MON-459 ONO-5788, a Novel Oral Small Molecule Somatostatin Receptor Type-2 (SST2) Agonist, Attenuates GH Hypersecretion in Human GH-Secreting, Pituitary Adenoma-Derived Cells

Abstract

Somatostatin receptor ligands (SRL) and GHR antagonists are used for persistent or recurrent acromegaly following noncurative surgery and as primary therapy for patients unsuitable for surgery. We tested in vitro action of a novel oral SRL compound ONO-5788 and its active metabolite (ONO-ST1-641) which signals via SST2. With IRB approval and informed consent, 12 human somatotroph adenomas, obtained at transsphenoidal resection were cultured. In the 1st cohort, 8 somatotroph adenomas were treated with ONO-ST1-641 (0, 100, 1000 nM) and octreotide (OCT, 100 nM) for 24 h, and media collected at 2, 6, and 24 h respectively to assess GH and PRL concentrations by ELISA. Cells harvested at 24 h were used to measure mRNA levels of SST2 and SST5, hormone genes and cell cycle genes CDKN1A, CDKN1B by qPCR. Three of 8 tumors (38%) responded to ONO-ST1-641 and OCT as early as 2 and 6 h. GH was significantly reduced by OCT (18% to 58% reduction, P<0.05) and similarly attenuated by ONO-ST1-641 (28 to 59% reduction, P<0.05). GH secretion in 6 of 8 tumors (75%) was decreased at 24 h, showing reduction by OCT and ONO-ST1-641 by up to 48% (P<0.01) and 50% (P<0.01), respectively. PRL levels were undetectable, very low or unchanged by treatments. Related compound ONO-5788 showed similar responses. SST2 and SST5 expression was unchanged during treatments in the 6/8 responders as assessed by TaqMan qPCR using HPRT1 and GAPDH as controls. Neither was GH, PRL, CGA, CDKN1A and CDKN1B mRNA expression altered by ONO-ST1-641 or OCT for 24 h, as assessed by SYBR qPCR using RPL13A, GAPDH and 18S as controls, suggesting that ONO-ST1-641 did not affect these hormone genes and cell cycle regulator mRNA levels. In the 2nd cohort, pretreatment baseline SST2 and SST5 expression were assessed in 4 cultured tumors treated with ONO-ST1-641 (0, 100, 1000 nM) or OCT (100 nM) for 6 h. Untreated tumor cell mRNA was extracted at baseline to assess SST2 and SST5 expression and GH and PRL measured at 6 h. Two of 4 tumors were unresponsive to both ONO-ST1-641 and OCT, 1 showed mild GH reduction by ONO-ST1-641 (15%) but not by OCT. Notably, 1 specimen expressing highly abundant baseline SST2 exhibited marked GH suppression by ONO-ST1-641 and OCT at 6 h (63 and 62%, respectively, P<0.01). High baseline SST2 expression further validated by Western blot and confocal immunofluorescence, imply that SST receptor expression associates with ONO-ST1-641 responsiveness. Available SRLs and GHR antagonists are parenteral, requiring lifelong injection, with variable and suboptimal efficacy, necessitating development of novel oral small molecule drugs for acromegaly. Our study provides preclinical evidence that ONO-5788 and its active metabolite attenuate GH hypersecretion in primary somatotroph adenoma cells cultures, suggesting the potential application of ONO-5788 for acromegaly patients.