The effect of various types of allosteric regulators on basal and hormone-stimulated thyrotropin receptor activity in vitro and in vivo

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Abstract

Background: Long-term antithyroid therapy used to treat autoimmune hyperthyroidism can potentially induce thyroid hormone deficiency. Therefore, the development of thyrotropin receptor regulators which can inhibit thyrotropin activation through orthosteric agonists (thyrotropin, antithyroid autoantibodies) and stimulate its basal activity is crucial. Allosteric thyrotropin regulators with the activity of agonists and negative allosteric modulators has been demonstrated to have this effect.

Aim: To compare the effects of TPY4, a thieno[2,3-d]pyrimidine derivative, and 612-627-Lys(Palm)Ala, a peptide derivative of the third cytoplasmic loop of thyropropin receptor, on the basal and thyropropin-stimulated activity of adenylate cyclase in thyroid membranes and on the basal and thyrotropin-releasing hormone-stimulated levels of thyroid hormones in male rats.

Methods: The activity of adenylate cyclase in plasma membranes of thyroid cells purified from rat thyroids was measured by radioisotope assay using [α-32P]-adenosine triphosphate, radiolabeled sodium adenosine triphosphate, as a substrate. The effect of pharmaceutical agents on basal and thyrotropin-releasing hormone-stimulated levels of thyroid hormone was evaluated by injecting 612–627-Lys(Palm)Ala peptide (at 750 μg/kg) and TPY4 (at 15 mg/kg) to rats. Blood levels of free thyroxine, total triiodothyronine, and thyrotropin were determined by enzyme immunoassay.

Results: Micromolar levels (10–9 M) of TPY4 and 612–627-Lys(Palm)Ala increased the basal activity of adenylate cyclase and decreased thyrotropin stimulation in plasma membranes of thyroid cells. TPY4 demonstrated superior inhibitory activity on thyropropin, whereas the peptide induced a more significant stimulatory response in adenylate cyclase. When administered to rats, both compounds increased the synthesis of free thyroxine and total triiodothyronine, albeit to varying levels. However, they also inhibited the synthesis by thyrotropin-releasing hormone-induced thyroid axis activation (at 100 μg/rat). These compounds did not demonstrate a substantial effect on blood levels of thyrotropin-releasing hormone-stimulated thyropropin, suggesting that they are targeted to thyrotropin-releasing hormone in thyrocytes.

Conclusion: Structurally different allosteric ligands of thyrotropin receptor, i.e., thieno[2,3-d]pyrimidine derivative TPY4 and peptide 612–627-Lys(Palm)Ala, were found to have a similar spectrum of pharmacological activity, functioning as thyrotropin receptor agonists and negative allosteric modulators. Their inhibitory effect on thyrotropin receptor hyperactivation while maintaining euthyroidism offers a promising avenue for the development of novel pharmaceutical agents for the treatment of autoimmune hyperthyroidism.

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About the authors

Kira V. Derkach

Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences

Email: derkatch_k@list.ru
ORCID iD: 0000-0001-6555-9540
SPIN-code: 6925-1558

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Elena A. Shpakova

Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences

Email: eshpakova@mail.ru
ORCID iD: 0000-0002-5645-455X
Russian Federation, Saint Petersburg

Egor A. Didenko

Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences; Saint Petersburg State University

Email: didenkoegor58@mail.ru
ORCID iD: 0009-0000-5217-0624
SPIN-code: 5115-8389
Russian Federation, Saint Petersburg; Saint Petersburg

Viktor N. Sorokoumov

Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences; Saint Petersburg State University

Email: sorokoumov@gmail.com
ORCID iD: 0000-0002-4917-2175
SPIN-code: 1042-8142

Cand. Sci. (Chemistry)

Russian Federation, Saint Petersburg; Saint Petersburg

Alexander O. Shpakov

Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences

Author for correspondence.
Email: alex_shpakov@list.ru
ORCID iD: 0000-0002-4293-3162
SPIN-code: 6335-8311

Dr. Sci. (Biology)

Russian Federation, Saint Petersburg

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