RESUMO
Myeloid-derived suppressor cells (MDSCs) are notorious for their pathological characteristics of immunosuppression and their promoting effect on cancers. They can induce the formation of pre-metastatic niche (PMN) characterized by inflammation, immunosuppression and vascular leakage, and promote pulmonary metastasis of breast cancer. Herein, a tumor targeting c(RGDfk) peptide modified low molecular-weight-heparin-all-trans-retinoic-acid (LMWH-ATRA) micellar nanoparticle loaded with chemotherapeutic drug doxorubicin (DOX) and immune adjuvant α-galactosylceramide (αGC) (RLA/DOX/αGC NP) was developed. The hydrophilic segment LMWH inhibited the recruitment of MDSCs by competitively binding with P-selectin on the surface of vascular endothelial cells (VECs), while the hydrophobic segment ATRA promoted the depletion of MDSCs by inducing their differentiation. Through the modulation of MDSCs, micelles can significantly improve the inflammatory and immunosuppressive microenvironment of the lung and tumor sites, and inhibit the formation of PMN. Not only this, the micelles also produced a synergistic effect with αGC, which effectively improved the anti-tumor immunity of tumor bearing mice and provided a promising therapeutic strategy for breast cancer and pulmonary metastasis.
Assuntos
Neoplasias Pulmonares , Células Supressoras Mieloides , Nanopartículas , Animais , Camundongos , Micelas , Heparina de Baixo Peso Molecular/química , Heparina de Baixo Peso Molecular/metabolismo , Heparina de Baixo Peso Molecular/farmacologia , Células Endoteliais , Neoplasias Pulmonares/patologia , Doxorrubicina/uso terapêutico , Tretinoína , Microambiente TumoralRESUMO
Phototherapy holds promise in cancer treatment for its prominent antitumor efficacy and low systematic toxicity compared with traditional chemotherapy. However, the higher risk of tumor metastasis caused by the severe hypoxic state during phototherapy is a threat in practical use. Here, in order to tackle this challenge, we developed a delivery system via loading the photosensitizer indocyanine green (ICG) into the low molecular weight heparin (LMWH) modified liposomes (LMWH-ICG-Lip) to realize the synergistic effects between photosensitizer and drug vehicle, achieving better phototherapeutic efficacy and meanwhile alleviating the potential risk of tumor metastasis caused by phototherapy. In this system, besides elongating the photosensitizers' circulation time and enhancing their accumulating efficacy to tumor tissues, LMWH itself also exhibited anti-metastasis efficacy via inhibiting adhesion of platelets to tumor cells and decreasing migration and invasion capability of tumor cells. In vivo efficacy evaluation was conducted on orthotopic 4T1 breast cancer model, and the system of LMWH-ICG-Lip could alleviate metastasis potential of residual tumor cells after irradiation, and elicit optimistic antitumor and anti-metastasis efficacy for phototherapy.
Assuntos
Neoplasias da Mama/terapia , Heparina de Baixo Peso Molecular/química , Verde de Indocianina/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Animais , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Feminino , Verde de Indocianina/química , Verde de Indocianina/farmacologia , Lipossomos , Camundongos , Metástase Neoplásica , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Based on the complementary effects of doxorubicin (DOX), all-trans retinoic acid (ATRA) and low molecular weight heparin (LMWH), the combination therapy of DOX, ATRA and LMWH was expected to exert the enhanced anti-tumor effects and reduce the side effects. In this study, amphiphilic LMWH-ATRA conjugate was synthesized for encapsulating the DOX. In this way, DOX, ATRA and LMWH were assembled into a single nano-system by both chemical and physical modes to obtain a novel anti-tumor targeting drug delivery system that can realize the simultaneous delivery of multiple drugs with different properties to the tumor. LMWH-ATRA nanoparticles exhibited good loading capacities for DOX with excellent physico-chemical properties, good biocompatibility, and good differentiation-inducing activity and antiangiogenic activity. The drug-loading capacity was up to 18.7% with an entrapment efficiency of 78.8%. It was also found that DOX-loaded LMWH-ATRA nanoparticles (DHR nanoparticles) could be efficiently taken up by tumor cells via endocytic pathway, and mainly distributed in cytoplasm at first, then transferred into cell nucleus. Cell viability assays suggested that DHR nanoparticles maintained the cytotoxicity effect of DOX on MCF-7 cells. Moreover, the in vivo imaging analysis indicated that DiR-loaded LMWH-ATRA nanoparticles could target the tumor more effectively as compared to free DiR. Furthermore, DHR nanoparticles possessed much higher anticancer activity and reduced side effects compared to free drugs solution. These results suggested that DHR nanoparticles could be considered as a promising targeted delivery system for combination cancer chemotherapy with lower adverse effects.
Assuntos
Antineoplásicos/uso terapêutico , Doxorrubicina/uso terapêutico , Portadores de Fármacos/uso terapêutico , Heparina de Baixo Peso Molecular/uso terapêutico , Nanopartículas/química , Tretinoína/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Doxorrubicina/química , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacologia , Quimioterapia Combinada , Heparina de Baixo Peso Molecular/química , Heparina de Baixo Peso Molecular/farmacologia , Humanos , Células MCF-7 , Camundongos Endogâmicos C57BL , Nanopartículas/toxicidade , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/patologia , Tretinoína/química , Tretinoína/farmacologiaRESUMO
It is well known that enoxaparin, a widely used anticoagulant and low-molecular-weight heparin containing a large number of oligosaccharides, possesses anti-inflammatory activity. Whilst enoxaparin has shown promising results in various inflammatory disorders, some of its oligosaccharides have anti-inflammatory properties and others increase the risk of bleeding due to their anticoagulant effects. The aim of this study was to develop an effective ion exchange chromatographic (IC) technique which allows the separation, isolation and, consequently, the identification of different oligosaccharides of enoxaparin with or without anticoagulant activity. The developed method utilises a semi-preparative CarboPac PA100 (9 × 250 mm) ion exchange column with sodium chloride gradient elution and UV detection at 232 nm. The method successfully resolved enoxaparin into more than 30 different peaks. IC-derived oligosaccharides with high, moderate, low or no anticoagulant activity were identified using an anti-factor Xa assay. The anti-inflammatory activity of selected oligosaccharides was investigated using the Griess assay. Using this technique, the oligosaccharides of enoxaparin with low or no anticoagulant activity, whilst exhibiting significant anti-inflammatory activity, could be fractionated. This technique can provide a platform to identify the oligosaccharides which are devoid of significant anticoagulant activity and are responsible for the therapeutic effects of enoxaparin that have been observed in various inflammatory conditions.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Anticoagulantes/farmacologia , Cromatografia por Troca Iônica/métodos , Enoxaparina/química , Heparina de Baixo Peso Molecular/química , Oligossacarídeos/isolamento & purificação , Animais , Sequência de Carboidratos , Linhagem Celular , Cromatografia por Troca Iônica/instrumentação , Avaliação Pré-Clínica de Medicamentos/métodos , Enoxaparina/farmacologia , Desenho de Equipamento , Inibidores do Fator Xa , Heparina de Baixo Peso Molecular/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Dados de Sequência Molecular , Peso Molecular , Oligossacarídeos/farmacologia , Reprodutibilidade dos TestesRESUMO
The regulation of angiogenesis is an interesting area to consider for novel therapeutic approaches to rheumatoid arthritis (RA). Chemically modified heparins have been developed as possible candidates for angiogenesis inhibitor; however, they have a major clinical drawback in exhibiting poor oral bioavailability. Here, orally absorbable O-desulfated low molecular weight heparin (ODS-LMWH) derivatives were newly synthesized by conjugating 2-O- or 6-O-desulfated LMWH with deoxycholic acid (DOCA) or bisDOCA (a dimer of DOCA), and their physicochemical properties, antiangiogenic potency and pharmacokinetic profiles were assessed. After selecting the best candidate among those derivatives, its therapeutic efficacy on arthritis was investigated in a murine collagen antibody-induced arthritis (CAIA) model. ODS-LMWH derivatives significantly inhibited the capillary-like tube formation of human umbilical vein endothelial cells (HUVECs) and basic fibroblast growth factor (bFGF)-induced angiogenesis in the Matrigel plug assay. Among all the compounds, 6ODS-LHbD showed the highest oral bioavailability in rats (19.3%). In the CAIA mouse model, 6ODS-LHbD (10 mg/kg, p.o., S.I.D.) significantly inhibited neovascularization in the joint, the increase of hind-paw thickness, and the structural damage in the bone. Therefore, 6ODS-LHbD would be a promising candidate for an orally active drug for the treatment of RA.
Assuntos
Inibidores da Angiogênese/administração & dosagem , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Ácido Desoxicólico/administração & dosagem , Heparina de Baixo Peso Molecular/administração & dosagem , Administração Oral , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacocinética , Animais , Artrite Experimental/patologia , Artrite Reumatoide/patologia , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/patologia , Ácido Desoxicólico/análogos & derivados , Ácido Desoxicólico/química , Ácido Desoxicólico/farmacocinética , Heparina de Baixo Peso Molecular/química , Heparina de Baixo Peso Molecular/farmacocinética , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Sprague-Dawley , Enxofre/químicaRESUMO
This study was designed to develop a solid oral dosage form of deoxycholic acid (DOCA)-conjugated low-molecular-weight heparin (LMWH) and to evaluate its oral absorption, distribution, and metabolic stability for the prospect of providing an orally bioavailable LMWH. The LMWH derivative (LHD) was synthesised and then formulated with solubilisers and other pharmaceutical excipients to form a solid tablet. Its absorption and distribution after oral administration were evaluated in mice, rats, and monkeys. The in vitro metabolic stability of LHD was examined by liver microsome assays. More than 80% of LHD was released from the tablet within 60 minutes, guaranteeing rapid tablet disintegration after oral administration. Oral bioavailability of LHD in mice, rats and monkeys were 16.1 ± 3.0, 15.6 ± 6.1, and 15.8 ± 2.5%, respectively. After the oral administration of 131I-tyramine-LHD, most of the absorbed drug remained in the blood circulation and was eliminated mainly through the kidneys. LHD was hardly metabolised by the liver microsomes and showed a stable metabolic pattern similar to that of LMWH. In a rat thrombosis model, 10 mg/kg of orally administered LHD reduced thrombus formation by 60.8%, which was comparable to the anti-thrombotic effect of the subcutaneously injected LMWH (100 IU/kg). Solid tablets of LHD exhibited high oral absorption and statistically significant therapeutic effects in preventing venous thromboembolism. Accordingly, LHD tablets are expected to satisfy the unmet medical need for an oral heparin-based anticoagulant as an alternative to injectable heparin and oral warfarin.
Assuntos
Anticoagulantes/farmacocinética , Ácido Desoxicólico/farmacocinética , Heparina de Baixo Peso Molecular/farmacocinética , Administração Oral , Animais , Anticoagulantes/administração & dosagem , Anticoagulantes/química , Disponibilidade Biológica , Ácido Desoxicólico/administração & dosagem , Ácido Desoxicólico/química , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Estabilidade de Medicamentos , Haplorrinos , Heparina de Baixo Peso Molecular/administração & dosagem , Heparina de Baixo Peso Molecular/química , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Engenharia de Proteínas , Ratos , Ratos Sprague-Dawley , Comprimidos , Trombose Venosa/prevenção & controleRESUMO
CB-01-05-MMX (LMW Heparin MMX), being developed by Cosmo Pharmaceuticals SpA, is a novel oral parnaparin sodium formulation for the potential treatment of ulcerative colitis. At the time of publication, clinical trial data were limited and the mechanism of action had not been elucidated. However, in one phase I and one phase IIb trial in patients with left-sided ulcerative colitis, CB-01-05-MMX had an acceptable safety profile and was not associated with bleeding complications, which is a known side effect of unfractionated heparin and low molecular weight heparin compounds previously evaluated for the treatment of ulcerative colitis. Evaluable efficacy data from the phase IIb trial revealed significant improvements in the clinical activity index values in the CB-01-05-MMX group, but this was not accompanied by mucosal healing. More research, including placebo-controlled trials in patients with left-sided ulcerative colitis and trials in patients with pancolitis, would determine the role of CB-01-05-MMX in the current spectrum of available medications for this condition.
Assuntos
Colite Ulcerativa/tratamento farmacológico , Portadores de Fármacos/química , Heparina de Baixo Peso Molecular/uso terapêutico , Administração Oral , Ensaios Clínicos como Assunto , Preparações de Ação Retardada , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Heparina de Baixo Peso Molecular/administração & dosagem , Heparina de Baixo Peso Molecular/antagonistas & inibidores , Heparina de Baixo Peso Molecular/química , Heparina de Baixo Peso Molecular/farmacocinética , Humanos , Resultado do TratamentoRESUMO
The initial choice of anticoagulant therapy administered in emergency departments for acute coronary syndromes (ACS) has important consequences for subsequent patient care, as neither unfractionated heparin (UFH) nor low-molecular-weight heparin (LMWH) are ideally suited for all potential clinical treatment pathways. UFH remains widely used for surgical interventions because of the ability to rapidly reverse its anticoagulant activity. However, the unpredictable pharmacokinetic profile of UFH presents safety issues, and the low subcutaneous bioavailability limits the utility of UFH for patients who are medically managed. LMWH has superior pharmacokinetic properties, but its anticoagulant activity cannot be effectively monitored or reversed during surgery. There is an unmet medical need for a baseline anticoagulant therapy that addresses these shortcomings while retaining the beneficial properties of both UFH and LMWH. We describe here M118, a novel LMWH designed specifically for use in the treatment of ACS. M118 shows broad anticoagulant activity, including potent activity against both factor Xa (~240 IU/mg) and thrombin (factor IIa; ~170 IU/mg), low polydispersity, high (78%) subcutaneous bioavailability in rabbits, and predictable subcutaneous and intravenous pharmacokinetics. Additionally, the anticoagulant activity of M118 is monitorable by standard coagulation assays and is reversible with protamine. M118 demonstrates superior activity to conventional LMWH in a rabbit model of abdominal arterial thrombosis without increasing bleeding risk, and is currently being evaluated in a phase II clinical trial evaluating efficacy and safety in patients undergoing percutaneous coronary intervention.
Assuntos
Síndrome Coronariana Aguda/tratamento farmacológico , Anticoagulantes/uso terapêutico , Doenças da Aorta/tratamento farmacológico , Arteriopatias Oclusivas/tratamento farmacológico , Animais , Anticoagulantes/administração & dosagem , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Anticoagulantes/toxicidade , Aorta Abdominal , Disponibilidade Biológica , Modelos Animais de Doenças , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Inibidores do Fator Xa , Hemorragia/induzido quimicamente , Heparina de Baixo Peso Molecular/administração & dosagem , Heparina de Baixo Peso Molecular/química , Heparina de Baixo Peso Molecular/isolamento & purificação , Heparina de Baixo Peso Molecular/uso terapêutico , Heparina de Baixo Peso Molecular/toxicidade , Injeções Intravenosas , Injeções Subcutâneas , Mucosa Intestinal/química , Masculino , Coelhos , Suínos , Trombina/antagonistas & inibidoresRESUMO
Heparins represent an efficient treatment of acute thrombosis and obstetric complications in antiphospholipid syndrome (APS). Enhanced microvesiculation of cell membranes, as detected by reduced membrane adhesion, can contribute to hypercoagulability in APS. Healthy donor IgG antibodies significantly increased beta2-glycoprotein I (beta2-GPI)-induced membrane adhesion, indicating that IgG antibodies might supplement the role of beta2-GPI in the regulation of membrane microvesiculation in healthy individuals. Anti-beta2-GPI IgG antibodies significantly reduced beta2-GPI-induced membrane adhesion, suggesting a direct role of anti-beta2-GPI antibodies in enhancing membrane microvesiculation in APS. Therapeutic concentration of nadroparin completely restored beta2-GPI-induced membrane adhesion in the presence of anti-beta2-GPI IgG antibodies. A novel anticoagulant mechanism of nadroparin in APS is suggested that supplements its direct effect on the coagulation cascade. Restoration of adhesion between negatively charged membranes in the presence of nadroparin might decrease shedding of microvesicles into the surrounding solution and could thus contribute to the efficacy of heparin treatment in APS.
Assuntos
Síndrome Antifosfolipídica/imunologia , Heparina de Baixo Peso Molecular/química , Lipídeos de Membrana/química , Fosfolipídeos/química , Adsorção/efeitos dos fármacos , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacologia , Anticoagulantes/química , Anticoagulantes/farmacologia , Síndrome Antifosfolipídica/sangue , Autoanticorpos/imunologia , Autoanticorpos/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Relação Dose-Resposta a Droga , Heparina de Baixo Peso Molecular/farmacologia , Humanos , Imunoglobulina G/imunologia , Imunoglobulina G/farmacologia , Fusão de Membrana/efeitos dos fármacos , Nadroparina/química , Nadroparina/farmacologia , Fosfatidilserinas/química , beta 2-Glicoproteína I/química , beta 2-Glicoproteína I/imunologia , beta 2-Glicoproteína I/farmacologiaRESUMO
The aim of this study was to investigate the influence of the proteolytic enzyme bromelain on the permeation of heparin across the gastrointestinal epithelial barrier. Stability of the complex and effect of heparin on the enzymatic activity of bromelain was analysed photometrically by measuring bromelain enzymatic activity in complex with the heparin. In vitro permeation studies were performed with Caco-2 cell monolayer and rat small intestinal mucosa in Ussing-type chambers, respectively. Results revealed that enzymatic activity of bromelain remained uninfluenced by the immobilization of heparin on it. Transport studies across Caco-2 cell monolayer and rat small intestine showed that the permeation of heparin could be significantly increased in presence of bromelain. In the study with Caco-2 cells, the most effective molar ratio of bromelain to heparin was 2:1, leading to 6.7-fold improvement in uptake, whereas the molar ratio 1:1 showed the highest permeation enhancing effect in the study on intestinal mucosa. This study provides evidence that heparin and bromelain form stable complexes leading to a significantly improved uptake of heparin.