Growth hormone secretagogue receptor based protein
Ghrelin is a peptide hormone that binds its receptor, growth hormone secretatgogue receptor 1a (GHS-R1 a, ghrelin receptor), to promote adiposity and obesity in mammals. Ghrelin and its receptor are targets for therapeutic intervention to treat obesity-related disease and cancer. A soluble decoy GHS-R1 a receptor is developed that binds ghrelin in the periphery, preventing ghrelin from binding GHS-R1 on cells, thereby antagonizing ghrelin to treat obesity-related pathological conditions and cancer. GHS-R1 a is a transmembrane protein comprising an N-terminal extracellular domain (Nt), seven transmembrane regions and three extracellular loops (EC1, EC2 and EC3). The Nt, EC1 and EC2 are linked together, in the absence of the transmembrane regions, and fused to a Fc from an immunoglobulin, to create the decoy GHS-R1 a fusion protein, GHSR-Fc. The GHSR-Fc inhibits adiposity and weight gain in mice on a high fat diet (HFD), while the Nt and ECs on their own have no significant effect.
1. A soluble fusion molecule comprising
a)
i) the N-terminal of human growth hormone secretagogue receptor (GHS-R1a) having an amino acid sequence as shown in SEQ ID NO: 13 or a variant thereof with at least 95% sequence identity, linked to extracellular loop 1 (EC1) having an amino acid sequence as shown in SEQ ID NO: 15 or a variant thereof with at least 95% sequence identity
ii) the N-terminal of GHS-R1a having an amino acid sequence as shown in SEQ ID NO: 13 or a variant thereof with at least 95% sequence identity, linked to extracellular loop 1 having an amino acid sequence as shown in SEQ ID NO: 15 or a variant thereof with at least 95% sequence identity, or
iii) the N-terminal of GHS-R1a having an amino acid sequence as shown SEQ ID NO: 13 or a variant thereof with at least 95% sequence identity linked to extracellular loop 1 having an amino acid sequence as shown in SEQ ID NO: 15 or a variant thereof with at least 95% sequence identity and extracellular loop 2 having an amino acid sequence as shown SEQ ID NO: 17 or a variant thereof with at least 95% sequence identity; linked to; and
b) a fusion partner that is an immunoglobulin G (IgG) constant region (Fc) of IgG 2 or IgG4.
2. The soluble fusion molecule of claim 1 , wherein a) comprises or consists of i) the N-terminal, ii) the extracellular loop 1 and iii) the extracellular loop 2 of the human GHS-R1a.
3. The soluble fusion molecule of claim 1 , wherein the N-terminal of human GHS-R1a is encoded by the nucleic acid sequence as shown in SEQ ID NO: 12 or a variant thereof with at least 95% sequence identity; wherein the extracellular loop 1 of human GHS R1a is encoded by the nucleic acid sequence as shown in SEQ ID NO: 14 or a variant thereof with at least 95% sequence identity; and/or wherein the extracellular loop 2 of human GHS R1a is encoded by a nucleic acid sequence as shown in SEQ ID NO: 16 or a variant thereof with at least 95% sequence identity.
4. The soluble fusion molecule of claim 1 , wherein the Fc region is the Fc region of IgG2.
5. The soluble fusion molecule of claim 1 , further comprising a peptide linker between a) and b).
6. The soluble fusion molecule of claim 1 comprising the amino acid sequence as shown in SEQ ID NO: 11 or a variant thereof with at least 95% sequence identity or encoded by the nucleic acid sequence as shown in SEQ ID NO: 10 or a variant thereof with at least 95% sequence identity.
7. A nucleic acid molecule encoding the soluble fusion molecule of claim 1 .
8. A host cell comprising the nucleic acid molecule of claim 7 .
9. A pharmaceutical composition comprising the soluble fusion molecule of claim 1 , a nucleic acid molecule encoding said soluble fusion molecule or a host cell expressing said nucleic acid molecule, and a pharmaceutically acceptable carrier.
10. The pharmaceutical composition of claim 9 comprising the soluble fusion molecule and a pharmaceutically acceptable carrier.