WO2014022947A1 - Short polypeptide inhibiting hiv and pharmacal use - Google Patents

Abstract

Disclosed in the present invention are a short polypeptide inhibiting HIV and the pharmacal use thereof. The polypeptide is the following a) or b) or c): a) the polypeptide HP25-N2 represented by SEQ ID NO: 14 in the sequence listing; b) a derived polypeptide with anti-HIV activity obtained by adding more than one amino acid residue to the amino terminal and/or the carboxyl terminal of the polypeptide represented by SEQ ID NO: 19 in the sequence listing, wherein the derived polypeptide consists of 21-30 amino acid residues; c) a derived polypeptide consisting of 21-30 amino acid residues obtained by adding more than one amino acid to any site in addition to the amino terminal and the carboxyl terminal of the polypeptide represented by SEQ ID NO: 19 in the sequence listing or substituting more than one amino acid therein, wherein the derived polypeptide has anti-HIV activity. Compared to other polypeptides, the polypeptide of the present invention is clearly short, but it has extremely strong antiviral activity, and has the advantages of ease of synthesis and low cost.

Description

 Short peptides and pharmaceutical use for inhibiting HIV

 The present invention relates to the field of biomedicine, and relates to a polypeptide for inhibiting HIV, a pharmaceutical composition thereof and use thereof. Background technique

 AIDS is mainly caused by human immunodeficiency virus type 1 (HIV-1). Since the discovery of AIDS in 1981, more than 60 million people worldwide have been infected with HIV, of which approximately 25 million have died. According to WHO statistics, about 2 million people are infected with HIV every year, which is a serious threat to human health. Vaccines are the best way to prevent AIDS, but effective AIDS vaccines are unlikely to have major breakthroughs in the near future. Therefore, the development of drugs that block different stages of replication of the virus is currently the key strategy for prevention and control of AIDS.

 The envelope glycoprotein (ENV) of HIV-1 mediates the process by which the virus enters the target cell. The protein is formed by the surface subunit gpl20 and the transmembrane subunit gp41 linked by non-covalent bonds. In its native state, ENV is a trimer in which gpl20 forms a globular complex and gp41 is inserted into the viral envelope (1). The main function of gpl20 is to bind to receptor CD4 and co-receptors (chemokine receptors CCR5 or CXCR4, etc.), while gp41 primarily mediates membrane fusion of viruses and cells. The study revealed that the extracellular domain of gp41 contains several important functional regions, including hydrophobic fusion peptides (FP), N-terminal helical repeats (NHR or HR1), and C-terminal helical repeats (CHR or HR2). As early as 1997, by analyzing the crystal structure of peptide complexes derived from NHR and CHR, it was found that the core structure of gp41 is a six-helix bundle (6-HB), and three NHRs consist of N- The helix interacts with the amino acid residues at the a and d positions to form a centrally located helical trimer with amino acid residues at the e and g positions exposed to the periphery of the centrosome and with a C-helix composed of three CHRs. The a and d positions interact (2-4). The C-helices are combined in an anti-parallel manner in the grooves formed by the three N-helices, respectively. Based on the three-dimensional structural information of gp41, the mechanism of HIV-1 membrane fusion was proposed: When gpl20 binds to the receptor on the target cell, a significant conformational change occurs, and the fusion peptide (FP) of gp41 is exposed and inserted into the target cell membrane, CHR and The reverse binding of NHR forms a stable 6-HB structure that fuses the viral membrane to the target cell membrane to mediate HIV entry into the target cell (5). The crystal structure reveals that NHR contains a distinct hydrophobic pocket and is considered a new target for anti-HIV drugs. For more than a decade, research into screening or designing HIV drugs for NHR pockets has been one of the hot spots.

Studies have shown that CHR contains three functional regions, namely the NHR hydrophobic pocket binding region (PBD: pocket-binding domain, aa628-635), the NHR binding region (NBD: aa628-666) and the lipid membrane binding region (LBD: Lipid-binding domain, aa666-673). The interaction between the NHR and the CHR helix plays a decisive role in the stability of the gp41 core structure and the viral infection activity. During the formation of the 6-HB core structure, the three amino acids of the PBD on the CHR (Trp628, Trp631 and Ile635) are inserted into the NHR hydrophobic pocket with high affinity, which is essential for stabilizing the six-helix bundle structure (6-7). However, the previously reported 6-HB structure CHR starts from Trp628, and the structure of its upstream sequence is not resolved (2, 4, 8), resulting in most studies based on the structure of SIV gp41 and presuming the upstream sequence of the CHR hydrophobic pocket binding region. ^{The 621} QIW NMT ⁶²⁷ may also exhibit a ct-helical structure (9). However, He, Y. et al found that peptide CP621-652 containing ⁶²¹ QIWNNMT ⁶²⁷ could form an extremely stable 6-HB (Tm = 82 °C) with NHR polypeptide, much higher than N36 and C34, which are considered to be gp41 core structure. Stability of the composite (Tm = 64 °C). At the same time, CP621-652 has significant antiviral activity (10). These results suggest that the ⁶²¹ QIW NMT ⁶²⁷ locus upstream of CHR has an important influence on the gp41-mediated membrane fusion process. To this end, He, Y. et al. proposed a modification of the concept of gp41 functional core structure. Recently, Chong, H. et al. performed structural analysis on 6-HB based on CP621-652 and found that the ⁶²¹ QIW NMT ⁶²⁷ sequence is not a helical structure (11). An exciting and important finding is that the amino acid residues Met626 and Thr627, which are adjacent to CHR, form a unique "hook"-like structure (designated MT hook) that stabilizes 6-HB by tightly "hooking" the hydrophobic pocket on the NHR. Core structure. The experimental results reveal that MT hooks play a crucial role in the infectivity of HIV. This finding lays the theoretical foundation for the in-depth development of MT-based HIV fusion inhibitors.

 The AIDS treatment drug T20 (Enfuvirtide, Fuzeon), which was approved by the US FDA "fast track" in 2003, is a 36 amino acid residue-derived polypeptide derived from gp41 CHR, which is mainly blocked by blocking the formation of 6-HB. Function (12). T20 is the first and currently the only HIV membrane fusion inhibitor for clinical treatment, but it has limited its widespread use due to drug resistance. Over the past decade or so, antiviral peptides targeting gp41 have been a hot spot in HIV drug research (13). The study found that the polypeptide C34 derived from gp41 CHR has higher antiviral activity than T20. Structurally both T20 and C34 contain an NHR binding region that interacts with the NHR sequence, but C34 contains a hydrophobic pocket binding region (PBD) and T20 lacks this motif. C34 is poorly soluble in water and difficult to develop into a drug, but because it represents the core sequence of CHR, it is widely used as a design template for novel antiviral peptides (13). T1249 is the second generation of T20, which has high HIV-inhibiting ability and is also very effective against T20-resistant strains, but clinical trials have been discontinued due to formulation problems (14). Research and development The third generation or the next generation of highly efficient fusion inhibitory peptides has important theoretical significance and application value. Based on the C34 sequence, the anti-HIV peptide Sifuvirtide was designed in China and Phase II clinical trials have been conducted (15). However, it is worrying that inhibitors based on C34 design are susceptible to the development of viral resistance (16-17).

 At the same time, it is interesting to note that most of the previously reported HIV fusion inhibitory polypeptides have longer amino acid sequences. For example, T20 and Sifuvirtide contain 36 amino acid residues, C34 contains 34 amino acid residues, and T1249 and T2635 have 39 amino acid residues. Undoubtedly, the length of the polypeptide is closely related to the difficulty and cost of its preparation. Recently, Japanese scholars reported a 29 amino acid polypeptide (SC29EK) with anti-HIV activity similar to C34, but further truncation (such as 22 amino acid SC22EK) greatly reduced the antiviral activity of the polypeptide and Stability of target binding (18-19). Recently, He Yuxian et al. designed a polypeptide containing 32 amino acids P32 based on an in-depth study of the structure and function of gp41 (Chinese invention patent application number:

201110112709. 7) , its anti-viral activity is significantly improved, especially showing strong advantages for T20 resistant viruses. references:

 1. Zhu, P., Liu, J., Bess, J., Jr., Chertova, E., Lifson, JD, Grise, H., Ofek, GA, Taylor, KA, and Roux, KH (2006) Distribution And three-dimensional structure of AIDS virus envelope spikes. Nature 441, 847-852

 2. Chan, D. C., Fass, D., Berger, J. M., and Kim, P. S. (1997) Core structure of gp41 from the HIV envelope glycoprotein. Cell 89, 263-273

3. Lu, M., Blacklow, SC, and Kim, PS (1995) A trimeric structural domain of The HIV-1 transmembrane glycoprotein. Nat Struct Biol 2, 1075-1082

4. Weissenhorn, W., Dessen, A., Harrison, S. C., Skehel, J. J., and Wiley, D. C. (1997) Atomic structure of the ectodomain from HIV-1 gp41. Nature 387, 426-430

 5. Eckert, D. M., and Kim, P. S. (2001) Mechanisms of viral membrane fusion and its inhibition. Annu Rev Biochem 70, 777-810

 6. Chan, D. C., Chutkowski, C. T., and Kim, P. S. (1998) Evidence that a prominent cavity in the coiled coil of HIV type 1 gp41 is an attractive drug target. Proc Natl Acad Sci U S A 95, 15613-15617

 7. Chan, D. C., and Kim, P. S. (1998) HIV entry and its inhibition. Cell 93, 681-684

8. Tan, K., Liu, J., Wang, J., Shen, S., and Lu, M. (1997) Atomic structure of a thermostable subdomain of HIV-1 gp41. Proc Natl Acad Sci USA 94, 12303 -12308

9. Caffrey, M., Cai, M., Kaufman, J., Stahl, SJ, Wingfield, PT, Covell, DG, Gronenborn, AM, and Clore, GM (1998) Three-dimensional solution structure of the 44 kDa ectodomain Of SIV gp41. EMBO J 17, 4572-4584

 10. He, Y., Cheng, J., Li, J., Qi, Z., Lu, H., Dong, M., Jiang, S., and Dai, Q. (2008) Identification of a critical motif For the human immunodeficiency virus type 1 (HIV-1) gp41 core structure: implications for designing novel anti-HIV fusion inhibitors. J Virol 82, 6349-6358

 11. Chong, H., Yao, X" Qiu, Z., Qin, B., Han, R., Waltersperger, S" Wang, M., Cui, S., and He, Y. (2012) Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652. J Biol Chem 287, 20281-20289

 12. Lalezari, JP, Henry, K., O'Heam, M., Montaner, JS, Piliero, PJ, Trottier, B. Walmsley, S., Cohen, C., Kuritzkes, DR, Eron, JJ, Jr. , Chung, J., DeMasi, R.,

Donatacci, L., Drobnes, C., Delehanty, J., and Salgo, M. (2003) Enfuvirtide, an HIV-1 fusion inhibitor, for drug-resistant HIV infection in North and South America. N Engl J eJ 348, 2175-2185

 13. Steffen, I., and Pohlmann, S. (2010) Peptide-based inhibitors of the HIV envelope protein and other class I viral fusion proteins. Curr Pharm Des 16, 1 143-1 158

14. Eggink, D., Baldwin, CE, Deng, Y., Langedijk, JP, Lu, M., Sanders, RW, and Berkhout, B. (2008) Selection of T 1249-resistant human immunodeficiency virus type 1 variants. J Virol 82, 6678-6688

 15. He, Y., Xiao, Y., Song, H., Liang, Q., Ju, D., Chen, X., Lu, H., Jing, W., Jiang, S., and Zhang, L. (2008) Design and evaluation of siflivirtide, a novel HIV-1 fusion inhibitor. J Biol Chem 283, 1 1 126-1 1 134

16. Shimura, K., Nameki, D., Kajiwara, K., Watanabe, K., Sakagami, Y., Oishi, S., Fujii, N., Matsuoka, M., Sarafianos, SG, and Kodama, EN (2010) Resistance profiles Of novel electrostatically constrained HIV-1 fusion inhibitors. J Biol Chem 285,

39471-39480

 17. Liu, Z., Shan, M., Li, L., Lu, L., Meng, S., Chen, C., He, Y., Jiang, S., and Zhang, L. (201 1 In vitro selection and characterization of HIV-1 variants with increased resistance to sifuvirtide, a novel HIV-1 fusion inhibitor. J Biol Chem 286, 3277-3287

18. Naito, T., Izumi, K., Kodama, E., Sakagami, Y., Kajiwara, K., Nishikawa, H., Watanabe, K., Sarafianos, SG, Oishi, S., Fujii, N. , and Matsuoka, M. (2009) SC29EK, a peptide fusion inhibitor with enhanced alpha-helicity, inhibits replication of human immunodeficiency virus type 1 mutants resistant to enfuvirtide. Antimicrob Agents Chemother 53, 1013-1018

 19. Nishikawa, H., Oishi, S., Fujita, M., Watanabe, K., Tokiwa, R., Ohno, H., Kodama, E., Izumi, K., Kajiwara, K., Naitoh, T ., Matsuoka, M., Otaka, A., and Fujii, N. (2008) Identification of minimal sequence for HIV-1 fusion inhibitors. Bioorg Med Chem 16, 9184-9187

Invention disclosure

 The technical problem to be solved by the present invention is to provide a polypeptide having a length of 21 to 30 amino acid residues having an anti-HIV activity, a pharmaceutically acceptable salt thereof, or a derivative thereof.

 The polypeptide, a pharmaceutically acceptable salt thereof, or a derivative thereof provided by the present invention, wherein the polypeptide is the following a) or b) or c):

 a) the polypeptide HP25-N2 shown in SEQ ID NO: 14 in the Sequence Listing;

 b) a derivative polypeptide having anti-HIV activity obtained by adding one or more amino acid residues at the amino terminus and/or the carboxy terminus of the polypeptide of SEQ ID NO: 19, the derivatized polypeptide consisting of 21-30 amino acid residues; a derivative polypeptide consisting of 21-30 amino acid residues obtained by adding or substituting one or more amino acids at any position including the amino terminus and the carboxy terminus of the polypeptide of SEQ ID NO: 19 in the sequence listing, the derivatized polypeptide having an antibiotic HIV activity.

 Wherein the sequence 14 in the sequence listing consists of 23 amino acid residues; the sequence 19 in the sequence listing consists of 18 amino acid residues, the amino acid sequence of which is the amino acid sequence shown in positions 3-20 of SEQ ID NO: 14 in the sequence listing. .

 In an embodiment of the present invention, the derivative polypeptide shown in b) is specifically any one of the following bl) - bl7):

 Bl ) amino acid sequence is the polypeptide ELT23 of sequence 18 in the sequence listing ;

 B2 ) the amino acid sequence is the polypeptide of sequence 9 in the sequence listing HP25 ;

 The amino acid sequence is the polypeptide of sequence 6 in the sequence listing , HP28 ;

 B4) the amino acid sequence is the polypeptide of sequence 10 in the sequence listing, HP24;

 B5 ) the amino acid sequence is the polypeptide HP30 of sequence 5 in the sequence listing;

 The amino acid sequence is the polypeptide of sequence 8 in the sequence listing , HP26 ;

 B7 ) the amino acid sequence is the polypeptide of sequence 7 in the sequence listing HP27 ;

The amino acid sequence is the polypeptide HP25-N3 of SEQ ID NO: 15 in the sequence listing; B9) the amino acid sequence is the polypeptide HP24-N2 of SEQ ID NO: 12 in the sequence listing;

 blO) the amino acid sequence is the polypeptide of sequence 17 in the sequence listing HP27-N3;

 Bl l ) amino acid sequence is the polypeptide of sequence 16 in the sequence listing HP26-N3;

 Bl2) the amino acid sequence is the polypeptide of sequence 13 in the sequence listing, HP24-N3;

 Bl3 ) the amino acid sequence is the polypeptide of sequence 11 in the sequence listing HP24-N1 ;

 Bl4) the amino acid sequence is the polypeptide of sequence 1 in the sequence listing MT-SC22EK;

 Bl5) the amino acid sequence is the polypeptide of sequence 2 in the sequence listing MT-SC21EK;

 Bl6) the amino acid sequence is the polypeptide of sequence 3 in the sequence listing MT-SC20EK;

 Bl7) The amino acid sequence is the polypeptide MT-SC19EK of sequence 4 in the Sequence Listing.

 The sequences of the 18 polypeptides listed in the specific embodiments of the present invention are shown in Table 1.

 Table 1. Polypeptide sequence

 ; No. Peptide name Amino acid length Polypeptide sequence SEQ ID NO :

 1 ; MT-SC22EK 24 MTWEEWDKKEEYTKKEELIKKS 1

2 MT-SC21 EK 23 MTWEEWDKKIEEYTKKIEELIKK 2 i 3 ; MT-SC20EK 22 MTWEEWDKKEEYTKKEELIK 3

; 4 ; MT-SC19EK 21 MTWEEWDKKIEEYTKKIEELI 4

5 HP30 30 WNEMTWEEWEKKIEEYTKKIEEILKKSEEQ 5

; 6 HP28 28 WNEMTWEEWEKKIEEYTKKIEEILKKSE 6

7 HP27 27 WNEMTWEEWEKKEEYTKKEEILKKS 7

8 HP26 26 WNEMTWEEWEKKIEEYTKKIEEILKK 8

; 9 HP25 25 WNEMTWEEWEKKEEYTKKEEILK 9

10 HP24 24 WNEMTWEEWEKKIEEYTKKIEEIL 10

11 HP24-N1 23 NEMTWEEWEKKIEEYTKKIEEIL 1 1

12 HP24-N2 22 EMTWEEWEKKEEYTKKEEIL 12

13 HP24-N3 21 MTWEEWEKKIEEYTKKIEEIL 13

14 HP25-N2 23 EMTWEEWEKKIEEYTKKIEEILK 14

15 HP25-N3 22 MTWEEWEKKEEYTKKEEILK 15

16 HP26-N3 23 MTWEEWEKKIEEYTKKIEEILKK 16

17 HP27-N3 24 MTWEEWEKKEEYTKKEEILKKS 17

18 ELT23 23 ELTWEEWEKKEEYTKKEEILK 18 wherein the abbreviations of amino acids have the meanings well-known in the art, for example: W is tryptophan, N is asparagine, E is glutamic acid, M is methionine, T is threonine, K Is lysine, I is isoleucine, Y is tyrosine, L is leucine, and S is serine.

 One or more amino acids of the polypeptide of the present invention may be substituted with a D-form amino acid, an artificially modified amino acid, a rare amino acid present in nature, etc., to enhance the bioavailability, stability and/or antiviral of the polypeptide. active. Wherein the D-type amino acid refers to an amino acid corresponding to the L-form amino acid constituting the protein; the artificially modified amino acid refers to a common L-type amino acid which constitutes a protein modified by methylation, phosphorylation or the like; the rare amino acid existing in nature includes a constituent protein Uncommon amino acids and amino acids that do not constitute proteins, such as 5-hydroxylysine, methylhistidine, gamma aminobutyric acid, homoserine, and the like.

 The above polypeptide derivative may be at least one of the following 1) to 5):

1) The amino terminus of any of the above polypeptides is linked to an amino terminal protecting group and/or a carboxy terminus of any of the above polypeptides a linker obtained by linking a carboxy terminal protecting group;

 2) a carboxy terminal of any of the above polypeptides linked to an oligopeptide or a lipophilic group or a linker obtained from cholesterol;

3) a linker obtained by linking an oligopeptide or a lipophilic group or cholesterol at an amino terminus of any of the above polypeptides;

4) a linker obtained by linking an oligopeptide or a lipophilic group or cholesterol to both the amino terminus and the carboxy terminus of any of the above polypeptides;

5) A modification obtained by modifying any of the above polypeptides with a protein, polyethylene glycol, or maleimide. Wherein, in the polypeptide derivative, the amino terminal protecting group may be an acetyl group, an amino group, a maleoyl group, a succinyl group, a tert-butoxycarbonyl group or a benzyloxy group or other hydrophobic group or a macromolecular carrier group. Any of the groups; the carboxy terminal protecting group can be any of NH ₂ , carboxyl, amide or t-butoxycarbonyl or other hydrophobic groups or macromolecular carrier groups. In an embodiment of the present invention, the polypeptide derivative is specifically a linker obtained by linking an amino group of the above-mentioned polypeptide to an acetyl group and a carboxy terminal of any one of the above polypeptides to an amide group.

 2) In the polypeptide derivative, the oligopeptide may be composed of 2-10 amino acid residues; the lipophilic group may have a fatty acid chain of 3 to 20 carbon atoms, such as a fatty acid having 8-16 carbon atoms. chain.

 Multimers formed from any of the above polypeptides and/or pharmaceutically acceptable salts thereof and/or derivatives thereof are also within the scope of the present invention. Wherein, a multimer refers to a polymer formed by two or more identical or different polypeptides linked together by amino acids (such as lysine and cysteine) or other molecules.

 Another technical problem to be solved by the present invention is to provide a composition.

 The present invention provides a composition comprising C1) and C2): C1) a polypeptide of any of the above, a derivative thereof, or a pharmaceutically acceptable salt thereof, and/or a multimer thereof; C2) pharmaceutically acceptable Carrier or accessory;

 The composition has at least one of the following D1) - D5):

 D1) anti-HIV;

 D2) treatment and / or prevention and / or adjuvant treatment of diseases caused by HIV infection;

 D3) inhibiting HIV for cell fusion;

 D4) inhibit HIV from invading cells;

 D5) inhibit HIV replication.

 Among them, the disease caused by HIV infection can be AIDS.

In practical applications, the polypeptide of the present invention, a pharmaceutically acceptable salt thereof, a derivative thereof, a multimer, and a composition can be directly administered to a patient as a medicament, or can be administered to a patient after mixing with a suitable carrier or excipient to achieve treatment. The purpose of HIV infection. The carrier materials herein include, but are not limited to, water-soluble carrier materials (such as polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), poorly soluble carrier materials (such as ethyl cellulose, cholesterol stearate, etc.), enteric carriers. Materials (such as cellulose acetate phthalate and carboxymethyl cellulose, etc.). Among them, preferred are water-soluble carrier materials. These materials can be used in a variety of dosage forms including, but not limited to, tablets, capsules, pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, Oral tablets, suppositories, freeze-dried powder injections, etc. It may be a general preparation, a sustained release preparation, a controlled release preparation, and various microparticle delivery systems. In order to form a unit dosage form into tablets, various carriers well known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, Sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate, etc.; wetting agents and binders, such as water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, Dextrin, syrup, honey, glucose solution, gum arabic, gelatin syrup, sodium carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants such as dried starch, alginic acid Salt, agar powder, brown algae starch, sodium hydrogencarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium dodecyl sulfate, methyl cellulose, ethyl cellulose, etc.; Inhibitors such as sucrose, glyceryl tristearate, cocoa butter, hydrogenated oils, etc.; absorption enhancers such as quaternary ammonium salts, sodium lauryl sulfate, etc.; lubricants such as talc, silica, corn Starch, stearate, boric acid, liquid paraffin, polyethylene glycol, and the like. Tablets may also be further formulated into coated tablets, such as sugar coated tablets, film coated tablets, enteric coated tablets, or bilayer tablets and multilayer tablets. In order to prepare a unit dosage form into a pill, various carriers well known in the art can be widely used. Examples of the carrier are, for example, a diluent and an absorbent such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gel _UC ire, kaolin, talc, etc.; binders such as gum arabic, gum tragacanth , gelatin, ethanol, honey, liquid sugar, rice paste or batter; etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecyl sulfate, methyl cellulose, ethyl cellulose, and the like. In order to prepare a unit dosage form as a suppository, various carriers well known in the art can be widely used. Examples of the carrier are, for example, polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides and the like. In order to prepare a unit dosage form into an injectable preparation such as a solution, an emulsion, a lyophilized powder, and a suspension, all diluents conventionally used in the art, for example, water, ethanol, polyethylene glycol, 1, may be used. 3-propanediol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid ester, and the like. Further, in order to prepare an isotonic injection, an appropriate amount of sodium chloride, glucose or glycerin may be added to the preparation for injection, and a conventional cosolvent, a buffer, a pH adjuster or the like may be added. In addition, coloring agents, preservatives, perfumes, flavoring agents, sweeteners or other materials may also be added to the pharmaceutical preparations as needed. The above dosage forms can be administered by injection, including subcutaneous injection, intravenous injection, intramuscular injection, and intraluminal injection; intraluminal administration, such as transrectal and vaginal; respiratory administration, such as transnasal; mucosal administration. The above administration route is preferably administered by injection.

 Usually, the pharmaceutical composition of the present invention contains 0.1 to 90% by weight of ^). Pharmaceutical compositions can be prepared according to methods known in the art. The dosage of the polypeptide of the present invention, its pharmaceutically acceptable salts, derivatives, multimers and compositions thereof depends on a number of factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, and weight of the patient or animal. And the individual reaction, the specific active ingredient used, the route of administration and the number of administrations, and the like. The above dosages may be administered in a single dosage form or divided into several, for example two, three or four dosage forms.

 The polypeptide of the present invention, a pharmaceutically acceptable salt thereof, a derivative thereof, a multimer and a composition thereof can be directly used for the treatment and prevention of HIV-infected persons, and can also be used in combination with one or more anti-HIV drugs to improve The purpose of the overall treatment effect. These anti-HIV drugs include, but are not limited to, reverse transcriptase inhibitors, protease inhibitors, invasion inhibitors, integration inhibitors, and maturation inhibitors. The above reverse transcriptase inhibitor may be AZT, 3TC, ddl, d4T, ddT, TDF, Abacavir Nevirapine Efavirenz and Delavirdine, etc., or several; the above protease inhibitors may be Saquinavir mesylate, Idinavir, Ritonavir ^

One or more of Amprenavir, Kaletra and Nelfmavir mesylate; the above-mentioned invasion inhibitor may be one or more of Maraviroc, TAK-779, T20, Τ2635, Sifuvirtide, VIRIP, etc.; The integration inhibitor can be Raltegravir or the like.

 A further aspect of the invention relates to a method of treating and/or preventing and/or adjuvant treatment of an envelope viral infection. The method comprises the steps of administering to the subject an effective amount of fl ) and / or / £2) and / or β):

 Π) any of the above polypeptides, pharmaceutically acceptable salts thereof, or derivatives thereof;

 F2) the above multimer;

 O) The above composition.

 Specifically, the envelope virus infection is a disease or AIDS caused by HIV infection.

 The specific therapeutically effective dose level for any particular patient will depend on a number of factors, including the disorder being treated and the severity of the disorder; the activity of the particular active ingredient employed; the particular composition employed. The age, weight, general health, sex and diet of the patient; the time of administration, the route of administration and the rate of excretion of the particular active ingredient employed; duration of treatment; in combination with or in combination with the particular active ingredient employed Drugs; and similar factors well known in the medical field. For example, it is the practice in the art to dose the active ingredient starting from a level lower than that required to achieve the desired therapeutic effect, gradually increasing the dosage until the desired effect is achieved. In general, the polypeptide of the present invention, a derivative thereof, or a pharmaceutically acceptable salt thereof, for use in mammals, particularly humans, may be administered at a dose of from 0.001 to 1000 mg/kg body weight per day, for example from 0.01 to 100 mg/kg. Body weight/day, for example between 0.01 and 10 mg/kg body weight/day.

 A further aspect of the invention also claims a nucleic acid molecule encoding any of the above polypeptides.

 Wherein, the nucleic acid molecule may be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule may also be R A , such as mR A or hnRNA.

 The invention also claims biomaterials of gl) or g2):

 Gl) an expression cassette, recombinant vector, recombinant virus or recombinant cell containing a nucleic acid molecule encoding any of the above polypeptides;

 G2) A recombinant expression vector, recombinant microorganism or recombinant cell expressing any of the above polypeptides.

 Wherein, expression is understood to include any step involved in the production of the polypeptide including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

Wherein the expression cassette is a single-stranded or double-stranded nucleic acid molecule comprising a nucleic acid molecule encoding any of the above polypeptides and all regulatory sequences necessary for its expression. The regulatory sequence, under compatible conditions, directs the coding sequence to express any of the above polypeptides in a suitable host cell. Such regulatory sequences include, but are not limited to, a leader sequence, a polyadenylation sequence, a propeptide sequence, a promoter, a signal sequence, and a transcription terminator. At a minimum, the regulatory sequences include promoters as well as termination signals for transcription and translation. In order to introduce a specific restriction enzyme site of the vector to ligate the regulatory sequence to the coding region of the nucleic acid sequence encoding the polypeptide, a linker-containing regulatory sequence can be provided. The control sequence may be a suitable promoter sequence, ie, a nucleic acid sequence that is recognized by the host cell expressing the nucleic acid sequence. The promoter sequence contains transcriptional regulatory sequences that mediate the expression of the polypeptide. The promoter may be any nucleic acid sequence that is transcriptionally active in the host cell of choice, including mutated, truncated and heterozygous promoters, which may be derived from extracellular or intracellular encoding homologous or heterologous to the host cell. The gene of the polypeptide. The control sequence may also be a suitable transcription termination sequence, a sequence that is recognized by the host cell to terminate transcription. The termination sequence is operably linked to the 3' end of the nucleic acid sequence encoding the polypeptide. Any terminator that can function in the host cell of choice can be used in the present invention. Regulation The sequence may also be a suitable leader sequence, an mR A untranslated region that is important for translation of the host cell. The leader sequence is operably linked to the 5' end of the nucleic acid sequence encoding the polypeptide. Any leader sequence that can function in the host cell of choice can be used in the present invention. The control sequence may also be a signal peptide coding region that encodes an amino acid sequence linked to the amino terminus of the polypeptide to direct the encoded polypeptide into the cell's secretory pathway. A signal peptide coding region which can direct the expressed polypeptide into the secretory pathway of the host cell used can be used in the present invention. The control sequence may also be a propeptide coding region that encodes an amino acid sequence at the amino terminus of the polypeptide. The resulting polypeptide is referred to as a zymogen or propolypeptide. A propolypeptide is generally inactive and can be converted to a mature active polypeptide by cleavage of the propeptide from the propolypeptide by catalytic or autocatalytic. When there is a signal peptide and a peptide domain at the amino terminus of the polypeptide, the peptide region is immediately adjacent to the amino terminus of the polypeptide, and the signal peptide region is adjacent to the amino terminus of the peptide region. It may also be desirable to add regulatory sequences that modulate the expression of the polypeptide depending on the growth of the host cell. Examples of regulatory systems are those that are capable of opening or shutting down gene expression in chemical or physical stimuli, including M reactions in the presence of regulatory compounds. Other examples of regulatory sequences are those that enable gene amplification. Sequences. In these examples, the nucleic acid sequence encoding the polypeptide should be operably linked to a regulatory sequence.

The invention also relates to a recombinant expression vector comprising a nucleic acid molecule, a promoter and a transcriptional and translational termination signal of the invention encoding any of the above polypeptides. When an expression vector is prepared, a nucleic acid molecule encoding any of the above polypeptides can be placed in a vector to be operably linked to appropriate expression control sequences. The recombinant expression vector can be any vector (e.g., a plasmid or virus) that facilitates recombinant DNA manipulation and expression of the nucleic acid sequence. The choice of vector will generally depend on the compatibility of the vector with the host cell into which it will be introduced. The vector can be a linear or closed loop plasmid. The vector may be an autonomously replicating vector (i.e., a complete structure that exists extrachromosomally and may be replicated independently of the chromosome), such as a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any mechanism to ensure self-replication. Alternatively, the vector is a vector that, when introduced into a host cell, will integrate into the genome and replicate along with the integrated chromosome. In addition, a single vector or plasmid may be employed, or two or more vectors or plasmids, or transposons, that will generally comprise all of the DNA that will be introduced into the genome of the host cell. The vector contains one or more selectable markers that facilitate selection of transformed cells. A selectable marker is a gene whose product confers resistance to a biocide or virus, resistance to heavy metals, or confers auxotrophs to prototrophy and the like. Examples of bacterial selection markers are the dal genes of Bacillus subtilis or Bacillus licheniformis, or the resistance markers of antibiotics such as ampicillin, kanamycin, chloramphenicol or tetracycline. The vector comprises elements that enable stable integration of the vector into the host cell genome, or that ensure that the vector autonomously replicates in the cell independently of the cell genome. In the case of autonomous replication, the vector may also contain an origin of replication enabling the vector to replicate autonomously in the host cell of interest. The origin of replication may be mutated such that it becomes temperature sensitive in the host cell (see, e.g., ffihrlich, 1978, Proc. Natl. Acad. Sci. USA 75: 1433). More than one copy of a nucleic acid molecule of the invention encoding any of the above polypeptides can be inserted into a host cell to increase the yield of the gene product. An increase in the copy number of the nucleic acid molecule can be accomplished by inserting at least one additional copy of the nucleic acid molecule into the host cell genome, or by inserting an amplifiable selectable marker with the nucleic acid molecule, by culturing the cell in the presence of a suitable selection reagent A cell containing an amplified copy of the selectable marker gene, thereby containing the additional copy nucleic acid molecule, is selected. The procedures for ligating the above elements to construct the recombinant expression vectors of the present invention are well known to those skilled in the art (see, for example, Sambrook et al., Molecular Cloning Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor , New York, 1989). The term "operably linked" is defined herein as a conformation wherein the regulatory sequences are located at appropriate positions relative to the coding sequence of the DNA sequence such that the regulatory sequences direct expression of the polypeptide.

 A further aspect of the invention relates to a recombinant cell comprising a nucleic acid molecule encoding any of the above polypeptides. The recombinant cell can be a prokaryotic cell or a eukaryotic cell, such as a bacterium (such as an E. coli cell) or a yeast cell.

 Still another aspect of the present invention also claims a method of preparing the following products.

 The preparation method of the product, including the steps of el) or e2):

 El) a step of chemically synthesizing any of the above polypeptides;

 E2) a step of biologically expressing any of the above polypeptides;

The product is fl), Ώ) or β) _:

 Fl) any of the above polypeptides, pharmaceutically acceptable salts thereof, or derivatives thereof;

 F2) the above multimer;

 O) The above composition.

 The biological expression of the polypeptide may specifically be the expression of the polypeptide in a biological cell. The biological cell can be a non-human animal cell, a microbial cell or a plant cell.

 The chemical synthesis may specifically be solid phase synthesis or liquid phase synthesis.

DRAWINGS

 Figure 1 shows the anti-HIV activity of the short peptide HP25-N2 and its comparison with the control polypeptide.

 Figure 2 shows the circular dichroism analysis of the interaction between the anti-HIV short peptide and the NHR polypeptide N36 based on SC22EK. Figure 3 is a circular dichroism analysis of the interaction between the anti-HIV short peptide and the NHR polypeptide N36 based on the P32 truncation design. The best way to implement the invention

 The embodiments of the present invention are described in detail below with reference to the accompanying drawings. In the examples, the specific conditions are not specified, and are carried out according to the general conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not specified by the manufacturer, and are all conventional products that are commercially available. The amino acids of all of the polypeptides in the following examples are L-form amino acids.

 Example 1. Preparation of polypeptide

All of the polypeptides used in the examples were synthesized by standard solid phase peptide synthesis (Fmoc/tBu strategy) and manually synthesized from the carboxy terminus to the amino terminus. All polypeptide sequences are amidated at the C-terminus, N-terminal acetylation, according to conventional polypeptide synthesis (as known to those skilled in the art, these modifications have no fundamental effect on polypeptide activity). N-fluorenylmethoxycarbonyl (Fmoc) protected amino acid, Rink resin (substitution constant 0.44 mmol/g) as solid phase carrier, and 25% (by volume) piperidine in DMF solution to remove amino protecting group Fmoc, each removal step needs to be performed twice, for 8 min and 10 min respectively. The condensation method selected for the peptide reaction was DIPC/HOBt method and PyBOP method, and the amino acid and the activating reagent were all used in three-fold equivalents, and the reaction time was 1 hour. The reaction progress was monitored by ninhydrin qualitative color development (Kaiser method). If the condensation reaction of an amino acid is incomplete, the reaction time is appropriately extended or the condensation is repeated once until the desired target peptide is obtained. Using a cleavage reagent (trifluoroacetic acid: 1,2-ethanedithiol: thioanisole: phenol: 3⁄40: triisopropylsilane = 68.5:10:10:5:3.5:1, v/v) The polypeptide was cleaved from the resin and the side chain protecting groups were removed (30 C cut for 4 hours). The filtrate was added to a large amount of cold anhydrous diethyl ether to precipitate a polypeptide and centrifuged. After washing several times with diethyl ether, it was dried to obtain a crude polypeptide.

 The prepared polypeptide was purified using a HP1100 (Agilent, USA) reversed-phase high performance liquid chromatograph. Column type: Cosmosil 5C4-AR C10 X 250 mm). Chromatographic operating conditions: Linear gradient elution, the eluent consists of mobile phase A and mobile phase B. The mobile phase A was an aqueous solution of trifluoroacetic acid and acetonitrile, the volume fraction of trifluoroacetic acid was 0.05%, and the volume percent concentration of acetonitrile was 2%. Mobile phase B is 90% (by volume) aqueous acetonitrile. The linear gradient elutes from 20% B to 40% B for 20 minutes, the elution flow rate is 25 ml per minute, and the UV detection wavelength is 220 nm. After lyophilization, the pure peptide was obtained in a fluffy state. The chemical structure was characterized by MALDI-TOF mass spectrometry and the purity was determined by analytical high performance liquid chromatography (flow rate: 1 ml per minute). Among them, Analytical High Performance Liquid Chromatograph Model: Shimadzu CBM-10A VP PULS, the type of column used: Agela4.6*250mm C18. Chromatographic operating conditions: Linear gradient elution, the eluent consists of mobile phase A and mobile phase B. The mobile phase A was an aqueous solution of trifluoroacetic acid and acetonitrile, the volume fraction of trifluoroacetic acid was 0.05%, and the volume percent concentration of acetonitrile was 2%. The mobile phase B is an aqueous solution of trifluoroacetic acid and acetonitrile, the volume fraction of trifluoroacetic acid is 0.05%, and the volume fraction of acetonitrile is 90%. The linear gradient elutes from 25% B to 45% B for 20 minutes. The obtained pure polypeptide was characterized by analytical RP-HPLC to indicate that the purity was more than 95%. The molecular weight and purity of the polypeptide of the present invention obtained according to the above polypeptide synthesis method are shown in Table 2.

 Table 2. Molecular weight and purity of the polypeptide

 No. Peptide name Theoretical molecular weight Actual molecular weight Purity (%)

1 ; MT-SC22EK ; 31 26.65 31 27.93 98

2 ; MT-SC21 EK ; 3039.57 3040.1 99

3 ; MT-SC20EK ; 291 1 .39 291 2.2 95

4 ; MT-SC1 9EK ; 2783.22 2784.2 97

5 HP30 3956.47 3956.65 98

6 HP28 3700.23 3701 .63 98

7 HP27 3570.1 1 3571 .1 7 96

8 HP26 3484.04 3484.45 95

9 HP25 3355.86 3356.08 99

10 HP24 3227.69 3228.37 97

11 HP24-N1 3040.47 3041 .55 95

12 HP24-N2 2926.36 2927.23 98

13 HP24-N3 2797.24 2797.39 99

14 HP25-N2 3054.54 3055.78 97

15 HP25-N3 2925.42 2925.99 98

16 HP26-N3 3053.6 3054.03 97

17 HP27-N3 3140.67 3141 .91 99

18 ELT23 3036.5 3037.79 97 Example 2. Discovery and identification of highly active anti-HIV short peptides

 2.1 Experimental materials and methods

 2.1.1 Inhibition of HIV-mediated cell fusion by peptides

HIV-mediated inhibition of cell fusion assays (Chong H, Yao X, Qiu Z, Qin B, Han R, Waltersperger S, Wang M, Cui S, He Y. Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight Of HIV-1 Fusion Inhibitor CP621-652. J Biol Chem. 2012, 287(24): 20281-20289). The target cells are TZM-bl cells, provided by the National Institutes of Health (NIH) AIDS Reagents and References Project (catalog number 8129), which express CD4 and CCR5 and CXCR4 on the surface, and fluoresceinase reporter gene in the cell. But does not contain the promoter of this gene. The effector cells are HL2/3 cells, which are provided by the NIH AIDS Reagents and References Program (catalog number 1294), which express HIV Env on the surface to mediate fusion of target cells, and also contain fluorescein in the cells. The promoter of the enzyme reporter gene. Both cells were adherent cells cultured in DMEM cell culture medium containing ammonia/streptomycin double antibody and 10% fetal bovine serum. TZM-bl was first added to a 96-well cell culture plate (lxlO ⁴ cells/well), and cultured overnight at 37 ° C under 5% CO ₂ . The test polypeptide was diluted 3-fold in DMEM cell culture medium, mixed with HL2/3 effector cells, and then added to TZM-bl target cells, and culture was continued for 6 hours to fully fuse. The activity of the Promega luciferase reporter gene kit (relative fluorescence unit, RLU) was then used. Calculate each concentration of sample to suppress the use of GraphPad Pri sm

Software 2. 01 software calculates the half effective inhibitor amount (IC ₅ . value).

 2.1.2 polypeptide inhibition HIV invasion cells

The HIV recombinant pseudovirus system was used to evaluate the inhibitory effect of the polypeptide on the target cells (Yao X, Chong H, Zhang C, Waltersperger S, Wang M, Cui S, He Y. Broad antiviral activity and crystal structure of HIV-1 fusion inhibitor Sifuvirtide. J Biol Chem. 2012, 287: 6788-6796. ). The basic steps include (1) preparation of HIV pseudovirus: plasmid pcDNA3.1-ENV expressing the HIV ENV protein (recombinant expression plasmid obtained by inserting the ENV encoding gene into vector pcDNA3.1 (-)) and HIV backbone plasmid pSG3 ^AENV ( Express all proteins except the ENV in the HIV genome, provided by the NIH AIDS Reagents and References Project, catalog number 11051), transfect 293T cells at a mass ratio of 1:2, and simultaneously transfect only the same amount. Control of ^{pSG3 AENV} . After incubating for 6 hours at 37 ° C in a 5% CO ₂ incubator, the solution was changed, and then incubation was continued for 48 hours to secrete the pseudovirus into the supernatant. Use a pipette to aspirate as much as possible of the supernatant from the cell culture flask or cell culture plate, filter through a 0.45 μηι filter or centrifuge at 1000 g for 10 minutes, and add the supernatant to the fetal bovine serum (FBS) to a final concentration of 20%. Transfer to a polypropylene tube for storage at 80 ° C or directly for virus titration. (2) Titration of HIV pseudovirus: The virus was diluted 5-fold in a 96-well plate, and 8 gradients of 4 replicate wells were set, and the final volume was ΙΟΟμΙ. TZM-bl cells were trypsinized and counted. The cells were diluted to 1× ^{10 5} cells/ml in DMEM complete medium, and ΙΟΟμΙ cells (containing 15 g/ml DEAE-dextran) were added to each well at 37 ° C, 5%. C0 _{2 was} cultured for 48 hours. Then, the 96-well plate was taken out from the cell culture incubator, the supernatant was aspirated from the well, and 30 μl of the cell lysate was added, and after 10 minutes, the ΙΟΟμΙ luciferase assay reagent was added. The ΙΟΟμΙ liquid was aspirated from each well by a pipette, added to a corresponding 96-well white plate, and the luminescence value was read on a microplate luminometer. The virus titer was calculated using the Reed-Muench method.

(3) Detection of antiviral activity of the polypeptide: The polypeptide was diluted in a ratio of 3 times (3 times) into a 96-well plate in a final volume of 50 μl, wherein 50 μl of DMEM medium was used instead of the polypeptide as a negative control. Add TZM-bl target cells (containing 15 g/ml DEAE-dextran) with a concentration of lxlO ⁵ /ml, and add 50 μl of HIV pseudovirus obtained above (100 TCID _5Q per well) at 37 °C, 5 After 48 hours of incubation under % C0 ₂ conditions, the relative fluorescence units (RLU) of each well were determined using a luciferase assay reagent (Promega). The % inhibition rate and the IC _5Q value were calculated in the same manner as described above.

 2.1.3 Peptide HP25-N2 inhibits the HIV strain NL4-3

Inhibition of HIV-1 NL4-3 replication by peptides (Chong H, Yao X, Zhang C, Cai L, Cui S, Wang Y, He Y. Biophysical Property and Broad Anti-HIV Activity of Albuvirtide. a 3- Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor. PLoS One. 2012, 7(3): e32599). The molecular cloning plasmid pNL4-3 encoding the HIV-1 strain NL4-3 was provided by the NIH AIDS Reagents and References project, catalog number 114). The pNL4-3 plasmid was prepared using QIAGEN's plasmid extraction kit, and the pNL4-3 plasmid was transfected into 293T cells using Invitrogen's LipofectamineTM 2000 transfection reagent and incubated for 6 hours at 37 ° C in a 5% C0 ₂ cell incubator. After changing the solution, the culture was continued for 48 hours. Gently collect the supernatant from the cell culture flask or cell culture plate with a pipette, filter the supernatant with a 0.45 μηι filter, add 20% FBS, and then dispense in a polypropylene tube, store at -80 ° C for use or Direct virus titration is performed in the same manner as the HIV pseudovirus described above. To determine the antiviral activity of the anti-HIV polypeptide, the polypeptide was plated in a 96-well plate at a ratio of (3 fold) dilution to a final volume of 50 μl, wherein 50 μl of DMEM medium was used instead of the polypeptide as a negative control. ΙΟΟμΙ TZM-bl cells (10 ⁵ cells/ml, containing 15 g/ml DEAE-dextran) were added, and 50 μl of the obtained virus was added, and each well was equivalent to 100 TCID _5Q . After 48 hours of culture, the relative fluorescence units (RLU) of each well were determined using a luciferase assay reagent (Promega). The % inhibition rate and the IC _5Q value were calculated in the same manner as described above.

 2.2 Experimental results and analysis

 2.2.1 Identification of highly active anti-HIV short peptides based on SC22EK

SC29EK is a C34-based polypeptide containing 29 amino acids with anti-HIV activity comparable to that of the C34 polypeptide. However, after removal of 7 amino acid residues at its C-terminus, the antiviral activity of the truncated polypeptide SC22EK was significantly reduced (Table 3, IC _{5Q in} Table 3 is the mean ± standard deviation of three replicate experiments). Compared to SC29EK, SC22EK has a severely reduced ability to inhibit HIV-mediated cell fusion, pseudoviral invasion, and wild virus replication. Based on important findings on the structure of the "MT hook", it is hypothesized that the amino acid residues (Met626 and Thr627) forming the MT hook upstream of the pocket binding region can increase the antiviral activity of the polypeptide, especially the short polypeptide. To this end, the design adds two amino acid residues, Met626 and Thr627, to SC22EK. At the N-terminus, the polypeptide MT-SC22EK in Table 3 was synthesized and tested for its viral inhibitory activity. The exciting finding is that the addition of two amino acids can significantly increase the antiviral activity of the peptide by more than 30-fold. As shown in Table 3, the IC _5Q values of MT-SC22EK inhibiting HIV-mediated cell fusion, pseudovirus invasion, and wild virus replication were 2.6 nM, 1.7 nM, and 1.2 nM, respectively, comparable to the activity of SC29EK. The activity of the SC22EK was significantly improved in the three experimental systems, which were 39.5 times, 33.9 times, and 52.1 times, respectively. This result indicates that Met626 and Thr627 amino acids in close proximity to the pocket binding region play an extremely important role in improving the anti-HIV ability of short polypeptides. This is the first time that such a short CHR polypeptide has been found to have very low nanomolar levels of anti-HIV activity. In order to find anti-HIV polypeptides with shorter sequences, the C-terminal ends were truncated with MT-SC22EK as a template, and the peptides MT-SC21EK, MT-SC20EK, MT-SC19EK, MT-SC18EK and MT-SC17EK were synthesized. 3. Their anti-HIV capabilities were quantitatively evaluated using three experimental methods. It was found that further removal of 1, 2 and 3 amino acid residues at the C-terminus had a certain effect on the activity of the polypeptide, but the corresponding three polypeptides (MT-SC21EK, MT-SC20EK and MT-SC19EK) remained unchanged. High antiviral activity. Even MT-SC19EK with 21 amino acids was significantly more active than SC22EK (containing 22 amino acids). Therefore, through this study, four highly active anti-HIV short peptides with a length of 21-24 amino acid residues containing the "MT hook" structure were found, namely MT-SC22EK, MT-SC21EK, MT-SC20EK and MT- SC19EK.

 2.2.2 Identification of highly active anti-HIV short peptides based on Ρ32 sequence truncation

The Ρ32 containing 32 amino acid residues is an anti-HIV polypeptide recently designed by the inventors of the present invention (Chinese Patent Application No.: CN201110112709. 7), and has high activity inhibitory activity against both HIV clinical isolates and Τ20 resistant viruses. The polypeptide Ν end sequence contains a Met626 Thr627 residue. In order to find a short-lived and highly active anti-HIV polypeptide, this experiment uses P32 as a template to truncate and synthesize a new set of peptides with a length shorter than or equal to 30 amino acid residues (Table 4). The anti-HIV activity of the polypeptide was examined using the above cell fusion inhibition assay and a pseudovirus invasion assay. The experimental results are shown in Table 4. First, the P32 is gradually truncated from the C-end, minus 2 (HP30), 4 (HP28), 5 (HP27), 6 (HP26), 7 (HP25), and 8 (HP24). Amino acid residues, these truncated polypeptides were found to have comparable anti-HIV activity to P32 polypeptides, including the ability to inhibit HIV-mediated cell fusion and invasion. For example, the IC _{5Q of the} polypeptide HP25 (25 amino acids in length) and HP24 (24 amino acids in length) inhibiting HIV cell fusion are Ι.ΟηΜ and l.lnM, respectively, and the IC _{5Q of} suppressing HIV pseudovirus is 0.78 nM and Ι, respectively. .ΟηΜ, and polypeptide P32 of 32 amino acid residues in length inhibited HIV cell fusion and pseudovirus 1 1. 0 n and 0.92 nM, respectively. At the same time, the anti-HIV activities of the polypeptides C34, CP32M, Sifuvirtide, T1249 and T20 as controls were significantly lower than those of P32 and its above-described truncated polypeptide. However, further truncation of the C-terminus (HP23, HP22 and HP19) would result in a significant reduction or complete loss of the polypeptide's inhibitory activity against the virus (Table 4).

Table 4. Inhibitory activity of polypeptides against HIV-1 mediated cell fusion and pseudoviral invasion (IC _5Q , nM)

Peptide amino acid sequence polypeptide length cell fusion pseudovirus : truncated peptide

 ΪΗΡ30 NEMTWEEWEKKIEEYTKKIEEILKKSEEQ 30 1.13 0.91

ΪΗΡ28 NEMTWEEWEKKIEEYTKKIEEILKKSE 28 1.05 0.9

ΪΗΡ27 NEMTWEEWEKKIEEYTKKIEEILKKS 27 1.2 0.96

ΪΗΡ26 NEMTWEEWEKKIEEYTKKIEEILKK 26 1.19 0.93

ΪΗΡ25 NEMTWEEWEKKIEEYTKKIEEILK 25 1 0.78

ΪΗΡ24 NEMTWEEWEKKIEEYTKKIEEIL 24 1.1 1

ΪΗΡ23 NEMTWEEWEKKIEEYTKKIEEI 23 552.42

ΪΗΡ22 NEMTWEEWEKKIEEYTKKIEE 22 >1000 >1000 HP19 NEMTWEEWEKKIEEYTKK 19 >1000 >1000

ΪΗΡ24-Ν1 NEMTWEEWEKKIEEYTKKIEEIL 23 1.97 1.69 HP24-N2 EMTWEEWEKKI ΕΕΥΤΚΚΙ EEIL 22 1.42 1.46 HP24-N3 MTWEEWEKKIEEYTKKIEEIL 21 1.93 2.06 HP24-N4 TWEEWEKKIEEYTKKIEEIL 20 40.77 52.81 HP24-N5 WEEWEKKIEEYTKKIEEIL 19 68.3 106.62 i

ΪΗΡ27-Ν3 MTWEEWEKKIEEYTKKIEEILKKS 24 1.54 1.18 HP26-N3 MTWEEWEKKIEEYTKKIEEILKK 23 1.8 1.36

ΪΗΡ25-Ν3 MTWEEWEKKIEEYTKKIEEILK 22 1.23 0.86

ΪΗΡ25-Ν2 EMTWEEWEKKI EEYTKKI EE ILK 23 0.74 0.58

:ELT23 \ ELTWEEWEKKI EEYTKKI EE ILK 23 0.9 0.61

Control peptide

 ΪΡ32 EMTWEEWEKKI EEYTKKI EE I LKKSEEQQK 32 1.21 0.92

:C34 \ ME DREINNYTSLIHSLIEESQNQQEKNEQELL 34 1.47 2.62

:CP32M \ VE NEMT MEWEREIENYTKLIYKILEESQEQ 32 4.35 2.83 Sifuvirtide \ SWETWEREIENYTKQIYKILEESQEQQDRNEKDLLE 36 2.88 2.78

:T1249 \ WQEWEQKITALLEQAQIQQEKNEYELQKLDK ASLWEWF \ 39 3.73 1.98

ΪΤ2635 \ TTWEAWDRAIAEYAARI EALIRAAQEQQEKNEAALREL \ 39 1.02 1.03

:Τ20 \ YTSLIHSLIEESQNQQEKNEQELLELDK ASL WF 36 10.66 70.45 In order to find a shorter anti-HIV polypeptide, this experiment uses HP24 as a template to cut the N-terminus one by one, and remove one (HP24-N1) and two (HP24-N2) respectively. , 3 (HP24-N3), 4 (HP24-N4) and 5 (HP24-N5) amino acid residues. It was found that the activity of HP24-N2 was slightly affected, and the activities of HP24-N1 and HP24-N3 decreased nearly 1-fold, but the activities of HP24-N4 and HP24-N5 were significantly decreased. Once again, the N-terminal amino acid residues Met626 and Thr627 play a key role in the antiviral activity of these short polypeptides.

Furthermore, this experiment truncated HP27, HP26 and HP25 as templates, and found that the three amino acid residues (WNE) at the N-terminus did have a limited effect on the antiviral activity of the polypeptide, but the surprising discovery was that HP25-N2 The activity is significantly higher than other polypeptides. Compared with the template HP25, HP25-N2 was further improved in anti-HIV cell fusion and pseudovirus invasion activity, with IC _5Q of 0.74 nM and 0.58 nM, respectively, which was significantly lower than other truncated polypeptides and all control polypeptides. In order to avoid the problem that the side chain of methionine is easily oxidized, a similarly long leucine (L) was substituted for methionine (M), and a polypeptide of the same length of 23 amino acids ELT23 was synthesized (see Table 4 for the sequence). The results showed that ELT23 has similar antiviral activity to HP25-N2, and its IC50 for inhibiting viral cell fusion and invasion is 0.9 nM and 0.61 nM, respectively.

To further confirm the antiviral activity of HP25-N2, its inhibition of HIV strain NL4-3 replication was also tested, and the polypeptides MT-SC22EK, MT-SC29EK, Sifuvirtide, C34 and T20 were used as controls. The results of the experiment are shown in Figure 1. The inhibitory activity of T20 against HIV-1 _NM _ ₃ strain is relatively low, and its IC _5Q is 54. InM; MT-SC22EK, MT-SC29EK, Sifuvirtide and C34 activity comparison Closely, IC ₅ Q was 1.2 nM, 1.0 nM, 1.8 nM, and 1.3 nM, respectively; in contrast, HP25-N2 had significantly higher anti-HIV activity with an IC ₅₀ of 0.3 nM o

Therefore, through intensive research, the inventors identified for the first time a group of highly active anti-HIV polypeptides ranging in length from 21 to 30 amino acid residues, including HP30, HP28, HP27, HP26, HP25, HP24, HP24-N1, HP24- N2, HP24-N3, HP25-N2, HP25-N3, HP27-N3 HP26-N3, ELT23 (sequence and activity are shown in Table 4 and Figure 1, IC _5Q in Table 4 is the average of three replicate experiments, The data in 1 is the average of three replicate experiments). In contrast, HP25-N2, which has a length of 23 amino acids, and its derivative ELT23 have better antiviral activity.

 Example 3. HP25-N2 broad spectrum anti-HIV effect

Because HIV-1 is susceptible to variability, it can be divided into a variety of subtypes, including AD, FH, J, and K subtypes. Among them, subtypes A, B and C are the main viruses causing the world AIDS epidemic. In China, the prevalence of B/C and A/E recombinant viruses is dominant. To further evaluate the activity of anti-HIV short peptides, a group of 16 HIV pseudoviruses were prepared, including international representative strains and HIV strains currently prevalent in China, including 2 subtypes A, 5 subtype B, and C. There were 3 strains, 3 A/E recombinant strains, and 3 B/C recombinant strains. Preparation of pseudoviruses As described above, HIV envelope protein granules expressing various subtypes are preserved in the laboratory of Prof. He Yuxian, Institute of Pathogenic Biology, Chinese Academy of Medical Sciences, see literature (Yao X, Chong H, Zhang C, Waltersperger S, Wang M, Cui S, He Y. Broad antiviral activity and crystal structure of HIV-1 fusion inhibitor Sifuvirtide. J Biol Chem. 2012, 287: 6788-6796). Using these HIV pseudoviruses, the antiviral of HP25-N2 (sequence 14 in the sequence listing) was tested and the C34 polypeptide was used as a control. The experimental results are shown in Table 5 (IC _{5Q in} Table 5 is the mean standard deviation of three replicate experiments), and HP25-N2 is effective in inhibiting infection of various HIV subtypes. For most viruses, the IC _5Q value is lower than C34. For example, HP25-N2 inhibits the subtype B strain SF162 with an IC _5Q value of 3.6 nM, and C34 has an IC _5Q value of 9.5 nM for the strain. Table 5. Inhibition of different subtypes of Hl g by the polypeptide HP25-N2

IC ₅₀ (nM)

 HIV pseudovirus subtype HP25-N2 C34

92UG037.8 A 1.3 ±0.2 3.0 ±0.5

92RW020 A 1.5 ±0.2 3.1 ±0.8

SF162 B 3.6 ±0.7 9.5 ±1.4

AC10.0.29 B 1.0 ±0.1 1.5 ±0.4

TRO.11 B 3.1 ±0.5 4.4 ±1.2 pREJ04541 B 0.9 ±0.2 0.7 ±0.1

SC422661.8 B 0.8 ±0.1 0.5 ±0.0

CAP45.2.00 C 2.8 ±0.3 13.2 ±1.1

ZM214M.PL15 C 1.7 ±0.5 3.4 ±0.6

ZM109F.PB C 0.8 ±0.1 1.8 ±0.3

AE01 A/E 1.0 ±0.1 2.7 ±0.1

AE03 A/E 1.6 ±0.3 3.0 ±0.7

GX11.13 A/E 3.9 ±0.8 8.2 ±1.1

CH64.20 B/C 0.6 ±0.1 1.1 ±0.1

CH070.1 B/C 3.0 ±0.3 5.1 ±0.4

Ch120.6 B/C 3.5 ±0.6 3.5 ±0.8 Example 4. Inhibition of HP25-N2 against T20-resistant HIV-1 strain

T20 is currently the only HIV membrane fusion inhibitor approved for clinical treatment. However, its activity is not only significantly lower than that of a new generation of peptides, but also easily induces drug-resistant mutations, leading to the failure of clinical antiviral therapy. Studies have shown that amino acid variation in the NHR region of drug target sequences is the main cause of T20 resistance. The development of new inhibitors that inhibit T20-resistant viruses is currently a hot topic internationally. This example observes the effect of HP25-N2 (sequence 14 in the Sequence Listing) against T20 and C34 resistant HIV strains. Pseudoviruses carrying NHR mutations (Table 6, IC _{5Q in} Table 6 is the mean ± standard deviation of three replicate experiments) using HIV mutant strains reported in the literature (Chong H, Yao X, Zhang C, Cai L, Cui S , Wang Y, He Y. Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor. PLoS One.2012, 7(3): e32599.). The inhibitory effect of the polypeptide was evaluated by the above-described HIV pseudovirus technology system. The preparation and application method of the pseudovirus is the same as that of the second embodiment. From the experimental results in Table 6, it is known that the anti-HIV polypeptide HP25-N2 has extremely high activity against these HIV strains carrying the gp41 mutation, indicating that these mutations have little effect on the antiviral activity of HP25-N2. However, the activity of T20 as a control was significantly decreased, showing extremely high drug resistance. For example, the resistance multiplicity of the V38A mutation was 34.2, and the resistance ratio of the I37T/N43K and V38A/N42T double mutation was above 44. Moreover, these resistance mutations also led to significant cross-resistance of C34. The results of this experiment indicate that HP25-N2 peptide can be used for the treatment of T20 clinically resistant HIV strains.

Table 6. Inhibition of HP25-N2 against T20 and C34 resistant HIV strains T20 C34 HP25-N2

:HIV-1 _NL4 . ₃ ; IC ₅₀ (nM) : Fold change; IC ₅₀ (nM) Fold change: IC ₅₀ (nM) ; Fold change; iWT : 51.1 ±12.4 ; 1 1.2 ±0.2 1 0.7 ±0.1 1

M37T : 540.2 ±135.3; 10.6 ; 25.0 ±2.8 20.8 0.7 ±0.0 1

;V38A : 1746.3 ±80.4; 34.2 ; 28.7 ±3.4 23.9 0.7 ±0.0 1

;V38M : 370.2 ±61.2 ; 7.2 ; 18.5 ±2.8 15.4 0.9 ±0.1 1.2

;Q40H Μ242·0± 120.4; 24.3 : 67.8 ±7.2 56.5 0.7 ±0.1 1

N43K ; 316.9 ±51.3 : 6.2 : 17.3 ±1.4 14.4 0.7 ±0.1 1

; D36S/V38M ; 187.2 ±13.9 : 3.7 : 7.1 ±1.4 6 1.2 ±0.1 1.7

M37T7N43K >2250 >44 : 426.4 ±18.2 355.3 0.9 ±0.0 1.3

V38A/N42T >2250 >44 : 257.7 ±19.7 214.8 0.3 ±0.0 0.4 Example 5. Circular dichroism (CD) technique studies the interaction of peptides with NHR targets

 5.1 Experimental materials and methods

 Experimental Methods References (Chong H, Yao X, Zhang C, Cai L, Cui S, Wang Y, He Y. Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor. PLoS One .2012, 7(3): e32599; Chong H, Yao X, Qiu Z, Qin B, Han R, Waltersperger S, Wang M, Cui S, He Y. Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652. J Biol Chem. 2012, 287(24): 20281-20289). The circular dichrometer is Nissan Jasco-815. The N36 polypeptide derived from NHR was used as a target sequence, and its sequence was: Ac-SGIVQQQ NLLRAIEAQQHLLQLTVWGIKQLQARIL-NH2. The CHR-derived antiviral polypeptide and N36 were separately dissolved in phosphate buffered saline (PBS), and the concentration was determined by ultraviolet absorption at 280 nm, and then 20 μL of the polypeptide PBS solution (pH 7.2) was prepared. The antiviral polypeptide was mixed with N36 in a volume ratio of 1:1 to obtain a mixed sample (the final concentration of each peptide was 10 μM), and the sample was allowed to stand at 37 ° C for 30 minutes to fully react. The prepared sample was measured on a circular dichroic instrument with a scanning wavelength range of 195-260 nm, a wavelength interval of 1 nm, and an average of 3 scans. The measurement was carried out at room temperature. The blank is scanned with a PBS buffer solution, and then the sample signal is scanned, and the blank signal is subtracted from the sample signal to obtain a CD signal. The interaction between the polypeptides and the helix content were determined based on the CD signal. The stability of the six-helix structure formed by the antiviral polypeptide described herein and N36 was determined by CD temperature scanning. The specific method is as follows: The above polypeptide for measuring CD signal is transferred into a sample cell (can also be reconstituted), and the CD instrument program is set to a temperature scan, the detection wavelength is 220 nm, the scanning range is 20-98 ° C, and the program temperature is scanned. The curve of the CD signal as a function of temperature is obtained. Calculate the m value from the curve.

 5.2 Experimental results and analysis

The antiviral polypeptide derived from the CHR region of HIV gp41 mainly blocks the formation of the 6-HB structure by interacting with the target sequence of the NHR region, thereby inhibiting the invasion of HIV into target cells. The results of the CD test showed that all anti-HIV polypeptides formed a typical helical structure with the NHR polypeptide N36, showing a typical absorption peak at -208 and -222 (as shown in Figures 2, 3 and 7). Among them, the greater the negative value of [θ] 222, the higher the α-helix content of the CHR and NHR polypeptide complexes. In comparison, based on MT-SC17EK and The 6-HB of MT-SC18EK has a low helix content, and its [θ]222 values are -21.3 and -17.5, respectively; the short peptides 22 and HP19 based on the Ρ32 design have a relatively low helix content, [θ The value of 222 is -17.1 and -15.4, respectively. The four peptides were not observed to inhibit HIV activity in the above antiviral experiments (greater than 100 OnM). The results of the CD experiment found that the Tm value of MT-SC22EK and N36 (78.2 °C) was higher than the m value of SC22EK and N36 (64.1). A significant increase, also well above the m value of SC29EK and N36 (67.1), indicates that the "MT construct" amino acid residues, Met626 and Thr627, do significantly increase the stability of the interaction of the antiviral polypeptide with the target sequence. It was found that the anti-viral activity of the polypeptide HP25-N2 has a relatively high helix content ([θ] ₂₂₂ = -28.8 ) and 6-HB stability (m = 89.2 °C). In addition, the m of the four anti-HIV active polypeptides (MT-SC17EK MT-SC18EK HP22 and HP19) were significantly lower than the other polypeptides, 51.1 °C, 56.1 °C, 53.1 °C and 49.1 °C, respectively. These results indicate that the viral inhibitory activity of the anti-HIV polypeptide is related to the helicity and thermostability of the heterologous 6-HB.

 Table 7. Circular dichroism analysis of the interaction of antiviral polypeptides with N36

注： 表中多肽的氨基酸序列如表 3和表 4。

Note: The amino acid sequences of the peptides in the table are shown in Tables 3 and 4.

Industrial application

 The inventors of the present invention found that the addition of two amino acid residues (Met626 and Thr627) forming the "M-T hook" at the N-terminus of the short peptide SC22EK can significantly improve its antiviral activity and 6-HB stability. Furthermore, by truncating the P32 polypeptide sequence, it was found that a group of short polypeptides of 21-30 amino acid residues in length have a strong inhibitory effect on HIV. These polypeptides, although significantly shorter than other polypeptides reported, can stably bind to their NHR target sequences. The polypeptides provided by the present invention have significant advantages:

 (1) Containing only 21-30 amino acid residues, significantly shorter than other reported polypeptides, such as T20 and Sifuvirtide (both 36 amino acids), T1249 and T2635 (all 39 amino acids), C34 and SC34EK (both 34 amino acids) and so on. Therefore, the polypeptide of the present invention has the advantage of being easy to synthesize and low in cost.

 (2) The polypeptide of the present invention is significantly shorter than other polypeptides but has extremely strong antiviral activity, for example, the activity of inhibiting HIV is significantly higher than that of T20, T1249 and sifuvirtide.

 (3) The polypeptide of the present invention retains extremely strong activity against the Τ20 resistant strain. For example, the peptide ΗΡ25-Ν2 can completely overcome many clinically common Τ20 resistant HIV mutant strains.

 (4) The polypeptide of the present invention has a "salt bridge structure" (ΕΚ site) which can enhance the helicity of the polypeptide by containing an amino acid residue at the end of the sputum which can stabilize the interaction of the polypeptide with the target and forming a "hook structure". Therefore, the target sequence can be more firmly bound, thereby having superior stability.

 (5) The polypeptide of the present invention is easily soluble in water due to the large introduction of hydrophilic polar amino acids (e.g., ruthenium and osmium) in the polypeptide sequence. The improvement of water solubility facilitates the development of drug efficacy and the development of pharmaceutical dosage forms.

 The invention will lay a solid foundation for the development of a new generation of AIDS treatment and prevention drugs.

Claims

Rights request A polypeptide, a pharmaceutically acceptable salt thereof, or a derivative thereof, characterized in that: the polypeptide is as follows a) or b) or c):  a) the polypeptide HP25-N2 shown in SEQ ID NO: 14 in the sequence listing;  b) a derivative polypeptide having anti-HIV activity obtained by adding more than one amino acid residue at the amino terminus and/or the carboxy terminus of the polypeptide of SEQ ID NO: 19, the derivatized polypeptide consisting of 21-30 amino acid residues;  c) a derivative polypeptide consisting of 21-30 amino acid residues obtained by adding or substituting one or more amino acids at any position including the amino terminus and the carboxy terminus of the polypeptide of SEQ ID NO: 19 in the sequence listing, the derived polypeptide having Anti-HIV activity.  The polypeptide, the pharmaceutically acceptable salt thereof or the derivative thereof according to claim 1, wherein the derivative polypeptide represented by b) is any one of the following bl) - bl7):  Bl) the amino acid sequence is the polypeptide of sequence 18 in the sequence listing ELT23;  B2) The amino acid sequence is the polypeptide of sequence 9 in the sequence listing HP25 ;  B3) the amino acid sequence is the polypeptide of sequence 6 in the sequence listing, HP28;  B4) the amino acid sequence is the polypeptide of sequence 10 in the sequence listing, HP24;  B5) the amino acid sequence is the polypeptide of sequence 5 in the sequence listing HP30;  B6) the amino acid sequence is the polypeptide of sequence 8 in the sequence listing, HP26;  B7) the amino acid sequence is the polypeptide of sequence 7 in the sequence listing HP27;  B8) the amino acid sequence is the polypeptide of sequence 15 in the sequence listing HP25-N3;  B9) the amino acid sequence is the polypeptide of sequence 12 in the sequence listing, HP24-N2;  blO) the amino acid sequence is the polypeptide of sequence 17 in the sequence listing HP27-N3;  Bl l ) amino acid sequence is the polypeptide of sequence 16 in the sequence listing HP26-N3;  Bl2) the amino acid sequence is the polypeptide of sequence 13 in the sequence listing, HP24-N3;  Bl3 ) the amino acid sequence is the polypeptide of sequence 11 in the sequence listing HP24-N1 ;  Bl4) the amino acid sequence is the polypeptide of sequence 1 in the sequence listing MT-SC22EK;  Bl5) the amino acid sequence is the polypeptide of sequence 2 in the sequence listing MT-SC21EK;  Bl6) the amino acid sequence is the polypeptide of sequence 3 in the sequence listing MT-SC20EK;  Bl7) The amino acid sequence is the polypeptide MT-SC19EK of sequence 4 in the Sequence Listing.  The polypeptide, the pharmaceutically acceptable salt thereof or the derivative thereof according to claim 1 or 2, wherein the polypeptide derivative according to claim 1 or 2 is at least one of the following 1) to 5) :  1) a linker obtained by linking the amino terminus of the polypeptide of claim 1 or 2 to the amino terminal protecting group and/or the carboxy terminal of the polypeptide of claim 1 or 2 to the carboxy terminal protecting group;  2) a carboxy terminal of the polypeptide according to claim 1 or 2 which is linked to an oligopeptide or a lipophilic group or a cholesterol-derived linker; 3) The amino terminus of the polypeptide of claim 1 or 2 is linked to an oligopeptide or a lipophilic group or cholesterol Connector  4) A linker obtained by linking an oligopeptide or a lipophilic group or cholesterol to both the amino terminus and the carboxy terminus of the polypeptide of claim 1 or 2;  5) A modification of the polypeptide according to claim 1 or 2 which is modified with a protein, polyethylene glycol or maleimide.  The polypeptide, the pharmaceutically acceptable salt thereof or the derivative thereof according to claim 3, wherein the polypeptide derivative according to claim 1 or 2 is: the amino terminal of the polypeptide of claim 1 or A linker obtained by linking an amide group to the carboxy group of the polypeptide of claim 1 or 2.  A polymer formed from the polypeptide of any one of claims 1 to 4 and/or a pharmaceutically acceptable salt thereof and/or a derivative thereof.  6. A composition comprising C1) and C2): C1) a polypeptide according to any one of claims 1 to 4, a derivative thereof, or a pharmaceutically acceptable salt thereof, and/or according to claim 5 a polymer; C2) a pharmaceutically acceptable carrier or adjuvant;  The composition has at least one of the following D1) - D5):  D1) anti-HIV;  D2) treatment and / or prevention and / or adjuvant treatment of diseases caused by HIV infection (such as AIDS);  D3) inhibiting HIV for cell fusion;  D4) inhibit HIV from invading cells;  D5) inhibit HIV replication.  The polypeptide according to any one of claims 1 to 4, a derivative thereof, or a pharmaceutically acceptable salt thereof, or the multimer according to claim 5, or the composition according to claim 6 is prepared as follows Application in one product: D1) anti-HIV products, such as drugs or vaccines;  D2) products that treat and/or prevent and/or adjuvant treatment of diseases caused by HIV infection, such as drugs or vaccines; D3) products that inhibit HIV cell fusion;  D4) Products that inhibit HIV from invading cells;  D5) Products that inhibit HIV replication.  8. A nucleic acid molecule encoding the polypeptide of any one of claims 1 to 4.  9. The method of preparation of the product, including el) or e2):  El) a step of chemically synthesizing the polypeptide of any of claims 1-4;  E2) a step of biologically expressing the polypeptide of any of claims 1-4; The product is fl), £2) or β) _:  The polypeptide according to any one of claims 1 to 4, a pharmaceutically acceptable salt thereof, or a derivative thereof;  £2) the multimer of claim 5;  O) The composition of claim 6.  10. Biomaterials of gl) or g2): Gl) an expression cassette, recombinant vector, recombinant cell or recombinant virus comprising the nucleic acid molecule of claim 8; g2) a recombinant expression vector or recombinant cell expressing the polypeptide of any of claims 1-4. 11. A method of treating and/or preventing and/or adjuvant treatment of an enveloped viral infection, comprising the step of administering to the subject an effective amount of fl) and / or /f2) and / or beta):  Omega) a polypeptide according to any one of claims 1 to 4, a pharmaceutically acceptable salt thereof, or a derivative thereof;  Ω) the multimer of claim 5;  β) The composition of claim 6.