Identification of Natural Compounds as Inhibitors of Pyruvate Kinase M2 for Cancer Treatment

The reliance of tumor cells on aerobic glycolysis is one of the emerging hallmarks of cancer. Pyruvate kinase M2 (PKM2), an important enzyme of glycolytic pathway, is highly expressed in a number of cancer cells. Tumor cells heavily depend on PKM2 to fulfill their divergent energetic and biosynthetic requirements, suggesting it as novel drug target for cancer therapies. Based on this context, we performed enzymatic-assay-based screening of the in-house phenolic compounds library for the identification of PKM2 inhibitors. This screening identified silibinin, curcumin, resveratrol, and ellagic acid as potential inhibitors of PKM2 with IC₅₀ values of 0.91 µM, 1.12 µM, 3.07 µM, and 4.20 µM respectively. For the determination of Ki constants and the inhibition type of hit compounds, Lineweaver–Burk graphs were plotted. Silibinin and ellagic acid performed the competitive inhibition of PKM2 with Ki constants of 0.61 µM and 5.06 µM, while curcumin and resveratrol were identified as non-competitive inhibitors of PKM2 with Ki constants of 1.20 µM and 7.34 µM. The in silico screening of phenolic compounds against three binding sites of PKM2 provided insight into the binding pattern and functionally important amino residues of PKM2. Further, the evaluation of cytotoxicity via MTT assay demonstrated ellagic acid as potent inhibitor of cancer cell growth (IC₅₀ = 20 µM). These results present ellagic acid, silibinin, curcumin, and resveratrol as inhibitors of PKM2 to interrogate metabolic reprogramming in cancer cells. This study has also provided the foundation for further research to validate the potential of identified bioactive entities for PKM2 targeted-cancer therapies.     

Core-Shell Reactor Partitioning Enzyme and Prodrug by ZIF-8 for NADPH-Sensitive In Situ Prodrug Activation

Enzyme-prodrug therapies have shown unique advantages in efficiency, selectivity, and specificity of in vivo prodrug activation. However, precise spatiotemporal control of both the enzyme and its substrate at the target site, preservation of enzyme activity, and in situ substrate depletion due to low prodrug delivery efficiency continue to be great challenges. Here, we propose a novel core–shell reactor partitioning enzyme and prodrug by ZIF-8, which integrates an enzyme with its substrate and increases the drug loading capacity (DLC) using a prodrug as the building ligand to form a Zn-prodrug shell. Cytochrome P450 (CYP450) is immobilized in ZIF-8, and the antitumor drug dacarbazine (DTIC) is coordinated and deposited in its outer layer with a high DLC of 43.6±0.8 %. With this configuration, a much higher prodrug conversion efficiency of CYP450 (36.5±1.5 %) and lower IC₅₀ value (26.3±2.6 μg/mL) are measured for B16-F10 cells with a higher NADPH concentration than those of L02 cells and HUVECs. With the tumor targeting ability of hyaluronic acid, this core–shell enzyme reactor shows a high tumor suppression rate of 96.6±1.9 % and provides a simple and versatile strategy for enabling in vivo biocatalysis to be more efficient, selective, and safer.     

Synthesis of Cu^Ⅱ/ZIF-8 Metal-organic Framework Catalyst and Its Application in the Aerobic Oxidation of Alcohols

The tliree-dimensional copper-doped zeolitic imidazolate framework ZIF-8(Cu^Ⅱ/ZIF-8) was prepared by a metal ion exchange process, using reaction of three different copper salts, zinc nitrate hexahydrate[Zn(NO)3·6H2O] and 2?methylimidazole(2-MelM) under nitrogen atmosphere at the room temperature. The TEM and PXRD results indicated that the morphology of Cu^Ⅱ/ZIF-8 was rhombic dodecahedron and the structure was intact after copper was doped into the porous ZIF-8. The synthesized Cu(NO3)2/ZIF-8 heterogeneous catalyst showed an excellent activity for tlie aerobic oxidation of primary alcohols employing molecular oxygen as oxidant. Moreover, tlie Cu(NO3)2/ZIF-8 heterogeneous catalyst can cycled 15 times without leaching of copper.     

Enhanced Activity of Enzyme Immobilized on Hydrophobic ZIF-8 Modified by Ni2+ Ions

The development of efficient enzyme immobilization to promote their recyclability and activity is highly desirable. Zeolitic imidazolate framework-8 (ZIF-8) has been proved to be an effective platform for enzyme immobilization due to its easy preparation and biocompatibility. However, the intrinsic hydrophobic characteristic hinders its further development in this filed. Herein, a facile synthesis approach was developed to immobilize pepsin (PEP) on the ZIF-8 carrier by using Ni²⁺ ions as anchor (ZIF-8@PEP-Ni). By contrast, the direct coating of PEP on the surface of ZIF-8 (ZIF-8@PEP) generated significant conformational changes. Electrochemical oxygen evolution reaction (OER) was employed to study the catalytic activity of immobilized PEP. The ZIF-8@PEP-Ni composite attains remarkable OER performance with an ultralow overpotential of only 127 mV at 10 mA cm⁻², which is much lower than the 690 and 919 mV overpotential values of ZIF-8@PEP and PEP, respectively.     

不同方法合成的沸石咪唑酯骨架结构材料(ZIF-8)的表征和催化性能

以二甲基咪唑为有机连接体和以Zn(OH)2或Zn(NO3)2·6H2O为Zn源,在甲醇与氨水的混合溶液、甲醇和DMF 3种不同的合成体系中合成了沸石咪唑酯骨架结构材料ZIF-8(分别记为ZIF-8(NH4OH)、ZIF-8(MeOH)和ZIF-8(DMF),并采用XRD、FTIR、N2吸附、SEM、TPD及Knoevenagel缩合反应等手段对所合成材料进行了表征。结果表明,采用这3种不同的合成方法均可成功制备出ZIF-8,所合成的ZIF-8的形貌基本一致,但其晶粒大小和酸碱性能有较大区别,同ZIF-8(NH4OH)和ZIF-8(DMF)相比,ZIF-8(MeOH)晶粒分布集中、平均粒径较小且具有较大的外比表面积和较多的酸碱位。不同方法合成的ZIF-8在苯甲醛和丙二腈的Knoevenagel缩合反应中的催化性能有很大差异,ZIF-8(MeOH)催化活性明显高于ZIF-8(DMF)和ZIF-8(NH4OH),其较高的催化活性,同其较大的外比表面积和酸碱性能密切相关。     

不同方法合成的沸石咪唑酯骨架结构材料(ZIF-8)的表征和催化性能

以二甲基咪唑为有机连接体和以Zn(OH)₂或Zn(NO₃)₂·6H₂O为Zn源,在甲醇与氨水的混合溶液、甲醇和DMF₃种不同的合成体系中合成了沸石咪唑酯骨架结构材料ZIF-8(分别记为ZIF-8(NH₄OH)、ZIF-8(MeOH)和ZIF-8(DMF),并采用XRD、FTIR、N₂吸附、SEM、TPD及Knoevenagel缩合反应等手段对所合成材料进行了表征。结果表明,采用这3种不同的合成方法均可成功制备出ZIF-8,所合成的ZIF-8的形貌基本一致,但其晶粒大小和酸碱性能有较大区别,同ZIF-8(NH₄OH)和ZIF-8(DMF)相比,ZIF-8(MeOH)晶粒分布集中、平均粒径较小且具有较大的外比表面积和较多的酸碱位。不同方法合成的ZIF-8在苯甲醛和丙二腈的Knoevenagel缩合反应中的催化性能有很大差异,ZIF-8(MeOH)催化活性明显高于ZIF-8(DMF)和ZIF-8(NH₄OH),其较高的催化活性,同其较大的外比表面积和酸碱性能密切相关。     

Self-Assembled pH-Responsive Metal-Organic Frameworks for Enhancing the Encapsulation and Anti-Oxidation and Melanogenesis Inhibition Activities of Glabridin

Metal organic frameworks (MOFs) are formed by self-assembly of metal ions and organic ligands. A special type of MOF called ZIF-8, which is formed by self-assembly of zinc ions and 2-methylimidazole, shows excellent stability in aqueous solutions and disintegrates under acidic conditions. These properties make ZIF-8 a suitable carrier material for pH-stimulated drug delivery systems. Glabridin is an isoflavane compound that is widely present in the roots of licorice. Because of its outstanding skin whitening properties, glabridin is widely used as a whitener in the cosmetics industry. In this study, ZIF-8 was employed to encapsulate glabridin. Glabridin-loaded ZIF-8 was successfully prepared with a drug encapsulation efficiency of 98.67%. The prepared sample showed a fusiform or cruciate flower-like structure, and its size was about 3 μm. ZIF-8 enabled pH-controlled release of glabridin. Moreover, ZIF-8 encapsulation significantly enhanced the intracellular anti-oxidant activity and melanogenesis inhibitory activity of glabridin. This study provides a new approach that shows great potential to improve the biological application of glabridin.     

Selective synthesis of ZIFs from zinc and nickel nitrate solution for photocatalytic H2O2 production

Hydrogen peroxide (H₂O₂) is one of the most promising, green, and effective oxidants that can be used in different applications. In this study, zeolitic imidazolate frameworks (ZIFs), consisting of organic ligands and metal sites, were selectively prepared from zinc or nickel nitrate solutions for use in photocatalytic H₂O₂ production. High concentrations of zinc nitrate solution provided more metal sites to coordinate with 2-methylimidazole, producing ZIF-8 with larger particle size, whereas low zinc nitrate concentrations resulted in more interconnected N–H⋯N hydrogen bonds, forming 2D-layered ZIF-L, with smaller particle size. Various concentrations of zinc and nickel nitrate solutions produced ZIFs that exhibited ZIF-8 or ZIF-L topology, with bandgap energies of 5.45 and 4.85 eV, respectively. These samples could serve as promising photocatalyst for the successful production of H₂O₂ under Xenon lamp irradiation.     

ZIF-8 Nanoparticles Evoke Pyroptosis for High-Efficiency Cancer Immunotherapy

Although zeolitic imidazolate framework-8 (ZIF-8) has been applied in various tumor therapies, the intrinsic immunogenicity remains unclear. Here, we initiatively discover that ZIF-8 nanoparticles (NPs) can intrinsically induce pyroptosis by a caspase-1/gasdermin D (GSDMD)-dependent pathway. The pyroptotic cell death is accompanied by necrosis and immunogenic cell death (ICD) simultaneously for efficient in situ immunity initiation. Meanwhile, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), a mitochondrial depolarizing agent, is successfully loaded into ZIF-8 NPs and found to further enhance the pyroptosis process. Collectively, the obtained Pluronic F127-modified CCCP-incorporated ZIF-8 NPs (^F127ZIF-8_CCCP NPs) activate antitumor immunity and reprogram immunosuppressive tumor microenvironment (TME), realizing high-efficiency tumor growth inhibition. This work will facilitate biomedicine applications of ZIF-8 and provide good inspiration for pyroptosis-induced cancer therapy.     

Experimental and molecular simulation studies of CO2 adsorption on zeolitic imidazolate frameworks: ZIF-8 and amine-modified ZIF-8

ZIF-8 has been rapidly developed as a potential candidate for CO₂ capture due to its low density, high surface area, and robust structure. Considering the electron-donating effect of amino functional groups, amino-modification is expected to be an efficient way to improve CO₂ adsorption of ZIF-8. In this work, grand canonical Monte Carlo (GCMC) simulation was performed to study the CO₂ adsorption isotherm based on ZIF-8, ZIF-8-NH₂, and ZIF-8-(NH₂)₂. ZIF-8 was synthesized and CO₂ adsorption isotherms based on ZIF-8 was measured. The experimental surface area, pore volume, and CO₂ adsorption isotherm were used to validate the force field. Adsorptive capacity of ZIF-8-NH₂, and ZIF-8-(NH₂)₂ were first estimated. The GCMC simulation results indicated that the order of increasing CO₂ capacity of the ZIF-8 in the lower pressure regime is: ZIF-8 < ZIF-8-NH₂ < ZIF-8-(NH₂)₂, and in the high pressure is: ZIF-8 < ZIF-8-(NH₂)₂ < ZIF-8-NH₂. New adsorption sites can be generated with the existence of-NH₂ groups. In addition, for non-modified and amino-modified ZIF-8, it was the first time to use density functional theory (DFT) calculations to investigate their CO₂ adsorption sites and CO₂ binding energies. The present work indicates that appropriate amine-functionalized can directly enhanced CO₂ capacity of ZIF-8.     

ZIF-8原位负载Pd纳米粒子及其在Suzuki-Miyaura反应中的应用(英文)

将PdCl2与ZIF-8的反应原料ZnO和2-甲基咪唑按照一定的比例,采用机械化学法原位将Pd2+负载在ZIF-8上(Pd2+/ZIF-8)。然后用NaBH4将Pd2+/ZIF-8进行还原,得到均匀分散的Pd纳米颗粒(Pd/ZIF-8)。通过XRD、N2吸附、透射电镜、ICP-AES、XPS等对Pd/ZIF-8的结构、形貌、价态等进行了表征。结果表明用机械化学法原位制备的Pd/ZIF-8具有分散均匀、容易大量制备的优点。该催化剂不仅能高效催化Suzuki-Miyaura交叉偶联反应,并且能够多次循环利用。     

Fine-Tuning Electrolyte Concentration and Metal–Organic Framework Surface toward Actuating Fast Zn2+ Dehydration for Aqueous Zn-Ion Batteries

Functional porous coating on zinc electrode is emerging as a powerful ionic sieve to suppress dendrite growth and side reactions, thereby improving highly reversible aqueous zinc ion batteries. However, the ultrafast charge rate is limited by the substantial cation transmission strongly associated with dehydration efficiency. Here, we unveil the entire dynamic process of solvated Zn²⁺ ions’ continuous dehydration from electrolyte across the MOF-electrolyte interface into channels with the aid of molecular simulations, taking zeolitic imidazolate framework ZIF-7 as proof-of-concept. The moderate concentration of 2 M ZnSO₄ electrolyte being advantageous over other concentrations possesses the homogeneous water-mediated ion pairing distribution, resulting in the lowest dehydration energy, which elucidates the molecular mechanism underlying such concentration adopted by numerous experimental studies. Furthermore, we show that modifying linkers on the ZIF-7 surface with hydrophilic groups such as −OH or −NH₂ can weaken the solvation shell of Zn²⁺ ions to lower the dehydration free energy by approximately 1 eV, and may improve the electrical conductivity of MOF. These results shed light on the ions delivery mechanism and pave way to achieve long-term stable zinc anodes at high capacities through atomic-scale modification of functional porous materials.     

ZIF-8基复合材料高效、稳定电催化还原CO2

电催化CO₂还原反应(eCO₂RR)受到催化剂本征活性以及传质的限制,导致材料的催化活性低、反应起始电位高等问题。我们以类沸石锌盐咪唑骨架(ZIF-8)材料为研究对象,探究了不同粒径ZIF-8材料的eCO₂RR性能。优选粒径为50 nm的ZIF-8材料,进一步引入碳纳米管(CNT)作为其导电基底材料,通过原位生长,构建了复合材料ZIF-8-50@CNT的多级孔结构和疏水界面。eCO₂RR实验结果表明,CNT的引入提高了催化剂的导电性,优化后的复合材料有效地降低了反应的起始电位。在-1.1 V(相对可逆氢电极(RHE))电位下,CO部分电流密度为15.6 mA·cm^-2,ZIF-8-50@CNT催化剂的比表面活性提升了3.5倍(相比ZIF-8-50),塔菲尔斜率降低到136 mV·dec^-1。并且产物CO的选择性和稳定性得到了提高,在宽电势窗口-0.9～-1.2 V(vs RHE)内,CO的法拉第效率(FE)保持在80%以上。在10 h稳定性测试中,催化剂活...     

ZIF-8基复合材料高效、稳定电催化还原CO2

电催化CO₂还原反应(eCO₂RR)受到催化剂本征活性以及传质的限制，导致材料的催化活性低、反应起始电位高等问题。我们以类沸石锌盐咪唑骨架(ZIF-8)材料为研究对象，探究了不同粒径ZIF-8材料的eCO₂RR性能。优选粒径为50 nm的ZIF-8材料，进一步引入碳纳米管(CNT)作为其导电基底材料，通过原位生长，构建了复合材料ZIF-8-50@CNT的多级孔结构和疏水界面。eCO₂RR实验结果表明，CNT的引入提高了催化剂的导电性，优化后的复合材料有效地降低了反应的起始电位。在-1.1 V (相对可逆氢电极(RHE))电位下，CO部分电流密度为15.6 mA·cm^-2，ZIF-8-50@CNT催化剂的比表面活性提升了3.5倍(相比ZIF-8-50)，塔菲尔斜率降低到136 mV·dec^-1。并且产物CO的选择性和稳定性得到了提高，在宽电势窗口-0.9~-1.2 V (vs RHE)内，CO的法拉第效率(FE)保持在80%以上。在10 h稳定性测试中，催化剂活性保持稳定，整体增强了复合材料eCO₂RR的性能。     

Metal–organic frameworks as efficient materials for drug delivery: Synthesis,characterization, antioxidant,anticancer, antibacterial and molecular docking investigation

Metal–organic framework (MOF) nano particles are a class of promising porous nano materials for biomedical applications. Owing to its high loading potential and pH-sensitive degradation, most promising of the MOFs is the zeolitic imidazolate crystal framework (ZIF-8), a progressive useful material for small molecule distribution. Doxorubicin (DOX), designated as a classical drug, was jobwise entrapped in ZIF-8 nano particles. ZIF-8 nano particles, as a novel carrier, were used to monitor the release of the anticancer drug DOX and prevent it from dissipating before reaching its goal. ZIF-8 nano particles with encapsulated DOX (DOX@ZIF-8) can be synthesized in a single pot by incorporation of DOX into the reaction mixture. MOFs and the designed drug delivery (DOX@ZIF-8) system were characterized by Fourier transfer infrared, scanning electron microscopy, N₂ sorption isotherm and X-ray diffraction. The impact of MOFs and the engineered drug delivery system on the viability of human breast and liver cancer cell lines was evaluated. The loaded drug was released at pH 5 faster than at pH 7.4. The nano particles of ZIF-8 showed low cytotoxicity, while DOX@ZIF-8 showed high cytotoxicity to HepG-2 and MCF-7 cells compared with free DOX at the equivalent concentration of DOX of >12.5 μg/ml. These findings indicate that DOX@ZIF-8 nano particles are a promising method for the delivery of cancer cells to drugs. Furthermore, ZIF-8, DOX and encapsulated DOX@ZIF-8 compounds were screened for their potential antibacterial activities against pathogenic bacteria compared with standard antibiotics by the agar well diffusion technique. The results demonstrate that the DOX@ZIF-8 exhibits a strong inhibition zone against Gram-negative strains (Escherichia coli) in comparison with the reference drug gentamycin. The docking active site interactions were evaluated to predict the binding between DOX with the receptor of breast cancer 3hb5-oxidoreductase and liver cancer 2h80-lipid binding protein for anticancer activity.     

Selective cyclodimerization of epichlorohydrin to dioxane derivatives over MOFs

Glycerol can be converted to valuable products such as epichlorohydrin which is an important intermediate applied in various industries. For example, dioxane derivatives, which are important pharmaceuticals, can be obtained from epichlorohydrin. In the present study, ZIF-8, ZIF-67, MIL-100, and UiO-66 were applied for the direct cyclodimerization of epichlorohydrin. These MOFs were selected because they were already applied as active catalysts in ring opening of epoxides. Among them, ZIF-8 showed the highest activity and selectivity in the absence of any solvent or co-catalyst. Using ZIF-8 as a catalyst, the cyclodimer product (1,4-dioxane 2,5-bis-chloromethyl) was obtained in a yield of about 70% which was significantly superior to previous homo or heterogeneous catalysts for this reaction. Due to ZIF-8 structure and the proposed mechanism, the cyclodimerization reaction catalyzed either by the defects in the structure and/or on the surface. Furthermore, acidic-basic characteristics were also in play. The NH₃ and CO₂ temperature-programed desorption technique were utilized to identify the active sites and thereby reaction mechanism. Moreover, because of similar properties of ZIF-8 to zeolites, the activity of commercial ZSM-5 for the same reaction was also investigated in this work.     

Influence of Substitutional Defects in ZIF-8 Membranes on Reverse Osmosis Desalination: A Molecular Dynamics Study

In this study, molecular dynamics simulation is used to investigate the effects of water-based substitutional defects in zeolitic imidazolate frameworks (ZIF)-8 membranes on their reverse osmosis (RO) desalination performance. ZIF-8 unit cells containing up to three defect sites are used to construct the membranes. These substitutional defects can either be Zn defects or linker defects. The RO desalination performance of the membranes is assessed in terms of the water flux and ion rejection rate. The effects of defects on the interactions between the ZIF-8 membranes and NaCl are investigated and explained with respect to the radial distribution function (RDF) and ion density distribution. The results show that ion adsorption on the membranes occurs at either the nitrogen atoms or the defect sites. Complete NaCl rejection can be achieved by introducing defects to change the size of the pores. It has also been discovered that the presence of linker defects increases membrane hydrophilicity. Overall, molecular dynamics simulations have been used in this study to show that water-based substitutional defects in a ZIF-8 structure reduce the water flux and influence its hydrophilicity and ion adsorption performance, which is useful in predicting the type and number of defect sites per unit cell required for RO applications. Of the seven ZIF-8 structures tested, pristine ZIF-8 exhibits the best RO desalination performance.     

A Combined Experimental and Computational Study on the Adsorption Sites of Zinc-Based MOFs for Efficient Ammonia Capture

Ammonia (NH₃) is a common pollutant mostly derived from pig manure composting under humid conditions, and it is absolutely necessary to develop materials for ammonia removal with high stability and efficiency. To this end, metal–organic frameworks (MOFs) have received special attention because of their high selectivity of harmful gases in the air, resulting from their large surface area and high density of active sites, which can be tailored by appropriate modifications. Herein, two synthetic metal–organic frameworks (MOFs), 2-methylimidazole zinc salt (ZIF-8) and zinc-trimesic acid (ZnBTC), were selected for ammonia removal under humid conditions during composting. The two MOFs, with different organic linkers, exhibit fairly distinctive ammonia absorption behaviors under the same conditions. For the ZnBTC framework, the ammonia intake is 11.37 mmol/g at 298 K, nine times higher than that of the ZIF-8 framework (1.26 mmol/g). In combination with theoretical calculations, powder XRD patterns, FTIR, and BET surface area tests were conducted to reveal the absorption mechanisms of ammonia for the two materials. The adsorption of ammonia on the ZnBTC framework can be attributed to both physical and chemical adsorption. A strong coordination interaction exists between the nitrogen atom from the ammonia molecule and the zinc atom in the ZnBTC framework. In contrast, the absorption of ammonia in the ZIF-8 framework is mainly physical. The weak interaction between the ammonia molecule and the ZIF-8 framework mainly results from the inherent severely steric hindrance, which is related to the coordination mode of the imidazole ligands and the zinc atom of this framework. Therefore, this study provides a method for designing promising MOFs with appropriate organic linkers for the selective capture of ammonia during manure composting.     

Catalytically Active Hollow Fiber Membranes with Enzyme-Embedded Metal–Organic Framework Coating

Metal–organic frameworks (MOFs) are suitable enzyme immobilization matrices. Reported here is the in situ biomineralization of glucose oxidase (GOD) into MOF crystals (ZIF-8) by interfacial crystallization. This method is effective for the selective coating of porous polyethersulfone microfiltration hollow fibers on the shell side in a straightforward one-step process. MOF layers with a thickness of 8 μm were synthesized, and fluorescence microscopy and a colorimetric protein assay revealed the successful inclusion of GOD into the ZIF-8 layer with an enzyme concentration of 29±3 μg cm⁻². Enzymatic activity tests revealed that 50 % of the enzyme activity is preserved. Continuous enzymatic reactions, by the permeation of β-d -glucose through the GOD@ZIF-8 membranes, showed a 50 % increased activity compared to batch experiments, emphasizing the importance of the convective transport of educts and products to and from the enzymatic active centers.     

A new class of HIV-1 inhibitors and the target identification via proteomic profiling

Anti-HIV screening with the MT-4/MTT assay on a focused library of structurally diverse natural products has led to the discovery of a group of steroids with potent activities, which include four new ergostane-type steroids, named amotsterols A-D (1-4), together with two known analogs. Among them, the most potent amotsterol D (4) exhibited anti-HIV activity against wildtype and some clinically relevant multidrug resistant HIV-1 strains. Subsequent studies on its target identification through a proteomic approach found that compound 4 might target PKM2, a rate limiting enzyme of glycolysis, in host cells to restrict HIV replication. The docking model of compound 4 to PKM2 showed that the two hydroxyl groups of 4 form hydrogen bonds with the two parallel Y390 in each subunit of PKM2 separately, and the ring C of 4 is sandwiched between the two parallel aromatic rings of F26. The identified hit compound may have the potential to be further developed as a novel anti-HIV agent. These results demonstrated that an integrated approach, which combines new chemical structures and phenotypic screening with a proteomic approach, could not only identify novel HIV-1 inhibitors, but also elucidate the unknown targets of compound interactions in antiviral drug discovery.