Synthesis and in vivo Evaluation of Fluorine‐18 and Iodine‐123 Pyrazolo[4,3‐e]‐1,2,4‐triazolo[1,5‐c]pyrimidine Derivatives as PET and SPECT Radiotracers for Mapping A2A Receptors

Imaging agents that target adenosine type 2A (A_2A) receptors play an important role in evaluating new pharmaceuticals targeting these receptors, such as those currently being developed for the treatment of movement disorders like Parkinson′s disease. They are also useful for monitoring progression and treatment efficacy by providing a noninvasive tool to map changes in A_2A receptor density and function in neurodegenerative diseases. We previously described the successful evaluation of two A_2A‐specific radiotracers in both nonhuman primates and in subsequent human clinical trials: [¹²³I]MNI‐420 and [¹⁸F]MNI‐444. Herein we describe the development of both of these radiotracers by selection from a series of A_2A ligands, based on the pyrazolo[4,3‐e]‐1,2,4‐triazolo[1,5‐c]pyrimidine core of preladenant. Each of this series of 16 ligands was found to bind to recombinant human A_2A receptor in the low nanomolar range, and of these 16, six were radiolabeled with either fluorine‐18 or iodine‐123 and evaluated in nonhuman primates. These initial in vivo results resulted in the identification of 7‐(2‐(4‐(4‐(2‐[¹⁸F]fluoroethoxy)phenyl)piperazin‐1‐yl)ethyl)‐2‐(furan‐2‐yl)‐7H‐pyrazolo[4,3‐e][1,2,4]triazolo[1,5‐c]pyrimidin‐5‐amine ([¹⁸F]MNI‐444) and 7‐(2‐(4‐(2‐fluoro‐4‐[¹²³I]iodophenyl)piperazin‐1‐yl)ethyl)‐2‐(furan‐2‐yl)‐7H‐imidazo[1,2‐c]pyrazolo[4,3‐e]pyrimidin‐5‐amine ([¹²³I]MNI‐420) as PET and SPECT radiopharmaceuticals for mapping A_2A receptors in brain.     

Development of 18F-Labeled Radiotracers for PET Imaging of the Adenosine A2A Receptor: Synthesis,Radiolabeling and Preliminary Biological Evaluation

The adenosine A_2A receptor (A_2AR) represents a potential therapeutic target for neurodegenerative diseases. Aiming at the development of a positron emission tomography (PET) radiotracer to monitor changes of receptor density and/or occupancy during the A_2AR-tailored therapy, we designed a library of fluorinated analogs based on a recently published lead compound (PPY). Among those, the highly affine 4-fluorobenzyl derivate (PPY1; K_i(hA_2AR) = 5.3 nM) and the 2-fluorobenzyl derivate (PPY2; K_i(hA_2AR) = 2.1 nM) were chosen for ¹⁸F-labeling via an alcohol-enhanced copper-mediated procedure starting from the corresponding boronic acid pinacol ester precursors. Investigations of the metabolic stability of [¹⁸F]PPY1 and [18F]PPY2 in CD-1 mice by radio-HPLC analysis revealed parent fractions of more than 76% of total activity in the brain. Specific binding of [¹⁸F]PPY2 on mice brain slices was demonstrated by in vitro autoradiography. In vivo PET/magnetic resonance imaging (MRI) studies in CD-1 mice revealed a reasonable high initial brain uptake for both radiotracers, followed by a fast clearance.     

Binding Kinetics of ZM241385 Derivatives at the Human Adenosine A2A Receptor

Classical drug design and development rely mostly on affinity‐ or potency‐driven structure–activity relationships (SAR). Thus far, a given compound’s binding kinetics have been largely ignored, the importance of which is now being increasingly recognized. In the present study, we performed an extensive structure–kinetics relationship (SKR) study in addition to a traditional SAR analysis at the adenosine A_2A receptor (A_2AR). The ensemble of 24 A_2AR compounds, all triazolotriazine derivatives resembling the prototypic antagonist ZM241385 (4‐(2‐((7‐amino‐2‐(furan‐2‐yl)‐[1,2,4]triazolo[1,5‐a][1,3,5]triazin‐5‐yl)amino)ethyl)phenol), displayed only minor differences in affinity, although they varied substantially in their dissociation rates from the receptor. We believe that such a combination of SKR and SAR analyses, as we have done with the A_2AR, will have general importance for the superfamily of G protein‐coupled receptors, as it can serve as a new strategy to tailor the interaction between ligand and receptor.     

Methamphetamine Blocks Adenosine A2A Receptor Activation via Sigma 1 and Cannabinoid CB1 Receptors

Methamphetamine is, worldwide, one of the most consumed drugs of abuse. One important side effect is neurodegeneration leading to a decrease in life expectancy. The aim of this paper was to check whether the drug affects one of the receptors involved in neurodegeneration/neuroprotection events, namely the adenosine A_2A receptor (A_2AR). First, we noticed that methamphetamine does not affect A_2A functionality if the receptor is expressed in a heterologous system. However, A_2AR becomes sensitive to the drug upon complexes formation with the cannabinoid CB₁ receptor (CB₁R) and the sigma 1 receptor (σ₁R). Signaling via both adenosine A_2AR and cannabinoid CB₁R was affected by methamphetamine in cells co-expressing the two receptors. In striatal primary cultures, the A_2AR–CB₁R heteromer complex was detected and methamphetamine not only altered its expression but completely blocked the A_2AR- and the CB₁R-mediated activation of the mitogen activated protein kinase (MAPK) pathway. In conclusion, methamphetamine, with the participation of σ₁R, alters the expression and function of two interacting receptors, A_2AR, which is a therapeutic target for neuroprotection, and CB₁R, which is the most abundant G protein-coupled receptor (GPCR) in the brain.     

Non-Invasive Assessment of Locally Overexpressed Human Adenosine 2A Receptors in the Heart of Transgenic Mice

A_2A adenosine receptors (A_2A-AR) have a cardio-protective function upon ischemia and reperfusion, but on the other hand, their stimulation could lead to arrhythmias. Our aim was to investigate the potential use of the PET radiotracer [¹⁸F]FLUDA to non-invasively determine the A_2A-AR availability for diagnosis of the A_2AR status. Therefore, we compared mice with cardiomyocyte-specific overexpression of the human A_2A-AR (A_2A-AR TG) with the respective wild type (WT). We determined: (1) the functional impact of the selective A_2AR ligand FLUDA on the contractile function of atrial mouse samples, (2) the binding parameters (B_{max and} K_D) of [¹⁸F]FLUDA on mouse and human atrial tissue samples by autoradiographic studies, and (3) investigated the in vivo uptake of the radiotracer by dynamic PET imaging in A_2A-AR TG and WT. After A_2A-AR stimulation by the A_2A-AR agonist CGS 21680 in isolated atrial preparations, antagonistic effects of FLUDA were found in A_2A-AR-TG animals but not in WT. Radiolabelled [¹⁸F]FLUDA exhibited a K_D of 5.9 ± 1.6 nM and a B_max of 455 ± 78 fmol/mg protein in cardiac samples of A_2A-AR TG, whereas in WT, as well as in human atrial preparations, only low specific binding was found. Dynamic PET studies revealed a significantly higher initial uptake of [¹⁸F]FLUDA into the myocardium of A_2A-AR TG compared to WT. The hA_2A-AR-specific binding of [¹⁸F]FLUDA in vivo was verified by pre-administration of the highly affine A_2AAR-specific antagonist istradefylline. Conclusion: [¹⁸F]FLUDA is a promising PET probe for the non-invasive assessment of the A_2A-AR as a marker for pathologies linked to an increased A_2A-AR density in the heart, as shown in patients with heart failure.     

Effect of Linker Length and Composition on Heterobivalent Ligand‐Mediated Receptor Cross‐Talk between the A1 Adenosine and β2 Adrenergic Receptors

Heterobivalent ligands that possess pharmacophores designed to interact with both the A₁ adenosine receptor (A₁AR) and the β₂ adrenergic receptor (β₂AR) were prepared. More specifically, these ligands contain an adenosine moiety that is linked via its N⁶‐position to the amino group of the saligenin‐substituted ethanolamine moiety present in the well‐known β₂AR agonist, salbutamol. The affinities of these ligands were determined at both receptors and found to vary with linker length and composition. With all compounds, affinity and functional potencies were found to have selectivity for the A₁AR over the β₂AR. In all cases, cAMP accumulation (a β₂AR‐mediated response) was mainly observed when the A₁AR was blocked or its function decreased by pertussis toxin or chronic agonist treatment. This suggests that heterobivalent compounds for receptors that mediate opposite responses might be useful for elucidating the mechanisms of receptor cross‐talk and how this interaction, in terms of responsiveness, may change under pathophysiological conditions.     

Guanosine-Mediated Anxiolytic-Like Effect: Interplay with Adenosine A1 and A2A Receptors

Acute or chronic administration of guanosine (GUO) induces anxiolytic-like effects, for which the adenosine (ADO) system involvement has been postulated yet without a direct experimental evidence. Thus, we aimed to investigate whether adenosine receptors (ARs) are involved in the GUO-mediated anxiolytic-like effect, evaluated by three anxiety-related paradigms in rats. First, we confirmed that acute treatment with GUO exerts an anxiolytic-like effect. Subsequently, we investigated the effects of pretreatment with ADO or A₁R (CPA, CCPA) or A_2AR (CGS21680) agonists 10 min prior to GUO on a GUO-induced anxiolytic-like effect. All the combined treatments blocked the GUO anxiolytic-like effect, whereas when administered alone, each compound was ineffective as compared to the control group. Interestingly, the pretreatment with nonselective antagonist caffeine or selective A₁R (DPCPX) or A_2AR (ZM241385) antagonists did not modify the GUO-induced anxiolytic-like effect. Finally, binding assay performed in hippocampal membranes showed that [³H]GUO binding became saturable at 100–300 nM, suggesting the existence of a putative GUO binding site. In competition experiments, ADO showed a potency order similar to GUO in displacing [³H]GUO binding, whereas AR selective agonists, CPA and CGS21680, partially displaced [³H]GUO binding, but the sum of the two effects was able to displace [³H]GUO binding to the same extent of ADO alone. Overall, our results strengthen previous data supporting GUO-mediated anxiolytic-like effects, add new evidence that these effects are blocked by A₁R and A_2AR agonists and pave, although they do not elucidate the mechanism of GUO and ADO receptor interaction, for a better characterization of GUO binding sites in ARs.     

Design and synthesis of novel dual-action compounds targeting the adenosine A(2A) receptor and adenosine transporter for neuroprotection

A novel compound, N⁶‐(4‐hydroxybenzyl)adenosine, isolated from Gastrodia elata and which has been shown to be a potential therapeutic agent for preventing and treating neurodegenerative disease, was found to target both the adenosine A_2A receptor (A_2AR) and the equilibrative nucleoside transporter 1 (ENT1). As A_2AR and ENT1 are proximal in the synaptic crevice of striatum, where the mutant huntingtin aggregate is located, the dual‐action compounds that concomitantly target these two membrane proteins may be beneficial for the therapy of Huntington’s disease. To design the desired dual‐action compounds, pharmacophore models of the A_2AR agonists and the ENT1 inhibitors were constructed. Accordingly, potentially active compounds were designed and synthesized by chemical modification of adenosine, particularly at the N⁶ and C^5’ positions, if the predicted activity was within an acceptable range. Indeed, some of the designed compounds exhibit significant dual‐action properties toward both A_2AR and ENT1. Both pharmacophore models exhibit good statistical correlation between predicted and measured activities. In agreement with competitive ligand binding assay results, these compounds also prevent apoptosis in serum‐deprived PC12 cells, rendering a crucial function in neuroprotection and potential utility in the treatment of neurodegenerative diseases.     

Synthesis of Novel Pyrido[1,2-c]pyrimidine Derivatives with 6-Fluoro-3-(4-piperidynyl)-1,2-benzisoxazole Moiety as Potential SSRI and 5-HT1A Receptor Ligands

Two series of novel 4-aryl-2H-pyrido[1,2-c]pyrimidine (6a–i) and 4-aryl-5,6,7,8-tetrahydropyrido[1,2-c]pyrimidine (7a–i) derivatives were synthesized. The chemical structures of the new compounds were confirmed by ¹H and ¹³C NMR spectroscopy and ESI-HRMS spectrometry. The affinities of all compounds for the 5-HT_1A receptor and serotonin transporter protein (SERT) were determined by in vitro radioligand binding assays. The test compounds demonstrated very high binding affinities for the 5-HT_1A receptor of all derivatives in the series (6a–i and 7a–i) and generally low binding affinities for the SERT protein, with the exception of compounds 6a and 7g. Extended affinity tests for the receptors D₂, 5-HT_2A, 5-HT₆ and 5-HT₇ were conducted with regard to selected compounds (6a, 7g, 6d and 7i). All four compounds demonstrated very high affinities for the D₂ and 5-HT_2A receptors. Compounds 6a and 7g also had high affinities for 5-HT₇, while 6d and 7i held moderate affinities for this receptor. Compounds 6a and 7g were also tested in vivo to identify their functional activity profiles with regard to the 5-HT_1A receptor, with 6a demonstrating the activity profile of a presynaptic agonist. Metabolic stability tests were also conducted for 6a and 6d.     

Targeting Adenosine Receptors: A Potential Pharmacological Avenue for Acute and Chronic Pain

Adenosine is a purine nucleoside, responsible for the regulation of multiple physiological and pathological cellular and tissue functions by activation of four G protein-coupled receptors (GPCR), namely A₁, A_2A, A_2B, and A₃ adenosine receptors (ARs). In recent years, extensive progress has been made to elucidate the role of adenosine in pain regulation. Most of the antinociceptive effects of adenosine are dependent upon A₁AR activation located at peripheral, spinal, and supraspinal sites. The role of A_2AAR and A_2BAR is more controversial since their activation has both pro- and anti-nociceptive effects. A₃AR agonists are emerging as promising candidates for neuropathic pain. Although their therapeutic potential has been demonstrated in diverse preclinical studies, no AR ligands have so far reached the market. To date, novel pharmacological approaches such as adenosine regulating agents and allosteric modulators have been proposed to improve efficacy and limit side effects enhancing the effect of endogenous adenosine. This review aims to provide an overview of the therapeutic potential of ligands interacting with ARs and the adenosinergic system for the treatment of acute and chronic pain.     

Channel Mimetic Sensing Membranes for Nucleotides Based on Multitopic Hydrogen Bonding

Nucleotide-induced permeability changes of oriented monolayers composed of the nucleotide receptors 4-amino-1-octadecyl-2-(1H)-pyrimidinone ( 1 ), 5-methyl-1-octadecyl-2,4(1H,3H)-pyrimidinedione ( 2 ), and 1-(2-heptylundecyl)-4-(8-(3-methylureido)-2-naphthyl)-amide-2-(1H)-pyrimidinone ( 3 ) were examined. These molecules are capable of binding guanosine, adenosine, and guanosine nucleotides, respectively, by multitopic hydrogen bonding. Monolayers were first formed at the air/water interface and then directly contacted with a highly oriented pyrolytic graphite (HOPG) electrode. The permeabilities of these membranes were evaluated with cyclic voltammetry, using [Fe(CN)₆]^4– as permeability marker. Selectively 5′-GMP-induced decreases in the permeability of the membranes based on receptor 1 or 3 were observed. On the other hand, decreases in the permeability of membranes based on receptor 2 were larger in the presence of 5′-AMP than of 5′-GMP. The permeability changes thus observed can be ascribed to repulsive electrostatic interaction between the marker anions and the negatively charged nucleotides that selectively bind to the electrically neutral membrane receptors. The ratios of the oxidation current decreases for solutions containing 3.0 mM 5′-GMP and for solutions containing 3.0 mM 5′-AMP were 1.30, 0.76, and 1.85 for monolayers based on receptors 1, 2 , or 3 , respectively. The monolayer based on receptor 3 , which is capable of binding the guanine base by five hydrogen bonds, showed a higher 5′-GMP selectivity than the monolayer of receptor 1 with the cytosine residue. Similar nucleotide-induced permeability changes were observed for mixed multilayers of receptor 1 and octadecanol ( 4 ), as well as for mixed monolayers with receptor 2 and 4.     

Insight into the Interactions between Novel Coumarin Derivatives and Human A3 Adenosine Receptors

A study focused on the discovery of new chemical entities based on the 3‐arylcoumarin scaffold was performed with the aim of finding new adenosine receptor (AR) ligands. Thirteen synthesized compounds were evaluated by radioligand binding (A₁, A_2A, and A₃) and adenylyl cyclase activity (A_2B) assays in order to study their affinity for the four human AR (hAR) subtypes. Seven of the studied compounds proved to be selective A₃AR ligands, with 3‐(4′‐methylphenyl)‐8‐(2‐oxopropoxy)coumarin ( 12 ) being the most potent (K_i=634 nM ). None of the compounds showed affinity for the A_2B receptor, while four compounds were found to be nonselective AR ligands for the other three subtypes. Docking simulations were carried out to identify the hypothetical binding mode and to rationalize the interaction of these types of coumarin derivatives with the binding site of the three ARs to which binding was observed. The results allowed us to conclude that the 3‐arylcoumarin scaffold composes a novel and promising class of A₃AR ligands. ADME properties were also calculated, with the results suggesting that these compounds are promising leads for the identification of new drug candidates.     

State-Targeting Stabilization of Adenosine A2A Receptor by Fusing a Custom-Made De Novo Designed α-Helical Protein

G-protein coupled receptors (GPCRs) are known for their low stability and large conformational changes upon transitions between multiple states. A widely used method for stabilizing these receptors is to make chimeric receptors by fusing soluble proteins (i.e., fusion partner proteins) into the intracellular loop 3 (ICL3) connecting the transmembrane helices 5 and 6 (TM5 and TM6). However, this fusion approach requires experimental trial and error to identify appropriate soluble proteins, residue positions, and linker lengths for making the fusion. Moreover, this approach has not provided state-targeting stabilization of GPCRs. Here, to rationally stabilize a class A GPCR, adenosine A_2A receptor (A_2AR) in a target state, we carried out the custom-made de novo design of α-helical fusion partner proteins, which can fix the conformation of TM5 and TM6 to that in an inactive state of A_2AR through straight helical connections without any kinks or intervening loops. The chimeric A_2AR fused with one of the designs (FiX1) exhibited increased thermal stability. Moreover, compared with the wild type, the binding affinity of the chimera against the agonist NECA was significantly decreased, whereas that against the inverse agonist ZM241385 was similar, indicating that the inactive state was selectively stabilized. Our strategy contributes to the rational state-targeting stabilization of GPCRs.     

Several Polyphosphate Kinase 2 Enzymes Catalyse the Production of Adenosine 5′-Polyphosphates

Polyphosphate kinases (PPKs) are involved in many metabolic processes; enzymes of the second family (PPK2) are responsible for nucleotide synthesis fuelled by the consumption of inorganic polyphosphate. They catalyse the phosphorylation of nucleotides with various numbers of phosphate residues, such as monophosphates or diphosphates. Hence, these enzymes are promising candidates for cofactor regeneration systems. Besides adenosine 5′-triphosphate, PPK2s also catalyse the synthesis of highly phosphorylated nucleotides in vitro, as shown here for adenosine 5′-tetraphosphate and adenosine 5′-pentaphosphate. These unusually phosphorylated adenosine 5′-polyphosphates add up to 50 % of the whole adenosine nucleotides in the assay. The two new products were chemically synthesised to serve as standards and compared with the two enzymatically produced compounds by high-performance ion chromatography and ³¹P NMR analysis. This study shows that PPK2s are highly suitable for biocatalytic synthesis of different phosphorylated nucleotides.     

An Fc–Small Molecule Conjugate for Targeted Inhibition of the Adenosine 2A Receptor

The adenosine A_2A receptor (A_2AR) is expressed in immune cells, as well as brain and heart tissue, and has been intensively studied as a therapeutic target for multiple disease indications. Inhibitors of the A_2AR have the potential for stimulating immune response, which could be valuable for cancer immune surveillance and mounting a response against pathogens. One well‐established potent and selective small molecule A_2AR antagonist, ZM‐241385 (ZM), has a short pharmacokinetic half‐life and the potential for systemic toxicity due to A_2AR effects in the brain and the heart. In this study, we designed an analogue of ZM and tethered it to the Fc domain of the immunoglobulin IgG3 by using expressed protein ligation. The resulting protein–small molecule conjugate, Fc–ZM, retained high affinity for two Fc receptors: FcγRI and the neonatal Fc receptor, FcRn. In addition, Fc–ZM was a potent A_2AR antagonist, as measured by a cell‐based cAMP assay. Cell‐based assays also revealed that Fc–ZM could stimulate interferon γ production in splenocytes in a fashion that was dependent on the presence of A_2AR. We found that Fc–ZM, compared with the small molecule ZM, was a superior A_2AR antagonist in mice, consistent with the possibility that Fc attachment can improve pharmacokinetic and/or pharmacodynamic properties of the small molecule.     

2‐Amino[1,2,4]triazolo[1,5‐c]quinazolines and Derived Novel Heterocycles: Syntheses and Structure–Activity Relationships of Potent Adenosine Receptor Antagonists

2‐Amino[1,2,4]triazolo[1,5‐c]quinazolines were identified as potent adenosine receptor (AR) antagonists. Synthetic strategies were devised to gain access to a broad range of derivatives including novel polyheterocyclic compounds. Potent and selective A₃AR antagonists were discovered, including 3,5‐diphenyl[1,2,4]triazolo[4,3‐c]quinazoline ( 17 , K_i human A₃AR 1.16 nm ) and 5′‐phenyl‐1,2‐dihydro‐3′H‐spiro[indole‐3,2′‐[1,2,4]triazolo[1,5‐c]quinazolin]‐2‐one ( 20 , K_i human A₃AR 6.94 nm ). In addition, multitarget antagonists were obtained, such as the dual A₁/A₃ antagonist 2,5‐diphenyl[1,2,4]triazolo[1,5‐c]quinazoline ( 13 b , K_i human A₁AR 51.6 nm , human A₃AR 11.1 nm ), and the balanced pan‐AR antagonists 5‐(2‐thienyl)[1,2,4]triazolo[1,5‐c]quinazolin‐2‐amine ( 11 c , K_i human A₁AR 131 nm , A_2AAR 32.7 nm , A_2BAR 150 nm , A₃AR 47.5 nm ) and 9‐bromo‐5‐phenyl[1,2,4]triazolo[1,5‐c]quinazolin‐2‐amine ( 11 q , K_i human A₁AR 67.7 nm , A_2AAR 13.6 nm , A_2BAR 75.0 nm , A₃AR 703 nm ). In many cases, significantly different affinities for human and rat receptors were observed, which emphasizes the need for caution in extrapolating conclusions between different species.     

Repurposing Dipyridamole in Niemann Pick Type C Disease: A Proof of Concept Study

Niemann Pick type C disease (NPC) is a rare disorder characterized by lysosomal lipid accumulation that damages peripheral organs and the central nervous system. Currently, only miglustat is authorized for NPC treatment in Europe, and thus the identification of new therapies is necessary. The hypothesis addressed in this study is that increasing adenosine levels may represent a new therapeutic approach for NPC. In fact, a reduced level of adenosine has been shown in the brain of animal models of NPC; moreover, the compound T1-11, which is able to weakly stimulate A_2A receptor and to increase adenosine levels by blocking the equilibrative nucleoside transporter ENT1, significantly ameliorated the pathological phenotype and extended the survival in a mouse model of the disease. To test our hypothesis, fibroblasts from NPC1 patients were treated with dipyridamole, a clinically-approved drug with inhibitory activity towards ENT1. Dipyridamole significantly reduced cholesterol accumulation in fibroblasts and rescued mitochondrial deficits; the mechanism elicited by dipyridamole relies on activation of the adenosine A_2AR subtype subsequent to the increased levels of extracellular adenosine due to the inhibition of ENT1. In conclusion, our results provide the proof of concept that targeting adenosine tone could be beneficial in NPC.     

Facile synthesis and characterization of hyperbranched poly(aryl ether ketone)s obtained via an A2 + BB′2 approach

A fast and highly efficient approach for the synthesis of hyperbranched poly(aryl ether ketone)s (HPAEKs) via the polycondensation of A₂ and BB′₂ monomers is described. Commercially available hydroquinone (HQ, A₂ monomer) and easily synthesized 2,4′,6‐trifluorobenzophenone (TF, BB′₂ monomer) were thermally polycondensed to prepared fluoro‐ or phenolic‐terminated HPAEKs with K₂CO₃ and Na₂CO₃ as catalysts. During the reaction, the fluorine at the 4′‐position of TF reacts rapidly with the phenolic group of HQ, forming predominantly dimers and some other species. The dimer can be considered as a new AB′₂ monomer. Further reactions among molecules AB′₂ and AB′₂ with some other species result in the formation of HPAEKs. Fourier transform infrared and ¹H NMR spectra revealed the structures of the resultant polymers. The degree of branching (DB) of the fluoro‐terminated hyperbranched polymers was determined to be in the range 50–57% from ¹H NMR spectra, whereas the DB of the phenolic‐terminated hyperbranched polymers was determined to be 100%. These hyperbranched polymers exhibit excellent solubility in general organic solvents and possess moderate molecular weights with broad distributions determined using gel permeation chromatography. Moreover, the structure and performance of the HPAEKs can be conveniently regulated by adjusting the type and feed ratio of the two monomers. Copyright © 2010 Society of Chemical Industry