Development of Antiviral Therapy of HIV-1 Infection

HIV-1感染抗病毒治疗的进展

基本信息

批准号：
10262697
负责人：
Hiroaki Mitsuya
金额：
$ 75.77万
依托单位：
DIVISION OF CLINICAL SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10262697
关键词：
3-Dimensional Acquired Immunodeficiency Syndrome Active Sites Affect Affinity Amino Acid Substitution Amino Acids Antiviral Agents Antiviral Therapy Benzene Benzoxazoles Binding Biological Availability CCR Canis familiaris Cell Membrane Permeability Cells Chemicals Cleaved cell Clinical Collaborations Complex Data Dental crowns Development Distal Drug Design Drug Kinetics Drug resistance Drug usage Elements Enzymes FDA approved Fluorine Future Generations Genetic Glycine HIV HIV-1 HIV-1 drug resistance HIV-1 protease Halogens Hydrogen Bonding Hydrophobicity Immune system Infection Ligands Membrane Metabolic Modification Molecular Conformation Multi-Drug Resistance Oral Paper Penetration Peptide Hydrolases Pharmaceutical Preparations Pharmacologic Substance Positioning Attribute Property Protease Inhibitor Proteins RNA-Directed DNA Polymerase Rattus Reporting Resistance Roentgen Rays Role Scanning Series Site Structure Sulfur Surface Surgical Flaps Therapeutic Toxic effect United States National Institutes of Health Universities Variant Vertebral column Viral Viral Physiology Virion Virus antiretroviral therapy base benzothiazole blood-brain barrier permeabilization clinical development design dimer drug development drug resistant virus flexibility inhibitor/antagonist nanomolar novel novel therapeutics pandemic disease peptidomimetics pol Gene Products professor prototype restoration scaffold

项目摘要

In the current project, we demonstrated that fine-tuning the DRV structure, e.g., by replacement of a cyclopropyl with a saturated isopropyl moiety and/or the introduction of a fluorine atom(s), alters the potency against HIV-1. In particular, the present membrane penetration data confirm and strengthen our previous findings that the addition of two fluorine atoms greatly boosts the inhibitory properties of the PIs examined. We have also examined to what extent the position of such fluorines affect their anti-HIV-1 activity. In addition to the effects described, dual fluorine substitutions may alter metabolic stability and cellular toxicity, but these parameters were not examined in detail in the present study. The percentage of fluorinated therapeutics in the pharmaceutical market has markedly increased over the decades and in 2019, four out of ten approved new drugs contains at least one fluorine atom. Among the nine FDA-approved PIs, however, only TPV contains three fluorines. However, fluorine scan is increasingly being used for drug development. To this end, our data may further contribute to the use of fluorine substitutions in the future design of novel PIs. It was also noted in the current study that the inhibitors containing benzothiazole moiety at the P2' position exerted greater anti-HIV-1 activity than those with benzoxazole moiety. This greater potency owes to the capacity of sulfur atoms. Sulfur is indeed, one of the most prominent atoms in the chemical composition of FDA approved drugs.30 Recent analyses of sulfur bonding interactions based on PDB structures has shown sulfur-based interactions most often take place with glycine backbone due to lack of steric hindrance, which is apparently what we have seen with sulfur-Gly48 interaction. Furthermore, the cyclopropyl-containing inhibitors exerted quite robust activity against most of the drug-resistant HIV-1 variant examined here. Replacement of cyclopropyl with isopropyl at the distal part of the inhibitor's P2' moiety results in a reduction in the anti-viral activity against wild-type HIV-1. In addition to well-recognized positive properties of cyclopropyl substitutions in drug design, such as increased metabolic stability and cell and blood-brain-barrier permeability, our findings make an example of optimization approaches particularly against multi drug resistant targets. Comparison between cyclo- and iso-propyl groups in regard to cell membrane permeability remains to be elucidated; however, we observed a greater membrane permeability of compounds with an isopropyl group in the current study. The majority of the FDA-approved PIs including DRV have peptidomimetic features and our extended and continuous efforts here resulted in a new set of inhibitors comprising more complex chemical arrangements derived from modifications on three moieties. Of note, the P2-crn-THF moiety occupies a larger three-dimensional space compared to DRV's P2-bis-THF moiety. The P2'-cyclopropyl moiety can rotate and therefore adapt to a better binding mode in the hydrophobic cavity of PR. Both crn-THF and benzothiazole moieties also snugly fill up the hydrophobic tunnel formed between HIV-1 protease dimer, resulting in extensive vdW interactions with HIV-1 protease compared to the case of DRV. Although the X-ray-based structural differences between PRWT and PRDRVRs appear to be rather small and subtle, such differences are apparently sufficient to reduce the affinity of PIs to PR. By comparing the structures of PRWT with PRDRVRs, we found various plausibly significant conformational changes which affect the binding pocket. Consequently, inhibitors have to adapt to the increased flexibilities of target proteins. Particularly, bis-meta fluorine and benzothiazole 2-amino cyclopropyl groups are indispensable components to sustain potent antiviral activity particulary against multi-drug resistant viruses. As a result of optimizing the P1 and P2' positions, the prototypic DRV has been converted to exceedingly potent PIs such as GRL-063 and GRL-016 as well as GRL-142, GRL-001, and GRL-003. In our previous paper reporting the generation of HIVDRVRP30, we determined all the gag-encoding regions of HIVDRVRP30 and other variants. However, a variety of amino acid substitutions were present at the outset in the mixture of 8 highly drug-resistant clinical isolates and more amino acid substitutions emerged under selection; thereby we had to choose not to pursue to determine the role of each amino acid substitution in the emergence of resistance to DRV. For the same reason, we did not pursue to determine the role of each amino acid substitution in the gag region in the present study. Pharmacokinetics studies of TMC-126, a prototype of TMC-114 (also known as DRV), with rats and dogs showed that oral bioavailability of TMC-126 was too low to warrant further clinical development. Further optimization of the series resulted in the selection of TMC-114 due to its superior pharmacokinetics and antiviral profile in comparison with all the other compounds of the series such as TMC-126. The aforementioned features of GRL-001 strongly suggest that the compound is a promising candidate as a novel anti-HIV therapeutic and that GRL-001 should be investigated further for potential clinical development. In particular, GRL-001 appears to tolerate amino acid substitutions in the PR bindings pocket, making them promising PIs for future clinical development.

在当前的项目中，我们证明了微调 DRV 结构，例如通过用饱和异丙基部分替换环丙基和/或引入氟原子，可以改变抗 HIV-1 的效力。特别是，目前的膜渗透数据证实并加强了我们之前的发现，即添加两个氟原子大大增强了所检测的 PI 的抑制特性。我们还研究了这些氟的位置在多大程度上影响其抗 HIV-1 活性。除了所描述的影响之外，双氟取代可能会改变代谢稳定性和细胞毒性，但本研究中没有详细检查这些参数。几十年来，氟化药物在药品市场中的比例显着增加，2019 年，十分之四的批准新药含有至少一个氟原子。然而，在 FDA 批准的九种 PI 中，只有 TPV 含有三种氟。然而，氟扫描越来越多地用于药物开发。为此，我们的数据可能进一步有助于在未来新型 PI 设计中使用氟替代品。本研究还指出，P2'位置含有苯并噻唑部分的抑制剂比含有苯并恶唑部分的抑制剂具有更强的抗HIV-1活性。这种更大的效力归功于硫原子的能力。硫确实是 FDA 批准的药物化学成分中最重要的原子之一。 30 最近基于 PDB 结构对硫键相互作用的分析表明，由于缺乏空间位阻，硫基相互作用最常与甘氨酸主链发生，这显然是我们在硫-Gly48 相互作用中所看到的。此外，含环丙基的抑制剂对本文检查的大多数耐药 HIV-1 变体发挥了相当强的活性。在抑制剂 P2' 部分的远端用异丙基取代环丙基会导致针对野生型 HIV-1 的抗病毒活性降低。除了环丙基取代在药物设计中众所周知的积极特性（例如增加代谢稳定性以及细胞和血脑屏障渗透性）外，我们的研究结果还为特别针对多重耐药靶点的优化方法提供了一个例子。环丙基和异丙基在细胞膜渗透性方面的比较仍有待阐明；然而，我们在当前的研究中观察到具有异丙基的化合物具有更大的膜渗透性。大多数 FDA 批准的 PI（包括 DRV）都具有拟肽特征，我们在此进行的长期持续努力产生了一组新的抑制剂，其中包含源自三个部分修饰的更复杂的化学排列。值得注意的是，与 DRV 的 P2-bis-THF 部分相比，P2-crn-THF 部分占据更大的三维空间。 P2'-环丙基部分可以旋转，因此适应 PR 疏水腔中更好的结合模式。 crn-THF 和苯并噻唑部分也紧密地填充了 HIV-1 蛋白酶二聚体之间形成的疏水通道，与 DRV 的情况相比，导致 vdW 与 HIV-1 蛋白酶发生广泛的相互作用。尽管 PRWT 和 PRDRVR 之间基于 X 射线的结构差异似乎相当小且微妙，但这种差异显然足以降低 PI 与 PR 的亲和力。通过比较 PRWT 与 PRDRVR 的结构，我们发现了影响结合袋的各种看似显着的构象变化。因此，抑制剂必须适应靶蛋白增加的灵活性。特别地，双间氟和苯并噻唑2-氨基环丙基是维持有效的抗病毒活性，特别是针对多重耐药病毒的不可缺少的成分。通过优化 P1 和 P2' 位置，原型 DRV 已转化为极其有效的 PI，例如 GRL-063 和 GRL-016 以及 GRL-142、GRL-001 和 GRL-003。在我们之前报告 HIVDRVRP30 生成的论文中，我们确定了 HIVDRVRP30 和其他变体的所有 gag 编码区域。然而，8个高度耐药的临床分离株的混合物中一开始就存在多种氨基酸取代，并且在选择中出现了更多的氨基酸取代；因此，我们不得不选择不去确定每个氨基酸取代在 DRV 抗性出现中的作用。出于同样的原因，我们在本研究中没有试图确定 gag 区域中每个氨基酸取代的作用。 TMC-126（TMC-114（也称为 DRV）的原型）在大鼠和狗身上的药代动力学研究表明，TMC-126 的口服生物利用度太低，无法保证进一步的临床开发。该系列的进一步优化导致选择 TMC-114，因为与该系列的所有其他化合物（如 TMC-126）相比，TMC-114 具有优异的药代动力学和抗病毒特性。 GRL-001 的上述特征强烈表明该化合物是一种有前途的候选新型抗 HIV 治疗药物，并且应该进一步研究 GRL-001 以进行潜在的临床开发。特别是，GRL-001 似乎能够耐受 PR 结合袋中的氨基酸取代，这使得它们成为未来临床开发的有前景的 PI。