ABSTRACT
AIM: To investigate the modulatory effect of recombinant-expressed vasoactive intestinal peptide (VIP) analogue (rVIPa) on trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats. METHODS: Forty-eight rats were randomized into six groups: normal control group (Control), model control group (TNBS), ethanol treatment group (ETOH), and VIP treatment groups with different dosage (rVIPa1nmol, rVIPa2nmol, rVIPa4nmol). Diarrhea and bloody stool were observed. Colonic damage was evaluated histologically. The levels of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), myeloperoxidase (MPO) and endotoxin in colonic tissue and serum were determined by enzyme-linked immunosorbent assay (ELISA). The expression of occludin, ZO-1, Toll-like receptor 4 (TLR4), and nuclear factor-kappa B p65 (NF-κB p65), IκBα, and p-IκBα were detected by Western blot. RESULTS: Administration with 2 nmol rVIPa prevented TNBS-induced necrosis, hyperemia, swelling, inflammation, etc., pathologic changes observed in the inner surface of colon in experimental rats. Moreover, rVIPa significantly decreased colonic TNF-α level (P < 0.001), MPO activity (P < 0.001) and serum endotoxin level (P < 0.01), and remarkably increased colonic IL-10 content (P < 0.001) in rats with TNBS-induced colitis. Furthermore, compared to the TNBS-induced colitis group, 2 nmol rVIPa treatment up-regulated the levels of occludin (P < 0.05) and ZO-1 (P < 0.05), NF-κB p65 (P < 0.01) and IκBα (P < 0.001), and down-regulated the levels of TLR4. CONCLUSION: rVIPa ameliorates TNBS-induced colonic injury and inflammation and effectively protected the intestinal mucosal barrier function in rats. The mechanism may be related to TLR4/NF-κB-mediated signaling pathway. rVIPa could be used as a new alternative therapy for intestinal inflammatory disorders.
Subject(s)
Colon/drug effects , Crohn Disease/drug therapy , Gastrointestinal Agents/therapeutic use , Intestinal Mucosa/drug effects , Vasoactive Intestinal Peptide/analogs & derivatives , Animals , Colon/pathology , Crohn Disease/chemically induced , Crohn Disease/pathology , Diarrhea/chemically induced , Diarrhea/drug therapy , Diarrhea/pathology , Disease Models, Animal , Gastrointestinal Agents/pharmacology , Humans , Intestinal Mucosa/pathology , Male , NF-kappa B/metabolism , Necrosis/drug therapy , Necrosis/pathology , Rats , Rats, Sprague-Dawley , Recombinant Proteins/pharmacology , Recombinant Proteins/therapeutic use , Signal Transduction/drug effects , Toll-Like Receptor 4/metabolism , Treatment Outcome , Trinitrobenzenesulfonic Acid/toxicityABSTRACT
The aim of present study was to develop a respirable powder (RP) of a shortened vasoactive intestinal peptide (VIP) analog for inhalation. VIP and C-terminally truncated VIP analogs were synthesized with a solid-phase method. A structure-activity relationship (SAR) study was carried out in terms with binding and relaxant activities of the peptides. Prepared RP formulation of a shortened VIP analog was physicochemically characterized by morphological, in vitro aerodynamic, and pharmacological assessments. The SAR study demonstrated that the N-terminal 23 amino acid residues were required for biological activity of VIP. Upon chemical modification of VIP(1-23), [R15, 20, 21 , L17 ]-VIP(1-23) was newly developed, which had higher binding activity in rat lung and smooth muscle relaxant effect in mouse stomach than VIP(1-23). The [R15, 20, 21 , L17 ]-VIP(1-23)-based RP, [R15, 20, 21 , L17 ]-VIP(1-23)/RP, exhibited fine in vitro inhalation performance. Airway inflammation evoked by sensitization of antigen in rats was attenuated by pre-treatment with the [R15, 20, 21 , L17 ]-VIP(1-23)/RP at a dose of 50 µg-[R15, 20, 21 , L17 ]-VIP(1-23)/rat as evidenced by a 70% reduction of recruited inflammatory cells in bronchoalveolar lavage fluid. On the basis of these results, [R15, 20, 21 , L17 ]-VIP(1-23)/RP might be a promising agent for treatment of airway inflammatory diseases.
Subject(s)
Asthma/drug therapy , Gastrointestinal Agents/administration & dosage , Gastrointestinal Agents/chemical synthesis , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Animals , Bronchoalveolar Lavage Fluid/cytology , Disease Models, Animal , Gastrointestinal Agents/pharmacology , Lung/drug effects , Male , Mice , Powders , Rats , Stomach/drug effects , Structure-Activity Relationship , Vasoactive Intestinal Peptide/chemistryABSTRACT
Vasoactive intestinal peptide (VIP) is an anti-inflammatory neuropeptide recently identified as a potential antimicrobial peptide. To overcome the metabolic limitations of VIP, we modified the native peptide sequence and generated two stable synthetic analogues (VIP51 and VIP51(6-30)) with better antimicrobial profiles. Herein we investigate the effects of both VIP analogues on cell viability, membrane integrity, and ultrastructure of various bacterial strains and Leishmania species. We found that the two VIP derivatives kill various non-pathogenic and pathogenic Gram-positive and Gram-negative bacteria as well as the parasite Leishmania major through a mechanism that depends on the interaction with certain components of the microbial surface, the formation of pores, and the disruption of the surface membrane. The cytotoxicity of the VIP derivatives is specific for pathogens, because they do not affect the viability of mammalian cells. Docking simulations indicate that the chemical changes made in the analogues are critical to increase their antimicrobial activities. Consequently, we found that the native VIP is less potent as an antibacterial and fails as a leishmanicidal. Noteworthy from a therapeutic point of view is that treatment with both derivatives increases the survival and reduces bacterial load and inflammation in mice with polymicrobial sepsis. Moreover, treatment with VIP51(6-30) is very effective at reducing lesion size and parasite burden in a model of cutaneous leishmaniasis. These results indicate that the VIP analogues emerge as attractive alternatives for treating drug-resistant infectious diseases and provide key insights into a rational design of novel agents against these pathogens.
Subject(s)
Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Leishmania major/drug effects , Vasoactive Intestinal Peptide/pharmacology , Amino Acid Sequence , Animals , Endotoxemia/drug therapy , Endotoxemia/microbiology , Female , Gram-Negative Bacteria/genetics , Gram-Positive Bacteria/genetics , Hydrogen Bonding , Leishmania major/genetics , Leishmania major/ultrastructure , Leishmaniasis, Cutaneous/drug therapy , Leishmaniasis, Cutaneous/parasitology , Mice , Mice, Inbred BALB C , Microbial Viability/drug effects , Microscopy, Electron , Models, Molecular , Molecular Sequence Data , Mutation , Protein Conformation , Sepsis/drug therapy , Sepsis/microbiology , Survival Analysis , Treatment Outcome , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/chemistryABSTRACT
Recent years are the time of dynamic development of pulmonary arterial pressure pharmacotherapy. By introducing the goal oriented therapy the survival in this group of patients has significantly increased. Apart from the pharmacotherapy used according to the ESC guidelines, new attempts of interventional treatment based on denervation of pulmonary artery have also been taken. Constantly, the new clinical trials (tests?) of drugs acting via new metabolic pathways have been conducted. They include for example: soluble guanylate cyclase stimulators, tyrosine kinase inhibitors, serotonin receptors inhibitors, Rhokinase inhibitors, VIP analogues. One of the newmedicines is riociguat, the effectiveness and safety of which have been confirmed in the PATENT and CHEST study. However, the small number and clinical diversity in the group of the PAH patients cause significant difficulties with the extrapolation of the results of trials according to the guidelines of the therapy.
Subject(s)
Antihypertensive Agents/therapeutic use , Denervation/methods , Hypertension, Pulmonary/therapy , Pulmonary Artery/innervation , rho-Associated Kinases/antagonists & inhibitors , Guanylate Cyclase/therapeutic use , Humans , Protein-Tyrosine Kinases/antagonists & inhibitors , Pyrazoles/therapeutic use , Pyrimidines/therapeutic use , Serotonin Antagonists/therapeutic use , Vasoactive Intestinal Peptide/analogs & derivativesABSTRACT
In both mammals and birds, vasoactive intestinal polypeptide (VIP) neurons and fibers are present in virtually every brain area that is important for social behavior. VIP influences aggression in birds, social recognition in rodents, and prolactin secretion in both taxa, but other possible functions in social modulation remain little explored. VIP effects are mediated by VPAC receptors, which bind both VIP and pituitary adenylate cyclase activating peptide. Within the lateral septum and medial bed nucleus of the stria terminalis, VPAC receptors are found at higher densities in gregarious finch species relative to territorial species, suggesting that VPAC receptor activation promotes social contact and/or preference for larger groups. Here we here test this hypothesis in zebra finches (Taeniopygia guttata), and also examine the relevance of VPAC receptors to anxiety-like processes. Intraventricular infusions of the VPAC receptor antagonist, neurotensin6-11 mouseVIP7-28, strongly reduce social contact when animals are tested in a novel environment, and exert sex-specific effects on grouping behavior. Specifically, VPAC receptor antagonism reduces gregariousness in females but increases gregariousness in males. Interestingly, VPAC antagonism in the medial pallium (putative prefrontal cortex homologue) significantly reduces gregariousness in both sexes, suggesting site-specific effects of VIP signaling. However, VPAC antagonism does not modulate novel-familiar social preferences in a familiar environment or general anxiety-like behaviors. The current results suggest that endogenous activation of VPAC receptors promotes social contact under novel environmental conditions, a function that may be accentuated in gregarious species. Moreover, endogenous VIP modulates gregariousness in both males and females.
Subject(s)
Behavior, Animal/physiology , Exploratory Behavior/physiology , Finches/physiology , Neurotensin/pharmacology , Prefrontal Cortex/drug effects , Receptors, Vasoactive Intestinal Peptide/physiology , Social Behavior , Vasoactive Intestinal Peptide/pharmacology , Animals , Anxiety/physiopathology , Behavior, Animal/drug effects , Environment , Exploratory Behavior/drug effects , Female , Male , Prefrontal Cortex/physiology , Receptors, Vasoactive Intestinal Peptide/antagonists & inhibitors , Recombinant Fusion Proteins/pharmacology , Signal Transduction/drug effects , Signal Transduction/physiology , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/antagonists & inhibitorsABSTRACT
The present study aimed to design a PEGylated VIP derivative, [Arg(15, 20, 21), Leu(17)]-VIP-GRR (IK312532), with improved metabolic stability, and develop its respirable powder (RP) formulation for inhalation therapy. IK312532 was chemically conjugated with PEG (5 kDa, P5K), the physicochemical and biochemical properties of which were characterized by CD spectral analysis, binding assays, and metabolic stability. CD spectral analysis demonstrated that PEG conjugation had no impact on the conformational structure of IK312532. Although the receptor-binding activity of IK312532/P5K (IC50: 82 nM) was estimated to be ca. 30-fold less than that of IK312532 (IC50: 2.8 nM), the metabolic stability of IK312532/P5K was highly improved. The IK312532/P5K was jet-milled and blended with lactose carrier particles to provide RP formulation of IK312532/P5K (IK312532/P5K-RP). In vitro inhalation performance and in vivo pharmacological effects of the IK312532/P5K-RP in antigen-sensitized rats were also evaluated. In cascade impactor analyses, fine particle fraction of IK312532/P5K-RP was calculated to be ca. 37%. Insufflation of IK312532/P5K-RP (150 µg of IK312532/P5K) in antigen-sensitized rats resulted in marked attenuation of inflammatory events, as evidenced by significant decreases in inflammatory biomarkers and granulocyte recruitment in pulmonary tissue 24h after the antigen challenge. From these findings, PEGylation of a VIP derivative, as well as its strategic application to the RP formulation, may be a viable approach to improve its therapeutic potential for the treatment of airway inflammatory diseases.
Subject(s)
Anti-Inflammatory Agents/chemistry , Polyethylene Glycols/chemistry , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Allergens , Animals , Anti-Inflammatory Agents/administration & dosage , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/cytology , Cell Count , L-Lactate Dehydrogenase/metabolism , Lung/drug effects , Lung/metabolism , Lung/pathology , Male , Ovalbumin , Particle Size , Peroxidase/metabolism , Pneumonia/drug therapy , Pneumonia/metabolism , Polyethylene Glycols/administration & dosage , Rats , Rats, Sprague-Dawley , Vasoactive Intestinal Peptide/administration & dosage , Vasoactive Intestinal Peptide/chemistryABSTRACT
The present study was undertaken to develop a respirable sustained-release powder (RP) formulation of long-acting VIP derivative, [Arg(15, 20, 21), Leu(17)]-VIP-GRR (IK312532), using PLGA nanospheres (NS) with the aim of improving the duration of action. NS formulation of IK312532 (IK312532/NS) was prepared by an emulsion solvent diffusion method in oil, and a mixture of the IK312532/NS and erythritol was jet-milled and mixed with lactose carrier to obtain the IK312532/NS-RP. Physicochemical properties were characterized focusing on appearance, particle size, and drug release, and in vivo pharmacological effects were assessed in antigen-sensitized rats. The IK312532/NS with a diameter of 140 nm showed a biphasic release pattern in distilled water with ca. 20% initial burst for 30 min and a sustained slow release up to ca. 55% for 24h. Laser diffraction analysis demonstrated that IK312532/NS-RP had fine dispersibility and suitable particle size for inhalation. In antigen-sensitized rats, insufflated IK312532/NS-RP (10 µg of IK312532/rat) could suppress increases of granulocyte recruitment and myeloperoxidase in pulmonary tissue for up to 24h after antigen challenge, although IK312532-RP at the same dose was less effective with limited duration of action. From these findings, newly prepared IK312532/NS-RP might be of clinical importance in improving duration of action and medication compliance for treatment of airway inflammatory diseases.
Subject(s)
Asthma/drug therapy , Pneumonia/drug therapy , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Animals , Bronchoalveolar Lavage Fluid/cytology , Delayed-Action Preparations , Disease Models, Animal , Granulocytes/drug effects , Granulocytes/physiology , Lung/drug effects , Lung Diseases/drug therapy , Male , Nanospheres , Peroxidase/metabolism , Rats , Rats, Sprague-Dawley , Respiratory System/drug effects , Respiratory System/immunology , Vasoactive Intestinal Peptide/administration & dosage , Vasoactive Intestinal Peptide/pharmacology , Vasoactive Intestinal Peptide/therapeutic useABSTRACT
Vasoactive intestinal peptide (VIP) attenuates experimental acute pancreatitis (AP) by inhibition of cytokine production from inflammatory cells. It has been suggested that reactive oxygen species (ROS) as well as cytokines play pivotal roles in the early pathophysiology of AP. This study aimed to clarify the effect of VIP on the oxidative condition in pancreas, especially pancreatic acinar cells (acini). Hydrogen peroxide (H(2)O(2))-induced intracellular ROS, assessed with CM-H(2)DCFDA, increased time- and dose-dependently in acini isolated from rats. Cell viability due to ROS-induced cellular damage, evaluated by MTS assay, was decreased with ≥100 µmol/L H(2)O(2). VIP significantly inhibited ROS production from acini and increased cell viability in a dose-dependent manner. Expression of antioxidants including catalase, glutathione reductase, superoxide dismutase (SOD) 1 and glutathione peroxidase was not altered by VIP except for SOD2. Furthermore, Nox1 and Nox2, major components of NADPH oxidase, were expressed in pancreatic acini, and significantly increased after H(2)O(2) treatment. Also, NADPH oxidase activity was provoked by H(2)O(2). VIP decreased NADPH oxidase activity, which was abolished by PKA inhibitor H89. These results suggested that VIP affected the mechanism of ROS production including NADPH oxidase through induction of a cAMP/PKA pathway. In conclusion, VIP reduces oxidative stress in acini through the inhibition of NADPH oxidase. These results combined with findings of our previous study suggest that VIP exerts its protective effect in pancreatic damage, not only through an inhibition of cytokine production, but also through a reduction of the injury caused by oxidative stress.
Subject(s)
Acinar Cells/drug effects , Acinar Cells/metabolism , NADPH Oxidases/antagonists & inhibitors , Oxidative Stress/drug effects , Pancreas, Exocrine/cytology , Vasoactive Intestinal Peptide/metabolism , Vasoactive Intestinal Peptide/pharmacology , Acinar Cells/cytology , Animals , Cells, Cultured , Male , NADPH Oxidases/metabolism , Rats , Rats, Inbred Lew , Reactive Oxygen Species/metabolism , Vasoactive Intestinal Peptide/analogs & derivativesABSTRACT
Excitotoxicity is a key molecular mechanism of perinatal brain damage and is associated with cerebral palsy and long term cognitive deficits. VIP induces a potent neuroprotection against perinatal excitotoxic white matter damage. VIP does not prevent the initial appearance of white matter lesion but promotes a secondary repair with axonal regrowth. This plasticity mechanism involves an atypical VPAC2 receptor and BDNF production. Stable VIP agonists mimic VIP effects when given systemically and exhibit a large therapeutic window. Unraveling cellular and molecular targets of VIP effects against perinatal white matter lesions could provide a more general rationale to understand the neuroprotection of the developing white matter against excitotoxic insults.
Subject(s)
Brain/embryology , Cerebral Palsy/metabolism , Neuroprotective Agents/therapeutic use , Vasoactive Intestinal Peptide/physiology , Vasoactive Intestinal Peptide/therapeutic use , Animals , Brain/metabolism , Cerebral Palsy/prevention & control , Humans , Neuronal Plasticity/drug effects , Receptors, Vasoactive Intestinal Peptide, Type II/metabolism , Receptors, Vasoactive Intestinal Peptide, Type II/physiology , Signal Transduction/drug effects , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/metabolismABSTRACT
Vasoactive intestinal peptide (VIP) has been considered as a promising drug candidate for asthma and COPD because of its potent immunomodulating and anti-inflammatory activities. Recently, our group developed a new VIP derivative, [R(15, 20, 21), L(17), A(24,25), des-N(28)]-VIP-GRR (IK312548), with improved chemical and metabolic stability. In the present study, a dry powder inhaler system of IK312548 was designed for inhalation therapy with minimal systemic side effects, the physicochemical properties of which were also evaluated with a focus on morphology, particle size distribution, inhalation performance, and peptide stability. Laser diffraction and cascade impactor analysis suggested high dispersion and deposition in the respiratory organs with a fine particle fraction of 31.2%. According to UPLC/ESI-MS and circular dichroic spectral analyses, no significant changes in the purity and structure of VIP derivative were observed during preparation of respirable formulation. Anti-inflammatory properties of IK312548 respirable powder (RP) were characterized in antigen-sensitized asthma/COPD-model rats. There were marked inflammatory cells infiltrated into the lung tissues of experimental asthma/COPD-model rats; however, intratracheal administration of IK312548-RP led to significant reductions of recruited inflammatory cells in lung tissues and BALF by 72 and 78%, respectively. Thus, respirable powder formulation of IK312548 might be a promising medication for asthma, COPD, and other airway inflammatory diseases.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Asthma/drug therapy , Pulmonary Disease, Chronic Obstructive/drug therapy , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/pharmacokinetics , Asthma/physiopathology , Disease Models, Animal , Drug Stability , Dry Powder Inhalers , Inflammation/drug therapy , Inflammation/physiopathology , Lung/drug effects , Lung/physiopathology , Male , Particle Size , Pulmonary Disease, Chronic Obstructive/physiopathology , Rats , Rats, Sprague-Dawley , Tissue Distribution , Vasoactive Intestinal Peptide/administration & dosage , Vasoactive Intestinal Peptide/pharmacokinetics , Vasoactive Intestinal Peptide/pharmacologyABSTRACT
Vasoactive intestinal peptide (VIP) has been thought to be a promising candidate for asthma/chronic obstructive pulmonary disease (COPD), and our group previously developed several long-lasting VIP derivatives. The objective of the present study was to clarify the therapeutic potential of new VIP derivatives with improved chemical and metabolic stability. Exposure of rat alveolar L2 cells to cigarette smoke extract (CSE) for 1h led to release of lactate dehydrogenase (LDH) and decreased viability in a CSE concentration-dependent manner. There appeared to be marked induction of apoptosis after CSE exposure, as demonstrated by 59% elevation of caspase-3 activity and TUNEL staining. In contrast, a stabilized VIP derivative, [R(15,20,21), L(17)]-VIP-GRR (IK312532), at a concentration of 10(-7)M, exhibited 71% attenuation of LDH release and 85% decrease of the number of apoptotic cells. In addition to IK312532, new VIP derivatives also showed anti-apoptotic effects against CSE toxicity and marked reduction of nitric oxide production. In terms of cytoprotective effects, [R(15,20,21), L(17), A(24,25), des-N(28)]-VIP-GRR was more effective than VIP and IK312532, possibly due to the improved stability. Thus, the present study is the first to demonstrate that novel stabilized VIP derivatives exert anti-apoptotic and cytoprotective effects on CSE-induced cytotoxicity.
Subject(s)
Apoptosis/drug effects , Cytotoxins/pharmacology , Nicotiana/chemistry , Pulmonary Alveoli/cytology , Smoke/adverse effects , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/pharmacology , Amino Acid Substitution , Animals , Caspase 3/metabolism , Caspase 7/metabolism , Cell Line , Cell Survival/drug effects , Cytoprotection , L-Lactate Dehydrogenase/metabolism , Nitric Oxide/metabolism , Protein Stability , Rats , Nicotiana/toxicity , Vasoactive Intestinal Peptide/chemistryABSTRACT
Vasoactive intestinal peptide (VIP) is a pleiotropic neuropeptide with potent anti-inflammatory properties, and its receptor, VPAC1, mediates most of the anti-inflammatory effects of VIP. Diabetes mellitus is characterized by increased oxidation and inflammation due to persistent hyperglycemia. This research was performed to investigate the effects of VIP and a VPAC1 agonist on streptozotocin (STZ)-induced type 1 diabetic mice. Intraperitoneal injection of VIP and VPAC1 agonist (50nmol/kg/day in saline) over a 28-day period (1) decreased food intake, (2) increased body weight, (3) improved visceral index, (4) increased the fasting plasma insulin levels, (5) decreased the fasting plasma glucose, (6) improved the glucose tolerance, (7) decreased pancreas H(2)O(2) and malondialdehyde (MDA) and (8) increased total antioxidant activity (T-AOC) in the liver, spleen and pancreas. The results of histopathological and immunohistochemical analysis showed that VIP and the VPAC1 agonist improved the structure and cellularity of islets and ameliorated the insulin-secreting activity of islets. Additionally, administration of VIP or the VPAC1 agonist not only significantly decreased the plasma TNFα and CRP and promoted IL-10 in diabetic mice but also blocked the increased NF-κB activity of pancreatic tissue in diabetic mice. Furthermore, the VPAC1 agonist displayed stronger effects than VIP. These results show that both VIP and VPAC1 agonist ameliorated STZ-induced diabetes and protected mice against oxidative stress and inflammation associated diabetes, with VPAC1 being the receptor most responsible for these positive effects in diabetic mice.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Antioxidants/pharmacology , Diabetes Mellitus, Experimental/drug therapy , Hypoglycemic Agents/pharmacology , Peptide Fragments/pharmacology , Receptors, Vasoactive Intestinal Polypeptide, Type I/agonists , Vasoactive Intestinal Peptide/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Antioxidants/metabolism , Antioxidants/therapeutic use , Blood Glucose/drug effects , Blood Glucose/metabolism , Body Weight/drug effects , C-Reactive Protein/metabolism , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Experimental/pathology , Eating/drug effects , Glucose Tolerance Test , Hydrogen Peroxide/metabolism , Hypoglycemic Agents/therapeutic use , Insulin/blood , Insulin/metabolism , Interleukin-10/blood , Liver/drug effects , Liver/metabolism , Male , Malondialdehyde/metabolism , Mice , Mice, Inbred Strains , NF-kappa B/metabolism , Pancreas/drug effects , Pancreas/metabolism , Pancreas/pathology , Peptide Fragments/therapeutic use , Spleen/drug effects , Spleen/metabolism , Tumor Necrosis Factor-alpha/blood , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/therapeutic useABSTRACT
OBJECTIVE: We pharmacologically characterized pituitary adenylate cyclase-activating polypeptides (PACAPs), vasoactive intestinal peptide (VIP) and the VPAC(1), VPAC(2) and PAC(1) receptors in human meningeal (for their role in migraine) and coronary (for potential side effects) arteries. METHODS: Concentration response curves to PACAP38, PACAP27, VIP and the VPAC(1) receptor agonist ([Lys15,Arg16,Leu27]-VIP[1-7]-GRF[8-27]) were constructed in the absence or presence of the PAC(1) receptor antagonist PACAP6-38 or the VPAC(1) receptor antagonist, PG97269. mRNA expression was measured using qPCR. RESULTS: PACAP38 was less potent than VIP in both arteries. Both peptides had lower potency and efficacy in meningeal than in coronary arteries, while mRNA expression of VPAC(1) receptor was more pronounced in meningeal arteries. PACAP6-38 reduced the E(max) of PACAP27, while PG97269 right-shifted the VIP-induced relaxation curve only in the coronary arteries. CONCLUSION: The direct vasodilatory effect of VIP and PACAP might be less relevant than the central effect of this compound in migraine pathogenesis.
Subject(s)
Coronary Vessels/physiology , Meningeal Arteries/physiology , Migraine Disorders/physiopathology , Pituitary Adenylate Cyclase-Activating Polypeptide , Receptors, Vasoactive Intestinal Peptide , Adult , Aged , Coronary Vessels/drug effects , Female , Gene Expression/drug effects , Gene Expression/physiology , Humans , Male , Meningeal Arteries/drug effects , Middle Aged , Migraine Disorders/drug therapy , Peptide Fragments/pharmacology , Pituitary Adenylate Cyclase-Activating Polypeptide/adverse effects , Pituitary Adenylate Cyclase-Activating Polypeptide/antagonists & inhibitors , Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , Pituitary Adenylate Cyclase-Activating Polypeptide/pharmacology , RNA, Messenger/metabolism , Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide/agonists , Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide/antagonists & inhibitors , Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , Receptors, Vasoactive Intestinal Peptide/agonists , Receptors, Vasoactive Intestinal Peptide/antagonists & inhibitors , Receptors, Vasoactive Intestinal Peptide/genetics , Receptors, Vasoactive Intestinal Peptide, Type II/agonists , Receptors, Vasoactive Intestinal Peptide, Type II/antagonists & inhibitors , Receptors, Vasoactive Intestinal Peptide, Type II/genetics , Receptors, Vasoactive Intestinal Polypeptide, Type I/agonists , Receptors, Vasoactive Intestinal Polypeptide, Type I/antagonists & inhibitors , Receptors, Vasoactive Intestinal Polypeptide, Type I/genetics , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/pharmacology , Vasodilation/drug effects , Vasodilation/physiology , Vasodilator Agents/pharmacologyABSTRACT
Cigarette smoke (CS) has been identified as a predominant causative factor for chronic obstructive pulmonary disease (COPD), so CS-exposed rodent model of COPD has drawn considerable interest and attention for fundamental study and drug discovery. In the present study, using experimental COPD model rats, the therapeutic potential of a newly prepared respirable powder (RP) formulation of a long-acting VIP derivative, [Arg(15,20,21), Leu(17)]-VIP-GRR (IK312532), was assessed with a focus on pro-inflammatory biomarkers, morphological and histochemical changes, and infiltrated cells in the respiratory system. CS exposure of rats for 11 days led to the marked infiltration of inflammatory cells, except for eosinophils, in bronchiolar epithelium, followed by goblet cell metaplasia and hyperplasia. However, inhalation of IK312532-RP (50µg/rat) in the CS-exposed rats resulted in 74 and 71% reductions of granulocyte recruitment in bronchoalveolar lavage fluids and lung tissues, respectively, with 68% decrease of goblet cells. Biomarker study demonstrated that the inhaled IK312532-RP could suppress the CS-evoked increase of myeloperoxidase in both plasma and lung by 87 and 70%, respectively, possibly leading to potent suppression of neutrophilic inflammatory symptoms. The results from TUNEL staining were indicative of apoptotic damage in respiratory tissues of the CS-exposed rats, and there appeared to be marked decrease of TUNEL-positive cells in the CS-exposed rat with inhaled IK312532-RP. The present findings suggest that an inhalable formulation of IK312532 might be efficacious as a therapy for COPD or other airway inflammatory diseases because of its potent immunomodulating activities.
Subject(s)
Inflammation/drug therapy , Neutrophils/physiology , Nicotiana , Smoke/adverse effects , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Biomarkers , Eosinophil Peroxidase/metabolism , Inflammation/chemically induced , Lung Diseases/chemically induced , Lung Diseases/drug therapy , Lung Diseases/pathology , Male , Neutrophils/drug effects , Peroxidase/metabolism , Powders , Rats , Rats, Sprague-Dawley , Respiratory Mucosa/drug effects , Respiratory Mucosa/pathology , Vasoactive Intestinal Peptide/administration & dosage , Vasoactive Intestinal Peptide/chemistry , Vasoactive Intestinal Peptide/therapeutic useABSTRACT
Glioblastoma multiforme (GBM) is the most aggressive form of brain tumor in adults. This cancer has an infiltrative nature and the median survival of patients is about one year. Vasoactive intestinal peptide (VIP) belongs to a structurally related family of polypeptides and is a major regulatory factor in the central and peripheral nervous systems. VIP regulates proliferation of astrocytes and of numerous cancer cell lines and modulates migration in prostatic and colonic cancer cell lines. Little is known about the involvement of VIP and its receptors (VIP-receptor system) in proliferation or migration of GBM cells. The effects of VIP, PACAP and of synthetic VIP antagonists were tested in two human GBM cell lines, M059K and M059J, established from two different parts of a single tumor. In these cells, the data revealed that the VIP-receptor system did not affect proliferation but controlled cell migration. Indeed, in M059K cells which express components of the VIP receptor system, the VIP receptor antagonists and a PACAP antibody enhanced migration. The VIP receptor antagonists increased generation of typical migration-associated processes: filopodia and lamellipodia, and activation of Rac1 and Cdc42 GTPases. Reciprocally, in M059J cells which poorly express the VIP-receptor system, treatments with the agonists VIP and PACAP resulted in decreased cell migration. Furthermore, the peptides appeared to act through a subclass of binding sites displaying an uncommon very high affinity for these ligands. Taken together, these observations suggest that components of the VIP-receptor system negatively regulate cell migration, thus showing potential anti-oncogenic properties.
Subject(s)
Cell Movement/physiology , Receptors, Vasoactive Intestinal Peptide/metabolism , Vasoactive Intestinal Peptide/pharmacology , Blotting, Western , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Cyclic AMP/metabolism , Humans , Immunohistochemistry , Polymerase Chain Reaction , Receptors, Vasoactive Intestinal Peptide/agonists , Receptors, Vasoactive Intestinal Peptide/antagonists & inhibitors , Vasoactive Intestinal Peptide/analogs & derivativesABSTRACT
We previously demonstrated that the gamma-glutamyl 16 amine derivative of vasoactive intestinal peptide (VIP) acts as structural VIP agonist with affinity and potency higher than VIP. Herein, we have evaluated the effects of VIP and gamma-Gln16-diaminopropane derivative of VIP (VIP-DAP3) on the proliferation and protection from oxidative stress induced by hydrogen peroxide (H2O2) on epidermoid carcinoma cell lines. We have found that 10(-11) M VIP-DAP3 completely antagonized the inhibition induced by H2O2 on both cell proliferation and S-phase distribution while these effects were only partially antagonized by equimolar concentrations of VIP. Moreover, both oxidative stress and intracellular lipid oxidation induced by H2O2 were reduced by VIP and completely antagonized by VIP-DAP3. Thereafter, we have found that H2O2 increased p38 kinase activity and both HSP70 and HSP27 expression. VIP and VIP-DAP3 again antagonized these effects partially or totally, respectively. H2O2 reduced the activity of extracellular signal-regulated kinases Erk-1/2 and Akt, signalling proteins involved in proliferation/survival pathways. Again VIP restored the activity of both kinases while VIP-DAP3 caused indeed an increase of their activity as compared to untreated cells. These data suggest that VIP-DAP3 has a stronger anti-oxidative activity as compared to VIP likely based on its super-agonistic binding on the putative receptor.
Subject(s)
Antioxidants/pharmacology , Carcinoma, Squamous Cell/drug therapy , Oropharyngeal Neoplasms/drug therapy , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/pharmacology , Carcinoma, Squamous Cell/metabolism , Cell Line, Tumor , Humans , MAP Kinase Signaling System/drug effects , Mitogen-Activated Protein Kinases/metabolism , Oropharyngeal Neoplasms/metabolism , Oxidative Stress/drug effectsABSTRACT
Vasoactive intestinal peptide (VIP) exerts immunomodulating and anti-inflammatory activities through its specific receptors, such as VPAC1 and 2 receptors. Previously, a stabilized VIP derivative, [R(15,20,21), L(17)]-VIP-GRR (IK312532), was proposed as a candidate of anti-asthma drug, and a dry powder inhaler system of IK312532 was also developed for inhalation therapy with minimal systemic side-effects. In the present study, the anti-inflammatory properties of IK312532 respirable powder (RP) were characterized in an asthma/COPD-like animal model, with the use of newly developed ovalbumin (OVA)-RP for lung inflammation. Marked inflammatory events in the lung were observed after OVA-RP challenge in rats as evidenced by significant increase of inflammatory biomarkers such as eosinophil peroxidase (EPO), myeloperoxidase (MPO) and lactate dehydrogenase (LDH). However, intratracheal administration of IK312532-RP led to significant attenuation of plasma EPO, MPO and LDH activities, as well as significant reduction of recruited inflammatory cells in BALF, especially macrophages and eosinophils. In the rats pretreated with IK312532-RP, histochemical examinations revealed that the inflammatory cells infiltrating to the lung and the epithelial wall thickness decreased significantly by 85% and 58%, respectively. Thus, inhalable powder formulation of IK312532 exerts its anti-inflammatory activity by suppressing granulocyte recruitment to the lung and epithelial hyperplasia, followed by the reduction of cytotoxic peroxidases.
Subject(s)
Anti-Inflammatory Agents , Asthma/drug therapy , Powders/therapeutic use , Pulmonary Disease, Chronic Obstructive/drug therapy , Vasoactive Intestinal Peptide/analogs & derivatives , Administration, Inhalation , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/therapeutic use , Asthma/immunology , Biomarkers/metabolism , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/immunology , Disease Models, Animal , L-Lactate Dehydrogenase/metabolism , Male , Peroxidase/metabolism , Pulmonary Disease, Chronic Obstructive/immunology , Rats , Rats, Sprague-Dawley , Vasoactive Intestinal Peptide/administration & dosage , Vasoactive Intestinal Peptide/therapeutic useABSTRACT
Previously, [R(15,20,21), L(17)]-VIP-GRR (IK312532), a long-acting VIP derivative, was proposed as potential drug candidate for the treatment of asthma/COPD. The present work is aimed to elucidate solution-state stability of IK312532 and to develop further stabilized derivative with equipotent or higher biological functions. A stability study on IK312532 was carried out in solution state, and degradation mechanism was deduced by UPLC-MS and amino acid analyses. Three novel VIP derivatives were designed and chemically synthesized on the basis of stability data, being subjected to physicochemical and pharmacological characterization. Solution-state stability studies revealed the gradual degradation of IK312532, following pseudo-first-order kinetics. Chemical modification of IK312532, mainly position at 24, resulted in marked improvement of stability, although the chemical modification had no influence on the secondary structure, receptor binding, and activation of adenylate cyclase in rat lung cells. Novel derivatives also exhibited more potent neurite outgrowth in rat pheochromocytoma PC12 cells when compared to VIP and IK312532, possibly due to improved stability. Deamination of Asn at position 24 might be responsible for degradation of VIP derivative, and stability and chemical modification studies led us to the successful development of novel VIP derivatives with higher stability and biological functions.
Subject(s)
Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/chemistry , Amino Acid Sequence , Animals , Cells, Cultured , Chemical Phenomena , Molecular Sequence Data , Neurites/drug effects , Neurites/physiology , PC12 Cells , Protein Stability , Rats , Vasoactive Intestinal Peptide/genetics , Vasoactive Intestinal Peptide/pharmacologyABSTRACT
Vasoactive intestinal peptide (VIP) is an interesting example of a 28-amino acid neuropeptide that is abundantly expressed in discrete brain regions/neurons and hence may contribute to brain function. This short review summarizes my own point of view and encompasses 25 years of work and over 100 publications targeting the understanding of VIP production and biological activity. The review starts with our original cloning of the VIP gene, it then continues to discoveries of regulation of VIP synthesis and the establishment of the first VIP transgenic mice. The review ends with the identification of novel VIP analogs that helped decipher VIP's important role during development, in regulation of the biological clock(s) and diurnal rhythms, sexual activity, learning and memory as well as social behavior, and cancer. This review cites only articles that I have coauthored and gives my own perspective of this exciting ever-growing field.
Subject(s)
Research , Vasoactive Intestinal Peptide , Amino Acid Sequence , Animals , Biological Clocks/physiology , Brain/physiology , Cell Proliferation , Cyclic AMP/metabolism , Cytokines/metabolism , History, 20th Century , History, 21st Century , Humans , Intercellular Signaling Peptides and Proteins/metabolism , Molecular Sequence Data , Neoplasms/metabolism , RNA, Messenger/metabolism , Research/history , Sequence Alignment , Synapses/physiology , Vasoactive Intestinal Peptide/analogs & derivatives , Vasoactive Intestinal Peptide/genetics , Vasoactive Intestinal Peptide/metabolismABSTRACT
A novel peptide combination consisting of four synthetic neuropeptide analogs of Vasoactive Intestinal Peptide (VIP), Bombesin, Substance P and Somatostatin has been found to have potent anticancer activity in vitro and in vivo. The receptors of these four neuropeptides are known to be over expressed in various cancers. We have found the presence of native neuropeptides in the culture supernatant of the primary tumor cells of human colon adenocarcinomas. It was further demonstrated by receptor-ligand assays that not only do these tumor cells synthesize and secrete four peptide hormones but also possess specific high affinity receptors on their surface. Screening a large panel of analogs to the four peptide hormones on tumor cell proliferation led to the identification of four cytotoxic analogs, the combination of which was code-named DRF7295. The design and synthesis of the peptide analogs have been described in this paper. In vitro anticancer activity of DRF7295 was studied in a large panel of human tumor cells. Gastrointestinal tumor cells of the colon, pancreas and duodenum were found to be most sensitive to DRF7295 with moderate activity seen in glioblastoma, prostate, leukemia and those of oral cancer cells. Efficacy studies in xenograft models of colon and duodenum resulted in T/C% of less than 40%, which is indicative of strong tumor regressing potential of DRF7295 in gastrointestinal cancers. Acute and long-term toxicity studies as well as safety pharmacology studies conducted indicate the safety of the drug upon systemic administration with no significant adverse pharmacological effects.