BMS-707035

製品コードS1366 バッチS136601

化学情報

	Synonyms	N/A	Storage (From the date of receipt)	3 years -20°C powder 1 years -80°C in solvent
	化学式	C₁₇H₁₉FN₄O₅S
	分子量	410.42	CAS No.	729607-74-3
Solubility (25°C)*	体外	DMSO	38 mg/mL (92.58 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

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生物活性

製品説明	BMS-707035 is a specific HIV-I integrase (IN) inhibitor with IC50 of 15 nM. Phase 2.
in vitro	BMS-707035 is a pyrimidine carboxamide similar to Raltegravir, the first integrase inhibitor licensed for clinical use. BMS-707035 is a potent, specific, and reversible HIV-I integrase (IN) inhibitor that blocks HIV IN strand transfer activity with IC50 of 15 nM. ^[1] However, several IN mutations, including V75I, Q148R, V151I, and G163R are found to confer resistance to HIV IN inhibitors. The binding of BMS-707035 and target DNA to IN are mutually exclusive events, as revealed by the fact that the inhibition of strand transfer catalysis by BMS-707035 is overcome by increasing amount of target DNA. The binding affinity of BMS-707035 to IN is also affected by the four terminal bases at the 5' end of the pre-processed U5 long terminal repeat (LTR). Gln¹⁴⁸ of IN is crucial for the binding of BMS-707035 to IN. ^[1] The 3' terminus of the viral LTR, on the other hand, retards the rate of BMS-707035 association with IN, by regulating the kinetics of binding and dissociation. ^[2]

製品説明

BMS-707035 is a specific HIV-I integrase (IN) inhibitor with IC50 of 15 nM. Phase 2.

in vitro

BMS-707035 is a pyrimidine carboxamide similar to Raltegravir, the first integrase inhibitor licensed for clinical use. BMS-707035 is a potent, specific, and reversible HIV-I integrase (IN) inhibitor that blocks HIV IN strand transfer activity with IC50 of 15 nM. ^[1] However, several IN mutations, including V75I, Q148R, V151I, and G163R are found to confer resistance to HIV IN inhibitors. The binding of BMS-707035 and target DNA to IN are mutually exclusive events, as revealed by the fact that the inhibition of strand transfer catalysis by BMS-707035 is overcome by increasing amount of target DNA. The binding affinity of BMS-707035 to IN is also affected by the four terminal bases at the 5' end of the pre-processed U5 long terminal repeat (LTR). Gln¹⁴⁸ of IN is crucial for the binding of BMS-707035 to IN. ^[1] The 3' terminus of the viral LTR, on the other hand, retards the rate of BMS-707035 association with IN, by regulating the kinetics of binding and dissociation. ^[2]

プロトコル(参考用のみ)

キナーゼアッセイ	Determination of Strand Transfer Activity and Inhibition
キナーゼアッセイ	The in vitro activities of purified INs in combination with the various duplex LTRs are measured through a scintillation proximity assay (SPA). In a first step, the viral LTR duplexes are prepared by annealing individual oligonucleotides. The viral (donor) LTR DNA is then attached, via a 5'-biotin linker on the plus strand, to streptavidin-coated SPA PVT beads as follows. SPA PVT beads (10 mg) are suspended in 0.2 mL of PBS. The suspension is then centrifuged at <5000 × g for 15 min. The supernatant is removed, and the pellet is resuspended with 0.2 mL of PBS, 0.85 M NaCl, and 21 μL of 12 μM duplex HIV LTR DNA. The sequences of the duplexes are as follows, except for the variations in the underlined bases: plus strand, 5'-biotin-ACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA; minus strand, 5'-ACTGCTAGAGATTTTCCACACTGACTAAAAG. The LTR DNA is allowed to bind for 60 min at room temperature with gentle rocking, after which time 0.8 mL of TE is added. The mixture is then centrifuged at <5000 × g and resuspended in 0.8 mL of TE, 50 mM NaCl. The beads are washed 4 additional times with TE, 50 mM NaCl, each time centrifuging to remove unbound viral LTR DNA. The final pellet is resuspended in 0.2 mL of PBS and stored at 4 °C before use. Enzyme complexes for 80 strand transfer reactions are prepared as follows: 0.15 mL of bead-DNA complexes, 2.25 mL of SPA buffer (13.3 mM dithiothreitol, 32 mM MOPS, pH 7.0, 0.067% NP-40, 6.4% polyethylene glycol, 25.6 mM MgCl₂, 12.8% (v/v) Me₂SO, and 100 mM NaCl), and IN (37 μg of WT, 88 μg of N155H, and 36 μg of Q148R) are incubated at 37 °C. After 1.5 hours, complexes are pelleted and resuspended with 2.4 mL of SPA buffer. The proper amount of each IN is determined through titration experiments and represented the minimal amount of enzyme required to produce the maximal amount of strand transfer products. The target DNA is prepared (5'-[³³P]TGACCAAGGGCTAATTCACT-3' annealed to 5'-[³³P]AGTGAATTAGCCCTTGGTCA-3') in a separate step by individually 5' end labeling each of the oligonucleotides with [γ-³³P]ATP. The ³³P-labeled oligonucleotides are then annealed to form the target duplex DNA. Strand transfer assays consists of combining 30 μL of IN complexes with 10 μL of 25% Me₂SO/H₂O) (v/v) in white microtiter plates. After 10 min of incubation at 37 °C, 10 μL of ³³P-labeled target DNA (1 × 10⁶ cpm) is added (final concentration of target is 0.92 nM). Plates are returned to 37 °C for 2 hours, after which time the strand transfer reactions are stopped by the addition of 200 μL of PBS, 50 mM EDTA. Plates are allowed to stand overnight before reading on a Topcount scintillation counter. HIV-1 IN complexes are evaluated for inhibitor sensitivity using the SPA assay, except that 10 μL of a 5-fold serial dilution of BMS-707035 in 25% (v/v) Me₂SO/H₂O is added in the place of 10 μL of 25% (v/v) Me₂SO/H₂O. Data are analyzed by fitting to a sigmoidal dose-response curve.

参考

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

Activation of PERK-ATF4-CHOP pathway as a novel therapeutic approach for efficient elimination of HTLV-1-infected cells. [ Blood Adv, 2020, 12;4(9):1845-1858]	PubMed: 32369565

Activation of PERK-ATF4-CHOP pathway as a novel therapeutic approach for efficient elimination of HTLV-1-infected cells. [ Blood Adv, 2020, 12;4(9):1845-1858]

PubMed: 32369565

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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