This prospective study was designed to establish whether vascular endoscopy would provide more in... more This prospective study was designed to establish whether vascular endoscopy would provide more information on the graft lumen than standard completion angiography during infrainguinal bypass surgery. Ninety-nine patients with 102 infragenicular bypass grafts who underwent both angiography and angioscopy intraoperatively were evaluated. In 99 of the 102 patients the indication was critical limb ischaemia. Of the 102 bypass grafts, 81 were autogenous vein. Distally, 24 grafts were anastomosed to the below-knee popliteal segment, 64 extended to the crural and 14 to the pedal arteries. On completion of the distal anastomosis, grafts were first evaluated by angiography and then by angioscopy. The images obtained with the two monitoring modalities were compared by the operating surgeon and re-explorations were performed immediately if necessary. Completion angiography and angioscopy produced images of good quality in 96 and 97 cases, respectively. In 12 cases completion angiography showed abnormalities. Of these, five were located below the distal anastomosis and were not accessible to angioscopic examination. Conduit defects were found in seven instances. In one of them angioscopy showed the angiogram to be false-positive. Of the 90 grafts with normal completion angiograms, seven were found to show significant pathology on angioscopy. Compared to angioscopy, the sensitivity and specificity of angiography to detect abnormalities within the graft was 46% and 98%, respectively. Our results suggest that vascular endoscopy is superior to angiography for disclosing conduit defects, but that it does not provide adequate information about the distal arterial anatomy.
Potential approaches to achieve cytosolic delivery of liposomal macromolecules are presented. The... more Potential approaches to achieve cytosolic delivery of liposomal macromolecules are presented. These approaches include: 1) the co-encapsulation of fusogenic peptides into targeted drug-containing liposomes, 2) coupling of the HIV-1 derived cell penetrating peptide TAT to the surface of liposomes and 3) photochemical internalisation, based on photochemically inducible permeabilisation of endocytic vesicles
Nucleic acid based therapeutics are currently being studied for their application in cancer thera... more Nucleic acid based therapeutics are currently being studied for their application in cancer therapy. In this study, the effect of different cationic delivery systems on the circulation kinetics, tumor localization, and tissue distribution of short interfering RNA (siRNA) and plasmid DNA (pDNA) was examined, after intravenous administration in mice bearing a s.c. Neuro 2A tumor. Nanosized particles were formed upon complexation of siRNA with the cationic liposome formulation DOTAP/DOPE and the targeted, cationic polymer RGD-PEG-PEI. Both the circulation kinetics and the overall tumor localization of the siRNA complexes were similar to non-complexed siRNA. Importantly, the different carriers changed the intratumoral distribution of siRNA within the tumor. pDNA was effectively condensed with linear polyethylenimine (PEI), PEGylated linear PEI (PEG-PEI) or poly(2-dimethylamino ethylamino)phosphazene. Only PEG-PEI was able to improve the pDNA circulation kinetics. All pDNA complexes yielded similar pDNA tumor localization (1% of the injected dose, 60 min after administration). We conclude that the level of nucleic acid tumor localization is independent on the type of formulation used in this study. Therefore, the value of carrier systems for the intravenous delivery of nucleic acids cannot be solely attributed to benefits relevant during the transport towards the tumor. Rather, the benefits are arising from carrier-induced changes in the intratumoral fate of the nucleic acids.
Carbon nanotubes (CNTs) have been studied for drug, antigen and nucleic acid delivery both in vit... more Carbon nanotubes (CNTs) have been studied for drug, antigen and nucleic acid delivery both in vitro and in vivo. Due to their nano-needle structure, they are supposed to cross the plasma membrane and enter directly into the cytoplasm likely upon an endocytosis-independent mechanism without inducing cell death. In this study, two cationically functionalized CNTs (CNT-PEI and CNT-pyridinium) were investigated for siRNA delivery. Both functionalized CNTs complexed siRNA and showed 10-30% silencing activity and a cytotoxicity of 10-60%. However, in terms of reduced toxicity or increased silencing activity, CNT-PEI and CNT-pyridinium did not show an added value over PEI and other standard transfection systems. Probably, the type of functionalization of carbon nanotubes might be a key parameter to obtain an efficient and non-cytotoxic CNT-based delivery system. Nevertheless, in view of the present results and importantly also of the non-degradability of CNTs, preference should currently be given to designing biodegradable carriers which mimic the needle structure of CNTs.
Liposome-encapsulated corticosteroids have shown to exert strong beneficial effects in inflammato... more Liposome-encapsulated corticosteroids have shown to exert strong beneficial effects in inflammatory diseases, such as arthritis and cancer. To extend the clinical applicability of these potent nanomedicines, the therapeutic effect of dexamethasone phosphate loaded long-circulating liposomes (LCL-DXP) was evaluated in animal models of multiple sclerosis (MS) and Crohn's disease (CD). In mice with experimental autoimmune encephalitis (EAE), a model for MS, treatment with LCL-DXP, but not free DXP, resulted in a decrease in disease activity when compared to PBS treated mice. In contrast, in mice with chronic DSS-induced colitis, a model for CD, treatment with LCL-DXP did not induce an improvement, but in fact worsened the fecal blood loss after treatment, indicating an aggravation of the disease. It is hypothesized that modulation of macrophage polarization towards a M2 phenotype underlies the efficacy of corticosteroid-based drug delivery systems, which is supported by the presented data. On the one hand, M1 polarized macrophages are part of the pathogenesis of MS; the modulation to M2-polarization by LCL-DXP is therefore beneficial. On the other hand, M1-polarized intestinal macrophages fulfill a protective and inflammation-suppressing role in intestinal homeostasis; changing their phenotype to M2 causes reduced protection to invading microorganisms, leading to a more severe intestinal inflammation. These findings therefore indicate that the interplay between the specific phenotype of macrophages and the specific inflammatory context of the inflammatory disease in question may be an important determining factor in the therapeutic applicability of liposomal corticosteroids in inflammatory disease.
Dendritic cells (DCs) are key antigen presenting cells that have the unique ability to present an... more Dendritic cells (DCs) are key antigen presenting cells that have the unique ability to present antigens on MHC molecules, which can lead to either priming or suppression of T cell mediated immune responses. C-type lectin receptors expressed by DCs are involved in antigen uptake and presentation through recognition of carbohydrate structures on antigens. Here we have explored the feasibility of modification of liposomes with glycans for targeting purposes to boost immune responses. The potential of targeting glycoliposomal constructs to the C-type lectin DC-SIGN on DCs was studied using either PEGylated or non-PEGylated liposomes. Our data demonstrate that formulation of the glycoliposomes as PEGylated negatively affected their potential to target to DCs.
The main constituent of bone is hydroxyapatite (HAP). Since HAP is only present in 'hard&... more The main constituent of bone is hydroxyapatite (HAP). Since HAP is only present in 'hard' tissues like bone and teeth, it represents a promising target for the selective drug delivery to bone. Due to the exceptional affinity of bisphosphonates (BP) for HAP, cholesteryl-trisoxyethylene-bisphosphonic acid (CHOL-TOE-BP), a new tailor-made BP derivative, was used as bone targeting moiety for liposomes. CHOL-TOE-BP-targeted liposomes were designed for the treatment of bone-related diseases to achieve prolonged local exposure to high concentrations of the bioactive compounds, thereby enhancing therapeutic efficacy and minimizing systemic side effects. The CHOL-TOE-BP-targeted liposomes were characterized regarding particle size and zeta potential. To study the bone targeting potential of these conjugates, an in vitro HAP binding assay was established. The obtained binding data indicate that CHOL-TOE-BP is useful as targeting device for liposomal drug delivery to bone.
The potential and limitations of targeted delivery of anticancer agents with colloidal particulat... more The potential and limitations of targeted delivery of anticancer agents with colloidal particulate carriers is the subject of this contribution. Because over the years liposomes have gained the most attention as carrier system in the category of colloidal carrier systems, this paper focuses on the utility of the liposomal system for tumor targeting. It is imperative that an intended therapeutic application of liposomes should be well matched with the liposome behavior in vivo. Therefore, the in vivo fate of the first-generation liposomes and the more recently developed second-generation liposomes (surface-modified liposomes such as the immunoli¬ posomes and long-circulating liposomes) is analyzed in terms of accessibility of target sites, time-, and sitecontrolled drug release and potential target sites for rational targeted delivery are discussed. A few examples of areas in cancer chemotherapy, with a strong rationale for the use of liposomes, are given. It is concluded that, although several options are available on the drawing board, issues such as tumor cell heterogeneity, ac¬ cess to the target site, shedding of antigens, and target site-specific release of the liposome-associated drug need to be addressed early in the development process.
Mesna is used as a mucolytic agent in CO, PD* and CF-patients and to prevent chemical cystitis in... more Mesna is used as a mucolytic agent in CO, PD* and CF-patients and to prevent chemical cystitis in cancer patients after nigh dose cyclophosphamide of ifosfamide. In urological patients with an" Indianapouch" or" Brickerbladder" mucus formadon is sometimes a ...
Macromolecular carrier materials based on N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic... more Macromolecular carrier materials based on N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic and well-characterized drug delivery systems that have been extensively evaluated in the past two decades, both at the preclinical and at the clinical level. Using several different imaging agents and techniques, HPMA copolymers have been shown to circulate for prolonged periods of time, and to accumulate in tumors both effectively and selectively by means of the Enhanced Permeability and Retention (EPR) effect. Because of this, HPMA-based macromolecular nanotheranostics, i.e. formulations containing both drug and imaging agents within a single formulation, have been shown to be highly effective in inducing tumor growth inhibition in animal models. In patients, however, as essentially all other tumor-targeted nanomedicines, they are generally only able to improve the therapeutic index of the attached active agent by lowering its toxicity, and they fail to improve the efficacy of the intervention. Bearing this in mind, we have recently reasoned that because of their biocompatibility and their beneficial biodistribution, nanomedicine formulations might be highly suitable systems for combination therapies. In the present manuscript, we briefly summarize several exemplary efforts undertaken in this regard in our labs in the past couple of years, and we show that long-circulating and passively tumor-targeted macromolecular nanotheranostics can be used to improve the efficacy of radiochemotherapy and of chemotherapy combinations.
To assist intravenously administered anticancer agents in achieving proper circulation times and ... more To assist intravenously administered anticancer agents in achieving proper circulation times and tumor concentrations, and to thereby improve the balance between their efficacy and their toxicity, a large number of drug delivery systems have been designed and evaluated over the years. Clinically relevant examples of such nanometer-sized carrier materials are liposomes, polymers, micelles and antibodies. In the vast majority of cases, however, and especially in patients, nanomedicine formulations are only able to attenuate the toxicity of the conjugated or entrapped chemotherapeutic drug, and they generally fail to improve the efficacy of the intervention. To overcome this shortcoming, and to broaden the clinical applicability of tumor-targeted nanomedicines, in the past 5 years we have developed several concepts for using N-(2-hydroxypropyl)methacrylamide (HPMA)-based polymer therapeutics to enhance the efficacy of combined modality anticancer therapy. Regarding surgery, HPMA copoly...
Second Congress of the European Society for Clinical Neuropharmacology Wtirzburg, November 9-11, ... more Second Congress of the European Society for Clinical Neuropharmacology Wtirzburg, November 9-11, 1995
A folate-poly(ethylene glycol) conjugate capable of covalent coupling to primary amines present a... more A folate-poly(ethylene glycol) conjugate capable of covalent coupling to primary amines present at the surface of polyplexes was developed. Coating of poly(dimethylaminomethyl methacrylate (pDMAEMA)-based polyplexes with this folate-pEG conjugate led to a sharp decrease of the z-potential, and a small increase in particle size. The size of the particles in isotonic medium did not change markedly in time demonstrating that rather stable particles were formed. The in vitro cellular toxicity of the pEGylated polyplexes with and without folate ligands was lowered considerably compared to uncoated polyplexes. The toxicity observed for the targeted pEGylated polyplexes was slightly higher than that of corresponding untargeted polyplexes, which might indicate an increased cellular association of targeted polyplexes. Transfection of OVCAR-3 cells in vitro was markedly increased compared to untargeted pEGylated polyplexes, suggesting targeted gene delivery.
A major limiting factor in the full exploitation of therapeutically active peptide and protein dr... more A major limiting factor in the full exploitation of therapeutically active peptide and protein drugs prepared synthetically and/ or by recombinant DNA technology, is the lack of availability of appropriate delivery systems. This contribution deals with the potential of liposomes for improved delivery of these drugs. Our current efforts in this field are briefly described: (1) longcirculating liposomes for prolonged systemic delivery of peptide drugs; (2) liposomes as carriers of interleukin-2 in locoregional anticancer immunotherapy; (3) plasminogen-bearing liposomes for the targeted delivery of tissue plasminogen activator; (4) immunoliposomes for the targeted delivery of diphtheria toxin; and (5) immunoliposomes bearing enzymes (immunoenzymosomes) for site-specific activation of prodrugs.
Understanding the role of the epidermal growth factor receptor (EGFR) in cellular signalling proc... more Understanding the role of the epidermal growth factor receptor (EGFR) in cellular signalling processes underlying malignancy has enabled the development of rationally designed EGFR-targeted therapeutics. Strategies have been devised to interfere with the EGFR signalling at three different levels: at the extracellular level, competing with ligand binding; at the intracellular level, inhibiting the activation of the tyrosine kinase; or at the mRNA level, modulating the expression of the EGFR protein. Each of these strategies has proven to have an antitumour effect mediated by events such as inhibition of cell proliferation, induction of apoptosis, decrease of cellular invasion and migration; and/or inhibition of angiogenesis. Furthermore, the combination of these strategies with traditional chemotherapy or radiotherapy has generally resulted in enhanced antitumour effects. Likewise, the benefit of interfering simultaneously with different signalling pathways has been documented to improve tumour growth inhibition. These preclinical results have encouraged clinical studies that led to the FDA approval of three drugs. However, finding the perfect strategy for each individual patient appears to be a limiting factor, demanding further research to be able to generate relevant molecular expression profiles on a case-to-case basis. Taken together, a successful EGFR inhibition will require a better understanding of signalling pathways in combination with the development of rationally designed effective molecules.
Objective. Angiogenic endothelial cells can function as phagocytes, and phagocytosis is initiated... more Objective. Angiogenic endothelial cells can function as phagocytes, and phagocytosis is initiated via the opsonin lactadherin. In this study, we examined the interaction between lactadherin-opsonized erythrocytes with reduced deformability and angiogenic endothelium, as loss of deformability is characteristic for suicidal and aged erythrocytes. Materials and Methods. We used the Arg-Gly-Asp (RGD)-modified erythrocyte model and investigated the deformability parameter by cross-linking erythrocyte membranes through treatment with glutaraldehyde. Association in vitro with primary endothelial cells was detected by flow cytometry and visualized by light, fluorescent, and electron microscopy. Involvement of two crucial factors in phagocytosis, a v-integrins and Rho guanosine triphosphatase family member Rac1, was studied using small interfering RNA technology. Modified erythrocytes were administered in vivo into tumor-bearing mice to detect phagocytosis by endothelial cells. Results. Glutaraldehyde-treated (rigid) RGD-modified erythrocytes showed a strongly enhanced endothelial cell association compared to flexible RGD-modified erythrocytes. Knockdown by small interfering RNA lipoplexes of a v-integrins and Rac1 confirmed classical tethering and internalization of rigid RGD-erythrocytes. Upon in vivo administration, tumor endothelium showed pronounced erythrophagocytosis. Conclusion. The pronounced phagocytosis of opsonized erythrocytes with reduced deformability by angiogenic growth factorLactivated endothelial cells evokes new insights in endothelial cell function and suggests a role for these endothelial cells in (hematological) disorders because of their capacity to clear disordered erythrocytes.
In the course of an inflammation, vascular endothelial cells (VECs) are strongly involved in proc... more In the course of an inflammation, vascular endothelial cells (VECs) are strongly involved in processes like leukocyte recruitment, cytokine production, and angiogenesis. Specific interference in these processes may yield great therapeutic benefit in the treatment of (chronic) inflammatory disorders. Drug targeting to VECs at inflamed sites may allow such intervention. VECs at inflamed sites represent a very well-accessible target cell population for circulating drug-targeting systems, which may also be selectively distinguished from normal VECs by the expression of several cell surface receptors involved in the inflammation. One group of specifically expressed molecules are the adhesion molecules (AMs), which have a major function in adhesion of cells to each other, to the extracellular matrix, or in the adhesion and subsequent recruitment of circulating immune cells. This review describes AMs with regard to their function in the inflammatory disease and their usefulness in functioning as a specific target receptor for drug-targeting approaches in general and with an emphasis on liposome-based drug delivery.
Reproduce material from Drug Discovery Today? This publication and the contributions it contains ... more Reproduce material from Drug Discovery Today? This publication and the contributions it contains are protected by the copyright of Elsevier. Except as outlined in the terms and conditions (see p. X), no part of this journal can be reproduced without written permission from Elsevier,
This prospective study was designed to establish whether vascular endoscopy would provide more in... more This prospective study was designed to establish whether vascular endoscopy would provide more information on the graft lumen than standard completion angiography during infrainguinal bypass surgery. Ninety-nine patients with 102 infragenicular bypass grafts who underwent both angiography and angioscopy intraoperatively were evaluated. In 99 of the 102 patients the indication was critical limb ischaemia. Of the 102 bypass grafts, 81 were autogenous vein. Distally, 24 grafts were anastomosed to the below-knee popliteal segment, 64 extended to the crural and 14 to the pedal arteries. On completion of the distal anastomosis, grafts were first evaluated by angiography and then by angioscopy. The images obtained with the two monitoring modalities were compared by the operating surgeon and re-explorations were performed immediately if necessary. Completion angiography and angioscopy produced images of good quality in 96 and 97 cases, respectively. In 12 cases completion angiography showed abnormalities. Of these, five were located below the distal anastomosis and were not accessible to angioscopic examination. Conduit defects were found in seven instances. In one of them angioscopy showed the angiogram to be false-positive. Of the 90 grafts with normal completion angiograms, seven were found to show significant pathology on angioscopy. Compared to angioscopy, the sensitivity and specificity of angiography to detect abnormalities within the graft was 46% and 98%, respectively. Our results suggest that vascular endoscopy is superior to angiography for disclosing conduit defects, but that it does not provide adequate information about the distal arterial anatomy.
Potential approaches to achieve cytosolic delivery of liposomal macromolecules are presented. The... more Potential approaches to achieve cytosolic delivery of liposomal macromolecules are presented. These approaches include: 1) the co-encapsulation of fusogenic peptides into targeted drug-containing liposomes, 2) coupling of the HIV-1 derived cell penetrating peptide TAT to the surface of liposomes and 3) photochemical internalisation, based on photochemically inducible permeabilisation of endocytic vesicles
Nucleic acid based therapeutics are currently being studied for their application in cancer thera... more Nucleic acid based therapeutics are currently being studied for their application in cancer therapy. In this study, the effect of different cationic delivery systems on the circulation kinetics, tumor localization, and tissue distribution of short interfering RNA (siRNA) and plasmid DNA (pDNA) was examined, after intravenous administration in mice bearing a s.c. Neuro 2A tumor. Nanosized particles were formed upon complexation of siRNA with the cationic liposome formulation DOTAP/DOPE and the targeted, cationic polymer RGD-PEG-PEI. Both the circulation kinetics and the overall tumor localization of the siRNA complexes were similar to non-complexed siRNA. Importantly, the different carriers changed the intratumoral distribution of siRNA within the tumor. pDNA was effectively condensed with linear polyethylenimine (PEI), PEGylated linear PEI (PEG-PEI) or poly(2-dimethylamino ethylamino)phosphazene. Only PEG-PEI was able to improve the pDNA circulation kinetics. All pDNA complexes yielded similar pDNA tumor localization (1% of the injected dose, 60 min after administration). We conclude that the level of nucleic acid tumor localization is independent on the type of formulation used in this study. Therefore, the value of carrier systems for the intravenous delivery of nucleic acids cannot be solely attributed to benefits relevant during the transport towards the tumor. Rather, the benefits are arising from carrier-induced changes in the intratumoral fate of the nucleic acids.
Carbon nanotubes (CNTs) have been studied for drug, antigen and nucleic acid delivery both in vit... more Carbon nanotubes (CNTs) have been studied for drug, antigen and nucleic acid delivery both in vitro and in vivo. Due to their nano-needle structure, they are supposed to cross the plasma membrane and enter directly into the cytoplasm likely upon an endocytosis-independent mechanism without inducing cell death. In this study, two cationically functionalized CNTs (CNT-PEI and CNT-pyridinium) were investigated for siRNA delivery. Both functionalized CNTs complexed siRNA and showed 10-30% silencing activity and a cytotoxicity of 10-60%. However, in terms of reduced toxicity or increased silencing activity, CNT-PEI and CNT-pyridinium did not show an added value over PEI and other standard transfection systems. Probably, the type of functionalization of carbon nanotubes might be a key parameter to obtain an efficient and non-cytotoxic CNT-based delivery system. Nevertheless, in view of the present results and importantly also of the non-degradability of CNTs, preference should currently be given to designing biodegradable carriers which mimic the needle structure of CNTs.
Liposome-encapsulated corticosteroids have shown to exert strong beneficial effects in inflammato... more Liposome-encapsulated corticosteroids have shown to exert strong beneficial effects in inflammatory diseases, such as arthritis and cancer. To extend the clinical applicability of these potent nanomedicines, the therapeutic effect of dexamethasone phosphate loaded long-circulating liposomes (LCL-DXP) was evaluated in animal models of multiple sclerosis (MS) and Crohn's disease (CD). In mice with experimental autoimmune encephalitis (EAE), a model for MS, treatment with LCL-DXP, but not free DXP, resulted in a decrease in disease activity when compared to PBS treated mice. In contrast, in mice with chronic DSS-induced colitis, a model for CD, treatment with LCL-DXP did not induce an improvement, but in fact worsened the fecal blood loss after treatment, indicating an aggravation of the disease. It is hypothesized that modulation of macrophage polarization towards a M2 phenotype underlies the efficacy of corticosteroid-based drug delivery systems, which is supported by the presented data. On the one hand, M1 polarized macrophages are part of the pathogenesis of MS; the modulation to M2-polarization by LCL-DXP is therefore beneficial. On the other hand, M1-polarized intestinal macrophages fulfill a protective and inflammation-suppressing role in intestinal homeostasis; changing their phenotype to M2 causes reduced protection to invading microorganisms, leading to a more severe intestinal inflammation. These findings therefore indicate that the interplay between the specific phenotype of macrophages and the specific inflammatory context of the inflammatory disease in question may be an important determining factor in the therapeutic applicability of liposomal corticosteroids in inflammatory disease.
Dendritic cells (DCs) are key antigen presenting cells that have the unique ability to present an... more Dendritic cells (DCs) are key antigen presenting cells that have the unique ability to present antigens on MHC molecules, which can lead to either priming or suppression of T cell mediated immune responses. C-type lectin receptors expressed by DCs are involved in antigen uptake and presentation through recognition of carbohydrate structures on antigens. Here we have explored the feasibility of modification of liposomes with glycans for targeting purposes to boost immune responses. The potential of targeting glycoliposomal constructs to the C-type lectin DC-SIGN on DCs was studied using either PEGylated or non-PEGylated liposomes. Our data demonstrate that formulation of the glycoliposomes as PEGylated negatively affected their potential to target to DCs.
The main constituent of bone is hydroxyapatite (HAP). Since HAP is only present in 'hard&... more The main constituent of bone is hydroxyapatite (HAP). Since HAP is only present in 'hard' tissues like bone and teeth, it represents a promising target for the selective drug delivery to bone. Due to the exceptional affinity of bisphosphonates (BP) for HAP, cholesteryl-trisoxyethylene-bisphosphonic acid (CHOL-TOE-BP), a new tailor-made BP derivative, was used as bone targeting moiety for liposomes. CHOL-TOE-BP-targeted liposomes were designed for the treatment of bone-related diseases to achieve prolonged local exposure to high concentrations of the bioactive compounds, thereby enhancing therapeutic efficacy and minimizing systemic side effects. The CHOL-TOE-BP-targeted liposomes were characterized regarding particle size and zeta potential. To study the bone targeting potential of these conjugates, an in vitro HAP binding assay was established. The obtained binding data indicate that CHOL-TOE-BP is useful as targeting device for liposomal drug delivery to bone.
The potential and limitations of targeted delivery of anticancer agents with colloidal particulat... more The potential and limitations of targeted delivery of anticancer agents with colloidal particulate carriers is the subject of this contribution. Because over the years liposomes have gained the most attention as carrier system in the category of colloidal carrier systems, this paper focuses on the utility of the liposomal system for tumor targeting. It is imperative that an intended therapeutic application of liposomes should be well matched with the liposome behavior in vivo. Therefore, the in vivo fate of the first-generation liposomes and the more recently developed second-generation liposomes (surface-modified liposomes such as the immunoli¬ posomes and long-circulating liposomes) is analyzed in terms of accessibility of target sites, time-, and sitecontrolled drug release and potential target sites for rational targeted delivery are discussed. A few examples of areas in cancer chemotherapy, with a strong rationale for the use of liposomes, are given. It is concluded that, although several options are available on the drawing board, issues such as tumor cell heterogeneity, ac¬ cess to the target site, shedding of antigens, and target site-specific release of the liposome-associated drug need to be addressed early in the development process.
Mesna is used as a mucolytic agent in CO, PD* and CF-patients and to prevent chemical cystitis in... more Mesna is used as a mucolytic agent in CO, PD* and CF-patients and to prevent chemical cystitis in cancer patients after nigh dose cyclophosphamide of ifosfamide. In urological patients with an" Indianapouch" or" Brickerbladder" mucus formadon is sometimes a ...
Macromolecular carrier materials based on N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic... more Macromolecular carrier materials based on N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic and well-characterized drug delivery systems that have been extensively evaluated in the past two decades, both at the preclinical and at the clinical level. Using several different imaging agents and techniques, HPMA copolymers have been shown to circulate for prolonged periods of time, and to accumulate in tumors both effectively and selectively by means of the Enhanced Permeability and Retention (EPR) effect. Because of this, HPMA-based macromolecular nanotheranostics, i.e. formulations containing both drug and imaging agents within a single formulation, have been shown to be highly effective in inducing tumor growth inhibition in animal models. In patients, however, as essentially all other tumor-targeted nanomedicines, they are generally only able to improve the therapeutic index of the attached active agent by lowering its toxicity, and they fail to improve the efficacy of the intervention. Bearing this in mind, we have recently reasoned that because of their biocompatibility and their beneficial biodistribution, nanomedicine formulations might be highly suitable systems for combination therapies. In the present manuscript, we briefly summarize several exemplary efforts undertaken in this regard in our labs in the past couple of years, and we show that long-circulating and passively tumor-targeted macromolecular nanotheranostics can be used to improve the efficacy of radiochemotherapy and of chemotherapy combinations.
To assist intravenously administered anticancer agents in achieving proper circulation times and ... more To assist intravenously administered anticancer agents in achieving proper circulation times and tumor concentrations, and to thereby improve the balance between their efficacy and their toxicity, a large number of drug delivery systems have been designed and evaluated over the years. Clinically relevant examples of such nanometer-sized carrier materials are liposomes, polymers, micelles and antibodies. In the vast majority of cases, however, and especially in patients, nanomedicine formulations are only able to attenuate the toxicity of the conjugated or entrapped chemotherapeutic drug, and they generally fail to improve the efficacy of the intervention. To overcome this shortcoming, and to broaden the clinical applicability of tumor-targeted nanomedicines, in the past 5 years we have developed several concepts for using N-(2-hydroxypropyl)methacrylamide (HPMA)-based polymer therapeutics to enhance the efficacy of combined modality anticancer therapy. Regarding surgery, HPMA copoly...
Second Congress of the European Society for Clinical Neuropharmacology Wtirzburg, November 9-11, ... more Second Congress of the European Society for Clinical Neuropharmacology Wtirzburg, November 9-11, 1995
A folate-poly(ethylene glycol) conjugate capable of covalent coupling to primary amines present a... more A folate-poly(ethylene glycol) conjugate capable of covalent coupling to primary amines present at the surface of polyplexes was developed. Coating of poly(dimethylaminomethyl methacrylate (pDMAEMA)-based polyplexes with this folate-pEG conjugate led to a sharp decrease of the z-potential, and a small increase in particle size. The size of the particles in isotonic medium did not change markedly in time demonstrating that rather stable particles were formed. The in vitro cellular toxicity of the pEGylated polyplexes with and without folate ligands was lowered considerably compared to uncoated polyplexes. The toxicity observed for the targeted pEGylated polyplexes was slightly higher than that of corresponding untargeted polyplexes, which might indicate an increased cellular association of targeted polyplexes. Transfection of OVCAR-3 cells in vitro was markedly increased compared to untargeted pEGylated polyplexes, suggesting targeted gene delivery.
A major limiting factor in the full exploitation of therapeutically active peptide and protein dr... more A major limiting factor in the full exploitation of therapeutically active peptide and protein drugs prepared synthetically and/ or by recombinant DNA technology, is the lack of availability of appropriate delivery systems. This contribution deals with the potential of liposomes for improved delivery of these drugs. Our current efforts in this field are briefly described: (1) longcirculating liposomes for prolonged systemic delivery of peptide drugs; (2) liposomes as carriers of interleukin-2 in locoregional anticancer immunotherapy; (3) plasminogen-bearing liposomes for the targeted delivery of tissue plasminogen activator; (4) immunoliposomes for the targeted delivery of diphtheria toxin; and (5) immunoliposomes bearing enzymes (immunoenzymosomes) for site-specific activation of prodrugs.
Understanding the role of the epidermal growth factor receptor (EGFR) in cellular signalling proc... more Understanding the role of the epidermal growth factor receptor (EGFR) in cellular signalling processes underlying malignancy has enabled the development of rationally designed EGFR-targeted therapeutics. Strategies have been devised to interfere with the EGFR signalling at three different levels: at the extracellular level, competing with ligand binding; at the intracellular level, inhibiting the activation of the tyrosine kinase; or at the mRNA level, modulating the expression of the EGFR protein. Each of these strategies has proven to have an antitumour effect mediated by events such as inhibition of cell proliferation, induction of apoptosis, decrease of cellular invasion and migration; and/or inhibition of angiogenesis. Furthermore, the combination of these strategies with traditional chemotherapy or radiotherapy has generally resulted in enhanced antitumour effects. Likewise, the benefit of interfering simultaneously with different signalling pathways has been documented to improve tumour growth inhibition. These preclinical results have encouraged clinical studies that led to the FDA approval of three drugs. However, finding the perfect strategy for each individual patient appears to be a limiting factor, demanding further research to be able to generate relevant molecular expression profiles on a case-to-case basis. Taken together, a successful EGFR inhibition will require a better understanding of signalling pathways in combination with the development of rationally designed effective molecules.
Objective. Angiogenic endothelial cells can function as phagocytes, and phagocytosis is initiated... more Objective. Angiogenic endothelial cells can function as phagocytes, and phagocytosis is initiated via the opsonin lactadherin. In this study, we examined the interaction between lactadherin-opsonized erythrocytes with reduced deformability and angiogenic endothelium, as loss of deformability is characteristic for suicidal and aged erythrocytes. Materials and Methods. We used the Arg-Gly-Asp (RGD)-modified erythrocyte model and investigated the deformability parameter by cross-linking erythrocyte membranes through treatment with glutaraldehyde. Association in vitro with primary endothelial cells was detected by flow cytometry and visualized by light, fluorescent, and electron microscopy. Involvement of two crucial factors in phagocytosis, a v-integrins and Rho guanosine triphosphatase family member Rac1, was studied using small interfering RNA technology. Modified erythrocytes were administered in vivo into tumor-bearing mice to detect phagocytosis by endothelial cells. Results. Glutaraldehyde-treated (rigid) RGD-modified erythrocytes showed a strongly enhanced endothelial cell association compared to flexible RGD-modified erythrocytes. Knockdown by small interfering RNA lipoplexes of a v-integrins and Rac1 confirmed classical tethering and internalization of rigid RGD-erythrocytes. Upon in vivo administration, tumor endothelium showed pronounced erythrophagocytosis. Conclusion. The pronounced phagocytosis of opsonized erythrocytes with reduced deformability by angiogenic growth factorLactivated endothelial cells evokes new insights in endothelial cell function and suggests a role for these endothelial cells in (hematological) disorders because of their capacity to clear disordered erythrocytes.
In the course of an inflammation, vascular endothelial cells (VECs) are strongly involved in proc... more In the course of an inflammation, vascular endothelial cells (VECs) are strongly involved in processes like leukocyte recruitment, cytokine production, and angiogenesis. Specific interference in these processes may yield great therapeutic benefit in the treatment of (chronic) inflammatory disorders. Drug targeting to VECs at inflamed sites may allow such intervention. VECs at inflamed sites represent a very well-accessible target cell population for circulating drug-targeting systems, which may also be selectively distinguished from normal VECs by the expression of several cell surface receptors involved in the inflammation. One group of specifically expressed molecules are the adhesion molecules (AMs), which have a major function in adhesion of cells to each other, to the extracellular matrix, or in the adhesion and subsequent recruitment of circulating immune cells. This review describes AMs with regard to their function in the inflammatory disease and their usefulness in functioning as a specific target receptor for drug-targeting approaches in general and with an emphasis on liposome-based drug delivery.
Reproduce material from Drug Discovery Today? This publication and the contributions it contains ... more Reproduce material from Drug Discovery Today? This publication and the contributions it contains are protected by the copyright of Elsevier. Except as outlined in the terms and conditions (see p. X), no part of this journal can be reproduced without written permission from Elsevier,
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