Jean-Michel Brunel

UMR-MD1 "Membranes et Cibles Thérapeutiques"

Doxycycline, a semi‐synthetic tetracycline, is a widely used antibiotic for treating mild‐to‐moderate infections, including skin problems. However, its anti‐inflammatory and antioxidant properties, combined with its ability to interfere with α‐synuclein aggregation, make it an attractive candidate for repositioning in Parkinson's disease. Neverthel...

The Streptomyces strain G222, isolated from a Vietnamese marine sediment, was confidently identified by 16S rRNA gene sequencing. Its AcOEt crude extract was successfully analyzed using non-targeted LC-MS/MS analysis, and molecular networking, leading to a putative annotation of its chemical diversity thanks to spectral libraries from GNPS and in s...

In this work, a novel series of racemic Schiff bases bearing the biologically active [1,2,4]triazolo[5,1-b]quinazoline scaffold has been synthesized in good to excellent yields, and fully characterized by FTIR, 1H NMR and 13C NMR, as well as elemental analyses. The in vitro antimalarial activities of these derivatives have been evaluated against Pl...

Small molecules that can restore the action of legacy antibiotics toward drug-resistant bacteria represent an area of ongoing research interest. We have previously reported indole-3-glyoxylamido and indole-3-acetamido-polyamine conjugates that exhibit intrinsic activity toward bacterial and fungal species, and the ability to enhance the action of d...

Many people around the world suffer from malaria, especially in tropical or subtropical regions. While malaria medications have shown success in treating malaria, there is still a problem with resistance to these drugs. Herein, we designed and synthesized some structurally novel benzotriazole‐β‐lactams using 2‐(1H‐benzo[d][1,2,3]triazol‐1‐yl)acetic...

Citation: Vergoz, D.; Le, H.; Bernay, B.; Schaumann, A.; Barreau, M.; Nilly, F.; Desriac, F.; Tahrioui, A.; Giard, J.-C.; Lesouhaitier, O.; et al. Antibiofilm and Abstract: The emergence of multi-drug resistant pathogens is a major public health problem, leading us to rethink and innovate our bacterial control strategies. Here, we explore the antib...

Citation: Sue, K.; Cadelis, M.M.; Hainsworth, K.; Rouvier, F.; Bourguet-Kondracki, M.-L.; Brunel, J.M.; Copp, B.R. Preliminary SAR of Novel Pleuromutilin-Polyamine Conjugates. Microorganisms 2023, 11, 2791. https://doi. Abstract: While pleuromutilin (1) and its clinically available derivatives (2-6) are highly effective against Gram-positive bacter...

In this work, two groups of structurally novel indolo-β-lactam hybrids were synthesized by ketene-imine [2 + 2] cycloaddition reaction. In the first series, the reaction proceeded between various selected aromatic imines and a ketene derived from indole-3-acetic acid. For the second, N-acetic acid indole-3-carbaldehyde was used as the ketene source...

The widespread incidence of antimicrobial resistance necessitates the discovery of new classes of antimicrobials as well as adjuvant molecules that can restore the action of ineffective antibiotics. Herein, we report the synthesis of a new class of indole-3-acetamido-polyamine conjugates that were evaluated for antimicrobial activities against a pa...

Introduction Cyanobacteria are important members of the dense biofilms that colonize available substrates in mangrove habitats worldwide. However, their taxonomic diversity and biological activities have received little attention. Methods The occurrence of cyanobacteria is evaluated in 27 biofilms collected from mangroves in Mayotte. Filamentous c...

The marine natural product ianthelliformisamine C is a bis-cinnamido substituted spermine derivative that exhibits intrinsic antimicrobial properties and can enhance the action of doxycycline towards the Gram-negative bacterium Pseudomonas aeruginosa. As part of a study to explore the structure–activity requirements of these activities, we have syn...

Citation: Cadelis, M.M.; Edmeades, L.R.; Chen, D.; Gill, E.S.; Fraser, K.; Rouvier, F.; Bourguet-Kondracki, M.-L.; Brunel, J.M.; Copp, B.R. Abstract: As part of our search for new antimicrobials and antibiotic enhancers, a series of naphthyl-and biphenyl-substituted polyamine conjugates have been synthesized. The structurally-diverse library of com...

Antibiotic resistance is a growing global health threat, requiring urgent attention. One approach to overcome antibiotic resistance is to discover and develop new antibiotic enhancers, molecules that work with legacy antibiotics to enhance their efficacy against resistant bacteria. Our previous screening of a library of purified marine natural prod...

Citation: Draveny, M.; Rose, C.; Pinet, A.; Ferrié, L.; Figadère, B.; Brunel, J.-M.; Masi, M. Scope and Limitations of Exploiting the Ability of the Chemosensitizer NV716 to Enhance the Activity of Tetracycline Derivatives against Pseudomonas aeruginosa. Molecules 2023, 28, 4262. Abstract: The spread of antibiotic resistance is an urgent threat to...

Citation: Dofini Magnini, R.; Pedinielli, F.; Vergalli, J.; Ouedraogo, N.; Remy, S.; Hilou, A.; Brunel, J.-M.; Pagès, J.-M.; Davin-Regli, A. Acacia senegal Budmunchiamines as a Potential Adjuvant for Rejuvenating Phenicol Activities towards Escherichia coli-Resistant Strains. Int. J. Mol. Sci. 2023, 24, 8790. https:// Abstract: The continuous emerg...

Citation: Vergoz, D.; Nilly, F.; Desriac, F.; Barreau, M.; Géry, A.; Lepetit, C.; Sichel, F.; Jeannot, K.; Giard, J.-C.; Garon, D.; et al. 6-Polyaminosteroid Squalamine Analogues Display Antibacterial Activity against Resistant Pathogens. Int. J. Mol. Sci. 2023, 24, 8568. https://doi. Abstract: A series of 6-polyaminosteroid analogues of squalamine...

Doxycycline is a tetracycline commonly used for its antibiotic properties and capacity to treat acne- and rosacea-like skin lesions. It has also recently demonstrated interesting effects against Parkinson's disease pathomechanisms. Notably, doxycycline was reported to limit amyloid-type aggregation of α-synuclein and curtail neurodegeneration-relat...

The Gram-positive bacterium Staphylococcus epidermidis is responsible for important nosocomial infections. With the continuous emergence of antibiotic-resistant strains, the search for new treatments has been amplified in the last decades. A potential candidate against multidrug-resistant bacteria is squalamine, a natural aminosterol discovered in...

In this study, α-ω-disubstituted polyamines exhibit a range of potentially useful biological activities, including antimicrobial and antibiotic potentiation properties. We have prepared an expanded set of diarylbis(thioureido)polyamines that vary in central polyamine core length, identifying analogues with potent methicillin-resistant Staphylococcu...

Several studies have reported that the tetracycline (TC) class antibiotic doxycycline (DOX) is effective against Parkinson’s disease (PD) pathomechanisms. The aim of the present work was three-fold: (i) Establish a model system to better characterize neuroprotection by DOX; (ii) Compare the rescue effect of DOX to that of other TC antibiotics; (iii...

With the increased incidence of antibiotic resistance, the discovery and development of new antibacterials is of increasing importance and urgency. The report of the natural product antibiotic squalamine in 1993 has stimulated a lot of interest in the study of structurally simplified cholic acid-polyamine derivatives. We report the synthesis of a f...

Citation: Tabcheh, J.; Vergalli, J.; Davin-Régli, A.; Ghanem, N.; Pages, J.-M.; Al-Bayssari, C.; Brunel, J.M. Abstract: Antibiotic resistance continues to evolve and spread beyond all boundaries, resulting in an increase in morbidity and mortality for non-curable infectious diseases. Due to the failure of conventional antimicrobial therapy and the...

Cryptococcus neoformans (Cn) is a pathogenic yeast that is the leading cause of fungal meningitis in immunocompromised patients. Various Cn virulence factors, such as the enzyme laccase and its product melanin, phospholipase, and capsular polysaccharide have been identified. During a screen of knockout mutants, the gene resistance to aminocholester...

Antibiotics have been the cornerstone of modern medicine saving lives by virtue of being able to cure infectious diseases and to prevent infections in those who are immune compromised. Their intense use has led to a surging increase in the incidence of antibiotic-resistant bacteria resulting in a desperate need for antibiotics with new mechanisms o...

Gram-negative bacteria were reported as a significant cause of infections in both community and nosocomial settings. Considered as one of the greatest threats to public health, the spread of bacteria drug resistance and the lack of effective alternative treatment options remains problematic. Herein, we report a promising strategy to combat Gram-neg...

New therapeutic options to combat the growing incidence of antimicrobial resistance are urgently needed. A 2015 publication reported the isolation and biological evaluation of two diketopiperazine natural products, cyclo(l-Trp-l-Arg) (CDP 2) and cyclo(d-Trp-d-Arg) (CDP 3), from an Achromobacter sp. bacterium, finding that the latter metabolite in p...

The antibiotic tetracycline demeclocycline (DMC) was recently reported to rescue α-synuclein (α-Syn) fibril-induced pathology. However, the antimicrobial activity of DMC precludes its potential use in long-term neuroprotective treatments. Here, we synthesized a doubly reduced DMC (DDMC) derivative with residual antibiotic activity and improved neur...

Pseudomonas aeruginosa is intrinsically resistant to many antibiotics due to the impermeability of its outer membrane and to the constitutive expression of efflux pumps. Here, we show that the polyaminoisoprenyl compound NV716 at sub-MIC concentrations re-sensitizes P. aeruginosa to abandoned antibiotics by binding to the lipopolysaccharides (LPS)...

The antibiotic tetracycline demeclocycline (DMC) was recently reported to rescue α-synuclein (α-Syn) fibril-induced pathology. However, the antimicrobial activity of DMC precludes its po-tential use in long-term neuroprotective treatments. Here, we synthesized a DMC derivative with residual antibiotic activity and improved neuroprotective effects....

In the present work, a novel series of monocyclic 1H‐phenanthro[9,10‐d]imidazole β‐lactam hybrids has been synthesized in yields varying from 70–95 % by a stereoselective [2+2]‐cyclocondensation (Staudinger reaction). The newly‐synthesized compounds and their imine intermediates were characterized by IR, ¹H NMR, ¹³C NMR and elemental analyses. The...

Aims The antibiotic tetracycline demeclocycline (DMC) was recently reported to rescue α-Synuclein (α-Syn) fibril-induced pathology (Braun et al., NPJ Parkinsons Dis, 2021). However, their antibiotic activity precludes its potential use for long-term neuroprotective treatments. Here, our aim was to synthesize a DMC derivative named DDMC with low ant...

In our ongoing search for bioactive compounds, a class of novel spiro-β-lactam isatin hybrids has been synthesized through a [2 + 2] cycloaddition reaction from 1-allyl-3-(arylimino)indolin-2-one, ketenes and various aryloxy acetic acids. The formation of all cycloadducts was confirmed by FTIR, ¹H NMR, ¹³C NMR, and mass spectroscopy as well as elem...

Potentiators can improve antibiotic activity against difficult-to-treat Gram-negative bacteria like Escherichia coli, Klebsiella pneumoniae or Acinetobacter baumannii. They represent an appealing strategy in view of the paucity of therapeutic alternatives in case of multidrug resistance. Here, we examine the ability of the polyamino-isoprenyl compo...

As part of our search for new antimicrobials and antibiotic adjuvants, a series of podocarpic acid-polyamine conjugates have been synthesized. The library of compounds made use of the phenolic and carboxylic acid moieties of the diterpene allowing attachment of polyamines (PA) of different lengths to afford a structurally-diverse set of analogues....

The lack of novel drugs in development and the combination of increased incidence of drug-resistant strains of bacteria has created the need for the search for new antimicrobials as well as new original strategies to fight bacterial resistance. In this context, a series of polyamine quinoline derivatives were prepared and biologically evaluated, id...

In our ongoing search for bioactive compounds, a class of novel spiro- β -lactam isatin hybrids have been obtained by [2+2] cycloaddition reaction from 1-allyl-3-(arylimino)indolin-2-one and ketenes from various aryloxy acetic acids. The formation of all cycloadducts were confirmed by FTIR, ¹ H NMR, ¹³ C NMR, and Mass spectra as well as elemental a...

A series of oleanolic acid derivatives holding oxo- or 3-N-polyamino-3-deoxy-substituents at C3 as well as carboxamide function at C17 with different long chain polyamines have been synthesized and evaluated for antimicrobial activities. Almost all series presented good to moderate activity against Gram-positive S. aureus, S. faecalis and B. cereus...

Poly(dimethyl siloxane) (PDMS) is one of the most widely used materials in the biomedical field, but due to its hydrophobic character it is prone to bacterial adhesion and biofilm formation. Prevention of bacterial adhesion by contact-killing surface is one of the promising strategies, although bacterial resistance to conventional active molecules...

The design and evaluation of antimicrobial activities of an isonitrosomalononitrile silver(I) salt was reported. This highly stable water-soluble silver salt shows Minimum Inhibitory Concentrations (MIC) values ranging from 0.15 to 5 µg/mL towards both sensitive and resistant Gram-positive and negative bacteria. Furthermore, this silver salt has be...

Essential oils (EOs) or their components are widely used by inhalation or nebulization to fight mild respiratory bacterial infections. However, their interaction with antibiotics is poorly known. In this study we evaluated the effects of citral, the main component of lemongrass oil, on in vitro susceptibility of Pseudomonas aeruginosa to antibiotic...

Several monocyclic β‐lactams have been synthesized via a [2+2] ketene–imine cycloaddition reaction (Staudinger reaction) and evaluated for their biological activities. The structure of synthesized products was confirmed by spectral data and elemental analyses. β‐Lactams 4 b and 4 h exhibited 31 and 27 anti‐inflammatory ratios, respectively, which a...

The cleavage of the insulin receptor by β-secretase 1 (BACE1) in the liver increases during diabetes, which contributes to reduce insulin receptor levels and impair insulin signaling. However, the precise signaling events that lead to this increased cleavage are unclear. We showed that BACE1 cleaves the insulin receptor in the early secretory pathw...

In our search for new antibiotic adjuvants as a novel strategy to deal with the emergence of multi-drug resistant (MDR) bacteria, a series of succinylprimaquine-polyamine (SPQ-PA) conjugates and derivatives of a cationic amphiphilic nature have been prepared. Evaluation of these primaquine conjugates for intrinsic antimicrobial properties and the a...

Active efflux confers intrinsic resistance to multiple antibiotics in Pseudomonas aeruginosa , including old disused molecules. Beside resistance, intracellular survival is another reason for failure to eradicate bacteria with antibiotics. We evaluated the capacity of polyaminoisoprenyl potentiators (designed as efflux pump inhibitors [EPIs]) NV716...

The outer membrane (OM) of Gram-negative bacteria provides an efficient barrier against external noxious compounds such as antimicrobial agents. Associated with drug target modification, it contributes to the overall failure of chemotherapy. In the complex OM architecture, Lipid A plays an essential role by anchoring the lipopolysaccharide in the m...

The global increase of multidrug resistant bacteria and the lack of new classes of antibiotic especially those targeting Gram-negative pathogens are leaving the clinicians disarmed to treat numerous bacterial infections. Recently, the design of adjuvants able to enhance antibiotics activities appears to be one of the most promising investigated sol...

An improved, fast, and accurate procedure for regioselective microwave Oppenauer oxidation of sterol derivatives is reported leading to the expected corresponding ketosterols in moderate to excellent isolated yields ranging from 58 to 81%. The influence of different parameters dealing with the nature of the solvent and of the Lewis acid/ketone coup...

The growing number of multidrug resistant strains in Tunisia has become a serious health concern contributing to high rate of mortality and morbidity. Since current antibiotics are rapidly becoming ineffective, novel strategies to combat resistance are needed. Recently, we demonstrated that combination of a tetracycline antibiotic with various poly...

New antibiotics are urgently needed to address the mounting resistance challenge. In early drug discovery one of the bottlenecks is the elucidation of targets and mechanisms. To accelerate antibiotic research, we provide a proteomic approach for the rapid classification of compounds into those with precedented and unprecedented modes of action. We...

Today, an alarming rise of bacterial gastroenteritis in humans resulting from consuming Campylobacter-tainted foods is being observed. One of the solutions for mitigating this issue may be the antibacterial activity of essential oils. In the present research, we propose to study the antibacterial activity against Campylobacter and other Gram-negati...

A proprietary library of novel N-aryl-substituted amino acid derivatives bearing a hydroxamate head group allowed the identification of compound 3a that possesses weak proadipogenic and peroxisome proliferator-activated receptor γ (PPARγ) activating properties. The systematic optimization of 3a, in order to improve its PPARγ agonist activity, led t...

Recently, extensive researches have emphasized the fact that polyamines conjugates are becoming important in all the biological and medicinal fields. In this review we will focus our attention on natural polyamines and highlight recent progress in both fundamental mechanism studies and interest for the development and application for a therapeutic...

Today, an alarming rise of bacterial gastroenteritis in humans resulting from consuming Campylobacter-tainted foods is being observed. One of the solutions for mitigating this issue may be the antibacterial activity of essential oils. In the present research, we propose to study the antibacterial activity against Campylobacter and other Gram-negati...

The discovery of new antibiotic adjuvants is an attractive option for overcoming antimicrobial resistance. We have previously reported the discovery of a bis‐6‐bromoindolglyoxylamide derivative of spermine as being able to enhance the action of antibiotics against Gram‐negative bacteria but suffers from being cytotoxic and red‐blood cell haemolytic...

A series consisting of new polyaminoisoprenyl derivatives were prepared in moderate to good chemical yields varying from 32 to 64% according two synthetic pathways: 1) using a titanium reductive amination reaction affording a 50/50 mixture of cis and trans isomers 2) a direct nucleophilic substitution leading to a stereoselective synthesis of the c...

Peptidoglycan (PG) is an essential polymer of the bacterial cell wall and a major antibacterial target. Its synthesis requires glycosyltransferase (GTase) and transpeptidase enzymes that, respectively, catalyze glycan chain elongation and their cross-linking to form the protective sacculus of the bacterial cell. The GTase domain of bifunctional pen...

The rise of antimicrobial resistance has created an urgent need for the development of new methods for antibiotics delivery to patients with pulmonary infections in order to mainly increase the effectiveness of the drugs administration, to minimize the risk of emergence of resistant strains, and to prevent patients reinfection. Since bacterial resi...

This study reports the synthesis and biological investigation of three series of novel monocyclic β-lactam derivatives bearing a morpholine ring substituent on the nitrogen. The resulting β-lactam adducts were synthesized via Staudinger's [2+2]-ketene-imine cycloaddition reaction. New synthesized products were fully characterized by spectral data a...

Introduction An attractive antibiotic-adjuvant strategy consisting in the design and synthesis of polyaminoisoprenyl molecules able to restore antibiotic activity of tetracycline antibiotics against resistant Pseudomonas aeruginosa bacterial strains has been developed. Methods These chemo-sensitizers are readily prepared from geraniol and farnesol...

The lack of novel classes of antibiotics as well as the constant increase of multidrug resistant bacteria are leaving the clinicians disarmed to treat bacterial infections, especially those caused by Gram-negative pathogens. Among all the investigated solutions, the design of adjuvants able to enhance antibiotics activities appears to be one of the...

PPARγ represents a key target for the treatment of type II diabetes and metabolic syndrome. Synthetic antidiabetic drugs activating PPARγ are accompanied by serious undesirable side effects related to their agonism. In the search for new PPARγ regulators, inhibitors of PPARγ phosphorylation on S245 mediated by CDK5 represent an opportunity for the...

NV669 is an aminosterol derived from squalamine found to possess strong anticancer effects. The aim of this study was to investigate NV669’s beneficial effects on human pancreatic and hepatic cancer models and to decipher the cellular and molecular mechanisms involved in tumor growth decrease upon treatment with NV669. Pancreatic (BxPC3, MiaPaCa-2)...

Synthesis of A-ring-modified lupane, oleanane and ursane type triterpenoid conjugates with spermidine through an aldimine linkage or diethylentriamine via an amide bond is described. These derivatives were evaluated for their in vitro antimicrobial properties against human pathogens. Except for derivatives 1 and 7, all compounds have moderate to we...

A series of substituted di-indolglyoxylamido-spermine analogues were prepared and evaluated for intrinsic antimicrobial properties and the ability to enhance antibiotic action. As a compound class, intrinsic activity was typically observed towards Gram-positive bacteria and the fungus Cryptococcus neoformans, with notable exceptions being the 5-bro...

The spreading of multidrug-resistant bacteria and the lack of novel antibiotic molecules leave clinicians and veterinarians with very limited options to treat bacterial infections, especially those caused by Gram-negative pathogens. To reduce the selection of antibiotic resistance mechanisms and their transfer to human pathogens, veterinary pharmac...

Background and Objective: Nasal carriage of Staphylococcus aureus (S. aureus) constitutes an important risk factor for subsequent infections in some types of patient populations. Decolonization of carriers using intranasal mupirocin is widely used as a preventive measure. However, resistance to this agent has been rising and causing failure in the...

Highly diastereoselective synthesis of chromeno β-lactam hybrids was achieved by an efficient one-pot three-component reaction. With this procedure, the desired β-lactam products were obtained in good yields and with exclusive cis stereoselection, by combining a variety of benzaldehydes, malononitrile, and either 5,5-dimethylcyclohexane-1,3-dione o...

Multiple approaches have been developed to combat bacterial resistance. However, the combination of antibiotic resistance mechanisms by bacteria and the limited number of effective antibiotics available, decreases the effective interventions for the treatment of current bacterial infections. This review covers the many ways that bacteria resist ant...

WHO considers P. aeruginosa (PA) as a priority pathogen for the search of innovative therapies. PA is indeed intrinsically resistant to many antibiotics due to poor outer membrane permeability and/or active efflux [1]. Moreover, it can also adopt specific lifestyles, like intracellular survival, which are poorly responsive to antibiotics [2]. Our a...

The combination of increased incidence of drug-resistant strains of bacteria and a lack of novel drugs in development creates an urgency for the search for new antimicrobials. Initial screening of compounds from an in-house library identified two 6-bromoindolglyoxylamide polyamine derivatives (3 and 4) that exhibited intrinsic antimicrobial activit...

Fifteen novel β-lactams bearing N-ethyl tert-butyl carbamate group 5a-o and fifteen N-(2- aminoethyl) β-lactams 6a-o were synthesized by [2+2] ketene-imine cycloaddition reaction (Staudinger). The cycloaddition reaction was found to be totally diastereoselective leading exclusively to theformation of cis-β-lactam derivatives. These newly synthesize...

Squalamine is a natural polycationic aminosterol extracted from the shark Squalus acanthias . Squalamine displays remarkable efficacy against antimicrobial-resistant Gram-negative and Gram-positive bacteria. Its membranolytic activity and low cytotoxicity make squalamine one of the most promising agents to fight nosocomial pathogens such as Acineto...

Some new β-lactams bearing biologically important morpholine ring have been synthesized by acylation of amino β-lactams in the presence of morpholine-4-carbonyl chloride. These novel β-lactams were prepared under mild reaction conditions without any solvent in short reaction times. Their biological activities have been examined against microbial ag...

Background: Squalamine is a natural polycationic aminosterol, extracted from Squalus acanthias. It displays a remarkable efficacy against Gram- and Gram+ bacteria which are resistant to antimicrobials. Its membranolytic activity and its low cytotoxicity make squalamine, one of the most promising agents to fight against nosocomial pathogens such as...

On the basis of a recent article "Predicting reaction performance in C-N cross-coupling using machine learning" that appeared in Science, we had decided to highlight the way forward for artificial intelligence in chemistry. Synthesis of molecules remains one of the most important challenges in organic chemistry, and the standard approach involved b...

In an effort to explore the antibacterial potential of the marine natural product halocyamine A, a series of analogues including desbromo and alanine-substituted variants were synthesised and evaluated for biological activity against a panel of Gram-positive and –negative bacteria. The analogues were synthesised by a combination of solid-phase pept...

In HER2-positive breast cancer, the Trastuzumab + Docetaxel doublet has a significant efficacy but is hampered by frequent toxicities that could be addressed with nanoparticles. We have developed an Antibody-Nano Conjugate (ANC), combining docetaxel encapsulated in a stealth liposome engrafted with trastuzumab. This entity is expected to improve th...

Background Trastuzumab plus docetaxel is a mainstay to treat HER2-positive breast cancers. However, developing nanoparticles could help to improve the efficacy/toxicity balance of this doublet by improving drug trafficking and delivery to tumors. This project aimed to develop an immunoliposome in breast cancer, combining docetaxel encapsulated in a...

TEM observations of ANC-1 treated with osmium tetroxide (A and B) and uranyl acetate (C). Abbreviations: TEM, transmission electron microscopy; ANC-1, antibody nanoconjugate-1.

TEM observations of liposome-1 before extrusion. Abbreviation: TEM, transmission electron microscopy.

Representative SDS-PAGE gel, showing trastuzumab engraftment on liposome surface. Protein markers (A1), trastuzumab (A2), thiolated trastuzumab (A3), Liposome-1 (A4), ANC-1 engrafted with strategy B (A5), Liposome-1 engrafted with strategy B without maleimide function (A6), ANC-1 engrafted following strategy A (A7), Liposome-1 engrafted following s...