Crystallization and preliminary analysis of bovine adenosine deaminase

Kinoshita, Takayoshi; Nishio, Nobuya; Sato, Akihiro; Murata, Masayoshi

doi:10.1107/s090744499901183x

Cited by 12 publications

(11 citation statements)

References 17 publications

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“…For all assay screening, bovine spleen ADA was used, as it has 91 % sequence similarity overall with human ADA, and is highly conserved at the active site [15] . Therefore, the values obtained with bovine spleen ADA should be comparable with human ADA.…”

Section: Methodsmentioning

confidence: 99%

Identification of Adenosine Deaminase Inhibitors by Metal‐binding Pharmacophore Screening

et al. 2020

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Adenosine deaminase (ADA) is a human mononuclear Zn 2 + metalloenzyme that converts adenosine to inosine. ADA is a validated drug target for cancer, but there has been little recent work on the development of new therapeutics against this enzyme. The lack of new advancements can be partially attributed to an absence of suitable assays for high-throughput screening (HTS) against ADA. To facilitate more rapid drug discovery efforts for this target, an in vitro assay was developed that utilizes the enzymatic conversion of a visibly emitting adenosine analogue to the corresponding fluorescent inosine analogue by ADA, which can be monitored via fluorescence intensity changes. Utilizing this assay, a library of~350 small molecules containing metal-binding pharmacophores (MBPs) was screened in an HTS format to identify new inhibitor scaffolds against ADA. This approach yielded a new metalbinding scaffold with a K i value of 26 � 1 μM.

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Section: Methodsmentioning

confidence: 99%

Identification of Adenosine Deaminase Inhibitors by Metal‐binding Pharmacophore Screening

et al. 2020

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“…ADA is present in many human tissues, but the highest levels have been found in lymph nodes, spleen and thymus [1]. Bovine ADA is more closely related to human ADA [10,11].…”

Section: Introductionmentioning

confidence: 99%

A study on the inhibition of adenosine deaminase

Alunni

Orrù

Ottavi

2008

Journal of Enzyme Inhibition and Medicinal Chemistry

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Adenosine deaminase (ADA, EC 3.5.4.4) catalyses the irreversible deamination of adenosine and 2 0 -deoxyadenosine to inosine and 2 0 -deoxyinosine, respectively. In this study the inhibition of ADA from bovine spleen by several molecules with structure related to that of the substrate or product has been quantified. The inhibitors adenine, purine, inosine, 2-aminopurine, 4-aminopyrimidine, 4-aminopyridine, 4-hydroxypyridine and phenylhydrazine are shown to be competitive inhibitors with K I (mM) values of 0.17, 1.1, 0.35, 0.33, 1.3, 1.8, 1.4 and 0.25, respectively. Synergistic inhibition by various combinations of molecules that imitate the structure of the substrate has never been observed. Some general conclusions are: i) the enzyme ADA from bovine spleen we have used is appropriate for kinetic studies of inhibition and mechanistic studies; it can be a reference catalytic system for the homogeneous comparison of various inhibitors; ii) this enzyme presents very rigid requirements for binding the substrate: variations in the structure of adenosine imply the loss of important interactions.

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“…The organic layer was dried (Na 2 SO 4 ) and concentrated in vacuo. The residue was purified by silica gel (66 g) column chromatography eluting with chloroform (CHCl 3 )/methanol (MeOH) (30:1) to give 13 (489 mg, 25.4%); IR (KBr, cm -1 ) 3600−3000, 1662, 1504; 1 H NMR (CDCl 3 ) δ −0.08 (3H, s), −0.06 (3H, s), 0.81 (9H, s), 2.25−2.65 (2H, m), 3.65−4.20 (5H, m), 5.44 (1H, brs), 6.62 (1H, d, J = 3.1 Hz), 6.97 (1H, brs), 7.16 (1H, d, J = 3.1 Hz), 7.40 (1H, s), 7.66 (1H, s), 7.68 (1H, d, J = 8.8 Hz), 8.04 (1H, dd, J = 8.8 Hz, 2.0 Hz), 8.18 (1H, d, J = 2.0 Hz); MS (ESI, m / z ) 458 (M + H) + ; [α] D −3.3° ( c 0.50, EtOH).…”

Section: Methodsmentioning

confidence: 99%

“…Crystallography of 4e/ADA. Cocrystals of the bovine intestine ADA were obtained using procedures similar to those previously described . In this study, 100 mM MES, pH 6.0, 2.1 M ammonium sulfate, and 2.5% (v/v) PEG-400 were used as precipitant.…”

Section: Methodsmentioning

confidence: 99%

Structure-Based Design, Synthesis, and Structure−Activity Relationship Studies of Novel Non-nucleoside Adenosine Deaminase Inhibitors

Terasaka

Kinoshita²,

Kuno³

et al. 2004

J. Med. Chem.

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We disclose herein optimization efforts around the novel, highly potent non-nucleoside adenosine deaminase (ADA) inhibitor, 1-[(R)-1-hydroxy-4-(6-(3-(1-methylbenzimidazol-2-yl)propionylamino)indol-1-yl)-2-butyl]imidazole-4-carboxamide 1 (K(i)= 7.7 nM), which we recently reported. Structure-based drug design (SBDD) utilizing the crystal structure of the 1/ADA complex was performed in order to obtain structure-activity relationships (SAR) for this type of compound rationally and effectively. To utilize the newly formed hydrophobic space (F2), replacement of the benzimidazole ring of 1 with a n-propyl chain (4b) or a simple phenyl ring (4c) was tolerated in terms of binding activity, and the length of the methylene-spacer was shown to be optimal at two or three. Replacement of an amide with an ether as a linker was also well tolerated in terms of binding activity and moreover improved the oral absorption (6a and 6b). Finally, transformation of indol-1-yl to indol-3-yl resulted in discovery of a novel highly potent and orally bioavailable ADA inhibitor, 1-[(R)-4-(5-(3-(4-chlorophenyl)propoxy)-1-methylindol-3-yl)-1-hydroxy-2-butyl]imidazole-4-carboxamide 8c.

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Crystallization and preliminary analysis of bovine adenosine deaminase

Cited by 12 publications

References 17 publications

Identification of Adenosine Deaminase Inhibitors by Metal‐binding Pharmacophore Screening

Identification of Adenosine Deaminase Inhibitors by Metal‐binding Pharmacophore Screening

A study on the inhibition of adenosine deaminase

Structure-Based Design, Synthesis, and Structure−Activity Relationship Studies of Novel Non-nucleoside Adenosine Deaminase Inhibitors

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