Antiviral polypeptide membrane fusion inhibitors, methods for preparing the same, and their use

Abstract

This invention provides polypeptide membrane fusion inhibitors for viruses of the Paramyxoviridae and / or Pneumoviridae families, as well as methods for preparing and using the same. [Solution] The present invention discloses compounds, medicinal salts thereof or derivatives thereof, pharmaceutical compositions and formulations containing the same, and the use of the same in the preparation of drugs for the prevention and / or treatment of diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

Description

[Technical Field]

[0001] "Cross-referencing of related applications" This disclosure claims priority to Chinese patent application 2025101651370, filed on 2025 / 2 / 14, and to Chinese patent application 2025113287192, filed on 2025 / 9 / 17. All contents of the aforementioned Chinese patent applications are incorporated herein by reference.

[0002] This disclosure relates to the biomedical technology field and to antiviral polypeptide membrane fusion inhibitors, methods for preparing the same, and their use. Specifically, it relates to certain polypeptides, in particular polypeptides of certain Paramyxoviridae and / or Pneumoviridae viruses, as well as the preparation of viral membrane fusion inhibitors and the use of said polypeptides in the preparation of drugs for the treatment or prevention of infections caused by Paramyxoviridae and / or Pneumoviridae viruses. [Background technology]

[0003] Viruses belonging to the families Paramyxoviridae and Pneumoviridae are two types of single-stranded negative-sense RNA viruses that pose a serious threat to human and animal health. Infections caused by these viruses are highly pathogenic and transmissible, and effective treatments are lacking, making them a significant global public health challenge.

[0004] Within the Paramyxoviridae family, the measles virus can cause acute respiratory infections and systemic complications in children, and there is a risk of localized outbreaks even in areas where vaccination is widespread. Non-suppurative parotid gland swelling caused by the mumps virus is often accompanied by serious complications such as orchitis and meningoencephalitis. On the other hand, emerging viruses such as the Nipah virus and Hendra virus have extremely high mortality rates and are transmissible from person to person, and are classified as biosafety level 4 (BSL-4) pathogens. Within the Pneumoviridae family, human respiratory syncytial virus (hRSV) is a major pathogen causing lower respiratory tract infections in infants and young children (bronchiolitis, pneumonia, etc.), resulting in millions of hospitalizations worldwide every year, and there is a shortage of specific therapeutic drugs. Human metapneumovirus (hMPV) causes acute respiratory illness in all age groups and poses a significant threat, especially to the elderly and immunocompromised individuals.

[0005] Currently, clinical interventions against the above-mentioned viruses have the following clear limitations: Existing vaccines are effective against only a few viruses, such as measles and mumps, and do not cover all circulating subtypes; antiviral drugs commonly used in clinical practice have limited therapeutic effects, strong toxic side effects (hemolytic anemia, teratogenicity, etc.), and are ineffective against many paramyxoviridae and pneumoviridae viruses; antibody drugs have some preventive effect against hRSV, but are expensive, have complex administration routes, and are prone to inducing drug-resistant mutations in the virus. Furthermore, the genomes of the two viruses are prone to mutation, which leads to a sustained decline in the effectiveness of existing interventions, further increasing the difficulty of treatment.

[0006] Against this backdrop, there is an urgent need for research and development of novel antiviral agents that possess broad-spectrum antiviral activity, have low toxic side effects, and are less likely to induce drug resistance. Antiviral polypeptides have advantages such as small molecular weight, clear mechanism of action, and good biocompatibility, and are gradually becoming an important direction in antiviral drug research and development. However, research on specific antiviral polypeptides against viruses of the Paramyxoviridae and Pneumoviridae families is still in the exploratory stage, and mature products that can be used for clinical treatment have not yet emerged. Therefore, there is an urgent need to develop novel antiviral polypeptides that can effectively inhibit infections from these two types of viruses in order to fill the gap in clinical treatment and address the public health threats posed by these viruses. [Overview of the project] [Problems that the invention aims to solve]

[0007] This disclosure provides membrane fusion inhibitory polypeptides to address the shortcomings of existing technologies.

[0008] Based on the inventor's prior research, this disclosure uses a polypeptide template such as sequence (1) (sequence number 103), (1)Z1-WJJLVOOSJJFDOOIJJVNOOIJJSLOOIJJSDOOLJJVNOOLJJTNOOITTI-Z3-Z2 In the template described above, Z1 is an amino terminus (NH2-) or a modification thereof, and Z2 is a carboxyl terminus (-COOH) or a modification thereof. Unless otherwise specified, an acetyl group (Ac-) is selected for Z1 and an amide group (-NH2) is selected for Z2, capping the amino and carboxyl terms of the polypeptide, respectively, to enhance the polypeptide's stability.

[0009] J is an acidic amino acid residue, which may be a glutamic acid residue or an aspartic acid residue, but is not limited to these. O is a basic amino acid residue, which may be a lysine residue or an arginine residue, but is not limited to these. A salt bridge is formed between J and O, exerting a universal stabilizing effect on the secondary structure of the polypeptide. The remaining positions are specific amino acid residues, which occupy corresponding positions in the RSV CHR natural sequence. These residues are mainly hydrophobic amino acid residues, forming a single hydrophobic surface that contacts and conforms to the drug target, resulting in a specific interaction. Z3 is a fatty acid modifying group, which includes a linker and a lipophilic group, the lipophilic group of which binds to the target cell of the drug.

[0010] Furthermore, specific polypeptides are designed using a training method for a polypeptide-receptor binding activity prediction model. [Means for solving the problem]

[0011] This disclosure solves the above technical problems with the following technical solutions.

[0012] Furthermore, polypeptide compounds of this disclosure are designed using a training method for a polypeptide-receptor binding activity predictive model.

[0013] In one embodiment, the present disclosure provides a compound, a medicinal salt thereof, or a derivative thereof, which comprises a polypeptide represented by one or more sequences selected from Formula I, Formula II, Formula III, Formula IV, SEQ ID NOs. 57-59, SEQ ID NOs. 82-88, and SEQ ID NOs. 97-102.

[0014] In another embodiment, the disclosure further provides the use of the compound, its medicinal salt or derivative thereof in the preparation of polypeptide membrane fusion inhibitors against Paramyxoviridae and / or Pneumoviridae viruses.

[0015] In another embodiment, the disclosure further provides isolated nucleic acid molecules encoding the compound, a medicinal salt thereof, or a derivative thereof.

[0016] In another embodiment, the disclosure further provides a recombinant vector containing the nucleic acid molecule.

[0017] In another embodiment, the disclosure further provides recombinant cells containing the nucleic acid molecule or recombinant vector.

[0018] In another embodiment, the present disclosure further provides a pharmaceutical composition comprising the compound, a medicinal salt thereof or a derivative thereof, the nucleic acid molecule, the recombinant vector, or the recombinant cell.

[0019] In another embodiment, the disclosure further provides a pharmaceutical product comprising the pharmaceutical composition.

[0020] In another embodiment, the present disclosure further provides a method for preparing the compound, its medicinal salt or derivative thereof, comprising using a carrier resin to sequentially couple the compound with a protected amino acid corresponding to a polypeptide amino acid sequence by a deprotection and coupling reaction to produce a salt-bridge-unmodified peptide.

[0021] In another aspect, the disclosure further provides the use of the compound, its medicinal salt or derivative thereof, the pharmaceutical composition or the pharmaceutical product in the preparation of drugs for the prevention and / or treatment of diseases caused by Paramyxoviridae and / or Pneumoviridae viruses.

[0022] In another embodiment, the Disclosure further provides a method for preventing and / or treating diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families, the method comprising administering an effective amount of the compound, its medicinal salt or derivative thereof, the pharmaceutical composition or the formulation product to a subject in need.

[0023] In another embodiment, the Disclosure further provides compounds, medicinal salts thereof, derivatives thereof, pharmaceutical compositions, or pharmaceutical products for preventing and / or treating diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

[0024] In another embodiment, the Disclosure further provides a method for inhibiting viruses of the Paramyxoviridae and / or Pneumoviridae families, the method comprising administering the compound, a medicinal salt thereof, or a derivative thereof, the pharmaceutical composition, or the formulation product to a sample.

[0025] Based on conformity with common sense in this field, each of the preferred conditions described above can be arbitrarily combined to obtain each of the preferred examples in this disclosure. [Effects of the Invention]

[0026] The beneficial effects obtained by this invention are as follows:

[0027] The polypeptides designed in this invention can effectively inhibit infection by respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus (hPIV). [Modes for carrying out the invention]

[0028] definition To facilitate understanding of this disclosure, we first define some terms. Furthermore, when listing possible ranges for a given value or parameter, please note that the purpose is to indicate that the intermediate values ​​and ranges of these cited values ​​also constitute part of this disclosure.

[0029] As used herein, the articles “one” and “one kind” ("a" and "an") refer to one or more grammatical objects of the article. By example, “one element” refers to one or more elements, such as multiple elements.

[0030] In this specification, the term "including" means and is used interchangeably with the phrase "including, but not limited to, ...".

[0031] In this specification, the term "or" means and is used interchangeably with the term "and / or" unless the context clearly indicates otherwise.

[0032] The abbreviations used in this disclosure have the following meanings: Ala (Alanine, A) Arg (Arginine, R) Arginine Asn(Asparagine, N)asparagine Asp(Asparticacid, D)Aspartic acid DCM (Dichloromethane) DMF (N,N-Dimethyl malonate) Dimethylformamide Env (Envelope glycoprotein) ESI-MS (Electronic spray ion mass spectrometry) Fmoc (Fluorenylmethoxycarbonyl) fluorenyl methoxycarbonyl group Gly (Glycine, G) Glycine Gln(Glutamine, Q) Glutamine Glu (Glutamic acid, E) Glutamic acid 6-HB (Six-Helix Bundle) HBTU 2-(1H-1-hydroxybenzotriazole)-1,1,3,3-tetramethylhexafluorophosphate His(Histidine, H) Histidine HoBt(1-Hydroxyl benzotiazole anhydrous)1-hydroxybenzotriazole NHR (N-terminal heptad repeat) CHR (C-terminal heptad repeat) HIV (Human immunodeficiency virus) HPLC (high-performance liquid chromatography) Ile (Isoleucine, I) isoleucine Leu (Leucine, L) Leucine Met (Methionine, M) Methionine Lysine (Lysine, K) Phe(Phenylalanine, F) Phenylalanine RSV (Respiratory Syncytial Virus) hPIV (human parainfluenza virus) hMPV (human metapneumovirus) Ser(Serine, S) TFA (Trifluoroacetic acid) Thr (Threonie, T) Threonine Tyr (Tyrosine, Y) tyrosine Val (Valine, V) Single-letter amino acid residues are represented as follows:

[0033] A: L-Alanine, C: L-Cysteine, D: L-Aspartic acid, E: L-Glutamic acid, F: L-Phenylalanine, G: L-Glycine, H: L-Histidine, I: L-Isoleucine, K: L-Lysine, L: L-Leucine, M: L-Methionine, N: L-Asparagine, P: L-Proline, Q: L-Glutamine, R: L-Arginine, S: L-Serine, T: L-Threonine, V: L-Valine, W: L-Tryptophan, Y: L-Tyrosine.

[0034] In one aspect, the present disclosure provides a compound comprising a polypeptide represented by any one or more sequences selected from Formula I, Formula II, Formula III, Formula IV, SEQ ID NO: 57-59, SEQ ID NO: 82-88 and SEQ ID NO: 97-102, a pharmaceutically acceptable salt thereof or a derivative thereof. Formula I (SEQ ID NO: 122) is X1-X2-X3-X4-X5-X6-X7-X8-X9-I-X 37 , 23 , 35 , 20 , 33 , 15 , 30 , 13 , 25 , 24 , 22 , 38 , 16 , 36 , 14 , 34 , 32 , 29 -E-X 13 -X 14 -X 15 -X 16 -IEE-X 20 -L-X 22 -X 23 -X 24 [[ID=2B]]-X 25 -ESD-X 29 -X 30 -L-X 32 -X 33 -X 34 -X 35 -X 36 -X 37 -X 38 where X1 is either D or absent, X2 is either E or absent, X3 is either W, F, D, or S or absent, X4 is either D or absent, X5 is either E, A, or K or absent, X6 is either F or S or absent, X7 is either D, N, or L or absent, X8 is either K, Q, or L or absent, X9 is either K or absent, X 11 is either E or N, X 13 is either V or E, X 14 is either N or K, X 15 is either K, R, or E16 is K or R, and X 20 is S, I, or L, and X 22 is K or R, and X 23 is K or R, and X 24 is I or H, and X 25 is E or N, and X 29 is K or R, and X 30 is K or R, and X 32 is either E or does not exist, X 33 is either E, V or does not exist, X 34 is either V, N, S or does not exist, X 35 is either N, K, D or does not exist, X 36 is either K or does not exist, X 37 is either K, A or does not exist, X 38 It is either L, A or does not exist. Equation II (sequence number 123) is, X'1-X'2-X'3-X'4-X'5-X'6-E-X'8-X'9-X' 10 -X' 11 -X' 12 -X' 13 -X' 14 -SQVNEKIN-X' 23 -SL-X' 26 -X' 27 -IR-X' 30 -X' 31 -X' 32 -X' 33 -X' 34 -KSDELL-X' 41 -X' 42 -X' 43 -X' 44 -X' 45 -X' 46 -X' 47 -X' 48 -X' 49 -X' 50 And, Here, X’1 is F, W, D, L or Y, X’2 is V or does not exist, X’3 is K or does not exist, X’4 is D, K or does not exist, X’5 is F, I or does not exist, X’6 is D or does not exist, X’8 is L or does not exist, X’9 is V or does not exist, X’ 10 is F or does not exist, X’ 11 is E or D, X’ 12 is A or I, X’ 13 is S or does not exist, X’ 14 is I or does not exist, X’ 23 is E or Q, X’ 26 is A or E, X’ 27 is F, E or K, X’ 30 is L or does not exist, X’ 31 is A or does not exist, X’ 32 is F or does not exist, X’ 33 is I or does not exist, X’ 34 is R or does not exist, X’ 41 is H or does not exist, X’ 42 is N or does not exist, X’ 43 is V or does not exist, X’ 44 is N or does not exist, X’ 45 is A or does not exist, X’ 46 is G or does not exist, X’ 47 is K, L or does not exist, X’ 48 is S or does not exist, X’ 49 is T or does not exist, X’ 50 is T or does not exist, Formula III (SEQ ID NO: 124) is X’’1 - X’’2 - X’’3 - X’’4 - D - X’’6 - X’’7 - IEEVN - X’’ 13 - X’’ 14 - IEESL - X’’ 20 - X’’ 21 - IEESD - X’’ 27 - X’’ 28 - L - X’’ 30 - X’’ 31-V-X’’ 33 -X’’ 34 and where X’’1 is W or F, X’’2 is D or does not exist, X’’3 is E or does not exist, X’’4 is F or does not exist, X’’6 is K or A, X’’7 is K or S, X’’ 13 is K or R, X’’ 14 is K or R, X’’ 20 is K or R, X’’ 21 is K or R, X’’ 27 is K or R, X’’ 28 is K or R, X’’ 30 is E or H, X’’ 31 is E or N, X’’ 33 is N or does not exist, X’’ 34 is A or does not exist, Formula IV (SEQ ID NO: 125) is WDEFDASISQ-X’’’ 11 -NEKINQSLEEIRKSDELLHN-X’’’ 32 -X’’’ 33 -X’’’ 34 -X’’’ 35 and where X’’’ 11 is V or does not exist, X’’’ 32 is V, N or does not exist, X’’’ 33 is N, A or does not exist, X’’’ 34 is A, L or does not exist, X’’’ 35 is L or does not exist.

[0035] In some preferred embodiments, the modified polypeptide has the structure of R1-XX-R2R3, where XX is the polypeptide as described above, R1 is an amino-terminal protecting group, R2 is absent or an optionally substituted linker, and R3 is a carboxyl-terminal protecting group.

[0036] In some embodiments, the compound, its medicinal salt, or its derivative comprises a polypeptide represented by any one or more sequences selected from SEQ ID NOs. 52 to 102.

[0037] In this disclosure, the polypeptide as defined in the preamble is a core sequence contained in the compound, its medicinal salt, or its derivative, the core sequence being a key portion that produces the inhibitory effect, and which can act on the NHR region of the F protein, thereby inhibiting the process by which the NHR and CHR form a hexahelix bundle in the F protein, a key step in viral infection, by forming an inactive hexahelix structure with the NHR or by disrupting the NHR structure, thereby preventing the fusion of the virus with the target cell. This inhibitory effect does not depend on modifications to both ends of the polypeptide, modifications of linkers or lipophilic compounds, etc.

[0038] In some embodiments, the compound, its medicinal salt, or its derivative comprises a polypeptide represented by one or more sequences selected from I-1, formula II-1, formula III-1, formula IV-1, and sequence numbers 106-121. Equation I-1 (sequence number 126) is, R1-X1-X2-X3-X4-X5-X6-X7-X8-X9-IX 11 -EX 13 -X 14 -X 15 -X 16 -IEE-X 20 -LX 22 -X 23 -X 24 -X 25 -ESD-X 29 -X 30 -LX 32 -X 33 -X 34 -X 35 -X 36 -X 37 -X 38 -R2-R3, Equation II-1 (sequence number 127) is, R1-X'1-X'2-X'3-X'4-X'5-X'6-E-X'8-X'9-I-X'11 -X' 12 -X' 13 -X' 14 -SQVNEKIN-X' 23 -SL-X' 26 -X' 27 -IR-X' 30 -X' 31 -X' 32 -X' 33 -X' 34 -KSDELL-X' 41 -X' 42 -X' 43 -X' 44 -X' 45 -X' 46 -X' 47 -X' 48 -X' 49 -X' 50 -R2-R3, Equation III-1 (Sequence No. 128) is, R1-X''1-X''2-X''3-X''4-D-X''6-X''7-IEEVN-X'' 13 -X'' 14 -IEESL-X'' 20 -X'' 21 -IEESD-X'' 27 -X'' 28 -L-X'' 30 -X'' 31 -V-X'' 33 -X'' 34 -R2-R3, Equation IV-1 (sequence number 129) is, R1-WDEFDASISQ-X''' 11 -NEKINQSLEEIRKSDELLHN-X''' 32 -X''' 33 -X''' 34 -X''' 35 -R2-R3, Sequence ID 106 is R1-ADAFRLEVNDASSKINESIEESLLSLEKLHNVNATA-R2-R3, Sequence ID 107 is R1-FDAFIQEINVNEDQSLEQSDELLLELHLLHSLLH-R2-R3, Sequence ID 108 is R1-FAEFNQKINQVNEKIEESLEEIRKSDEELHNVNATT-R2-R3, Sequence ID 109 is R1-SDEQVNEKINQSLAFIRRIRKLLHN-R2-R3, Sequence ID 110 is R1-ILELVNKKIEQSLKFIEKSDKLLEN-R2-R3, Sequence ID 111 is R1-SLEQVNKKINQSLKVNKKSDKLLEN-R2-R3, Sequence ID 112 is R1-FDEEVNKKIEQSLKINQSLEEIRKS-R2-R3, Sequence ID 113 is R1-SISQVNEKINEIQSLEEKSDKLLKS-R2-R3, Sequence ID 114 is R1-VNKKIEEEKQSLKKQSDKIEESDEN-R2-R3, Sequence ID 115 is R1-FDELVNKKIEKIEEVNKKSLKLLES-R2-R3, Sequence ID 116 is R1-FEVNRRRIEQSLEKSLESLEEEEHSDKKLHNELH-R2-R3, Sequence ID 117 is R1-SLEQVNKINKKIDKIEESLKKIEESDKKSLEVNKKL-R2-R3, Sequence ID 118 is R1-SLEQVNKINEKINKISQSLKKIEESDKKSDEVNAGL-R2-R3, Sequence ID 119 is R1-SLEQVNKKIEQSLESLKKSDKINQSLEEVNKSDELL-R2-R3, Sequence ID 120 is R1-SLEQVNEKINQSLAFIRKSDEINQSDEEVNKSDELL-R2-R3, Sequence ID 121 is R1-SDELVNKKIEFDKKINQSLKKIEESDKKKL-R2-R3, Here, R1 is an amino-terminal protecting group, preferably acetyl. R2 is either absent or optionally substituted, a linker, preferably -R4-R5(R6)-, where R4 is a polypeptide, and its amino acid sequence is preferably (EAAAK) m or (GSGSG) m And m is a natural number between 0 and 5, specifically, m may be any natural number among 0, 1, 2, 3, 4, and 5. R5 is lysine, cysteine, 2,3-diaminopropionic acid, ornithine, 2,4-diaminobutyric acid, or 2,7-diaminoheptanoic acid, preferably lysine. R6 is a lipophilic compound group modified from R5, preferably the lipophilic compound group is cholesterol, cholesterol succinic acid monoester, 2-cholesterol acetate, 2-cholesterol propionic acid, 3-cholesterol propionic acid, 2-cholesterol butyric acid, 2-cholesterol isobutyric acid, 3-cholesterol butyric acid, 3-cholesterol isobutyric acid, 4-cholesterol butyric acid, 2-cholesterol valeric acid, 2-cholesterol isovaleric acid, 3-cholesterol valeric acid, 5-cholesterol valeric acid, 2-cholesterol hexanoic acid, 6-cholesterol One or more selected from hexanoic acid, 2-cholesterol heptanoic acid, 7-cholesterol heptanoic acid, 2-cholesterol octanoic acid, 8-cholesterol octanoic acid, cholesterol bromoacetate, cholesterol chloride, palmitic acid, stearic acid, fatty acids containing 3 to 20 carbon atoms, aliphatic dicarboxylic acids containing 3 to 20 carbon atoms, and other groups that can act with cell membranes or viral membranes to enhance the interaction between the polypeptide and the cell membrane or viral membrane, more preferably cholesterol succinate monoester. R3 is a carboxyl-terminal protecting group, preferably -NH2.

[0039] In some embodiments, the compound, its medicinal salt, or its derivative comprises a polypeptide represented by any one or more sequences selected from numbers 1 to 51.

[0040] In some embodiments, the derivative is a solvate, a chelate, or a non-covalent complex.

[0041] In some embodiments, the medicinal salts include acetate, lactobionate, benzenesulfonate, laurate, benzoate, malate, bicarbonate, maleate, bisulfate, mandelate, bicarbonate, methanesulfonate, borate, brommethane, bromide, methyl nitrate, calcium edetate, methyl sulfate, d-camphorsulfonic acid, mucoate, carbonate, naphthalenesulfonate, chloride, nitrate, clavulanate, N-methylglucosamine, citrate, ammonium salt, dihydrochloride, oleate, ethylenediaminetetraacetate, oxalate, ethanedisulfonate, pamoate, This includes bishydroxynaphthalene, propionate lauryl sulfate, palmitate, ethanesulfonate, pantothenate, fumarate, phosphate / diphosphate, glucoheptone, polygalacturonate, glucosinolate, salicylate, glutamate, stearate, p-hydroxyphenylacetamidoenylarsonic acid, sulfate, hydroxybenzoate, basic acetate, succinate, hydrobromide, tannate, hydrochloride, tartrate, hydroxynaphthalene, 8-chlorotheophylline, iodide, toluenesulfonate, triethyliodine, lactic acid, valerate, etc. Depending on the application, the medicinal salts may be formed from cations such as sodium, potassium, and bismuth, or from bases such as ammonia, ethylenediamine, N-methylglutamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane, and hydroxytetramethylammonium. These salts can be prepared using standard methods, for example, by the reaction of a free acid with an organic base or an inorganic salt. When basic groups such as amino groups are present, acidic salts such as hydrochloride, hydrobromide, acetate, and pamoate can be used as dosage forms. When acidic or alcoholic groups are present, pharmaceutically acceptable esters such as acetate, maleate, and pivaloyloxymethyl chloride, as well as esters known from the literature to improve solubility and hydrolysis, can be used as sustained-release formulations and prodrug formulations.

[0042] In another embodiment, the Disclosure provides the use of compounds such as those described herein, their medicinal salts, or derivatives thereof in the preparation of polypeptide membrane fusion inhibitors against paramyxoviridae and / or Pneumoviridae viruses.

[0043] As described herein, “antiparamyxoviridae and / or pneumoviridae virus polypeptide membrane fusion inhibitors” are specially designed polypeptide drugs that can inhibit the fusion of the viral envelope to the host cell membrane during the paramyxoviridae and / or pneumoviridae viral infection process. By blocking this key step, the virus is unable to enter the host cell, thereby preventing viral replication and transmission.

[0044] In another aspect, the Disclosure provides isolated nucleic acid molecules encoding compounds such as those described in the Disclosure, medicinal salts thereof, or derivatives thereof.

[0045] In another aspect, the Disclosure provides a recombinant vector containing a nucleic acid molecule as described in the Disclosure.

[0046] In another aspect, the Disclosure provides recombinant cells containing nucleic acid molecules or recombinant vectors as described in the Disclosure.

[0047] In another embodiment, the Disclosure provides a pharmaceutical composition comprising a compound as described in the Disclosure, a medicinal salt thereof or a derivative thereof, a nucleic acid molecule as described in the Disclosure, a recombinant vector as described in the Disclosure, or a recombinant cell as described in the Disclosure, further optionally comprising a pharmaceutically acceptable carrier or excipient.

[0048] The pharmaceutical compositions of this disclosure may be solutions with or without a buffer, or compositions containing a pharmaceutically acceptable carrier. In this disclosure, the pharmaceutical compositions may be administered in solution. They may be administered in an unbuffered solution, for example, in saline or water. Alternatively, they may be administered in a suitable buffer solution. The buffer solution may include acetates, citrates, alcohol-soluble glutens, carbonates or phosphates, or any combination thereof. In preferred embodiments, the buffer solution is phosphate-buffered saline (PBS). The pharmaceutical composition may be made suitable for administration to a subject by adjusting the pH and volume molar osmotic concentration of the buffer solution.

[0049] In some embodiments, the buffer solution further comprises a reagent for controlling the osmolality of the solution so that the osmolality is maintained at a desired value, such as a physiological value in human plasma. Solutes that can be added to the buffer solution to control the osmolality include (but are not limited to) proteins, peptides, amino acids, non-metabolic polymers, vitamins, ions, sugars, metabolites, organic acids, lipids, or salts. In some embodiments, the reagent for controlling the osmolality of the solution is a salt. In some embodiments, the reagent for controlling the osmolality of the solution is sodium chloride or potassium chloride.

[0050] The pharmaceutical compositions of this disclosure can be administered in doses sufficient to inhibit RSV. Appropriate doses of the compounds of this disclosure used in mammals, particularly humans, are typically in the range of 0.1 mg / day to 100 mg / day, for example, in the range of 10 mg / day to 50 mg / day, or for example, in the range of 20 mg / day to 30 mg / day.

[0051] The pharmaceutical composition may be administered once daily, or it may be administered within a day in subdoses divided into two, three or more doses at appropriate intervals, or it may be administered by continuous infusion or delivery using a sustained-release formulation. In this case, the amount of compound contained in each subdose must be correspondingly reduced in order to achieve the total daily dose. The dose units may also be formulated for delivery over several days, for example, by using a common sustained-release formulation that provides sustained compound release over a period of several days. Sustained-release formulations are known in the art and are particularly useful for reagent delivery to specific sites, and thus can be used with the reagents of this disclosure. In this embodiment, the dose unit includes a number of corresponding daily doses.

[0052] In other embodiments, since a single dose of the pharmaceutical composition can be sustained for a long period, subsequent doses may be administered at intervals not exceeding 3, 4, or 5 days, or at intervals not exceeding 1, 2, 3, or 4 weeks. The treatment plan may be to administer once every 1 to 3 days for 4 to 7 consecutive days, and if the infection recurs, it may be administered for an additional 4 to 7 consecutive days, and the number of treatment cycles may be 1 to 7. In some embodiments of the present disclosure, a single dose of the pharmaceutical composition of the present disclosure is administered once a week. In other embodiments of the present disclosure, a single dose of the pharmaceutical composition of the present disclosure is administered once a month.

[0053] Those skilled in the art will understand that certain factors, including (but not limited to), the severity of the disease or condition, past treatments, the subject's overall health and / or age, and other pre-existing conditions, influence the dose and administration schedule required for effective treatment of the subject. Furthermore, treating a subject with a therapeutically effective dose of a composition may involve a single treatment or a series of treatments. As described in other parts of this specification, the effective dose and in vivo half-life of each compound covered by this disclosure can be estimated using general methods or based on in vivo studies using appropriate animal models.

[0054] Depending on whether local or systemic treatment is desired, and depending on the area of ​​treatment, the pharmaceutical compositions of this disclosure can be administered in many ways. Administration may be local (e.g., by transdermal patch), intrapulmonary (e.g., including inhalation or blowing of powder or aerosol, administration by nebulizer), intratracheal, intranasal, epidermal, and transdermal, oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, subdermal administration (e.g., via implantation device), intracranial administration (e.g., intracerebral, intrasacral), or intraventricular administration.

[0055] The pharmaceutical compositions of this disclosure (which may readily exist in unit dosage forms) can be prepared according to common techniques well known in the pharmaceutical industry. This type of technique includes the following steps: binding these active ingredients with the drug carrier or excipient. Generally, these pharmaceutical compositions are prepared by the following steps: uniformly and finely binding these active ingredients to a liquid carrier, a finely dispersed solid carrier, or both, and, if necessary, further forming a product.

[0056] In another embodiment, the Disclosure provides a pharmaceutical product comprising a pharmaceutical composition described herein.

[0057] Some of the pharmaceutical products of this disclosure further incorporate a carrier material into the pharmaceutical composition. The carrier material includes, but is not limited to, water-soluble carrier materials (polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), sparingly soluble carrier materials (ethylcellulose, cholesterol stearate, etc.), and enteric-coated carrier materials (cellulose acetate phthalate and carboxymethyl ethylcellulose, etc.). Water-soluble carrier materials are preferred. Multiple types of pharmaceutical products can be manufactured using these materials, and these pharmaceutical products include, but are not limited to, tablets, capsules, drops, aerosols, pills, powders, liquids, suspensions, emulsions, granules, liposomes, transdermal agents, lozenges, suppositories, lyophilized powder injections, etc. They may be general formulations, sustained-release formulations, controlled-release formulations, and various microparticle delivery systems. A wide variety of carriers known in the art can be used to manufacture unit-dose formulations into tablets. Examples of carriers include, for example, diluents and absorbents (starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, trehalose, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate, etc.), wetting agents and binders (water, glycerin, polyethylene glycol, ethanol, propanol, starch gel, dextrin, syrup, honey, glucose solution, gum arabic gel, gelatin gel, sodium carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinyl phosphate). Examples of additives include rolidone, disintegrants (dried starch, alginate, agar powder, fucoidan (brown algae starch), sodium bicarbonate, citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid ester, sodium dodecyl sulfonate, methylcellulose, ethylcellulose, etc.), disintegration inhibitors (sucrose, tristearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium salts, sodium dodecyl sulfate, etc.), and lubricants (talc, silica, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc.).Tablets can also be further manufactured into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, bilayer tablets, and multilayer tablets. A wide variety of carriers known in the art can be used to manufacture unit-dose dosage forms into pills. Examples of carriers include, for example, diluents and absorbents (glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, gelucire, kaolin, talc, etc.), binders (acalycium gum, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or wheat paste, etc.), and disintegrants (agar powder, dried starch, alginate, sodium dodecylsulfonate, methylcellulose, ethylcellulose, etc.). A wide variety of carriers known in the art can also be used to manufacture unit-dose dosage forms into suppositories. Examples of carriers include, for example, polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, etc. To prepare the unit dosage form into injectable formulations such as liquids, emulsions, lyophilized powder injections, and suspensions, all diluents commonly used in this field, such as water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxide isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters, can be used. Furthermore, to prepare an isotonic injection solution, an appropriate amount of sodium chloride, glucose, or glycerin can be added to the injectable formulation, and general solubilizers, buffers, pH adjusters, etc., can also be added. Additionally, if necessary, colorants, preservatives, fragrances, flavorings, sweeteners, or other materials may be added to the drug formulation. The above dosage forms can be administered by injection, including subcutaneous, intravenous, intramuscular, and intracavitary injections, as well as by intracavitary administration (rectal and vaginal administration), respiratory administration (nasal administration), and mucosal administration. Of the above administration routes, injection is preferred.

[0058] In some embodiments, the pharmaceutical compositions of the present disclosure can be manufactured into any of many possible formulations, which may be, for example, one of an oral preparation, an injection, an inhalation preparation, and a lyophilized preparation, but are preferred, such as an inhalation preparation, a lyophilized preparation, a subcutaneous injection, or an intramuscular injection.

[0059] In some embodiments, the method of administering the pharmaceutical product is selected from oral administration, injection administration, mucosal administration, transdermal administration, and spray administration, with administration by spraying into the lungs, subcutaneous injection, or intramuscular injection being preferred.

[0060] In another aspect, the present disclosure provides a method for preparing compounds, medicinal salts thereof, or derivatives thereof as described herein, the method being: The process includes the step of using a carrier resin to sequentially couple protected amino acids corresponding to the polypeptide amino acid sequence with a salt-bridge-unmodified peptide by deprotection and coupling reactions.

[0061] In some embodiments, the method further (1) Modification with a lipophilic compound, preferably modification with a cholesterol succinate monoester, (2) N-terminal acetylation capping step, and (3) comprising one or more C-terminal amidation capping steps.

[0062] In another aspect, the present disclosure provides the use of compounds such as those described in the present invention, medicinal salts thereof or derivatives thereof, and pharmaceutical compositions or pharmaceutical products described in the present disclosure, in the preparation of drugs for the prevention and / or treatment of diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

[0063] As used herein, “diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae” is intended to include any disease associated with infection by viruses of the Paramyxoviridae and / or Pneumoviridae.

[0064] In some embodiments, the Paramyxoviridae virus is selected from the genera Respiratory virus, Rubulavirus, Morbillivirus, and Henipavirus, and the Pneumoviridae virus is selected from the genera Metapneumovirus and Orthopneumovirus.

[0065] In some preferred embodiments, the virus of the genus Respiratory Virus is human parainfluenza virus (hPIV). The human parainfluenza virus includes hPIV-1, hPIV-2, hPIV-3, and hPIV-4 subtypes.

[0066] In some preferred embodiments, the virus of the genus Mumpsvirus is Mumpsvirus.

[0067] In some preferred embodiments, the virus of the genus Measles Virus is the measles virus.

[0068] In some preferred embodiments, the henipavirus virus is selected from nipahvirus and Hendravirus.

[0069] In some preferred embodiments, the virus of the genus Metapneumovirus is human metapneumovirus (hMPV).

[0070] In some preferred embodiments, the virus of the genus Orthopneumovirus is a respiratory syncytial virus (RSV).

[0071] In some specific embodiments, the human metapneumovirus is selected from human metapneumovirus types A1 (hMPV-A1), A2 (hMPV-A2), B1 (hMPV-B1), and B2 (hMPV-B2).

[0072] In some preferred embodiments, the respiratory syncytial virus is selected from RSV-A, RSV-B, and its variants.

[0073] In another embodiment, the present disclosure provides a method for preventing and / or treating diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families, the method comprising administering to a subject in need an effective amount of a compound described in the present disclosure, a medicinal salt thereof or a derivative thereof, a pharmaceutical composition as described in the present invention, or a pharmaceutical product as described in the present invention.

[0074] As used herein, “subject” is intended to include human or non-human animals, preferably mammals, such as mice. Most preferably, the subject or patient is human.

[0075] As used herein, “effective dose” means a dose sufficient to achieve treatment for a disease caused by a Paramyxoviridae and / or Pneumoviridae virus when administered to a patient for the treatment of such disease (e.g., to alleviate, improve, or maintain the symptoms of the pre-existing disease or one or more diseases). The “effective dose” may vary depending on the method of administration of the reagent, the disease and its severity, and the patient’s medical history, age, weight, family history, genetic makeup, stage of the pathological process induced by the Paramyxoviridae and / or Pneumoviridae viruses, the type of previous or concomitant treatment (if any), and other individual characteristics of the patient. The “effective dose” also includes doses that produce a desired local or systemic effect under a reasonable benefit / risk ratio suitable for any treatment. The compounds used in the methods of this disclosure can be administered in amounts sufficient to produce a reasonable benefit / risk ratio suitable for such treatment.

[0076] In another embodiment, the Disclosure provides compounds, medicinal salts thereof or derivatives thereof, pharmaceutical compositions thereof or pharmaceutical products thereof, for the prevention and / or treatment of diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

[0077] In another embodiment, the present disclosure provides a method for inhibiting viruses of the Paramyxoviridae and / or Pneumoviridae families, the method comprising administering to a sample a compound as described in the present invention, a medicinal salt thereof or a derivative thereof, a pharmaceutical composition as described in the present invention, or a pharmaceutical product as described in the present invention.

[0078] As used herein, the term “sample” includes similar bodily fluids, cells, or tissues separated from the body of a subject, and collections of bodily fluids, cells, or tissues present in the body of a subject. Examples of bodily fluids include blood, serum, serous exudate, plasma, cerebrospinal fluid, ocular fluid, lymph, urine, and saliva. Tissue samples may include samples from tissue, organs, or local areas. For example, a sample may originate from a specific organ, organ portion, or bodily fluids or cells within those organs. In some embodiments, “sample” refers to blood or plasma extracted from the subject.

[0079] As used herein, “for non-therapeutic purposes” means, for example, inhibiting the replication of Paramyxoviridae and / or Pneumoviridae viruses in a laboratory setting for scientific research purposes.

[0080] The following examples are for illustrative purposes only and do not limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are common methods familiar to those skilled in the art, and the raw materials used are all commercially available.

[0081] Example 1: Polypeptide Design [Table 1] TIFF2026137052000002.tif45163

[0082] [Table 2]

[0083] [Table 3]

[0084] [Table 4]

[0085] [Table 5] Here, Chol is cholesterol succinate monoester, and represents an ester formed by the linkage of the carboxyl group in the cholesterol succinate monoester, shown in the structure below, to the lysine amino group in the polypeptide.

[0086] [ka]

[0087] [Table 6]

[0088] [Table 7]

[0089] [Table 8]

[0090] [Table 9]

[0091] [Table 10]

[0092] [Table 11]

[0093] Example 2: Synthesis of unmodified peptides 1. Chemical reagents required for the preparation process The chemical reagents used, such as various Fmoc amino acids, N,N'-diisopropylcarbodiimide (DIC), 1-hydroxybenzotriazole (HOBt), N,N-dimethylformamide (DMF), piperidine (PIPE), ninhydrin, acetic anhydride (Ac2O), N,N-diisopropylethylamine (DIEA), trifluoroacetic acid (TFA), ethylenedithiol (EDT), thioanisole (TA), triisopropylsilane (TIPS), and phenol, were all purchased from major chemical reagent suppliers and were not further purified before use.

[0094] The protected amino acid raw materials used in the polypeptide synthesis process include Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Lys(Boc)-OH, Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Val-OH, and Fmoc-Trp(Boc)-OH. Here, the abbreviations have known definitions: Fmoc is a 9-fluorenyl methoxycarbonyl group, Dde is 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, Boc is a t-butoxycarbonyl group, tBu is a t-butyl group, OtBu is a t-butoxy group, Trt is a trityl group, and Pbf is (2,3-dihydro-2,2,4,6,7-pentamethylbenzofuran-5-yl)sulfonyl.

[0095] 2. Synthesis of peptide resins Using Rink Amide MBHA resin as a carrier resin, peptide resins were produced by sequentially coupling it with protective amino acids corresponding to the polypeptide amino acid sequence via de-Fmoc protection and coupling reactions.

[0096] 2.1 Incorporate the first protective amino acid into the main chain 0.3 mmol of the initial protective amino acid and 0.3 mmol of HOBt were taken, dissolved in an appropriate amount of DMF, and then 0.3 mmol of DIC was taken and slowly added to the DMF solution of the protective amino acid while shaking. The reaction was carried out at room temperature for 5 minutes to obtain an activated protective amino acid solution, which was then prepared for use.

[0097] 0.1 mmol of Rink Amide MBHA resin (0.35 mmol / g * 0.3 g) was taken and deprotected with a 25% PIPE / DMF solution (volume ratio) for 20 min (twice), then washed and filtered to obtain resin from which Fmoc had been removed.

[0098] The activated first protective amino acid solution was added to the resin from which Fmoc had been removed, and the coupling reaction was carried out for 60 minutes. After filtration and washing, a resin containing the first protective amino acid was obtained.

[0099] 2.2 Incorporating other protective amino acids into the main chain Using a method similar to the one described above for incorporating the first protective amino acid into the main chain, other protective amino acids corresponding to the polypeptide were sequentially incorporated to obtain a resin containing main chain amino acids.

[0100] Finally, the N-terminus was acetylated and capped using 0.3 mmol Ac2O + 0.6 mmol DIEA to complete the synthesis of the main chain.

[0101] After each of the above steps, the reaction was always controlled by detection using the Kaiser Test. If a certain amino acid condensation reaction was incomplete, the condensation was repeated once, and this was continued until the desired target peptide fragment was obtained.

[0102] 3. Preparation of the crude product The peptide resin described above was taken, a cleavage reagent (15 mL / g resin) was added, and after homogeneous mixing, the reaction was carried out at 30°C for 3 hours with shaking to cleave the target polypeptide from the resin and remove the side chain protecting groups. The filtrate of the reaction mixture was collected, the resin was washed three times with a small amount of TFA / DCM, the filtrates were combined, anhydrous ether was added to precipitate, and the mixture was centrifuged. The filtered cake was washed and precipitated twice with cold anhydrous ether, and all the liquid was removed to obtain a whitish powder, which is the unmodified peptide crude product.

[0103] Here, the composition and volume ratio of the cleavage reagent are trifluoroacetic acid:1,2-ethanedithiol:thioanisole:phenol:H2O:triisopropylsilane = 68.5:10:10:5:3.5:1.

[0104] 4. Preparation of refined products The unmodified peptide crude product described above was taken, dissolved in water / acetonitrile by stirring, and insoluble matter was removed by centrifugation to prepare it for use.

[0105] Purification was performed using reversed-phase high-performance liquid chromatography (HHL). The chromatography column model was Agela C18, with specifications of 10 μm, 100 Å, and 50 × 250 mm. Mobile phases were mobile phase A (aqueous solution of 0.05% TFA and 2% acetonitrile) and mobile phase B (aqueous solution of 90% acetonitrile). The mobile phase flow rate was 25 mL / min, the UV detection wavelength was 220 nm, and the elution method was gradient elution. The crude product solution was injected into the above chromatography column, the corresponding purified components were recovered, and the solvent was removed by direct lyophilization to obtain a purified trifluoroacetate polypeptide in a swollen state.

[0106] 5. Characterization of refined products The purified trifluoroacetate polypeptide was redissolved in water and acetonitrile, and a large amount of anion exchange resin (in acetate ion form) was added, followed by stirring for 3 hours. After filtration and washing of the ion exchange resin with a water / acetonitrile mixed solvent, the filtrates were combined and freeze-dried to obtain the expanded polypeptide acetate purified product (i.e., the unmodified peptides in Tables 1-1 to 1-5 and Table 1-11). The chemical structure of the unmodified peptides was characterized using a liquid chromatograph-mass spectrometer, and the purity, sequence structure, molecular weight, and purity of each unmodified peptide were detected using an analytical high-performance liquid chromatograph (chromatography column model: Agela C18, chromatography column specifications: 4.6 × 250 mm, flow rate: 1 mL / min).

[0107] conclusion Analytical HPLC verified that the purity of all synthesized unmodified peptides exceeded 95%, and mass spectrometry confirmed that they possessed the correct molecular weight.

[0108] Example 3: Synthesis of lipopeptides 1. Chemical reagents required for the preparation process The chemical reagents used were hydrazine hydrate and cholesterol succinate monoester. The protective amino acid raw material used in the polypeptide synthesis process is Fmoc-Lys(Dde)-OH.

[0109] 2. Synthesis of the complete sequences of the unmodified peptide and linker sequence. Here, the linker junction contains a peptide with the EAAAK sequence (SEQ ID NO: 104) and a peptide with the GSGSG sequence (SEQ ID NO: 105). Lipophilic compounds used for modification include cholesterol succinate monoester.

[0110] 2.1 Synthesis of the main chain (1) Synthesis of peptide resin: Using Rink Amide MBHA resin as a carrier resin, peptide resin was produced by sequentially coupling it with protective amino acids corresponding to the polypeptide amino acid sequence via de-Fmoc protection and coupling reactions.

[0111] (2) Incorporate the first protective amino acid into the main chain. 0.3 mmol of the initial protective amino acid and 0.3 mmol of HOBt were taken, dissolved in an appropriate amount of DMF, and then 0.3 mmol of DIC was taken and slowly added to the DMF solution of the protective amino acid while shaking. The reaction was carried out at room temperature for 5 minutes to obtain an activated protective amino acid solution, which was then prepared for use.

[0112] 0.1 mmol of Rink Amide MBHA resin (0.35 mmol / g * 0.3 g) was taken and deprotected with a 25% PIPE / DMF solution (volume ratio) for 20 min (twice), then washed and filtered to obtain resin from which Fmoc had been removed.

[0113] The activated first protective amino acid solution was added to the resin from which Fmoc had been removed, and the coupling reaction was carried out for 60 minutes. After filtration and washing, a resin containing the first protective amino acid was obtained.

[0114] (3) Incorporate other protective amino acids into the main chain Using a method similar to the one described above for incorporating the first protective amino acid into the main chain, other protective amino acids corresponding to the polypeptide were sequentially incorporated to obtain a resin containing the main chain amino acids. Finally, the synthesis of the main chain was completed by acetylation capping of the N-terminus with 0.3 mmol Ac2O + 0.6 mmol DIEA. After each of the above steps, the reaction was always controlled by detection using the Kaiser Test, and if an amino acid condensation reaction was incomplete, the condensation was repeated once until the desired target peptide fragment was obtained.

[0115] 2.2 Incorporation of side chains (1) The resin was treated with the smallest possible volume of 2% hydrazine hydrate / DMF solution (by volume) to remove the Dde protecting group from the C-terminal lysine side chain (10 min, twice), filtered and washed to obtain a resin from which Dde had been removed, and prepared for use.

[0116] (2) Lipophilic compound modification of C-terminal lysine of polypeptide Cholesterol succinate monoester modification of polypeptide C-terminal lysine: 0.3 mmol of cholesterol succinate monoester and 0.3 mmol of HOBt were taken, dissolved in an appropriate amount of DMF, and then 0.3 mmol of DIC was taken and slowly added to the solution containing cholesterol succinate monoester and HOBt, and the mixture was shaken for 5 minutes at room temperature. The prepared solution containing cholesterol succinate monoester, HOBt, and DIC was added to the resin from which Dde obtained in step (1) had been removed, and a coupling reaction was carried out for 60 minutes. The mixture was then filtered, washed, and dried to obtain a peptide resin.

[0117] Other chemical reagents, amino acid starting materials, and operating steps are the same as in Example 1.

[0118] conclusion Analysis of the HPLC results confirmed that the purity of all synthesized salt bridge polypeptides exceeded 98%, and mass spectrometry confirmed that the molecular weight of the polypeptides matched the theoretical molecular weight.

[0119] Example 4: Detection of polypeptide anti-RSV activity In the examples, the present disclosure uses the RSV-A long virus strain as the infecting virus, and measures the anti-RSV activity of the synthesized polypeptides by CPE detection, thereby identifying novel polypeptides that exhibited significantly superior activity compared to control polypeptide fusion inhibitors.

[0120] The inhibitory activity of the compounds listed in Tables 1-1 to 1-5, the positive polypeptides in Table 1-11, and the positive control group T-118 against RSV-A long virus strains was detected.

[0121] 1. Experimental materials Human laryngeal cancer cells (HEp-2) were obtained from the American Type Culture Collection (ATCC), with catalog number CCL-23. The cells were cultured in DMEM culture medium supplemented with 10% fetal bovine serum, 1% sodium pyruvate, 1% non-essential amino acids, 2 mM glutamine, 100 U / mL penicillin, and 100 μg / mL streptomycin. The RSV A long virus strain was also obtained from ATCC, with catalog number VR-26.

[0122] 2. Experimental Method In this study, the in vitro anti-RSV A long virus strain activity of the test sample was detected by applying plaque reduction experiments. Eight concentrations of the test sample and control compound were tested in double wells. HEp-2 cells were trypsin-digested and then diluted with experimental culture medium until the cell count reached 300,000 cells per mL. The diluted cells were added to a 96-well cell culture plate at a rate of 100 μL per well, resulting in 30,000 cells. The cells were cultured overnight in an incubator at 5% CO2 and 37°C. The following day, the test sample was serially diluted with experimental medium. The serially diluted sample and an equal volume of virus (100 PFU per well) were homogeneously mixed and incubated for 1 hour at 37°C and 5% CO2. Subsequently, the culture medium in the 96-well cell culture plate was discarded, and the sample-virus mixture (200 μL) was added to the 96-well cell culture plate and incubated for 2 hours at 37°C and 5% CO2. After 2 hours, the sample-virus mixture was discarded, and 200 μL of maintenance medium containing 0.8% CMC with the corresponding concentration of the test sample was added. Cell controls (cells, no compound treatment or viral infection) and viral controls (cells infected with virus, no compound treatment) were established. Cells were cultured for 1 day in an incubator at 5% CO2 and 37°C. One day after viral infection, the medium was discarded, the cells were fixed with 4% paraformaldehyde, and then permeabilized with 0.5% TritonX-100. After washing the plates with DPBS, RSV-specific antibody (RSV antibody, 1:3000 dilution) was added and incubated at 37°C for 1 hour. Then, secondary antibody (donkey anti-goat IgG, 1:500 dilution) was added and incubated at 37°C for 1 hour. The secondary antibody was discarded, TrueBlue solution was added, staining was performed for 10 minutes, the plates were thoroughly washed with running water, air-dried, and the number of spots per well was counted using a microplate imaging counter. The raw data is used to calculate the antiviral activity of the sample.

[0123] Cytotoxicity and antiviral experiments were conducted in parallel. HEp-2 cells were inoculated into microplates at a density of 30,000 cells per well and cultured overnight in an incubator at 5% CO2 and 37°C. The following day, diluted test samples were added. Cell controls (cells, no compound treatment) and culture medium controls (culture medium only, no cells or compound treatment) were established. The final concentration of DMSO in the culture medium was 0.5% in both cases. Cells were cultured for 1 day in an incubator at 5% CO2 and 37°C. Cell viability was detected using the CCK8 cell viability detection kit.

[0124] The raw data from the spot count and cell viability tests are used for testing the antiviral activity and cytotoxicity of the samples, respectively. The calculation formula is as follows:

[0125] % inhibition rate = 100 - (sample value - cell control mean) / (virus control mean - cell control mean) × 100 %Cell viability = (Sample value - Control average value of culture medium) / (Control average value of cells - Control average value of culture medium) × 100 Using GraphPad Prism (version 10), a nonlinear fitting analysis was performed on the inhibition rate of the sample, and the IC of the sample was determined. 50 and CC 50 The value was calculated.

[0126] I C 50 In in vitro antiviral activity experiments, IC 50 This represents an index of the inhibitory ability of the measured compound against infection of Hep-2 cells by the RSV A long virus strain. Specifically, it means that when the measured compound reaches a certain concentration, it can effectively inhibit infection of 50% of Hep-2 cells by the RSV A long virus strain, and this concentration is the IC50. 50 That is the case.

[0127] CC 50 In in vitro antiviral activity experiments, CC 50This is used to evaluate the degree of toxicity of the target compound to Hep-2 cells. More precisely, if the target compound is at a specific concentration, it can kill 50% of Hep-2 cells, and this concentration is measured in CC. 50 This is the definition. The fitting formula is log(inhibitor) vs. response -- Variable slope(four parameters).

[0128] The safety index SI is given by: SI = IC 50 / CC 50 That is the case.

[0129] The anti-RSV virus activity of polypeptide samples was detected using positive polypeptides 1-7 as control samples. The cell fusion inhibitory activity of the polypeptide samples and positive polypeptides is shown in Table 2.

[0130] [Table 12] TIFF2026137052000015.tif70163

[0131] conclusion 1. The anti-RSV virus activity of the designed salt-bridged unmodified peptide sample is very high. 2. The salt-bridged unmodified peptide samples prepared in this disclosure are highly safe, with safety index SI > 1 for all of them, and can be used as safe and effective anti-RSV virus surrogate drugs.

[0132] Example 5: Efficacy of polypeptides in studies of protective activity during viral challenge in mice 1. Experimental materials Female BALB / c mice were obtained from Shanghai Yishang Biotechnology Co., Ltd. Human laryngeal cancer cells (HEp-2) were obtained from the American Type Culture Collection (ATCC), with catalog number CCL-23. The cells were cultured in DMEM culture medium supplemented with 10% (v / v) fetal bovine serum, 1% penicillin, and streptomycin. The RSV A2 virus strain was obtained from ATCC, with catalog number VR-1540.

[0133] 2. Experimental Method Eight-week-old female BALB / c mice were divided into groups based on body weight (n=6). On Day 0, 1 × 10⁻¹⁰ mice were fed. 6 All animals underwent RSV nasal virus challenge with a viral challenge dose of pfu (20 μL) per animal. One hour after the viral challenge on Day 0, administration was started, and the polypeptide sample, positive control group, and solvent control group were administered tracheal spray once daily for 5 consecutive days (see Table 3 below for dosage and frequency). Two hours after the final dose on Day 4, lung tissue was collected from all animals. Left lung tissue was rapidly frozen in 10 times its volume of viral protection solution, stored at -70°C or below, and viral titer was detected using the plaque method.

[0134] For analysis, pulmonary RSV A2 virus was measured by plaque assay for viral load in tissue homogenates. Simply put, the day before sample detection, HEp-2 cells were inoculated into 12-well plates according to the standard procedure for culturing HEp-2 cells (inoculation density: 0.3 × 10⁶). 6Cells / mL (1000 μL cell suspension per well). On the incubation day, the supernatant of the tissue sample was taken after homogenization with a tissue homogenizer (maintaining a low temperature environment), centrifuged, and the supernatant was taken. The tissue homogenate sample was diluted using serum-free DMEM and inoculated into a 12-well plate with a final volume of 400 μL. After incubation for 2 hours, the homogenate supernatant was discarded, 1.5 mL of overlay (cell culture medium containing 1% soft agar) was added per well, and the cells were cultured in an incubator for approximately 72 hours. The overlay was discarded, the cells were fixed, blocked with blocking solution, stained with HRP-RSVG, color development was stopped with pure water, and the plates were air-dried. The colored spots on the well plates were counted using a fluorescent (enzyme-conjugated) immunospot reader, and the viral titer was expressed as plaque-forming units (PFU / g) per g of tissue. The viral titer was calculated as the arithmetic mean ± standard error of all animals in a group.

[0135] [Table 13]

[0136] Mice were subjected to a 1-hour viral challenge with RSV A2, followed by administration of compounds 13, 14, 17, 19, 20, 21, 26, 27, 28, and 40 once daily via airway spray for 5 consecutive days. Two hours after the final dose, lung tissue samples were collected from all groups, and viral plaques were detected. The results are shown in Table 3.

[0137] As can be seen from the results of drug efficacy experiments in mice, RSV compounds 13, 14, 17, 19, 20, 21, 26, 27, 28, and 40 can significantly reduce the viral titer in lung tissue.

[0138] Example 6: Efficacy of polypeptides in a study of protective activity during viral challenge in cotton rats. 1. Experimental materials Female cotton rats were obtained from Shanghai Yishang Biotechnology Co., Ltd. The RSV A2 virus strain was obtained from ATCC, catalog number VR-1540, the RT-qPCR primer + probe primer was obtained from Tsingke Biotech Co., Ltd., lot number TSP20241227-025-00187, and the RT-qPCR standard was obtained from Vazyme, lot number 20241008.

[0139] 2. Experimental Method Female Cotton rats (6-8 weeks old) were divided into groups according to body weight (n=5). The mice were manipulated according to the contents of Table 4. On Day 0, 1 × 10 5.5 All animals underwent RSV nasal virus challenge with a viral challenge dose of pfu (50 μL) per animal. One hour after viral challenge on Day 0, administration was started, and tracheal spray was administered once daily for 5 consecutive days to polypeptide samples, positive control groups, and solvent control groups. Two hours after the final dose on Day 4, lung tissue was collected from all animals. Left lung tissue (including some trachea) was rapidly frozen in 10 times its volume of viral protection solution, stored at -70°C or below, and viral titer was detected by RT-qPCR.

[0140] Steps in RT-qPCR: First, the samples were homogenized and lung tissue RNA was extracted. The supernatant of the tissue samples was taken after homogenization using a tissue homogenizer (maintaining a low-temperature environment). Then, approximately 100 μL of the homogenized supernatant was taken, and total RNA from the tissue was extracted using the VeZol reagent-chloroform-aqueous-magnetic bead method and dissolved in RNase-free water. The RT-qPCR reaction system was prepared as follows: A one-step RT-qPCR reaction system was prepared according to the RNA template, probe, primer pair, RT-qPCR buffer, and RNase-free water. Two double wells were set up for each sample, with 20 μL of the one-step RT-qPCR reaction system in each well. Simultaneously, a quantitative standard curve well (RSV A2 N gene full-length plasmid), a positive control well, and a negative control well were set up. Subsequently, the amplification program was set up according to reverse transcription, pre-denaturation, and cycle amplification. Amplification data was collected using a real-time fluorescence quantitative PCR instrument, and the mean value of the double-well CT values ​​was calculated. The CT values ​​of the sample detection results for each case were substituted into the quantitative standard curve to calculate the copy number. The formula for calculating the RSV viral load (copy number / g lung tissue) is: sample volume (copy number / g lung tissue) = copy number / relative tissue input (g).

[0141] See section 4 for experimental results.

[0142] [Table 14]

[0143] Cotton rats were subjected to a 1-hour viral challenge with RSV A2, followed by administration of compounds 13, 14, 17, 19, 20, 21, 26, 27, 28, and 40 once daily via airway spray for 5 consecutive days. Two hours after the last dose, lung tissue samples were collected from all groups, and viral titers were detected. The results are shown in Table 4.

[0144] As can be seen from the efficacy experiments of the compounds in cotton rats, RSV compounds 13, 14, 17, 19, 20, 21, 26, 27, 28, and 40 can significantly reduce the viral titer in lung tissue.

[0145] Example 7: Detection of antiviral activity Human metapneumovirus (hMPV) and human parainfluenza virus (hPIV) are two types of viruses that possess a typical class I fusion protein-F protein. The precursor F0 of the F protein is cleaved to form F1 and F2 subunits. Here, the HRN and HRC regions of the F1 subunit interact with each other during the structural change process to form a hexahelix bundle (6-HB), which promotes the fusion of the viral membrane with the host cell membrane. Using surface plasmon resonance (SPR) technology, hMPV-N46 (127-172) and hPIV-N51 (139-189), derived from the HRN region, were coupled onto a CM5 chip. By monitoring the changes in the SPR signal in real time as the compounds flowed along the chip surface, the dynamic processes of binding and dissociation were recorded. Furthermore, the binding rate constant (ka) and dissociation rate constant (kd) were obtained through dynamical analysis, and the equilibrium dissociation constant (KD = kd / ka) was calculated from these values.

[0146] 1. Experimental materials The items included a CM5 tip with catalog number 29149603, an amino group coupling kit with catalog number BR100050;HBS-EP+buffer(10×), sodium acetate (pH 4.0), and 10 mM glycine-hydrochloride buffer (pH 2.0), all purchased from GE Health.

[0147] 2. Experimental Method CM5 tip coupling: Channels 2 and 4 of the CM5 tip were activated for 7 minutes with a newly formulated 1:1 mixture of 50 mM Hydroxy succinimide (NHS) and 200 mM 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC). Then, hMPV-N46 or hPIV-N51 was diluted and dissolved in sodium acetate at pH 4.0 at a flow rate of 10 μl / min for 600 seconds until it reached 30 μM. The experimental buffer was HBS-EP + buffer (1×). Finally, the tip was blocked with 1 M ethanolamine. The instrument set temperature was 25°C.

[0148] Affinity detection: The buffer used in the experiment was HBS-EP+buffer(1×), and the multi-cycle power mode was selected. Channels 1 and 3 of the tip served as blank reference channels. The target polypeptide was flowed through channels 2 and 4 at concentrations of 0 μM, 0.04 μM, 0.12 μM, 0.37 μM, 1.11 μM, 3.33 μM, 10 μM, and 30 μM at a flow rate of 30 μl / min, binding for 180 s and dissociation for 120 s. Finally, the tip was regenerated by injecting with 10 mM glycine (pH 2.0). The instrument's set temperature was 25°C.

[0149] Data Analysis: The data was analyzed using Biacore T200 analysis software (Version 3.2.1), subtracting the reference channel and zero-concentration background signal, and analyzed using a 1:1 binding model.

[0150] The affinity detection results for polypeptide compounds are shown in Table 5.

[0151] [Table 15] TIFF2026137052000019.tif34163

[0152] Conclusion: Biological activity is a key quality characteristic that reflects the effectiveness of biological products, and special attention should be paid to the study and verification of biological activity. Receptor binding activity can be measured using surface plasmon resonance (SPR).

[0153] 1. The KD values ​​of the designed polypeptide compound and hMPV are in the range of μM to nM, which indicates moderate binding strength, and thus possesses a certain level of anti-hMPV viral activity.

[0154] 2. The KD values ​​of the designed polypeptide compound and hPIV are in the range of μM to nM, indicating moderate binding strength, which represents a certain level of anti-hPIV virus activity.

[0155] Example 8: Detection of anti-hMPV virus activity - plaque reduction experiment 1. Laboratory supplies African green monkey kidney cells (Vero) were obtained from ATCC, catalog number CCL-81. The cells were cultured in DMEM culture medium supplemented with 10% fetal bovine serum, 1% non-essential amino acids, 1% sodium pyruvate, 1% L-glutamine, 100 U / ml penicillin, and 100 μg / ml streptomycin.

[0156] The medium for hMPV plaque experiments is prepared by adding 1% non-essential amino acids, 100 U / ml penicillin, and 100 μg / ml streptomycin to Opti-MEM culture medium.

[0157] The human metapneumovirus (hMPV) A2 and B1 virus strains were constructed by WuXi AppTec Co., Ltd.

[0158] 2. Experimental Method In this study, we detected in vitro anti-hMPV A2 and B1 virus strain activity in test samples using plaque reduction experiments.

[0159] Eight concentrations were tested in double wells for both the test samples and the experimental system control. After trypsin digestion, Vero cells were diluted in a 2% serum-containing cell culture medium until the cell count reached 400,000 cells per mL. The diluted cells were added to a 96-well cell test plate at a concentration of 100 μL per well, totaling 40,000 cells. The cells were cultured overnight in a 5% CO2 incubator at 37°C.

[0160] The following day, the test samples were serially diluted in laboratory culture medium. After serial dilution, the sample was uniformly mixed with an equal volume of virus (approximately 200 PFU per well) and incubated at 33°C and 5% CO2 for 1 hour. Subsequently, the culture medium in the 96-well cell test plate was discarded, and the sample-virus mixture (100 μL) was added to the 96-well cell culture plate and incubated at 37°C and 5% CO2 for 2 hours. After 2 hours, the sample-virus mixture was discarded, and 200 μL of laboratory culture medium containing the corresponding concentration of the test sample was added. This culture medium contained 0.8% CMC and 5 μg / ml trypsin. The final concentration of DMSO in the culture medium was 0.5%. A control antibody was added directly to the laboratory culture medium containing 0.8% CMC and 5 μg / ml trypsin. Cell controls (cells, no compound treatment or viral infection) and viral controls (cell-infecting virus, no compound treatment) were established. The cells were cultured for one day in an incubator at 5% CO2 and 33°C.

[0161] Cells were fixed with 4% paraformaldehyde and then permeabilized with 0.5% TritonX-100. After washing the plates with DPBS, hMPV-specific antibody (1:2000 dilution) was added and incubated at 37°C for 1.5 hours. Then, secondary antibody (1:1000 dilution) was added and incubated at 37°C for 1 hour. The secondary antibody was discarded, TrueBlue solution was added and stained for approximately 10 minutes, the plates were thoroughly washed with running water, air-dried, and the number of spots per well was counted using a microplate imaging counter.

[0162] The raw data is used to test the antiviral activity of the sample. The calculation formula is as follows:

[0163] % Inhibition rate = 100 - (Sample value - Average cell control value) / (Average virus control value - Average cell control value) × 100 Using GraphPad Prism (version 10), non-linear fitting analysis was performed on the inhibition rate of the sample to calculate the EC 50 value of the sample. The fitting equation is log(inhibitor) vs. response--Variable slope (four parameters).

[0164]

Table 16

[0165] Example 9: Detection of anti-hPIV3 virus activity - Plaque reduction experiment 1. Experimental supplies African green monkey kidney cells (LLC-MK2) were obtained from ATCC, and the product number is CCL-7.1. The cells were cultured in DMEM medium supplemented with 10% fetal bovine serum, 1% non-essential amino acids, 1% sodium pyruvate, 1% L-glutamine, 100 U / ml penicillin, and 100 μg / ml streptomycin.

[0166] The medium for the HPIV-3 plaque experiment was prepared by adding 2% fetal bovine serum, 1% non-essential amino acids, 1% sodium pyruvate, 1% L-glutamine, 100 U / ml penicillin, and 100 μg / ml streptomycin to DMEM medium.

[0167] Human parainfluenza virus type 3 (hPIV3 C243) was obtained from ATCC, and the product number is VR-93.

[0168] 2. Experimental method This study applied a plaque reduction assay to detect the in vitro anti-hPIV3 C243 virus strain activity of the test samples, and Human Anti-HPIV3 Monoclonal Antibody, clone PIA174 was used as the experimental system control. For the test samples and the system control antibody, eight concentrations were tested in duplicate wells.

[0169] After LLC-MK2 cells were trypsinized, they were diluted with cell culture medium containing 2% serum to 400,000 cells per milliliter. The diluted cells were added to a 96-well cell test plate at 100 μL per well to achieve 40,000 cells per well. The cells were cultured overnight in an incubator at 5% CO2 and 37 °C.

[0170] The next day, an equal volume of virus (approximately 200 PFU per well) was uniformly mixed with the serially diluted samples and then incubated at 37 °C and 5% CO2 for 1 hour. Then, the culture medium in the 96-well cell test plate was discarded, and the sample-virus mixture (100 μL) was added to the 96-well cell culture plate and incubated at 37 °C and 5% CO2 for 2 hours. After 2 hours, the sample-virus mixture was discarded, and 200 μL of experimental medium containing the test sample at the corresponding concentration was added. The culture medium contained 0.8% CMC. The final concentration of DMSO in the culture medium was 0.5%. The control antibody was directly added to the experimental culture medium containing 0.8% CMC. Cell controls (no cells, compound treatment, or virus infection) and virus controls (virus-infected cells, no compound treatment) were set. The cells were cultured in a 5% CO2 and 37 °C incubator for 1 day. [[ID=​​Cells were fixed with 4% paraformaldehyde and then permeabilized with 0.5% TritonX-100. After washing the plates with DPBS, hPIV-specific antibody (1:2000 dilution) was added and incubated at 37°C for 1.5 hours. Subsequently, secondary antibody (1:500 dilution) was added and incubated at 37°C for 1 hour. The secondary antibody was discarded, TrueBlue solution was added, staining was performed for approximately 10 minutes, the plates were thoroughly washed with running water, air-dried, and the number of spots per well was counted using a microplate imaging counter.

[0172] The raw data is used to calculate the antiviral activity of the sample, and the calculation formula is as follows:

[0173] % inhibition rate = 100 - (sample value - cell control mean) / (virus control mean - cell control mean) × 100 Using GraphPad Prism (version 10), a nonlinear fitting analysis was performed on the inhibition rate of the sample to determine the EC of the sample. 50 The values ​​were calculated. The fitting formula is log(inhibitor) vs. response -- Variable slope(four parameters).

[0174] [Table 17] Although specific embodiments of the present invention have already been described in detail, those skilled in the art will understand that various modifications and substitutions are possible to those details in accordance with all the teachings already disclosed, and that any of these changes will fall within the scope of protection of the present invention. The entire scope of the present invention is defined by the appended claims and any equivalents thereof.

Claims

1. A polypeptide comprising one or more sequences selected from Formula I, Formula II, Formula III, Formula IV, Sequence IDs 57-59, Sequence IDs 82-88, and Sequence IDs 97-102, Equation I (Sequence ID 122) is, X １ -X ２ -X ３ -X ４ -X ５ -X ６ -X ７ -X ８ -X ９ -I-X １１ -E-X １３ -X １４ -X １５ -X １６ -IEE-X ２０ -L-X ２２ -X ２３ -X ２４ -X ２５ -ESD-X ２９ -X ３０ -L-X ３２ -X ３３ -X ３４ -X ３５ -X ３６ -X ３７ -X ３８ であり、 Here, X １ is either D or does not exist, X ２ is either E or does not exist, X ３ is either W, F, D, S or does not exist, X ４ is either D or does not exist, X ５ is either E, A, K or does not exist, X ６ is either F, S or does not exist, X ７ is either D, N, L or does not exist, X ８ is either K, Q, L or does not exist, X ９ is either K or does not exist, X １１ is E or N, and X １３ is V or E, and X １４ is N or K, and X １５ is K, R, or E, and X １６ is K or R, X ２０ is S, I, or L, and X ２２ is K or R, X ２３ is K or R, X ２４ is I or H, and X ２５ is E or N, and X ２９ is K or R, X ３０ is K or R, X ３２ is either E or does not exist, X ３３ is either E, V or does not exist, X ３４ is either V, N, S or does not exist, X ３５ is either N, K, D or does not exist, X ３６ is either K or does not exist, X ３７ is either K, A or does not exist, X ３８ It is either L, A, or does not exist. Equation II (Sequence ID 123) is, X' １ —X' ２ —X' ３ —X' ４ —X' ５ —X' ６ -E-X' ８ —X' ９ —X' １０ —X' １１ —X' １２ —X' １３ —X' １４ -SQVNEKIN-X' ２３ -SL-X' ２６ —X' ２７ -IR-X' ３０ —X' ３１ —X' ３２ —X' ３３ —X' ３４ -KSSDELL-X' ４１ —X' ４２ —X' ４３ —X' ４４ —X' ４５ —X' ４６ —X' ４７ —X' ４８ —X' ４９ —X' ５０ であり、 Here, X' １ is F, W, D, L or Y, and X' ２ is either V or does not exist, and X' ３ is either K or does not exist, and X' ４ is either D, K or does not exist, and X' ５ is either F, I or does not exist, and X' ６ is either D or does not exist, and X' ８ is either L or does not exist, and X' ９ is either V or does not exist, and X' １０ is either F or does not exist, and X' １１ is E or D, and X' １２ is A or I, and X' １３ is either S or does not exist, and X' １４ is either I or does not exist, and X' ２３ is E or Q, and X' ２６ is A or E, and X' ２７ is F, E or K, and X' ３０ is either L or does not exist, and X' ３１ is either A or does not exist, and X' ３２ is either F or does not exist, and X' ３３ is either I or does not exist, and X' ３４ is either R or does not exist, and X' ４１ is either H or does not exist, and X' ４２ is either N or does not exist, and X'<000011I>is either V or does not exist, and X' ４４ is either N or does not exist, and X' ４５ is either A or does not exist, and X' ４６ is either G or does not exist, and X' ４７ is either K, L or does not exist, and X' ４８ is either S or does not exist, and X' ４９ is either T or does not exist, and X' ５０ is either T or does not exist, Equation III (Sequence ID 124) is, X'' １ -X'' ２ -X'' ３ -X'' ４ -D-X'' ６ -X'' ７ -IEEEN-X'' １３ -X'' １４ -IEEEL-X'' ２０ -X'' ２１ -IEEED-X'' ２７ -X'' ２８ -L-X'' ３０ -X'' ３１ -V-X!!! ３３ -X'' ３４ And, Here, X'' １ is W or F, and X'' ２ is either D or does not exist, X'' ３ is either E or does not exist, X'' ４ is either F or does not exist, X'' ６ is K or A, and X'' ７ is K or S, and X'' １３ is K or R, and X'' １４ is K or R, and X'' ２０ is K or R, and X'' ２１ is K or R, and X'' ２７ is K or R, and X'' ２８ is K or R, and X'' ３０ is E or H, and X'' ３１ is E or N, and X'' ３３ is either N or does not exist, X'' ３４ is either A or does not exist. Formula IV (Sequence ID 125) is, WDEFDASISQ-X''' １１ -NEKINQSLEEIRKSDELLHN-X''' ３２ -X'' ３３ -X'' ３４ -X'' ３５ And, Here, X''' １１ is either V or does not exist, X''' ３２ is either V, N or does not exist, X''' ３３ is N, A, or does not exist, X''' ３４ is either A, L, or does not exist, X''' ３５ is either L or does not exist. A compound characterized by the above, its medicinal salt, or its derivative.

2. The polypeptide comprises one or more sequences selected from sequence numbers 52 to 102. The compound described in feature 1, its medicinal salt, or its derivative.

3. The structure of the aforementioned compound is R １ -XX-R ２ R ３ Here, XX is selected from the amino acid sequences shown in Formula I, Formula II, Formula III, Formula IV, SEQ ID NOs. 57-59, SEQ ID NOs. 82-88 and SEQ ID NOs. 97-102 of Claim 1, R １ R is an amino-terminal protecting group, ２ This is a linker that does not exist or has been arbitrarily replaced, R ３ This is a carboxyl-terminal protecting group, The compound described in feature 1, its medicinal salt, or its derivative.

4. The polypeptide comprises one or more sequences selected from formulas I-1, II-1, III-1, IV-1, and sequence numbers 106 to 121. Equation I-1 (Sequence ID 126) is, R １ -X １ -X ２ -X ３ -X ４ -X ５ -X ６ -X ７ -X ８ -X ９ -I-X １１ -E-X １３ -X １４ -X １５ -X １６ -IEE-X ２０ -L-X ２２ -X ２３ -X ２４ -X ２５ -ESD-X ２９ -X ３０ -L-X ３２ -X ３３ -X ３４ -X ３５ -X ３６ -X ３７ -X ３８ -R ２ -R ３ であり、 Equation II-1 (Sequence ID 127) is, R １ —X' １ —X' ２ —X' ３ —X' ４ —X' ５ —X' ６ -E-X' ８ —X' ９ —X' １０ —X' １１ —X' １２ —X' １３ —X' １４ -SQVNEKIN-X' ２３ -SL-X' ２６ —X' ２７ -IR-X' ３０ —X' ３１ —X' ３２ —X' ３３ —X' ３４ -KSSDELL-X' ４１ —X' ４２ —X' ４３ —X' ４４ —X' ４５ —X' ４６ —X' ４７ —X' ４８ —X' ４９ —X' ５０ -R ２ -R ３ であり、 Formula III-1 (Sequence ID 128) is, R １ -X'' １ -X'' ２ -X'' ３ -X'' ４ -D-X'' ６ -X'' ７ -IEEEN-X'' １３ -X'' １４ -IEEEL-X'' ２０ -X'' ２１ -IEEED-X'' ２７ -X'' ２８ -L-X'' ３０ -X'' ３１ -V-X!!! ３３ -X'' ３４ -R ２ -R ３ And, Formula IV-1 (Sequence ID 129) is, R １ -WDEFDASISQ-X''' １１ -NEKINQSLEEIRKSDELLHN-X''' ３２ -X'' ３３ -X'' ３４ -X'' ３５ -R ２ -R ３ That is, The compound according to feature 3, its medicinal salt, or its derivative.

5. The aforementioned R １ is acetyl, and / or the R ２ Ha-R ４ -R ５ (R ６ ) - and here, R ４ It is a peptide fragment, R ５ R is lysine, cysteine, 2,3-diaminopropionic acid, ornithine, 2,4-diaminobutanoic acid, or 2,7-diaminoheptanoic acid, ６ R ５ A lipophilic compound group linked to and / or the R ３ Ha-NH ２ And, Preferably, The aforementioned R ４ The amino acid sequence is (EAAAK) ｍ or (GSGSG) ｍ And m is a natural number selected from 0 to 5, and / or The lipophilic compound group is one or more selected from cholesterol, cholesterol succinate monoester, 2-cholesterol acetate, 2-cholesterol propionic acid, 3-cholesterol propionic acid, 2-cholesterol butyric acid, 2-cholesterol isobutyric acid, 3-cholesterol butyric acid, 3-cholesterol isobutyric acid, 4-cholesterol butyric acid, 2-cholesterol valeric acid, 2-cholesterol isovaleric acid, 3-cholesterol valeric acid, 5-cholesterol valeric acid, 2-cholesterol hexanoic acid, 6-cholesterol hexanoic acid, 2-cholesterol heptanoic acid, 7-cholesterol heptanoic acid, 2-cholesterol octanoic acid, 8-cholesterol octanoic acid, cholesterol bromoacetate, cholesterol chloride, palmitic acid, stearic acid, fatty acids containing 3 to 20 carbon atoms, and aliphatic dicarboxylic acids containing 3 to 20 carbon atoms, and the lipophilic compound group can strengthen the binding between the peptide fragment and the cell membrane or viral envelope by acting on the cell membrane or viral envelope. The compound according to feature 3 or 4, its medicinal salt, or its derivative.

6. The polypeptide comprises one or more sequences selected from numbers 1 to 51, The compound described in feature 1, its medicinal salt, or its derivative.

7. The derivative is a solvate, chelate, or non-covalent complex. A compound according to any one of claims 1 to 4, a medicinal salt thereof, or a derivative thereof.

8. Use of a compound according to any one of claims 1 to 4, a medicinal salt thereof, or a derivative thereof in the preparation of polypeptide membrane fusion inhibitors of antiparamyxoviridae and / or Pneumoviridae viruses.

9. A separated nucleic acid molecule characterized by encoding a compound according to any one of claims 1 to 5, a medicinal salt thereof, or a derivative thereof.

10. A recombinant vector characterized by containing the nucleic acid molecule described in claim 9.

11. Recombinant cells characterized by containing the nucleic acid molecule described in claim 9 or the recombinant vector described in claim 10.

12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7, a medicinal salt thereof or a derivative thereof, a nucleic acid molecule according to claim 9, a recombinant vector according to claim 10, or a recombinant cell according to claim 11, and optionally further comprising a pharmaceutically acceptable carrier or excipient.

13. The pharmaceutical composition comprises the one described in claim 12, Preferably, the dosage form of the pharmaceutical product includes any one from the group consisting of oral preparations, injectable preparations, inhaled preparations, and lyophilized preparations, with inhaled preparations, lyophilized preparations, subcutaneous injections, or intramuscular injections being preferred. A pharmaceutical product characterized by the following features.

14. The method of administration is selected from oral administration, injection, mucosal administration, transdermal administration, and spray administration, with lung spray administration, subcutaneous injection, or intramuscular injection being preferred. The pharmaceutical product according to feature 13.

15. A method for preparing a compound, a medicinal salt thereof, or a derivative thereof according to any one of claims 1 to 7, wherein the method is: The process includes the step of using a carrier resin to sequentially couple protected amino acids corresponding to the polypeptide amino acid sequence with a salt-bridge-unmodified peptide via deprotection and coupling reactions, Preferably, the method further includes: (1) Modification with a lipophilic compound, preferably a modification with cholesterol succinate monoester, (2) N-terminal acetylation capping step, (3) A C-terminal amidation capping step, comprising one or more of the above A method characterized by the following:

16. Use of a compound according to any one of claims 1 to 7, a medicinal salt thereof or a derivative thereof, a pharmaceutical composition according to claim 12, or a pharmaceutical product according to claim 13 or 14 in the preparation of a drug for the prevention and / or treatment of diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

17. The viruses of the Paramyxoviridae family are selected from the genera Respiratory Virus (Respirovirus), Mumps Virus (Rubulavirus), Measles Virus (Morbilivaryvirus), and Henipavirus (Henipavirus), and the viruses of the Pneumoviridae family are selected from the genera Metapneumovirus (Metapneumovirus) and Orthopneumovirus (Orthopneumovirus). Preferably, the virus of the respiratory virus genus is human parainfluenza virus, the virus of the mumps virus genus is mumps virus, the virus of the measles virus genus is measles virus, the virus of the henipavirus genus is selected from nipah virus and hendra virus, the virus of the metapneumovirus genus is human metapneumovirus, and the virus of the orthopneumovirus genus is respiratory syncytial virus (RSV). The use described in feature 16.

18. A method for preventing and / or treating diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families, characterized by administering an effective amount of a compound according to any one of claims 1 to 7, a medicinal salt thereof, or a derivative thereof, a pharmaceutical composition according to claim 12, or a pharmaceutical product according to claim 13 or 14 to a subject who requires it.

19. A compound according to any one of claims 1 to 7, a medicinal salt thereof or a derivative thereof, a pharmaceutical composition according to claim 12, or a pharmaceutical product according to claim 13 or 14, for preventing and / or treating diseases caused by viruses of the Paramyxoviridae and / or Pneumoviridae families.

20. A method for inhibiting viruses of the Paramyxoviridae and / or Pneumoviridae families, comprising administering to a sample a compound according to any one of claims 1 to 7, a medicinal salt thereof or a derivative thereof, a pharmaceutical composition according to claim 12, or a pharmaceutical product according to claim 13 or 14.

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