Lipopeptide HIV membrane fusion inhibitor and pharmaceutical application

Abstract

The invention discloses a lipopeptide HIV (Human Immunodeficiency Virus) membrane fusion inhibitor and a pharmaceutical application. The lipopeptide disclosed by the invention is shown as a formula I: X < 1 >-EMTWEEK KVEELEKIEEELK-X < 2 >-X < 3 >-X < 4 >-X < 5 >, or a formula II: X < 1 >-ELTWEEWEKKVEELEKIEEELK-X < 6 >-X < 7 >-X < 4 >-X < 5 >; wherein X1 is an amino-terminal protecting group; x2 is a polypeptide sequence (EAAAK) n, A [(EAAAK) n] A or (EP) n, and n represents a repetition number; x3 is Lys, Cys, Dap, Orn, Dab, Dah or does not exist; x4 is a lipophilic compound group for modifying the C terminal; x5 is a carboxyl end protecting group; x6 is a polypeptide sequence KAEEQQKKNE, and X7 is Lys, Cy s, Dap, Orn, Dab or Dah. The lipopeptide provided by the invention has higher antiviral activity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a lipopeptide-based HIV membrane fusion inhibitor and a pharmaceutical use thereof. Background Art

[0002] Acquired Immune Deficiency Syndrome (AIDS) is a chronic disease caused by human immunodeficiency virus (HIV). HIV mainly infects CD4+T cells and dendritic cells expressing CD4 molecules, monocytes and macrophages, etc., attacks the human immune system, causes opportunistic infections and tumors, and seriously threatens the life and health of patients. However, there is no effective vaccine or cure so far, and AIDS is still a major global public health issue.

[0003] Currently approved anti-HIV drugs mainly target four stages of the viral life cycle, including viral invasion, reverse transcription, integration, and maturation. The process of HIV invading target cells is mediated by the envelope glycoprotein (Env) on its surface, which is composed of the surface subunit gp120 and the transmembrane subunit gp41 connected by non-covalent bonds. When the virus infects the target cell, the surface subunit gp120 of the envelope glycoprotein first binds to the CD4 receptor on the target cell membrane, and the conformation of gp120 changes, exposing its binding site with the auxiliary receptor; subsequently, gp120 binds to the auxiliary receptor (CCR5 or CXCR4) on the surface of the target cell, causing it to split from the transmembrane subunit gp41; finally, the conformation of gp41 changes, initiating the fusion of the viral envelope with the target cell membrane, completing the process of the virus entering the host cell. In terms of mechanism of action, HIV fusion inhibitors can competitively bind to the N- or C-terminal heptad repeat region (NHR or CHR) of gp41, thereby blocking the formation of the six-helix bundle (6-HB) structure and inhibiting the virus from entering the target cell. T20 is the first and currently the only HIV fusion inhibitor drug approved by the US FDA for marketing. It is derived from amino acids 638 to 673 of gp41CHR and consists of 36 amino acid residues. However, due to its relatively low antiviral activity, it requires large-dose injections (twice a day, 90 mg each time), and has defects such as a short half-life, easy to develop drug resistance, and high treatment costs, its clinical application is significantly limited. Therefore, it is crucial to develop new HIV fusion inhibitors with improved drug properties.

[0004] The inventors' previous studies have shown that the MT hook structure can significantly enhance the binding and antiviral activity of CHR-derived peptide inhibitors, and based on this, they designed a short peptide membrane fusion inhibitor 2P23 containing the MT hook structure and the PBD region sequence. 2P23 has potent and broad-spectrum antiviral activity against HIV-1, HIV-2 and SIV; in order to further improve the stability and antiviral activity of 2P23, palmitic acid (C16)-modified lipopeptide LP-19 and stearic acid (C18)-modified lipopeptide LP-25 were subsequently prepared. Both lipopeptides showed efficient anti-HIV activity in vitro and in vivo. Many studies have shown that cholesterol modification is also an important strategy for the development of lipopeptide-based viral fusion inhibitors, such as HIV fusion inhibitory lipopeptides C34-Chol, LP-98, etc. A series of SARS-CoV-2 fusion-inhibiting lipopeptides were rapidly developed through cholesterol modification, including SARS-CoV-2HR2-derived lipopeptides IPB02, IPB24, IPB29, SARSHRC-PEG4-chol, P40-LP and P315V3, and HCoV-OC43 HR2-derived lipopeptide EK1C4. In the lipopeptide-based design strategy, it is usually necessary to add a linker between the lipid molecule and the polypeptide sequence to act as a connecting arm. The flexible linker small molecule polyethylene glycol (PEG)n and the combination of glycine (G) and serine (S), such as (GGGGS)n or (GSGSG)n, have been widely used. Summary of the invention

[0005] In order to improve the antiviral activity of HIV membrane fusion inhibitors, the present invention aims to provide lipopeptide HIV membrane fusion inhibitors and pharmaceutical uses.

[0006] The specific technical solutions of the present invention are as follows:

[0007] The present invention provides a lipopeptide or a pharmaceutically acceptable salt thereof, characterized in that the lipopeptide is represented by the following formula I or formula II:

[0008] Formula I: X1-EMTWEEWEKKVEELEKKIEELLK-X2-X3-X4-X5;

[0009] Formula II: X1-ELTWEEWEKKVEELEKKIEELLK-X6-X7-X4-X5;

[0010] Wherein, X1 is an amino terminal protecting group, X4 is a lipophilic compound group used to modify the C-terminus, and X5 is a carboxyl terminal protecting group;

[0011] The X2 and X3 are any one of the following 1) and 2):

[0012] 1) X2 is a polypeptide sequence (EAAAK)n, which acts as a rigid linker and serves as a connecting arm, wherein n represents the number of repetitions of EAAAK, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine (Lys), cysteine ​​(Cys), 2,3-diaminopropionic acid (Dap), ornithine (Orn), 2,4-diaminobutyric acid (Dab), 2,7-diaminoheptanoic acid (Dah) or does not exist;

[0013] 2) X2 is a polypeptide sequence A[(EAAAK)n]A or (EP)n, which acts as a rigid linker and serves as a connecting arm, wherein n represents the number of repetitions of EAAAK or EP, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine (Lys), cysteine ​​(Cys), 2,3-diaminopropionic acid (Dap), ornithine (Orn), 2,4-diaminobutyric acid (Dab) or 2,7-diaminoheptanoic acid (Dah);

[0014] The X6 is a polypeptide sequence KAEEQQKKNE, and the X7 is lysine (Lys), cysteine ​​(Cys), 2,3-diaminopropionic acid (Dap), ornithine (Orn), 2,4-diaminobutyric acid (Dab) or 2,7-diaminoheptanoic acid (Dah).

[0015] Furthermore, the amino terminal protecting group is any one of acetyl (Ac), amino (NH2), maleyl, succinyl, tert-butyloxycarbonyl or benzyloxy or other hydrophobic groups or macromolecular carrier groups.

[0016] Furthermore, the carboxyl terminal protecting group is any one of amino (NH2), carboxyl, hydroxyl, amide or tert-butyloxycarbonyl or other hydrophobic groups or macromolecular carrier groups.

[0017] Further, the lipophilic compound is any one of cholesterol succinate, 2-cholesterol acetic acid, 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, fatty acids containing 8 to 20 carbon atoms (such as octadecanoic acid), dihydrosphingosine, and vitamin E, and is preferably cholesterol succinate.

[0018] Furthermore, the amino terminal protecting group is an acetyl group; and / or the carboxyl terminal protecting group is an amino group.

[0019] Furthermore, X2 and X3 are any one of the following 1) and 2):

[0020] 1) X2 is a polypeptide sequence (EAAAK)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine or does not exist;

[0021] 2) X2 is a polypeptide sequence A[(EAAAK)n]A or (EP)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine;

[0022] The X7 is lysine.

[0023] Furthermore, the compound of formula I is LP-35, X1 is Ac, X2 is EAAAK (n=1), X3 is lysine, X4 is cholesterol succinate, and X5 is NH2.

[0024] Furthermore, the compound of formula I is LP-36, X1 is Ac, X2 is EAAAKEAAAK (n=2), X3 is lysine, X4 is cholesterol succinate, and X5 is NH2.

[0025] Furthermore, the compound of formula I is LP-37, X1 is Ac, X2 is AEAAAKEAAAKA (n=2), X3 is lysine, X4 is cholesterol succinate, and X5 is NH2.

[0026] Further, the compound of formula I is LP-38, X1 is Ac, X2 is EAAAKEAAAKEAAAK (n=3), X3 is absent, X4 is cholesterol succinate, and X5 is NH2.

[0027] Furthermore, the compound of formula I is LP-39, X1 is Ac, X2 is EPEPEPEPEPEP (n=6), X3 is lysine, X4 is cholesterol succinate, and X5 is NH2.

[0028] Furthermore, the compound of formula II is LP-32, X1 is Ac, X6 is KAEEQQKKNE, X7 is lysine, X4 is cholesterol succinate, and X5 is NH2.

[0029] The abbreviations of amino acids in Formula I and Formula II have meanings well known in the art, for example, E is glutamic acid, M is methionine, T is threonine, W is tryptophan, K is lysine, V is valine, L is leucine, I is isoleucine, P is proline, etc.

[0030] The present invention also provides a pharmaceutical composition, which comprises the lipopeptide or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0031] The present invention also provides use of the lipopeptide or a pharmaceutically acceptable salt thereof or a pharmaceutical composition in preparing a lipopeptide HIV membrane fusion inhibitor.

[0032] The present invention also provides use of the lipopeptide or a pharmaceutically acceptable salt thereof or a pharmaceutical composition in the preparation of any of the following drugs:

[0033] 1) Antiviral drugs;

[0034] 2) Drugs for preventing and / or treating diseases caused by viral infections;

[0035] 3) Drugs that inhibit virus-cell fusion;

[0036] 4) Drugs that inhibit viruses from invading cells;

[0037] 5) Drugs that inhibit viral replication;

[0038] Wherein, the virus is one or more of HIV-1, HIV-2 and SIV.

[0039] Furthermore, the disease caused by the viral infection is AIDS.

[0040] More contents involved in the present invention are described in detail below, or some of them can also be embodied in the embodiments of the present invention.

[0041] Unless otherwise indicated, the quantities of different components and reaction conditions used herein may be interpreted as "roughly" or "approximately" in any case. Accordingly, unless otherwise specified, the numerical parameters cited below and in the claims are approximate parameters, and different numerical parameters may be obtained under respective experimental conditions due to different standard errors.

[0042] Herein, when there is a disagreement or ambiguity between the chemical formula and chemical name of a compound, the chemical formula is used to accurately define the compound. The compounds described in the present invention may contain one or more chiral centers, and / or double bonds and structures such as these, and may also have stereoisomers, including double bond isomers (such as geometric isomers), optical enantiomers or diastereomers. Accordingly, any chemical structure within the scope of the description herein, whether it contains the above-mentioned similar structures in part or in its entirety, includes all possible enantiomers and diastereomers of the compound, including any single stereoisomer (such as a single geometric isomer, a single enantiomer or a single diastereomer) and any mixture of these isomers. These racemic isomers and mixtures of stereoisomers can also be further separated into their constituent enantiomers or stereoisomers by those skilled in the art using different separation techniques or methods for synthesizing chiral molecules.

[0043] The compounds of formula I and formula II include, but are not limited to, optical isomers, racemates and / or other mixtures of these compounds. In the above cases, single enantiomers or diastereomers, such as optically active isomers, can be obtained by asymmetric synthesis or racemate resolution. The resolution of the racemate can be achieved by different methods, such as conventional recrystallization with a reagent that assists resolution, or by chromatographic methods. In addition, the compounds of formula I and formula II also include cis and / or trans isomers with double bonds.

[0044] The compounds of the present invention include, but are not limited to, compounds of Formula I and Formula II and all of their pharmaceutically acceptable different forms. The pharmaceutically acceptable different forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above substances, and any mixtures of the above forms.

[0045] In practical applications, the medicine of the present invention can be directly administered to a patient, or mixed with a suitable carrier or excipient to achieve the purpose of treating and / or preventing HIV infection. The carrier material here includes, but is not limited to, water-soluble carrier materials (such as polyethylene glycol, polyvinyl pyrrolidone, organic acids, etc.), poorly soluble carrier materials (such as ethyl cellulose, cholesterol stearate, etc.), and enteric carrier materials (such as cellulose acetate phthalate and carboxymethyl ethyl cellulose, etc.). Among them, water-soluble carrier materials are preferred. These materials can be made into a variety of dosage forms, including but not limited to tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections, etc. It can be a common preparation, a sustained-release preparation, a controlled-release preparation, and various microparticle delivery systems. In order to make the unit dosage form into tablets, various carriers known in the art can be widely used. Examples of carriers include 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, glycerol, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, acacia paste, gelatin paste, sodium carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinyl pyrrolidone, etc.; disintegrants. , such as dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid esters, sodium lauryl sulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors, such as sucrose, tristearate, cocoa butter, hydrogenated oil, etc.; absorption promoters, such as quaternary ammonium salts, sodium lauryl sulfate, etc.; lubricants, such as talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc. The tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets. In order to make the unit dosage form into a pill, various carriers known in the art can be widely used. Examples of carriers include diluents and absorbents, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinyl pyrrolidone, Gelucire, kaolin, talc, etc.; binders, such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste or flour paste, etc.; disintegrants, such as agar powder, dry starch, alginate, sodium lauryl sulfate, methyl cellulose, ethyl cellulose, etc. In order to prepare a unit dosage form as a suppository, various carriers known in the art can be widely used. Examples of carriers include, for example, polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, etc.In order to prepare the unit dosage form into an injectable preparation, such as a solution, emulsion, freeze-dried powder injection and suspension, all diluents commonly used in the art can be used, for example, water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isostearyl alcohol, polyoxyethylene sorbitol fatty acid esters, etc. In addition, in order to prepare an isotonic injection, an appropriate amount of sodium chloride, glucose or glycerol can be added to the injectable preparation. In addition, conventional cosolvents, buffers, pH regulators, etc. can also be added. In addition, if necessary, colorants, preservatives, spices, flavoring agents, sweeteners or other materials can also be added to the pharmaceutical preparation. The above dosage form can be administered by injection, including subcutaneous injection, intravenous injection, intramuscular injection and intracavitary injection, etc.; cavity administration, such as rectal and vaginal. The above-mentioned route of administration is preferably injection.

[0046] The dosage of the drug of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight and individual response of the patient or animal, the specific active ingredient used, the route of administration and the number of administrations, etc. The above dosage can be administered in a single dosage form or divided into several, such as two, three or four dosage forms.

[0047] The present invention also provides use of the above-mentioned pharmaceutical composition in preventing and treating HIV infection.

[0048] The drug of the present invention can be used directly alone for the treatment and prevention of HIV infection, or it can be used in combination with one or more other antiviral drugs to achieve the purpose of improving the overall therapeutic 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-mentioned reverse transcriptase inhibitors can be one or more of AZT, 3TC, ddI, d4T, ddT, TDF, Abacavir, Nevirapine, Efavirenz, Delavirdine, Azvudine, Ainovirine, etc.; the above-mentioned protease inhibitors can be one or more of Saquinavir mesylate, Idinavir, Ritonavir, Amprenavir, Kaletra and Nelfinavirmesylate; the above-mentioned invasion inhibitors can be one or more of Maraviroc, TAK-779, T20, T2635, Sifuvirtide, Abovirtide, etc.; the above-mentioned integration inhibitors can be one or more of Raltegravir, Dolutegravir and Elvitegravi, etc.

[0049] For any particular patient, the specific therapeutically effective dosage level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific active ingredient being employed; the specific composition being employed; the patient's age, weight, general health, sex and diet; the time of administration, route of administration and excretion rate of the specific active ingredient being employed; the duration of treatment; drugs used in combination or concomitantly with the specific active ingredient being employed; and similar factors well known in the medical arts. For example, it is practice in the art to start the dose of the active ingredient at a level lower than that required to obtain the desired therapeutic effect and gradually increase the dose until the desired effect is achieved.

[0050] The beneficial effects of the present invention are:

[0051] The lipopeptide or its pharmaceutically acceptable salt provided by the present invention has stable properties and is a highly effective, novel anti-HIV membrane fusion inhibitor with higher antiviral activity. It can be used to prepare a pharmaceutical composition for preventing and / or treating diseases caused by HIV infection. The pharmaceutical composition is used to prevent and / or treat diseases caused by HIV infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 Anti-HIV-1 activity of newly designed lipopeptide membrane fusion inhibitors;

[0053] Figure 2 The inhibitory activity of LP-37 and LP-39 against different subtypes of HIV-1 strains;

[0054] Figure 3 The inhibitory activity of LP-37 and LP-39 on HIV-1 Env-mediated cell fusion;

[0055] Figure 4 The inhibitory activity of LP-37 and LP-39 against HIV-2 and SIV strains;

[0056] Figure 5 is the inhibitory activity of LP-37 and LP-39 against T20 and HP23 resistant mutants;

[0057] Figure 6 The secondary structure and thermal stability of LP-37 and LP-39;

[0058] Figure 7 These are the results of in vitro metabolic stability analysis of LP-37 and LP-39. DETAILED DESCRIPTION

[0059] The present invention is further described in detail below in conjunction with specific embodiments, and the examples given are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way. Those skilled in the art can refer to the contents of this article and appropriately improve the relevant parameters to achieve. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included within the scope of the present invention. The method of the present invention has been described by preferred embodiments, and relevant personnel can obviously change or appropriately change and combine the compounds and preparation methods described herein without departing from the content, spirit and scope of the present invention to realize and apply the technology of the present invention.

[0060] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0061] Example 1: Design and synthesis of lipopeptides

[0062] LP-25 and LP-26 are lipopeptides modified by stearic acid and cholesterol, respectively, with PEG8 linkers developed by the inventor based on the 2P23 short peptide in the early stage. In order to develop more effective HIV fusion inhibitors, this embodiment continues to design cholesterol-modified lipopeptides with different connecting arms. First, the second amino acid "M" at the N-terminus of LP-26 is replaced with "L", and amino acid sequences of different lengths are introduced at its C-terminus to obtain lipopeptides LP-30, LP-31 and LP-32, respectively. The introduced amino acid sequence may be used as a natural linker between the 2P23 polypeptide sequence and cholesterol. Secondly, lipopeptides LP-33 and LP-34 were obtained by introducing a linker of a common combination of glycine (G) and serine (S). Next, repeated "EAAAK" and "EP" rigid linkers were introduced between the polypeptide sequence and cholesterol to obtain lipopeptides LP-35, LP-36, LP-37, LP-38 and LP-39.

[0063] The lipopeptides involved in this example were synthesized on rinkamide 4-methylphenylhydroxylamine (MBHA) resin using the standard solid phase 9-fluorenylmethoxycarbonyl (FMOC) method. All peptides were acetylated at the N-terminus and aminated at the C-terminus. For cholesterol conjugation, the template peptide contained a lysine residue at the carboxyl terminus with a 1-(4,4-dimethyl-2,6-dioxycyclohexylethylene) ethyl side chain protecting group and a special deprotection step in a 2% hydrazine hydrate-n,n-dimethylformamide (DMF) solution for fatty acid conjugation. Finally, it was purified by reverse phase high performance liquid chromatography (HPLC) with a purity of >95%. The specific peptide names and sequence structures are as follows:

[0064] Lipopeptide name Sequence structure LP-25 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLK-PEG8-K(C18)-NH2]]> LP-26 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLK-PEG8-C(chol)-NH2]]> LP-30 <![CDATA[Ac-ELTWEEWEKKVEELEKKIEELLKKAEEQQKK(chol)-NH2]]> LP-31 <![CDATA[Ac-ELTWEEWEKKVEELEKKIEELLKAAQEQQEK(chol)-NH2]]> LP-32 <![CDATA[Ac-ELTWEEWEKKVEELEKKIEELLKKAEEQQKKNEK(chol)-NH2]]> LP-33 <![CDATA[Ac-ELTWEEWEKKVEELEKKIEELLKGSGGSGGK(chol)-NH2]]> LP-34 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKGGGGSGGGGSK(chol)-NH2]]> LP-35 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKK(chol)-NH2]]> LP-36 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKEAAAKK(chol)-NH2]]> LP-37 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKAEAAAKEAAAKAK(chol)-NH2]]> LP-38 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKEAAAKEAAAK(chol)-NH2]]> LP-39 <![CDATA[Ac-EMTWEEWEKKVEELEKKIEELLKEPEPEPEPEPEPK(chol)-NH2]]>

[0065] Example 2: Discovery and identification of highly active lipopeptides based on rigid linkers

[0066] 1. Experimental Materials and Methods

[0067] The lipopeptides to be tested are LP-30, LP-31, LP-32, LP-33, LP-34, LP-35, LP-36, LP-37, LP-38, LP-39 and control lipopeptides LP-25 and LP-26 synthesized in Example 1. The viruses used are HIV-1 strain NL4-3 and JRFL pseudovirus, prepared by this laboratory; the target cell TZM-bl is a product of the National Laboratory Cell Resource Sharing Service Platform. The specific experimental steps are as follows:

[0068] ① Preparation of HIV-1 pseudovirus: The plasmid expressing the envelope protein (Env) of NL4-3 or JRFL was co-transfected with the HIV-1 backbone plasmid pSG3Δenv into HEK293T cells; the transfected cells were cultured in a cell culture incubator at 37°C and 5% CO2 for 48 hours, and then the supernatant was collected, filtered and the filtrate was collected, which is the pseudovirus liquid containing NL4-3 pseudovirus or JRFL or SF162, and stored at -80°C after titration for future use.

[0069] ② Dissolve the test lipopeptide with deionized water or dimethyl sulfoxide (DMSO), then dilute the lipopeptide to the starting concentration with DMEM medium, and then dilute the lipopeptide 3-fold in a 96-well cell culture plate to obtain a lipopeptide dilution. Set 9 dilutions for each test lipopeptide.

[0070] ③ In a 96-well cell culture plate, add lipopeptide diluent (50 μL / well) to the drug wells and DMEM medium (50 μL / well) to the control wells, and set up 3 replicates. Then add 1000 TCID 50 The virus solution (adjusted to 50 μL / well) was added and incubated at room temperature for 30 minutes.

[0071] ④ Resuspend the pre-cultured target cells TZM-bl in DMEM medium and adjust the cell concentration to 10×10 4 cells / mL, then DEAE-dextran was added to make the final concentration of DEAE-dextran 15μg / mL. Finally, the cells (100μL / well) were added to the 96-well plate containing the virus & lipopeptide complex and cultured in a cell culture incubator at 37°C and 5% CO2 for 48 hours.

[0072] ⑤ Discard the cell culture supernatant, add 30 μL of cell lysis buffer (Promega, Cat. No. E1531) to each well, lyse at room temperature for 15 minutes, then add luciferase detection substrate reagent (Promega, Cat. No. E1501), measure the relative fluorescence unit (RLU) with a microplate photometer, make an inhibition rate curve and calculate the drug half inhibitory concentration (IC 50 ).

[0073] 2. Experimental results and analysis

[0074] The results are as follows Figure 1 As shown, compared with LP-26 with a PEG8 flexible linker, the anti-HIV activity of the lipopeptides LP-30 and LP-31 with 7 extended amino acids "KAEEQQK" at the C-terminus was not improved; while the lipopeptide LP-32 with 10 extended amino acids "KAEEQQKKNE" at the C-terminus inhibited CXCR4-tropic HIV NL4-3strains; in addition, LP-33 with a shorter "GSGGSGG" flexible linker showed lower antiviral activity, while LP-34 with a longer "GGGGSGGGGS" flexible linker showed slightly increased antiviral activity; the above results show that relatively long linkers can improve the antiviral activity of inhibitors. Among the lipopeptide inhibitors with introduced rigid linkers, the antiviral activity of LP-35 with a single "EAAAK" linker was not improved, and LP-36 and LP-37 with two repeated "EAAAK" sequence linkers showed significantly increased antiviral activity. Obviously, the two helix-promoting amino acids (alanine) on both sides of the linker in LP-37 can further improve the inhibitory activity. LP-38 with three repeated "EAAAK" sequence linkers and LP-39 with six repeated "EP" sequence linkers showed antiviral activity comparable to LP-37. The above results show that rigid linkers with repeated "EAAAK" or "EP" amino acid sequences are better than PEG8 or repeated "GGGGS" amino acid sequence linkers; in contrast, LP-37 and LP-39 with 12 amino acids "AEAAAKEAAAKA" and "EPEPEPEPEPEP" rigid linkers showed the greatest optimization and exhibited the highest antiviral activity relative to LP-26 containing a flexible linker. LP-37 and LP-39 inhibited HIV-1 NL4-3 IC 50 The values ​​were 8.08 and 8.17 pM, respectively, inhibiting HIV-1 JRFL IC 50 The values ​​are 76.85 and 56.96 pM respectively.

[0075] Example 3: Broad-spectrum inhibitory activity of LP-37 and LP-39 against HIV-1, HIV-2 and SIV

[0076] AIDS is mainly caused by HIV, which is divided into two types: HIV-1 and HIV-2. HIV-1 is more common, highly contagious, and widely distributed. 95% of infections are caused by HIV-1; HIV-2 is mainly distributed in Africa and shows a local epidemic trend. HIV-1 is highly variable and has produced many subtypes and recombinant viruses in evolution. Among them, HIV-1 subtypes A, B and C are the main viruses that cause the global AIDS epidemic, while in China, A / E and B / C recombinant viruses are the main ones. This example further evaluates the antiviral activity of the above-mentioned superior lipopeptides LP-37 and LP-39 against a group of 36 strains of HIV-1, HIV-2 and SIV through pseudovirus inhibition experiments, virus replication inhibition experiments and fusion inhibitor experiments based on the DSP system.

[0077] 1. Experimental Materials and Methods

[0078] (1) Virus inhibition experiment

[0079] The lipopeptides tested were LP-37 and LP-39 synthesized in Example 1. The HIV-1 pseudovirus used was Figure 2 As shown, the SIV pseudovirus used was SIV 239 and SIV PBJ The HIV-2 replicative virus used was HIV-2 ROD and HIV-2 ST The target cell TZM-bl is a product of the National Laboratory Cell Resource Sharing Service Platform. The specific experimental steps are as follows:

[0080] ① Virus preparation: co-transfect HEK293T cells with a plasmid expressing HIV-1 or SIV envelope protein (Env) and HIV-1 backbone plasmid pSG3Δenv; transfect HEK293T cells with a molecular clone plasmid encoding HIV-2 virus strain ROD or ST; culture the transfected cells in a 37°C, 5% CO2 cell culture incubator for 48 hours, collect the supernatant, filter and collect the filtrate, which is the virus solution, and store it at -80°C after titration for later use.

[0081] After obtaining the virus solution, the experimental steps are the same as steps ②-⑤ in Example 2.

[0082] (2) Fusion suppression experiment based on DSP system

[0083] The lipopeptides tested were LP-37 and LP-39 synthesized in Example 1. Plasmids expressing different subtypes of HIV-1 Env were stored in our laboratory; pDSP8-11 fluorescent reporter plasmid and plasmid stably expressing CXCR4 / CCR5 and DSP 8-11 The target cells 293FT were provided by Professor Zene Matsuda of the University of Tokyo, Japan, and are routinely used and stored in our laboratory; 293T cells were purchased from the American Model Culture Collection. The specific experimental steps are as follows:

[0084] ① 293T cells (effector cells) were plated in 96-well plates (about 1.5×10 4 pcs / well), and stably expressed CXCR4 / CCR5 and DSP 8-11 The target cells 293FT (about 1.5×10 4 / mL) were plated in a 10 cm cell culture dish and cultured at 37°C and 5% CO2.

[0085] ② After 16 hours of culture, HIV-1Env expression plasmid and DSP 1-7 The plasmids were co-transfected into 293T effector cells at a 1:1 ratio.

[0086] ③ 24 hours after transfection, dissolve the lipopeptide membrane with deionized water and dilute the inhibitor to the starting concentration with DMEM medium, add it to the effector cells in the 96-well culture plate at a 3-fold dilution ratio (50 μL / well), set up 9 dilution gradients, 3 replicates. Use DMEM medium without inhibitor as a control. Incubate the culture plate in a 37°C, 5% CO2 cell culture incubator for 1 hour.

[0087] ④ Resuspend the 293FT cells and adjust the cell concentration to about 30×10 4 EnduRen living cell substrate (Promega) was added at a ratio of 1:4000, mixed, and incubated at 37°C and 5% CO2 for 30 minutes.

[0088] ⑤ Add 100 μL of 293FT cells to each well of HIV-1 effector cells, centrifuge at 300 g for 1 minute to allow the effector cells and target cells to fully contact, and incubate at 37°C for 1 hour. Then measure the luciferase activity (relative fluorescence unit, RLU), make an inhibition rate curve, and calculate the drug half inhibitory concentration (IC 50 ).

[0089] 2. Experimental results and analysis

[0090] The inventors have previously reported that LP-25 can effectively inhibit a group of different subtypes of HIV-1 pseudoviruses. This example uses the same group of HIV-1 pseudoviruses to evaluate the antiviral activity of LP-37 and LP-39 in a single-cycle infection assay of TZM-bl cells. Figure 2 As shown in the figure, consistent with previous results, LP-25 effectively inhibited 36 HIV-1 pseudoviruses with an average IC 50 The value was 196.89 pM; the antiviral activity of LP-37 and LP-39 was significantly improved, with an average IC 50 The values ​​are 30.72 and 29.86 pM respectively.

[0091] The inhibitory activities of LP-37 and LP-39 on cell-cell fusion mediated by 7 representative HIV-1 Envs are shown in Figure 2 Figure 3 As shown, the average IC 50 The values ​​were 74.59 and 72.09 pM, respectively, indicating that LP-37 and LP-39 can also effectively inhibit HIV-1 Env-mediated cell-to-cell fusion.

[0092] The inhibitory activity of LP-37 and LP-39 against HIV-2 and SIV strains Figure 4 As shown, LP-37 and LP-39 inhibit HIV-2 ROD Infected IC 50 The values ​​were 215.13 and 371.5 pM respectively; inhibiting HIV-2ST Infected IC 50 The values ​​were 89.57 and 92.99 pM, respectively. LP-37 and LP-39 inhibit SIV 239 IC 50 The values ​​were 17.14 and 25.2 pM respectively; inhibiting SIV PBJ IC 50 The values ​​were 38.8 and 41.97 pM.

[0093] Example 4: LP-37 and LP-39 maintain effective inhibitory activity against drug-resistant HIV-1 strains

[0094] Another important biological characteristic of HIV is that it is easy to develop drug resistance, which leads to treatment failure and is a key issue that needs to be addressed in drug development. This example uses two groups of HIV-1 drug-resistant virus strains: the first group is T20 drug-resistant mutant strains carrying single-point mutations or double mutations (I37T, V38A, V38M, Q40H, N43K, G36S / V38M, I37T / N43K, V38A / N42T), which show high resistance to T20; the second group is HP23 drug-resistant mutant strains carrying single-point mutations, double mutations or triple mutations (E49A , E49K, Q52R, L57R, N126K, E136G, N43K / E49A, E49K / N126K, L57R / E136G, Q39R / N43K / N126K, N43K / E49A / N126K), these mutants showed mild or high resistance to 2P23; the antiviral activity of LP-37 and LP-39 was evaluated using the pseudovirus inhibition experiment in the above Example 2.

[0095] The results are as follows Figure 5 As shown in Figure 2, LP-37 and LP-39 showed great inhibitory activity against drug-resistant mutations. Their average IC 50 The mean values ​​were 9.16 and 8.61 pM, respectively, while the IC 50 The values ​​were 7.28 and 7.45 pM, respectively. According to the results of drug resistance multiple calculation, the IC 50 The change multiples were all less than 3 times, indicating that LP-37 and LP-39 were sensitive to all T20 resistance mutations and maintained a high level of activity. As for HP23 resistance mutants, LP-37 and LP-39 showed mild resistance to several mutants and also showed efficient inhibitory activity against other mutants, with an average IC 50The values ​​were 22.13 and 41.91 pM, respectively. Interestingly, LP-37 inhibited the L57R mutant without obvious resistance, while LP-39 showed 6.73-fold resistance to the L57R mutant; LP-37 inhibited the L57R / E136G mutant with a resistance factor of 7.4, while LP-39 showed a resistance factor of 37.72 to the L57R / E136G mutant; LP-37 had a significantly higher activity level than the lipopeptide membrane fusion inhibitors such as LP-19 and LP-25 previously reported by the inventors' team. In contrast, LP-37 was the most effective inhibitor against HP23 resistant mutants.

[0096] Example 5: Structural features of LP-37 and LP-39 and their interactions with target sequences

[0097] 1. Experimental Materials and Methods

[0098] Circular dichroism (CD) spectroscopy was used to determine the secondary structure (α-helix) and thermal stability of the lipopeptide inhibitor and the interaction between the lipopeptide and the target sequence mimicking polypeptide. The target sequence mimicking polypeptide N36 (Ac-SGIVQQQNNLLRAIEAQQHLLQLTVWGIKQLQARIL-NH2) was derived from the NHR sequence of HIV-1gp41. The lipopeptides to be tested were LP-37 and LP-39 synthesized in Example 1, and the test complex was a mixture of the lipopeptide and N36. The specific experimental steps were:

[0099] ① Use phosphate buffered saline (PBS, pH 7.2) to prepare the test peptide (or test complex) into a 10 μM solution (for the test complex, 10 μM means that the concentrations of the test lipopeptide and N36 are both 10 μM), and place it in a 37°C water bath for 30 minutes.

[0100] ② The solution obtained in step 1 was transferred to the corresponding cuvette, and the changes in the molar ellipticity [θ]λ of the solution in the wavelength range of 195-270nm were scanned using a Jasco spectropolarimeter (model J-815). The typical α-helix structure can show maximum negative peaks at 208nm and 222nm. The PBS blank control was subtracted to correct the spectrum value. During the calculation process, the peak value of -33000degree.cm2.dmol-1 was used as the standard for 100% α-helix content, and the percentage of α-helix content was calculated based on the molar ellipticity of the solution at 222nm.

[0101] ③ Add the solution obtained in step 1 to the thermal stability detection cuvette, and adjust the CD temperature control module to scan the solution [θ]222 at 2-98°C at a speed of 2°C per minute. Smooth the melting curve, and use Origin software to calculate the midpoint temperature (Tm) of the thermal dissociation transition to reflect the thermal stability of the helix.

[0102] 2. Experimental results and analysis

[0103] The results are as follows Figure 6 As shown in A, LP-37 and LP-39 alone showed typical double negative peaks at 208 and 222 nm, indicating that LP-37 and LP-39 alone formed typical α-helical structures with α-helical contents of 96% and 44%, respectively; however, the melting curve could not accurately calculate its Tm value, but the thermal stability of LP-37 was significantly higher than that of LP-39. In contrast, LP-37 with a rigid linker with a repeated "EAAAK" sequence significantly increased the α-helical structure and thermal stability of the lipopeptide. The relatively high self-α-helical content can effectively reduce the binding entropy of the lipopeptide with the target, increase the binding constant, and improve the activity; the relatively stable secondary structure can also improve the stability of the lipopeptide in vivo, which is beneficial to the antiviral activity.

[0104] like Figure 6 As shown in Figure B, both LP-37 and LP-39 can interact with the target sequence mimic peptide N36 to form a stable α-helical structure; the α-helical content of the N36 / LP-37 complex is 93%, and the Tm value is 93°C; the α-helical content of the N36 / LP-39 complex is 65%, and the Tm value is 88°C. In contrast, LP-37 with a rigid linker with a repeated "EAAAK" sequence significantly enhances its ability to bind to the viral target sequence while maintaining stable binding to the target.

[0105] Example 6: Stability study of LP-37 and LP-39

[0106] In order to further verify the drugability of the membrane fusion inhibitor disclosed in the present invention, in this example, LP-37 and LP-39 were placed at 4°C or 37°C for a long time, incubated with human serum, and digested with protease, and then their stability was determined by detecting changes in the antiviral activity of the lipopeptides.

[0107] 1. Experimental Materials and Methods

[0108] Temperature stability experiment: The lipopeptide was prepared into an aqueous solution with a concentration of 500 μM and stored at 4°C and 37°C for 0, 3, 7, 14, 21, 28, 35, 49 or 63 days. The samples were collected and diluted to the IC value of the corresponding lipopeptide. 90 The concentration (i.e., the concentration at which the lipopeptide achieves 90% inhibition of the virus) was used to detect the effects of different samples at this concentration on HIV-1 using the pseudovirus inhibition experiment in Example 2. NL4-3 The inhibitory effect on TZM-bl cells was measured and the inhibition rate was calculated.

[0109] Human serum stability test: The lipopeptide was prepared into an aqueous solution with a concentration of 500 μM, and human serum with a final concentration of 20% was added thereto. After mixing, the mixture was incubated at 37°C for 0, 5, 30, 60, 120, 180 or 240 min, and the samples were collected and diluted to the IC value of the corresponding lipopeptide. 90 The concentration (i.e., the concentration at which the lipopeptide achieves 90% inhibition of the virus) was used to detect the effects of different samples at this concentration on HIV-1 using the pseudovirus inhibition experiment in Example 2. NL4-3 The inhibitory effect on TZM-bl cells was measured and the inhibition rate was calculated.

[0110] Protease digestion: LP-127 or LP-128 was mixed with proteinase K, trypsin, or α-chymotrypsin (Sigma-Aldrich products, catalog numbers P2308, T4799, and C4129, respectively) at a ratio of 20:1 (final concentrations of 2 mg / mL and 0.1 mg / mL, respectively). The samples were collected after incubation at 37°C for 0, 30, 60, 120, 180, or 240 minutes and diluted to the IC value of the corresponding lipopeptide. 90 The concentration (i.e., the concentration at which the lipopeptide achieves 90% inhibition of the virus) was used to detect the effects of different samples at this concentration on HIV-1 using the pseudovirus inhibition experiment in Example 2. NL4-3 The inhibitory effect on TZM-bl cells was measured and the inhibition rate was calculated.

[0111] Protease digestion: Lipopeptides were mixed with proteinase K, trypsin, or α-chymotrypsin (Sigma-Aldrich products, catalog numbers P2308, T4799, and C4129, respectively) at a ratio of 20:1 (final concentrations were 2 mg / mL and 0.1 mg / mL, respectively). Samples were collected after incubation at 37°C for 0, 30, 60, 120, 180, or 240 min and diluted to the IC value of the corresponding lipopeptide. 90 The concentration (i.e., the concentration at which the lipopeptide achieves 90% inhibition of the virus) was used to detect the effects of different samples at this concentration on HIV-1 using the pseudovirus inhibition experiment in Example 2. NL4-3 The inhibitory effect on TZM-bl cells was measured and the inhibition rate was calculated.

[0112] 2. Experimental results and analysis

[0113] like Figure 7As shown in the figure, at 4°C, LP-37 was highly stable, and its antiviral activity remained basically unchanged after 63 days; the activity of LP-39 showed a downward trend, and its activity remained at about 77% of the original level after 63 days. At 37°C, LP-37 was still highly stable and maintained its original antiviral activity after 49 days, and its activity remained at about 80% of the original level after 63 days; while LP-39 showed a significant decrease in activity, and almost lost its inhibitory activity after 49 days. In contrast, LP-37 with a rigid linker with a repeated "EAAAK" sequence was more stable than LP-39 in terms of temperature. After treatment with proteinase K, trypsin, and α-chymotrypsin, LP-37 was highly resistant to digestion by all three proteases, and its antiviral activity did not change significantly; while LP-39 was relatively stable in trypsin and α-chymotrypsin, but LP-39 was very sensitive to proteinase K, and its antiviral activity decreased significantly after proteinase K treatment. In addition, both lipopeptides maintained stable antiviral activity after incubation with human serum samples at 37°C. In contrast, LP-37 with a rigid linker with a repetitive "EAAAK" sequence showed obvious superiority in stability.

[0114] The present disclosure is described in detail above. For those skilled in the art, the present disclosure can be implemented in a wider range under equivalent parameters, concentrations and conditions without departing from the purpose and scope of the present disclosure and without unnecessary experiments. Although the present disclosure provides specific embodiments, it should be understood that further improvements can be made to the present disclosure. In short, according to the principles of the present disclosure, this application intends to include any changes, uses or improvements to the present disclosure, including changes that are out of the scope of the disclosure in the present application and are made with conventional techniques known in the art. Some basic features can be applied within the scope of the following claims.

Claims

1. A lipopeptide or a pharmaceutically acceptable salt thereof, characterized in that: The lipopeptide is shown in the following formula I or formula II: Formula I: X1-EMTWEEWEKKVEELEKKIEELLK-X2-X3-X4-X5; Formula II: X1-ELTWEEWEKKVEELEKKIEELLK-X6-X7-X4-X5; Wherein, X1 is an amino terminal protecting group, X4 is a lipophilic compound group used to modify the C-terminus, and X5 is a carboxyl terminal protecting group; The X2 and X3 are any one of the following 1) and 2): 1) X2 is a polypeptide sequence (EAAAK)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine, cysteine, 2,3-diaminopropionic acid, ornithine, 2,4-diaminobutyric acid, 2,7-diaminoheptanoic acid or does not exist; 2) X2 is a polypeptide sequence A[(EAAAK)n]A or (EP)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine, cysteine, 2,3-diaminopropionic acid, ornithine, 2,4-diaminobutyric acid or 2,7-diaminoheptanoic acid; The X6 is a polypeptide sequence KAEEQQKKNE, and the X7 is lysine, cysteine, 2,3-diaminopropionic acid, ornithine, 2,4-diaminobutyric acid or 2,7-diaminoheptanoic acid.

2. The lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The amino terminal protecting group is any one of acetyl, amino, maleyl, succinyl, tert-butyloxycarbonyl or benzyloxy or other hydrophobic groups or macromolecular carrier groups; And / or, the carboxyl terminal protecting group is any one of amino, carboxyl, hydroxyl, amide or tert-butyloxycarbonyl or other hydrophobic groups or macromolecular carrier groups.

3. The lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The amino terminal protecting group is an acetyl group; and / or the carboxyl terminal protecting group is an amino group.

4. The lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The lipophilic compound is any one of cholesterol succinate, 2-cholesterol acetic acid, 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, cholesteryl bromoacetate, fatty acids containing 8 to 20 carbon atoms, dihydrosphingosine, and vitamin E; Preferably, the lipophilic compound is cholesterol succinate.

5. The lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The X2 and X3 are any one of the following 1) and 2): 1) X2 is a polypeptide sequence (EAAAK)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine or does not exist; 2) X2 is a polypeptide sequence A[(EAAAK)n]A or (EP)n, wherein n represents the number of repetitions, and n is 1, 2, 3, 4, 5 or 6; X3 is lysine; The X7 is lysine.

6. The lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The lipopeptide is one of the compounds shown in the following formula: Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKK(chol)-NH2; Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKEAAAKK(chol)-NH2; Ac-EMTWEEWEKKVEELEKKIEELLKAEAAAKEAAAKAK(chol)-NH2; Ac-EMTWEEWEKKVEELEKKIEELLKEAAAKEAAAKEAAAK(chol)-NH2; Ac-EMTWEEWEKKVEELEKKIEELLKEPEPEPEPEPK(chol)-NH2; Ac-ELTWEEWEKKVEELEKKIEELLKKAEEQQKKNEK(chol)-NH2.

7. A pharmaceutical composition, characterized in that It comprises the lipopeptide according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

8. Use of the lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, or the pharmaceutical composition according to claim 7, in the preparation of a lipopeptide HIV membrane fusion inhibitor.

9. Use of the lipopeptide or a pharmaceutically acceptable salt thereof according to claim 1, or the pharmaceutical composition according to claim 7 in the preparation of any of the following drugs: 1) Antiviral drugs; 2) Drugs for preventing and / or treating diseases caused by viral infections; 3) Drugs that inhibit virus-cell fusion; 4) Drugs that inhibit viruses from invading cells; 5) Drugs that inhibit viral replication; in, The virus is one or more of HIV1, HIV-2 and SIV.

10. The use according to claim 9, characterized in that The disease caused by the viral infection is AIDS.

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