Kim Yong-Chul, Camaioni Emidio, Ziganshin Airat U, Ji Xiao-Duo, King Brian F, Wildman Scott S, Rychkov Alexei, Yoburn Joshua, Kim Heaok, Mohanram Arvind, Harden T Kendall, Boyer José L, Burnstock Geoffrey, Jacobson Kenneth A
Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland.
Drug Dev Res. 1998 Oct 1;45(2):52-66. doi: 10.1002/(SICI)1098-2299(199810)45:2<52::AID-DDR2>3.0.CO;2-V.
Novel analogs of the P2 receptor antagonist pyridoxal-5'-phosphate-6-phenylazo-2',4'-disulfonate (PPADS) were synthesized. Modifications were made through functional group substitution on the sulfophenyl ring and at the phosphate moiety through the inclusion of phosphonates, demonstrating that a phosphate linkage is not required for P2 receptor antagonism. Substituted 6-phenylazo and 6-naphthylazo derivatives were also evaluated. Among the 6-phenylazo derivatives, 5'-methyl, ethyl, propyl, vinyl, and allyl phosphonates were included. The compounds were tested as antagonists at turkey erythrocyte and guinea-pig taenia coli P2Y(1) receptors, in guinea-pig vas deferens and bladder P2X(1) receptors, and in ion flux experiments by using recombinant rat P2X(2) receptors expressed in Xenopus oocytes. Competitive binding assay at human P2X(1) receptors in differentiated HL-60 cell membranes was carried out by using [(35)S]ATP-γ-S. A 2'-chloro-5'-sulfo analog of PPADS (C(14)H(12)O(9)N(3)ClPSNa), a vinyl phosphonate derivative (C(15)H(12)O(11)N(3)PS(2)Na(3)), and a naphthylazo derivative (C(18)H(14)O(12)N(3)PS(2)Na(2)), were particularly potent in binding to human P2X(1) receptors. The potencies of phosphate derivatives at P2Y(1) receptors were generally similar to PPADS itself, except for the p-carboxyphenylazo phosphate derivative C(15)H(13)O(8)N(3)PNa and its m-chloro analog C(15)H(12)O(8)N(3)ClPNa, which were selective for P2X vs. P2Y(1) receptors. C(15)H(12)O(8)N(3)ClPNa was very potent at rat P2X(2) receptors with an IC(50) value of 0.82 μM. Among the phosphonate derivatives, [4-formyl-3-hydroxy-2-methyl-6-(2-chloro-5-sulfonylphenylazo)-pyrid-5-yl]methylphosphonic acid (C(14)H(12)-O(8)N(3)ClPSNa) showed high potency at P2Y(1) receptors with an IC(50) of 7.23 μM. The corresponding 2,5-disulfonylphenyl derivative was nearly inactive at turkey erythrocyte P2Y(1) receptors, whereas at recombinant P2X(2) receptors had an IC(50) value of 1.1 μM. An ethyl phosphonate derivative (C(15)H(15)O(11)N(3)PS(2)Na(3)), whereas inactive at turkey erythrocyte P2Y(1) receptors, was particularly potent at recombinant P2X(2) receptors.
合成了P2受体拮抗剂吡哆醛 - 5'-磷酸 - 6 - 苯基偶氮 - 2',4'-二磺酸盐（PPADS）的新型类似物。通过在磺苯基环上进行官能团取代以及通过引入膦酸盐对磷酸部分进行修饰，表明P2受体拮抗作用并不需要磷酸连接。还评估了取代的6 - 苯基偶氮和6 - 萘基偶氮衍生物。在6 - 苯基偶氮衍生物中，包括5'-甲基、乙基、丙基、乙烯基和烯丙基膦酸盐。这些化合物在火鸡红细胞和豚鼠结肠带P2Y(1)受体、豚鼠输精管和膀胱P2X(1)受体上作为拮抗剂进行了测试，并在使用非洲爪蟾卵母细胞中表达的重组大鼠P2X(2)受体的离子通量实验中进行了测试。通过使用[(35)S]ATP - γ - S在分化的HL - 60细胞膜中的人P2X(1)受体上进行了竞争性结合测定。PPADS的2'-氯 - 5'-磺基类似物（C(14)H(12)O(9)N(3)ClPSNa）、乙烯基膦酸盐衍生物（C(15)H(12)O(11)N(3)PS(2)Na(3)）和萘基偶氮衍生物（C(18)H(14)O(12)N(3)PS(2)Na(2)）在与人P2X(1)受体结合方面特别有效。除了对羧基苯基偶氮磷酸盐衍生物C(15)H(13)O(8)N(3)PNa及其间氯类似物C(15)H(12)O(8)N(3)ClPNa对P2X受体相对于P2Y(1)受体具有选择性外，磷酸盐衍生物在P2Y(1)受体上的效力通常与PPADS本身相似。C(15)H(12)O(8)N(3)ClPNa在大鼠P2X(2)受体上非常有效，IC(50)值为0.82μM。在膦酸盐衍生物中，[4 - 甲酰基 - 3 - 羟基 - 2 - 甲基 - 6 - (2 - 氯 - 5 - 磺酰基苯基偶氮) - 吡啶 - 5 - 基]甲基膦酸（C(14)H(12) - O(8)N(3)ClPSNa）在P2Y(1)受体上显示出高效力，IC(50)为7.23μM。相应的2,5 - 二磺酰基苯基衍生物在火鸡红细胞P2Y(1)受体上几乎无活性，而在重组P2X(2)受体上IC(50)值为1.1μM。一种乙基膦酸盐衍生物（C(15)H(15)O(11)N(3)PS(2)Na(3)）在火鸡红细胞P2Y(1)受体上无活性，但在重组P2X(2)受体上特别有效。
