AM095 is a potent LPA1 receptor antagonist with IC50 values of 0.98 and 0.73 μM for recombinant human or mouse LPA1 respectively.

LPA1 (recombinant human):0.98 μM, LPA1 (mouse):0.73 μM

AM095是一种强效的LPA1受体拮抗剂，其通过抑制GTPγS与大量表达重组人类或鼠类LPA1的中国仓鼠卵巢(CHO)细胞膜的结合来发挥作用，其IC50值分别为0.98和0.73 μM。此外，AM095能够抑制大量表达鼠LPA1的CHO细胞（IC50=778 nM）和人类A2058黑色素瘤细胞（IC50=233 nM）中由LPA驱动的趋化作用。AM095在人类LPA1 GTPγS结合测定中的IC50与我们之前发表的化合物AM966（IC50=0.98±0.17 μM）和Debio-0719化合物（IC50=0.60±0.04 μM）相当[1]。AM095还能抑制稳定转染有人类或鼠类LPA1的CHO细胞中由LPA引起的钙通量，其对于LPA引起的人类或鼠类LPA1-转染CHO细胞钙通量的拮抗作用IC50值分别为0.025和0.023 μM[2]。

AM095具有高口服生物利用度和中等半衰期，在大鼠和犬类中口服和静脉注射给药时耐受性良好。在大鼠口服给药（10 mg/kg）后，AM095的血浆浓度在2小时达到峰值，Cmax为41 μM，之后在24小时降至10 nM。静脉注射（2 mg/kg）后，15分钟内观察到Cmax为12 μM，24小时后也降至约10 nM，半衰期为1.79小时。在犬类中，单次口服给药5 mg/kg，15分钟内血浆浓度达到峰值21 μM，24小时后降至10 nM。相比之下，静脉注射给药2 mg/kg，15分钟内达到Cmax 11 μM，并在8小时降至15 nM，半衰期为1.5小时[1]。

Known amounts of AM095 (diluted in DMSO) or vehicle (DMSO) are added to 25 to 40 μg of hLPA1/CHO or mLPA1/CHO membranes and 0.1 nM [35S]-GTPγS in buffer (50 mM HEPES, 0.1 mM NaCl, 10 mM MgCl2, 50 μg/mL saponin, pH 7.5) containing 0.2% fatty acid-free human serum albumin and 5 μM GDP. To test for LPA1 antagonist activity, the ability of AM095 to inhibit GTPγS binding stimulated by 900 nM LPA (18:1) is measured. Alternatively, to test for agonist effects, the ability of AM095 to stimulate GTPγS binding in the absence of LPA is measured. Reactions are incubated for 30 min at 30°C, before harvesting membranes onto glass filter binding plates (UniFilter GF/B) and washing three times with cold buffer containing 50 mM HEPES, pH 7.4, 100 mM NaCl, 10 mM MgCl2 using a Brandel 96-tip cell harvester. Plates are dried and then cpm are evaluated by using a Packard TopCount NXT microplate scintillation counter[1].