Activities of various 6-chloro-6-deoxysugars and (S) alpha-chlorohydrin in producing spermatocoeles in rats and paralysis in mice and in inhibiting glucose metabolism in bull spermatozoa in vitro

6-Chloro-6-deoxyglucose, 6-chloro-6-deoxymannose, 6-chloro-6-deoxy-fructose, 6-chloro-6-deoxyglucitol, 6-chloro-6-deoxygalactose and (S) alpha-chlorohydrin all produced spermatocoeles in the ductuli efferentes and epididymis of the rat and were neurotoxic in the mouse, but only alpha-chlorohydrin caused substantial inhibition of glucose metabolism in bull spermatozoa in vitro. The relative potencies of the compounds in producing spermatocoeles reflected their activities as reversible antifertility agents in the rat but compared to the others 6-chloro-6-deoxymannose was considerably less neurotoxic to mice than might have been anticipated from its contraceptive dose. Thus different metabolites may be responsible for causing the antifertility and the neurotoxic effects.     

Control of the Indian mole rat with alpha-chlorohydrin: laboratory studies on bait acceptance and antifertility effects

Toxic and antifertility effects of feeding poison baits of a toxicant-sterilant, alpha-chlorohydrin, were studied against the Indian mole rat Bandicota bengalensis . It was found that 0.5% alpha-chlorohydrin bait was the most palatable formulation which delivered the amount of active ingredient equal to or more than MLD (82 mg/kg) to B. bengalensis in a single day's feeding. The rats suffered maximum mortality with this bait concentration both in no-choice and bi-choice feeding trials. Male survivors of 0.5% and 1.0% alpha-chlorohydrin baits showed functional abnormalities of their testes as revealed by loss in testicular weight, decrease in the diameter of seminiferous tubules and thickness of seminiferous epithelium and abnormally low levels of spermatogenic cells. Effect of the poison on epididymis became apparent by the presence of epididymal lesions in caput epididymides and low levels of sperm concentration, live sperms and sperm motility in the cauda epididymal fluid. Our findings on the acceptance and toxiccum-antifertility effects of feeding alpha-chlorohydrin baits suggest field evaluation of this poison for the management of B. bengalensis would be appropriate.     

The effects of alpha-chlorohydrin on the composition of rat and rabbit epididymal plasma: a possible explanation of species difference.

The relationship between the antifertility effect of alpha-chlorohydrin and changes in composition of luminal plasma from the cauda epididymidis of rats and rabbits has been investigated. At each dose regimen studied, the fertilizing capacity of rats treated with alpha-chlorohydrin was reduced to zero. The levels of sodium, potassium, glycerylphosphorylcholine (GPC), acid phosphatase and alkaline phosphatase in epididymal plasma were not markedly affected by drug treatment. The most noticeable change was a considerable increase in the concentration of lactic dehydrogenase (LDH) at all dose levels and of glutamic-oxaloacetic transaminase (GOT) after 7 days of treatment with 8 and 16 mg/kg. The effect of cold shock on the composition of epididymal plasma showed that LDH and GOT are, at least in part, derived from spermatozoa. In contrast, alpha-chlorohydrin did not have an antifertility action in the rabbit, and the only notable change in the compositon of epididymal plasma was an increase in the level of GPC. These results provide evidence that, in the rat, alpha-chlorohydrin or a metabolite primarily exerts its antifertility effect by a direct action on the spermatozoa, whilst in the rabbit a barrier may exist to the entrance of the drug into the lumen of the epididymal duct.     

Inhibition of fructolytic enzymes in boar spermatozoa by (S)-alpha-chlorohydrin and 1-chloro-3-hydroxypropanone

When boar spermatozoa were incubated with the (S)-isomer of the male antifertility agent alpha-chlorohydrin the activity of glyceraldehyde-3-phosphate dehydrogenase was inhibited. The (R)-isomer had no significant effect on the activity of this enzyme whereas (R,S)-3-chlorolactaldehyde caused an inhibition of its activity and also in that of lactate dehydrogenase. The in vitro production of (S)-3-chlorolactaldehyde, the active metabolite of (S)-alpha-chlorohydrin, was attempted by incubating boar spermatozoa with 1-chloro-3-hydroxypropanone. Preliminary results lead us to propose that this compound is converted into (S)-3-chlorolactaldehyde as well as to another metabolite which is an inhibitor of other enzymes within the fructolytic pathway.     

Laboratory evaluation of a toxicant-sterilant, alpha-chlorohydrin, for the control of Indian mole rat Bandicota bengalensis

A toxicant-sterilant, alpha-chlorohydrin, was evaluated against the Indian mole rat Bandicota bengalensis and the ship or house rat Rattus rattus. It caused 100% mortality of B. bengalensis at 100 mg/kg but no mortality was observed in R. rattus even at 300 mg/kg. The acute oral LD₅₀ for B. bengalensis was found to be 82 mg/kg. The survivors of B. bengalensis, which received 60 - 90 mg/kg of alpha-chlorohydrin by oral intubation, showed dose-dependent decreases in testicular weight, and cauda epididymal sperm concentration, live sperm count and sperm motility. The values of these parameters indicated that mole rats receiving the above doses would be sterile. In contrast, these changes were observed in R. rattus only at doses of 200 and 300 mg/kg. The toxic and antifertility effects of alpha-chlorohydrin observed on B. bengalensis suggest that it should be evaluated for the management of this species under held conditions.     

Genome-wide profiling of gene expression in the epididymis of alpha-chlorohydrin-induced infertile rats using an oligonucleotide microarray

Background  

As one of the chlorinated antifertility compounds, alpha-chlorohydrin (ACH) can inhibit glyceraldehyde-3-phosphate dehydrogenase (G3PDH) activity in epididymal sperm and affect sperm energy metabolism, maturation and fertilization, eventually leading to male infertility. Further studies demonstrated that the inhibitory effect of ACH on G3PDH is not only confined to epididymal sperm but also to the epididymis. Moreover, little investigation on gene expression changes in the epididymis after ACH treatment has been conducted. Therefore, gene expression studies may indicate new epididymal targets related to sperm maturation and fertility through the analysis of ACH-treated infertile animals.     

Factors involved in the maintenance of luteal function in the pseudopregnant rat

Some data on a new antispermatogenic and antifertility compound, DL-6-(N-alpha-pipecolinomethyl)-5-hydroxy-indane maleate (PMHI) are presented. It was found that PMHI depressed testicular weight and interfered with spermatogenesis and fertility in the male rat. At equal doses, by weight, PMHI caused a greater reduction in testicular weight than WIN 18446, nitrofurazone, methallibure, and diethylstilbestrol, but lesser reductions in seminal vesicle and ventral prostate weights than were obtained with the 2 antigonadotropic compounds, methallibure and diethylstilbestrol. The data obtained in the experiments indicate that administration of PMHI causes sterility in male rats as a consequence of antispermatogenic activity. At low dosage levels (3 mg/kg), the action was reversible, but animals given 6.25 mg/kg/day or more remained sterile for 19 weeks following the treatment period. Doses of the compound which were effective in the male rat did not interfere with normal estrous cycles or fertility when administered to female rats.     

Embryopathies due to spermatozoal impairment in Xenopus laevis.

An in vitro test system, involving gametes of Xenopus laevis has been used to study the effect of antifertility agents upon spermatozoa as manifest in developing embryos. Exposure to either the isomeric forms of dimethylmyleran, methyl methanesulphonate or gamma-radiation led to development of a range of embryopathies, whereas treatment with steroidal drugs or the rodent epididymal chemosterilants alpha-chlorohydrin and trimethylphosphate was compatible with production of apparently normal offspring.     

Effect of sodium nitrite on the alpha-chlorohydrin-induced lesion of the testis--epididymis complexin the rat.

Administration of vasodilator, sodium nitrite (20 mg/kg body weight), 30 min before alpha-chlorohydrin treatment (90 mg/kg body weight) prevented the chlorohydrin-induced lesion in rat testis--epididymis complex. However, administration of vasodilator 90 min after alpha-chlorohydrin treatment did not prevent the chlorohydrin-induced lesion in the testis--epididymis complex. These observations suggest that the testicular vasculature is involved in drug action.     

动态吸附与气相色谱-质谱（GC-MS）联用分析密蒙花（醉鱼草科）的挥发性花香成分

采用气相色谱-质谱联用技术对动态吸附法收集的密蒙花（Buddleja officinalis）花香成分进行了分析,并用气相色谱面积归-化法对各成分进行了定量。从密蒙花中分离出16个挥发性成分,定性定量出其中的11个,占挥发性成分总量的95．44％。其中丁基醋酸乙酯（81．57％）、苯甲醛（4．92％）、3-已烯-1-醇（3．26％）、欧洲丁香醛（2．34％）和芳樟醇（1．05％）为主要成分。该研究阐明了自然条件下密蒙花的花香成分及组成,其结果为今后定向创新醉鱼草属新香型观赏品种提供了科学依据。     

Effect of intravaginal instillation of some non-steroidal oral antifertility agents on pregnancy in rats

A study was conducted of the effect of intravaginal instillation of 3 nonsteroidal oran antifertility agents in pregnant rats. The 3 compounds used were 2-phenyl-3-rho-beta-pyrrolidinoethoxyphenyl-6-methox y benzofuran hydrochoride (DBF), 20 mg/kg; 2-phenyl-3-rho-beta-pyrrolidi noethoxyphenyl-naptha (2:1,b furan (NF), 10 mg/kg; and Centchroman, 1.25 mg/kg. These agents were administered on Day 1 postcoitum at the single-day oral contraceptive dose. Control animals were inserted with a pellet containing only carboxymethyl cellulose. Only Centchroman was 100% effective in preventing pregnancy. Even at half the normal dose, Centchroman was effective. Of the 3 compounds, only Centchroman has an antiprogestational property which may be contributing to Centchroman's high antifertility activity by the vaginal route and may prevent side effects.     

2-Ethoxybenzoxazole as a bioisosteric replacement of an ethyl benzoate group in a human rhinovirus (HRV) capsid binder

A series of pyridazinylpiperidinyl capsid-binding compounds with novel bicyclic substituents were synthesized and screened against human rhinovirus (HRV). Several 2-alkoxy- and 2-alkylthio-benzoxazole and benzothiazole derivatives showed excellent anti-HRV activity. When tested against a panel of 16 representative HRV types the 2-ethoxybenzoxazole derivative 13 was found to have superior HRV activity (median EC(50) 3.88ng/mL) to known capsid-binders Pleconaril and Pirodavir. Compound 13 illustrates that a 2-alkoxybenzoxazole group can be an effective bioisostere for a benzoate ester or benzaldehyde oxime ether functionality.     

Production of (S)-3-chlorolactaldehyde from (S)-alpha-chlorohydrin by boar spermatozoa and the inhibition of glyceraldehyde 3-phosphate dehydrogenase in vitro

The (S)-isomer of the male antifertility agent alpha-chlorohydrin was metabolized by mature boar spermatozoa in vitro to (S)-3-chlorolactaldehyde. This oxidative process, which did not occur when (R)-alpha-chlorohydrin was offered as a substrate, was catalysed by an NADP+-dependent dehydrogenase that converts glycerol to glyceraldehyde. (S)-3-chlorolactaldehyde, produced by this metabolic reaction or when added to suspensions of boar spermatozoa, was a specific inhibitor of glyceraldehyde 3-phosphate dehydrogenase as assessed by the accumulation of fructose 1,6-bisphosphate and the triosephosphates. When glycerol and (S)-alpha-chlorohydrin were added concomitantly to boar spermatozoa in vitro, the presence of glycerol decreased the degree of inhibition of glyceraldehyde 3-phosphate dehydrogenase. Extracts of glyceraldehyde 3-phosphate dehydrogenase that were obtained from boar spermatozoa incubated with (S)-alpha-chlorohydrin or (R,S)-3-chlorolactaldehyde showed significant reductions in their enzymic activity.     

Shifting the biotransformation pathways of L-phenylalanine into benzaldehyde by Trametes suaveolens CBS 334.85 using HP20 resin

AIMS: The biotransformation of L-phenylalanine into benzaldehyde (bitter almond aroma) was studied in the strain Trametes suaveolens CBS 334.85. METHODS AND RESULTS: Cultures of this fungus were carried out in the absence or in the presence of HP20 resin, a highly selective adsorbent for aromatic compounds. For the identification of the main catabolic pathways of L-phenylalanine, a control medium (without L-phenylalanine) was supplemented with each of the aromatic compounds, previously detected in the culture broth, as precursors. Trametes suaveolens CBS 334.85 was shown to biosynthesize benzyl and p-hydroxybenzyl derivatives, particularly benzaldehyde, and large amounts of 3-phenyl-1-propanol, benzyl and p-hydroxybenzyl alcohols as the products of both cinnamate and phenylpyruvate pathways. CONCLUSION: The addition of HP20 resin, made it possible to direct the catabolism of L- phenylalanine to benzaldehyde, the desired target compound, and to trap it before its transformation into benzyl alcohol. In these conditions, benzaldehyde production was increased 21-fold, from 33 to 710 mg l-1 corresponding to a molar yield of 31%. SIGNIFICANCE AND IMPACT OF THE STUDY: These results showed the good potential of Trametes suaveolens as a biotechnological agent to synthesize natural benzaldehyde which is one of the most important aromatic aldehydes used in the flavour industry.     

Synthesis and in vitro antimicrobial studies of some new 3-[phenyldiazenyl] benzaldehyde N-phenyl thiosemicarbazones

The increasing clinical importance of drug resistant microbial pathogens has lent additional urgency to microbiological research and new antimicrobial compound development. For this purpose, a new series of 3-[phenyldiazenyl] benzaldehyde N-phenylthiosemicarbazones were synthesized and evaluated for antifungal and antibacterial activity. The reaction of 2-hydroxy-5-[phenyldiazenyl] benzaldehyde (I) with N-phenylhydrazinecarbothioamide (II) were carried out in DMF. The antimicrobial activity of the synthesized target compounds (III) were evaluated by screening on different human pathogens using the disc diffusion assay. All the compounds exhibited considerable inhibition against the bacteria and fungi tested.     

Chemical sterilization of male langurs: synergistic action of alpha-chlorohydrin (U-5897) with methallibure (ICI, 33828) on the testes and epididymides of Presbytis entellus entellus Dufresne.

1. Synergistic action of alpha-chlorohydrin with methallibure (ICI, 33828) on the testicular function of Presbytis entellus entellus Dufresne has been studied. 2. Chronic administration of alpha-chlorohydrin alone (140 mg/day for 40 days) caused testicular lesion resulting in a massive atrophy of the spermatogenic elements. Epididymal epithelium was regressed and the lumen was devoid of spermatozoa. 3. alpha-Chlorohydrin inhibited the synthesis of RNA and sialic acid in the testes and epididymides. Total cholesterol per gram of testis and alkaline phosphatase activity were increased after alpha-chlorohydrin administration. 4. These effects could be achieved with a lower dose of alpha-chlorohydrin (1/4) when administered in combination with a gonadotrophin inhibitor, i. e. ICI, 33828 (Methallibure). Methallibure alone (200 mg/kg: total dose) has no damaging effects on the testes and epididymides. But it altered testicular cholesterol and enzyme activity. 5. In conclusion, an effective inhibition of spermatogenesis could be achieved by synergistic action of the two different drugs i. e. alpha-chlorohydrin and ICI, 33828 (Methallibure).     

Mode of action of alpha-chlorohydrin as a male anti-fertility agent. Inhibition of the metabolism of ram spermatozoa by alpha-chlorohydrin and location of block in glycolysis

1. The effect of alpha-chlorohydrin on the metabolism of glycolytic and tricarboxylate-cycle substrates by ram spermatozoa was investigated. The utilization and oxidation of fructose and triose phosphate were much more sensitive to inhibition by alpha-chlorohydrin (0.1-1.0mm) than lactate or pyruvate. Inhibition of glycolysis by alpha-chlorohydrin is concluded to be between triose phosphate and pyruvate formation. Oxidation of glycerol was not as severely inhibited as that of the triose phosphate. This unexpected finding can be explained in terms of competition between glycerol and alpha-chlorohydrin. A second, much less sensitive site, of alpha-chlorohydrin inhibition appears to be associated with production of acetyl-CoA from exogenous and endogenous fatty acids. 2. Measurement of the glycolytic intermediates after incubation of spermatozoal suspensions with 15mm-fructose in the presence of 3mm-alpha-chlorohydrin showed a ;block' in the conversion of glyceraldehyde 3-phosphate into 3-phosphoglycerate. alpha-Chlorohydrin also caused conversion of most of the ATP in spermatozoa into AMP. After incubation with 3mm-alpha-chlorohydrin, glyceraldehyde 3-phosphate dehydrogenase and triose phosphate isomerase activities were decreased by approx. 90% and 80% respectively, and in some experiments aldolase was also inhibited. Other glycolytic enzymes were not affected by a low concentration (0.3mm) of alpha-chlorohydrin. Loss of motility of spermatozoa paralleled the decrease in glyceraldehyde 3-phosphate dehydrogenase activity. alpha-Chlorohydrin, however, did not inhibit glyceraldehyde 3-phosphate dehydrogenase or triose phosphate isomerase in sonicated enzyme preparations when added to the assay cuvette. 3. Measurement of intermediates and glycolytic enzymes in ejaculated spermatozoa before, during and after injection of rams with alpha-chlorohydrin (25mg/kg body wt.) confirmed a severe block in glycolysis in vivo at the site of triose phosphate conversion into 3-phosphoglycerate within 24h of the first injection. Glyceraldehyde 3-phosphate dehydrogenase activity was no longer detectable and both aldolase and triose phosphate isomerase were severely inhibited. Spermatozoal ATP decreased by 92% at this time, being quantitatively converted into AMP. At 1 month after injection of alpha-chlorohydrin glycolytic intermediate concentrations returned to normal in the spermatozoa but ATP was still only 38% of the pre-injection concentration. Motility of spermatozoa was, however, as good as during the pre-injection period. The activity of the inhibited enzymes also returned to normal during the recovery period and 26 days after injection were close to pre-injection values. 4. An unknown metabolic product of alpha-chlorohydrin is suggested to inhibit glyceraldehyde 3-phosphate dehydrogenase and triose phosphate isomerase of spermatozoa. This results in a lower ATP content, motility and fertility of the spermatozoa. Glycidol was shown not to be an active intermediate of alpha-chlorohydrin in vitro.     

Cloning,expression, and genomic organization of mouse mp29 gene

The convulsions of approximately 25% of epileptics are inadequately controlled by currently available medication; therefore the preparation of new antiepileptic drugs is of great interest. Aryl semicarbazones can be considered a new class of compounds presenting anticonvulsant activity. In addition, they can be orally administered and are more active as anticonvulsants than mephenytoin or phenobarbital. However, one disadvantage of these compounds is their low water solubility. As a strategy to circumvent this problem, a 1:1 inclusion compound of benzaldehyde semicarbazone (BS) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was prepared and characterized. The anticonvulsant activities of the free semicarbazone and of the inclusion compound were evaluated in rats using the maximum electroshock and audiogenic seizures screenings. In both tests the minimum dose of compound necessary to produce activity decreases from 100mg/kg for the free semicarbazone to 35 mg/kg for the inclusion compound, indicating a significant increase in the bio-availability of the drug.     

Effects of alpha-chlorohydrin and gonadectomy on the adenohypophysial cells of male rats and gerbils

The cytological changes occurring in the gonadotropic cells of the anterior lobe of the pituitary in 30 male rats and 30 male gerbils after administration of alpha-chlorohydrin were compared with those occurring following bilateral gonadectomy. A decline in the weights of testes and accessory sex glands and inhibited spermatogenesis were noted in rats and gerbils following alpha-chlorohydrin administration. Alpha-cholorohydrin changed the appearance of the delta-basilphils, cell cytoplasm showed fine granulation and vacuolization, and castration vacuoles appeared in many delta-basiphils of the anteroir lobe of the pituitary. The compound brought about transient changes resembling those of castration in the anterior lobe of the pituitary of male rats and gerbils. In gerbils, there was also a marked decrease in the number of acidophils. The results show that alpha-chlorohydrin exerts its effects on androgen-dependent structures, e.g., seminal vesicles, ventral prostrate, epididymides, and perineal complex.     

Inhibition of the metabolism of ram spermatozoa by derivatives of α-chlorohydrin

The effects of the male antifertility agent, α-chlorohydrin, six of its derivatives, and glycidol were studied on the metabolism of washed ram spermatozoa in vitro with fructose as substrate. The α-chlorohydrin derivatives were the amino, the phosphorylated, and four glycol-bridge (ketal) compounds. All compounds except glycidol, in a concentration between 0.1 and 100 mM, reduced the aerobic glycolsis and/or oxidation of fructose. However, there was not a high correlation between the ability of these compounds to inhibit the metabolism of ram spermatozoa in vitro and their antifertility activity when administered to male rats. Other factors are clearly involved in their antifertility activity, eg, the concentration of the compounds in the epididymis and their conversion of either more or less spermicidal compounds in the body.