Azelastine Research Articles

ＴＮＢＳ大腸炎モデルにおける塩酸Ａｚｅｌａｓｔｉｎｅの効果およびその作用機序に関する免疫組織化学的検討

クローン病の実験モデルである2,4,6-trinitrobenzene sulfonic acid(TNBS)大腸炎を作成し,塩酸Azelastine(AZ)を投与して病変粘膜局所における免疫担当細胞の動態を免疫組織化学的に検討した.TNBS大腸炎群の潰瘍形成部ではICAM-1,Mac-1陽性の単核球の強い浸潤があり,隣接した粘膜固有層にはICAM-1陽性の血管内皮細胞とLFA-1,Mac-1陽性の単核球の増加がみられた.AZ投与群では,潰瘍形成を示したのは5例中1例のみであった.肉眼的に炎症のない粘膜においては,TNBS大腸炎群に比べICAM-1の発現とLFA-1,Mac-1陽性細胞は軽度に認められるのみであった.IgG,IgE含有細胞および好酸球数も有意に低下していた(p<0.05).AZはこれら接着分子の発現の抑制やIgG,IgE産生の抑制を介し病変の軽減をもたらすことが示唆された.

Suppression by azelastine hydrochloride of NF-kappa B activation involved in generation of cytokines and nitric oxide.

The influence of the anti-allergy agent azelastine hydrochloride (Azeptin) on NF-kappa B activation associated with the generation of cytokines and nitric oxide (NO) was investigated in various kinds of human and mouse cells. Azeptin dose-dependently suppressed both DNA and protein synthesis in human gingival fibroblasts (HF) and also suppressed blastogenesis of human peripheral blood lymphocytes (PBL). Generation of tumor necrosis factor-alpha, interleukin 1-beta, granulocyte-macrophage colony-stimulating factor and interleukin-6 from 10(-5) M Azeptin-treated PBL and human monocytes (HM) was decreased to approximately 1/3 to 2/3 of the control levels. In parallel with the decreased cytokine generation, each cytokine mRNA was less expressed in the presence of 10(-5) M Azeptin. In addition, both inducible nitric oxide synthase-mRNA level and NO generation in mouse peritoneal macrophages were suppressed by 10(-5) M Azeptin. Being compatible with those results, Azeptin (10(-5) M) suppressed activation of NF-kappa B in PBL, HM and HF. These results appear to indicate that suppression of cytokine and NO generation by Azeptin results at least partially from the inhibition of NF-kappa B activation.

Suppression by Azelastine Hydrochloride of NF-κB Activation Involved in Generation of Cytokines and Nitric Oxide

The influence of the anti-allergy agent azelastine hydrochloride (Azeptin®) on NF-κB activation associated with the generation of cytokines and nitric oxide (NO) was investigated in various kinds of human and mouse cells. Azeptin dose-dependently suppressed both DNA and protein synthesis in human gingival fibroblasts (HF) and also suppressed blastogenesis of human peripheral blood lymphocytes (PBL). Generation of tumor necrosis factor-alpha, interleukin 1-beta, granulocyte-macrophage colony-stimulating factor and interleukin-6 from 10−5 M Azeptin-treated PBL and human monocytes (HM) was decreased to approximately 1/3 to 2/3 of the control levels. In parallel with the decreased cytokine generation, each cytokine mRNA was less expressed in the presence of 10−5 M Azeptin. In addition, both inducible nitric oxide synthase-mRNA level and NO generation in mouse peritoneal macrophages were suppressed by 10−5 M Azeptin. Being compatible with those results, Azeptin (10−5 M) suppressed activation of NF-κB in PBL, HM and HF. These results appear to indicate that suppression of cytokine and NO generation by Azeptin results at least partially from the inhibition of NF-κB activation.

Efficacy and Tolerability of Azelastine Nasal Spray in the Treatment of Allergic Rhinitis: Large Scale Experience in Community Practice

SummaryTwo Spanish prospective monitoring studies evaluated efficacy and tolerability of azelastine nasal spray* containing azelastine hydrochloride for allergic rhinitis. Both studies were conducted by community practitioners over two weeks (Study I) or one month (Study II).The numbers of patients recruited were 3680 (I) and 4002 (II). Of these, 56.1% (I) and 51.7% (II) had been previously treated with oral antihistamines with/without other medications. Patients rated the severity of 10 symptoms of allergic rhinitis as absent, mild. moderate or severe. Azelastine nasal spray was generally administered at a dose of one spray puff (0.14 mg) per nostril twice daily. Follow-up was after 14 days (I) or 31 days (II), when symptoms were rated and patients questioned about treatment.Assessment was by a sum score for all 10 symptoms. A symptom sum score of 1620 occurred in 21.1% (I) and 13.7% (II) of patients before treatment and only 0.8% (I) and 0.6% (II) after treatment. A symptom sum score of 11—15 occurred in 35.9% (I) and 30.5% (II) of patients before treatment and only 2.6% (I) and 2.8% (II) after treatment. Overall, 92.3% (I) and 90.7% (II) of patients were completely pee of adverse events, 7.0% (I) and 8.8% (II) experienced one and 0.7% (I) and 0.6% (II) two adverse events. The number of doctors who rated efficacy as either very good or good was 89.4% (I) and 84.6% (II). General tolerance was rated as good or very good by 97.5% (I) and 97.3% (II), and local tolerance by 93.1% (I) and 91.5% (II) of physicians, respectively.Overall, azelastine nasal spray was highly effective and very well tolerated in normal clinical practice.

Azelastine hydrochloride inhibits platelet activating factor-like activity in human eosinophils

We investigated the inhibitory effect of azelastine hydrochloride (azelastine), an anti-asthmatic drug, on platelet-activating factor (PAF)-like activity in eosinophils obtained from asthmatic and non-asthmatic patients. Eosinophils were preincubated with or without azelastine and stimulated with f-Met-Leu-Phe (fMLP, 10 μmol) for 15 min. PAF-like activity was detected by aggregation of washed guinea-pig platelets. PAF-like activity released from asthmatic eosinophils without preincubation of azelastine was 2.36 [1.02] (mean [SD], ng/10 7 cells) in supernatants and 13.87 [4.77] in cell pellets. After preincubation with 10 −8, 10 −6, and 10 −4 M azelastine, PAF-like activity reduced to 1.85 [0.46] (mean [SD], ng/10 7 cells), 1.11 [0.14], and 0.88 [0.09] ( n = 15) in the supernatants, and 11.83 [2.93], 8.32 [1.41], and 6.27 [1.25] ( n = 15) in the cell pellets, respectively. PAF-like activity in non-asthmatic eosinophils without preincubation of azelastine was 2.01 [0.86] (mean [SD], ng/10 7 cells) in supernatants and 7.44 [0.99] in cell pellets. After preincubation with 10 −8, 10 −6, and 10 −4 M azelastine, PAF-like activity reduced to 1.73 [0.64] (mean [SD], ng/10 7 cells), 1.12 [0.23], and 0.84 [0.17] ( n = 20) in the supernatants, and 6.26 [2.08], 4.65 [0.88], and 3.02 [0.43] ( n = 20) in the cell pellets, respectively. Our results showed that preincubation with azelastine caused a dose-dependent inhibition of intra and extracellular PAF-like activity from asthmatic and non-asthmatic eosinophils in the same manner.

Inhibition of collagen expression by azelastine hydrochloride in cultured skin fibroblasts from normal individuals and scleroderma patients

Inhibition of collagen expression by azelastine hydrochloride in cultured skin fibroblasts from normal individuals and scleroderma patients

Inhibition of collagen expression by azelastine hydrochloride in cultured skin fibroblasts from normal individuals and scleroderma patients.

The effects of azelastine hydrochloride on cell proliferation and collagen synthesis in cultured human skin fibroblasts were studied. Azelastine inhibited cell proliferation during proliferating cell phases. Azelastine was found to inhibit collagen synthesis without altering cell proliferation during quiescent phases. It did not alter the ratio of type I to III collagen synthesis. Northern blot analysis of collagen chain mRNAs revealed that the levels of alpha1 (I), alpha1 (III) and alpha1 (VI) mRNAs were reduced by azelastine treatment, whereas the level of alpha2 (VI), alpha3 (VI) mRNAs were not significantly changed. These results suggest that azelastine modulates collagen synthesis at a pretranslational level. Azelastine inhibited collagen synthesis in fibroblasts from scleroderma patients to the same extent as in normal skin fibroblasts. This drug may be useful in the treatment of fibrotic diseases.

塩酸アゼラスチンの咳しきい値に及ぼす影響

塩酸アゼラスチンの咳しきい値に及ぼす影響

Azelastine and flezelastine as reversing agents of multidrug resistance: Pharmacological and molecular studies

The effects of two new phthalazinone derivatives, azelastine (AZ) and flezelastine (FZ), on the reversal of resistance to doxorubicin (dox) were studied using two variants of the rat C6 glioblastoma cell line, selected with dox (C6 0.5) or with vincristine (C6 1V). Both lines presented a multidrug-resistant phenotype which was, in the case of C6 0.5 cells, likely to be accompanied by an additional mechanism leading to intracellular tolerance of the drug. Both AZ and FZ reversed dox resistance in a concentration-dependent manner, and FZ was shown to be at least three times more potent than AZ. FZ was able, at a relatively high concentration (30 μM), to completely restore dox sensitivity in both cell lines. Both drugs were able to virtually restore dox accumulation to the level reached in sensitive cells, and, interestingly, this complete restoration occurred at lower concentrations of modulator than required for complete reversal of resistance. FZ was able to reverse dox intracellular tolerance of C6 0.5 cells and to restore dox accumulation at the ic 50 to the level observed in sensitive cells. AZ and FZ both inhibited azidopine binding to membrane preparations of C6 0.5 and C6 1V cells, although FZ was more potent. Both drugs more successfully inhibited azidopine binding to membranes prepared from C6 1V cells (which express the mdr1b gene product) than to membranes from C6 0.5 cells (which express the mdr1a gene product). In view of its potent activity on MDR, further preclinical evaluation of FZ is warranted.

Contrasting influence of peplomycin and azelastine hydrochloride (Azeptin) on reactive oxygen generation in polymorphonuclear leukocytes, cytokine generation in lymphocytes, and collagen synthesis in fibroblasts.

The influence of peplomycin (PLM) and azelastine hydrochloride (Azeptin) on reactive oxygen (RO) and cytokine generation was examined in human peripheral blood mononuclear leukocytes, polymorphonuclear leukocytes (PMN), and rabbit alveolar macrophages (RAM). In addition, the influence of these drugs on DNA and collagen synthesis was investigated in human gingival and rabbit pulmonary fibroblasts. In vitro, PLM increased the FMLP- and PMA-induced chemiluminescence and superoxide (O2-) generation in human PMN and RAM in a dose-dependent manner. In contrast to PLM, Azeptin dose-dependently suppressed RO generation. Such contrasting actions of PLM and Azeptin were also observed in RAM and PMN obtained from rabbits treated with PLM or Azeptin. Even when human PMN were preincubated with 10-100 micrograms/ml of PLM, the increase in RO generation was negligible in the presence of 10(-5) M Azeptin in the culture medium. No increases in RO generation were observed in RAM or PMN obtained from rabbits that had received PLM (0.1 mg/kg per day) and Azeptin (0.04 mg/kg per day) concomitantly. PLM suppressed superoxide dismutase activity in RAM and human PMN, while Azeptin did not affect this activity. In vitro, PLM up-regulated the release of interleukin-1 beta, interleukin-6, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor both from human cells and from RAM and pulmonary fibroblasts. In the generation of these cytokines, Azeptin abrogated the up-regulatory action of PLM. PLM and Azeptin also had contrasting actions in [3H]thymidine and [3H]proline incorporation in human and rabbit fibroblasts. Furthermore, protein tyrosine phosphorylation, in particular that of a 115-kDa protein in human PMN, was suppressed by Azeptin and enhanced by PLM. These results seem to indicate that up-regulated RO and collagen generation are the causative factors of PLM-induced pulmonary fibrosis and that Azeptin may suppress the adverse effect.

Effect of azelastine on endotoxin-induced airway hyperresponsiveness in mice.

We examined the role of lipopolysaccharide (LPS) on the pulmonary inflammatory process in mice, including the release of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) by macrophages, and investigated the mechanism of action of azelastine (AZ) on the release of these two cytokines. Intratracheal instillation of 1.0 microgram/ml LPS into BALB/c mice caused a significant increase in both IL-1 beta and TNF-alpha in aqueous lung extracts. These changes were modified, under control conditions, by a single dose of 0.05 mg/kg of AZ administered 1 and 11 h after LPS infusion. Intratracheal instillation of LPS also caused a significant increase in bronchial hyperresponsiveness to methacholine (Mch). AZ significantly inhibited Mch responsiveness in LPS-infused mice compared with nontreated control mice. Our results suggested that intratracheal instillation of LPS induces the secretion of macrophage cytokines in the airways of mice, accounting, at least partly, for LPS-induced airway hyperresponsiveness. Our results also indicate that the attenuating effect of AZ on LPS-induced airway hyperresponsiveness could be explained by its inhibitory effect on macrophage cytokine production.

Role of thromboxane A 2 synthetase inhibitors in the treatment of patients with bronchial asthma

Asthma results from complex interactions among inflammatory cells, mediators, and the cells and tissues resident in the airways and is characterized by airway obstruction, airway inflammation, and airway hyperresponsiveness. Treatment of asthma should address not only the airway obstruction but also the chronic inflammation that may lead to hyperresponsiveness. In Japan, where the death rate from asthma has not increased despite increasing numbers of patients, treatment of bronchial asthma relies on the use of oral prophylactic antiasthma drugs, such as thromboxane synthetase inhibitors. Thromboxanes may facilitate the effect of acetylcholine on the airways and may be involved in hyperresponsiveness. Experiments using animal models have shown that thromboxane synthetase inhibitors have prevented increased airway reactivity after exposure to allergens and irritants. Double-blind clinical trials have shown that treatment with the thromboxane A 2 synthetase inhibitor ozagrel hydrochloride significantly reduced the need for concomitant steroid therapy, compared with treatment with azelastine hydrochloride. This review discusses the role of thromboxane A 2 synthetase inhibitors in the treatment of patients with bronchial asthma.

じん麻疹，湿疹・皮膚炎に対する塩酸アゼラスチンの臨床的検討 特に他剤無効例に対する塩酸アゼラスチンの有用性の検討

じん麻疹，湿疹・皮膚炎に対する塩酸アゼラスチンの臨床的検討 特に他剤無効例に対する塩酸アゼラスチンの有用性の検討

塩酸アゼラスチンの熱傷進展に対する効果

熱傷の急性期における塩酸アゼラスチン(アゼプチン®)の熱傷の進展に対する効果を動物モデルを用いて検討した。背部にsuperficial dermal burn(SDB)を受傷させたラットにアゼプチン®を1mg/kg静脈内投与し, 組織内のLTB4濃度を測定した。その結果, 塩酸アゼラスチン投与群においては組織内LTB4濃度の有意な低下が認められ, 組織学的にも炎症細胞の浸潤の抑制が認められた。このことにより塩酸アゼラスチンは熱傷急性期の炎症を抑制し, 熱傷の進展防止に有用であると考えられた。

アゼプチンの鼻アレルギー鼻閉に対する効果

アゼプチンの鼻アレルギー鼻閉に対する効果

Contrasting influence of peplomycin and azelastine hydrochloride (Azeptin) on reactive oxygen generation in polymorphonuclear leukocytes, cytokine generation in lymphocytes, and collagen synthesis in fibroblasts

Contrasting influence of peplomycin and azelastine hydrochloride (Azeptin) on reactive oxygen generation in polymorphonuclear leukocytes, cytokine generation in lymphocytes, and collagen synthesis in fibroblasts

Suppression of respiratory burst of polymorphonuclear leukocytes by azelastine hydrochloride (Azeptin).

The inhibitory action of azelastine hydrochloride (Azeptin) on the respiratory burst in peripheral polymorphonuclear leukocytes (PMN) and pulmonary alveolar macrophages (PAM) has been studied. Azeptin in vitro suppressed chemiluminescence and superoxide (O2-) generation by human PMN in a dose- and time-dependent manner. Phorbol myristyl acetate (PMA) and formyl-methionyl-leucyl-phenylalanine (FMLP)-induced O2- generation were strongly suppressed by 10(-6) M and 10(-5) M Azeptin, respectively. PMN and PAM from rabbits injected with Azeptin 0.2 mg.kg-1 for 5 days showed lower chemiluminescence and O2- generation than cells from untreated rabbits. Nitroblue tetrazolium reduction activity in human PMN was suppressed by treatment of PMN with 10(-6) M Azeptin for 6 h. Inositol trisphosphate, intracellular free calcium, and protein kinase C activity were decreased by 10(-6) M to 10(-5) M Azeptin. The tyrosine phosphorylation of many proteins, especially a 115 kDa protein, was suppressed by 10(-5) M Azeptin. However, superoxide dismutase activity in PMN, PAM, and lung tissue samples was only slightly decreased, even when the rabbits were treated with 1.0 mg.kg-1 Azeptin for 5 days. The results suggest that Azeptin suppresses multiple signal transduction steps in the respiratory burst of PMN. This suppressive action should be very useful in the prevention and treatment of reactive oxygen-associated disorders.

Immunosuppressive Effects of Azelastine Hydrochloride on Contact Hypersensitivity and T-Cell Proliferative Response: A Comparative Study with FK-506

Azelastine hydrochloride (AZE) is an anti-allergic drug that inhibits the release of various chemical mediators from mast cells. We compared the immunosuppressive effects of AZE and FK-506 in vivo and in vitro. Topical application of AZE strongly inhibited the efferent phase of contact hypersensitivity, as did application of FK-506. In in vitro experiments, we found that 1) the suppression by AZE on interleukin (IL)-2 production from splenic T cells was partial and considerably large amounts of IL-2 were still produced, even in the presence of 10(-5) M of AZE, which was in sharp contrast to the observed marked inhibition of [3H]-TdR incorporation; 2) AZE significantly inhibited the phorbol myristate acetate-induced IL-2 responsiveness; 3) AZE did not inhibit the IL-2 receptor alpha expression of activated T cells; and 4) the significant inhibitory action was still observed even when AZE was added at 48 h after the initiation of culture. In regard to FK-506, we found that 1) FK-506 completely blocked the production of IL-2; 2) exogeneous IL-2 consistently restored the FK-506-induced inhibition; 3) FK-506 affected the phorbol myristate acetate-induced IL-2 responsiveness very little, if any; and 4) the significant suppression was observed only when FK-506 was added within 24 h after the initiation of culture. Thus, AZE exerts its in vitro immunosuppressive activity preferentially by interfering with the IL-2 responsiveness, with partial inhibition of IL-2 production. Conversely, FK-506 acts as a strong inhibitor of IL-2 production without a prominent effect on IL-2 responsiveness. The immunosuppressive activity of AZE shown in vitro may also be operative in vivo and may be applicable for topical use.

抗アレルギー薬の新しい概念 基礎的実験結果から

According to the previous concepts of antiallergic drugs, it was difficult to select the essential antiallergic drugs which was necessary for treatment of patients with allergy. We, therefore, attempted to discover that antiallergic drugs have the any pharmacologic actions on the process of allergic response. We divided into three stages on the process of allergic response in this animal experiment. Antiallergic drugs used in this study were azelastine hydrochloride, ketotifen fumarate, tranilast and oxatomide. First, we observed the suppression of IgE production in rats on the administration of azelastine hydrochloride. Secondly, we observed that azelastine hydrochloride and ketotifen fumarate inhibited the interaction between anti-DNP-As and DNP-As on ELISA. Thirdly, we observed that tranilast and azelastine hydrochloride reduced, more effectively than any other antiallergic drug, the release of histamine from rabbit washed platelets by PAF.In addition, it was observed that azelastine hydrochloride and ketotifen inhibited dose-dependently the increase of permeability induced by PAF or histamine in rats. From these results, we could propose the new concepts that some of antiallergic drug inhibited the IgE production.

The effects of Roxithromycin and Azelastine hydrochloride in treatment of smell disturbance

The effects of Roxithromycin and Azelastine hydrochloride in treatment of smell disturbance