Delay in ICR/f rat lens opacification by the instillation of eye drops containing disulfiram and hydroxypropyl-beta-cyclodextrin inclusion complex

In this study, we attempted to enhance disulfiram (DSF) solubility using a 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD) and hydroxypropylmethylcellulose (HPMC). We also investigated the effect of an HPbetaCD solution containing DSF and HPMC (DSF eye drops) on cataract development in ICR/f rat. The solubility of DSF increased with increasing HPbetaCD concentration, and the solubility of DSF in HPbetaCD solution containing 0.1% HPMC was approximately 20% greater than that of DSF in HPbetaCD solution without HPMC. In in vivo transcorneal penetration experiments using rabbits, only diethyldithiocarbamate (DDC) was detected (DSF was not detected) in the aqueous humor. This DSF-DDC conversion in the cornea was inhibited by treatment with a sulfhydryl (SH) inhibitor, p-mercuribenzoate and N-ethylmaleimide, in in vitro transcorneal penetration experiments using rabbit corneas. On the other hand, the instillation of 0.25% and 0.5% DSF eye drops delayed cataract development in ICR/f rats, a recessive-type hereditary cataractous strain. The present study demonstrates that DSF in HPbetaCD solution with HPMC is converted to DDC by the catalysis of proteins containing SH residues in the cornea, and this DDC may cause the delay in cataract development in ICR/f rats.     

Effect of disulfiram eye drops on lipid peroxide formation via excessive nitric oxide in lenses of hereditary cataract ICR/f rats

The ICR/f rat is a recessive-type hereditary cataractous strain, and opacity in the lens usually becomes evident at around 75 d of age. We previously found that the instillation of eye drops containing a disulfiram and hydroxypropyl-beta-cyclodextrin inclusion complex (DSF eye drops) delays lens opacification in ICR/f rats. In this study, we attempted to clarify the mechanisms of the delaying effect of DSF eye drops on cataract development in ICR/f rats. The calcium ion (Ca2+) content in the lenses of ICR/f rats increases at 77 d of age, and this elevation is preceded by a decrease in Ca2+-ATPase activity. On the other hand, the levels of nitric oxide (NO) and lipid peroxide (LPO) also increase in the lenses of ICR/f rats at 63 d of age, while the lenses are still transparent. The instillation of DSF eye drops reduces the changes in Ca2+ content, Ca2+-ATPase activity, NO and LPO levels in the lenses of ICR/f rats. The present study demonstrates that excessive NO production induces the increase in LPO, which causes the decrease in Ca2+-ATPase activity, and the increase in Ca2+ content in the lenses of ICR/f rat during cataract development. DSF eye drops have the ability to attenuate the increase in the NO and LPO levels, resulting in a delay in cataract development.     

Contribution of aldehyde dehydrogenase 3A1 to disulfiram penetration through monolayers consisting of cultured human corneal epithelial cells

We previously prepared 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD) solutions containing disulfiram (DSF) and hydroxypropylmethylcellulose (HPMC, DSF solutions), and found the instillation of this DSF solutions delayed lens opacification in ICR/f rats, a recessive-type hereditary cataractous strain. In this study, we determined the corneal penetration mechanism of DSF solutions using human cornea epithelial cell monolayers based on the immortalized human cornea epithelial cell line (HCE-T) developed by Tropainen et al. [Invest. Ophthalmol. Vis. Sci., 42, 2942-2948 (2001)]. The transepithelial electrical resistance (TER) values of HCE-T cells increases from approximately 275 to 388 Omega.cm(2) by exposure to an air-liquid interface for 2 weeks. The penetration of DSF into the basolateral chamber was prevented by the increase in TER values. The DSF in solution was converted to diethyldithiocarbamate (DDC) during the penetration experiment using HCE-T cell monolayers, and a close relationship between the penetration coefficient of DDC and aldehyde dehydrogenase (ALDH) 3A1 mRNA expression (y=41.202x+18.587, R=0.9413) was observed. In addition, an anti-ALDH3A1 antibody significantly inhibited the DSF-DDC conversion. These results suggest that DSF in DSF solutions is converted to DDC via catalysis by an ALDH3A1 in the cornea, and then transited from the apical side to the basolateral side.     

Inhibitive effects of enhanced lipid peroxidation on Ca(2+)-ATPase in lenses of hereditary cataract ICR/f rats

Our previous studies have demonstrated that the instillation of eye drops containing disulfiram, a radical scavenger and nitric oxide synthase inhibitor, delays cataract development in ICR/f rats, and we have suggested that the production of nitric oxide (NO) and lipid peroxide (LPO) in the lens may relate to the delay in cataract development brought about by disulfiram. However, the involvement of NO and LPO in lenses of ICR/f rats during cataract development has not yet been established. In the present study, we determined changes in NO and LPO levels in lenses of ICR/f rats during cataract development. Opacification of ICR/f rat lenses started at 77 days of age, and the lenses of 91-day-old ICR/f rats were almost entirely opaque. The Ca(2+)-ATPase activity in the lenses of ICR/f rats decreased with increasing age, and an elevation in Ca(2+) content was observed in ICR/f rat lenses with the decrease in Ca(2+)-ATPase activity. NO levels in the lenses of ICR/f rats increased from 63 to 85 days of age, reaching a maximum at 77 days of age. In addition, LPO levels in the lenses of ICR/f rats also increased with increasing age. LPO levels in the lenses of 63- to 91-day-old ICR/f rats were found to be significantly higher compared with those in 22-day-old ICR/f rats. These changes of Ca(2+), Ca(2+)-ATPase, NO and LPO were attenuated by instillation of DSF eye drops. These results suggest that excessive NO may cause enhanced lipid peroxidation resulting in the inhibition of Ca(2+)-ATPase. The decrease in Ca(2+)-ATPase activity may cause the elevation in lens Ca(2+), leading to lens opacification in ICR/f rats.     

细胞色素C滴眼剂的制备及药效学

目的制备细胞色素C滴眼剂,并对其基本性质与对硒致白内障大鼠的抗白内障作用进行初步研究。方法对制得的滴眼剂进行pH与澄明度、黏度与渗透压及酶活力与药效学考察。结果与结论制剂各项指标符合2000年版《中华人民共和国药典》对滴眼剂的要求,且具有良好的抗白内障活性。     

Correlation between hyper-sensitivity to hydrogen peroxide and low defense against Ca(2+) influx in cataractogenic lens of Ihara cataract rats

Our previous studies have demonstrated that lipid peroxidation in the lenses of hereditary cataract model rats (Ihara cataract rat (ICR)/f rats) caused a dysfunction in Ca(2+) regulation. In the present study, we investigated the effect of in vitro hydrogen peroxide (H(2)O(2)) stimulation on lipid peroxide (LPO) and the activities of sarco-/endoplasmic reticulum and plasma membrane Ca(2+)-ATPase (SERCA and PMCA) in the ICR/f rat lenses. An increase in LPO level and decreases in the SERCA and PMCA activities were observed with increasing H(2)O(2) concentration, and pretreatment with diethyldithiocarbamate, a potent radical scavenger, attenuated these changes in normal and ICR/f rat lenses. The glutathione levels, glutathione peroxidase and glutathione reductase activities are significantly lower in ICR/f rat lenses than in normal rat lenses. Furthermore, we presented as two kinetic parameters such as DP (defense point) and K(s) (reactive constant) analyzed from above various biological responses vs. H(2)O(2) concentration-profile curves using a one-exponential equation. The DPs for LPO, SERCA and PMCA in ICR/f rat lenses is lower than in normal rat lenses. In contrast to the results in DP, the K(s) for LPO, SERCA and PMCA in ICR/f rat lenses is higher than in normal rat lenses. In addition, the closed relationship of was observed between DP and K(s) for LPO, SERCA and PMCA. These results show that the resistance to H(2)O(2) in the ICR/f rat lenses is lower than that of normal rats. The DP and K(s) values can provide an useful information for resistances to various stimuli in cells and tissues.     

2015—2019年天津市眼科医院局部抗青光眼用药的使用情况分析

目的 分析2015—2019年天津市眼科医院局部抗青光眼药品的使用情况及发展趋势。方法 对天津市眼科医院2015—2019年局部抗青光眼药品的销售金额、用药频度（DDDs）、日均费用（DDC）、排序比（B/A）等进行回顾性统计分析。结果 前列腺素衍生物类局部抗青光眼药品的销售金额远高于其他4类药品销售金额。拉坦前列素滴眼液（国产）、拉坦前列素滴眼液（进口）、曲伏前列素滴眼液、盐酸卡替洛尔滴眼液、酒石酸溴莫尼定滴眼液、布林佐胺滴眼液稳居本院用药金额前6位。盐酸卡替洛尔滴眼液DDD稳居第1位。除了2015—2016年盐酸卡替洛尔滴眼液的DDC值略有上升外,其他滴眼液的DDC值均维持不变或呈下降的趋势。盐酸卡替洛尔滴眼液的B/A最高。结论 本院局部抗青光眼药品使用基本合理,前列腺素衍生物类药物逐渐成为抗青光眼的一线用药。     

2017—2019年天津市眼科医院抗干眼症相关药物的使用情况分析

目的 探讨2017—2019年天津市眼科医院抗干眼症相关用药的使用情况和发展趋势。方法 对2017—2019年天津市眼科医院抗干眼症相关药品的用药金额、用药频度（DDDs）、限定日费用（DDC）和药品排序比（B/A）进行统计分析。结果 人工泪液类滴眼液用药金额占到抗干眼症相关用药总金额的60%以上,聚乙烯醇滴眼液用药金额稳居第1位。重组人表皮生长因子衍生物滴眼液DDDs排序上升排位最多,复合增长率最高。2017—2019年除了羧甲基纤维素钠滴眼液的DDC值略有上升外,其他滴眼液的DDC值均呈平稳及下降趋势。0.1%玻璃酸钠滴眼液（国产）的B/A值最高,53.3%的滴眼剂B/A接近1.00。结论 天津市眼科医院抗干眼症相关药品的使用基本合理,人工泪液类滴眼液仍是抗干眼症的首选用药。     

某眼科医院2011-2012年降眼压类滴眼液利用分析

目的:了解降眼压类滴眼液的使用情况,探讨其临床用药的特点和发展趋势。方法:对某眼科医院2011-2012年11种降眼压类滴眼液的用药频度(DDDs)和日均费用(DDC)进行统计分析。结果:该院降眼压类滴眼液的DDDs逐年上升,但2011-2012的DDDs排位基本相同,排名前3位的分别是噻吗洛尔、布林佐胺和毛果芸香碱滴眼液;DDC排位前列的主要是前列腺素衍生物类滴眼液,噻吗洛尔滴眼液则DDC最低。结论:降眼压类滴眼液种类较多,价廉且疗效确切的降压类滴眼液在临床上应用中占优势,单价较高品种的DDC呈逐年下降趋势,前列腺素衍生物用量逐年增加。     

双硫伦包合物滴眼剂的制备与性质测定

目的制备双硫伦 (DSF)包合物滴眼剂。方法将DSF与HP β CD作用形成包合物 ,再制成滴眼剂。采用HPLC法测定DSF的含量。结果DSF包合物滴眼剂的 pH、渗透压、粘度及表面张力等性质 ,均符合《中华人民共和国药典》中滴眼剂的要求。滴眼剂的稳定性实验表明 :在 2 5℃ ,t0 9为 2 8a ,且在光照条件下 ,稳定性下降。结论双硫伦包合物滴眼剂达到《中华人民共和国药典》对滴眼剂的要求     

In vitro cell culture model for anti-cataract drug penetration studies

Immortalized human corneal epithelial cells (HCECs) and human lens epithelial cells (HLECs) were cultured in vitro. Cells were observed under a phase-contrast microscope and the integrity of cell monolayers was assayed by transepithelial electrical resistance (TEER) determination. The permeability of disulfiram (DSF) through a HCECs monolayer was compared with that of DSF through an excised rabbit cornea. The permeability coefficients of DSF through a HCECs monolayer and excised rabbit cornea were 29.5 +/- 4.8 x 10(-6) cm/s and 34.7 +/- 5.2 x 10(-6) cm/s, respectively. Diethyldithiocarbamate (DDC) had high permeability through HLECs monolayer with a permeability coefficient of 44.6 +/- 7.1 x 10(-6) cm/s. The cytotoxicity of DDC against HLECs was investigated using the trypan blue exclusion test. For a DDC concentration of 5 mmol/l, more than 85% cells were viable. DH3a1 mRNA was expressed in cultured HLECs. The expression of aldehyde dehydrogenase 3a1 (ALDH3a1), which may be be responsible for DSF-DDC conversion, was detected using RT-PCR and agarose gels electrophoresis. These results demonstrate that the permeability of DSF can be detected and intra-ocular drug action may be predicted using the cultured HCEC and HLEC monolayers as model.     

Effects of lipid composition on the transcorneal penetration of liposomes containing disulfiram, a potential anti-cataract agent, in the rabbit

We previously demonstrated that topical treatment with disulfiram (DSF) prevented the development of cataracts in sodium selenite-injected rat pups. In biological systems, DSF is rapidly reduced to diethyldithiocarbamate (DDC), a potent antioxidant. In this study, we investigated the effect of altering the lipid composition of liposomes containing DSF on the transcorneal transit of DDC. Liposomes containing DSF were prepared with various molar ratios of dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC) and cetylpyridinum chloride (CPC) by reverse-phase evaporation. Liposomes with a DMPC to DPPC molar ratio of 5:5, examined by differential scanning calorimetry, had the highest enthalpy of transition and the presence of one molar ratio of CPC further enhanced the enthalpy value. The addition of bovine serum albumin or a homogenate of rabbit cornea to the incubation buffer resulted in the release of DDC, but not DSF from the liposomes. The amount of DDC present in the aqueous humor of rabbit eyes following topical administration increased with increase in DMPC to DPPC ratios and was also enhanced by the addition of CPC to the liposomes. The results of this study suggest that liposome formulations are effective for transcorneal drug delivery of anticataract agents such as DSF. DSF in liposomes consisting of DMPC, DPPC, and CPC with a molar ratio of 8:2:1 may be a potential drug formulation for the prevention and/or treatment of cataracts.     

Diethylthiocarbamic acid methyl ester. A potent inhibitor of aldehyde dehydrogenase found in rats treated with disulfiram or diethyldithiocarbamic acid methyl ester

Rats were treated with disulfiram (Antabuse, DSF) or its metabolite diethyldithiocarbamic acid methyl ester (Me-DDC) and challenged with ethanol. The blood pressure response to ethanol was followed and blood was analyzed for DSF, Me-DDC and diethyldithiocarbamic acid (DDC). The rat liver aldehyde dehydrogenase (ALDH) isozyme activities were measured 2 hr after the ethanol challenge. Both treatments produced a significant fall in the blood pressure when challenged with ethanol, probably caused by a marked decrease in hepatocyte low Km and high Km activities. The mean plasma concentration ranges of Me-DDC and DDC were found to be 49-1241 nmol/l and 182-841 nmol/l, respectively, whereas DSF was undetectable. In addition, it was found that inactivation of hepatocyte low Km ALDH activity was dependent on preoxidation of Me-DDC by the microsomal cytochrome P-450 mixed function oxidases. Me-DDC was found to be oxidized under aerobic conditions in the presence of NADP to form diethylthiocarbamic acid methyl ester (Me-DTC). The structure was confirmed from its MS/EI fragmentation spectrum. Me-DTC was found to be a potent inhibitor of low Km ALDH when added to rat liver homogenate. The compound was also identified as a metabolite in rat blood collected from the DSF and Me-DDC treated rats, and in blood from human alcoholics on DSF treatment. Me-DTC appears to be more selective for the low Km isozymes whereas the opposite seems to be the case for the hydrolytic product, DTC.     

壳聚糖对阿昔洛韦滴眼液离体角膜渗透促进作用的考察

目的：考察壳聚糖（CS）对阿昔洛韦（ACV）滴眼液（ACV-CS）离体角膜渗透促进作用,为研制新型ACV滴眼液奠定基础.方法：以CS为增黏剂及吸收促进剂,制备ACV-CS滴眼液;以不加CS的ACV滴眼液为对照,采用Franz扩散池法考察CS对ACV滴眼液离体角膜渗透促进作用.结果：与不加ACV滴眼液相比,ACV-CS滴眼液的黏性显著增加;给药20 min后,ACV-CS滴眼液对离体角膜的累积渗透药量明显大于不加CS的ACV滴眼液（P＆lt;0.05）.结论：CS对ACV滴眼液离体角膜渗透具有明显的促进作用,值得进一步研究.     

复方托吡卡胺滴眼液致突发性耳聋

1名60岁女性糖尿病患者，行眼底检查前用复方托吡卡胺滴眼液散瞳，随后出现听力明显下降伴耳呜，检查提示双耳感音神经性耳聋。经高压氧、葛根素治疗后，听力明显恢复。因眼科检查需要，再次用复方托吡卡胺滴眼液，用药后患者又出现听力明显下降，经对症治疗后好转。当患者第4次使用该药后出现耳聋，治疗1月余，听力仍未恢复。     

拉坦前列素滴眼液诱发支气管哮喘

1例65岁女性患者，因右眼青光眼给予拉坦前列素滴眼液0．1ml，1次／d点眼，首次用药2h后患者出现呼吸困难、发绀、大汗淋漓，HR120次／min，P160／100mmHg，双肺闻及哮鸣音。立即给予患者沙美特罗替卡松粉吸入剂，10min后症状缓解。以后改用1％布林佐胺滴眼液，支气管哮喘未再出现。     

Serum albumin and the metabolism of disulfiram

The effectiveness of tetraethylthiuram disulfide (DSF) as a drug used in the treatment of alcohol abuse has been limited by the fact that it is degraded rapidly in the tissues and in the serum. Hence, a useful dose-response curve for this drug cannot be determined easily. The degradation in the tissues has been well characterized; however, its fate in the serum is less well understood. Here we kinetically describe the first steps in the degradation of DSF in the serum which results from a covalent interaction of this drug with the free sulfhydryl of serum albumin. DSF and its cleavage product diethyldithiocarbamate (DDC) both absorb significantly in the ultraviolet region. The reduction of DSF with mercaptoethanol to two molecules of DDC resulted in a large change in absorption in this region. The reaction of serum albumin with DSF produced a similar but much slower change in the ultraviolet absorption. As a result of the existence of this slow spectral change, we have been able to directly and continuously monitor the interaction of serum albumin and DSF and have determined that it is an overall first-order process. A model is proposed wherein DSF and serum albumin rapidly form a noncovalent adduct and, subsequently, in a slow unimolecular process, DSF is reduced to one mole of free DDC and one mole of the serum albumin-DDC mixed disulfide. At pH 9 the half-time for this process was 30 to 40 sec, and at pH 7.4 the half-time for this process was 1 to 1.5 min. These results suggest that degradation of DSF by serum albumin is physiologically and clinically important since the drug is maximally active only many hours after administration.     

Topical Lomefloxacin versus Topical Chloramphenicol in the Treatment of Acute Bacterial Conjunctivitis

We performed a prospective, randomised, investigator-masked and parallel-group study to compare topical lomefloxacin 0.3% instilled twice daily with topical chloramphenicol instilled five times daily in the treatment of acute bacterial conjunctivitis. 191 patients (lomefloxacin 96, chloramphenicol 95) were enrolled in this study with clinically diagnosed acute bacterial conjunctivitis. The two treatment groups were similar at baseline. The treatments were equally effective and significantly (p < 0.001) reduced the Cumulative Sum Score of the clinical signs and symptoms of bacterial conjunctivitis. At the end of the trial, there was no difference between the two treatments in the Cumulative Sum Score of signs and symptoms (p = 0.63), and the investigator (p = 0.28) and patients' (p = 0.50) assessments of the success of therapy. The two drugs were equally well tolerated locally, with no serious systemic or local adverse drug reactions reported in any study patient. Bacteriological confirmation of acute conjunctivitis was possible in 96 patients (lomefloxacin 47, chloramphenicol 49) out of the 191 enrolled. Both treatments significantly (p < 0.001) reduced the conjunctival bacterial colony count score with no difference (p = 0.12) between the two treatment groups. In conclusion, lomefloxacin 0.3% eye drops instilled twice daily were as effective and well tolerated as chloramphenicol 0.5% eye drops instilled 5 times daily in the treatment of acute bacterial conjunctivitis.     

The local vasoconstriction of infant’s skin following instillation of mydriatic eye drops

Background  

Systemic absorption of eye drops is known to occur via the nasal mucosa, cornea, and conjunctiva. Diffusion of eye drops through the skin is previously unrecognized. Here, two cases are presented in which we observed skin pallor around the eyes after instillation of phenylephrine 2.5% drops. Case 1 A 32-week gestational age premature infant had mydriatic eye drops instilled as part of retinopathy of prematurity screening. Case 2 A term newborn dysmorphic infant underwent fundus examination to rule out ocular pathology. In both cases, discoloration of periorbital skin was observed 45 min following administration of drops.