bims-cepepe 2025-05-25 papers

bims-cepepe

Biomed News

on Cell-penetrating peptides

Issue of 2025–05–25
sixteen papers selected by
Henry Lamb, Queensland University of Technology

RaPID discovery of cell-permeable helical peptide inhibitors con-taining cyclic β-amino acids against SARS-CoV-2 main protease.
Cyclic peptide structure prediction and design using AlphaFold2.
Identification of Potent Cell Penetrating, Nontoxic Peptides for Inhibiting MDM2-p53 Interactions: Characterization of Anticancer Peptides via Molecular Dynamics Simulations.
Lactate dehydrogenase A-coupled NAD+ regeneration is critical for acute myeloid leukemia cell survival.
HighPlay: Cyclic Peptide Sequence Design Based on Reinforcement Learning and Protein Structure Prediction.
A Cyclized Helix-Loop-Helix Peptide as a Molecular Scaffold for Cell-Membrane Permeable Inhibitors for the Interaction between Estrogen Receptor α and Coactivator SRC1.
An mRNA-display derived cyclic peptide scaffold reveals the substrate binding interactions of an N-terminal cysteine oxidase.
Increased Throughput of Combined Stability Testing and Metabolite Identification Using a Sample Multiplexing Strategy for the Optimization of Engineered Cyclic Peptide Drugs.
A super versatile flexizyme system with phenol esters for genetic code reprogramming.
Enhanced delivery of camptothecin to colorectal carcinoma using a tumor-penetrating peptide targeting p32.
Research progress on the role of Claudin family proteins in mediating blood-brain barrier selective permeability in tumor metastasis.
[68Ga]Ga-NOTA-C-IPB-AIT: A novel radiotracer for in vivo detection of TREM-1 expression.
Reengineering of a Proteomimetic Pan-Ras Inhibitor into a Ras Degrader.
Can pretreatment lactate dehydrogenase to albumin ratio predict pathological complete response after neoadjuvant chemotherapy in breast cancer patients?
Lipopeptide-mediated delivery of CRISPR/Cas9 ribonucleoprotein complexes for gene editing and correction.
Meeting Report: DMDG Peptide and Oligonucleotide ADME Workshop 2024.

RSC Chem Biol. 2025 May 21.

RaPID discovery of cell-permeable helical peptide inhibitors con-taining cyclic β-amino acids against SARS-CoV-2 main protease.

Marina Kawai, Tika R Malla, H T Henry Chan, Anthony Tumber, Lennart Brewitz, Eidarus Salah, Naohiro Terasaka, Takayuki Katoh, Akane Kawamura, Christopher J Schofield, Fernanda Duarte, Hiroaki Suga.

Structurally constrained cyclic β-amino acids are attractive building blocks for peptide drugs because they induce unique and stable conformations. Introduction of (1S,2S)-2-aminocyclopentanecarboxylic acid [(1S,2S)-2-ACPC] into peptides stabilizes helical conformations, so improving proteolytic stability and cell membrane permeability. We report on the ribosomal synthesis of a helical peptide library incorporating (1S,2S)-2-ACPC at every third position and its application for the discovery of SARS-CoV-2 main protease (Mpro) inhibitors. We identified two peptide sequences containing multiple (1S,2S)-2-ACPC residues, which exhibit helical conformations and superior proteolytic stability compared with their α-Ala or β-Ala counterparts. Studies using the chloroalkane cell-penetration assay showed that their cell permeability values (CP50) are comparable with or even slightly better than that of the cell-penetrating nona-arginine (R9) peptide. The new approach is thus a highly efficient method that combines a helical peptide library containing structurally constrained cyclic β-amino acids with the classical RaPID discovery method, enabling de novo discovery of proteolytically stable and cell-penetrating bioactive peptides that target intracellular proteins.

DOI: https://doi.org/10.1039/d5cb00021a
Nat Commun. 2025 May 21. 16(1): 4730

Cyclic peptide structure prediction and design using AlphaFold2.

Stephen A Rettie, Katelyn V Campbell, Asim K Bera, Alex Kang, Simon Kozlov, Yensi Flores Bueso, Joshmyn De La Cruz, Maggie Ahlrichs, Suna Cheng, Stacey R Gerben, Mila Lamb, Analisa Murray, Victor Adebomi, Guangfeng Zhou, Frank DiMaio, Sergey Ovchinnikov, Gaurav Bhardwaj.

Small cyclic peptides have gained significant traction as a therapeutic modality; however, the development of deep learning methods for accurately designing such peptides has been slow, mostly due to the lack of sufficiently large training sets. Here, we introduce AfCycDesign, a deep learning approach for accurate structure prediction, sequence redesign, and de novo hallucination of cyclic peptides. Using AfCycDesign, we identified over 10,000 structurally-diverse designs predicted to fold into the designed structures with high confidence. X-ray crystal structures for eight tested de novo designed sequences match very closely with the design models (RMSD < 1.0 Å), highlighting the atomic level accuracy in our approach. Further, we used the set of hallucinated peptides as starting scaffolds to design binders with nanomolar IC50 against MDM2 and Keap1. The computational methods and scaffolds developed here provide the basis for the custom design of peptides for diverse protein targets and therapeutic applications.

DOI: https://doi.org/10.1038/s41467-025-59940-7
J Pept Sci. 2025 Jul;31(7): e70033

Identification of Potent Cell Penetrating, Nontoxic Peptides for Inhibiting MDM2-p53 Interactions: Characterization of Anticancer Peptides via Molecular Dynamics Simulations.

Pradeep Pant, Aman Sehgal, Tushar Gupta, Pradeep Sharma.

  Inhibiting MDM2-p53 interactions is a crucial therapeutic strategy in cancer treatment, as it can restore the tumor suppressor activity of p53 and inhibit tumor progression. Peptide inhibitors have shown promise in targeting this interaction; however, their optimization for in vivo use often encounters challenges, particularly in cellular uptake. The present study addresses this limitation by identifying cell-penetrating peptides (CPPs) that are predicted to be nontoxic to humans and capable of inhibiting the MDM2-p53 interaction. We utilized a comprehensive CPP database to extract unmodified peptides, focusing on those predicted to be nontoxic. Selected candidates were subjected to molecular docking followed by 500-ns all-atom explicit-solvent molecular dynamics (MD) simulations, performed in triplicates, to evaluate their binding stability and affinity with MDM2. Binding affinity calculations using MM-PBSA, AREA AFFINITY, and PRODIGY revealed that two peptides consistently exhibited stable binding to MDM2 and demonstrated higher affinity compared with the p53 reference fragment. These peptides not only maintained favorable interactions throughout the simulations but also showed strong potential to disrupt MDM2-p53 binding and reactivate p53 function. The findings highlight these peptides as promising nontoxic anticancer agents and provide a strong foundation for the development of peptide-based therapeutics targeting the MDM2-p53 interaction.

Keywords:  MDM2; anticancer peptides; cell penetrating peptides; molecular dynamics simulations; p53

DOI:  https://doi.org/10.1002/psc.70033
Cancer Metab. 2025 May 19. 13(1): 22

Lactate dehydrogenase A-coupled NAD+ regeneration is critical for acute myeloid leukemia cell survival.

Ayşegül Erdem, Séléna Kaye, Francesco Caligiore, Manuel Johanns, Fleur Leguay, Jan Jacob Schuringa, Keisuke Ito, Guido Bommer, Nick van Gastel.

   BACKGROUND: Enhanced glycolysis plays a pivotal role in fueling the aberrant proliferation, survival and therapy resistance of acute myeloid leukemia (AML) cells. Here, we aimed to elucidate the extent of glycolysis dependence in AML by focusing on the role of lactate dehydrogenase A (LDHA), a key glycolytic enzyme converting pyruvate to lactate coupled with the recycling of NAD+.
METHODS: We compared the glycolytic activity of primary AML patient samples to protein levels of metabolic enzymes involved in central carbon metabolism including glycolysis, glutaminolysis and the tricarboxylic acid cycle. To evaluate the therapeutic potential of targeting glycolysis in AML, we treated AML primary patient samples and cell lines with pharmacological inhibitors of LDHA and monitored cell viability. Glycolytic activity and mitochondrial oxygen consumption were analyzed in AML patient samples and cell lines post-LDHA inhibition. Perturbations in global metabolite levels and redox balance upon LDHA inhibition in AML cells were determined by mass spectrometry, and ROS levels were measured by flow cytometry.
RESULTS: Among metabolic enzymes, we found that LDHA protein levels had the strongest positive correlation with glycolysis in AML patient cells. Blocking LDHA activity resulted in a strong growth inhibition and cell death induction in AML cell lines and primary patient samples, while healthy hematopoietic stem and progenitor cells remained unaffected. Investigation of the underlying mechanisms showed that LDHA inhibition reduces glycolytic activity, lowers levels of glycolytic intermediates, decreases the cellular NAD+ pool, boosts OXPHOS activity and increases ROS levels. This increase in ROS levels was however not linked to the observed AML cell death. Instead, we found that LDHA is essential to maintain a correct NAD+/NADH ratio in AML cells. Continuous intracellular NAD+ supplementation via overexpression of water-forming NADH oxidase from Lactobacillus brevis in AML cells effectively increased viable cell counts and prevented cell death upon LDHA inhibition.
CONCLUSIONS: Collectively, our results demonstrate that AML cells critically depend on LDHA to maintain an adequate NAD+/NADH balance in support of their abnormal glycolytic activity and biosynthetic demands, which cannot be compensated for by other cellular NAD+ recycling systems. These findings also highlight LDHA inhibition as a promising metabolic strategy to eradicate leukemic cells.

Keywords:  Acute myeloid leukemia; Cancer metabolism; Glycolysis; Lactate dehydrogenase A; NAD+ ; Redox balance

DOI:  https://doi.org/10.1186/s40170-025-00392-4
J Med Chem. 2025 May 23.

HighPlay: Cyclic Peptide Sequence Design Based on Reinforcement Learning and Protein Structure Prediction.

Huitian Lin, Cheng Zhu, Tianfeng Shang, Ning Zhu, Kang Lin, Chengyun Zhang, Xiang Shao, Xudong Wang, Hongliang Duan.

The structural diversity and good biocompatibility of cyclic peptides have led to their emergence as potential therapeutic agents. Existing cyclic peptide design methods, whether traditional or emerging AI-assisted, rely on a multitude of experiments and face challenges such as limited molecular diversity, high cost, and time-consuming. In this study, we propose HighPlay, which integrates reinforcement learning (MCTS) with the HighFold structural prediction model to design cyclic peptide sequences based solely on the target protein sequence information, to achieve the synergistic optimization of cyclic peptide sequences and binding sites and to dynamically explore the sequence space without the need for predefined target information. The model was applied to the design of cyclic peptide sequences for three different targets, which were screened and verified through molecular dynamics simulations, demonstrating good binding affinity. Specifically, the cyclic peptide sequences designed for the TEAD4 target exhibited micromolar-level affinity in further experimental validation.

DOI: https://doi.org/10.1021/acs.jmedchem.5c00896
Chembiochem. 2025 May 23. e202500232

A Cyclized Helix-Loop-Helix Peptide as a Molecular Scaffold for Cell-Membrane Permeable Inhibitors for the Interaction between Estrogen Receptor α and Coactivator SRC1.

Daisuke Fujiwara, Shunsuke Inaura, Yuna Tanaka, Sayoko Ito-Harashima, Aasako Yamaguchi-Nomoto, Masanobu Kawanishi, Takashi Yagi, Ikuhiko Nakase, Ikuo Fujii.

  The molecular design of inhibitors against intracellular protein-protein interactions (PPIs) is of interest for drug discovery and chemical biology. Here, we designed a novel cyclized helix-loop-helix (cHLH) peptide that inhibited the intracellular PPI between estrogen receptor alpha (ERα) and coactivator SRC1. The peptide, cHLH-ERα, bound to ERα and inhibited the interaction between ERα and the coactivator SRC1. Cellular imaging and yeast reporter assays showed that cHLH-ERα penetrated the cell membrane and exhibited antagonistic activity against ERα-SRC1 to inhibit the growth of a breast cancer cell.

Keywords:  Cell membrane permeability; Nuclear Receptor; Protein-protein interaction; inhibitor; peptide

DOI:  https://doi.org/10.1002/cbic.202500232
Nat Commun. 2025 May 22. 16(1): 4761

An mRNA-display derived cyclic peptide scaffold reveals the substrate binding interactions of an N-terminal cysteine oxidase.

Yannasittha Jiramongkol, Karishma Patel, Jason Johansen-Leete, Joshua W C Maxwell, Yiqun Chang, Jonathan J Du, Toby Passioura, Kristina M Cook, Richard J Payne, Mark D White.

N-terminal cysteine oxidases (NCOs) act as enzymatic oxygen (O2) sensors, coordinating cellular changes to hypoxia in animals and plants. They regulate the O2-dependent stability of proteins bearing an N-terminal cysteine residue through the N-degron pathway. Despite their important role in hypoxic adaptation, which renders them potential therapeutic and agrichemical targets, structural information on NCO substrate binding remains elusive. To overcome this challenge, we employed a unique strategy by which a cyclic peptide inhibitor of the mammalian NCO, 2-aminoethanethiol dioxygenase (ADO), was identified by mRNA display and used as a scaffold to graft substrate moieties. This allowed the determination of two substrate analogue-bound crystal structures of ADO. Key binding interactions were revealed, including bidentate coordination of the N-terminal residue at the metal cofactor. Subsequent structure guided mutagenesis identified aspartate-206 as an essential catalytic residue, playing a role in reactive oxygen intermediate orientation or stabilisation. These findings provide fundamental information on ADO substrate interactions, which can elucidate enzyme mechanism and act as a platform for chemical discovery.

DOI: https://doi.org/10.1038/s41467-025-59960-3
J Am Soc Mass Spectrom. 2025 May 23.

Increased Throughput of Combined Stability Testing and Metabolite Identification Using a Sample Multiplexing Strategy for the Optimization of Engineered Cyclic Peptide Drugs.

Congliang Sun, Mark T Cancilla, Bahanu Habulihaz, Lisa A Vasicek, Daniel S Spellman, Sven Hackbusch, Sebastien Morin.

Engineered cyclic peptides (ECPs) have been in the spotlight as novel drug modalities for challenging therapeutic targets. Oral delivery of engineered cyclic peptides benefits from ease of administration. However, one of the main hurdles in developing orally effective peptide drugs is their potential metabolic instability due to enzymatic degradation. To that end, in vitro experiments with simulated intestinal fluid (SIF) have been used to assess drug metabolic stability in the gastrointestinal tract. Currently, metabolic stability evaluations and biotransformation assessments are performed separately, which can be time-consuming and result in complex data analysis. Presented here is a sample multiplexing strategy to address these challenges by leveraging a Thermo Scientific Orbitrap Astral mass spectrometer with two complementary analyzers, enabling the simultaneous analysis of metabolic stability from the Orbitrap full scan and biotransformation from the Astral analyzer from one sample injection. Furthermore, we demonstrate that 10 engineered cyclic peptides can be pooled into one sample injection without compromising the data quality to decrease the instrument run time and improve the throughput of the assay.

DOI: https://doi.org/10.1021/jasms.5c00113
Nat Commun. 2025 May 20. 16(1): 4671

A super versatile flexizyme system with phenol esters for genetic code reprogramming.

Yun-Kyu Choi, Takayuki Katoh, Adam Beattie, Hiroaki Suga.

Genetic code reprogramming enables ribosomal incorporation of multiple nonproteinogenic amino acids (npAAs), supporting bioactive peptide development. Flexizyme technology with npAA-benzyl(thio)esters (BZEs) as acyl-donors has been crucial for preparing diverse npAA-tRNAs. However, low acylation yields for some npAAs hinder peptide library construction. Here we report a versatile flexizyme system using phenol esters as alternative acyl-donors. Computational pKa predictions guided the synthesis of five phenol esters, which are mild enough to prevent random aminoacylation yet reactive with tRNA 3'-hydroxy groups. Among them, 3-nitrophenol (3NP) was chosen to prepare 18 structurally distinct npAA-3NPs, demonstrating direct tRNAs aminoacylation in 15-41% yields. Moreover, the flexizyme eFx drastically enhances aminoacylation efficiency, yielding near-quantitative conversion for some npAAs, e.g. cyclic β-npAAs. This eFx/npAA-3NP system facilitates access to diverse npAA-tRNAs, expanding ribosomal synthesis of nonstandard peptides, including macrocyclic structures. Thus, our approach provides an efficient tool for constructing peptide libraries with multiple npAA building blocks.

DOI: https://doi.org/10.1038/s41467-025-59921-w
Acta Biomater. 2025 May 14. pii: S1742-7061(25)00361-7. [Epub ahead of print]

Enhanced delivery of camptothecin to colorectal carcinoma using a tumor-penetrating peptide targeting p32.

Yanhao Jiang, Zhiren Wang, Wenpan Li, Teng Ma, Mengwen Li, Shuang Wu, Ethan Lin, Karlie Elizabeth Flader, Mengjiao Ma, Mengyang Chang, Hongmin Li, Wei Wang, Jianqin Lu.

  Camptothesome, a sphingomyelin (SM)-conjugated camptothecin (CPT) vesicular nanotherapeutic, addresses the poor solubility and lactone instability of CPT while enhancing drug loading, pharmacokinetics, and tumor distribution compared to CPT physically entrapped in conventional liposomes. Despite these improvements, the tumor uptake remains limited. To further enhance the tumor delivery efficiency and minimize the off-target distribution, we functionalize Camptothesome with the LinTT1 peptide, a CendR motif, which binds to overexpressed p32 proteins on tumor cell surface, initiating effective transcytosis for deep tumor penetration. Via systematic screening, the optimal peptide ratio on Camptothesome is identified. LinTT1/Camptothesome significantly increases cancer cell uptake without affecting normal cell internalization, resulting in enhanced anti-colorectal cancer cells activity. Additionally, decorating Camptothesome with the LinTT1 cell-penetrating peptide enables effective transcytosis via a Golgi-dependent intracellular trafficking mechanism, significantly improving the intratumoral delivery while reducing distribution to normal tissues. In a human HCT116 xenograft colorectal cancer (CRC) mouse model, LinTT1/Camptothesome demonstrates superior antitumor efficacy compared to both Camptothesome and Onivyde by upregulating cleaved caspase-3 and γH2AX. Our study substantiates the potential of leveraging a tumor-penetrating peptide to enhance the tumor delivery efficiency of Camptothesome, maximizing its therapeutic index for improved treatment of human CRC. STATEMENT OF SIGNIFICANCE: Despite the improved tumor delivery achieved by Camptothesome, its tumor distribution and penetration remain limited. This is because the enhanced permeability and retention effect only facilitates nanotherapeutic distribution to tumor periphery through leaky vasculature. The C-end Rule (CendR) motif-neuropilin receptor system enhances tumor-homing peptides by binding to cellular surface receptors, triggering transcytosis. Herein, LinTT1, the most potent CendR peptide that binds to the overexpressed p32 receptor on cancer cells, was effectively engineered onto Camptothesome using thiol-maleimide lipid chemistry. The LinTT1/Camptothesome significantly enhanced tumor uptake and penetration while minimizing accumulation in normal tissues, demonstrating remarkable anticancer efficacy in a human xenograft colorectal cancer model. Our findings highlight the critical role of tumor-homing peptides in unlocking the full therapeutic potential of Camptothesome.

Keywords:  Camptothesome; Colorectal cancer; Nanotherapeutics; Tumor-homing LinTT1 peptide; p32

DOI:  https://doi.org/10.1016/j.actbio.2025.05.036
Am J Transl Res. 2025 ;17(4): 2411-2421

Research progress on the role of Claudin family proteins in mediating blood-brain barrier selective permeability in tumor metastasis.

Dianfang Wei, Ming Xu, Weitong Cui, Xiaoyu Feng, Lei Wei, Kai Wang, Shuai Li, Shuping Gao.

  The blood-brain barrier (BBB) serves as a critical protective mechanism for the central nervous system (CNS), controlling the selective passage of molecules between the brain and the bloodstream. Claudin proteins, key components of tight junctions, play a central role in maintaining BBB integrity and regulating its permeability. Recent research has increasingly focused on how Claudins contribute to brain metastasis, where tumor cells alter Claudin expression to breach the BBB and invade brain tissue. While Claudin family such as Claudin-1 and Claudin-5 are essential for maintaining BBB function, their dysregulation in tumor cells facilitates BBB disruption, promoting metastasis. This review explores the dual role of Claudins in tumor progression, detailing how they regulate BBB permeability and enable tumor cells to cross the barrier. Additionally, we discuss the potential of Claudin proteins as therapeutic targets in cancer treatment, offering new insights into mechanisms of brain metastasis.

Keywords:  BBB; Claudin; cancer treatment; selective permeability; tumor metastasis

DOI:  https://doi.org/10.62347/GGGX3909
Bioorg Chem. 2025 May 19. pii: S0045-2068(25)00491-2. [Epub ahead of print]162 108611

[68Ga]Ga-NOTA-C-IPB-AIT: A novel radiotracer for in vivo detection of TREM-1 expression.

Yuan Pan, Xiaoli Zhang, Can Diao, Jinglin Zhang, Yixiang Lin, Xiangyang Sun, Min Li, Feng Gao.

  Triggering receptor expressed on myeloid cells 1 (TREM-1) plays a critical role in the initiation and advancement of cancer. The noninvasive and rapid detection of TREM-1 expression provides a significant value for oncologic diagnosis, tumor evaluation and the guide of TREM-1-targeted therapy. In this study, we developed three peptide-based TREM-1 targeted radiotracers and evaluated their radiochemical and biological properties, in order to identify the candidate for the detection of TREM-1 expression on tumors. Among three radiotracers, [68Ga]Ga-NOTA-C-IPB-AIT showed a high radiochemical purity (> 98 %), excellent stability and hydrophilicity, favorable binding affinity, remarkable tumor uptake, ideal tumor-to-muscle ratio, and prolonged tumor retention, indicating that it is promising for potential use in TREM-1 expression detection.

Keywords:  (68)Ga; PET; Radiolabel; Radiotracer

DOI:  https://doi.org/10.1016/j.bioorg.2025.108611
Angew Chem Int Ed Engl. 2025 May 19. e202507092

Reengineering of a Proteomimetic Pan-Ras Inhibitor into a Ras Degrader.

Joseph F Ongkingco, Seong Ho Hong, Eugene D Toth, Thu Nguyen, Paramjit S Arora.

  Bifunctional ligands that can coax protein-protein interactions have become attractive therapeutic modalities. Herein, we describe conformationally defined helix dimers as proteomimetic molecular glues. The helix dimers can be rationally designed toengage helical protein interfaces. We previously described a synthetic Sos protein mimic, CHDSOS, as a Ras ligand that inhibits wild-type and oncogenic Ras signaling. This Sos proteomimetic consisted of a crosslinked helix dimer (CHD) that reproduced two helical domains, termed aH and aI, from Sos. The native aH helix of Sos constitutes the primary contact surface for Sos while aI has minimal engagement. We conjectured that the aI domain of CHDSOS could be reengineered to preserve Ras binding while engaging another protein to fully leverage the contact residues available in a proteomimetic. Herein, we incorporate a second distinct binding epitope into CHDSOS, thereby generating a bispecific proteomimetic. This secondary epitope was designed based on the p53 activation domain to engage the E3 ligase MDM2 and induce complexation with Ras. The resulting lead proteomimetic, CHDBI4, associates with both MDM2 and Ras and demonstrates reduction of cellular Ras levels. Overall, the study offers a proof-of-concept for the development of a bispecific proteomimetic scaffold to target multiple protein interfaces.

Keywords:  Molecular glues; Protein-protein interactions; Ras; constrained peptides; proteomimetics

DOI:  https://doi.org/10.1002/anie.202507092
J Med Biochem. 2025 Mar 21. 44(2): 339-346

Can pretreatment lactate dehydrogenase to albumin ratio predict pathological complete response after neoadjuvant chemotherapy in breast cancer patients?

Gözde Savaş, Nazan Günel, Ahmet Özet.

   Background: This study aims to evaluate the predictive significance of platelet lymphocyte ratio (PLR), neutrophillymphocyte ratio (NLR), lymphocyte monocyte ratio (LMR), systemic immune-inflammation (SII), prognostic nutritional index (PNI), haemoglobin, albumin, lymphocyte, and platelet (HALP) score and lactate dehydrogenase to albumin ratio (LAR) for pCR in breast cancer with neoadjuvant chemotherapy (NACT).
Methods: A total of 121 patients who received NACT between February 2012 and November 2021 were included. LAR, NLR, PLR, MLR, SII, PNI and HALP were calculated using formulas. The cut-off value for markers was obtained by Receiver operating characteristic curve (ROC) analyses. Independent predictive factors for pCR were determined using multivariate regression analysis.
Results: The pCR rate was achieved in 31.4% of patients. Median values of NLR, PLR, MLR, SII, PNI and HALP were similar in pCR (+) and pCR (-) (p>0.05). The median LAR value was significantly higher in pCR (+) than in pCR (-) (50.80 vs 42.62, respectively (p=0.002)). The optimal cut-off value of LAR was 46.27. Multivariate analysis showed that LAR ≥46.27 and HER-2 positivity were the independent predictive factors for pCR [OR=2.851 (95% CI=1.142-7.119, P=0.025), OR=3.431 (95% CI= 0.163-10.123, P=0.026), respectively].
Conclusions: LAR is a simple, inexpensive, and convenient method for predicting pCR in breast cancer with NACT.

Keywords:  LAR; LDH; albumin; breast cancer; complete response; neoadjuvant chemotherapy

DOI:  https://doi.org/10.5937/jomb0-43900
J Control Release. 2025 May 17. pii: S0168-3659(25)00474-2. [Epub ahead of print]383 113854

Lipopeptide-mediated delivery of CRISPR/Cas9 ribonucleoprotein complexes for gene editing and correction.

Mert Öktem, Thai Hoang Nguyen, Esmeralda D C Bosman, Marcel H A M Fens, Massimiliano Caiazzo, Enrico Mastrobattista, Zhiyong Lei, Olivier G de Jong.

  CRISPR/Cas gene editing is a highly promising technology for the treatment and even potential cure of genetic diseases. One of the major challenges for its therapeutic use is finding safe and effective vehicles for intracellular delivery of the CRISPR/Cas9 ribonucleoprotein (RNP) complex. In this study, we tested and characterized a series of novel fatty acid-modified versions of a previously reported Cas9 RNP carrier, consisting of a complex of the cell-penetrating peptide (CPP) LAH5 with Cas9 RNP and homology-directed DNA repair templates. Comparative experiments demonstrated that RNP/peptide nanocomplexes showed various improvements depending on the type of fatty acid modification. These improvements included enhanced stability in serum, improved membrane disruption capability and increased transfection efficacy. Cas9 RNP/oleic acid LAH5 peptide nanocomplexes showed the overall best performance for both gene editing and correction. Moreover, Cas9 RNP/oleic acid LAH5 nanocomplexes significantly protected the Cas9 protein cargo from enzymatic protease digestion. In addition, in vivo testing demonstrated successful gene editing after intramuscular administration. Despite the inherent barriers of the tightly organized muscle tissues, we achieved approximately 10 % gene editing in the skeletal muscle tissues when targeting the CAG-tdTomato locus in the transgenic Ai9 Cre-LoxP reporter mouse strain and 7 % gene editing when targeting the Ccr5 gene, without any observable short-term toxicity. In conclusion, the oleic acid-modified LAH5 peptide is an effective delivery platform for direct Cas9/RNP delivery, and holds great potential for the development of new CRISPR/Cas9-based therapeutic applications for the treatment of genetic diseases.

Keywords:  CRISPR-Cas9; Cell-penetrating peptides; Fatty-acid modifications; Gene editing; Intramuscular delivery

DOI:  https://doi.org/10.1016/j.jconrel.2025.113854
Xenobiotica. 2025 May 20. 1-10

Meeting Report: DMDG Peptide and Oligonucleotide ADME Workshop 2024.

Inga Bjørnsdottir, Ralf Lotz, Bo Lindmark, Steve Hood, Jesper Kammersgaard Christensen.

  Challenges within peptide and oligonucleotide ADME (absorption, distribution, metabolism and elimination) and scientific ideas on how to solve them were presented and discussed at the DMDG (Drug Metabolism and Discussion Group) Peptide and Oligonucleotide ADME Workshop 2024 (1st September 2024). This meeting report summarises the presentations and discussions from this workshop.The following topics were covered:Metabolism of conjugated peptides & proteinsStrategies for increased metabolic stability of peptidesIQ's take on metabolism & excretion studies for peptidesDMPK challenges related to oligonucleotides during developmentImaging of oligonucleotidesIdentification of extrahepatic targets of oligonucleotidesIn vitro systems for metabolism studies of oligonucleotidesNovel hybridization LC-MS/MS approach for quantitative analysis of oligonucleotidesPKPD, PBPK & modelling.

Keywords:  PKPD and PBPK modelling and hybrid LC-MS/MS; drug-drug interactions; extrahepatic delivery of siRNAs; human unique metabolite; in vitro assays; oligonucleotides; peptides

DOI:  https://doi.org/10.1080/00498254.2025.2506702