Identification of a PCSK9-LDLR disruptor peptide with in vivo function

Brousseau, Margaret E.; Clairmont, Kevin B.; Spraggon, Glen; Flyer, Alec N.; Golosov, Andrei A.; Grosche, Philipp; Amin, Jakal; Andre, Jerome; Burdick, Debra; Caplan, Shari L.; Chen, Guanjing; Chopra, Raj; Ames, Lisa; Dubiel, Diana; Fan, Li; Gattlen, Raphael; Kelly-Sullivan, Dawn; Koch, Alexander W.; Lewis, Ian; Li, Jingzhou; Liu, Eugene; Lubicka, Danuta; Marzinzik, Andreas L.; Nakajima, Katsumasa; Nettleton, David O.; Ottl, Johannes; Pan, Meihui; Patel, Tajesh; Perry, Lauren M; Pickett, Stephanie; Poirier, Jennifer; Reid, Patrick; Pellé, Xavier; Seepersaud, Mohindra; Subramanian, Vanitha; Vera, Victoria; Xu, Mei; Yang, Lihua; Yang, Qing; Yu, Jinghua; Zhu, Guoming; Monovich, Lauren G.

doi:10.1016/j.chembiol.2021.08.012

Cited by 31 publications

(16 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a soluble protein and a ligand for LDLR. Extracellular PCSK9 binds to LDLR through protein-protein interactions and directly enters the lysosome as a PCSK9-LDLR complex for destruction, inhibiting LDLR recirculation and lowering plasma LDL-C levels [ 65 , 66 ]. The conversion of cholesterol into bile acid (BA) in the liver is its main metabolic pathway.…”

Section: Discussionmentioning

confidence: 99%

The Hypolipidemic Effect of Hawthorn Leaf Flavonoids through Modulating Lipid Metabolism and Gut Microbiota in Hyperlipidemic Rats

Weng

Cui

et al. 2022

Evidence-Based Complementary and Alternative Medicine

View full text Add to dashboard Cite

Objective. The purpose of this study was to explore the potential mechanisms of the lipid-regulating effects and the effect on modulating the gut microbiota of hawthorn leaf flavonoids (HLF) in the high-fat diet-induced hyperlipidemic rats. Methods. The hypolipidemic effect of HLF was investigated in the high-fat diet-induced hyperlipidemic rats. The action targets of HLF in the treatment of hyperlipidemia were predicted by network pharmacology and KEGG enrichment bubble diagram, which were verified by the test of western blotting. Meanwhile, we used 16S rRNA sequencing to evaluate the effects of HLF on the microbes. Results. The results of animal experiments showed that HLF could reduce the body weight and regulate the levels of serum lipid in high-fat diet (HFD) rats. Meanwhile, for the related targets of cholesterol metabolism, HLF could significantly upregulate the expression of LDLR, NR1H3, and ABCG5/ABCG8; reduce the expression of PCSK9; and increase the level of CYP7A1 in the intestinal tissue, whereas cholesterol biosynthetic protein expressions including HMGCR and SCAP were lowered by HLF. In addition, HLF increased the activities of plasma SOD, CAT, and GSH-Px and decreased the levels of Casp 1, NLRP3, IL-1β, IL-18, and TNF-α, improving the degree of hepatocyte steatosis and inflammatory infiltration of rats. Notably, HLF significantly regulated the relative abundance of major bacteria such as g_Lactobacillus, g_Anaerostipes, g_[Eubacterium]_hallii_group, g_Fusicatenibacter, g_Akkermansia, and g_Collinsella. Synchronously, we found that HLF could regulate the disorder of plasma HEPC and TFR levels caused by HFD. Conclusion. This study demonstrates that HLF can regulate metabolic hyperlipidemia syndromes and modulate the relative abundance of major bacteria, which illustrated that it might be associated with the modulation of gut microbiota composition and metabolites.

show abstract

Section: Discussionmentioning

confidence: 99%

The Hypolipidemic Effect of Hawthorn Leaf Flavonoids through Modulating Lipid Metabolism and Gut Microbiota in Hyperlipidemic Rats

Weng

Cui

et al. 2022

Evidence-Based Complementary and Alternative Medicine

View full text Add to dashboard Cite

show abstract

“…The recycling of LDLR induced by PCSK9 is mainly achieved through the binding between the epidermal growth factor-like repeat-A (EGF-A) domain on LDLR and the catabolic domain on PCSK9, for which PCSK9 inhibitors have been developed to bind with EGF-A site on PCSK9, and therefore silence its functions, prevent degradations of LDLR, and therefore reduce LDL-C levels [ 27 ]. In recent years, several peptide inhibitors for PCSK9 have been found or developed, including both analogs of EGF-A and non-EGF-A-based peptides identified from randomized peptide sequences, which are proven to effectively bind with the EGF-A site on PCSK9 and inhibit the PCSK9:LDLR interface with high potency [ 27 , 28 ]. More recently, PCSK9 has been found to be actively involved in host defense in infectious diseases (e.g., sepsis, hepatitis), so there could be possible interactions between PCSK9 and various immunomodulators, including antimicrobial peptide LL-37 [ 29 , 30 ] Future studies are warranted to investigate whether LL-37 could bind with PCSK9 to restore the function of LDLR and its impacts on long-term LDL-C levels, which would provide essential evidence about whether LL-37 could be a potential antagonist for PCSK9.…”

Section: Discussionmentioning

confidence: 99%

Prognostic Impacts of LL-37 in Relation to Lipid Profiles of Patients with Myocardial Infarction: A Prospective Cohort Study

et al. 2022

View full text Add to dashboard Cite

Background. In vivo studies show that LL-37 inhibits the progression of atherosclerosis and predicts a lower risk of recurrent ischemia in patients with acute myocardial infarction (AMI), which could be mediated by the modulation of lipid metabolism. The current study aimed to investigate the effects of various lipid contents on the prognostic impacts of LL-37 in patients with AMI. Methods. A total of 1567 consecutive AMI patients were prospectively recruited from March 2017 to January 2020. Patients were firstly stratified into two groups by the median level of LL-37 and then stratified by levels of various lipid contents and proprotein convertase subtilisin/kexin type 9 (PCSK9). Cox regression with multiple adjustments was performed to analyze associations between LL-37, lipid profiles, PCSK9, and various outcomes. The primary outcome was major adverse cardiovascular event (MACE), a composite of all-cause death, recurrent MI, and ischemic stroke. Results. During a median follow-up of 786 (726–1107) days, a total of 252 MACEs occurred. A high level of LL-37 was associated with lower risk of MACE in patients with elevated lipoprotein(a) (≥300 mg/L, hazard ratio (HR): 0.49, 95% confidence interval (CI): 0.29–0.86, p = 0.012) or PCSK9 levels above the median (≥47.4 ng/mL, HR: 0.57, 95% CI: 0.39–0.82, p < 0.001), which was not observed for those without elevated lp(a) (<300 mg/L, HR: 0.96, 95% CI: 0.70–1.31, p = 0.781, p interaction = 0.035) or PCSK9 (<47.4 ng/mL, HR: 1.02, 95% CI: 0.68–1.54, p = 0.905, p interaction = 0.032). Conclusions. For patients with AMI, a high level of LL-37 was associated with lower ischemic risk among patients with elevated lp(a) and PCSK9.

show abstract

“…To the best of our knowledge, of the 20 published studies, only three have progressed beyond in vitro activity optimization to show in vivo pharmacokinetic characterization. 24,25,27 Albumin-binding through lipidation has emerged as an industry validated half-life extension strategy that has successfully generated five approved drugs in the last two decades 31−35 but has not been applied to PCSK9 inhibitory peptides. Human serum albumin (HSA) is the most abundant protein in blood with a long half-life of ∼19 days.…”

Section: ■ Introductionmentioning

confidence: 99%

“…To retain this selectivity but improve affinity, we developed a design approach utilizing phage display to engineer a cyclization linker onto Pep2–8, resulting in an analogue called P9–38 (Figure C), which has ∼100-fold improved PCSK9 binding affinity compared to Pep2–8 and activity in the nanomolar range . Additional peptides displaying high in vitro activity have been recently reported to be of therapeutic potential, − but a broader understanding of their in vivo efficacy is required.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Improving Stability Enhances In Vivo Efficacy of a PCSK9 Inhibitory Peptide

et al. 2022

View full text Add to dashboard Cite

Optimization of peptide stability is essential for the development of peptides as bona fide alternatives to approved monoclonal antibodies. This is clearly the case for the many peptides reported to antagonize proprotein convertase subtilisin-like/kexin type 9 (PCSK9), a clinically validated target for lowering cholesterol. However, the effects of optimization of stability on in vivo activity and particularly the effects of binding to albumin, an emerging drug design paradigm, have not been studied for such peptide leads. In this study, we optimized a PCSK9 inhibitory peptide by mutagenesis and then by conjugation to a short lipidated tag to design P9-alb fusion peptides that have strong affinity to human serum albumin. Although attachment of the tag reduced activity against PCSK9, which was more evident in surface plasmon resonance binding and enzyme-linked immunosorbent competition assays than in cellular assays of activity, activity remained in the nanomolar range (∼40 nM). P9-alb peptides were exceptionally stable in human serum and had half-lives exceeding 48 h, correlating with longer half-lives in mice (40.8 min) compared to the unconjugated peptide. Furthermore, the decrease in in vitro binding was not deleterious to in vivo function, showing that engendering albumin binding improved low-density lipoprotein receptor recovery and cholesterol-lowering activity. Indeed, the peptide P9-albN2 achieved similar functional endpoints as the approved anti-PCSK9 antibody evolocumab, albeit at higher doses. Our study illustrates that optimization of stability instead of binding affinity is an effective way to improve in vivo function.

show abstract

Identification of a PCSK9-LDLR disruptor peptide with in vivo function

Cited by 31 publications

References 45 publications

The Hypolipidemic Effect of Hawthorn Leaf Flavonoids through Modulating Lipid Metabolism and Gut Microbiota in Hyperlipidemic Rats

The Hypolipidemic Effect of Hawthorn Leaf Flavonoids through Modulating Lipid Metabolism and Gut Microbiota in Hyperlipidemic Rats

Prognostic Impacts of LL-37 in Relation to Lipid Profiles of Patients with Myocardial Infarction: A Prospective Cohort Study

Improving Stability Enhances In Vivo Efficacy of a PCSK9 Inhibitory Peptide

Contact Info

Product

Resources

About