Sekido T, Sakura N, Higashi Y, Miya K, Nitta Y, Nomura M, Sawanishi H, Morito K, Masamune Y, Kasugai S, Yokogawa K, Miyamoto K
Department of Biosynthetic Chemistry Faculty of Pharmaceutical Science, Horuriku University, Kanazawa, Japan
J Drug Target. 2001 Apr;9(2):111-21. doi: 10.3109/10611860108997922.
We synthesized fifteen oligopeptides consisting of Asp or Glu conjugated with a fluorescent probe, 9- fluorenylmethylchloroformate (Fmoc). In the in vitro binding assay to putative hydroxyapatite (HA), the affinities of these conjugates depended only on the number of amino acid residues, not on their optical characters (L or D) or their species (Asp or Glu). In an in vivo experiment involving a single i.v. injection of Fmoc-D-Asp oligopeptides into mice, peptides consisting of over six Asp residues were selectively distributed to the bone. Then, we synthesized estradiol-17 beta-succinate-(L-Asp)6 [E2-(L-Asp)6] and studied its pharmacokinetic characteristics and its antiosteoporotic effects on ovariectomized (OVX) mice. Although the distribution volume of E2-(L-Asp)6 was significantly smaller than that of E2, E2-(L-Asp)6 was selectively distributed in the bone after i.v. injection and gradually decreased during 7 days. E2-(L-Asp)6 effectively prevented OVX-induced bone loss, without altering the uterine weight, in the dosage range of 0.11 to 1.1 mumol/kg once a week, while E2 increased both the bone mineral density and uterine weight at 0.37 mumol/kg every third day. The results suggest that acidic oligopeptide may be useful for drug delivery to bone and E2-(L-Asp)6 is a good candidate as an anti-osteoporosis drug without the adverse side effects of E2.
我们合成了15种由天冬氨酸（Asp）或谷氨酸（Glu）与荧光探针9-芴甲基氯甲酸酯（Fmoc）偶联而成的寡肽。在对假定的羟基磷灰石（HA）进行的体外结合试验中，这些偶联物的亲和力仅取决于氨基酸残基的数量，而不取决于它们的光学性质（L型或D型）或种类（Asp或Glu）。在一项体内实验中，将Fmoc-D-Asp寡肽单次静脉注射到小鼠体内，由超过6个Asp残基组成的肽选择性地分布到骨骼中。然后，我们合成了雌二醇-17β-琥珀酸酯-（L-Asp）6 [E2-（L-Asp）6]，并研究了其药代动力学特征及其对去卵巢（OVX）小鼠的抗骨质疏松作用。尽管E2-（L-Asp）6的分布容积明显小于E2，但静脉注射后E2-（L-Asp）6选择性地分布在骨骼中，并在7天内逐渐减少。在每周一次0.11至1.1 μmol/kg的剂量范围内，E2-（L-Asp）6有效预防了OVX诱导的骨质流失，且未改变子宫重量，而E2每三天以0.37 μmol/kg的剂量给药时，增加了骨矿物质密度和子宫重量。结果表明，酸性寡肽可能有助于将药物递送至骨骼，并且E2-（L-Asp）6是一种很好的抗骨质疏松药物候选物，没有E2的不良副作用。
