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JP-7855180-B2 - Screening assays, modulators, and regulators for activation of the advanced glycation end product receptor (RAGE).

JP7855180B2JP 7855180 B2JP7855180 B2JP 7855180B2JP-7855180-B2

Inventors

  • プレーガー，ケヴィン・ドナルド・ジョージ
  • トーマス，マーリン・クリストファー
  • ピカリング，レイリーン・ジェイン
  • ロサド，カルロス
  • ティケリス，クリストス

Assignees

  • モナシュ・ユニヴァーシティ
  • ザ・ユニヴァーシティ・オヴ・ウェスタン・オーストラリア

Dates

Publication Date
20260508
Application Date
20230726
Priority Date
20170822

Claims (20)

  1. An isolated or purified peptide that inhibits RAGE ligand-independent signaling, wherein the peptide is An isolated or purified peptide having an amino acid sequence that is at least 90% identical to the continuous amino acid sequence of residues 390 to at least 379 of SEQ ID NO: 14, wherein the C-terminus of the peptide corresponds to residue 390 of SEQ ID NO: 14, and the amino acid corresponding to residue 379 of SEQ ID NO: 14 is Q or a conserved amino acid substitution thereof.
  2. The isolated or purified peptide according to claim 1, comprising an amino acid sequence that is at least 90% identical to the continuous amino acid sequence from residue 390 to at least 379 of SEQ ID NO: 14, wherein the C-terminus of the peptide corresponds to residue 390 of SEQ ID NO: 14, and the amino acid corresponding to residue 379 of SEQ ID NO: 14 is Q, K, R, D, N, or E.
  3. (i) An amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 22 to 390 of SEQ ID NO: 14; (ii) An amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 370 to 390 of SEQ ID NO: 14; (iii) An amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 370 to 390 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 379 of SEQ ID NO: 14 is K; (iv) An amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 370 to 390 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 388 of SEQ ID NO: 14 is A; (v) An amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 370 to 390 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 380 of SEQ ID NO: 14 is A; (vi) an amino acid sequence that is at least 90% identical to the continuous amino acid sequence consisting of residues 370 to 390 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 382 of SEQ ID NO: 14 is A; (vii) an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of residues 370 to 390 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 384 of SEQ ID NO: 14 is A; or (viiii) an isolated or purified peptide according to claim 1, comprising an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of residues 374 to 390 of SEQ ID NO: 14.
  4. The isolated or purified peptide according to claim 1, wherein the amino acid at the position corresponding to residue 388 of SEQ ID NO: 14 is not alanine.
  5. The isolated or purified peptide according to claim 1, wherein the amino acid at the position corresponding to residue 388 of SEQ ID NO: 14 is leucine.
  6. An isolated or purified peptide that inhibits RAGE ligand-independent signaling, wherein the peptide is (i) an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of SEQ ID NO: 14, and includes at least both residues 379 and 391 of SEQ ID NO: 14, The amino acid at the position corresponding to residue 379 of SEQ ID NO: 14 is Q or its conserved amino acid substitution, and (ii) an amino acid sequence in which the amino acid at the position corresponding to residue 391 of SEQ ID NO: 14 is not S; or (ii) an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of SEQ ID NO: 14, including at least both residues 379 and 391 of SEQ ID NO: 14 The amino acid at the position corresponding to residue 379 of sequence number 14 is not Q, and An isolated or purified peptide consisting of a sequence in which the amino acid at the position corresponding to residue 391 of SEQ ID NO: 14 is not S.
  7. The aforementioned peptide An amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of SEQ ID NO: 14, and includes at least both residues 379 and 391 of SEQ ID NO: 14, The amino acid at the position corresponding to residue 379 of sequence number 14 is Q, K, R, D, N, or E, and The isolated or purified peptide according to claim 6, comprising a sequence in which the amino acid at the position corresponding to residue 391 of SEQ ID NO: 14 is Y, C, V, R, N, K, H, G, F, E, D, or A.
  8. The isolated or purified peptide according to claim 6, wherein the amino acid at the position corresponding to residue 391 of SEQ ID NO: 14 is A.
  9. The aforementioned peptide (i) an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence consisting of residues 362 to 404 of Sequence ID No. 14, wherein the amino acid at the position corresponding to residue 391 is Y, C, V, R, N, K, H, G, F, E, D, or A; (ii) an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of residues 22 to 404 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 391 is A, C, or E; or (iii) an isolated or purified peptide according to claim 6, comprising an amino acid sequence that is at least 90% identical to the consecutive amino acid sequence of residues 342 to 404 of SEQ ID NO: 14, wherein the amino acid at the position corresponding to residue 391 of SEQ ID NO: 14 is A.
  10. The isolated or purified peptide according to claim 6, wherein the amino acid at the position corresponding to residue 388 of SEQ ID NO: 14 is not alanine.
  11. The isolated or purified peptide according to claim 6, wherein the amino acid at the position corresponding to residue 388 of SEQ ID NO: 14 is leucine.
  12. An isolated or purified peptide according to any one of claims 1 to 11 , (i) A reporter component selected from the group consisting of enzymes, luminescent or bioluminescent molecules, and fluorescent molecules, or (ii) Peptides used for affinity purification, peptides that guide fusion polypeptides into intracellular compartments of eukaryotic cells, or peptides that promote translocation of eukaryotic cell membranes. A fusion polypeptide consisting of and .
  13. A nucleic acid comprising a nucleotide sequence encoding an isolated or purified peptide according to any one of claims 1 to 11.
  14. A nucleic acid comprising a nucleotide sequence encoding the fusion polypeptide described in claim 12.
  15. A pharmaceutical composition comprising a therapeutically effective amount of an isolated or purified peptide according to any one of claims 1 to 11.
  16. A pharmaceutical composition comprising a therapeutically effective amount of the fusion polypeptide described in claim 12.
  17. A pharmaceutical composition comprising a therapeutically effective amount of the nucleic acid described in claim 13.
  18. A pharmaceutical composition comprising a therapeutically effective amount of the nucleic acid described in claim 14.
  19. A pharmaceutical composition comprising a therapeutically effective amount of an isolated or purified peptide according to any one of claims 1 to 11, for the treatment, prevention, or management of RAGE-related disorders in patients requiring such treatment, prevention, or management.
  20. A pharmaceutical composition comprising a therapeutically effective amount of the fusion polypeptide described in claim 12, for treating, preventing, or managing RAGE-related disorders in patients requiring such treatment, prevention, or management.

Description

The present invention relates, in general terms, to a screening assay for identifying modulators of receptor activation associated with a particular disease and/or condition, such modulators, and a treatment method comprising the administration of such modulators. More specifically, the present invention relates to S100A 8/A9, RAG by RAGE ligands containing advanced glycation end products (AGEs) and HMGB1 Activated type 1 angiotensin receptor ( AT1R ), with or without regulation of E activation. and the ligand-independent mechanism of RAGE by specific coexisting activated G protein-coupled receptors (GPCRs), including activated CC chemokine receptor 2 (CCR2). Advanced glycation end product receptor (R) (also known as E-ligand-independent trans-activation) This invention relates to modulators of AGE activation. The invention also relates to a screening assay for identifying such modulators and a method for treating RAGE-related disorders using said modulators. Advanced glycation end product receptors (RAGEs) are polyvalent type I transmembrane glycoproteins belonging to the immunoglobulin (Ig) superfamily (Neeper et al., 1992). 5 Glycosylated RAGE proteins of 0–55 kDa are constitutively expressed in a limited range of cells (e.g., vascular endothelium, type II lung cells, leukocytes), but RAGE expression can be induced in most cell types and tissues after injury and inflammation (Ballinger et al., 2020). 05). RAGE expression is associated with cardiovascular disease (CVD), cancer, diabetes, and chronic kidney disease (CKD). It is significantly upregulated in important inflammatory and metabolic disorders, including but not limited to ischemic injury and Alzheimer's disease (Yan et al., 2010). Genetic deletion of the AGE gene, which codes for RAGE, is associated with several cancers (Malikh It has been previously demonstrated that RAGE provides protection from numerous diseases and disease processes in mice, including inflammatory diseases such as atherosclerosis and diabetic complications (Chuah et al., 2013). For example, in apolipoprotein E (apoE) knockout (KO) mice, RAGE deletion results in less plaque accumulation with age and reduced atherosclerosis accelerated by diabetes (Soro-Paavonen et al., 2008). Similarly, AGE Deletion of R can reduce renal impairment in diabetic mice without affecting glucose regulation (Thomas et al., 2005). AGER gene polymorphisms have been associated with numerous diseases and disease processes in humans, including but not limited to arthritis, atherosclerosis and diabetic complications, cancer risk, obesity, epilepsy, and cognitive impairment, including Alzheimer's disease. RAG by advanced glycation end products (AGEs) and non-AGE ligands (including members of the S100 cargranulin family of proteins, HMGB1, amyloid, and Mac-1) The binding of E to the extracellular domain activates a series of signaling cascades involved in inflammation, injury, and dysfunction, including nuclear factor kappa B (NFκB) and the renin-angiotensin-aldosterone system (RAAS). In experimental models, inhibition of ligand-mediated activation of RAGE using soluble decoy receptors attenuated atheroma formation and vascular damage (Schmidt et al. 1999). This means that the pathological effects of RAGE are partially mediated by ligand-mediated activation of RAGE in such a setting. The precise molecular mechanism by which RAGE is activated and all its biological effects are as follows: These clinically important signaling pathways are not fully understood, and therefore, their ability to target them remains unproven in clinical settings. The renin-angiotensin-aldosterone system (RAAS) is a crucial homeostatic pathway involved in the development and progression of many common diseases and disease processes. Angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor type 1 ( AT1R ) Renin-angiotensin-aldosterone system (RAA) blockers (inhibitors) Inhibition of RAAS is widely used in the management of many diseases and/or conditions, including hypertension, cardiovascular disease (CVD), heart failure, chronic kidney disease (CKD), and diabetic complications. RAAS inhibition is used in the prevention of diabetes (Tikellis et al., 2004) and neuroprotection (Theo). Shen Reineke et al., 2011), mitigation of the growth of certain cancers (Shen (et al., 2016) and gene deletion of AT1R , which gives mice a longer lifespan, has been shown to have advantages even in aging (Benigni et al., 2009). These effects of RAAS blockers are additive to and independent of the blood pressure reduction mediated by RAAS blockers. This is because equivalent blood pressure reduction by other drugs does not provide the same benefit (Lee et al., 1993). In particular, activation of AT1R by angiotensin II (Ang II) induces oxidative stress, activation of nuclear factor κB (NFκB), and inflammation via a pathway different from the one that causes vasoconstriction. Activation of the renin-angiotensin-aldosterone system (RAAS) i