WO2013075594A1 - Artificially designed anti-hiv infection polypeptide, composition and use - Google Patents

Artificially designed anti-hiv infection polypeptide, composition and use Download PDF

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WO2013075594A1
WO2013075594A1 PCT/CN2012/084433 CN2012084433W WO2013075594A1 WO 2013075594 A1 WO2013075594 A1 WO 2013075594A1 CN 2012084433 W CN2012084433 W CN 2012084433W WO 2013075594 A1 WO2013075594 A1 WO 2013075594A1
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seq
polypeptide
formula
stereoisomer
seelikk
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PCT/CN2012/084433
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French (fr)
Chinese (zh)
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刘克良
史卫国
王潮
蔡利锋
贾启燕
王昆
郑保华
张沙
白玉
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中国人民解放军军事医学科学院毒物药物研究所
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Publication of WO2013075594A1 publication Critical patent/WO2013075594A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV

Definitions

  • the invention belongs to the field of biomedicine and relates to an artificially designed, non-natural sequence anti-HIV infection polypeptide, in particular to a polypeptide represented by formula I, a derivative thereof, a stereoisomer thereof, or a physiologically inactive thereof salt.
  • the present invention also relates to a pharmaceutical composition comprising the above-described polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, and a polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or The use of non-physiological salts in the treatment or prevention of diseases associated with HIV infection, especially acquired immunodeficiency syndrome (AIDS).
  • AIDS acquired immunodeficiency syndrome
  • HIV-1 Human immunodeficiency virus type 1
  • HIV fus ion inhibitors are a new class of anti-HIV drugs that interfere with the entry of viruses into target cells. Unlike traditional enzyme inhibitors, they act outside the cell and cut off the spread of the virus at the initial stage of infection. Prevention and control of HIV-1 infection has special significance, and thus has become a hot spot for new mechanisms of anti-HIV drug research.
  • HIV fusion inhibitors target the HIV-1 fusion protein gp41, a functional protein that mediates fusion of HIV-1 to target cell membranes. HIV fusion inhibitors inhibit the formation of core hexamer structures (6-HB) by membrane fusion by inhibiting gp41, thereby inhibiting virus entry into target cells.
  • T-20 is the first and currently marketed HIV-1 fusion inhibitor, a native sequence 36 peptide (referred to as C-peptide) derived from the HIV-1 gp41 CHR domain, which was approved by the FDA in 2003.
  • T20 binds to gp41, which competitively inhibits the formation of functional core structures mediated by gp41 and blocks the fusion of HIV-1 with target cell membranes. Inhibition of viral infection of target cells.
  • T20 Although the discovery of T-20 opened up a new field of using peptide drugs to control HIV, its limitations and limitations have limited the widespread use of T20.
  • the first is drug resistance. Since T20 is completely derived from the gp41 native sequence, it is highly sensitive to target mutations. A single amino acid residue mutation in the target sequence results in a 10-fold decrease in T20 activity, and two residue mutations reduce its activity. 100 times. Therefore, although the native sequence of T20 retains the structure of the natural ligand to the utmost extent, it is also susceptible to T20 resistance due to the high mutation of the target HIV-1 gp41 sequence. Secondly, T20 is easily degraded by proteases and has low bioavailability in vivo. Therefore, how to overcome drug resistance and improve the stability of enzymatic hydrolysis is the main direction of research on a new generation of HIV fusion inhibitors.
  • HIV-1 peptide fusion inhibitors are based primarily on the optimization and modification of active natural ligand sequences.
  • the first generation fusion inhibitors T-20, C34, etc. are the natural sequences directly derived from gp41 CHR; the second generation T1249, Sifuvirtide, T1144, etc. are based on the natural sequence, and the non-conservative amino acid residues The base is obtained by deleting and replacing. Due to the high sensitivity of natural origin sequences to drug resistance mutations, new design ideas need to be explored.
  • the present inventors have proposed an anti-HIV active peptide design idea based on a non-naturally occurring sequence of a target structure: based on the three-dimensional crystal structure of the target gp41 NHR spiral trimer, a computer-aided rational design method is used to design a non-natural sequence oc spiral from the beginning.
  • the peptide which mimics the active conformation of the native C-peptide, binds to the target to produce the corresponding biological function, thereby designing a novel structure of the anti-HIV active polypeptide.
  • the use of non-native sequences and natural sequences is completely non-homologous, exploring new ideas for inhibiting drug resistance. The present invention has thus been completed. Summary of the invention
  • the object of the present invention is to artificially design a new completely non-native sequence of an anti-HIV-infected polypeptide according to the target structure, to achieve an HIV infection-inhibiting activity comparable to or higher than the native sequence, and to inhibit the T20-resistant HIV strain.
  • One aspect of the invention relates to a polypeptide of formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, XalAAXdlXelBB- X a 2 AXd2Xe2BB ⁇ XasAAXdsXesBB- X a 4 AXd4Xe4BB ⁇ XasAAXdsXesBB
  • X al, X a2, X a3 , X a4, X a5 each independently a D or L natural or non-natural hydrophobic or hydrophilic amino acid;
  • A is a D or L natural or non-natural acidic amino acid
  • X dl , X d2 , X d3 , X d4 , X d5 are each independently a D- or L-type natural or non-natural hydrophobic amino acid;
  • Xei. X e2 , X , X e X e5 are each independently a natural or non-natural hydrophobic acid of type D or L;
  • X is a natural or non-natural basic amino acid of the D or L form; as used herein, the term “stereoisomer” refers to the D- or L-stereo configuration of a polypeptide of Formula I.
  • X isoleucine (I ), cineine (V), norleucine (N le ), leucine (L), alanine (A), Serine (S), threonine (T), asparagine (N), and glutamine (Q);
  • A is selected from D-type or L-form glutamic acid (E) or aspartic acid (D);
  • X dl , X d2 , X d3 , X d4 , X d5 are each independently selected from the following amino acids of type D or L: glutamine (Q), tryptophan (W), naphthyl alanine (N al ), 3-quinoline alanine (Q al ), isoleucine (I ), cineine (V), norleucine, leucine (L), alanine (A), asparagine ( N), serine (S), threonine (T), and tyrosine (Y);
  • Xei. X e2 , X , X , X e5 are each independently selected from the following amino acids of type D or L: isoleucine (I ), cineine (V), norleucine, leucine (L) , alanine (A), serine (S), glutamine (Q), asparagine (N), threonine (T), and tyrosine (Y);
  • polypeptide of Formula I is selected from lysine (K) or arginine (R) of type D or type L.
  • polypeptide of Formula I a derivative thereof, a stereoisomer thereof, or a salt thereof that is not physiologically toxic, wherein the polypeptide of Formula I is selected from the group consisting of:
  • N al EEN al N IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 10)
  • NEEQIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 68); another aspect of the invention relates to a pharmaceutical composition comprising at least one of the above-described polypeptides of formula I, derivatives thereof, A construct, or a salt thereof that is not physiologically toxic, and a pharmaceutically acceptable carrier or adjuvant.
  • a further aspect of the invention relates to an HIV fusion inhibitor comprising at least one of the above-described polypeptides of formula I, derivatives thereof, stereoisomers thereof, or salts thereof which are not physiologically toxic.
  • a further aspect of the invention relates to the use of a polypeptide of the above formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, for the preparation of an HIV fusion inhibitor.
  • a further aspect of the invention relates to the use of a polypeptide of the above formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic for the preparation of a medicament for the treatment or prevention of a disease associated with HIV infection, in particular AIDS.
  • a further aspect of the invention relates to a method of treating or preventing a disease associated with HIV infection, in particular AIDS, comprising administering an effective amount of a polypeptide of formula I, a derivative thereof, or a stereoisomer thereof to a subject to be treated or prevented. Body, or a salt thereof that is not physiologically toxic.
  • the invention also relates to a method of using the polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, comprising an effective amount of the polypeptide of the formula I, a derivative thereof The stereoisomer, or a salt thereof that is not physiologically toxic, is administered to a subject in need thereof.
  • the term "natural” refers to a naturally occurring, unmodified, such as the amino acid sequence of HIV-1 gp41 itself.
  • non-native means artificially designed, different from nature, such as modified, modified sequences of directly derived natural sequences, and the like.
  • hydrophobic amino acid means an amino acid whose side chain is a hydrophobic group, and includes: Ala, Val, Leu, He, Met, Phe, Trp and the like.
  • hydrophilic amino acid means an acid having a hydrophilic group capable of forming a hydrogen bond with water, and includes: Ser, Thr, Cys, Asp, Asn, Glu, Gin, Arg , Lys, His, Tyr, etc.
  • the term "acidic amino acid” means an amino acid having a gas group in a side chain, and includes: Glu, Asp and the like.
  • basic acid means an acid having an amino group or a mercapto group in a side chain, and includes: Lys, Arg and the like.
  • Env envelope glycoprotein
  • HOBt 1-Hydroxylbenzotriazole anhydrous
  • NHR N-terminal heptad repeat
  • HIV Human Immunodeficiency Virus
  • HIV-1 Human Immunodeficiency Virus Type I
  • Trp Tryptophan
  • Rink Amide resin used in the DIEA N,N-diisopropylethylamine example is the product of Tianjin Nankai Synthetic Co., Ltd.; the natural amino acid protected by HBTU, H0BT, DIEA and Fmoc is Shanghai Jill Biochemistry. The company and Chengdu Chengnuo New Technology Co., Ltd.
  • Fmoc-protected naphthylalanine and quinoline alanine are laboratory synthetic products; N-methylpyrrolidone (leg P) is ACR0S company product; trifluoroacetic acid ( TFA) is a product of Beijing Bomaijie Technology Co., Ltd.; DMF and DCM are products of South Korea's Samsung; chromatographic pure acetonitrile is Fisher's product. Other reagents are domestically produced pure products if they are not described.
  • a standard Fmoc solid phase peptide synthesis method was employed. All peptide sequences were amidated at the C-terminus and acetylated at the N-terminus.
  • Rink Amide resin is used, the amino acid is in the L configuration, and the peptide chain is extended from the C terminal to the N terminal.
  • the activator is a DMF solution containing HBTU and HOBt, and the activated base is a leg P solution of DIEA.
  • the deprotecting agent is a DMF solution of piperidine.
  • the cleavage agent is trifluoroacetic acid (TFA), and the crude peptide is dissolved in water and stored by lyophilization.
  • the peptide was synthesized using a CEM fully automated microwave peptide synthesizer.
  • the synthesis conditions were as follows: J ⁇ acid: 0. 2M DMF solution,
  • Activator 0. 45M HBTU/HOBt DMF solution (where the molar ratio of HBTU to HOBt is 1: 1),
  • Blocking reagent 20% v/v DMF solution of acetic anhydride.
  • Rected Rink Amide resin 0. 5g (0. 25mmol) was placed in the CEM microwave peptide synthesizer reactor, and then the amino acid, activator, activated base, deprotection reagent, blocking reagent were arranged at the above concentration, and then CEM microwave was used. Automated peptide synthesizer for synthesis. After completion, the peptide resin was washed with DMF for 3 times, then shrunk with anhydrous methanol, and dried under vacuum at room temperature to obtain a peptide resin of 2.05 g.
  • Pyrolysis of Peptide Resin Weigh the peptide resin synthesized by the above microwave synthesizer 2. 05g, put it into a 250ml eggplant-shaped bottle, water bath, and electromagnetic stirring. The lysate was prepared by adding 1 gram of the peptide resin to 10 ml. TFA needs to be cooled in a water bath for 30 minutes or pre-stored in a water tank; the prepared lysate is added to the peptide resin under water bath conditions, electromagnetic stirring, the resin turns orange-red, and the reaction is carried out in a water bath for 30 minutes, then, the water bath is removed, room temperature The reaction was further stirred for 90 min and the reaction was completed.
  • the resulting crude peptide was purified by medium or high pressure preparative color transfer.
  • the color column is a C8 column, and the eluent is acetonitrile, water and a small amount of trifluoroacetic acid.
  • the color column was previously equilibrated with 200 ml of an aqueous solution containing 15% (v/v) acetonitrile and 0.1% (v/v) trifluoroacetic acid solution.
  • the mixture was washed with 200 ml of an aqueous solution containing 15% (v/v) acetonitrile and 0.1% (v/v) trifluoroacetic acid in 0-15 minutes, and the fraction of the eluate was detected by high performance liquid chromatography. 1% ⁇ / ⁇ The acetonitrile content of the eluate was increased to 65% (v / v) in 10-30 minutes (the content of trifluoroacetic acid was unchanged, maintaining 0. 1% ⁇ / ⁇ ) until the main peak of the purified polypeptide is eluted. The eluates were combined, and the organic solvent was removed by rotary evaporation, and the peptide was freeze-dried to obtain a pure peptide content of more than 95% by HPLC.
  • Example 69 Inhibition of HIV-1 biological activity assay
  • HIV-1 chronically infected with H9 cells.
  • H9 cells the HIV-1 virus gene is integrated into the DNA strand of H9 cells and expressed together with H9 cells:
  • the viral membrane protein GP160 is expressed in H9 cells. surface.
  • H9 cells become a huge HIV-1 virus.
  • the whole process can accurately reflect the fusion process of HIV-1 virus particles with target cells.
  • the test compound polypeptide inhibits fusion of HIV-1 infected H9 cells with MT-2 target cells, thereby judging the anti-fusion activity of the test compound.
  • HIV-1 IIIB- infected H9 cells (H9/HIV-1 IIIB , prepared by the laboratory, see Jiang, S., Radigan, L for specific methods) . and Zhang, L. (2000) Proc. SPIE 3926, 212-219) were labeled with a fluorescent reagent, Calcein_AM (Molecular Probes, Inc., Eugene, OR), and then 50 ⁇ l per well was added to a 96-well plate. Labeled H9/HIV-l mB cells (2 X10 5 cells/ml), and then add 50 ⁇ l of different concentrations of test samples (test samples are examples)
  • test compounds Compounds 1-68 of SEQ ID NOS: 1-68 prepared in Examples 1-68) all showed high inhibition of HIV-1 mediated cell fusion activity.

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Abstract

Provided in the present invention are a polypeptide and the derivative and stereoisomer or salt without physiological toxicity thereof as shown in formula I. The polypeptide is an artificially designed anti-HIV infection polypeptide. Also provided in the present invention are pharmaceutical compositions containing the polypeptide and the derivative and stereoisomer or salt without physiological toxicity thereof, and the use of the polypeptide and the derivative and stereoisomer or salt without physiological toxicity thereof in treating or preventing related diseases caused by the HIV infection.

Description

人工设计的抗 H I V感染多肽、 组合物以及用途 技术领域  Artificially designed anti-H I V infectious polypeptide, composition and use thereof
本发明属于生物医药领域， 涉及一种人工设计的、 非天然序列的 抗 HIV感染多肽， 具体地， 涉及式 I所示的多肽、 其衍生物、 其立体 异构体、 或其无生理毒性的盐。 本发明还涉及含有上述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐的药物组合物， 以及 式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐在治疗或 预防 HIV感染所致相关疾病尤其是获得性免疫缺陷综合征 ( AIDS, 即 艾滋病） 的用途。  The invention belongs to the field of biomedicine and relates to an artificially designed, non-natural sequence anti-HIV infection polypeptide, in particular to a polypeptide represented by formula I, a derivative thereof, a stereoisomer thereof, or a physiologically inactive thereof salt. The present invention also relates to a pharmaceutical composition comprising the above-described polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, and a polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or The use of non-physiological salts in the treatment or prevention of diseases associated with HIV infection, especially acquired immunodeficiency syndrome (AIDS).
XalAAXdlXelBB - Xa2AAXd2Xe2BB - XasAAXdsXesBB - X"AAXd4Xe4BB - XasAAXdsXesBB XalAAXdlXelBB - X a 2AAXd2Xe2BB - XasAAXdsXesBB - X"AAXd4Xe4BB - XasAAXdsXesBB
式 I 背景技木  Formula I background
艾滋病目前在世界范围内流行， 给人类健康带来严重威胁。 人免 疫缺陷病毒 I型 （ HIV- 1 )是造成 AIDS感染的主要病原体。 由于传统 的逆转录酶抑制剂及蛋白酶抑制剂耐药性问题日益突出， 使得开发作 用于 HIV- 1 生命周期新靶点药物成为共识。 HIV 融合抑制剂 （HIV fus ion inhibitors )是干扰病毒进入靶细胞的新一类抗 HIV药物， 与传统的酶抑制剂不同， 它作用于细胞外， 在感染的初始环节切断病 毒的传播， 这对于预防及控制 HIV-1感染具有特殊意义， 因而成为新 机制抗 HIV药物研究的热点。  AIDS is currently prevalent worldwide, posing a serious threat to human health. Human immunodeficiency virus type 1 (HIV-1) is the main cause of AIDS infection. As the traditional reverse transcriptase inhibitors and protease inhibitors become more and more prominent, the development of new targets for the HIV-1 life cycle has become a consensus. HIV fus ion inhibitors are a new class of anti-HIV drugs that interfere with the entry of viruses into target cells. Unlike traditional enzyme inhibitors, they act outside the cell and cut off the spread of the virus at the initial stage of infection. Prevention and control of HIV-1 infection has special significance, and thus has become a hot spot for new mechanisms of anti-HIV drug research.
HIV融合抑制剂以 HIV- 1融合蛋白 gp41为作用靶标， gp41是介 导 HIV- 1 与靶细胞膜融合的功能蛋白质。 HIV 融合抑制剂通过抑制 gp41促进膜融合的核心六聚体结构（ 6- HB )的形成， 从而抑制病毒进 入靶细胞。 T- 20是第一个也是目前唯一上市的 HIV- 1融合抑制剂， 它 是衍生于 HIV- 1 gp41 CHR功能区的天然序列 36肽（称为 C肽） ， 于 2003年被 FDA批准。 T20能够与 gp41结合， 从而竟争性抑制 gp41介 导融合的功能性核心结构形成， 阻断 HIV- 1与靶细胞膜的融合过程， 抑制病毒感染靶细胞。 HIV fusion inhibitors target the HIV-1 fusion protein gp41, a functional protein that mediates fusion of HIV-1 to target cell membranes. HIV fusion inhibitors inhibit the formation of core hexamer structures (6-HB) by membrane fusion by inhibiting gp41, thereby inhibiting virus entry into target cells. T-20 is the first and currently marketed HIV-1 fusion inhibitor, a native sequence 36 peptide (referred to as C-peptide) derived from the HIV-1 gp41 CHR domain, which was approved by the FDA in 2003. T20 binds to gp41, which competitively inhibits the formation of functional core structures mediated by gp41 and blocks the fusion of HIV-1 with target cell membranes. Inhibition of viral infection of target cells.
尽管 T- 20的发现开辟了利用肽类药物控制 HIV的新领域， 但由 于其本身存在的一些缺陷和不足， 限制了 T20的广泛应用。 首先是耐 药性， 由于 T20完全衍生于 gp41天然序列， 对靶标突变的敏感性高， 靶标序列单个氨基酸残基突变即会导致 T20活性降低 10倍， 两个残 基突变则会使其活性降低 100倍。 因此， T20的天然序列虽然最大限 度地保持了天然配基的结构， 但亦由于靶标 HIV-1 gp41序列的高突 变性而导致病毒容易对 T20产生耐药性。其次， T20易被蛋白酶降解， 体内生物利用度低。 因此， 如何克服耐药性以及提高酶解稳定性， 是 新一代 HIV融合抑制剂研究的主要方向。  Although the discovery of T-20 opened up a new field of using peptide drugs to control HIV, its limitations and limitations have limited the widespread use of T20. The first is drug resistance. Since T20 is completely derived from the gp41 native sequence, it is highly sensitive to target mutations. A single amino acid residue mutation in the target sequence results in a 10-fold decrease in T20 activity, and two residue mutations reduce its activity. 100 times. Therefore, although the native sequence of T20 retains the structure of the natural ligand to the utmost extent, it is also susceptible to T20 resistance due to the high mutation of the target HIV-1 gp41 sequence. Secondly, T20 is easily degraded by proteases and has low bioavailability in vivo. Therefore, how to overcome drug resistance and improve the stability of enzymatic hydrolysis is the main direction of research on a new generation of HIV fusion inhibitors.
目前的 HIV- 1肽类融合抑制剂， 主要是基于活性天然配基序列的 优化及改造。如第一代融合抑制剂 T- 20，C34等是直接衍生于 gp41 CHR 的天然序列； 第二代 T1249、 西夫韦肽、 T1144等则是在天然序列的 基础上， 对其中非保守氨基酸残基进行删除替换而得到的。 由于天然 起源序列对耐药性突变的高敏感性， 需要探索新的设计思路。  Current HIV-1 peptide fusion inhibitors are based primarily on the optimization and modification of active natural ligand sequences. For example, the first generation fusion inhibitors T-20, C34, etc. are the natural sequences directly derived from gp41 CHR; the second generation T1249, Sifuvirtide, T1144, etc. are based on the natural sequence, and the non-conservative amino acid residues The base is obtained by deleting and replacing. Due to the high sensitivity of natural origin sequences to drug resistance mutations, new design ideas need to be explored.
本发明人提出了基于靶标结构的非天然起源序列的抗 HIV活性肽 设计思想： 基于靶标 gp41 NHR螺旋三聚体的三维晶体结构， 应用计 算机辅助的理性设计方法， 从头设计非天然序列的 oc螺旋肽， 模拟天 然 C肽的活性构象， 与靶标结合而产生相应的生物学功能， 由此设计 全新结构的抗 H I V活性多肽。 利用非天然序列与天然序列完全非同源 性特点， 探索抑制耐药性的新思路。 由此而完成了本发明内容。 发明内容  The present inventors have proposed an anti-HIV active peptide design idea based on a non-naturally occurring sequence of a target structure: based on the three-dimensional crystal structure of the target gp41 NHR spiral trimer, a computer-aided rational design method is used to design a non-natural sequence oc spiral from the beginning. The peptide, which mimics the active conformation of the native C-peptide, binds to the target to produce the corresponding biological function, thereby designing a novel structure of the anti-HIV active polypeptide. The use of non-native sequences and natural sequences is completely non-homologous, exploring new ideas for inhibiting drug resistance. The present invention has thus been completed. Summary of the invention
本发明目的是根据靶标结构， 人为设计新的完全非天然序列的抗 HIV感染多肽， 达到与天然序列相当或更高的抑制 HIV感染活性， 同 时能够抑制 T20耐药性 HIV毒株。  The object of the present invention is to artificially design a new completely non-native sequence of an anti-HIV-infected polypeptide according to the target structure, to achieve an HIV infection-inhibiting activity comparable to or higher than the native sequence, and to inhibit the T20-resistant HIV strain.
本发明的一个方面涉及式 I所示的多肽、 其衍生物、 其立体异构 体、 或其无生理毒性的盐， XalAAXdlXelBB- Xa2 AXd2Xe2BB~ XasAAXdsXesBB- Xa4 AXd4Xe4BB~ XasAAXdsXesBB One aspect of the invention relates to a polypeptide of formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, XalAAXdlXelBB- X a 2 AXd2Xe2BB~ XasAAXdsXesBB- X a 4 AXd4Xe4BB~ XasAAXdsXesBB
式 I  Formula I
其中，  among them,
Xal、 Xa2、 Xa3、 Xa4 、 Xa5分别独立地为 D型或 L型的天然或非天然 的疏水性或亲水性氨基酸； X al, X a2, X a3 , X a4, X a5 each independently a D or L natural or non-natural hydrophobic or hydrophilic amino acid;
A为 D型或 L型的天然或非天然酸性氨基酸；  A is a D or L natural or non-natural acidic amino acid;
Xdl、 Xd2、 Xd3、 Xd4、 Xd5分别独立地为 D型或 L型的天然或非天然的 疏水性氨基酸； X dl , X d2 , X d3 , X d4 , X d5 are each independently a D- or L-type natural or non-natural hydrophobic amino acid;
Xei. Xe2、 X 、 Xe Xe5分别独立地为 D型或 L型的天然或非天然疏 水性 ^酸； Xei. X e2 , X , X e X e5 are each independently a natural or non-natural hydrophobic acid of type D or L;
B为 D型或 L型的天然或非天然碱性氨基酸； 本发明所用术语"立体异构体"是指式 I多肽的 D-或 L-立体构型。 在本发明的一个实施方案中， X"、 Xa2、 Xa3、 Xa4 、 Xa5分别独立地 选自 D型或 L型的如下氨基酸： 色氨酸（W) 、 萘丙氨酸（Nal) 、 3- 喹啉丙氨酸（Qal)、 异亮氨酸（I )、 翔氨酸（V)、 正亮氨酸（Nle)、 亮氨酸（L) 、 丙氨酸（A) 、 丝氨酸（S) 、 苏氨酸（T) 、 天冬酰 胺（ N )、 和谷氨酰胺 (Q) ； B is a natural or non-natural basic amino acid of the D or L form; as used herein, the term "stereoisomer" refers to the D- or L-stereo configuration of a polypeptide of Formula I. In one embodiment of the invention, X", Xa2, Xa3, Xa4, Xa5 are each independently selected from the following amino acids of type D or L: tryptophan (W), naphthylalanine (N al ), 3-quinoline alanine (Q al ), isoleucine (I ), cineine (V), norleucine (N le ), leucine (L), alanine (A), Serine (S), threonine (T), asparagine (N), and glutamine (Q);
A选自 D型或 L型的谷氨酸（E)或天冬氨酸（D) ；  A is selected from D-type or L-form glutamic acid (E) or aspartic acid (D);
Xdl、 Xd2、 Xd3、 Xd4、 Xd5分别独立地选自 D型或 L型的如下氨基酸： 谷酰胺（Q)、 色氨酸（W)、 萘丙氨酸（Nal)、 3-喹啉丙氨酸（Qal)、 异亮氨酸（I ) 、 翔氨酸（V) 、 正亮氨酸 、 亮氨酸（L) 、 丙 氨酸（A) 、 天冬酰胺（N) 、 丝氨酸（S) 、 苏氨酸（T) 、 和酪氨酸 (Y) ； X dl , X d2 , X d3 , X d4 , X d5 are each independently selected from the following amino acids of type D or L: glutamine (Q), tryptophan (W), naphthyl alanine (N al ), 3-quinoline alanine (Q al ), isoleucine (I ), cineine (V), norleucine, leucine (L), alanine (A), asparagine ( N), serine (S), threonine (T), and tyrosine (Y);
Xei. Xe2、 X 、 X 、 Xe5分别独立地选自 D型或 L型的如下氨基酸： 异亮氨酸（I ) 、 翔氨酸（V) 、 正亮氨酸 、 亮氨酸（L) 、 丙 氨酸（ A )、丝氨酸（ S )、谷酰胺（ Q )、天冬酰胺（ N )、 苏氨酸（ T )、 和酪氨酸（ Y )； Xei. X e2 , X , X , X e5 are each independently selected from the following amino acids of type D or L: isoleucine (I ), cineine (V), norleucine, leucine (L) , alanine (A), serine (S), glutamine (Q), asparagine (N), threonine (T), and tyrosine (Y);
B选自 D型或 L型的赖氨酸（K)或精氨酸（R) 。 在本发明的一个实施方案中， 所述的式 I多肽、 其衍生物、 其立 体异构体、 或其无生理毒性的盐， 其中， 所述式 I多肽选自如下的化 合物： B is selected from lysine (K) or arginine (R) of type D or type L. In one embodiment of the invention, the polypeptide of Formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof that is not physiologically toxic, wherein the polypeptide of Formula I is selected from the group consisting of:
1. EE D IEELI SEEL IKK IEEQI QEESI K (SEQ ID NO: 1)  1. EE D IEELI SEEL IKK IEEQI QEESI K (SEQ ID NO: 1)
2. EE R IEELI SEEL IKK IEEQI QEESI K (SEQ ID NO: 2)  2. EE R IEELI SEEL IKK IEEQI QEESI K (SEQ ID NO: 2)
3. EE S IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 3)  3. EE S IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 3)
4. EE I IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 4)  4. EE I IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 4)
5. EE A IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 5)  5. EE A IEELIKK SEEL IKK IEEQIKK QEESIKK (SEQ ID NO: 5)
6. NalEENalD IEELIKK SEELI IEEQIKK QEESIKK (SEQ ID NO: 6)； 6. N al EEN al D IEELIKK SEELI IEEQIKK QEESIKK (SEQ ID NO: 6);
7. NalEENalR IEELIKK SEELI IEEQIKK QEESIKK (SEQ ID NO: 7)； 7. N al EEN al R IEELIKK SEELI IEEQIKK QEESIKK (SEQ ID NO: 7);
8. NalEENalS IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 8)； 8. N al EEN al S IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 8);
9. NalEENalQ IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 9)； 9. N al EEN al Q IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 9);
10. NalEENalN IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 10) 10. N al EEN al N IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 10)
11. QalEEQalD IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 11) 11. Q al EEQ al D IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 11)
12. QalEEQalR IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 12) 12. Q al EEQ al R IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 12)
13. QalEEQalS IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 13) 13. Q al EEQ al S IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 13)
14. QalEEQalQ IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 14) 14. Q al EEQ al Q IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 14)
15. QalEEQalN IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 15) 15. Q al EEQ al N IEELIKK SEELIKK IEEQIKK QEESIKK (SEQ ID NO: 15)
16. SEEQIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 16)  16. SEEQIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 16)
17. SEENI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 17)  17. SEENI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 17)
18. SEESIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 18)  18. SEESIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 18)
19. QEESIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 19)  19. QEESIKK WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 19)
20. QEENI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 20)  20. QEENI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 20)
21. NEESI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 21)  21. NEESI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 21)
22. NEEQI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 22)  22. NEEQI WEEWSKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 22)
23. SEEQIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 23)  23. SEEQIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 23)
24. SEENIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 24)  24. SEENIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 24)
25. SEESIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 25) 26. QEESIKK EE D lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 26)25. SEESIKK WEEWDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 25) 26. QEESIKK EE D lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 26)
27. QEENI EE D lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 27)27. QEENI EE D lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 27)
28. NEESI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 28)28. NEESI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 28)
29. NEEQI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 29)29. NEEQI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 29)
30. QEESIKK EE S lEELIKK QEELI IEESI (SEQ ID NO: 30)30. QEESIKK EE S lEELIKK QEELI IEESI (SEQ ID NO: 30)
31. QEENIKK EE S lEELIKK QEELI IEESI (SEQ ID NO: 31)31. QEENIKK EE S lEELIKK QEELI IEESI (SEQ ID NO: 31)
32. NEESIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 32)32. NEESIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 32)
33. NEEQIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 33)33. NEEQIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 33)
34. SEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 34)34. SEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 34)
35. SEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 35)35. SEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 35)
36. SEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 36)36. SEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 36)
37. QEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 37)37. QEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 37)
38. QEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 38)38. QEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 38)
39. NEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 39)39. NEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 39)
40. NEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 40)40. NEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 40)
41. SEEQIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 41)41. SEEQIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 41)
42. SEENIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 42)42. SEENIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 42)
43. SEESIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 43)43. SEESIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 43)
44. QEESIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 44 )44. QEESIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 44 )
45. QEENIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 45)45. QEENIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 45)
46. NEESIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 46)46. NEESIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 46)
47. NEEQIKK Na風 !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 47)47. NEEQIKK Na Wind !S lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 47)
48. SEEQIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 48)48. SEEQIKK Na Wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 48)
49. SEENIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 49)49. SEENIKK Na wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 49)
50. SEESIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 50)50. SEESIKK Na wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 50)
51. QEESIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 51)51. QEESIKK Na Wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 51)
52. QEENIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 52)52. QEENIKK Na Wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 52)
53. NEESIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 53)53. NEESIKK Na Wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 53)
54. NEEQIKK Na風 Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 54) 55. SEEQIKK QalEEQalS IEELI SEELI IEEQI (SEQ ID NO: 55)；54. NEEQIKK Na Wind Ml lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 54) 55. SEEQIKK Q al EEQ al S IEELI SEELI IEEQI (SEQ ID NO: 55);
56. SEENI QalEEQalS IEELI SEELI IEEQI (SEQ ID NO: 56)； 56. SEENI Q al EEQ al S IEELI SEELI IEEQI (SEQ ID NO: 56);
57. SEESIKK QalEEQalS IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 57)； 57. SEESIKK Q al EEQ al S IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 57);
58. QEESIKK QalEEQalS IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 58)； 58. QEESIKK Q al EEQ al S IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 58);
59. QEENI QalEEQalS IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 59)； 59. QEENI Q al EEQ al S IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 59);
60. NEESI QalEEQalS IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 60)； 60. NEESI Q al EEQ al S IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 60);
61. NEEQI QalEEQalS IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 61)； 61. NEEQI Q al EEQ al S IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 61);
62. SEEQIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 62)； 62. SEEQIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 62);
63. SEENIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 63)； 63. SEENIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 63);
64. SEESIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 64)； 64. SEESIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 64);
65. QEESIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 65)； 65. QEESIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 65);
66. QEENIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 66)； 66. QEENIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 66);
67. NEESIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 67)； 67. NEESIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 67);
68. NEEQIKK QalEEQalD IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 68)； 本发明的另一个方面涉及一种药物组合物， 其含有至少一种上述 的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐， 以及 药学上可接受的载体或辅料。 本发明的还一个方面涉及一种 HIV融合抑制剂， 其含有至少一种 上述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐。 本发明的还一个方面涉及上述的式 I多肽、 其衍生物、 其立体异 构体、 或其无生理毒性的盐在制备 HIV融合抑制剂中的用途。 本发明的还一个方面涉及上述的式 I多肽、 其衍生物、 其立体异 构体、 或其无生理毒性的盐在制备用于治疗或预防 HIV感染相关疾病 尤其是艾滋病的药物中的用途。 本发明的还一个方面涉及一种治疗或预防 HIV感染相关疾病尤其 是艾滋病的方法， 所述的方法包括对给予接受治疗或者预防的对象有 效量的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐。 68. NEEQIKK Q al EEQ al D IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 68); another aspect of the invention relates to a pharmaceutical composition comprising at least one of the above-described polypeptides of formula I, derivatives thereof, A construct, or a salt thereof that is not physiologically toxic, and a pharmaceutically acceptable carrier or adjuvant. A further aspect of the invention relates to an HIV fusion inhibitor comprising at least one of the above-described polypeptides of formula I, derivatives thereof, stereoisomers thereof, or salts thereof which are not physiologically toxic. A further aspect of the invention relates to the use of a polypeptide of the above formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, for the preparation of an HIV fusion inhibitor. A further aspect of the invention relates to the use of a polypeptide of the above formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic for the preparation of a medicament for the treatment or prevention of a disease associated with HIV infection, in particular AIDS. A further aspect of the invention relates to a method of treating or preventing a disease associated with HIV infection, in particular AIDS, comprising administering an effective amount of a polypeptide of formula I, a derivative thereof, or a stereoisomer thereof to a subject to be treated or prevented. Body, or a salt thereof that is not physiologically toxic.
本发明还涉及一种所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐的使用方法，其包括将有效量的所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐施用给有此需要的受 试者。 在本发明中， 术语 "天然" 是指自然存在的， 未经人为修饰的， 如 HIV- 1 gp41本身的氨基酸序列等。  The invention also relates to a method of using the polypeptide of the formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, comprising an effective amount of the polypeptide of the formula I, a derivative thereof The stereoisomer, or a salt thereof that is not physiologically toxic, is administered to a subject in need thereof. In the present invention, the term "natural" refers to a naturally occurring, unmodified, such as the amino acid sequence of HIV-1 gp41 itself.
在本发明中， 术语 "非天然" 是指人为设计的， 与天然不同的， 如对直接衍生的天然序列经改造、 修饰后的序列等。  In the present invention, the term "non-native" means artificially designed, different from nature, such as modified, modified sequences of directly derived natural sequences, and the like.
在本发明中， 术语 "疏水性氨基酸" 是指侧链为疏水性基团的氨 基酸， 包括： Ala, Val, Leu, He, Met, Phe, Trp等。  In the present invention, the term "hydrophobic amino acid" means an amino acid whose side chain is a hydrophobic group, and includes: Ala, Val, Leu, He, Met, Phe, Trp and the like.
在本发明中， 术语 "亲水性氨基酸" 是指侧链含有能与水作用形 成氢键的亲水基团的 ^酸， 包括： Ser， Thr, Cys, Asp, Asn, Glu, Gin, Arg, Lys, His, Tyr等。  In the present invention, the term "hydrophilic amino acid" means an acid having a hydrophilic group capable of forming a hydrogen bond with water, and includes: Ser, Thr, Cys, Asp, Asn, Glu, Gin, Arg , Lys, His, Tyr, etc.
在本发明中， 术语 "酸性氨基酸" 是指侧链含氣基的氨基酸， 包 括： Glu, Asp等。  In the present invention, the term "acidic amino acid" means an amino acid having a gas group in a side chain, and includes: Glu, Asp and the like.
在本发明中， 术语 "碱性 ^酸" 是指侧链含氨基或胍基的 酸， 包括： Lys, Arg等。  In the present invention, the term "basic acid" means an acid having an amino group or a mercapto group in a side chain, and includes: Lys, Arg and the like.
具体实施方式 detailed description
下面将结合实施例对本发明的实施方案进行详细描述， 但是本领 域技术人员将会理解， 下列实施例仅用于说明本发明， 而不应视为限 定本发明的范围。 实施例中未注明具体条件者， 按照常规条件或制造 商建议的条件进行。 所用试剂或仪器未注明生产厂商者， 均为可以通 过市购获得的常规产品。 在本发明中使用的缩写具有下面的含义： The embodiments of the present invention will be described in detail below with reference to the accompanying drawings, however, If no specific conditions are specified in the examples, they are carried out according to the general conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained commercially. The abbreviations used in the present invention have the following meanings:
AIDS (Acquired Immure Deficiency Syndrome) 艾滋病， 获得性免 疫缺陷综合征  AIDS (Acquired Immure Deficiency Syndrome) AIDS, acquired immunodeficiency syndrome
Ala (Alanine, A) 丙氛酸 Ala (Alanine, A) Acetyl
Arg (Arginine, R) 精氛酸Arg (Arginine, R) Sour acid
Asn (Asparagine, N) 天冬醜氛Asn (Asparagine, N) Aspen
Asp (Aspartic acid, D) 天冬氛酸Asp (Aspartic acid, D) Aspartic Acid
DCM (Dichloromethane) 二氯甲坑DCM (Dichloromethane)
DMF (N, N-Dimethyl malonate) 二甲基甲酰胺DMF (N, N-Dimethyl malonate) dimethylformamide
Env (Envelope glycoprotein) 包膜糖蛋白Env (Envelope glycoprotein) envelope glycoprotein
ESI- MS (Electronic spray ion mass spectroscopy) 电喷雾盾谱ESI-MS (Electronic spray ion mass spectroscopy)
Fmoc (F 1 uor eny lme thoxycarbony 1) 甲氧欺基Fmoc (F 1 uor eny lme thoxycarbony 1)
Gly (Glycine, G) 甘氨酸Gly (Glycine, G) glycine
Gin (Glutamine, Q) 谷酰氨Gin (Glutamine, Q) Glutamine
Glu (Glutamic acid, E) 谷氛酸Glu (Glutamic acid, E) glutamic acid
6-HB (six-helix bundle) 六螺旋体 HBTU 2- (1H- 1-羟基苯并三唾 )- 1， 1, 3, 3-四甲基脲六氣磷酸6-HB (six-helix bundle) Hexagonal HBTU 2- (1H- 1-hydroxybenzotris)- 1, 1, 3, 3-tetramethylurea hexa-phosphoric acid
His(Histidine, H) 組氨酸 HOBt (1-Hydroxylbenzotriazole anhydrous) 1 - 基苯并三氮峻His(Histidine, H) histidine HOBt (1-Hydroxylbenzotriazole anhydrous) 1-based benzotriazine
NHR (N-terminal heptad repeat, NHR) N末端重复序列NHR (N-terminal heptad repeat, NHR) N-terminal repeat
CHR (C— terminal heptad repeat, CHR) C末端重复序列CHR (C-terminal heptad repeat, CHR) C-terminal repeat
HIV ( Human Immunodeficiency Virus) ) 人免疫缺陷病毒 HIV- 1 人免疫缺陷病毒 I型HIV (Human Immunodeficiency Virus) Human Immunodeficiency Virus HIV-1 Human Immunodeficiency Virus Type I
HPLC (High performance liquid chromatography) 高效液相色镨HPLC (High performance liquid chromatography)
He (Isoleucine, I) 异亮氨酸He (Isoleucine, I) Isoleucine
Leu (Leucine, L) 亮氨酸 (Nle) 正亮氨酸Leu (Leucine, L) leucine (Nle) norleucine
Lys (Lysine, K) 赖氨酸Lys (Lysine, K) Lysine
Phe (Phenylalanine, F) 苯丙氨酸Phe (Phenylalanine, F) Phenylalanine
Ser (Serine, S) 丝氨酸Ser (Serine, S) Serine
TFA(trif luoroacetic acid) 三氣乙酸TFA(trif luoroacetic acid) trigasacetic acid
Thr (Threonie, T) 苏氨酸Thr (Threonie, T) Threonine
Trp (Tryptophan, W) 色氨酸Trp (Tryptophan, W) Tryptophan
Tyr (Tyrosine, Y) 酪氨酸Tyr (Tyrosine, Y) Tyrosine
Val (Valine, V) 缬氛酸Val (Valine, V) 缬 酸
Nai(Nal) 萘丙氨酸Nai(Nal) naphthylalanine
Qai(Qal) 3-喹啉丙氨酸 腿 P N-甲基吡咯烷酮 DIEA N，N-二异丙基乙胺 实施例所用固相合成载体 Rink酰胺树脂 （ Rink Amide树脂）为 天津南开合成责任有限公司产品； HBTU、 H0BT、 DIEA以及 Fmoc保护 的天然氨基酸为上海吉尔生化公司以及成都诚诺新技术有限责任公 司产品； Fmoc保护的萘丙氨酸及喹啉丙氨酸为本实验室合成产品； N- 甲基吡咯烷酮 （腿 P )为 ACR0S公司产品； 三氟乙酸（TFA )为北京博 迈杰科技有限公司产品； DMF、 DCM为韩国三星公司产品； 色谱纯乙腈 为 Fisher公司产品。 其它试剂如无说明均为国产分析纯产品。 Qai(Qal) 3-quinoline alanine leg P N-methylpyrrolidone The solid phase synthesis carrier Rink Amide resin (Rink Amide resin) used in the DIEA N,N-diisopropylethylamine example is the product of Tianjin Nankai Synthetic Co., Ltd.; the natural amino acid protected by HBTU, H0BT, DIEA and Fmoc is Shanghai Jill Biochemistry. The company and Chengdu Chengnuo New Technology Co., Ltd. products; Fmoc-protected naphthylalanine and quinoline alanine are laboratory synthetic products; N-methylpyrrolidone (leg P) is ACR0S company product; trifluoroacetic acid ( TFA) is a product of Beijing Bomaijie Technology Co., Ltd.; DMF and DCM are products of South Korea's Samsung; chromatographic pure acetonitrile is Fisher's product. Other reagents are domestically produced pure products if they are not described.
实施例 1: SEQ ID NO: 6所示的化合物 6的制备  Example 1: Preparation of Compound 6 as SEQ ID NO: 6
采用标准的 Fmoc 固相多肽合成方法。 所有肽序列均 C端为酰胺 化， N端为乙酰化。 选用 Rink Amide树脂， 氨基酸为 L构型， 肽链由 C端向 N端延长。 活化剂为含有 HBTU和 HOBt的 DMF溶液， 活化碱为 DIEA的腿 P溶液。 脱保护剂为哌啶的 DMF溶液。 裂解剂为三氟乙酸 ( TFA ) ， 粗肽用水溶解后冻干保存。 用中压液相色谱 Flash或高压 液相制备色谱（HPLC )进行分离纯化， 纯肽含量>95%。 基质辅助激光 解析飞行时间质谱（MALDI- T0F- MS )确证多肽序列分子量。  A standard Fmoc solid phase peptide synthesis method was employed. All peptide sequences were amidated at the C-terminus and acetylated at the N-terminus. Rink Amide resin is used, the amino acid is in the L configuration, and the peptide chain is extended from the C terminal to the N terminal. The activator is a DMF solution containing HBTU and HOBt, and the activated base is a leg P solution of DIEA. The deprotecting agent is a DMF solution of piperidine. The cleavage agent is trifluoroacetic acid (TFA), and the crude peptide is dissolved in water and stored by lyophilization. Separation and purification by medium pressure liquid chromatography Flash or high pressure liquid chromatography (HPLC), the pure peptide content is >95%. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-T0F-MS) confirmed the molecular weight of the polypeptide sequence.
使用 CEM全自动微波多肽合成仪进行多肽合成， 合成条件如下： J ^酸： 0. 2M 的 DMF溶液，  The peptide was synthesized using a CEM fully automated microwave peptide synthesizer. The synthesis conditions were as follows: J ^ acid: 0. 2M DMF solution,
活化剂： 0. 45M HBTU/HOBt 的 DMF溶液（其中 HBTU与 HOBt的摩 尔比为 1: 1 ) ，  Activator: 0. 45M HBTU/HOBt DMF solution (where the molar ratio of HBTU to HOBt is 1: 1),
活化碱： 2M DIEA 的腿 P溶液，  Activated base: 2M DIEA leg P solution,
脱保护剂： 20% v/v 哌啶的 DMF溶液，  Deprotecting agent: 20% v/v piperidine in DMF solution,
封闭试剂： 20% v/v 乙酸酐的 DMF溶液。  Blocking reagent: 20% v/v DMF solution of acetic anhydride.
称取 Rink Amide树脂 0. 5g (0. 25mmol)置入 CEM微波多肽合成仪 反应器中， 然后将氨基酸， 活化剂， 活化碱， 脱保护试剂， 封闭试剂 按上述浓度配置好后， 用 CEM微波全自动多肽合成仪进行合成。 完成 后肽树脂用 DMF洗涤 3遍后用无水甲醇收缩， 室温真空干燥， 得肽树 脂 2. 05g。  Rected Rink Amide resin 0. 5g (0. 25mmol) was placed in the CEM microwave peptide synthesizer reactor, and then the amino acid, activator, activated base, deprotection reagent, blocking reagent were arranged at the above concentration, and then CEM microwave was used. Automated peptide synthesizer for synthesis. After completion, the peptide resin was washed with DMF for 3 times, then shrunk with anhydrous methanol, and dried under vacuum at room temperature to obtain a peptide resin of 2.05 g.
裂解液 (体积百分比）：三氟乙酸： 乙二硫醇： 间甲酴： 水 = 82. 5 ： 10 ： 5 ： 2. 5。 Lysate (% by volume): trifluoroacetic acid: ethanedithiol: m-methyl hydrazine: water = 82. 5 : 10 : 5 : 2. 5.
肽树脂的裂解： 称取上述微波合成仪合成好的肽树脂 2. 05g， 放 入 250ml茄形瓶中， 水浴， 电磁搅拌。 按 1克肽树脂加入 10ml的量 配制裂解液。 TFA需预先水浴降温 30min或者预先存放于水箱中使用； 将配制好的裂解液加入到水浴条件下的肽树脂中， 电磁搅拌， 树脂变 橙红色， 水浴条件下反应 30min， 然后， 撤水浴， 室温再继续搅拌反 应 90min， 反应完成。 剧烈搅拌下向反应器中加入冷乙醚 200ml， 析 出白色沉淀， 继续搅拌 30min; 用 G4的砂芯抽虑漏斗滤出析出物， 用 冷乙醚反复洗涤 3遍， 晾干。加入双蒸水 50ml， 乙腈 5ml使固体充分 溶解， 抽虑， 滤液冻干得粗肽 1. 03g。  Pyrolysis of Peptide Resin: Weigh the peptide resin synthesized by the above microwave synthesizer 2. 05g, put it into a 250ml eggplant-shaped bottle, water bath, and electromagnetic stirring. The lysate was prepared by adding 1 gram of the peptide resin to 10 ml. TFA needs to be cooled in a water bath for 30 minutes or pre-stored in a water tank; the prepared lysate is added to the peptide resin under water bath conditions, electromagnetic stirring, the resin turns orange-red, and the reaction is carried out in a water bath for 30 minutes, then, the water bath is removed, room temperature The reaction was further stirred for 90 min and the reaction was completed. 200 ml of cold diethyl ether was added to the reactor under vigorous stirring, and a white precipitate was precipitated, and stirring was continued for 30 min. The precipitate was filtered off with a G4 sand core funnel, washed repeatedly with cold diethyl ether for 3 times, and dried. 50 ml of double distilled water and 5 ml of acetonitrile were added to fully dissolve the solid, and the filtrate was freeze-dried to obtain a crude peptide 1.03 g.
所得粗肽用中压或高压制备型色傳进行纯化。 色傳柱为 C8柱， 洗脱剂为乙腈、 水及少量三氟乙酸。 具体操作步骤： 称取粗肽 1. 00g， 加水 20ml， 乙腈 5ml使固体溶解， 离心 10min (3000转 /分钟）， 取上 清液上样。 色傳柱预先用含有 15% ( v/v ) 乙腈和 0. 1% ( v/v )三氟乙 酸溶液的水溶液 200ml平衡。 上样后在 0-15分钟内继续用含有 15% ( v/v )乙腈和 0. 1% ( v/v )三氟乙酸的水溶液 200ml冲洗， 高效液相 色谱检测洗脱液成分。 根据高效液相色傳检测结果，在 10-30分钟内 将洗脱液中的乙腈含量升高至 65% ( v/v ) (三氟乙酸的含量不变， 保 持 0. 1%ν/ν ) ， 直至所纯化的多肽主峰被洗脱出来。 合并洗脱液， 旋 转蒸发去除有机溶剂， 冻干得纯肽， HPLC检测含量大于 95%。  The resulting crude peptide was purified by medium or high pressure preparative color transfer. The color column is a C8 column, and the eluent is acetonitrile, water and a small amount of trifluoroacetic acid. Specific procedure: Weigh the crude peptide 1. 00g, add water 20ml, acetonitrile 5ml to dissolve the solid, centrifuge for 10min (3000 rev / min), take the supernatant and load. The color column was previously equilibrated with 200 ml of an aqueous solution containing 15% (v/v) acetonitrile and 0.1% (v/v) trifluoroacetic acid solution. After the application, the mixture was washed with 200 ml of an aqueous solution containing 15% (v/v) acetonitrile and 0.1% (v/v) trifluoroacetic acid in 0-15 minutes, and the fraction of the eluate was detected by high performance liquid chromatography. 1%ν/ν。 The acetonitrile content of the eluate was increased to 65% (v / v) in 10-30 minutes (the content of trifluoroacetic acid was unchanged, maintaining 0. 1% ν / ν ) until the main peak of the purified polypeptide is eluted. The eluates were combined, and the organic solvent was removed by rotary evaporation, and the peptide was freeze-dried to obtain a pure peptide content of more than 95% by HPLC.
获得的纯肽经 MALDI- T0F- MS确证其分子量（爾：4477 ) ， 证明所 获得的纯肽为 SEQ ID NO: 6所示的化合物 6。 实施例 2 - 68: SEQ ID NO: 1-5、 7-68 所示的化合物 1- 5， 7-68 的制备  The obtained pure peptide was confirmed by MALDI-TOF-MS to have a molecular weight (el: 4477), and it was confirmed that the obtained pure peptide was the compound 6 shown by SEQ ID NO: 6. Examples 2 - 68: Preparation of the compounds 1- 5, 7-68 of SEQ ID NOS: 1-5, 7-68
SEQ ID NO: 1-5、 7- 68所示的化合物 1-5， 7- 68按实施例 1所述 的方法合成， 其中所述氨基酸均为 L构型， 只是相应的氨基酸残基进 行了替换。 其分子量经 MALDI- T0F- MS确证， 具体数值列于表 1 中。 (见表 1 ) 实施例 69: 抑制 HIV-1生物活性检测 The compounds 1-5, 7-68 shown in SEQ ID NO: 1-5, 7-68 were synthesized as described in Example 1, wherein the amino acids were all in the L configuration, except that the corresponding amino acid residues were replace. The molecular weight was confirmed by MALDI-TOF-MS, and the specific values are shown in Table 1. (See Table 1) Example 69: Inhibition of HIV-1 biological activity assay
化合物抑制 HIV-1介导的细胞 -细胞融合活性评价（ IC5。） ： Compounds inhibit HIV-1 mediated cell-cell fusion activity evaluation (IC 5 .):
1、 实验原理： HIV- 1 慢性感染 H9细胞， 是在 H9细胞内， HIV- 1病毒基因整合到 H9细胞的 DNA链上， 并和 H9细胞一起表达： 将病 毒膜蛋白 GP160表达在 H9细胞的表面。 这时， H9细胞就变成一个巨 大的 HIV- 1病毒。 它的表面膜蛋白 GP160, 在 MT-2细胞的 CD4和趋 同受体 CCR5诱导下， GP160构像发生变化， 变成 GP120和 GP41。 其 诱导 H9细胞和 MT-2细胞发生融合。整个过程可以准确反映 HIV- 1病 毒颗粒与靶细胞融合过程。 受试化合物多肽抑制 HIV- 1感染的 H9细 胞与 MT-2靶细胞的融合， 由此判断受试化合物的抗融合活性。  1. Experimental principle: HIV-1 is chronically infected with H9 cells. In H9 cells, the HIV-1 virus gene is integrated into the DNA strand of H9 cells and expressed together with H9 cells: The viral membrane protein GP160 is expressed in H9 cells. surface. At this time, H9 cells become a huge HIV-1 virus. Its surface membrane protein GP160, induced by CD4 of MT-2 cells and CCR5, changes the conformation of GP160 to GP120 and GP41. It induces fusion of H9 cells and MT-2 cells. The whole process can accurately reflect the fusion process of HIV-1 virus particles with target cells. The test compound polypeptide inhibits fusion of HIV-1 infected H9 cells with MT-2 target cells, thereby judging the anti-fusion activity of the test compound.
2、 染色转移法检测 HIV-1介导的细胞 -细胞融合： HIV-1IIIB感染 的 H9细胞（H9/HIV-1IIIB， 本由实验室自制， 具体方法参见 Jiang, S.， Radigan, L. , and Zhang, L. (2000) Proc. SPIE 3926, 212-219) 被一种荧光试剂 Calcein_AM (Molecular Probes, Inc. , Eugene, OR) 标记，然后， 在 96孔板中每孔加入 50μ1已标记的 H9/HIV-lmB细胞（2 X105个 /ml)， 再加入 50μ1 不同浓度的受试样品（受试样品为实施例2. Detection of HIV-1 mediated cell-cell fusion by dye transfer assay: HIV-1 IIIB- infected H9 cells (H9/HIV-1 IIIB , prepared by the laboratory, see Jiang, S., Radigan, L for specific methods) . and Zhang, L. (2000) Proc. SPIE 3926, 212-219) were labeled with a fluorescent reagent, Calcein_AM (Molecular Probes, Inc., Eugene, OR), and then 50 μl per well was added to a 96-well plate. Labeled H9/HIV-l mB cells (2 X10 5 cells/ml), and then add 50 μl of different concentrations of test samples (test samples are examples)
1-68制备的 SEQ ID NO: 1-68所示的化合物 1-68，从 250 g/ml浓度 两倍梯度稀释）和阳性对照样品 （阳性对照样品为 T-20 和 C34, 从 250 g/ml浓度两倍梯度稀释）， 37°C温育 30min; 然后每孔加入 MT-2 细胞（MT-2为一种常用的效应细胞， 在本实验中用于模拟 HIV感染的 靶细胞， 商购获得， Sigma 公司） 100 u 1 (lxlO6个 /ml)， 37°C温育Compounds 1-68 of SEQ ID NOS: 1-68 prepared by 1-68, diluted from two-fold gradient of 250 g/ml) and positive control samples (positive control samples were T-20 and C34, from 250 g/ MLC concentration was double-diluted), incubated at 37 °C for 30 min; then MT-2 cells were added to each well (MT-2 is a commonly used effector cell, used in this experiment to mimic HIV-infected target cells, commercially available Obtained, Sigma) 100 u 1 (lxlO 6 / ml), 37 ° C incubation
2 h。 2 h.
融合及未融合的 Calcein-AM标记 HIV-1感染的 MT-2细胞用反向 荧光显微镜 (Zeiss, Germany )计数。 应用 CalcuSyn 软件计算 IC5。 值。 Fusion and unfused Calcein-AM labeled HIV-1 infected MT-2 cells were counted using a reverse fluorescence microscope (Zeiss, Germany). Calculate IC 5 using CalcuSyn software. value.
按照上述方法， 测定实施例 1-68制备的 SEQ ID NO: 1-68所示 的化合物 1-68的活性， 其活性测定结果见下面的表 1。 表 1: 多肽抑制 HIV介导的细胞融合活性结果 The activities of the compounds 1-68 shown in SEQ ID NOS: 1-68 prepared in Examples 1-68 were determined according to the methods described above, and the results of the activity assays are shown in Table 1 below. Table 1: Results of polypeptide inhibition of HIV-mediated cell fusion activity
分  Minute
子 抑制活性 IC5。 序 列 量 (μΜ) Subsuppressive activity IC 5 . Sequence quantity (μΜ)
M  M
0. 0026 ± 0. 0003 0. 0026 ± 0. 0003
WEEWDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 1) 4455 WEEWDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 1) 4455
0. 0030 ± 0. 00250. 0030 ± 0. 0025
WEEWRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 2) 4496 WEEWRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 2) 4496
0. 0053 ± 0. 00300. 0053 ± 0. 0030
WEEWSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 3) 4427 WEEWSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 3) 4427
0. 0289 ± 0. 00490. 0289 ± 0. 0049
WEEWIKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 4) 4453 WEEWIKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 4) 4453
0. 0082 ± 0. 00260. 0082 ± 0. 0026
WEEWAKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 5) 4411 WEEWAKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 5) 4411
0. 0657±0. 009260. 0657±0. 00926
N.iEENaiDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 6) 4477 N.iEENaiDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 6) 4477
0. 0086 ± 0. 00060. 0086 ± 0. 0006
N.iEENaiRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 7) 4518 N.iEENaiRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 7) 4518
0. 0071 ± 0. 00030. 0071 ± 0. 0003
N.iEENaiSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 8) 4449 N.iEENaiSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 8) 4449
0. 0345 ± 0· 00370. 0345 ± 0· 0037
N.iEENaiQKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 9) 4490 N.iEENaiQKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 9) 4490
0. 0082 ± 0· 00040. 0082 ± 0· 0004
N.iEENaiNKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 10) 4476 N.iEENaiNKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 10) 4476
0. 0690±0. 00970. 0690±0. 0097
Q.iEEQaiDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 11) 4479 Q.iEEQaiDKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 11) 4479
0. 0570±0. 00460. 0570±0. 0046
Q.iEEQaiRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 12) 4520 Q.iEEQaiRKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 12) 4520
0. 1740±0. 02380. 1740±0. 0238
Q.iEEQaiSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 13) 4451 Q.iEEQaiSKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 13) 4451
0. 0320±0. 00360. 0320±0. 0036
Q.iEEQaiQKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 14) 4492 Q.iEEQaiQKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 14) 4492
0. 1810±0. 06180. 1810±0. 0618
Q.iEEQaiNKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 15) 4478 Q.iEEQaiNKK lEELIKK SEELIKK lEEQIKK QEESIKK (SEQ ID NO: 15) 4478
0. 0082±0. 00070. 0082±0. 0007
SEEQIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 16) 4427 SEEQIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 16) 4427
0. 0142±0. 00250. 0142±0. 0025
SEENIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 17) 4413 0. 0356±0. 0063SEENIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 17) 4413 0. 0356±0. 0063
SEESI WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 18) 4386 SEESI WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 18) 4386
0. 0062±0. 00050. 0062±0. 0005
QEESI WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 19) 4427 QEESI WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 19) 4427
0. 0078±0. 00090. 0078±0. 0009
QEENIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 20) 4454 QEENIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 20) 4454
0. 0042±0. 00250. 0042±0. 0025
NEESIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 21) 4413 NEESIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 21) 4413
0. 0068±0. 00040. 0068±0. 0004
NEEQIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 22) 4454 NEEQIKK WEEWSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 22) 4454
0. 0031±0. 00060. 0031±0. 0006
SEEQI WEEWD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 23) 4455 SEEQI WEEWD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 23) 4455
0. 0058±0. 00040. 0058±0. 0004
SEENI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 24) 4441 SEENI WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 24) 4441
0. 0147±0. 00360. 0147±0. 0036
SEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 25) 4414 SEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 25) 4414
0. 0076±0. 00080. 0076±0. 0008
QEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 26) 4455 QEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 26) 4455
0. 0164±0. 00360. 0164±0. 0036
QEENIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 27) 4482 QEENIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 27) 4482
0. 0379±0. 00370. 0379±0. 0037
NEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 28) 4441 NEESIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 28) 4441
0. 0083±0. 00060. 0083±0. 0006
NEEQIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 29) 4482 NEEQIKK WEEWDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 29) 4482
0. 0089±0. 00070. 0089±0. 0007
QEESIKK WEEWSKK lEELIKK QEELI IEESI (SEQ ID NO: 30) 4427 QEESIKK WEEWSKK lEELIKK QEELI IEESI (SEQ ID NO: 30) 4427
0. 0125±0. 00570. 0125±0. 0057
QEENIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 31) 4454 QEENIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 31) 4454
0. 0168±0. 00870. 0168±0. 0087
NEESIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 32) 4413 NEESIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 32) 4413
0. 0147±0. 00670. 0147±0. 0067
NEEQIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 33) 4454 NEEQIKK WEEWSKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 33) 4454
0. 0093±0. 00070. 0093±0. 0007
SEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 34) 4455 SEEQIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 34) 4455
0. 0143±0. 00280. 0143±0. 0028
SEENIKK WEEWDKK lEELIKK QEELIKK IEESIK (SEQ ID NO: 35) 4441 SEENIKK WEEWDKK lEELIKK QEELIKK IEESIK (SEQ ID NO: 35) 4441
0. 0373±0. 00370. 0373±0. 0037
SEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 36) 4414 0. 0113±0. 0036SEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 36) 4414 0. 0113±0. 0036
QEESIKK WEEWD lEELIKK QEELI IEESI (SEQ ID NO: 37) 4455 QEESIKK WEEWD lEELIKK QEELI IEESI (SEQ ID NO: 37) 4455
0. 0293±0. 00970. 0293±0. 0097
QEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 38) 4482 QEENIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 38) 4482
0. 0478±0. 00820. 0478±0. 0082
NEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 39) 4441 NEESIKK WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 39) 4441
0. 0126±0. 00530. 0126±0. 0053
NEEQI WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 40) 4482 NEEQI WEEWDKK lEELIKK QEELIKK IEESIKK (SEQ ID NO: 40) 4482
0. 0693±0. 00830. 0693±0. 0083
SEEQI NaiEEN.iS lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 41) 4449 SEEQI NaiEEN.iS lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 41) 4449
1. 0147±0. 34671. 0147±0. 3467
SEENI NaiEEN.iSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 42) 4435 SEENI NaiEEN.iSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 42) 4435
3. 0453±0. 56873. 0453±0. 5687
SEESI NaiEEN.iSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 43) 4408 SEESI NaiEEN.iSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 43) 4408
0. 9249±0. 04870. 9249±0. 0487
QEESIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 44) 4449 QEESIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 44) 4449
1. 3557±0. 03751. 3557±0. 0375
QEENIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 45) 4476 QEENIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 45) 4476
0. 0836±0. 05470. 0836±0. 0547
NEESIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 46) 4425 NEESIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 46) 4425
3. 0256±0. 04333. 0256±0. 0433
NEEQIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 47) 4476 NEEQIKK N.iEENaiSKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 47) 4476
0. 0365±0. 00540. 0365±0. 0054
SEEQIKK NaiEEN.iD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 48) 4477 SEEQIKK NaiEEN.iD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 48) 4477
1. 0047±0. 00431. 0047±0. 0043
SEENIKK NaiEEN.iDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 49) 4463 SEENIKK NaiEEN.iDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 49) 4463
2. 0560 ± 0· 03622. 0560 ± 0· 0362
SEESIKK NaiEEN.iDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 50) 4436 SEESIKK NaiEEN.iDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 50) 4436
1. 0048 ± 0. 07351. 0048 ± 0. 0735
QEESIKK N.iEENaiD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 51) 4477 QEESIKK N.iEENaiD lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 51) 4477
1. 0085 ± 0· 08321. 0085 ± 0· 0832
QEENIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 52) 4504 QEENIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 52) 4504
1. 0357 ± 0. 74211. 0357 ± 0. 7421
NEESIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 53) 4463 NEESIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 53) 4463
2. 0347 ± 0· 97352. 0347 ± 0· 9735
NEEQIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 54) 4504 NEEQIKK N.iEENaiDKK lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 54) 4504
0. 0432 ± 0. 00530. 0432 ± 0. 0053
SEEQIKK QaiEEQ.iS lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 55) 4451 56 0. 0848 ± 0. 0072SEEQIKK QaiEEQ.iS lEELIKK SEELIKK lEEQIKK (SEQ ID NO: 55) 4451 56 0. 0848 ± 0. 0072
SEENI QaiEEQ.iS IEELI SEEL IKK IEEQI (SEQ ID NO: 56) 4437 SEENI QaiEEQ.iS IEELI SEEL IKK IEEQI (SEQ ID NO: 56) 4437
57 0. 0344 ± 0. 006557 0. 0344 ± 0. 0065
SEESI QaiEEQ.iSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 57) 4410 SEESI QaiEEQ.iSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 57) 4410
58 0. 0123 ± 0. 009558 0. 0123 ± 0. 0095
QEESI Q.iEEQaiS IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 58) 4451 QEESI Q.iEEQaiS IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 58) 4451
59 0. 3878 ± 0. 006159 0. 3878 ± 0. 0061
QEENI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 59) 4478 QEENI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 59) 4478
60 0. 0154 ± 0. 007760 0. 0154 ± 0. 0077
NEESI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 60) 4437 NEESI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 60) 4437
61 0. 0548 ± 0. 009361 0. 0548 ± 0. 0093
NEEQI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 61) 4478 NEEQI Q.iEEQaiSKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 61) 4478
62 0. 0063 ± 0. 000562 0. 0063 ± 0. 0005
SEEQI QaiEEQ.iD IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 62) 4479 SEEQI QaiEEQ.iD IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 62) 4479
63 0. 0685 ± 0· 053563 0. 0685 ± 0· 0535
SEENIKK QaiEEQ.iDKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 63) 4465 SEENIKK QaiEEQ.iDKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 63) 4465
64 0. 0198 ± 0. 001764 0. 0198 ± 0. 0017
SEESIKK QaiEEQ.iDKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 64) 4438 SEESIKK QaiEEQ.iDKK IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 64) 4438
65 0. 0121 ± 0. 000965 0. 0121 ± 0. 0009
QEESIKK Q.iEEQaiD IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 65) 4479 QEESIKK Q.iEEQaiD IEELIKK SEEL IKK IEEQIKK (SEQ ID NO: 65) 4479
66 0. 0732 ± 0. 037366 0. 0732 ± 0. 0373
QEENIKK Q.iEEQaiDKK IEELIKK SEELI IEEQIKK (SEQ ID NO: 66) 4506 QEENIKK Q.iEEQaiDKK IEELIKK SEELI IEEQIKK (SEQ ID NO: 66) 4506
67 0. 0178 ± 0. 000367 0. 0178 ± 0. 0003
NEESI Q.iEEQaiDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 67) 4465 NEESI Q.iEEQaiDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 67) 4465
68 0. 0068 ± 0. 002768 0. 0068 ± 0. 0027
NEEQIKK Q.iEEQaiDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 68) 4506 NEEQIKK Q.iEEQaiDKK IEELIKK SEELIKK IEEQIKK (SEQ ID NO: 68) 4506
T-20 YTSLIHS LIEESQN QQE NEQ ELLELD WASLWNWF (SEQ ID NO: 69) 0. 0021 ± 0. 0004  T-20 YTSLIHS LIEESQN QQE NEQ ELLELD WASLWNWF (SEQ ID NO: 69) 0. 0021 ± 0. 0004
4491  4491
C34 WMEWDRE INNYTSL IHSLIEE SQNQQE NEQELL (SEQ ID NO: 70) 0. 0063 ± 0. 0012  C34 WMEWDRE INNYTSL IHSLIEE SQNQQE NEQELL (SEQ ID NO: 70) 0. 0063 ± 0. 0012
4289 表 1结果表明，受试化合物（实施例 1-68制备的 SEQ ID NO: 1-68 所示的化合物 1 - 68 )均显示了高的抑制 HIV- 1介导的细胞融合活性。 其中化合物 1-3、 7、 8、 10、 16、 19-24、 26、 29、 30、 39、 62、 68 抑制 HIV- 1介导的细胞融合活性水平（ IC5。 )值与 T- 20以及 C34基本 相当。 4289 The results in Table 1 indicate that the test compounds (Compounds 1-68 of SEQ ID NOS: 1-68 prepared in Examples 1-68) all showed high inhibition of HIV-1 mediated cell fusion activity. Among them, compounds 1-3, 7, 8, 10, 16, 19-24, 26, 29, 30, 39, 62, 68 inhibit HIV-1 mediated cell fusion activity (IC 5 ) values with T-20 And C34 is basically the same.
上述实验证明本发明的式 I所示的多肽能够用于治疗或预防 HIV 感染相关疾病， 尤其是艾滋病。 The above experiments demonstrate that the polypeptide of Formula I of the present invention can be used to treat or prevent HIV Infectious diseases, especially AIDS.
尽管本发明的具体实施方式已经得到详细的描述， 本领域技术人 员将会理解。 根据已经公开的所有教导， 可以对那些细节进行各种修 改和替换， 这些改变均在本发明的保护范围之内。 本发明的全部范围 由所附权利要求及其任何等同物给出。  Although specific embodiments of the invention have been described in detail, those skilled in the art will understand. Various modifications and alterations may be made to those details in accordance with the teachings of the invention, which are within the scope of the invention. The full scope of the invention is indicated by the appended claims and any equivalents thereof.

Claims

1.式 I所示的多肽、 其衍生物、 其立体异构体、 或其无生理毒性 的盐， A polypeptide represented by the formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic,
XalAAXdlXelBB— Xa2AAXd2Xe2BB— XasAAXd3Xe3BB— X AAXd4Xe4BB— XasAAXdsXesBB 式 I XalAAXdlXelBB— X a 2AAXd2Xe2BB— X a sAAXd3Xe3BB — X AAXd4Xe4BB — XasAAXdsXesBB Formula I
其中，  among them,
Xal、 Xa2、 Xa3、 Xa4 、 Xa5分别独立地为 D型或 L型的天然或非天然 的疏水性或亲水性氨基酸； X al, X a2, X a3 , X a4, X a5 each independently a D or L natural or non-natural hydrophobic or hydrophilic amino acid;
A为 D型或 L型的天然或非天然酸性氨基酸；  A is a D or L natural or non-natural acidic amino acid;
Xdl、 Xd2、 Xd3、 Xd4、 Xd5分别独立地为 D型或 L型的天然或非天然的 疏水性氨基酸； X dl , X d2 , X d3 , X d4 , X d5 are each independently a D- or L-type natural or non-natural hydrophobic amino acid;
Xei. Xe2、 X 、 L Xe5分别独立地为 D型或 L型的天然或非天然疏 水性 ^酸； Xei. X e2 , X , LX e5 are each independently a natural or non-natural hydrophobic acid of type D or L;
B为 D型或 L型的天然或非天然碱性氨基酸。  B is a D- or L-type natural or non-natural basic amino acid.
2.根据权利要求 1所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐， 2. A polypeptide of formula I according to claim 1, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic,
其中，  among them,
Xal、 Xa2、 Xa3、 Xa4 、 Xa5分别独立地选自 D型或 L型的如下氨基酸： 色氨酸（W)、萘丙氨酸（Nal)、 3-喹啉丙氨酸（ Qal )、 异亮氨酸（I)、 翔氨酸（V)、 正亮氨酸（Nle)、 亮氨酸（L)、 丙氨酸（A)、 丝氨 酸（ S )、 苏氨酸（ T )、 天冬酰胺（ N )、 和谷氨酰胺（ Q )； X al, X a2, X a3 , X a4, X a5 each independently selected from D or L forms of the following amino acids: tryptophan (W), naphthylalanine (N al), 3- quinoline alanyl Acid (Q al ), isoleucine (I), cineine (V), norleucine (Nle), leucine (L), alanine (A), serine (S), threonine Acid (T), asparagine (N), and glutamine (Q);
A选自 D型或 L型的谷氨酸（E)或天冬氨酸（D) ；  A is selected from D-type or L-form glutamic acid (E) or aspartic acid (D);
Xdl、 Xd2、 Xd3、 Xd4、 Xd5分别独立地选自 D型或 L型的如下氨基酸： 谷酰胺（Q)、 色氨酸（W)、 萘丙氨酸（Nal)、 3-喹啉丙氨酸（Qal)、 异亮氨酸（I) 、 翔氨酸（V) 、 正亮氨酸 、 亮氨酸（L) 、 丙 氨酸（A) 、 天冬酰胺（N) 、 丝氨酸（S) 、 苏氨酸（T) 、 和酪氨酸 (Y) ； Xel. Xe2 , Xe3、 Xe5分别独立地选自 D型或 L型的如下氨基酸： 异亮氨酸（I ) 、 翔氨酸（V ) 、 正亮氨酸 、 亮氨酸（L ) 、 丙 氨酸（A )丝氨酸（S )、 谷酰胺（Q )、 天冬酰胺（N )、 苏氨酸（T )、 和酪氨酸（ Y )； X dl , X d2 , X d3 , X d4 , X d5 are each independently selected from the following amino acids of type D or L: glutamine (Q), tryptophan (W), naphthyl alanine (N al ), 3-quinoline alanine (Q al ), isoleucine (I), cineine (V), norleucine, leucine (L), alanine (A), asparagine ( N), serine (S), threonine (T), and tyrosine (Y); Xel. Xe2 , X e3 and X e5 are each independently selected from the following amino acids of type D or L: isoleucine (I ), cineine (V), norleucine, leucine (L), Alanine (A) serine (S), glutamine (Q), asparagine (N), threonine (T), and tyrosine (Y);
B选自 D型或 L型的赖氨酸（K )或精氨酸（R ) 。  B is selected from lysine (K) or arginine (R) of type D or type L.
3.根据权利要求 1所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐， 其中， 所述式 I多肽选自 SEQ ID NO: 1 - 68所 示序列的多肽中的至少一种。 The polypeptide of the formula I according to claim 1, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, wherein the polypeptide of the formula I is selected from the sequence consisting of SEQ ID NOS: 1 - 68 At least one of the polypeptides.
4.根据权利要求 1所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐， 其中， 所述式 I多肽选自 SEQ ID NO: 1-3、 SEQ ID NO: 7—8、 SEQ ID NO: 10、 SEQ ID NO: 16、 SEQ ID NO: 19—24、 SEQ ID NO: 26、 SEQ ID NO: 29—30、 SEQ ID NO: 39、 SEQ ID NO: 62以及 SEQ ID NO: 68 所示序列的多肽中的至少一种。 The polypeptide of the formula I according to claim 1, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, wherein the polypeptide of the formula I is selected from the group consisting of SEQ ID NO: 1-3, SEQ ID NO: 7-8, SEQ ID NO: 10, SEQ ID NO: 16, SEQ ID NO: 19-24, SEQ ID NO: 26, SEQ ID NO: 29-30, SEQ ID NO: 39, SEQ ID NO: 62 and at least one of the polypeptides of the sequence of SEQ ID NO: 68.
5. 一种药物组合物， 其含有至少一种权利要求 1-4 中任一项所 述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐， 以 及药学上可接受的载体或辅料。 A pharmaceutical composition comprising at least one polypeptide of the formula I according to any one of claims 1 to 4, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, and a pharmaceutically acceptable Acceptable carrier or excipient.
6.一种 HIV融合抑制剂， 其含有至少一种权利要求 1-4中任一项 所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理毒性的盐。 An HIV fusion inhibitor comprising at least one polypeptide of the formula I according to any one of claims 1 to 4, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic.
7.权利要求 1-4中任一项所述的式 I多肽、 其衍生物、 其立体异 构体、 或其无生理毒性的盐在制备 HIV融合抑制剂中的用途。 Use of a polypeptide of formula I according to any one of claims 1 to 4, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, for the preparation of an HIV fusion inhibitor.
8. 权利要求 1-4 中任一项所述的式 I多肽、 其衍生物、 其立体 异构体、或其无生理毒性的盐在制备用于治疗或预防 HIV感染相关疾 病尤其是艾滋病的药物中的用途。 8. The polypeptide of the formula I according to any one of claims 1 to 4, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic in the preparation of a medicament for the treatment or prevention of HIV infection, in particular AIDS Use in medicine.
9. 权利要求 1-4中任一项所述的式 I多肽、 其衍生物、 其立体异 构体、 或其无生理毒性的盐的使用方法， 其包括将有效量的权利要求 1-4中任一项所述的式 I多肽、 其衍生物、 其立体异构体、 或其无生理 毒性的盐施用给有此需要的受试者。 9. A method of using a polypeptide of the formula I according to any one of claims 1 to 4, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic, comprising an effective amount of claims 1-4 The polypeptide of formula I, a derivative thereof, a stereoisomer thereof, or a salt thereof which is not physiologically toxic as described in any one of the above is administered to a subject in need thereof.
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