YAKUGAKU ZASSHI
 Abstracts 
Vol.124, No. 11 (2004)

--Reviews--

Wilson's Disease and Its Pharmacological Treatment

Hisao HAYASHI,* Rie SUZUKI, and Shinya WAKUSAWA

Department of Medicine, Faculty of Pharmaceutical Sciences of Hokuriku University, Ho-3 Kanagawa-machi, Kanazawa 920-1181, Japan

Wilson's disease is an inherited copper toxicosis caused by defective putative copper transporting ATPase in the liver. Because of impaired biliary secretion, copper remains in the liver, resulting in chronic hepatic lesions including fatty metamorphosis, chronic hepatitis and cirrhosis. In the latter stage, extrapyramidal syndromes may develop with and without symptomatic hepatic lesions. Acute liver damage associated with hemolysis and deep jaundice may be the first manifestation. The majority of patients show hypoceruloplasminemia, which has been used as a screening test for the disease. A large number of mutations in the ATP7B gene have been reported. Thus, genetic diagnosis might be limitedly used to presymptomatic diagnosis of siblings when mutations are identified in an index patient. Introduction of penicillamine caused a revolution in the treatment of patients. Another chelater, trientine, is now available for those intolerant of penicillamine. Tetrathiomolibdate and zinc acetate are additional alternatives currently being tested. Hypoceruloplasminemia and further reduction after chelation therapy may be associated with iron overload. This complication is closely related with impaired transport of ferrous ion due to ferroxidase deficiency. Noncompliance and teratogenicity are other major concerns because any treatment with the agents listed above is a life long regimen. Despite various side effects of penicillamine, its teratogenicity is negligible. These data indicate that penicillamine is the first choice of drug for this disease.

Key words--ATP7B; copper chelation; iron accumulation

--Reviews--

Synthesis of Phosphonic Acids and Phosphinic Acids Derivatives toward to Biologically Active Compounds

Shiroshi SHIBUYA

School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji 192-0392, Japan

This paper covers recent publications from our laboratory on the synthesis of a variety of phosphonate and phosphinate derivatives. New methods for the enantioselective synthesis of α-hydroxyphosphonates were established by Lewis acid-mediated cleavage of homochiral 1,3-dioxaneacetals with P(OEt)₃ and chiral metal ligand-mediated hydrophosphonylation of aldehydes. Two diastereomers of HPmp derivatives were prepared by an application of these methods. The HPmp derivatives were convered to FPmp derivatives but with low diastereoselectivity. Hydrophosphonylation of α-aminoaldehydes afforded threo- and erythro-β-amino-α-hydroxyphosphonates under chelation and nonchelation controlled conditions, respectively. The asymmetric dihydroxylation of α, β-, and β, γ-unsaturated phosphonates with AD-mix-α and AD-mix-β reagents gave α, β-, and β, γ-dihydroxyphosphonates with high enantioselectivity. The method was applied to the kinetic resolution of racemic α-oxygetated β, γ-unsaturated phosphonates. Treatment of allyloxymethylphosphonates with the base afforded α-hydroxyphosphonates via the [2,3]-Wittig reaction. Threo- and erythro-β-amino-α-hydroxyphosphinates were obtained with high diastereoselectivity by phosphinylation of α-aminoaldehydes in the presence of (R)- and (S)-ALB, respectively. The phosphinylation of α-oxygenated aldehydes afforded the corresponding α, β-dioxygenated phosphinates, but with low diastereoselectivity. Sphingomyelin analogues containing CF₂PO(OH)₂ were synthesized starting from (S)- and (R)-Garner aldehyde for the purpose of obtaining potent sphyngomyelinase inhibitors. A useful method for the synthesis of α, α-difluorobenzylphosphonates was established based on the cross-coupling reaction of an iodobenzene derivative with ZnCuBr₂CF₂PO(OEt)₂. The synthetic utility of ZnCuBr₂CF₂PO(OEt)₂ was examined to obtain α, α-difluoromethylenenphosphonates. The method was applied to the synthesis of PNP-inhibitory active compounds by combination of the purine base and alcohols containing difluoromethylenephosphonate. The methodology for the β-selective N-glycosylation of 2,3-dideoxy glucoside was established by introducing phosphonothioates at the 3-position of glycosyl doners instead of phosphonate. Synthesis of new acylic nucleotide analogues designed based on the structural modification of ARS2267 is also described. Finally, kiral synthesis of some phosphonates was achieved using lipase through kinetic resolution.

Key words--phosphonate; phosphinate; bioisoster; α, α-difluoromethylenephosphonate; nucleotide analogue; N-glycosilation

--Reviews--

X-Ray Crystallographic Analysis of Picrates

Hiroaki TAKAYANAGI,*^,a Motoaki GOTO,^b Kazuyoshi TAKEDA,^c and Yumiko OSA^a

School of Pharmaceutical Sciences, Kitasato University,^a 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan, Saitama-Daiichi Pharmaceutical Co., Ltd.,^b 8-1 Minamisakae, Kasukabe 344-0057, Japan, and Ebara Research Co., Ltd.,^c 4-2-1 Honfujisawa, Fujisawa 251-8502, Japan

Picric acid forms stable picrates with various organic molecules through π- bonding or ionic bonding, and such picrates have been very useful for identification and qualitative analysis. As it seemed desirable to determine the crystal structures and the bonding mode of picrates of basic organic compounds, we have investigated the crystal structures of aromatic hydrocarbons, aromatic amino compounds, heterocyclic compounds and so on. A series of our studies on the crystal structure of basic organic compounds have shown that the complexes of picric acid and aromatic hydrocarbons are formed through π-bonding, and those of aromatic heterocyclic compounds are formed through ionic and hydrogen bonding; in addition, some of them also have π-bonding.

Key words--x-ray analysis; picrate; crystal structure; aromatic hydrocarbon; aromatic amine; heterocyclic compound

--Reviews--

Development of Novel DDS Technologies for Pharmacoproteomic-based Drug Discovery and Development

Yasuo TSUTSUMI¹⁾

Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita 565-0871, Japan

With the success of the Human Genome Project, the focus of life science research has shifted to the functional and structural analyses of proteins, such as proteomics and structural genomics. These novel approaches to the analysis of proteins, including newly identified ones, are expected to help in the identification and development of protein therapies for various diseases. Thus pharmacoproteomic-based drug discovery currently has a very high profile. Nevertheless, the use of bioactive proteins in the clinical setting is not straightforward because in vivo these proteins have low stability and pleiotropic action. To promote pharmacoproteomic-based drug discovery and development, we have attempted to establish a system for creating functional mutant proteins (muteins) with the desired properties and to develop a site-specific bioconjugation system for further improving their therapeutic potency. These innovative protein-drug systems are discussed in this review.

Key words--phage display system; proteomics; bioconjugation; protein therapy; targeting

--Reviews--

Bridged Nucleic Acids (BNAs) as a Basic Material for Genome Technology

Satoshi OBIKA

Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita 565-0871, Japan

The completion of the human genome sequencing project will greatly accelerate the development of novel and practical technologies for genome-analysis, diagnostics or therapeutics. Oligonucleotides are playing an important role in these genome technologies, because of their sequence-specific hybridization ability toward the complementary strand. Besides the sequence-specific duplex formation, oligonucleotides are able to form stable triplex structures, which is fundamental to the antigene strategy to regulate gene expression in a living cell. However, two major drawbacks are known in the triplex formation by a natural oligonucleotide: low stability of the triplex and limitations of the target DNA sequence. One promising strategy to overcome these problems is chemical modification of the oligonucleotides. We have developed various bridged nucleic acids (BNAs), and found that the oligonucleotides containing 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) modification form a stable parallel motif triplex with the double-stranded DNA target under physiological conditions. Some nucleobase analogues to extend the target DNA sequence were designed, synthesized and incorporated into the 2',4'-BNA structure. The obtained 2',4'-BNA derivatives containing modified nucleobases effectively recognized a pyrimidine-purine interruption. Some other examples of nucleic acid analogues to overcome the two major drawbacks in the triplex-forming oligonucleotides are also summarized.

Key words--nucleic acid analogue; triplex; antigene; sugar conformation; bridged nucleic acid; locked nucleic acid

--Reviews--

The Physiological Function of the Blood-Brain Barrier Transporters as the CNS Supporting and Protecting System

Sumio OHTSUKI

Graduate School of Pharmaceutical Sciences, and New Industry Creation Hatchery Center, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan, SORST and CREST, Japan Science and Technology Agency, Japan

The blood-brain barrier (BBB) segregates the circulating blood from interstitial fluid in the brain and restricts drug permeability into the brain. Our latest studies have revealed that the BBB transporters play important physiological roles in maintaining the brain environment. For an energy-storing system, the creatine transporter localized at the brain capillary endothelial cells (BCECs) mediates the supply of creatine from the blood to the brain. The BBB is involved in the brain-to-blood efflux transport of γ-aminobutyric acid, and GAT2/BGT-1 mediates this transport process. BCECs also express serotonin and norepinephrine transporters. Organic anion transporter 3 (OAT3) and ASCT2 are localized at the abluminal membrane of the BCECs. OAT3 is involved in the brain-to-blood efflux of a dopamine metabolite, a uremic toxin, and thiopurine nucleobase analogues. ASCT2 plays a role in L-isomer-selective aspartic acid efflux transport at the BBB. Dehydroepiandrosterone sulfate and small neutral amino acids undergo brain-to-blood efflux transport mediated by organic anion transporting polypeptide 2 and ATA2, respectively. The BBB transporters are regulated by various factors: ATA2 by osmolarity, taurine transporter by tumor necrosis factor-α, and L-cystine/L-glutamic acid exchange transporter by oxidative stress. Clarifying the physiological roles of BBB transport systems should give important information allowing the development of better central nervous system (CNS) drugs and improving our understanding of the relationship between CNS disorders and BBB function.

Key words--blood-brain barrier; transporter; brain capillary endothelial cells; creatine; neurotransmitter; neuro-modulator

--Reviews--

Design and Synthesis of Subtype- and Species-Selective Peroxisome Proliferator-Activated Receptor (PPAR) Alpha Ligands

Hiroyuki MIYACHI

Laboratory of Bioorganic Chemistry, Department of Structural Biology, Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan

The molecular pharmacological discovery of the nuclear receptor peroxisome proliferator-activated receptor alpha (PPARα) as the master regulator of lipid and lipoprotein homeostasis, and the rapid development of a parallel screening approach to evaluate activity towards other PPAR subtypes (PPARδ, and PPARγ) have provided an opportunity to develop novel PPARα-selective, PPARα/γ dual, and PPAR pan agonists. This review focuses on the molecular pharmacology of PPARα, and summarizes our current design, synthesis, and evaluation of subtype-selective PPARα agonists. The species selectivity of several classes of PPARα selective agonists in response to in vitro PPARα transactivation activity is also reported. These studies should help us to understand the structure-activity relationships and the mode of interaction between ligands and PPARα, and also help to create novel therapeutic choices for the treatment of metabolic disorders.

Key words--peroxisome proliferator-activated receptor (PPAR); PPARα; species selectivity

--Regular Articles--

Comparison of the Effects of Prophylactic Antibiotic Therapy and Cost-effectiveness between Cefazolin (CEZ) and Sulbactam/Ampicillin (SBT/ABPC) in Gastric Cancer Surgery Employing Clinical Pathway

Yuya ISE,^*,a Ken HAGIWARA,^a,d Setsuo SAITOH,^d Kazuyoshi HONJO,^a Shizuka SOH,^a Ayumi KATO,^a Shirou KATAYAMA,^a Kenji NISHIZAWA,^a Masaaki HIRANO,^a Toshiro YOSHIYUKI,^c Teruo KIYAMA,^c Kyoko MITSUHASHI,^b Miwako KAMEI,^e and Makoto SHIRAGAMI^e

Department of Pharmceutical Service,^a Department of Nursing,^b Department of Surgery (I),^c Nippon Medical School Hospital, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8603, Japan, Faculty of Pharmaceutical Sciences, Josai University,^d 1-1 Keyakidai, Sakado 350-0290, Japan, and Social and Administrative Pharmacy Sciences, College of Pharmacy, Nihon University,^e 7-7-1 Narashinodai, Funabashi 274-8555, Japan

The present study was designed to investigate the effects of prophylactic antibiotic therapy and the cost-effectiveness of Cefazolin (CEZ) and Sulbactam/Ampicillin (SBT/ABPC) in gastric cancer surgery employing clinical pathway. 157 patients (62 in the CEZ group and 95 in the SBT/ABPC group), who underwent surgery for gastric cancer at the First Department of Surgery of our hospital, were investigated. There was no significant difference between the groups with regard to sex, age, incidence of complication, stage of cancer, surgical method, operative time and blood loss, length of hospitalization, the appearance of systemic inflammatory response syndrome (SIRS), changes body tempreture, white blood cell count (WBC), C-reactive protein (CRP), or clinical outcome of postoperative care by a nurse during post-operation for 7 days. The prophylactic effect of infection was also no different between the CEZ (69.4%) and SBT/ABPC (69.5%) groups. In contrast, decision analysis strongly indicated that the anticipate cost of antibiotics was higher in the latter group (¥20402) than in the CEZ group (¥15556), suggesting that the prophylactic effect of CEZ may be more cost-effective. Thus, evaluations of pharmacotherapy from the aspect of cost may be one of the important responsibility of hospital pharmacists in the future.

--Regular Articles--

β-Adrenergic Blocking Agents and Intermittent Claudication: Systematic Review

Ritsuko MIYAJIMA,*^,a,b Kazumi SANO,^b and Hisahiro YOSHIDA^b

Course of Clinical Pharmacy, Graduate School,^a and Department of Drug Metabolism and Disposition,^b Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose City Tokyo 204-8588, Japan

To clarify contradictions in past reports and the package inserts for β-adrenergic blocking agents (β-blockers) for patients with intermittent claudication (IC), we investigated the effects of β-blockers in patients with IC using the systematic review technique. Data sources were randomized, controlled trials that investigated the effects of β-blockers compared with the placebo or untreated group (controls) in patients with IC. Primary endpoints were walking distance and walking time, and secondary endpoints were ankle-brachial index (ABI) and calf blood flow. Nine trials were included in the analysis. Meta-analysis showed that there was a significant worsening in maximal walking distance and initial claudication distance in patients receiving β-blockers, with standardized mean differences of −0.31 and −0.39 (95% confidence interval −0.58 to −0.04 and −0.73 to −0.06, P=0.03 and 0.02, respectively) compared with controls. There were no significant differences in maximal walking time (0.07, −0.24 to 0.37), time to onset of claudication (0.12, −0.23 to 0.47), ABI at rest (0.24, −0.30 to 0.78), calf blood flow at rest (0.00, −0.26 to 0.25), and calf blood flow after exercise (−0.23, −0.69 to 0.22). However, only one trial evaluated ABI, and the number of cases is increasing, suggesting that β-blockers do not worsen ABI. There was no evidence that β-blockers prescribed for patients with IC have unsuitable “precautions” in the package inserts. However, reluctance to administer β-blockers to patients because they have IC is not appropriate.

Key words--intermittent claudication; peripheral vascular diseases; adrenergic beta-antagonists; meta-analysis

--Regular Articles--

Pharmaceutical and Pharmacological Evaluation of Burow's Solution (Aluminum Acetate Solution), a Hospital Preparation, and Development of Its Rapid Preparation Method

Yuko ISHIBASHI,^a Teruo MURAKAMI,^a Ryoko YUMOTO,^a Masahiko SAKAI,^b Hiromichi SHINTANI,^b Hideyuki ITAHA,^c Masayuki KANBE,^c Shin MASUDA,^d Koji YAJIN,^d Kenji KIHIRA,^b and Mikihisa TAKANO*^,a

Department of Pharmaceutics and Therapeutics,^a Department of Otorhinolaryngology,^d Graduate School of Biomedical Sciences, Hiroshima University, Department of Pharmaceutical Services,^b Clinical Pathology,^c Hiroshima University Hospital, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan

Burow's solution, or aluminum acetate solution, is effective in inhibiting various microorganisms including methicillin-resistant Staphylococcus aureus (MRSA) that are commonly observed in chronic suppurative otitis media. It takes several days to prepare Burow's solution using aluminum sulfate, and the pharmaceutical properties of the solution are not fully understood. In this study, the effect of storage (5 months) of Burow's solution prepared according to the Teine-Keijin Hospital manual on its pharmaceutical properties and antibacterial activities was examined. We also attempted to develop a rapid preparation method of aluminum acetate (or 1.7% aluminum) solution using two commercially available compounds of aluminum acetate basic (Al₂O(CH₃CO₂)₄, Al(OH)(CH₃CO₂)₂). The properties of Burow's solution, pH, osmolarity and antibacterial activity, were the same among different preparations and its storage for 5 months at 4°C had no effect on these properties. The antibacterial potency of Burow's solution was dependent on aluminum concentration and its antibacterial potency against S. aureus and several MRSA strains was of the same magnitude. In a rapid preparation, aluminum acetate basic was mixed with appropriate amounts of tartaric acid and acetic acid, and the suspension was boiled for 2-2.5 hr until dissolved. The rapidly prepared aluminum acetate solution showed the same pharmaceutical properties and antibacterial activities as those of Burow's solution. The newly developed preparation method for aluminum acetate solution is expected to be convenient and feasible for hospital treatment of chronic suppurative otitis media.

Key words--Burow's solution; stability; antibacterial activity; pharmaceutical property; rapid preparation; aluminum acetate solution

--Regular Articles--

Effects of Long-Term Administration of Caffeine on Fat Storage in Ovariectomized Rats

Li-Kun HAN,* Fumi KAI, and Hiromichi OKUDA

Department of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Kumamoto 862-8502, Japan

It is well known that obesity occurs in women with climacteric disorders. The aim of this study was to examine whether caffeine prevents obesity and bone loss in ovariectomized rats (ovx). Eight-week-old female Wistar rat were assigned to 4 groups: a sham-operated group fed the control diet (CE-2); an ovx-c group fed the control diet; an ovx-caf 0.15% group fed the control diet containing 1.5 g/kg of caffeine; and an ovx-caf 0.3% group fed the control diet containing 3 g/kg of caffeine. Body weights at 2-9 weeks and the final parametrail adipose tissue weights were significantly lower in the ovx-caf 0.3% group than in the ovx-c group. Food intakes were significantly lower in the ovx-caf 0.3% group than in the ovx-c group. After 9 weeks, the rats were killed and adipose tissues were sampled immediately. Basal lipolysis was increased in the ovx-caf 0.3% group fed the control diet containing 3 g/kg of caffeine than in the ovx-c group fed the control diet. The relative content of calcium (g/100 g body weight) in the ovx-caf 0.3% group was significantly increased compared with that in the ovx-c group. These results show a new possible role for caffeine in the prevention of lifestyle-related diseases.

Key words--caffeine; basal lipolysis; ovariectomized-rats

--Notes--

Assessment of Antioxidant Activity of Natural Compound by Water- and Lipid-soluble Antioxidant Factor

Eiji USAMI,*,^a Genjiro KUSANO,^b Takanori KATAYOSE,^c Hiroshi WACHI,^c and Yoshiyuki SEYAMA^c

Chigasaki Municipal Hospital,^a 5-15-1 Honson, Chigasaki 253-0042, Japan, Osaka University of Pharmaceutical Sciences,^b 4-20-1 Nasahara, Takatsuki 569-1094, Japan, and Faculty of Pharmaceutical Sciences, Hoshi University,^c 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501, Japan

We evaluated the antioxidant activity of natural compounds in water-soluble and lipid-soluble phases and found that ferulic acid, quercetin and caffeic acid showed stronger activity in the water-soluble phase. Various fractions isolated from Bidens pilosa showed this activity mainly in the water-soluble phase. Antioxidant activity in the lipid-soluble phase of propolis depended on the lipophilic extraction.

Key words--antioxidant activity; Bidens pilosa; propolis

--Notes--

Studies on Antihypertensive Effect of Luobuma (Apocynum venetum L.) Leaf Extract (3)

Chie TAGAWA,*^,a Tamami KAGAWA,^b Yoshihisa NAKAZAWA,^a Shigenori ONIZUKA,^a Sansei NISHIBE,^c and Hiromu KAWASAKI^d

Hitachizosen Corpolation,^a 1-2264 Habu-cho, Innoshima City, Hiroshima 722-2393, Japan, Dydo DRINCO, INC.,^b 6F-2-8-3 Shiba, Minato-ku, Tokyo 105-0014, Japan, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido,^c Ishikari-Tobetsu, Hokkaido 061-0293, Japan, and Department of Clinical Pharmaceutical Science, Graduate School of Natural Science and Technology, Okayama University,^d 1-1-1 Tsushima-naka, Okayama 700-8530, Japan

To clarify the mechanisms underlying the antihypertensive effect of Luobuma (Apocynum venetum L. (Apocynaceae)) leaf extract (LLE), we investigated the vasodilator effect of LLE in the rat mesenteric vascular bed, which plays an important role in changes in peripheral resistance and thus the regulation of blood pressure. In the perfused mesenteric vascular bed with active tone and intact endothelium, perfusion of LLE (0.1 ng to 100 mg/ml for 15 min) caused dose-dependent vasodilation, which was abolished by chemical removal of the endothelial layer with perfusion of sodium deoxycholate, but not by N^G-nitro-L-arginine-methyl ester (L-NAME), a competitive inhibitor of nitric oxide (NO), which instead increased the effect. The LLE-induced vasodilation was partially inhibited by high K⁺-containing Krebs solution and tetraethylammonium (a K⁺ channel blocker) and completely by the combination of L-NAME and high K⁺-Krebs solution. However, atropine (a muscarinic acetylcholine receptor antagonist) did not affect the vasodilation. These results suggest that the vasodilation induced by LLE is endothelium-dependent and mediated by endothelium-derived hyperpolarizing factor, which involves the activation of K⁺-channels. The higher concentrations of LLE may enhance NO production/release to cause vasodilation.

Key words--Apocynum venetum L.; Luobuma leaf extract; antihypertensive effect; endothelium-dependent relaxation; rat mesenteric vascular bed