides, esters, or ethers. As used herein, the term “spacer” refers to a hydrophilic, biocompatible molecule or chemical group that is inserted between two lysines, between lysine and a fusion peptide inhibitor, between lysine and a membrane-fixed portion, between lysine and a spike-binding peptide, or between lysine and a targeted peptide to increase the distance between them. Spacers are used to avoid steric hindrance, reduce aggregation, and improve solubility and accessibility of the compound to its target. A common type of spacer used in compounds is polyethylene glycol (PEG), a hydrophilic, biocompatible polymer that can increase the solubility and stability of the compound in vivo. When a hydrophilic spacer PEG is present in B, a variety of PEG derivatives, such as, but not limited to, amine-PEG-carboxylate, amine-PEG-maleimide, amine-PEG-biotin, amine-PEG-azide, azide-PEG-carboxylate, amine-PEG-NHS ester, maleimide-PEG-NHS ester, and biotin-PEG-NHS ester, can be used in the synthesis of the compound. Amine-PEG-carboxylate is preferably synthesized via automated flow chemistry into H2N-PEG 1-40 -COOH, H2N-PEG 1-40 -CH2COOH, or H2N-PEG 1-40 It is preferable to use it to synthesize compounds such as -CH2CH2COOH. Other hydrophilic spacers, such as polyethyleneamine, polyacetal polymer, poly(1-hydroxymethylethylenehydroxymethyl-formal) (PHF), or carbohydrates may also be used. The length of the hydrophilic spacer can correspond to the full length of the protein gap to facilitate the orientation of the HRC peptide for binding to the HRN domain. B and the spacer, the linker and the spacer, or the peptide and the spacer may be linked to each other by residues in a chemical reaction (such as an automated flow chemical reaction). 【0025】 Therefore, in some embodiments, B is, for example, formula (B2): [ka] [In the formula, [ka] And m is as defined above, including all embodiments and preferred embodiments. It can be expressed as follows: p is an integer selected from 0 to 40. 【0026】 Therefore, when spacer PEG is present in B, in some embodiments, the compound is of formula (II): [ka] [In the formula, the peptide, linker, membrane-immobilized portion, and m are as defined above, including in all embodiments and preferred embodiments.] It can be represented by [this]. 【0027】 The membrane-fixed portion is a membrane-integrating lipid, such as cholesterol, sphingolipids, sphingomyelin, glycolipids, glycerophospholipids (e.g., phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine), ergosterol, 7-dihydrocholesterol, and stigmasterol. Cholesterol is preferred. Typically, B is directly or indirectly linked to the hydroxyl group of cholesterol, for example, 3-OH. The membrane-fixed portion facilitates the insertion of the compound of the present invention into the cell membrane and can inhibit viral entry. 【0028】 B can be covalently attached to a convenient location on the membrane-fixed portion. In some embodiments, the attachment is made via a hydroxyl group on the membrane-fixed portion. For example, if the membrane-fixed portion is cholesterol, B can be a -C(O)- group or a -C1~4 Cholesterol can be attached to an alkylene C(O)- group, such as a -CH2C(O)- group. 【0029】 In some embodiments, the membrane-fixed portion may be cholesterol. Examples of cholesterols include cholesterol, 3β-amino-5-cholestene, 3β-thiol-5-cholestene, 3β-carboxymethoxy-5-cholestene, cholesterol esters such as cholesterol hemysuccinate, cholesterol salts such as cholesterol bisulfate and cholesterol sulfate, ergosterol, ergosterol esters such as ergosterol hemysuccinate, ergosterol salts such as ergosterol bisulfate and ergosterol sulfate, lanosterol, lanosterol esters such as lanosterol hemysuccinate, and lanosterol salts such as lanosterol bisulfate and lanosterol sulfate. 【0030】 Examples of membrane-fixing groups that can be used in the present invention include, but are not limited to, fatty acids, or long-chain alkyl chains, or 33-cholesterylamine, or 3β-cholesterylthiol, or other cholesteryl analogs and derivatives. In the case of 3β-cholesterylamine type molecules, endosomal recycling of the conjugate is possible, and the fixation half-life on the cell surface is extended. The conjugate may be in monomeric, dimeric, trimer, or oligomeric form. The antibody-binding molecule may also be in monomeric, dimeric, trimer, or oligomer form within the conjugate. Examples of 3β-cholesterylamine, 3β-cholesterylamine-containing moieties, and their derivatives that can be used in the conjugate can be found in U.S. Patent Application No. 15 / 945,741. 【0031】 In some embodiments, the membrane-fixed portion may be a phospholipid. Phospholipids that can be used in this application include, but are not limited to, egg phosphatidylcholine (EPC), egg phosphatidylglycerol (EPG), egg phosphatidylinositol (EPI), egg phosphatidylserine (EPS), phosphatidylethanolamine (EPE), and phosphatidic acid (EPA), the corresponding phospholipids derived from soybeans, soy phosphatidylcholine (SPC), SPG, SPS, SPI, SPE, and SPA, the corresponding hydrogenated egg and soybean phospholipids (e.g., HEPC, HSPC), other phospholipids composed of ester bonds of fatty acids containing chains of 12 to 26 carbon atoms at the 2nd and 3rd positions of glycerol, and various head groups including choline, glycerol, inositol, serine, and ethanolamine at the 1st position of glycerol, as well as the corresponding phosphatidic acid. These fatty acid chains may be saturated or unsaturated, and phospholipids can be composed of fatty acids of varying chain lengths and degrees of unsaturation. In particular, the composition of this formulation may include dipalmitoylphosphatidylcholine (DPPC), a major component of naturally occurring pulmonary surfactants. Other examples include mixed phospholipids such as dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidecholine (DPPQ) and dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPQ) and distearoylphosphatidylglycerol (DSPG), dioleylphosphatidylethanolamine (DOPE), and palmitoylstearoylphosphatidylcholine (PSPC) and palmitoylstearolphosphatidylglycerol (PSPG), as well as monooleoylphosphatidylethanolamine (MOPE). 【0032】 In some embodiments, the membrane-anchoring moiety can be tocopherol. Tocopherols include tocopherol, esters of tocopherol such as tocopherol hemisuccinate, salts of tocopherol such as tocopherol hydrogen sulfate and tocopherol sulfate. 【0033】 In some embodiments, the linker can have 1, 2, 3, 4, 5 or more subunits or segments. In some embodiments, the linker includes subunits of one or more amino acids. The amino acids can be naturally occurring or synthetic. Thus, the linker can be (Gly) n+1 , (GlySerGly) n or (Gly-Pro) n where n is 1 or greater, for example, 1 to 12, 1 to 6, or 1 to 4. GlySerGly is an example of an amino acid sequence that can form a linker or part of a linker. 【0034】 In certain embodiments, the linker can include non-amino acid subunits. In some embodiments, examples of non-amino acid subunits of the linker are -(OCH2CH2) m - where m is 1 to 15, for example 2 to 10, 2 to 6 or 4. Introduction of (poly)ethylene glycol groups aids solubility in aqueous media. In some embodiments, examples of non-amino acid portions of the linker are -CH2C(O)- and -CH2C(O)NHCH2CH2(OCH2CH2)4C(O)-. 【0035】 For example, it may be advantageous to use a linker having three subunits. The first optional subunit is a flexible peptide such as -(G) m - or -(GS) mThe first subunit contains G-, where m is an integer of 1, 2, 3, 4, 5 or more, e.g., 2. The second subunit may be a residue from a chemical reaction (e.g., an automated flow chemical reaction), e.g., a peptide or a peptide bond involving the N-terminus, C-terminus, or side chain of the first subunit, an ester, or an ether. The residue may be non-cleavable, such as one formed with carbodiimide or sulfhydrylmaleimide. The third optional subunit may be a hydrophilic spacer similar to or identical to the optional spacer used in polymer core B, e.g., polyethylene glycol, polyethyleneamine, polyacetal polymer, poly(1-hydroxymethylethylenehydroxymethyl-formal) (PHF), or a carbohydrate. The hydrophilic spacer is generally a polymer and may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more monomers. Polyethylene glycol (PEG4) with four monomers is sufficient. The length of the hydrophilic spacer can correspond to the full length of the protein gap, facilitating the orientation of the HRC peptide for binding to the HRN domain. 【0036】 Referring to formula (B2), if the membrane-fixed portion contains cholesterol, in some embodiments, the compound is formula (III): [ka] [In the formula, peptide, linker, m, and p are as defined above, including in all embodiments and preferred embodiments.] It can be represented by [this]. 【0037】 The compounds of the present invention can be rapidly programmed to provide antiviral effects against evolving viruses by incorporating various fusion peptide inhibitors into the compounds, preferably via automated flow chemistry. The target viruses are preferably enveloped viruses having a viral spike protein or viral fusion protein, such as coronaviruses or paramyxoviruses. 【0038】 In some embodiments, the compounds of the present invention have anti-paramyxovirus activity. Paramyxoviruses are a family of enveloped negative-strand RNA viruses that can cause a wide range of respiratory and systemic diseases in humans and animals. This family includes well-known viruses such as measles virus, mumps virus, and respiratory syncytial virus (RSV). The paramyxovirus virion structure consists of a lipid envelope containing two major glycoproteins: a binding (HN, H, or G) protein and a fusion (F) protein. The binding protein is responsible for binding to host cell receptors, and the fusion protein is involved in the fusion of the viral membrane with the cell membrane, facilitating viral entry into host cells. The binding protein interacts with various cell receptors. For example, the HN in parainfluenza virus 5 (PIV5, formerly known as SV5) binds to sialic acid, the H in measles interacts with CD46 or CDw150 / SLAM, the G in Hendra and Nipah viruses binds to Ephrin B2, and the G in respiratory syncytial virus (RSV) binds to heparin sulfate. 【0039】 The paramyxovirus F protein is a type I viral fusion protein that plays a crucial role in viral entry by mediating membrane fusion between the viral envelope and the host cell membrane. The F protein is synthesized as an inactive precursor (F0), which must be cleaved by host cell proteases to produce the protein's active forms: a large C-terminal fragment (F1) and a small N-terminal fragment (F2). F1 contains a hydrophobic fusion peptide at its N-terminus, as well as two hydrophobic heptad repeat regions (HR1 and HR2). HR1 is directly adjacent to the fusion peptide, while HR2 is adjacent to the transmembrane (TM) domain, separated, for example, by approximately 250 residues. The F protein is highly conserved among paramyxoviruses and contains multiple functional domains, including a signal peptide, a fusion peptide, two heptad repeat domains HR1 and HR2, a transmembrane domain, and a cytoplasmic tail. 【0040】 The fusion process mediated by the F protein is triggered by the binding of the H protein to the host cell receptor. This interaction causes a conformational change in the F protein, exposing the fusion peptide, which then leads to its insertion into the host cell membrane. Subsequently, the HR1 and HR2 domains of the F protein form a 6-helix bundle structure, bringing the viral membrane and cell membrane closer together and facilitating membrane fusion. 【0041】 Due to the high degree of conservedness of the F protein among paramyxoviruses, the F protein is an attractive target for peptide inhibitors such as HRC peptides. HRC peptides can inhibit the paramyxovirus F protein by binding to the HR1 domain of the F protein, thereby interfering with the interaction between the HR1 and HR2 domains necessary for the fusion process. During the fusion process, the HR2 domain undergoes a conformational change to form a 6-helix bundle with the HR1 domain. HRC peptide inhibitors mimic the HR2 domain and are designed to competitively bind to the HR1 domain, preventing the formation of the 6-helix bundle and inhibiting the fusion process. As a result, the virus is unable to enter host cells, and the spread of infection is suppressed. 【0042】 The compounds of the present invention can preferably be modified to target a variety of paramyxoviruses by incorporating a fusion peptide inhibitor for the target paramyxovirus. For example, the compounds can be designed to target HPIV-3. 【0043】 Human parainfluenza virus type 3 (human respirovirus 3, HPIV-3) is a member of the paramyxovirus family and is a common cause of respiratory tract infections, particularly in infants and young children. HPIV-3 is transmitted through respiratory secretions and can cause a variety of symptoms, including cough, fever, runny nose, and difficulty breathing. In severe cases, it can lead to pneumonia and bronchiolitis. As of the time of this application, no vaccine is currently available for HPIV-3, and treatment mainly concerns supportive care. 【0044】 In some embodiments, the HPIV-3-targeting fusion peptide inhibitor is a short-chain peptide derived from the HRC region of the HPIV-3 fusion glycoprotein (see GenBank:ARV77784.1,539 aa, https: / / www.ncbi.nlm.nih.gov / protein / ARV77784.1). In some embodiments, the HPIV-3-targeting fusion peptide inhibitor is an HRC peptide derived from the HPIV-3 fusion glycoprotein. In preferred embodiments, the fusion peptide inhibitor contains the same amino acid sequence as the amino acid sequence of residues 449V to 484I of the HPIV-3 fusion glycoprotein in the standard N-to-C direction. 【0045】 In a preferred embodiment, the fusion peptide inhibitor targeting HPIV-3 has the amino acid sequence shown in SEQ ID NO: 1: Ac q -VALDPIDISIELNKAKSDLEESKEWIRRSNQKLDSI (sequence number 1) [where q is 0 or 1] contains. 【0046】 In other preferred embodiments, the fusion peptide inhibitor comprises an amino acid sequence that is a native or non-native mutant sequence of the amino acid sequence extending from residues 449V to 484I of the HPIV-3 fusion glycoprotein in the standard N-to-C direction. As used herein, the term “mutant sequence” is defined as a peptide having at least one amino acid deletion, addition, or substitution compared to the wild-type sequence, e.g., SEQ ID NO: 1 or other native sequences described herein. The mutant sequence preferably binds to a cognitive ligand of the wild-type sequence. For example, a peptide in which amino acids 1, 2, 3, 4, or 5 of SEQ ID NO: 1 are substituted can be used. Such substituted amino acids can preferably be selected from one or more corresponding amino acids in different paramyxovirus strains, as identified by sequence alignment. 【0047】 In a further preferred embodiment, the fusion peptide inhibitor targeting HPIV-3 is: SEQ ID NOs: 2-6 Acq -VALDPIDISIVLNKIKSDLEESKEWIRRSNKILDSI(Sequence ID 2)[where q is 0 or 1], Ac q -VALDPIDISIVLNKIKSQLEESKWEIRRSNKILDSI(sequence number 3)[where q is 0 or 1], Ac q -VALDPIDFSIVLNKIKSQLEESKWEIRRSNKILDSI(sequence number 4)[where q is 0 or 1], Ac q -VALDPIDISIVLNKIKSQLEESKEWIRRSNKILDSI(sequence number 5)[where q is 0 or 1], Ac q -VALDPIDFSIVLNKIKSQLEESKEWIRRSNKILDSI(sequence number 6)[where q is 0 or 1] It contains an amino acid sequence selected from the following. 【0048】 In some embodiments, the fusion peptide inhibitor targeting HPIV-3 is one of the amino acid sequences from SEQ ID NOs: 1-6. 【0049】 In some embodiments, the compound of the present invention has the structure shown in Figure 1A (also known as "DCOY3001"). 【0050】 In some embodiments, the compounds of the present invention have the structure shown in Figure 1B (also known as "DCOY3002"). 【0051】 In some embodiments, the compound of the present invention has the structure shown in Figure 1C (also known as "DCOY3003"). 【0052】 In some embodiments, the compound of the present invention has the structure shown in Figure 1D (also known as "DCOY3004"). 【0053】 In some embodiments, in addition to the HRC peptide for paramyxoviruses, the compound may include additional peptides that target other domains of the F protein, such as fusion peptides (FPs) or receptor-binding domains (RBDs). For example, one class is fusion inhibitory peptides (FIPs) that specifically target fusion peptides (FPs) of the F protein. Another class is synthetic peptides that mimic the RBDs of the HN protein. For example, a compound targeting RSV may include the HRC peptide for RSV and an RBD-targeting peptide such as 101F, which has been shown to inhibit RSV infection by interfering with the HN-F interaction. 【0054】 In some embodiments, the compounds of the present invention have anticoronavirus activity. Coronaviruses target human cells via the binding domain of spike proteins that bind to the human angiotensin-converting enzyme 2 (hACE2) receptor on host cells. The coronavirus spike (S) glycoprotein is a class I virus fusion protein on the outer membrane of the virion and plays a crucial role in viral infection by recognizing host cell receptors and mediating the fusion of the viral membrane with the cell membrane. Coronavirus entry into host cells is mediated by the transmembrane spike (S) glycoprotein, which forms a homotrimer protruding from the viral surface. S contains two functional subunits: one responsible for binding to the host cell receptor (S1 subunit) and the other responsible for the fusion of the viral membrane with the cell membrane (S2 subunit). 【0055】 S1 performs the function of receptor binding and contains an N-terminal signal peptide (SP), an N-terminal domain (NTD), and a receptor-binding domain (RBD). S2 functions in membrane fusion to facilitate entry into cells and contains a fusion peptide (FP) domain, an internal fusion peptide (IFP), two heptad repeat domains (HR1 and HR2), a transmembrane domain, and a C-terminal domain. 【0056】 After binding, the spike protein is activated by the host cell's transmembrane protease / serine subfamily member 2 (TMPRSS2), resulting in the virus fusing with the endosomal membrane for entry into the cell. Because the membrane fusion domain of the spike protein in coronaviruses is highly conserved, targeting membrane fusion could lead to durable and sustained therapeutic agents. 【0057】 The compound comprises a fusion peptide inhibitor that targets coronavirus, preferably HRC peptide. The compound may further comprise a spike-binding peptide. The compound may further comprise a target peptide, such as an ACE2-targeting peptide or a receptor-binding domain-targeting peptide. 【0058】 Each peptide is independently an HRC peptide and / or targeted peptide derived from the coronavirus spike protein, provided that at least one HRC peptide derived from the coronavirus spike protein is present. Preferred embodiments of the present invention utilize either native or wild-type peptides, but non-native peptides can also be used. For example, amino acids found in one or more variants (e.g., omicron variants) can be combined with native sequences from other viruses (e.g., deltaviruses). HRC peptides inhibit viral fusion, a crucial early step in the infection process. The wild-type HRC peptide is a conserved region of the spike protein, or S protein, throughout the coronavirus. The fusion region (HRC / HRN) and fusion mechanism of class I enveloped viruses, due to their conserved nature, make them ideal targets for developing pan-coronavirus inhibitors. 【0059】 The preferred wild-type HRC peptide is, Ac n -DISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL(Sequence ID 7) [In the formula, n is either 0 or 1] This includes the following sequence and its associated fragment. 【0060】 Regarding the HRC peptide of Sequence ID No. 1, the conventional amino acid numbering begins with D as 1150. 1151I, 1154I, and 1158V are at the hydrophobic interface before fusion, while 1159V is exposed. These amino acids stabilize the helix. When a conformational change occurs (e.g., release of FP by protease clipping), 1158V becomes exposed, and 1159V is located on the hydrophobic surface interacting with the HRN trimer. 1155N and 1176N are associated with N-linked glycosylation, which is conserved in coronaviruses. The first seven amino acids are associated with HRN binding. The "N-cap" region spans 1159V and 1171V. 1171V is a conserved hydrophobic substance in coronaviruses, stabilizing the HRC hydrocore and participating in HRN interactions. 【0061】 The hydrophobic core spans 1161I and 1175L and is helix both pre- and post-fusion. These isoleucine, leucine, and alanine are important for the folding and stability of the coiled coil. The C-cap region spans 1176N and 1185L. 1179L, 1182L, and 1185L are at the hydrophobic interface pre-fusion, while 1180I is exposed. These amino acids stabilize the helix. 1185Y may be involved in the hydrophobic packing between the three polypeptide chains in the trimer coiled coil. Upon conformational change (release of FPs by protease clipping), 1179L becomes exposed, and 1180I is located on the hydrophobic surface interacting with the HRN trimer. 1164E and 1184E form a salt bridge between HRC and HRN. Furthermore, 1159V, 1160N, 1171V, and 1180I have been shown to interact with HRN in the crystal structure. 【0062】 In certain embodiments, the peptide is Ac q -DISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL(Sequence ID 7) [In the formula, q is either 0 or 1] It is selected from mutant sequences of the wild-type HRC peptide that include the sequence and its binding fragment. 【0063】 For example, as shown in the alignment above, one or both of the underlined isoleucines can be substituted with leucine and / or methionine. The underlined alanine can be substituted with valine, leucine, or isoleucine. One or both of the underlined leucines can be independently substituted with isoleucine, tyrosine, alanine, or valine. Other conserved or non-conserved substitutions (lysine and glutamine or aspartic acid and glutamic acid) can be similarly selected, as shown, for example, in the sequence alignment above. In embodiments, amino acids conserved among 2, 3, 4, 5 or more coronaviruses (e.g., coronaviruses isolated from bats, or variants or mutants of SARS-CoV-2) remain conserved in non-natural HRC peptides as well. 【0064】 For example, the wild-type HRC sequence can contain one, two, three, four, five, six, seven, eight, nine, ten, or more additional amino acids naturally present in the S protein at its N-terminus and / or C-terminus. For example, glycine can be added to the N-terminus. Furthermore, the wild-type HRC peptide fragment can inhibit infection even if one, two, three, four, five, six, seven, eight, nine, or ten amino acids are deleted from the N-terminus and / or C-terminus. Typically, a total of ten or fewer amino acids are deleted overall. For example, it can be expected that inhibitory activity will be retained even if those ten amino acids at the C-terminus are deleted. 【0065】 In certain embodiments, modification to the wild-type sequence is desirable. For example, the use of one or more D-amino acids may improve the pharmacokinetics and half-life of the peptide. Therefore, in certain embodiments, the present invention includes peptides characterized by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more D-amino acids. Preferably, the corresponding L-amino acids in the wild-type sequence may become the D-amino acids. In certain embodiments, the D-amino acids are amino acids located at or near the protease degradation site (e.g., within 1, 2, or 3 amino acids). In certain embodiments, the D-amino acids are hydrophobic amino acids that participate in binding to the HRN peptide, preferably with higher affinity than the corresponding wild-type sequence. Alternatively, or in addition to the above, the D-amino acids are hydrophilic amino acids, such as lysine, aspartic acid, glutamic acid, or arginine. Alternatively, or in addition to the above, the D-amino acids can be selected from the seven amino acids at the N-terminus of SEQ ID NO: 1. The peptide improved by incorporating a D-amino acid is described in U.S. Patent Application No. 63 / 140,387, filed on 22 January 2021, which is incorporated herein by reference in its entirety. 【0066】 However, replacing one or more D-amino acids with corresponding L-amino acids can alter the peptide topology and affect its function. Therefore, preferred non-natural HRC peptides are retroinverted HRC peptides, or "RI HRC peptides." Retroinverted HRC peptides are preferably characterized by a binding affinity to their cognitive ligand at least about 50% of that of wild-type HRC peptides in standard binding assays, and reduced sensitivity to mammalian protease degradation. Retroinversion is defined as reversing the D-peptide sequence of a helical peptide, i.e., "flipping" both ends, thereby restoring the presentation of the side chains to the binding ligand or target. See Kim et al, Method to generate highly stable D-amino acid analogs of bioactive helical peptides using a mirror image of the entire PBS, PNAS, February 13, 2018, 115(7)1505-1510. This document is incorporated herein by reference in its entirety. Therefore, the non-natural peptides of the present invention may include peptides having the sequence of SEQ ID NO: 1, in which the amino acids are D-amino acids, and for example, the amino acids within a certain region are inverted from the N-terminus to the C-terminus. For example, the N-terminus can be subjected to retroinversion as shown in SEQ ID NOs: 8-14, where each D-amino acid is preceded by "d". Ac q -dVdAdLdDdP IDISIELNKAKSDLEESKEWIRRSNQKLDSI (Sequence ID 8), Ac q -dVdAdLdDdP IDISIVLNKIKSDLEESKEWIRRSNKILDSI (Sequence ID 9), Ac q -dVdAdLdDdP IDISIVLNKIKSQLEESKWEIRRSNKILDSI(sequence number 10), Ac q-dVdAdLdDdP IDFSIVLNKIKSQLEESKWEIRRSNKILDSI(Sequence ID 11), Ac q -dVdAdLdDdP IDISIVLNKIKSQLEESKEWIRRSNKILDSI(sequence number 12), Ac q -dVdAdLdDdP IDFSIVLNKIKSQLEESKEWIRRSNKILDSI(sequence number 13), and Ac q -dIdGdSdIdD NASVVN I QKE I DRLNEV A KN L NES L IDLQEL(sequence number 14)[where q is 0 or 1]. 【0067】 These examples give a single RI region of 5 amino acids. However, it is also possible to select only two amino acids (e.g., two N-terminal amino acids). For example, in the case of the HRC peptide targeting coronavirus, the RI region may span the hydrophobic core from 1160N to 1176N, or the C-cap region or a portion thereof. Alternatively, the entire peptide can be the RI peptide. Furthermore, it is possible to include two, three, or more RI regions. For example, both the N-terminal region and the C-cap region can be the RI region while retaining the hydrophobic core provided by the L-amino acids. 【0068】 For example, when screening HRC variant sequences using enantiomer phage display, the first D-peptide can be synthesized from an HRN coronavirus peptide or a first L-peptide. The first L-peptide may be a naturally occurring L-peptide or a peptide chimera. The method may further include screening for an HRC peptide or a second L-peptide that specifically binds to the first D-peptide, and then synthesizing a second D-peptide that is a mirror image of the second L-peptide. In one embodiment of the D-peptide screening method described herein, an N-trimer target can be first synthesized with D-amino acids to create a mirror image of a naturally occurring LN-trimer target. The DN-trimer target can be used in standard peptide-based screenings for identifying L-peptides that bind to the DN-trimer, such as phage display, ribosome display, and / or CIS display. The identified L-peptide can then be synthesized with D-amino acids. Due to the law of symmetry, the resulting D-peptide binds to the natural LN-trimer and therefore targets the N-trimer region of the coronavirus HRN intermediate, thereby inhibiting infection. This screening method is also described in Schumacher, et al., Identification of D-peptide ligands through mirror-image phage display, Science, 1996 Mar 29;271(5257):1854-7, which is incorporated herein by reference in its entirety. 【0069】 The hotspot residues of an HRC peptide can be identified by the crystal structure or NMR solution structure of the HRC peptide. For example, one, two, three, four, five, six, seven, eight, nine, ten or more amino acids selected from 1150D, 1151I, 1154I, 1155N, 1158V, 1159V, 1161I, 1164E, 1171V, 1175L, 1176L, 1179L, 1180I, 1182L, 1184E and / or 1185Y, such as one or more amino acids selected from 1159V, 1160N, 1163E, 1171V, 1180I, 1184E and / or 1185L of SEQ ID NO: 7, can be designated as hotspot residues. 【0070】 In some embodiments, the peptide includes a cell-targeting peptide. In several embodiments, the targeting peptide is selected from, but is not limited to, an ACE2-targeting peptide or a receptor-binding domain-targeting peptide. In several embodiments, the targeting peptide is an ACE2-targeting peptide. In several embodiments, the targeting peptide is a receptor-binding domain-targeting peptide. 【0071】 In some embodiments, the compound contains only one peptide, the peptide is an HRC peptide derived from the coronavirus spike protein. In embodiments, the compound contains two or more peptides, each peptide may be an HRC peptide derived from the coronavirus spike protein. In embodiments, the peptides may be selected from HRC peptides derived from the coronavirus spike protein and / or targeted peptides, provided that at least one HRC peptide derived from the coronavirus spike protein is present. Preferably, only one of the peptides is a targeted peptide, and the other peptides are HRC peptides derived from the coronavirus spike protein. 【0072】 In some embodiments, the compounds of the present invention have the structure shown in Figure 1E (also known as "DCOY104"). 【0073】 Automated Flow Chemistry There is a need for the ability to synthesize peptide conjugates using finely tuned, simple chemical reactions that are non-toxic to cells, can be performed rapidly, and can keep pace with changing disease or research needs. This invention addresses this need. In a broad embodiment, this disclosure provides functional modules of peptide conjugates and their subunits that are suitable for rapid automated flow chemistry. Automated flow peptide synthesis (AFPS), such as disclosed in U.S. Patent No. 10,683,325 (B2), is a solid-phase peptide synthesis system with feedback control that can give a high degree of control over individual coupling reactions to produce peptides and / or minimize side reactions. The method of the present invention provides an improvement over AFPS, enabling the accurate and rapid synthesis of compounds, and is therefore also called "one-shot synthesis" of multi-part compounds. The method of the present invention provides an opportunity to rapidly and accurately synthesize a wide variety of peptide conjugates that can be used for applications in prevention, diagnosis, and therapy. 【0074】 In some embodiments, the synthesis of compounds can be carried out in an AFPS system including a reactor comprising peptides immobilized on a solid support. For example, as shown in Figure 2, reagent reservoirs (1-6) can be located upstream of reactor (8) and fluidly connected to reactor (8). For example, the first reagent reservoir (1), the second reagent reservoir (2), and the third reagent reservoir (3) can be fluidly connected to the reactor via a first reagent channel, a second reagent channel, and a third reagent channel, respectively, which are connected to a delivery channel connected to the reactor. The reagent reservoirs contain at least some of the reagents necessary for compound synthesis. For example, the first reagent reservoir contains amino acids, the second reagent reservoir contains an activator (e.g., a uronium activator), and the third reagent reservoir contains a reagent including a membrane-immobilized portion such as cholesterol or a derivative thereof. The system may further include optional reagent reservoirs (e.g., 4, 5, or 6) which may contain spacer reagents such as PEG molecules. The system may further include an optional reagent reservoir (e.g., 4, 5, or 6) which may contain a deprotection reagent such as piperidine or trifluoroacetic acid. The system may further include an optional reagent reservoir (e.g., 4, 5, or 6) which may contain a base and / or an optional reagent. The system may also include an optional reagent reservoir (e.g., 4, 5, or 6) fluidly connected to the reactor, which may contain a solvent such as dimethylformamide (DMF) which can be used, for example, in a reagent removal step. The system may further include a reactor (7) which may optionally be configured to accelerate and / or facilitate one or more chemical reactions between a particular reagent and / or its reaction products, for example, by adjusting the reaction rate and / or reaction time. 【0075】 For simplicity, Figure 2 shows a single reservoir, but please understand that instead of a single reservoir, multiple reservoirs (for example, each containing different types of amino acids, different types of activators, different types of deprotective agents, different types of bases, etc.) can be used. 【0076】 During the amino acid addition step, a single additional amino acid residue can be incorporated into the peptide by adding amino acid (9) (i.e., a single amino acid residue can be added to the immobilized peptide so that a given peptide contains a single additional amino acid residue after the addition step). In some such embodiments, the peptide can be constructed by using a subsequent amino acid addition step to add amino acid residues individually until the desired peptide is synthesized. In some embodiments, two or more amino acid residues (e.g., in the form of peptides) can be added to a peptide immobilized on a solid support 10 (i.e., a peptide containing multiple amino acid residues can be added to a given immobilized peptide). The addition of peptides to immobilized peptides can be achieved by processes known to those skilled in the art (e.g., fragment condensation, chemical ligation). That is, adding amino acids to an immobilized peptide during the amino acid addition step can include adding a single amino acid residue to the immobilized peptide or adding multiple amino acid residues (e.g., as peptides) to the immobilized peptide. 【0077】 In some embodiments, the synthesis system includes flowing a fluid stream, for example, from a reagent reservoir to a reactor (e.g., 7, 8). For example, after a coupling reaction, a fluid stream containing a deprotection reagent may be flowed into the reactor. The fluid can exit the reactor through an efflux channel connected to the reactor. The efflux channel may be fluid-connected to a detection zone (e.g., 11). In certain embodiments, the efflux channel (e.g., 12) may not include a separation element (e.g., a size exclusion column, an affinity column) and / or may be connected to a detection zone. 【0078】 In some embodiments, a detection zone (e.g., 11) may include one or more electromagnetic radiation detectors. The detectors may measure the electromagnetic absorption and / or emission of one or more fluids leaving the reactor and generate one or more signals corresponding to the fluids. The signals may be transmitted to a unit (e.g., 13) that electrically communicates with the detectors. In some embodiments, the unit (e.g., 13) may be a controller configured to control one or more parameters of the system. In such embodiments, the controller may be operably associated with one or more components of the system (e.g., a temperature controller, a fluid flow source), and / or one or more processors for controlling the components of the system. For example, the controller may be operably associated with one or more processors to control flow rate, temperature, selection of reagent type, selection of reagent concentration, reaction time, selection of reagent ratio, addition of additives, or a combination thereof. Optionally, the controller may also be operably associated with other components, such as a user interface and an external communication unit (e.g., USB), and / or other components, as described in more detail below. The user interface may be used to display signals, to alert the user to problems related to specific responses, and / or to receive operational instructions from the user. 【0079】 System feedback control enables rapid and accurate synthesis of compounds. In some embodiments, feedback control is performed by detecting electromagnetic absorption and / or emission of the fluid flow in a detection zone located downstream of the reactor (e.g., 11 in Figure 2) during and / or immediately after the reaction step in the amino acid addition cycle (e.g., after the flow has left the reactor but before the next amino acid addition step) to generate a signal. At least in part, based on information derived from the signal (e.g., intensity and time components derived from the signal), one or more parameters of the system can be adjusted before and / or during the subsequent reaction in the reactor (e.g., a coupling reaction), and / or before the formation of the peptide in the reactor. Feedback can be used to adjust system parameters by controlling, for example, one or more pumps, vacuum devices, valves, temperature controllers, and / or other components. In some cases, feedback can determine a problem that has occurred or is occurring in the solid-phase peptide synthesis system, and a controller can send one or more signals to one or more components to adjust parameters in the whole or in part of the system. Alternatively, if corrective action cannot be taken, the controller may send one or more signals to one or more components to shut down the system. 【0080】 In some embodiments, detection during the reaction steps in the amino acid addition cycle may include detection from the start to the end of the reaction step, detection during at least a portion of the reaction step (e.g., at least about 20% of the total reaction time, at least about 30% of the total reaction time, at least about 40% of the total reaction time, at least about 50% of the total reaction time, at least about 60% of the total reaction time, or at least about 75% of the total reaction time, and less than about 100%), and / or detection during at least a portion of two or more reaction steps (e.g., a deprotection step), and / or continuous detection during the entire peptide synthesis. 【0081】 In some embodiments, both heating the amino acids and combining them with a base and / or activator can be performed before the amino acids come into contact with the immobilized peptide and for a relatively short period of time. Heating the amino acids may be performed before, during, and / or after the combination of flows. 【0082】 Reagents used in compound synthesis include protecting groups on, for example, primary amines of amino acids, the N-terminus and / or side chains of peptides, functional groups of linkers, subunits of linkers, or hydrophilic spacers. As used herein, the term “protecting group” is given in the ordinary sense of the art. A protecting group comprises a chemical moiety that binds to, or is configured to bind to, a reactive group (i.e., the group to be protected) within a molecule (e.g., a peptide), thereby preventing or otherwise inhibiting the reaction of the group to be protected. Protection can be achieved by attaching the protecting group to the molecule. In some embodiments, the side chains of amino acid residues in a peptide may contain protecting groups. Deprotection can occur when a protecting group is removed from the molecule, for example, by a chemical transformation that removes the protecting group. Generally, any protecting group known to those skilled in the art can be used. Non-limiting examples of protecting groups (e.g., n-terminal protecting groups) include fluorenylmethyloxycarbonyl (Fmoc), 4-methyltrityl (Mtt), tert-butyloxycarbonyl, allyloxycarbonyl (alloc), carboxybenzyl, and photosensitive protecting groups. In certain embodiments, the fixed peptide contains a fluorenylmethyloxycarbonyl protecting group. In some embodiments, the fixed peptide, B moiety, linker, or spacer may contain an Fmoc protecting group and / or an Mtt protecting group. 【0083】 As described elsewhere, activators may be used to activate or complete the activation of amino acids before exposing them to a fixed peptide. Any suitable activator may be used. In some embodiments, the activator includes carbodiimides, e.g., N,N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), etc. In certain embodiments, the activator includes uronium activators, e.g., O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU), 1-[(1-(cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylaminomorpholino)]uronium hexafluorophosphate (COMU), etc. 【0084】 As described elsewhere, peptides can be immobilized on a solid support. In general, any solid support can be used in any of the addition cycles described herein. Non-limiting examples of solid support materials include polystyrene (e.g., in resin forms such as microporous polystyrene resin, mesoporous polystyrene resin, macroporous polystyrene resin), glass, polysaccharides (e.g., cellulose, agarose), polyacrylamide resin, polyethylene glycol, or copolymer resins (e.g., polyethylene glycol, polystyrene, etc.). 【0085】 The polymer core B is designed to conform to the above method through automated flow chemistry so that the amino acids of the peptide, an optional linker, and a membrane-fixed portion are bonded to the polymer core B. In some embodiments, the solid support is preferably treated to present an amine group on its surface, so that the carboxylate group of lysine reacts with the amine group and is thereby fixed to the solid support, after which a coupling reaction takes place with the other subunits of the B portion, e.g., a second lysine or hydrophilic spacer, the amino acids of the peptide, the linker, and the membrane-fixed portion, and selective or total deprotection of the functional group can be performed before the coupling reaction or between the two coupling reactions. 【0086】 This invention provides a method for rapidly synthesizing peptide conjugates. 【0087】 In some embodiments, the method includes: forming a peptide fragment by flowing a fluid stream containing a deprotection reagent into a reactor after a coupling reaction between an amino acid and an amino acid residue immobilized on a solid support; generating a signal by detecting electromagnetic absorption and / or electromagnetic emission of the fluid stream in a detection zone located downstream of the reactor; and adjusting the parameters of the system, at least in part, based on intensity and time components derived from the signal, before peptide formation in the reactor, wherein the peptide contains a peptide fragment; optionally, flowing a fluid stream containing a hydrophilic polymer molecule into the reactor; optionally, flowing a fluid stream containing a deprotection reagent into the reactor after a coupling reaction between the peptide fragment and the hydrophilic polymer molecule; and flowing a fluid stream containing a reagent containing a membrane-immobilized portion into the reactor. Preferably, the peptide fragment contains a lysine residue at its C-terminus. Preferably, the hydrophilic polymer molecule is PEG or a derivative thereof. Preferably, the membrane-immobilized portion is cholesterol. 【0088】 In some embodiments, the method comprises: a) a coupling reaction to prepare a fixed N-protected diamino acid, wherein at least one N-protected diamino acid is bonded to a solid support having a surface amino group, the fixed N-protected diamino acid being bonded to the solid support via an amide bond and having two unbonded amine groups, with at least one amine group being protected; b) optionally a coupling reaction to prepare a fixed N-protected peptide, wherein a fixed N-protected diamino acid is reacted with at least one additional N-protected diamino acid in a C-to-N direction, the fixed N-protected peptide having 2 to 6 diamino acid residues in length and its C-terminus being bonded to a solid support; and c) optionally a coupling reaction to prepare a fixed conjugate framework, wherein the fixed N-protected diamino acid of step (a) or the fixed N-protected peptide of step (b) is reacted with a linker, the fixed conjugate framework comprising at least one N-protected diamino acid and at least one linker covalently bonded to each other at the amino group of the N-protected diamino acid, each linker independently PEG 2~40d) Reacting a PEG reagent comprising a chain and two reactive end groups, d) synthesizing an amino acid sequence, preferably in an automated flow peptide synthesis system (AFPS), wherein the C-terminus of the amino acid sequence is coupled to a primary amine of the immobilized portion by a coupling reaction, and the immobilized portion is selected from the immobilized N-protected diamino acid of step (a), the immobilized N-protected peptide of step (b), and the immobilized conjugate framework of step (c), e) optionally protecting the N-terminus of the amino acid sequence of step (d), wherein the protecting group is acyl or acetyl, f) protecting the primary amine The process includes: (a) deprotecting the protecting amine group in step (a) to form a product; (g) optionally reacting the primary amine in step (f) with a linker via a coupling reaction to form a product, wherein the linker is as defined in step (c); (h) reacting either the primary amine in step (f) or the reactive chain end of the linker in step (g) with a functional group via a coupling reaction to form a product, wherein the functional group is a target portion, a binding portion, or a film-fixing portion; and (i) cleaving the product of step (h) from a solid support. 【0089】 In some embodiments, the method involves: a) attaching at least one N-protected diamino acid, e.g., N-protected lysine, which is a diamino acid substituted with a protecting group on the amine, to a solid support having a surface amino group in order to prepare a fixed N-protected diamine containing a primary amine; b) reacting the fixed N-protected diamine with a diamino acid, e.g., lysine, or with an N-protected diamino acid, e.g., N-protected lysine, in order to prepare a fixed N-protected polyamine having one or more primary amines; c) optionally repeating step (b); d) optionally reacting each primary amine in the fixed N-protected polyamine of step (b) or step c) with a linker moiety; e) preferably an automated flow peptide synthesis system. (AFPS) comprises synthesizing an amino acid sequence bonded (preferably via the C-terminus of the amino acid sequence) to the fixed N-protected polyamine of step (b) or step (c), more preferably to the linker moiety of step (d), (f) optionally protecting each N-terminus of each amino acid sequence, for example, by substituting each primary amine with an acyl or acetyl group, (g) removing each protecting group to form a product having at least one primary amine, (h) optionally reacting each primary amine with the linker moiety, (i) reacting either the primary amine of the product of step (g) or the linker moiety of the product of step (h) with a functional group, and (j) cleaving the product of step (i) from the solid support. Preferably, the product prepared by this method has the structure: [ka] [In the formula, n and m are independent integers greater than or equal to 1, where m+n is preferably greater than or equal to 3.] Each L1 and L2 is independently the same or different linker. Each peptide comprises an amino acid sequence, preferably a therapeutic peptide. Each R is a functional group, X is a peptide containing two or more diamino acids. It has. 【0090】 In some embodiments, this method a) Structure: [ka] [In the formula, SS is a solid support, L is a linker, and PG is a protecting group.] To prepare a fixed N-protected diamine having the following characteristics, at least one N-protected diamino acid is bonded to a solid support having a surface amino group. b) Structure: [ka] [In the formula, each L is independently a linker, preferably the same linker, for example, an aliphatic linker, preferably a C4 alkylene.] To prepare a fixed N-protected polyamine having the following characteristics, a fixed N-protected diamine of formula (I) is reacted with a diamino acid. c) Depending on the case, the structure: [ka] [In the formula, each linker is independently a linker such as polyethylene glycol, and X is the reactive part.] To prepare a product having the above, the fixed N-protected polyamine of step (b) is reacted with the linker. d) Preferably, in an automated flow peptide synthesis system (AFPS), an amino acid sequence is synthesized for each reactive moiety X. e) Depending on the case, amino acid sequence [ka] [In the formula, each peptide is independently an amino acid sequence, and R is independently H or a protecting group, such as acyl or acetyl.] Protecting the N-terminus, f) Structure: [ka] To form a product having a primary amine, the protecting group PG is removed. g) Depending on the case, the structure: [ka] [In the formula, linker-X is as defined in process (c)] To form a product having the following characteristics, a primary amine is reacted with a linker. h) Structure: [ka] [In the formula, FG is a functional group, such as a targeting moiety or lipid, and m is 0 or 1.] To form a product having the above, either the primary amine of formula (e) or X of formula (f) is reacted with the functional group. i) Cutting the product of equation (g) from the solid support. Includes. 【0091】 In some embodiments, this method a) (Peptide-linker) n The method involves synthesizing a fixed amino acid sequence containing lysine, wherein each (peptide-linker) is linked to lysine directly via an amide bond or indirectly via a polyethylene glycol (PEG) group, and the amino acid sequence is linked to a solid support via an amide bond between the C-terminal lysine of the amino acid sequence and the solid support, and one -NH2 group of the C-terminal lysine is protected, preferably the amino acid sequence is of the formula [ka] [In the formula, [ka] [where m is a covalent bond between the amino acid sequence and the solid support, m is 0, 1, 2, 3, 4, or 5, and each p is an integer independently selected from the range 0 to 40] The act of composing, as represented by b) In some cases, the N-terminus of the amino acid sequence may be protected (e.g., by acetylation), c) Deprotection of the NH2 group of the C-terminal lysine, d) In some cases, reacting an amino acid sequence with a protective PEG reagent, wherein the protective PEG reagent covalently bonds to the amino acid sequence via an amide bond at the C-terminus, for example, if the protective PEG reagent is Fmoc-NH-PEG o The reaction is -CH2COOH, where O is an integer selected from 2 to 20. e) In some cases, the protected PEG reagent may be deprotected. f) Reacting an amino acid sequence with a reagent containing a membrane-fixed portion, wherein the membrane-fixed portion is covalently bonded to the amino acid sequence via an amide bond at the C-terminal carbon, preferably the membrane-fixed portion is a lipid, and the reagent containing the membrane-fixed portion is, for example, cholesteryl hydrogen succinate. g) Cutting the product obtained from step F from the solid support. Includes. 【0092】 In some embodiments, lysine immobilized on a solid support of the system can be represented by the following formula: [ka] Subsequently, the protecting group can be removed, deprotecting the protected amine group and enabling the coupling reaction described above. Examples of protected lysine reagents include, but are not limited to, Fmoc-Lys(Mmt)-OH, Fmoc-Lys(Dde)-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys-OH, and Fmoc-Lys(Fmoc)-OH. For example, in the method described herein, the C-terminal lysine bound to the solid support may preferably be derived from Fmoc-Lys(Mtt)-OH. The non-C-terminal lysine bound to the solid support may preferably be derived from Fmoc-Lys(Fmoc)-OH. 【0093】 In some embodiments, the amino acids for peptide synthesis, an optional linker and its subunits, the hydrophilic spacer of the B portion, and the membrane-fixed portion are each protected and deprotected independently. For example, Fmoc-NH-PEG-carboxylate can be used as a precursor for the hydrophilic spacer. 【0094】 In some embodiments, the peptide conjugates prepared by this method are compounds described herein. In some embodiments, the peptide conjugates prepared by this method have a structure selected from the general formula, formula (I), formula (II), and formula (III), where all variables are as defined above, including in all embodiments and preferred embodiments. Non-limiting examples of peptide conjugates prepared by this method are shown in Figures 1A-1E. 【0095】 In some embodiments, the peptide conjugates prepared by this method exhibit antiviral activity against SAR-Cov-2 mutant strains. The peptide conjugate preferably comprises the amino acid sequence of SEQ ID NO: 7 or 14. 【0096】 In some embodiments, the peptide conjugates prepared by this method have antiviral effects against paramyxovirus mutants, such as HPIV3 or its mutants. The peptide conjugate preferably contains one of the amino acid sequences of SEQ ID NOs: 1-6 and 8-13. 【0097】 Pharmaceutical composition The compositions of the present invention comprise compounds described herein and pharmaceutically acceptable carriers. For example, the compositions can be administered systemically or topically. The compositions can be administered, for example, orally, intravenously, intramuscularly, rectally, cutaneously, subcutaneously, topically, transdermally, sublingually, nasally, by inhalation, or vaginally. Accordingly, the compositions can take the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, ointments, liquids, osmotic delivery devices, suppositories, enemas, injections, implants, sprays, or aerosols. The compositions can be formulated according to conventional pharmaceutical practices (e.g., Remington: The Science and Practice of Pharmacy, 22 nd edition,2013,ed.LVAllen,Pharmaceutical Press,Philadelphia and Encyclopedia of Pharmaceutical Technology,4 th (See Edition, ed. J. Swarbrick, 2013, CRC Press, New York.) 【0098】 The compounds can be formulated in various ways known in the art. For example, if one or more of the compounds of the present invention exist, along with any additional biologically active substance as defined herein, the additional substance may be formulated together or separately. 【0099】 Each compound of the present invention can be formulated for controlled-release (e.g., sustained or metered) administration, either alone or in combination with one or more active agents described herein, as described in U.S. Patent Application Publications 2003 / 0152637 and 2005 / 0025765, each of which is incorporated herein by reference. For example, the compounds of the present invention can be incorporated into capsules or tablets administered to a patient, either alone or in combination with one or more of the biologically active agents described herein. 【0100】 Known controlled-release formulations in the art include sustained-release microparticles or nanoparticles for surgical insertion or implantation, insertion, injection or injection, such as specially coated pellets, polymer formulations or polymer matrices, as microspheres or microcapsules, where the sustained release of the active drug is achieved by sustained or controlled diffusion from the matrix and / or selective degradation of the coating of the preparation or selective degradation of the polymer matrix. Other formulations or vehicles for controlled, sustained, or immediate delivery of the active ingredient to a preferred localized site in the patient include, for example, lipid nanoparticles (LNPs), suspensions, emulsions, gels, liposomes, and any other suitable delivery vehicles or formulations known in the art that are acceptable for subcutaneous or intramuscular administration. 【0101】 Suitable biocompatible polymers can be used as controlled-release materials. The polymer material may include biocompatible biodegradable polymers, and in certain preferred embodiments, it is preferably a copolymer of lactic acid and glycolic acid. Preferred controlled-release materials useful in the formulations of the present invention include polyanhydrides, polyesters, copolymers of lactic acid and glycolic acid (preferably with a weight ratio of lactic acid to glycolic acid of 4:1 or less, i.e., 80% by weight or less of lactic acid to 20% by weight or more of glycolic acid), and polyorthoesters containing a catalyst or degradation-promoting compound, for example, at least 1% by weight of an anhydride catalyst, such as maleic anhydride. Examples of polyesters include polylactic acid, polyglycolic acid, and polylactic acid-polyglycolic acid copolymers. Other useful polymers include protein polymers, such as collagen, gelatin, fibrin and fibrinogen, and polysaccharides, such as hyaluronic acid. 【0102】 In further embodiments, the controlled-release material acting as a carrier for the compounds of the present invention may further include bioadhesive polymers, such as pectin (polygalacturonic acid), mucopolysaccharides (hyaluronic acid, mucin), or non-toxic lectins, or the material itself may be a bioadhesive, such as polyanhydride or polysaccharide, such as chitosan. In embodiments in which the biodegradable polymer includes a gel, one such useful polymer is a thermogelling polymer, such as polyethylene oxide, polypropylene oxide (PEO-PPO) block copolymer, such as BASF Wyandotte's PLURONIC® F127. 【0103】 Formulations for oral use include tablets containing the active ingredient as a mixture with non-toxic, pharmaceutically acceptable excipients. These excipients may include, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch, e.g., potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate), granulators and disintegrants (e.g., cellulose derivatives, e.g., microcrystalline cellulose, starch, e.g., potato starch, croscarmellose sodium, arginate, or alginic acid), binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol), as well as lubricants, flow enhancers, and anti-adhesion agents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients may include colorants, flavorings, plasticizers, water-retaining agents, buffers, and flavoring agents (e.g., hydroxypropyl methylcellulose, hydroxypropylcellulose). 【0104】 One or more of the compounds of the present invention may be mixed together in a tablet, capsule or other vehicle, or they may be separated. In one example, the compounds of the present invention are contained inside the tablet, and the biologically active substance is on the outside of the tablet, such that a substantial portion of the biologically active substance is released prior to the release of the compounds of the present invention. 【0105】 Formulations for oral use may also be provided as chewable tablets, or as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin, or olive oil). Powders, granules, and pellets may be prepared using the ingredients mentioned above in the sections on tablets and capsules, by conventional methods, for example, using a mixer, fluidized bed apparatus, or spray drying equipment. Formulations for oral use may also be provided as mouthwashes, oral sprays, gargles, or oral ointments, or oral gels. 【0106】 Dissolution or diffusion-controlled release can be achieved by a suitable coating of the compound in tablet, capsule, pellet or granular formulation, or by incorporating the compound into a suitable matrix. Controlled release coatings may include the coating materials mentioned above, as well as / or one or more of the following, for example: shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitosterate, ethylcellulose, acrylic resins, dl-polylactic acid, cellulose acetate / butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-hydroxymethacrylate, methacrylate hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycol. In controlled-release matrix formulations, the matrix material may also include, for example, hydrated methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbons. 【0107】 Liquid forms in which the compounds and compositions of the present invention can be incorporated for oral administration include aqueous solutions, appropriately flavored syrups, aqueous or oily suspensions, and flavored emulsions using edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar medicinal vehicles. 【0108】 Suitable formulations for parenteral administration (e.g., by injection) include aqueous or non-aqueous isotonic pyrogen-free sterile solutions (e.g., solutions, suspensions) in which the compound is dissolved, suspended, or otherwise provided (e.g., in liposomes or other microparticles). Such liquids may further contain other pharmaceutically acceptable components, such as antioxidants, buffers, preservatives, stabilizers, bacteriostatic agents, suspending agents, thickeners, and solutes to make the formulation isotonic with the blood (or other relevant body fluids) of the intended recipient. Examples of excipients include, for example, water, alcohol, polyols, glycerol, and vegetable oils. Examples of suitable isotonic carriers for use in such formulations include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Typically, the concentration of the compound in the liquid is about 1 ng / ml to about 10 ug / ml, for example, about 10 ng / ml to about 1 ug / ml. The formulations may be provided in sealed containers of unit dose or multi-dose form, such as ampoules and vials, and may also be stored in a freeze-dried state, requiring only the addition of a sterile liquid carrier, such as sterile water for injection, immediately before use. Immediate injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. 【0109】 The composition of the present invention may include a liquid vehicle suitable for intranasal administration. The vehicle is preferably an aqueous solution. More preferably, the vehicle is an aqueous solution containing a thickening agent and optionally one or more additional excipients, such as those that improve the stability and / or comfort of the formulation during administration. 【0110】 Various thickeners are known in the art. Examples of thickeners include hydrophilic polymers, such as polysaccharides, polysaccharide derivatives, proteins, and synthetic polymers. Examples include, but are not limited to, gum arabic, tragacanth gum, alginic acid, carrageenan, locust bean gum, guar gum, gelatin, hyaluronic acid, polyacrylate, polyacrylate / alkyl acrylate copolymer, polyvinyl alcohol, polyvinylpyrrolidone, starch, polypropylene glycol alginate, maltodextrin, and cellulose ether derivatives, such as methylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, and carboxymethylcellulose. Where possible, salt forms of any of the above substances are preferred. Preferred thickeners include hyaluronic acid, such as sodium hyaluronate; carboxymethylcellulose, such as sodium carboxymethylcellulose and calcium carboxymethylcellulose; methylcellulose; hydroxyethylcellulose; hydroxypropylmethylcellulose; and hydroxypropylcellulose. 【0111】 This composition may optionally include one or more additional excipients, such as those that enhance ease of administration, comfort to the subject, or stability of the composition. Suitable additional excipients include, but are not limited to, tonicity modifiers such as sodium chloride and dextrose, antioxidants such as butylated hydroxyanisole, buffers such as sodium bicarbonate, sodium citrate and sodium phosphate, preservatives such as benzalkonium chloride, ethanol, propylene glycol, benzoyl alcohol, phenethyl alcohol, chlorobutanol or methylparaben, pH adjusters such as hydrochloric acid, sulfuric acid, sodium hydroxide, surfactants such as polysorbate 80, polysorbate 20 and polyoxyl 400 stearate, chelating agents such as disodium EDTA, antioxidants, cosolvents such as ethanol, PEG400 and propylene glycol, penetration enhancers such as oleic acid, and water-retaining agents such as glycerin (S. Thorat, Sch.J.App.Med.Sci.2016,4(8D):2976-2985, D. Marx et al.). See al., IntechOpen, DOI:10.5772 / 59468 (available from intechopen.com / books / drug-discovery-and-development-from-molecules-to-medicine / intranasal-drug-administration-an-attractive-delivery-route-for-some-drugs). 【0112】 In one embodiment, the vehicle consists of sodium hyaluronate, aloe vera, allantoin, sodium chloride, sodium bicarbonate, glycerin, propylene glycol, benzalkonium chloride, and USP-grade purified water. Suitable vehicles are marketed by NeilMed under the trademark NASOGEL. 【0113】 The amount of the active substance in the composition may vary, for example, from about 0.5% by weight to about 25% by weight. 【0114】 The pH of the formulation is acceptable in the nasal cavity, preferably at least about 8.0. Buffers that can be used in this formulation include, but are not limited to, phosphates, tris(hydroxymethyl)methylamino]propanesulfonic acid, 2-(bis(2-hydroxyethyl)amino)acetic acid, and N-[tris(hydroxymethyl)methyl]glycine, and Alkaline Buffer (Seachem). 【0115】 A pharmaceutical composition suitable for intranasal or pulmonary administration, comprising a water-soluble solvent selected from the group consisting of propylene glycol, glycerin, polyethylene glycol, and combinations thereof. The composition may further comprise one or more of a polysaccharide gum, a nonionic surfactant, and a preservative. An exemplary polysaccharide gum is sclerotium gum. An exemplary surfactant is a poloxamer, such as poloxamer 188, but is not limited thereto. The preservative may be, for example, benzalkonium chloride. 【0116】 The composition is a dry powder and can be delivered by a dry powder inhaler, suspended in a propellant, or delivered by a nebulizer in an aqueous suspension or aqueous solution. 【0117】 For example, a solution or suspension of an active substance and a pulmonary excipient, such as lactose, can be spray-dried to form particles having a sufficient particulate fraction for delivery to the lungs or upper respiratory system. Alternatively, an aqueous solution or suspension can be sonicated to aerosolize it into droplets small enough to be inhaled, for example, by a nebulizer. 【0118】 Excipients include carbohydrates, such as monosaccharides, disaccharides, and polysaccharides. For example, monosaccharides such as dextrose (anhydrous and monohydrate), galactose, mannitol, D-mannose, sorbitol, and sorbose; disaccharides such as lactose, maltose, sucrose, and trehalose; trisaccharides such as raffinose; and other carbohydrates such as starch (hydroxyethyl starch), cyclodextrin, and maltodextrin. Other excipients suitable for use in the present invention are known in the art, including amino acids, as disclosed in WO95 / 31479, WO96 / 32096, and WO96 / 32149. Furthermore, mixtures of carbohydrates and amino acids are also within the scope of the present invention. It is also conceivable to include both inorganic substances (e.g., sodium chloride), organic acids and their salts (e.g., carboxylic acids and their salts, such as sodium citrate, sodium ascorbate, magnesium gluconate, sodium gluconate, tromethamine hydrochloride, etc.) and buffers. 【0119】 The composition may be used in the form of a dry powder or in the form of a stabilized dispersion system containing a non-aqueous phase. Therefore, the dispersion system or powder of the present invention may be used in conjunction with metered-dose inhalers (MDIs), dry powder inhalers (DPIs), nebulizers, nebulizers, or liquid dose instillation (LDI) techniques to provide effective drug delivery. With regard to inhalation therapy, it will be understood by those skilled in the art that the hollow and porous microparticles of the present invention are particularly useful in DPIs. Conventional DPIs comprise a powder formulation and a device that delivers a predetermined dose of the drug, either alone or blended with lactose carrier particles, as a dry powder aerosol for inhalation. 【0120】 The pharmaceutical product is easily dispersed into loose particles and formulated in such a way that the aerodynamic median mass diameter of the powder is characteristically within the range of approximately 0.5 to 10, preferably approximately 0.5 to 5.0 microns (MMAD). 【0121】 As discussed above, the stabilized dispersion systems disclosed herein can also be administered by aerosolization, for example, using a metered-dose inhaler, into the nasal cavity or pulmonary airway of a patient. MDIs are well known in the art and can be readily used for administering the dispersion systems according to this application without excessive experimentation. Respiratory-actuated MDIs, along with other types of improvements already developed or to be developed in the future, are assumed to be within the scope as they are compatible with the stabilized dispersion systems and the present invention. However, it should be emphasized that in preferred embodiments, the stabilized dispersion systems can be administered by MDI using several different routes, including, but not limited to, topical, nasal, pulmonary, or oral administration. It will be understood by those skilled in the art that such routes are well known and that drug administration and administration procedures for the stabilized dispersion systems of the present invention can be readily found. 【0122】 Along with the embodiments described above, the stabilized dispersion system of the present invention may be used in conjunction with a nebulizer disclosed in PCT WO99 / 16420 to provide an aerosolized pharmaceutical product that can be administered to the pulmonary airways of patients requiring it, the disclosure of which is incorporated herein by reference in its entirety. Nebulizers are well known in the art and can be readily used for administering the dispersion system according to this application without excessive experimentation. Respiratory-operated nebulizers, including other types of improvements already developed or to be developed in the future, are also assumed to be compatible with and within the scope of the stabilized dispersion system and the present invention. 【0123】 It will be understood that, along with DPI, MDI, and nebulizers, the stabilized dispersion system of the present invention may be used in conjunction with liquid dose infusion techniques, i.e., LDI techniques, as disclosed, for example, in WO99 / 16421, which is incorporated herein in its entirety by reference. In liquid dose infusion, the stabilized dispersion system is administered directly to the lungs. In this respect, direct pulmonary administration of bioactive compounds is particularly effective in treating disorders in which the effectiveness of intravenous drug delivery is reduced, in particular, due to poor vascular circulation in the diseased portion of the lungs. With respect to LDI, the stabilized dispersion system is preferably used in conjunction with partial or total liquid ventilation. Furthermore, the present invention may further include introducing a therapeutically beneficial amount of a physiologically acceptable gas (such as nitric oxide or oxygen) into the pharmaceutical microdispersion system before, during, or after administration. 【0124】 How to use The present invention also includes methods of using the compositions of the present invention to treat or prevent an infection in subjects requiring treatment or prevention of the infection. The method comprises the step of administering an effective amount of the composition to the subject. The infection may be an infection of the gastrointestinal tract or the upper or lower respiratory tract, such as the common cold, influenza, respiratory syncytial virus infection, severe acute respiratory syndrome, Middle East respiratory syndrome, COVID-19, or a disease caused by another emerging zoonotic virus, such as zoonotic coronavirus. In certain embodiments, the methods of the present invention treat viral respiratory infections such as paramyxovirus (HPIV-3) or SARS-CoV-2 (COVID-19) respiratory infections. 【0125】 In some embodiments, the present invention provides a method for treating respiratory infections caused by paramyxoviruses or their variants in subjects requiring treatment. The method involves administering an effective amount of the formula: (peptide-linker) to the subject. nThe treatment involves administering a compound having a -B-membrane-fixed portion [wherein each peptide is independently an HRC peptide or a targeted peptide, provided that at least one peptide is an HRC peptide, each linker is independently a bivalent linking portion, B is a polyvalent portion comprising one or more lysines and optionally one or more spacers, the membrane-fixed portion is a lipid, and n is an integer selected from 1, 2, 3 or more]. Preferably, the paramyxovirus is HPIV-3. 【0126】 In some embodiments, the compound comprises a targeted peptide. The targeted peptide is an RSV fusion protein targeted peptide, an RSV-binding protein targeted peptide, an NiV / HeV-binding protein targeted peptide, or a receptor-binding peptide. Alternatively, the targeted peptide is a receptor-binding peptide selected from CD169-binding peptide, CD150-binding peptide, nectin-4-binding peptide, CX3CR1-binding peptide, heparin-binding peptide, chondroitin sulfate-binding peptide, dermatan sulfate-binding peptide, ephrin-B2-binding peptide, and ephrin-B3-binding peptide. 【0127】 In some embodiments, the present invention provides a method for treating respiratory infections caused by SAR-Cov-2 mutant strains in subjects requiring treatment of said infection. In some embodiments, the method involves administering an effective amount of the formula: (peptide-linker) to the subject. n The treatment involves administering a compound having a -B-membrane-fixed moiety [wherein each peptide is independently an HRC peptide or a targeted peptide, provided that at least one peptide is an HRC peptide; each linker is independently a bivalent linking moiety; B is a polyvalent moiety comprising one or more lysines and optionally one or more spacers; the membrane-fixed moiety is a lipid; and n is an integer selected from 1, 2, 3 or more]. Preferably, the SAR-Cov-2 mutant strain comprises at least five mutations, the at least five mutations independently located in the spike protein S1 subunit or S2 subunit or a combination thereof. 【0128】 In several embodiments, the compound comprises a targeted peptide and one or more HRC peptides. In several embodiments, the targeted peptide is an ACE2 targeted peptide or a receptor-binding domain peptide. In several embodiments, the hydrophobic moiety is cholesterol. 【0129】 In some embodiments, at least five mutations are independently located in the N-terminal domain (NTD), receptor-binding domain (RBD), fusion peptide (FP) domain, heptad repeat 1 (HR1) domain, or a combination thereof. In some embodiments, at least five mutations are independently selected from said mutations. 【0130】 In some embodiments, at least five mutations are independently selected from at least five mutations derived from the SAR-Cov-2 alpha mutant, at least five mutations derived from the SAR-Cov-2 beta mutant, at least five mutations derived from the SAR-Cov-2 delta mutant, or at least five mutations derived from the SAR-Cov-2 omicron mutant. In multiple embodiments, at least five mutations are independently selected from at least five mutations derived from the SAR-Cov-2 alpha mutant. In multiple embodiments, at least five mutations are independently selected from at least five mutations derived from the SAR-Cov-2 beta mutant. In multiple embodiments, at least five mutations are independently selected from at least five mutations derived from the SAR-Cov-2 delta mutant. In multiple embodiments, at least five mutations are independently selected from at least five mutations derived from the SAR-Cov-2 omicron mutant. 【0131】 In some embodiments, the mutant strain contains at least 10 mutations. In further embodiments, the mutant strain contains at least 15 mutations. In yet another embodiment, the mutant strain contains at least 20 mutations. 【0132】 In some embodiments, the SARS-CoV-2 mutant strain includes at least one mutant strain selected from B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma), B.1.617.2 (delta), B.1.429 / B.1.427 (epsilon), B.1.617.1 (kappa), B.1.525 (eta), B.1.526 (iota), P.3 (theta), P.2 (zeta), and B.1.1.529 (omicron). 【0133】 In some embodiments, the SARS-CoV-2 mutant strain includes at least one mutant strain selected from A.1-A.6, B.3-B.7, B.9, B.10, B.13-B.16, B.2, B.1 lineage, P.1, P.2, P.3, and R.1. 【0134】 In some embodiments, the B.1 system is, but is not limited to, B.1, B.1.1, B.1.1.7, B.1.2, B.1.5~B.1.72, B.1.9, B.1.13, B.1.22, B.1.26, B.1.37, B.1.3~B.1.66, B.1.177, B.1.24, which have E484K. 3, including at least one of B.1.313, B.1.351, B.1.427, B.1.429, B.1.525, B.1.526, B.1.526.1, B.1.526.2, B.1.617, B.1.617.1, B.1.617.2, B.1.617.3, B.1.619, B.1.620, and B.1.621. 【0135】 In some embodiments, administration is achieved using an intranasal spray, inhaler, or nebulizer. 【0136】 In some embodiments, the compound is administered in combination with at least one other antiviral agent or antiviral treatment. 【0137】 A subject, preferably a human, may be an individual diagnosed with an infectious disease and may be symptomatic, pre-symptomatic, asymptomatic, or asymptomatic, or at risk of developing an infectious disease. For example, a subject may be at risk of developing a viral respiratory infection due to direct or indirect exposure to or potential exposure to a virus (e.g., RSV, HPIV-3, SARS-CoV-2, or their variants), or, for example, exposure to an infected individual or a virus-contaminated vector. A subject may be a resident or visitor of an area where a viral respiratory infection has been identified, for example, a family member of an infected person, or a person working in a healthcare setting caring for an infected person. In certain embodiments, a subject at risk of infection may be asymptomatic and have tested negative for the presence of the virus before initiating treatment. In specific cases, a subject may be at risk of developing a condition resulting from exposure to the virus, for example, from respiratory droplets or aerosols from an infected person and / or contact with a contaminated vector, for example, at risk of developing COVID-19 due to exposure to the SARS-CoV-2 virus. In a further aspect, the subject is suffering from a condition related to a viral respiratory infection. 【0138】 In certain embodiments of the method of the present invention, the subject is suffering from another disease or condition, such as chronic obstructive pulmonary disease (COPD) or ulcerative colitis, which may be exacerbated by an infection. 【0139】 The composition is preferably administered to the subject before the subject becomes symptomatic (e.g., before the onset of symptoms) or at the time symptoms appear. The composition can be administered in various dosing schedules. For example, the composition can be administered once or multiple times over a day or several days. In certain embodiments, the composition is administered once or multiple times per day over a period of 1 to 10 days. In certain embodiments, the composition is administered once or multiple times per day until the subject becomes asymptomatic and / or tests negative for the virus. 【0140】 The composition can be administered intranasally using standard methods and devices (see D. Marx et al., IntechOpen, DOI:10.5772 / 59468, available from https: / / www.intechopen.com / books / drug-discovery-and-development-from-molecules-to-medicine / intranasal-drug-administration-an-attractive-delivery-route-for-some-drugs). For example, the composition can be administered intranasally as droplets or as an aerosol spray using an aerosol bottle or multi-dose spray pump capable of providing a uniformly measured dose. The volume per dose can vary, but is typically about 50 to about 150 μl. The desired volume depends on the desired dose of the active ingredient and the concentration of the active ingredient in the composition. 【0141】 If delivery to the pulmonary system or lungs is desired, it may be effective to aerosolize a low-concentration solution of the active ingredient over a long period of time, for example, overnight. 【0142】 Combination therapy The compounds or compositions described herein can be administered concurrently with other active substances and treatments. 【0143】 In some embodiments, other active ingredients include, but are not limited to, antibodies against paramyxoviruses, such as HPIV-3. For example, the protective antibody PI3-E12 against HPIV-3 is described in Boonyaratanakornkit et al., "Protective antibodies against human parainfluenza virus type 3 infection", mAbs, Volume 13, 2021, Issue 1, https: / / doi.org / 10.1080 / 19420862.2021.1912884. 【0144】 In several embodiments, other active ingredients include, but are not limited to, antibodies against SARS-CoV-2. Suitable antibodies are described, for example, in US2022 / 0017604, US2022 / 0017614, US2021 / 0403550, US2021 / 0395345, US2021 / 0403537, US2021 / 0388066, US2021 / 0388065, US2021 / 0347859, or US2021 / 0309733, which are incorporated herein by reference. In some embodiments, the antibody is a monoclonal antibody, such as casirivimab, imdevimab, bamranivimab, or etesevimab. In several embodiments, the antibody is a monoclonal antibody therapy, such as casirivimab + imdevimab, bamranivimab, or bamranivimab + etesevimab. 【0145】 The active substances and compositions of the present invention are also intended to be used in conjunction with general medical care provided to patients with viral infections, such as parenteral fluids (including dextrose saline and Ringer's lactate solution) and nutrition, antibiotics (including metronidazole and cephalosporin antibiotics, such as ceftriaxone and cefuroxime) and / or antiviral prophylaxis, antipyretics (such as acetaminophen) and analgesics, antiemetics (such as metoclopramide) and / or antidiarrheals, vitamin and mineral supplements (such as vitamin K and zinc sulfate), anti-inflammatory agents (such as ibuprofen), analgesics, and drug therapies for other common diseases in the patient population (such as artemether, artesunate-mefantrine combination therapy), quinolone antibiotics, such as ciprofloxacin, macrolide antibiotics, such as azithromycin, cephalosporin antibiotics, such as ceftriaxone, or aminopenicillins, such as ampicillin, or drug therapies for bacterial dysentery. 【0146】 Combination therapy may be administered as a concurrent regimen or a sequential regimen. When administered sequentially, the combination may be administered in two or more doses. 【0147】 Co-administration of the compound of the present invention with one or more other active therapeutic agents generally refers to simultaneous or sequential administration of the compound of the present invention and one or more other active therapeutic agents in such a manner that therapeutically effective amounts of both the compound of the present invention and one or more other active therapeutic agents are present in the patient's body. 【0148】 Co-administration includes the administration of a unit dose of the compound of the present invention before or after the administration of one or more other active therapeutic agents in a unit dose, for example, within seconds, minutes or hours after the administration of one or more other active therapeutic agents, and / or as part of the same treatment regimen. For example, a unit dose of the compound of the present invention may be administered first, followed by a unit dose of one or more other active therapeutic agents within seconds, minutes or days. Alternatively, a unit dose of one or more other therapeutic agents may be administered first, followed by a unit dose of the compound of the present invention within seconds, minutes or days. In some cases, it may be preferable to administer a unit dose of the compound of the present invention first, followed by a unit dose of one or more other active therapeutic agents several hours later (e.g., 1 to 12 hours). In other cases, it may be preferable to administer a unit dose of one or more other active therapeutic agents first, followed by a unit dose of the compound of the present invention several hours later (e.g., 1 to 12 hours). 【0149】 Combination therapy can produce a "synergistic effect," meaning that the combined effect achieved when active ingredients are used together is greater than the sum of the effects obtained when those compounds are used individually. 【0150】 As used herein, the words "a" and "an" shall encompass one or more unless otherwise specified. For example, the term "an agent" shall encompass both a single agent and a combination of two or more agents. 【0151】 As used herein, the terms “to treat” or “to treat” apply to the treatment of a disease or condition of interest in a mammal, preferably a human, having a disease or condition of interest (e.g., a respiratory infection), and include, for example, preventing or delaying the onset of the disease or condition in a mammal (especially if such a mammal is at risk of developing the disease but is not yet symptomatic and / or has not yet been diagnosed with the disease); inhibiting the disease or condition, i.e., stopping its onset; mitigating the disease or condition, i.e., causing a regression of the disease or condition; and / or stabilizing the disease or condition. Treatment includes alleviating or reducing the severity of the symptoms of the disease or condition and / or inhibiting further progression or worsening of those symptoms. Treatment also includes shortening the time course and / or severity of the disease or condition compared to the expected or past time course and / or severity of the disease. 【0152】 As used herein, the term “prevent” means preventing the development of the clinical symptoms of a disease or condition, and includes inhibiting the occurrence of a viral infection in a subject who is potentially exposed to or susceptible to a viral infection but has not yet experienced or presented any symptoms of such infection. 【0153】 The “effective amount” or “therapeutic effective amount” of a compound or composition described herein means an amount of the compound sufficient to achieve a particular effect or result, and / or to prevent or treat a disease or condition and / or its symptoms, thereby, for example, alleviating the symptoms associated with the disorder or condition in whole or in part, stopping or delaying the further progression or worsening of those symptoms, or preventing or providing prevention of the disorder or condition. Specifically, “effective amount” and “therapeutic effective amount” include the antiviral amount of the compound of the present invention or the composition described herein (either alone or in combination with another active agent). 【0154】 example Example 1. "All-on-Resin" One-Shot Synthesis The compounds described in this application were synthesized using the automated chemistry platform described in Hartrampf et al. "Synthesis of proteins by automated flow chemistry". Science 2020, Vol 368, Issue 6494, 980-987. DOI:10.1126 / science.abb2491. As an example, the synthesis of DCOY104 is shown below. [ka] 【0155】 compound 1 DISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGSGS 41 G- 42 Peg4)2- 43 K- 44 K-NH2 was synthesized using an automated flow peptide synthesizer (AFPS) with PyAOP as the activator and DIEA as the base, and Fmoc-based solid-phase peptide synthesis from the C-terminus to the N-terminus, in the formula, 44 K is FmocLys(Mtt)OH, 43 K is FmocLys(Fmoc)OH, 42 Peg4 was Fmoc-NH-PEG4-CH2COOH. After extraction from AFPS, the N-terminus was acetylated, and then... 44The Mtt side-chain protecting group on K was selectively deprotected and subsequently coupled with Fmoc-NH-PEG4-CH2COOH. The Fmoc group was deprotected, and the Peg4-amine was coupled with cholesteryl hydrogen succinate. Overall deprotection was performed by exposing the resin to TFA (94%), water (2.5%), EDT (2.5%), and TIPS (1%) for 1 hour. After 1 hour, the resin was filtered and then washed again with TFA. Cooled ether was added to the combined filtrate to precipitate the desired final product. This precipitate was purified by reverse-phase high-performance liquid chromatography to obtain the desired purity. The final product was confirmed by LC-MS. 【0156】 Example 2. In vitro assay Inhibition of virus-induced cytopathic effects (CPE) and cell viability after human respiratory syncytial virus replication in Hep2 cells were measured using XTT tetrazolium dye. 3 Cells (100 / well) were seeded into a 96-well flat-bottom tissue culture plate and allowed to adhere overnight. After overnight incubation, diluted test compounds and viruses, diluted to a predetermined titer to induce 85%–95% cell death on post-infection day 6, were added to the plate. After incubation at 37°C and 5% CO2 for 6 days, cell viability was measured by XTT staining. The optical density of the cell culture plate was determined spectrophotometrically at 540nm and 650nm using Softmax Pro 4.6 software. The CPE reduction rate (%) of the virus-infected wells and the cell viability (%) of the uninfected drug control wells were calculated and EC was analyzed using 4-parameter curve-fit analysis. 50 Value and TC 50 The values ​​were determined. The results are shown in Table 1. [Table 1] [Table 2] 【0157】 While the present invention has been specifically illustrated and described with reference to its preferred embodiments, it will be understood by those skilled in the art that various modifications in form and detail can be made without departing from the scope of the invention as encompassed in the appended claims. 【0158】 The patents and scientific literature referenced herein establish knowledge available to those skilled in the art. All U.S. patents and published or unpublished U.S. patent applications cited herein are incorporated herein by reference. All published foreign patents and foreign patent applications cited herein are incorporated herein by reference. All other published references, documents, manuscripts and scientific literature cited herein are incorporated herein by reference. All relevant teachings of all patents, published applications and references cited herein are incorporated herein by reference in their entirety.

Claims

[Claim 1] formula: (Peptide-linker) ｎ -B- Membrane fixing part [In the formula, each peptide is independently an HRC peptide or a targeted peptide, provided that at least one peptide is an HRC peptide; each linker is independently a divalent linking moiety; B is a polyvalent moiety comprising one or more diaminoliphatic acids, preferably lysine, and optionally one or more spacers; the membrane-fixed moiety is a lipid; and n is an integer selected from 1, 2, 3 or more.] A compound having the following properties. [Claim 2] The compound according to claim 1, wherein B contains 1, 2, 3, 4, 5 or 6 lysines. [Claim 3] The compound according to claim 1 or 2, wherein the spacer is independently selected from polyethylene glycol (PEG) derivatives. [Claim 4] A compound according to any one of claims 1 to 3, comprising a targeted peptide and one or more HRC peptides. [Claim 5] The compound according to any one of claims 1 to 4, wherein the HRC peptide is a paramyxovirus HRC peptide. [Claim 6] The compound according to any one of claims 1 to 5, wherein the HRC peptide is RSV HRC peptide, coronavirus HRC peptide, or HPIV HRC peptide. [Claim 7] The compound according to any one of claims 1 to 6, wherein the film-immobilized portion is selected from cholesterol, fatty acids, long-chain alkyl chains, 3β-cholesterylamine, 3β-cholesterylthiol, cholesteryl analogs and derivatives, 3β-cholesterylamine type molecules, and the above (preferably cholesterol) in the form of dimers, trimers or oligomers. [Claim 8] The aforementioned linker is AAY, EAAK, (GGGGS) ３ (GGGGS) ２ The compound according to any one of claims 1 to 7, wherein the compound is GGGGS or GSGSG. [Claim 9] The compound according to any one of claims 1 to 7, wherein the linker is a GS linker. [Claim 10] The compound according to any one of claims 1 to 9, wherein the linker is GSGSG. [Claim 11] Formula (III): 【Chemistry 1】 [In the formula, m is 0, 1, 2, 3, 4, or 5, and each p is an integer independently selected from the range 0 to 40.] The compound according to claim 1, represented by [the given expression]. [Claim 12] The compound according to claim 10, wherein each p is an integer independently selected from the range of 2 to 20, and the membrane-fixed portion contains cholesterol. [Claim 13] At least one peptide is a wild-type HRC peptide or a mutant thereof, and the wild-type HRC peptide is Ac ｑ -VALDPIDISIELNKAKSDLEESKEWIRRSNQKLDSI (Sequence ID 1) and Ac ｑ -DISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL (Sequence ID 7) [wherein q is 0 or 1] A compound according to any one of claims 1 to 12, comprising an amino acid sequence selected from. [Claim 14] At least one peptide, Ac ｑ -VALDPIDISIELNKAKSDLEESKEWIRRSNQKLDSI (Sequence ID 1), Ac ｑ -VALDPIDISIVLNKIKSDLEESKWIRRSNKILDSI (Sequence ID 2), Ac ｑ -VALDPIDISIVLNKIKSQLEESKWEIRRSNKILDSI (Sequence ID 3), Ac ｑ -VALDPIDFSIVLNKIKSQLLESKWEIRRSNKILD SI (SEQ ID NO: 4), Ac ｑ -VALDPIDISIVLNKIKSQLEESKEWIRRSNKILDSI (Sequence ID 5), Ac ｑ -VALDPIDFSIVLNKIKSQLEESKEWIRRSNKILDSI (Sequence ID 6), Ac ｑ -DISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL (Sequence ID 7), Ac ｑ -dVdAdLdDdP IDISIELNKAKSDLEESKEWIRRSNQKLDSI (Sequence ID 8), Ac ｑ -dVdAdLdDdP IDISIVLNKIKSDLEESKWIRRSNKILDSI (Sequence ID 9), Ac ｑ -dVdAdLdDdP IDISIVLNKIKSQLEESKWEIRRSNKILDSI (Sequence ID 10), Ac ｑ -dVdAdLdDdP IDFSIVLNKIKSQLEESKWEIRRSNKILDSI (Sequence ID 11), Ac ｑ -dVdAdLdDdP IDISIVLNKIKSQLEESKEWIRRSNKILDSI (Sequence ID 12), Ac ｑ -dVdAdLdDdP IDFSIVLNKIKSQLEESKEWIRRSNKILDSI (Sequence ID 13), and Ac ｑ -dIdGdSdIdD NASVVNIQKEIDRLNEVAKNLNESLIDLQEL (Sequence ID 14) A compound according to any one of claims 1 to 13, comprising an amino acid sequence selected from the above. [Claim 15] The compound according to any one of claims 11 to 14, wherein m is 0, 1, or 2. [Claim 16] The compound according to any one of claims 11 to 15, wherein all of the aforementioned peptides are the same and include an amino acid sequence selected from SEQ ID NOs. 1 to 12. [Claim 17] A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 and a pharmaceutically acceptable carrier. [Claim 18] A method for treating or preventing a respiratory infection associated with human parainfluenza virus (HPIV) in a subject requiring treatment or prevention, comprising: an effective amount of the formula: (Peptide-linker) ｎ -B- Membrane fixing part [In the formula, each peptide is independently an HPIV HRC peptide or a targeted peptide, wherein at least one peptide is an HPIV HRC peptide; each linker is independently a divalent linking portion; B is a polyvalent portion containing one or more lysines; the membrane-immobilized portion is a lipid; and n is an integer selected from 1, 2, 3 or more.] A method comprising administering a compound having [a certain characteristic]. [Claim 19] The method according to claim 18, wherein the HPIV is HPIV-1, HPIV-2, HPIV-3, or HPIV-4. [Claim 20] The method according to claim 18 or 19, wherein the HPIV is HPIV-3. [Claim 21] The method according to any one of claims 18 to 20, wherein the HPIV HRC peptide comprises an amino acid sequence selected from SEQ ID NOs: 1 to 6 and 8 to 13. [Claim 22] The method according to any one of claims 18 to 21, wherein the hydrophobic portion is cholesterol. [Claim 23] The aforementioned compound has the formula: 【Chemistry 2】 [In the formula, the linker is GSGSG, the peptide has an amino acid sequence selected from SEQ ID NOs: 1 to 14, and each p is an integer independently selected from 2 to 20.] The method according to any one of claims 18 to 22, as represented by [the specified method]. [Claim 24] The method according to any one of claims 18 to 23, wherein the administration is achieved using an intranasal spray, inhaler, or nebulizer. [Claim 25] A method for treating or preventing a respiratory infection associated with a SARS-CoV-2 variant in a subject requiring treatment or prevention, comprising: an effective amount of the formula: (Peptide-linker) ｎ -B- Membrane fixing part [In the formula, each peptide is independently an HRC peptide or a targeted peptide of the SARS-Cov-2 mutant strain, wherein at least one peptide is an HRC peptide of the SARS-Cov-2 mutant strain; each linker is independently a bivalent linking portion; B is a polyvalent portion containing one or more lysines; the membrane-fixed portion is a lipid; and n is an integer selected from 1, 2, 3 or more.] A method comprising administering a compound having [a certain characteristic]. [Claim 26] The method according to claim 25, wherein the targeted peptide is an ACE2-targeted peptide or a receptor-binding domain peptide. [Claim 27] The method according to claim 25 or 26, wherein the HRC peptide comprises the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO:

14. [Claim 28] The method according to any one of claims 25 to 27, wherein the hydrophobic portion is cholesterol. [Claim 29] The aforementioned compound has the formula: 【Transformation 3】 [In the formula, the linker is GSGSG, the peptide has the amino acid sequence of SEQ ID NO: 7 or 14, and each p is an integer independently selected from 2 to 20.] The method according to any one of claims 25 to 28, as represented by [the specified method]. [Claim 30] The method according to any one of claims 25 to 29, wherein the administration is achieved using an intranasal spray, inhaler, or nebulizer. [Claim 31] a) A step to prepare a fixed N-protected diamine containing a primary amine, comprising bonding at least one N-protected diamino acid, such as N-protected lysine, which is a diamino acid substituted with a protecting group on the amine, to a solid support having a surface amino group. b) A step of reacting the fixed N-protected diamine with a diamino acid, such as lysine, or with an N-protected diamino acid, such as N-protected lysine, in order to prepare a fixed N-protected polyamine having one or more primary amines. c) Depending on the circumstances, a process in which step (b) is repeated. d) Depending on the case, a step of reacting each primary amine in the fixed N-protected polyamine of step (b) or step c) with the linker portion. e) Preferably, in an automated flow peptide synthesis system (AFPS), a step of synthesizing the amino acid sequence bound to the fixed N-protected polyamine in step (b) or step (c), and to the linker portion in step (d), f) Depending on the case, a step to protect each N-terminus of each amino acid sequence, for example, a step to substitute each primary amine with an acyl or acetyl group. g) A step of removing each protecting group in order to form a product having at least one primary amine, h) Depending on the case, a step of reacting each primary amine with the linker portion, i) A step of reacting either the primary amine of the product of step (g) or the linker portion of the product of step (h) with a functional group. j) A step of cutting the product of step (i) from the solid support. A method for producing a product that includes [the specified element]. [Claim 32] The aforementioned product has the following structure: 【Chemistry 4】 [In the formula, n and m are independent integers of 1 or greater, where m + n is preferably 3 or greater.] Each L1 and L2 is independently the same or different linker. Each peptide comprises an amino acid sequence, preferably a therapeutic peptide. Each R is a functional group, X is a peptide containing two or more diamino acids. The method according to claim 31, having the following characteristics. [Claim 33] A peptide conjugate prepared by the method described in claim 31 or 32.

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